Laser-Induced Axotomy of Human iPSC-Derived and Murine Primary Neurons Decreases Somatic Tau and AT8 Tau Phosphorylation: A Single-Cell Approach to Study Effects of Acute Axonal Damage.

The microtubule-associated protein Tau is highly enriched in axons of brain neurons where it regulates axonal outgrowth, plasticity, and transport. Efficient axonal Tau sorting is critical since somatodendritic Tau missorting is a major hallmark of Alzheimer's disease and other tauopathies. However, the molecular mechanisms of axonal Tau sorting are still not fully understood. In this study, we aimed to unravel to which extent anterograde protein transport contributes to axonal Tau sorting. We developed a laser-based axotomy approach with single-cell resolution and combined it with spinning disk confocal microscopy enabling multi live-cell monitoring. We cultivated human iPSC-derived cortical neurons and mouse primary forebrain neurons in specialized chambers allowing reliable post-fixation identification and Tau analysis. Using this approach, we achieved high post-axotomy survival rates and observed axonal regrowth in a subset of neurons. When we assessed somatic missorting and phosphorylation levels of endogenous human or murine Tau at different time points after axotomy, we surprisingly did not observe somatic Tau accumulation or hyperphosphorylation, regardless of their regrowing activity, consistent for both models. These results indicate that impairment of anterograde transit of Tau protein and acute axonal damage may not play a role for the development of somatic Tau pathology. In sum, we developed a laser-based axotomy model suitable for studying the impact of different Tau sorting mechanisms in a highly controllable and reproducible setting, and we provide evidence that acute axon loss does not induce somatic Tau accumulation and AT8 Tau phosphorylation. UV laser-induced axotomy of human iPSC-derived and mouse primary neurons results in decreased somatic levels of endogenous Tau and AT8 Tau phosphorylation.

Axonal TAU Sorting Requires the C-terminus of TAU but is Independent of ANKG and TRIM46 Enrichment at the AIS.

Somatodendritic missorting of the axonal protein TAU is a hallmark of Alzheimer's disease and related tauopathies. Rodent primary neurons and iPSC-derived neurons are used for studying mechanisms of neuronal polarity, including TAU trafficking. However, these models are expensive, time-consuming, and/or require the killing of animals. In this study, we tested four differentiation procedures to generate mature neuron cultures from human SH-SY5Y neuroblastoma cells and assessed the TAU sorting capacity. We show that SH-SY5Y-derived neurons, differentiated with sequential RA/BDNF treatment, are suitable for investigating axonal TAU sorting. These human neurons show pronounced neuronal polarity, axodendritic outgrowth, expression of the neuronal maturation markers TAU and MAP2, and, importantly, efficient axonal sorting of endogenous and transfected human wild-type TAU, similar to mouse primary neurons. We demonstrate that the N-terminal half of TAU is not sufficient for axonal targeting, as a C-terminus-lacking construct (N-term-TAUHA) is not axonally enriched in both neuronal cell models. Importantly, SH-SY5Y-derived neurons do not show the formation of a classical axon initial segment (AIS), indicated by the lack of ankyrin G (ANKG) and tripartite motif-containing protein 46 (TRIM46) at the proximal axon, which suggests that successful axonal TAU sorting is independent of classical AIS formation. Taken together, our results provide evidence that (i) SH-SY5Y-derived neurons are a valuable human neuronal cell model for studying TAU sorting readily accessible at low cost and without animal need, and that (ii) efficient axonal TAU targeting is independent of ANKG or TRIM46 enrichment at the proximal axon in these neurons.

Erosion/abrasion-preventing potential of NaF and F/Sn/chitosan toothpastes in dentine and impact of the organic matrix.

The study investigated the erosion/abrasion-preventing potential of experimental NaF (1,400 ppm F(-)) and amine fluoride (AmF)/NaF/SnCl2/chitosan (1,400 ppm F(-), 3,500 ppm Sn(2+), 0.5% chitosan) toothpastes relative to placebo and SnF2 gel (970 ppm F(-), 3,000 ppm Sn(2+)), and the impact of the demineralised dentine matrix on toothpaste effects. The study was a cyclic erosion/intervention experiment (10 days). Samples were stored in mineral salt solution either without or with collagenase (from Clostridium histolyticum type VII; 100 U/ml) for continuous removal of the organic matrix. To produce a comparable order of tissue loss, erosion was performed 6 × 30 s/day with 0.5% citric acid in the latter and 6 × 90 s/day with 1% citric acid in the former. Intervention was toothpaste slurry immersion (2 × 2 min/day); half of the samples were additionally brushed for 15 s within this time (brushing machine, load 200 g). Tissue loss was determined profilometrically (mean ± SD; µm). Tissue loss values (without/with brushing) for placebo, NaF, AmF/NaF/SnCl2/chitosan and SnF2 gel, respectively, were 11.6 ± 3.1/12.2 ± 2.5, 12.7 ± 3.1/10.7 ± 4.5, 8.7 ± 2.1/9.7 ± 2.1 and 8.8 ± 1.8/​10.9 ± 1.8 in the presence of the organic matrix and 10.7 ± 3.2/11.9 ± 2.1, 8.2 ± 4.0/10.1 ± 4.1, 8.7 ± 2.9/9.1 ± 1.8 and 8.4 ± 1.9/7.5 ± 1.5 in the absence of the organic matrix. Relative to placebo, the NaF formulation had no significant effects; the AmF/NaF/SnCl2/chitosan formulation significantly reduced tissue loss between 20 and 25% except when applied without brushing in the absence of the organic matrix. The effects of the formulations were similar both in the presence and absence of the organic matrix. Sn(2+)/F(-)-​containing formulations have the potential to reduce erosion/abrasion even in the absence of demineralised collagen; seeking for more effective formulations is desirable.

Effect of a chitosan additive to a Sn2+-containing toothpaste on its anti-erosive/anti-abrasive efficacy--a controlled randomised in situ trial.

OBJECTIVES: It is well known that Sn(2+) is a notable anti-erosive agent. There are indications that biopolymers such as chitosan can enhance the effect of Sn(2+), at least in vitro. However, little information exists about their anti-erosive/anti-abrasive in situ effects. In the present in situ study, the efficacy of Sn(2+)-containing toothpastes in the presence or absence of chitosan was tested. METHODS: Ten subjects participated in the randomised crossover study, wearing mandibular appliances with human enamel specimens. Specimens were extraorally demineralised (7 days, 0.5% citric acid, pH 2.6; 6 × 2 min/day) and intraorally exposed to toothpaste suspensions (2 × 2 min/day). Within the suspension immersion time, one half of the specimens were additionally brushed intraorally with a powered toothbrush (5 s, 2.5 N). Tested preparations were a placebo toothpaste (negative control), two experimental toothpastes (F/Sn = 1,400 ppm F(-), 3,500 ppm Sn(2+); F/Sn/chitosan = 1,400 ppm F(-), 3,500 ppm Sn(2+), 0.5 % chitosan) and an SnF2-containing gel (positive control, GelKam = 3,000 ppm Sn(2+), 1,000 ppm F(-)). Substance loss was quantified profilometrically (µm). RESULTS: In the placebo group, tissue loss was 11.2 ± 4.6 (immersion in suspension) and 17.7 ± 4.7 (immersion in suspension + brushing). Immersion in each Sn(2+)-containing suspension significantly reduced tissue loss (p ≤ 0.01); after immersion in suspension + brushing, only the treatments with GelKam (5.4 ± 5.5) and with F/Sn/chitosan (9.6 ± 5.6) significantly reduced loss [both p ≤ 0.05 compared to placebo; F/Sn 12.8 ± 6.4 (not significant)] CONCLUSION: Chitosan enhanced the efficacy of the Sn(2+)-containing toothpaste as an anti-erosive/anti-abrasive agent. CLINICAL RELEVANCE: The use of Sn(2+)- and chitosan-containing toothpaste is a good option for symptomatic therapy in patients with regular acid impacts.

Randomised in situ study on the efficacy of a tin/chitosan toothpaste on erosive-abrasive enamel loss.

Tin is a notable anti-erosive agent, and the biopolymer chitosan has also shown demineralisation-inhibiting properties. Therefore, the anti-erosive/anti-abrasive efficacy of the combination of both compounds was tested under in situ conditions. Twenty-seven volunteers were included in a randomised, double-blind, three-cell crossover in situ trial. Enamel specimens were recessed on the buccal aspects of mandibular appliances, extraorally demineralised (6 × 2 min/day) and intraorally treated with toothpaste slurries (2 × 2 min/day). Within the slurry treatment time, one-half of the specimens received additional intraoral brushing (5 s, 2.5 N). The tested toothpastes included a placebo toothpaste, an experimental NaF toothpaste (1,400 ppm F(-)) and an experimental F/Sn/chitosan toothpaste (1,400 ppm F(-), 3,500 ppm Sn(2+), 0.5% chitosan). The percentage reduction of tissue loss (slurry exposure/slurry exposure + brushing) compared to placebo was 19.0 ± 47.3/21.3 ± 22.4 after use of NaF and 52.5 ± 30.9/50.2 ± 34.3 after use of F/Sn/chitosan. F/Sn/chitosan was significantly more effective than NaF (p ≤ 0.001) and showed good efficacy against erosive and erosive-abrasive tissue loss. This study suggests that the F/Sn/chitosan toothpaste could provide good protection for patients who frequently consume acidic foodstuffs.

Efficacy of a moisture-tolerant material for fissure sealing: a prospective randomised clinical trial.

OBJECTIVES: Fissure sealings offer nearly complete protection against fissure caries, provided that they are adequately applied, for composite-based sealants with sufficient moisture control. This is not always attainable, particularly in children with low compliance. To counter this problem, a moisture-tolerant sealant has been developed. The present randomised clinical trial compared such a moisture-tolerant material (Embrace) with a conventional sealant (Helioseal). MATERIAL AND METHODS: In 55 participants (mean age, 10 ± 3 years), corresponding molar pairs were sealed with either Embrace or Helioseal. Retention, quality of sealing, and caries were clinically examined, both tactilely and visually, immediately and after 1 year. RESULTS: After 1 year, 93% of Helioseal sealings were complete, whereas 60% of Embrace sealings showed partial and 13% complete loss. The surface quality of Embrace was significantly worse than that of Helioseal. After the use of Embrace, the sealant margin was noticeable as a slight (distinct) step in 36% (15%). The visual (tactile) examination showed a rough surface in 78% (33%) in the case of Embrace. The Helioseal surfaces were shiny (smooth) in all cases (all differences between Helioseal and Embrace, p ≤ 0.001). Caries was found only after the use of Embrace (4%, n.s. compared to Helioseal). CONCLUSION: The moisture-tolerant material Embrace was distinctly inferior to Helioseal because Embrace showed weaknesses in retention and surface quality. CLINICAL RELEVANCE: Even if a moisture-tolerant sealant would be desirable in particular for children with low compliance, the tested material does not represent an alternative to the standard preparation.

Relationship between plaque score and video-monitored brushing performance after repeated instruction--a controlled, randomised clinical trial.

OBJECTIVES: Aim of this prospective, randomised, controlled clinical trial was to use the modified bass technique (MBT) and a specific brushing sequence to investigate whether two types of instruction methods lead to differences in plaque reduction and whether plaque reduction is related to technique adoption. METHODS: Ninety-eight participants were randomly assigned to three groups: (1) control, no instruction; (2) verbal instruction by means of a leaflet; and (3) verbal instruction supported by demonstration, no leaflet. Brushing performance was video monitored. Plaque score (Turesky modified QHI (T-QHI)) was measured at baseline, afterwards participants received instructions. After 2 weeks, T-QHI was measured for a second time, and participants were re-instructed. After another 2 weeks, T-QHI was measured for a third time. RESULTS: At baseline, T-QHI did not differ between groups ((1) 1.99 ± 0.51, (2) 1.90 ± 0.51, (3) 1.93 ± 0.56). The second measurement revealed an improvement of T-QHI in the instructed groups and in the non-instructed control group ((1) 1.80 ± 0.47, (2) 1.58 ± 0.58, (3) 1.64 ± 0.58; n.s. between groups); in the intervention groups, remotivation achieved no further improvement ((1) 1.72 ± 0.48, (2) 1.52 ± 0.58, (3) 1.50 ± 0.69; n.s. between groups and compared to second measurement). Improvement of T-QHI was not related to proper performance of technique or brushing sequence. Those who fully adopted the brushing technique, the sequence or both did not have lower plaque scores. CONCLUSION: Technical performance and effectiveness were not linked. CLINICAL RELEVANCE: Within the study setting, the MBT was not effective in reducing plaque scores. The general recommendation of the MBT should be re-evaluated in further studies.

Efficacy of the stannous ion and a biopolymer in toothpastes on enamel erosion/abrasion.

OBJECTIVES: Sn(2+) has promising erosion-inhibiting properties in solutions, but little is known about respective effects in toothpastes. In addition, biopolymers might have protecting potential. Aim of this study was to investigate the effects of Sn(2+) in toothpastes and of a biopolymer (chitosan) added to a Sn(2+) formulation on erosion/abrasion. METHODS: Enamel samples were subjected to cyclic erosion procedures (10 days; 0.50% citric acid, pH 2.5; 6× 2 min/day), and brushing (2× 15s/day, load 200 g) during immersion in slurries (2 min). The toothpastes were NaF formulations (NaF/1, NaF/2, NaF/3) and Sn(2+) formulations (NaF/SnCl(2), AmF/SnF(2), AmF/NaF/SnCl(2)) and AmF/NaF/SnCl(2)+0.5% chitosan. Declared concentrations of active ingredients in toothpastes were 1400-1450 µg/g F(-) and 3280-3500 µg/g Sn(2+). Negative controls were erosion only and placebo, positive control was a SnF(2) gel. Tissue loss was quantified profilometrically, Sn on enamel surfaces was measured by energy dispersive X-ray spectroscopy. RESULTS: Loss values (µm) for erosion only and placebo were 14.4 ± 4.5 and 20.2 ± 3.8, respectively, and 4.6 ± 1.9 for the positive control (p ≤ 0.001 each compared to erosion only). The other loss values were: NaF/1 16.5 ± 3.0, NaF/2 14.0 ± 2.7, NaF/3 12.6 ± 3.9, NaF/SnCl(2) 14.7 ± 5.1, AmF/SnF(2) 13.5 ± 4.8, AmF/NaF/SnCl(2) 12.4 ± 4.2, AmF/NaF/SnCl(2)+chitosan 6.6 ± 3.5 (except NaF/1 all p ≤ 0.01 compared to placebo). AmF/NaF/SnCl(2)/chitosan was more effective than all other toothpastes (p ≤ 0.01 each). Sn on the enamel surface ranged between 1.3 ± 0.3 and 2.8 ± 0.04 wt.% with no obvious relationship with efficacy. CONCLUSIONS: The NaF and Sn(2+) toothpastes without chitosan exhibited similar anti-erosion and abrasion-prevention effects. The experimental Sn(2+) formulation with chitosan revealed promising results similar to those of the positive control. CLINICAL SIGNIFICANCE: NaF toothpastes offer a degree of protection against erosion/abrasion, which is likely sufficient for most subjects with average acid exposures. For patients with initial erosive lesions, however, more effective toothpaste is desirable. The combination of Sn(2+) and a biopolymer appears promising in this context.

Enzyme activities in the oral fluids of patients suffering from bulimia: a controlled clinical trial.

Patients with bulimia nervosa are at high risk for dental erosion. However, not all bulimic patients suffer from erosion, irrespective of the severity of their eating disorder. It is often speculated that differences in the saliva are important, however, little is known about salivary parameters in bulimic patients, particularly directly after vomiting. The aim of the clinical trial was to compare different salivary parameters of subjects suffering from bulimia with those of healthy controls. Twenty-eight subjects participated (14 patients with bulimia nervosa, 7 of them with erosion; 14 matched healthy controls). Resting and stimulated saliva of all participants was analysed as well as saliva collected from bulimic patients directly and 30 min after vomiting. Parameters under investigation were flow rate, pH, buffering capacity and the enzyme activities of proteases in general, collagenase, pepsin, trypsin, amylase, peroxidase, and lysozyme. Regarding flow rate, pH and buffering capacity only small differences were found between groups; buffering capacity directly after vomiting was significantly lower in bulimic subjects with erosion than in subjects without erosion. Differences in enzymatic activities were more pronounced. Activities of proteases, collagenase and pepsin in resting and proteases in stimulated saliva were significantly higher in bulimic participants with erosion than in controls. Peroxidase activity was significantly decreased by regular vomiting. Proteolytic enzymes seem to be relevant for the initiation and progression of dental erosion directly after vomiting, maybe by both hydrolysis of demineralized dentine structures as well as modulation of the pellicle layer.

Conventional and anti-erosion fluoride toothpastes: effect on enamel erosion and erosion-abrasion.

New toothpastes with anti-erosion claims are marketed, but little is known about their effectiveness. This study investigates these products in comparison with various conventional NaF toothpastes and tin-containing products with respect to their erosion protection/abrasion prevention properties. In experiment 1, samples were demineralised (10 days, 6 × 2 min/day; citric acid, pH 2.4), exposed to toothpaste slurries (2 × 2 min/day) and intermittently stored in a mineral salt solution. In experiment 2, samples were additionally brushed for 15 s during the slurry immersion time. Study products were 8 conventional NaF toothpastes (1,400-1,490 ppm F), 4 formulations with anti-erosion claims (2 F toothpastes: NaF + KNO(3) and NaF + hydroxyapatite; and 2 F-free toothpastes: zinc-carbonate-hydroxyapatite, and chitosan) and 2 Sn-containing products (toothpaste: 3,436 ppm Sn, 1,450 ppm F as SnF(2)/NaF; gel: 970 ppm F, 3,030 ppm Sn as SnF(2)). A mouth rinse (500 ppm F as AmF/NaF, 800 ppm Sn as SnCl(2)) was the positive control. Tissue loss was quantified profilometrically. In experiment 1, most NaF toothpastes and 1 F-free formulation reduced tissue loss significantly (between 19 and 42%); the Sn-containing formulations were the most effective (toothpaste and gel 55 and 78% reduction, respectively). In experiment 2, only 4 NaF toothpastes revealed significant effects compared to the F-free control (reduction between 29 and 37%); the F-free special preparations and the Sn toothpaste had no significant effect. The Sn gel (reduction 75%) revealed the best result. Conventional NaF toothpastes reduced the erosive tissue loss, but had limited efficacy regarding the prevention of brushing abrasion. The special formulations were not superior, or were even less effective.

AMP-deaminase from human preterm placenta--kinetic regulatory properties of enzyme.

During pregnancy the isoform composition of human placental AMP-deaminase changes. This may reflect the adaptation of enzyme to changing metabolic requirements of the growing fetus. In this paper kinetic and regulatory properties of AMP-deaminase purified from human preterm (∼ 25 week of gestation) placenta were described and compared with these of the enzyme purified from term placenta. AMP-deaminase from preterm placenta was less sensitive to pH changes and in contrast to the enzyme from the term organ, at low range of substrate concentrations was not inhibited but activated by physiological concentrations of orthophosphate. This may significantly improve the catalytic efficiency of enzyme at early phase of the pregnancy.

Ventilator-delivered mask ventilation compared with three standard methods of mask ventilation in a manikin model.

BACKGROUND: Little is known regarding the variations in effective ventilation during bag and mask resuscitation with standard methods compared with that delivered by ventilator-delivered mask ventilation (VDMV). AIM: To measure the variations in delivered airway pressure, tidal volume (TV), minute ventilation (MV) and inspiratory time during a 3-min period of mask ventilation comparing VDMV with three commonly used hand-delivered methods of bag and mask ventilation: Laerdal self-inflating bag (SIB); anaesthetic bag and T-piece Neopuff. METHODS: A modified resuscitation manikin was used to measure variation in mechanical ventilation during 3-min periods of mask ventilation. Thirty-six experienced practitioners gave positive pressure mask ventilation targeting acceptable chest wall movement with a rate of 60 inflations/min and when pressures could be targeted or set, a peak inspiratory pressure (PIP) of 18 cm water, positive end-expiratory pressure (PEEP) of 5 cm water, for 3 min with each of the four mask ventilation methods. Each mode was randomly sequenced. RESULTS: A total of 21 136 inflations were recorded and analysed. VDMV achieved PIP and PEEP closest to that targeted and significantly lower variation in all measured parameters (p<0.001) other than with PIP. SIB delivered TV and MV over twice that delivered by VDMV and Neopuff. CONCLUSION: During 3-min periods of mask ventilation on a manikin, VDMV produced the least variation in delivered ventilation. SIB produced wide variation and unacceptably high TV and MV in experienced hands.

Mask leak in one-person mask ventilation compared to two-person in newborn infant manikin study.

AIM: To compare a new two-person method (four hands) of delivering mask ventilation with a standard one-person method using the Laerdal self-inflating bag (SIB) and the Neopuff (NP) infant resuscitator in a manikin model. BACKGROUND: Recent studies of simulated neonatal resuscitation using bag and mask ventilation techniques have shown facemask leak levels of 55-57% in expert hands. METHODS: 48 participants were randomly paired and instructed to give mask ventilation for a 2-min period as single-person resuscitators, then as two-person paired resuscitators at set pressures for NP and set parameters for SIB. Airway pressure, flow, inspiratory tidal volume, expiratory tidal volume and mask leak were recorded. RESULTS: A total of 21 578 inflations were recorded and analysed. For SIB, mask leak was greater (11.5%) with single-person compared to two-person (5.4%; mean difference 6.1%, 95% CI 1.5 to 10.7, p<0.01). For NP, mask leak was greater for single-person (22.2%) compared to two-person (9.1%; mean difference 13.1% 95% CI 3.6 to 22.6, p<0.01). For single-person mask ventilation, mask leak was greater with NP (22.2%) compared to SIB (11.5%; mean difference 10.7%, 95% CI 1.4 to 19.7, p<0.01). For two-person mask ventilation, mask leak was greater for NP (9.1%) compared to SIB (5.4%; mean difference 3.7%, 95% CI 0.1 to 6.4, p<0.05). CONCLUSIONS: Two-person mask ventilation technique reduces mask leak by approximately 50% compared to the standard one-person mask ventilation method. NP mask ventilation has higher mask leak than Laerdal SIB for both single- and two-person technique mask ventilation.

Efficacy of a tin/fluoride rinse: a randomized in situ trial on erosion.

Concentrated tin- and fluoride-containing mouthrinses are effective erosion inhibitors in enamel and dentin. To test whether this is also true for solutions with lower concentrations, we conducted a randomized double-blind three-cell crossover in situ study with extra-orally performed erosive impacts (citric acid, 6 x 5 min/day) and an intra-oral rinsing protocol (1 x 30 sec/day) in 24 volunteers. The mouthrinses were a placebo, a NaF (500 ppm F⁻), and an amine fluoride (AmF)/NaF/SnCl2 mouthrinse (500 ppm F⁻, 800 ppm Sn(2+)). Compared with the placebo, the NaF mouthrinse reduced substance loss by 19% in enamel and 23% in dentin (p ≤ 0.01 each); the AmF/NaF/SnCl2 mouthrinse reduced this parameter by 67% in enamel and 47% in dentin (p ≤ 0.001 each). AmF/NaF/SnCl2 was significantly more effective than NaF in both tissues (p ≤ 0.01). The mouthrinse containing Sn and F exhibited good efficacy, even under severe erosive conditions.

Cardiac muscle AMP-deaminase from a 10-year-old male heterozygous for the AMPD1 C34T mutation.

A C34T mutation in the AMPD1 gene is proposed to cause local or systemic augmentations in blood adenosine level and improvement of prognoses in heart diseases like congestive heart failure or heart ischemic disease. This study examines some physico-chemical properties of AMP-deaminase isolated from cardiac muscle of a 10-year-old boy heterozygote for this mutation.

Efficacy of fluoride compounds and stannous chloride as erosion inhibitors in dentine.

The aim of this study was to evaluate the anti-erosive effects of different fluoride compounds and one tin compound in the context of the complex pathohistology of dentine erosion, with particular emphasis on the role of the organic portion. Samples were subjected to two experiments including erosive acid attacks (0.05 molar citric acid, pH 2.3; 6 x 2 min/day) and applications (6 x 2 min/day) of the following test solutions: SnCl(2) (815 ppm Sn), NaF (250 ppm F), SnF(2) (250 ppm F, 809 ppm Sn), amine fluoride (AmF, 250 ppm F), AmF/NaF (250 ppm F), and AmF/SnF(2) (250 ppm F, 409 ppm Sn). The demineralised organic fraction was enzymatically removed either at the end of the experiment (experiment 1) or continuously throughout the experiment (experiment 2). Tissue loss was determined profilometrically after 10 experimental days. In experiment 1, the highest erosive tissue loss was found in the control group (erosion only); the AmF- and NaF-containing solutions reduced tissue loss by about 60%, reductions for SnCl(2), AmF/SnF(2), and SnF(2) were 52, 74 and 89%, respectively. In experiment 2, loss values generally were significantly higher, and the differences between the test solutions were much more distinct. Reduction of tissue loss was between 12 and 34% for the AmF- and NaF-containing preparations, and 11, 67 and 78% for SnCl(2), AmF/SnF(2), and SnF(2), respectively. Stannous fluoride-containing solutions revealed promising anti-erosive effects in dentine. The strikingly different outcomes in the two experiments suggest reconsidering current methodologies for investigating anti-erosive strategies in dentine.

Neopuff T-piece mask resuscitator: is mask leak related to watching the pressure dial?

AIM: The aim of the study is to compare mask leak and delivered ventilation during Neopuff (NP) mask ventilation in two modes: (i) with NP pressure dial hidden and resuscitator watching chest wall (CW) rise with, (ii) CW movement hidden and resuscitator watching NP pressure dial. METHODS: Thirty-six participants gave mask ventilation to a modified manikin designed to measure mask leak and delivered ventilation for two minutes in each mode randomly assigned. Paired t-tests were used to analyse differences in mean values. Linear regression was used to determine the association of mask leak with delivered ventilation. RESULTS: Of 7277 inflations analysed, 3621 were observing chest wall mode (CWM) and 3656 observing NP mode (NPM). Mask leak was similar between the groups; 31.6% for CWM and 31.5% (p = 0.56) for NPM. There were no significant differences in airways pressures and expired tidal volumes (TVe) between modes. Mask leak was strongly associated with TVe (R = -0.86 p < 0.0001) and with peak inspiratory pressure (PIP) (R = -0.51 p < 0.0001). TVe was associated with PIP (R = 0.51 p < 0.0001). CONCLUSION: This study provides reassurance that NP mask leak is not greater when resuscitators watch the NP pressure dial. Mask leak is related to TVe. Mask ventilation training with manikins should include tidal volume measurements.

Does caffeine impair cerebral oxygenation and blood flow velocity in preterm infants?

AIM: The aim of the study is to assess the effects of an intravenous 10 mg/kg loading dose of caffeine base in cerebral oxygenation, cerebral Doppler blood flow velocity and cardiac output in preterm infants. METHODS: Preterm neonates <34 weeks gestation were investigated at 1 and 4 h following the loading dose of caffeine using Doppler cerebral sonography, cardiac echocardiography and cerebral spatially resolved near-infrared spectroscopy. RESULTS: Forty infants were studied with a mean gestational age (mean ± standard deviation) of 27.7 (±2.5) weeks, birth weight of 1155 (±431) g and a postnatal age of 2.8 (±2.2) days. Mean Anterior Cerebral Artery peak and time average mean blood flow velocity fell significantly by 14% and 17.7%, respectively at 1 h post-caffeine loading dose, which recovered partially by 4 h. Cerebral Tissue Oxygenation Index fell from pre-dose levels by 9.5% at 1 h with partial recovery to 4.9% reduced at 4 h post-dose. There were no significant changes in left or right ventricular output, transcutaneous oxygen saturation, transcutaneous PCO(2) or total vascular resistance. CONCLUSIONS: A loading dose of 10 mg/kg caffeine base resulted in significant reduction at 1 h post-dose in cerebral oxygenation and cerebral blood flow velocity with partial recovery at 4 h.

AMP-deaminase from developing human placenta.

During pregnancy the activity of AMP-deaminase in developing human placenta gradually decreases, being in homogenates of mature, term placenta (approximately 40 week of gestation) one fourth to one third of that in homogenates of immature (approximately 25 week of gestation) organ. The gradual decrease of activity correlates inversely with the increasing presence of the form of enzyme predominating in homogenates of the mature placenta. The discrepancy observed indicate that isozymic pattern of AMP-deaminase in developing human placenta changes.

Adoption of a toothbrushing technique: a controlled, randomised clinical trial.

The aim of this study was to evaluate the implementation of the modified Bass technique (MBT) and a brushing sequence using different instruction methods. Ninety-nine participants, aged 19-42, were randomly assigned to one of three groups (control group: no instruction; leaflet instruction group: verbal instruction using a leaflet; and demonstration group: verbal instruction supported by demonstration with a model, no leaflet). Participants were instructed twice with an interval of 2 weeks. To evaluate the implementation of the technique and brushing sequence, participants were filmed during toothbrushing at baseline and 2 weeks after the first and second instruction, respectively. The duration of brushing was measured. After the first instruction, 19% in the leaflet instruction group and 41% in the demonstration group fully performed the MBT, and 36% in both instruction groups fully adopted the brushing sequence. After the second instruction, 25% of patients in the leaflet instruction group and 62% in the demonstration group had adopted the technique completely. The brushing sequence was adopted by 63% in the leaflet instruction group and by 48% in the demonstration group. Only 16% in the leaflet group and 38% in the demonstration group adopted both the technique and brushing sequence after the second instruction. The results indicate the need to improve instructional strategies.