In Vitro Microleakage of Composite Restorations Prepared by Er:YAG/Er,Cr:YSGG Lasers and Conventional Drills Associated with Two Adhesive Systems

Purpose: The objective of this in vitro study was to compare the degree of microleakage of composite restorations performed by lasers and conventional drills associated with two adhesive systems. Materials and Methods: Sixty bovine teeth were divided into 6 groups (n = 10). The preparations were performed in groups 1 and 2 with a high-speed drill (HID), in groups 3 and 5 with Er:YAG laser, and in groups 4 and 6 with Er,Cr:YSGG laser. The specimens were restored with resin composite associated with an etch-and-rinse two-step adhesive system (Single Bond 2 [SB]) (groups 1, 3, 4) and a self-etching adhesive (One-Up Bond F [OB]) (groups 2, 5, 6). After storage, the specimens were polished, thermocycled, immersed in 50% silver nitrate tracer solution, and then sectioned longitudinally. The specimens were placed under a stereomicroscope (25X) and digital images were obtained. These were evaluated by three blinded evaluators who assigned a microleakage score (0 to 3). The original data were submitted to Kruskal-Wallis and Mann-Whitney statistical tests. Results: The occlusal/enamel margins demonstrated no differences in microleakage for all treatments (p > 0.05). The gingival/dentin margins presented similar microleakage in cavities prepared with Er:YAG, Er,Cr:YSGG, and HD using the etch-and-rinse two-step adhesive system (SB) (p > 0.05); otherwise, both Er:YAG and Er,Cr:YSGG lasers demonstrated lower microleakage scores with OB than SB adhesive (p < 0.05). Conclusion: The microleakage score at gingival margins is dependent on the interaction of the hard tissue removal tool and the adhesive system used. The self-etching adhesive system had a lower microleakage score at dentin margins for cavities prepared with Er:YAG and Er,Cr:YSGG than the etch-and-rinse two-step adhesive system.

Influence of the bonding substrate in dental composite polymerization stress testing

Our objective was to compare the polymerization stress (sigma(pol)) of a series of composites obtained using poly(methyl methacrylate) (PMMA) or glass as bonding substrates, and to compare the results with those from in vitro microleakage of composite restorations. The tested hypothesis was that stress values obtained in a less rigid testing system (i.e. using PMMA) would show a better relationship with microleakage data. Five dental composites were tested: Filtek Z250 (FZ), Z100 (Z1), Concept (CO), Durafill (DU) and Heliomolar (HM). sigma(pol) was determined in 1 mm high specimens inserted between two rods (empty set = 5 mm) of either PMMA or glass. The composite elastic modulus (E) was obtained by three-point bending. sigma(pol) and E data were submitted to a one-way analysis of variance/Tukey test (alpha = 0.05). For the microleakage test (MI), bovine incisors received cylindrical cavities (empty set = 5 mm, h = 2 mm), which were restored in bulk. After storage for 24 h in water, specimens were subjected to dye penetration using AgNO(3) as tracer. Specimens were sectioned twice, perpendicularly, and microleakage was measured (in millimeters) under 20x magnification. Data from MI were submitted to the Kruskal-Wallis test. Means (SD) of sigma(pol) (MPa) using glass/PMMA were FZ: 7.5(1.8)(A)/2.5(0.2)(bc); Z1: 7.3(0.5)(A)/2.8(0.3)(ab); CO: 6.8(1.1)(A)/3.2(0.5)(a); DU: 4.5(0.7)(B)/2.0(0.2)(bc); HM: 3.5(0.2)(B)/2.3(0.3)(c). sigma(pol) obtained using PMMA rods were 34-67% lower than with glass. Means (SD) for tooth average/tooth maximum microleakage were FZ: 0.92(0.19)(B)/1.53(0.30)(a); Z1: 1.19(0.21)(A)/1.75(0.20)(a); CO: 1.26(0.25)(A)/1.78(0.24)(a); DU: 0.83(0.30)(B)/1.68(0.46)(a): HM: 0.81(0.27)(B)/1.64(0.54)(a). The tested hypothesis was confirmed, as the composites showed the same ordering both in the polymerization stress test using PMMA rods and in the microleakage test. (C) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Nanoscale Oxidative Patterning of Metallic Surfaces to Modulate Cell Activity and Fate

In the field of regenerative medicine, nanoscale physical cuing is clearly becoming a compelling determinant of cell behavior. Developing effective methods for making nanostructured surfaces with well-defined physicochemical properties is thus mandatory for the rational design of functional biomaterials. Here, we demonstrate the versatility of simple chemical oxidative patterning to create unique nanotopographical surfaces that influence the behavior of various cell types, modulate the expression of key determinants of cell activity, and offer the potential of harnessing the power of stem cells. These findings promise to lead to a new generation of improved metal implants with intelligent surfaces that can control biological response at the site of healing.

A comparison of anterior adhesive areas and secretions in Troglocephalus rhinobatidis and Neoheterocotyle rhinobatidis (Monogenea : Monocotylidae) from the gills of the shovelnose ray, Rhinobatos typus (Rhinobatidae)

This study continues the collection of data on the anterior adhesive areas and secretions of monopisthocotylean monogenean (flatworm) parasites and begins an investigation of their phylogenetic usefulness. Here, two species of parasitic worms from an elasmobranch, Troglocephalus rhinobatidis (Monocotylidae: Dasybatotreminae) and Neoheterocotyle rhinobatidis (Monocotylidae: Heterocotylinae), are compared and contrasted. It has been suggested in recent literature that these two taxa are more closely related than is currently recognised. Our data support this view. Both species have multiple apertures on the ventral anterior margin through which adhesive is secreted. Two types of secretion exit from multiple adjacent duct endings terminating in each aperture: rod-shaped (S1) and spherical-shaped (S2) bodies. S1 bodies of both species show nano-banding of similar size and are membrane bound. Ultrastructure of the glands, ducts, duct endings and secreted adhesive is similar for both species, but aperture shape differs. Away from the adhesive areas, tegumental inclusions are found to differ between the two species and another, apparently non-adhesive, secretion is found in N. rhinobatidis.

Projections from the substantia nigra pars reticulata to the motor thalamus of the rat: Single axon reconstructions and immunohistochemical study

This is a study in the rat of the distribution of specific neurotransmitters in neurones projecting from the substantia nigra reticulata (SNR) to the ventrolateral (VL) and ventromedial (VM) thalamic nuclei. Individual axons projecting from the SNR to these thalamic nuclei have also been reconstructed following small injection of the anterograde tracer dextran biotin into the the SNR. Analysis of reconstructions revealed two populations of SNR neurones projecting onto the VL and VM thalamic nuclei. One group projects directly onto the VM and VL, and the other projects to the VM/VL and to the parafascicular nucleus. In another set of experiments Fluoro-Gold was injected into the VL/VM to label SNR projection neurones retrogradely, and immunohistochemistry was performed to determine the distribution of choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), gamma -aminobutyric acid (GABA), and glutamate in Fluoro-Gold-labelled SNR projection neurones. Most SNR-VL/VM thalamic projection neurones were immunoreactive to acetylcholine or glutamate, whereas only 25% of the projection neurones were found to be immunoreactive to GABA. (C) 2001 Wiley-Liss, Inc.

Medullary neurones regulate hypothalamic corticotropin-releasing factor cell responses to an emotional stressor

Hypothalamic-pituitary-adrenal axis activation is a hallmark of the stress response. In the case of physical stressors, there is considerable evidence that medullary catecholamine neurones are critical to the activation of the paraventricular nucleus corticotropin-releasing factor cells that constitute the apex of the hypothalamic-pituitary-adrenal axis. In contrast, it has been thought that hypothalamic-pituitary-adrenal axis responses to emotional stressors do not involve brainstem neurones. To investigate this issue we have mapped patterns of restraint-induced neuronal c fos expression in intact animals and in animals prepared with either paraventricular nucleus-directed injections of a retrograde tracer, lesions of paraventricular nucleus catecholamine terminals, or lesions of the medulla corresponding to the A1 or A2 noradrenergic cell groups. Restraint-induced patterns of neuronal activation within the medulla of intact animals were very similar to those previously reported in response to physical stressors, including the fact that most stressor-responsive, paraventricular nucleus-projecting cells were certainly catecholaminergic and probably noradrenergic. Despite this, the destruction of paraventricular nucleus catecholamine terminals with 6-hydroxydopamine did not alter corticotropin-releasing factor cell responses to restraint. However, animals with ibotenic acid lesions encompassing either the A1 or A2 noradrenergic cell groups displayed significantly suppressed corticotropin-releasing factor cell responses to restraint. Notably, these medullary lesions also suppressed neuronal responses in the medial amygdala, an area that is now considered critical to hypothalamic-pituitary-adrenal axis responses to emotional stressors and that is also known to display a significant increase in noradrenaline turnover during restraint. We conclude that medullary neurones influence corticotropin-releasing factor cell responses to emotional stressors via a multisynaptic pathway that may involve a noradrenergic input to the medial amygdala. These results overturn the idea that hypothalamic-pituitary-adrenal axis response to emotional stressors can occur independently of the brainstem. (C) 2001 IBRO. Published by Elsevier Science Ltd. All rights reserved.

Combined hydraulic and biological modelling and full-scale validation of SBR process

The biological reactions during the settling and decant periods of Sequencing Batch Reactors (SBRs) are generally ignored as they are not easily measured or described by modelling approaches. However, important processes are taking place, and in particular when the influent is fed into the bottom of the reactor at the same time (one of the main features of the UniFed process), the inclusion of these stages is crucial for accurate process predictions. Due to the vertical stratification of both liquid and solid components, a one-dimensional hydraulic model is combined with a modified ASM2d biological model to allow the prediction of settling velocity, sludge concentration, soluble components and biological processes during the non-mixed periods of the SBR. The model is calibrated on a full-scale UniFed SBR system with tracer breakthrough tests, depth profiles of particulate and soluble compounds and measurements of the key components during the mixed aerobic period. This model is then validated against results from an independent experimental period with considerably different operating parameters. In both cases, the model is able to accurately predict the stratification and most of the biological reactions occurring in the sludge blanket and the supernatant during the non-mixed periods. Together with a correct description of the mixed aerobic period, a good prediction of the overall SBR performance can be achieved.

Axial mixing in rotating drums using magnetic resonance imaging using bran as a model for solid state fermentations

Magnetic resonance imaging (MRI) is an easily automated, reliable technique to investigate axial mixing within rotating drums. Moist bran can be clearly differentiated from dry bran using MRI allowing a non-segregating tracer for axial mixing. For a 20-cm diameter drum, the axial dispersion coefficient in the particle bed was 0.51 cm s(-2). Axial dispersion is scale-dependent.

Fate of N-15-labelled nitrate in a wet summer under different residue management regimes in young hoop pine plantations

A field study was conducted to investigate the fate of N-15-labelled nitrate applied at 20 kg N ha(-1) in a wet summer to microplots installed in areas under different residue management regimes in second-rotation hoop pine (Araucaria cunninghamii) plantations aged 1-3 years in south-east Queensland, Australia. PVC microplots of 235 mm diameter and 300 mm long were driven into 250 mm soil. There were three replications of each of eight treatments. These were areas just under and between 1-year-old windrows (ca. 2-3 m in width) of harvesting residues spaced 15 m apart, and with and without incorporated foliage residues (20 t DM ha(-1)); the areas just under and between 2- or 3-year-old windrows spaced 10 m apart. Only 7-29% of the added N-15 was recovered from the top 750 mm of the soil profile with the leaching loss estimated to be 70-86% over the 34-day period. The N-15 loss via denitrification was 3.7-6.3% by directly measuring the N-15 gases emitted. The microplots with the incorporated residues at the 1-year-old site had the highest N-15 loss (6.3%) as compared with the other treatments. The N-15 mass balance method together with the use of bromide (Br) tracer applied at 100 kg Br ha(-1) failed to obtain a reliable estimate of the denitrification loss. The microplots at the 1-year-old site had higher N-15 immobilisation rate (7.5-24.7%) compared with those at 2- and 3-year-old sites (2.1-3.6%). Incorporating the residues resulted in an increase in N-15 immobilisation rate (24.5-24.7%) compared with the control without the incorporated residues (8.4-14.3%). These findings suggest that climatic conditions played important roles in controlling the N-15 transformations in the wet summer season and that the residue management regimes could also significantly influence the N-15 transformations. Most of the N-15 loss occurred through leaching, but a considerable amount of the N-15 was lost through denitrification. Bromide proved to be an unsuitable tracer for monitoring the N-15 leaching and movement under the wet summer conditions. (C) 2002 Elsevier Science B.V. All rights reserved.

Systemic administration of interleukin-1 beta activates select populations of central amygdala afferents

The central nucleus of the amygdala (CeA) is activated robustly by an immune challenge such as the systemic administration of the proinflammatory cytokine interleukin-1beta (IL-1beta). Because IL-1beta is not believed to cross the blood-brain barrier in any significant amount, it is likely that IL-1beta elicits CeA cell recruitment by means of activation of afferents to the CeA. However, although many studies have investigated the origins of afferent inputs to the CeA, we do not know which of these also respond to IL-1beta. Therefore, to identify candidate neurons responsible for the recruitment of CeA cells by an immune challenge, we iontophoretically deposited a retrograde tracer, cholera toxin b-subunit (CTb), into the CeA of rats 7 days before systemic delivery of IL-1beta (1 mug/kg, i.a.). By using combined immunohistochemistry, we then quantified the number of Fos-positive CTb cells in six major regions known to innervate the CeA. These included the medial prefrontal cortex, paraventricular thalamus (PVT), ventral tegmental area, parabrachial nucleus (PB), nucleus tractus solitarius, and ventrolateral medulla. Our results show that after deposit of CTb into the CeA, the majority of double-labeled cells were located in the PB and the PVT, suggesting that CeA cell activation by systemic IL-1beta is likely to arise predominantly from cell bodies located in these regions. These findings may have significant implications in determining the central pathways involved in generating acute central responses to a systemic immune challenge. J. Comp. Neurol. 452:288-296, 2002. (C) 2002 Wiley-Liss, Inc.

Opposing roles for medial and central amygdala in the initiation of noradrenergic cell responses to a psychological stressor

Psychological stressors trigger the activation of medullary noradrenergic cells, an effect that has been shown to depend upon yet-to-be-identified structures located higher in the brain. To test whether the amygdala is important in this regard, we examined the effects of amygdala lesions on noradrenergic cell responses to restraint, and also looked at whether any amygdala cells that respond to restraint project directly to the medulla. Ibotenic acid lesions of the medial amygdala completely abolished restraint-induced Fos expression in A1 and A2 noradrenergic cells. In contrast, lesions of the central amygdala actually facilitated noradrenergic cell responses to restraint. Tracer deposits in the dorsomedial (but not ventrolateral) medulla retrogradely labelled many cells in the central nucleus of the amygdala, but none of these cells expressed Fos in response to restraint. These data suggest for the first time that the medial amygdala is critical to the activation of medullary noradrenergic cells by a psychological stressor whereas the central nucleus exerts an opposing, inhibitory influence upon noradrenergic cell recruitment. The initiation of noradrenergic cell responses by the medial amygdala does not involve a direct projection to the medulla. Accordingly, a relay through some other structure, such as the hypothalamic paraventricular nucleus, warrants careful consideration.

Determination of regional flow by use of intravascular PET tracers: Microvascular theory and experimental validation for pig livers

Today, the standard approach for the kinetic analysis of dynamic PET studies is compartment models, in which the tracer and its metabolites are confined to a few well-mixed compartments. We examine whether the standard model is suitable for modern PET data or whether theories including more physiologic realism can advance the interpretation of dynamic PET data. A more detailed microvascular theory is developed for intravascular tracers in single-capillary and multiple-capillary systems. The microvascular models, which account for concentration gradients in capillaries, are validated and compared with the standard model in a pig liver study. Methods: Eight pigs underwent a 5-min dynamic PET study after O-15-carbon monoxide inhalation. Throughout each experiment, hepatic arterial blood and portal venous blood were sampled, and flow was measured with transit-time flow meters. The hepatic dual-inlet concentration was calculated as the flow-weighted inlet concentration. Dynamic PET data were analyzed with a traditional single-compartment model and 2 microvascular models. Results: Microvascular models provided a better fit of the tissue activity of an intravascular tracer than did the compartment model. In particular, the early dynamic phase after a tracer bolus injection was much improved. The regional hepatic blood flow estimates provided by the microvascular models (1.3 +/- 0.3 mL min(-1) mL(-1) for the single-capillary model and 1.14 +/- 0.14 min(-1) mL(-1) for the multiple-capillary model) (mean +/- SEM mL of blood min(-1) mL of liver tissue(-1)) were in agreement with the total blood flow measured by flow meters and normalized to liver weight (1.03 +/- 0.12 mL min(-1) mL(-1)). Conclusion: Compared with the standard compartment model, the 2 microvascular models provide a superior description of tissue activity after an intravascular tracer bolus injection. The microvascular models include only parameters with a clear-cut physiologic interpretation and are applicable to capillary beds in any organ. In this study, the microvascular models were validated for the liver and provided quantitative regional flow estimates in agreement with flow measurements.

Impulse-response function of splanchnic circulation with model-independent constraints: theory and experimental validation

Modeling physiological processes using tracer kinetic methods requires knowledge of the time course of the tracer concentration in blood supplying the organ. For liver studies, however, inaccessibility of the portal vein makes direct measurement of the hepatic dual-input function impossible in humans. We want to develop a method to predict the portal venous time-activity curve from measurements of an arterial time-activity curve. An impulse-response function based on a continuous distribution of washout constants is developed and validated for the gut. Experiments with simultaneous blood sampling in aorta and portal vein were made in 13 anesthetized pigs following inhalation of intravascular [O-15] CO or injections of diffusible 3-O[ C-11] methylglucose (MG). The parameters of the impulse-response function have a physiological interpretation in terms of the distribution of washout constants and are mathematically equivalent to the mean transit time ( T) and standard deviation of transit times. The results include estimates of mean transit times from the aorta to the portal vein in pigs: (T) over bar = 0.35 +/- 0.05 min for CO and 1.7 +/- 0.1 min for MG. The prediction of the portal venous time-activity curve benefits from constraining the regression fits by parameters estimated independently. This is strong evidence for the physiological relevance of the impulse-response function, which includes asymptotically, and thereby justifies kinetically, a useful and simple power law. Similarity between our parameter estimates in pigs and parameter estimates in normal humans suggests that the proposed model can be adapted for use in humans.

Calcium treatment of harvested Geraldton waxflower does not enhance postharvest quality

Postharvest flower abscission from cut Geraldton waxflower (Chamelaucium uncinatum) is mostly caused by fungal invasion. Elevated plant tissue calcium concentrations through postharvest application reduces fungal disease severity in various crops. Such results may be explained by strengthening of plant cell walls by calcium. Strengthening provides a structural barrier to fungal hyphae, thereby restricting invasion of plant cells. Postharvest pulsing with calcium solution substantially increased calcium concentrations in waxflower tissues. Ca-45 tracer revealed calcium distribution throughout flowering sprigs, including infection sites such as stylar tissue. However, pulsing waxflower sprigs with calcium did not suppress either disease or flower abscission, nor did it enhance vase life.

The blood volumes of the primary and secondary circulatory system in the Atlantic cod Gadus morhua L., using plasma bound Evans Blue and compartmental analysis

The volume of the primary (PCS) and secondary (SCS) circulatory system in the Atlantic cod Gadus morhua was determined using a modified dye dilution technique. Cod (N=10) were chronically cannulated in the second afferent branchial artery with PE-50 tubing. Evans Blue dye was bound to harvested fish plasma at a concentration of 1 mg dye ml(-1) plasma, and injected at a concentration of 1 mg kg(-1) body mass. Serial sampling from the cannula produced a dye dilution curve, which could be described by a double exponential decay equation. Curve analysis enabled the calculation of the primary circulatory and total distribution volume. The difference between these volumes is assumed to be the volume of the SCS. From the dilution curve, it was also possible to calculate flow rates between and within the systems. The results of these experiments suggest a plasma volume in the PCS of 3.42+/-0.89 ml 100 g(-1) body mass, and in the SCS of 1.68+/-0.35 ml 100 g(-1) body mass (mean +/- S.D.) or approximately 50% that of the PCS. Flow rates to the SCS were calculated as 2.7% of the resting cardiac output. There was an allometric relationship between body mass and blood volumes. Increasing condition factor showed a tendency towards smaller blood volumes of the PCS, expressed as percentage body mass, but this was not evident for the volume of the SCS.