Burden of itch in epidermolysis bullosa.

BACKGROUND: Itch is an unpleasant feeling that leads to scratching. It is a common, but understudied, problem in patients with epidermolysis bullosa (EB). OBJECTIVES: We measured the prevalence and characteristics of itch in the three major forms of EB: generalized EB simplex (EBS), junctional EB (JEB) and dystrophic EB (DEB). METHODS: Forty patients with EB were recruited from two tertiary care centres and one patient organization. The sample included 19 patients with EBS, seven with JEB and 14 with DEB. Patients completed the Leuven Itch Scale (LIS), a multidimensional self-report instrument that quantifies the frequency, duration, severity, distress, consequences and surface area of itch. This instrument has good clinimetric properties. RESULTS: Itch occurred in 85% of the patients, with substantial differences across the subtypes (EBS 74%, JEB 100%, DEB 93%). Itch, in all its dimensions, was most pronounced in patients with JEB and DEB, and less prominent in patients with EBS. The scores were significantly different for itch frequency, severity, distress and surface area, and the overall itch scores were comparable with those of atopic dermatitis. Itch mainly occurred in a hot environment (65%) and when sweating (62%). The most prevalent consequences were difficulty in falling asleep (88%) and lesions from scratching (85%). Differences between the three major subtypes were also observed in terms of circumstances, consequences and sensory characteristics. CONCLUSIONS: As expected, itch is common among patients with EB. All aspects of itch measured by the LIS were more severe in JEB and DEB than in EBS.

How much do resin-based dental materials release? A meta-analytical approach.

OBJECTIVES: Resin-based dental materials are not inert in the oral environment, and may release components, initially due to incomplete polymerization, and later due to degradation. Since there are concerns regarding potential toxicity, more precise knowledge of the actual quantity of released eluates is necessary. However, due to a great variety in analytical methodology employed in different studies and in the presentation of the results, it is still unclear to which quantities of components a patient may be exposed. The objective of this meta-analytical study was to review the literature on the short- and long-term release of components from resin-based dental materials, and to determine how much (order of magnitude) of those components may leach out in the oral cavity. METHODS: Out of an initial set of 71 studies, 22 were included. In spite of the large statistical incertitude due to the great variety in methodology and lack of complete information (detection limits were seldom mentioned), a meta-analytical mean for the evaluated eluates was calculated. To relate the amount of potentially released material components with the size of restorations, the mean size of standard composite restorations was estimated using a 3D graphical program. RESULTS: While the release of monomers was analyzed in many studies, that of additives, such as initiators, inhibitors and stabilizers, was seldom investigated. Significantly more components were found to be released in organic than in water-based media. Resin-based dental materials might account for the total burden of orally ingested bisphenol A, but they may release even higher amounts of monomers, such as HEMA, TEGDMA, BisGMA and UDMA. Compared to these monomers, similar or even higher amounts of additives may elute, even though composites generally only contain very small amounts of additives. A positive correlation was found between the total quantity of released eluates and the volume of extraction solution. SIGNIFICANCE: There is a clear need for more accurate and standardized analytical research to determine the long-term release from resin-based materials. Several guidelines for standardization are proposed.

Influence of the chemical structure of functional monomers on their adhesive performance.

Functional monomers in adhesive systems can improve bonding by enhancing wetting and demineralization, and by chemical bonding to calcium. This study tested the hypothesis that small changes in the chemical structure of functional monomers may improve their bonding effectiveness. Three experimental phosphonate monomers (HAEPA, EAEPA, and MAEPA), with slightly different chemical structures, and 10-MDP (control) were evaluated. Adhesive performance was determined in terms of microtensile bond strength of 4 cements that differed only for the functional monomer. Based on the Adhesion-Decalcification concept, the chemical bonding potential was assessed by atomic absorption spectrophotometry of the dissolution rate of the calcium salt of the functional monomers. High bond strength of the adhesive cement corresponded to low dissolution rate of the calcium salt of the respective functional monomer. The latter is according to the Adhesion-Decalcification concept, suggestive of a high chemical bonding capacity. We conclude that the adhesive performance of an adhesive material depends on the chemical structure of the functional monomer.

The role of HEMA in one-step self-etch adhesives.

UNLABELLED: In spite of its high allergenic potential, 2-hydroxyethyl methacrylate (HEMA), a low-molecular-weight monomer, is frequently used in adhesives for its positive influence on the bond strength. In addition, the presence of HEMA in one-component one-step adhesives can prevent phase separation. OBJECTIVES: In search of improved bonding effectiveness, the 24-h bond strength of four experimental one-step self-etch adhesives with different concentrations of HEMA to bur-cut enamel and dentin was determined using a micro-tensile bond strength protocol. METHODS: The tested experimental adhesives (Exp-0, Exp-10, Exp-19 and Exp-36) only differed in their concentration of HEMA, which was 0, 10, 19 and 36%, respectively. With an increasing concentration of HEMA, the concentration of acetone was decreased. Besides bond strength, the adhesives were also examined by light-microscopy for phase separation. The interface was investigated by SEM and TEM. RESULTS: Regarding bond strength, Exp-10 performed best. Even though Exp-36 was the only adhesive formulation that did not exhibit phase separation on a glass plate, it yielded the lowest bond strength. Accordingly, droplets could be observed by SEM and TEM in the adhesive layers of all adhesives, except for Exp-36 on enamel. CONCLUSION: A small amount of HEMA (10%) improved the bond strength of a one-step self-etch adhesive. When added in higher concentrations, this beneficial effect of HEMA on the bond strength is lost due to increased osmosis, which resulted in many droplets; due to reduced polymerization conversion; and sub-optimal physico-mechanical properties of the resultant poly-HEMA containing adhesive interface.

Origin of interfacial droplets with one-step adhesives.

Contemporary one-step self-etch adhesives are often documented with interfacial droplets. The objective of this study was to research the origin of these droplets. Two HEMA-rich and one HEMA-free adhesive were applied to enamel and dentin, with the lining composite either immediately cured or cured only after 20 min. All one-step adhesives exhibited droplets at the interface; however, the droplets had two different origins. With the HEMA-free adhesives, droplets were located throughout the adhesive layer and were stable in number over time. With the HEMA-rich adhesives, the droplets were observed exclusively at the adhesive resin/composite interface, and their number increased significantly when the composite was delay-cured. Only the latter droplets caused a significant drop in bond strength after delayed curing. While the droplets in the HEMA-free one-step adhesives should be ascribed to phase separation, those observed with HEMA-rich adhesives resulted from water absorption from dentin through osmosis.

Gel phase formation at resin-modified glass-ionomer/tooth interfaces.

Ionic bonding between polyalkenoic acid and hydroxyapatite may explain the excellent bonding retention of glass-ionomers in clinical trials. We have here investigated the extent to which the self-adhesiveness of resin-modified glass-ionomers (RMGIs) can be attributed to this chemical bonding capacity. Therefore, the interaction of 3 RMGIs with tooth substrates was comprehensively characterized, with electron and atomic force microscopy correlated with x-ray photoelectron spectroscopy (XPS). Interfacial ultrastructural analysis for 2 RMGIs disclosed a shallow hybridization of hydroxyapatite-coated collagen, on which a submicron gel phase was deposited through reaction of the polyalkenoic acid with calcium extracted from the dentin surface. One RMGI, however, bonded to dentin without hybrid layer or gel phase formation. XPS indicated that polycarboxylic acids included in the RMGIs electrostatically interacted with hydroxyapatite. We conclude that the self-adhesiveness of RMGIs should be attributed to ionic bonding to hydroxyapatite around collagen, and to micro-mechanical interlocking for those RMGIs that additionally hybridize dentin.

[The strategy of prevention and early detection of cutaneous melanoma]. / La stratégie de prévention et de dépistage du mélanome cutané.

Dermatology, and more specifically cutaneous oncology, has evolved from a descriptive science concept towards multifaceted medico-surgical medicine which is nowadays based not only on observation. In this view, the importance of prevention and early detection of skin cancers including cutaneous melanoma has been recognized and has created a new challenge. Thanks to this proactive approach, the improvement in the prognosis of newly diagnosed melanomas has been demonstrated, but many efforts have to be brought to reduce the incidence and the mortality of this potentially aggressive tumor.

Roughness and hydrophobicity studies of nanofiltration membranes using different modes of AFM.

Determination of the surface roughness by AFM is crucial to the study of particle fouling in nanofiltration. It is, however, very difficult to compare the different roughness values reported in the literature because of a lack in uniformity in the methods applied to determine surface roughness. AFM is used in both noncontact mode and tapping mode; moreover, the size of the scan area is highly variable. This study compares, for six different nanofiltration membranes (UTC-20, N30F, Desal 51HL, Desal 5DL, NTR7450, NF-PES-10), noncontact mode AFM with tapping mode AFM for several sizes of the scan area. Although the absolute roughness values are different for noncontact AFM and tapping mode AFM, no difference is found between the two modes of AFM in ranking the nanofiltration membranes with respect to their surface roughness. NTR 7450 and NF-PES-10 are the smoothest membranes, while the roughest surface can be found with Desal 51HL and Desal 5DL. UTC-20 and N30F are characterized by an intermediate roughness value. An increase in roughness with increasing scan area is observed for both AFM modes. Larger differences between the roughnesses of the membranes are obtained with tapping mode AFM because of the tapping of the tip on the surface. Phase imaging is an extension of tapping mode AFM, measuring the phase shift between the cantilever oscillation and the oscillation of the piezo driver. This phase shift reflects the interaction between the cantilever and the membrane surface. A comparison with contact angle measurements proves that a small phase shift corresponds to a large contact angle, representing a hydrophobic membrane surface.

Monomer-solvent phase separation in one-step self-etch adhesives.

One-step adhesives bond less effectively to enamel/dentin than do their multi-step versions. To investigate whether this might be due to phase separation between adhesive ingredients, we characterized the interaction of 5 experimental and 3 commercial self-etch adhesives with dentin using transmission electron microscopy. All adhesives were examined for homogeneity by light microscopy. Bonding effectiveness to dentin was determined with the use of a micro-tensile bond-strength protocol. The lower bond strength of the one-step adhesives was associated with light-microscopic observation of multiple droplets that disappeared slowly. Interfacial analysis confirmed the entrapment of droplets within the adhesive layer. The prompt disappearance of droplets upon application of a small amount of HEMA (2-hydroxyethyl methacrylate) or a HEMA-containing bonding agent, as well as the absence of droplets at the interface of all HEMA-containing adhesives, strongly suggests that the adhesive monomers separate from water upon evaporation of ethanol/acetone. Upon polymerization, the droplets become entrapped within the adhesive, potentially jeopardizing bond durability. This can be avoided by strong air-drying of the adhesive, thereby removing interfacial water and thus improving bonding effectiveness.

Adhesion to and decalcification of hydroxyapatite by carboxylic acids.

Fundamental to the processes of decalcification of or adhesion to mineralized tissues is the molecular interaction of acids with hydroxyapatite. This study was undertaken to chemically analyze the interaction of 1 mono-, 2 di-, 1 tri-, and 2 polycarboxylic acids with hydroxyapatite in an attempt to elucidate the underlying mechanism. Maleic, citric, and lactic acid decalcified hydroxyapatite, in contrast to oxalic acid and the two polycarboxylic acids that were chemically bonded to hydroxyapatite. Solubility tests showed that the calcium salts of the former were very soluble, whereas those of the latter could hardly be dissolved in the respective acid solutions. Based on these data, an adhesion/decalcification concept was advanced that predicts that carboxylic acids, regardless of concentration/pH, either adhere to or decalcify hydroxyapatite, depending on the dissolution rate of the respective calcium salts in the acid solution. This contrasting behavior of organic acids most likely results from their differential structural conformations.

Assessment of decontamination methods as pretreatment of silanization of composite glass fillers.

In terms of mechanical properties and durability, the surface of glass fillers should be decontaminated in order to optimize the silanization process for the production of resin composites. The objective of this study was to evaluate the decontamination efficiency of 18 cleaning methods on glass fillers as pretreatment of silane coupling. X-ray photoelectron spectroscopy revealed that SiO(2) boiled with a 5% sodium peroxodisulfate aqueous solution for 15 min, followed by ultrasonic rinsing with acetone for 30 min was most effective among all the decontamination methods investigated. In addition, nano-indentation measurements on SiO(2) treated by the above-mentioned method revealed that the surface was not significantly weakened as compared to untreated SiO(2). The results of this study should lead to an improved filler-matrix coupling and thus contribute to the development of better wear and fatigue-resistant composites. Therefore, sodium peroxodisulfate is proposed as a presilanization filler decontamination step in the production process of resin composites.

Evidence of chemical bonding at biomaterial-hard tissue interfaces.

For many years, glass-polyalkenoate cements have been described as possessing the unique properties of self-adherence to human hard tissues, such as bones or teeth. However, direct experimental evidence to prove the existence of chemical bonding has not been advanced. X-ray Photoelectron Spectroscopy (XPS) was used to analyze the chemical interaction of a synthesized polyalkenoic acid with enamel and synthetic hydroxyapatite. For both enamel and hydroxyapatite, the peak representing the carboxyl groups of the polyalkenoic acid was detected to have significantly shifted to a lower binding energy. De-convolution of this shifted peak disclosed two components with a peak representing unreacted carboxyl groups and a peak suggesting chemical bonding to hydroxyapatite. On average, 67.5% of the carboxyl groups of the polyalkenoic acid were measured to have bonded to hydroxyapatite. XPS of hydroxyapatite also disclosed its surface to be enriched in calcium and decreased in phosphorus, indicating that phosphorus was extracted at a relatively higher rate than calcium. Analysis of these data supports the mechanism in which carboxylic groups replace phosphate ions (PO4(3-)) of the substrate and make ionic bonds with calcium ions of hydroxyapatite. It is concluded that an ultrathin layer of a polyalkenoic acid can be prepared on a hydroxyapatite-based substrate by careful removal of non-bonded molecules. With this specimen-processing method, XPS not only provided direct evidence of chemical bonding, but also enabled us to quantify the percentages of functional groups of the polyalkenoic acids that bonded to calcium of hydroxyapatite.

A novel approach to AFM characterization of adhesive tooth-biomaterial interfaces.

A novel approach is proposed for studying tooth-biomaterial interactions with high resolution. Thus far, polished interfaces examined by AFM have not disclosed much detail, mainly due to the destruction of soft surface texture and the smearing of polishing debris across the interface that obscures the actual ultra-structure. Therefore the practical utility of diamond-knife microtomy as a sample preparation technique for imaging tooth-biomaterial interfaces by AFM with high resolution was tested in this study and compared to that of ultra-fine mechanical polishing techniques. The AFM images clearly demonstrated the enhanced potential of diamond-knife microtomy for nondestructively producing clean cross-sections through interfaces that allow the interfacial ultra-structure to be imaged by AFM with a resolution equaling that of TEM. This novel approach opens the field to the full range of scanning probe microscopy, including physical and chemical surface characterization of interfaces with a mix of soft and hard substrates.

Hybridization effectiveness of a two-step versus a three-step smear layer removing adhesive system examined correlatively by TEM and AFM.

PURPOSE: The objectives of this study were (1) to compare the hybridization effectiveness of two adhesive systems that are applied in respectively three and two steps, and (2) to determine the best resin-dentin interface preparation technique for atomic force microscopy (AFM). MATERIALS AND METHODS: The resin-dentin interface produced by the three-step OptiBond Dual-Cure (Kerr) and its simplified two-step successor OptiBond Solo (Kerr) was ultramorphologically examined using transmission electron microscopy (TEM) and AFM. Four different methods were used to prepare interface specimens for AFM: (1) polishing to a 0.1-micron finish with a silicon oxide suspension, (2) polishing to a 0.05-micron finish with an aluminum oxide suspension, (3) argon-ion etching, and (4) sectioning with a diamond knife. RESULTS: Both TEM and AFM demonstrated that some collapse of the exposed collagen fibril network, due to gentle postconditioning air-drying of the dentin surface, may not have been totally recovered through hybridization by the two-step adhesive formulation as opposed to the three-step precursor. From the four interface preparation methods, only diamond-knife sectioning revealed sufficient ultramorphologic detail and high resolution that can capitalize on the high resolution offered by AFM. CONCLUSION: First, the findings suggest that simplifying the application procedure of adhesives by combining the primer and adhesive resin into a single application step may reduce hybridization effectiveness. Future research should confirm this effect for other two- versus three-step adhesive systems. Second, diamond-knife sectioning should be used for future topographic imaging and physicomechanical testing of resin-dentin interfaces by AFM.

Strong enhancement of nonlinear optical properties through supramolecular chirality

A new approach to second-order nonlinear optical (NLO) materials is reported, in which chirality and supramolecular organization play key roles. Langmuir-Blodgett films of a chiral helicene are composed of supramolecular arrays of the molecules. The chiral supramolecular organization makes the second-order NLO susceptibility about 30 times larger for the nonracemic material than for the racemic material with the same chemical structure. The susceptibility of the nonracemic films is a respectable 50 picometers per volt, even though the helicene structure lacks features commonly associated with high nonlinearity. Susceptibility components that are allowed only by chirality dominate the second-order NLO response.

Imaging red blood cells with the atomic force microscope.

A novel technique for the reproduction of ultramorphological images and details of the surface of normal and pathological red blood cells (RBC) was investigated. The atomic force microscope (AFM) provided high-resolution images of cell surfaces. The RBC dimensions obtained by this technique revealed differences between native red cells in smears and glutaraldehyde-fixed red cells. It was shown that fixed red blood cells were best suited for the ultramorphological imaging of the cell surface.

Imaging uremic red blood cells with the atomic force microscope.

Ultramorphological images and details of an uremic echinocyte are shown. The images confirm the existence of uremic echinocytes and demonstrate that atomic force microscopy can provide high-resolution images of cell surfaces.

Protein engineering of xylose (glucose) isomerase from Actinoplanes missouriensis. 2. Site-directed mutagenesis of the xylose binding site.

Site-directed mutagenesis in the active site of xylose isomerase derived from Actinoplanes missouriensis is used to investigate the structural and functional role of specific residues. The mutagenesis work together with the crystallographic studies presented in detail in two accompanying papers adds significantly to the understanding of the catalytic mechanism of this enzyme. Changes caused by introduced mutations emphasize the correlation between substrate specificity and cation preference. Mutations in both His 220 and His 54 mainly affect the catalytic rate constant, with catalysis being severely reduced but not abolished, suggesting that both histidines are important, but not essential, for catalysis. Our results thus challenge the hypothesis that His 54 acts as an obligatory catalytic base for ring opening; this residue appears instead to be implicated in governing the anomeric specificity. With none of the active site histidines acting as a catalytic base, the role of the cations in catalyzing proton transfer is confirmed. In addition, Lys 183 appears to play a crucial part in the isomerization step, by assisting the proton shuttle. Other residues also are important but to a lesser extent. The conserved Lys 294 is indirectly involved in binding the activating cations. Among the active site aromatic residues, the tryptophans (16 and 137) play a role in maintaining the general architecture of the substrate binding site while the role of Phe 26 seems to be purely structural.