Temporal and spatial transcriptional programs in murine kidney development.

We have performed a systematic temporal and spatial expression profiling of the developing mouse kidney using Compugen long-oligonucleotide microarrays. The activity of 18,000 genes was monitored at 24-h intervals from 10.5-day-postcoitum (dpc) metanephric mesenchyme (MM) through to neonatal kidney, and a cohort of 3,600 dynamically expressed genes was identified. Early metanephric development was further surveyed by directly comparing RNA from 10.5 vs. 11.5 vs. 13.5dpc kidneys. These data showed high concordance with the previously published dynamic profile of rat kidney development (Stuart RO, Bush KT, and Nigam SK. Proc Natl Acad Sci USA 98: 5649-5654, 2001) and our own temporal data. Cluster analyses were used to identify gene ontological terms, functional annotations, and pathways associated with temporal expression profiles. Genetic network analysis was also used to identify biological networks that have maximal transcriptional activity during early metanephric development, highlighting the involvement of proliferation and differentiation. Differential gene expression was validated using whole mount and section in situ hybridization of staged embryonic kidneys. Two spatial profiling experiments were also undertaken. MM (10.5dpc) was compared with adjacent intermediate mesenchyme to further define metanephric commitment. To define the genes involved in branching and in the induction of nephrogenesis, expression profiling was performed on ureteric bud (GFP+) FACS sorted from HoxB7-GFP transgenic mice at 15.5dpc vs. the GFP- mesenchymal derivatives. Comparisons between temporal and spatial data enhanced the ability to predict function for genes and networks. This study provides the most comprehensive temporal and spatial survey of kidney development to date, and the compilation of these transcriptional surveys provides important insights into metanephric development that can now be functionally tested.

Seventeen-year clinical study of ultraviolet-cured posterior composite Class I and II restorations.

OBJECTIVES: To compare the clinical performance of four commercial ultraviolet light-cured composite materials, and to evaluate curing-system effects on long-term wear resistance of Class I and II restorations. MATERIALS AND METHODS: Approximately 32 samples of each of four different ultraviolet light-cured composites (n = 130) were inserted into conventional Class I and II cavity preparations by two clinicians. Cavosurface margins of the preparations were not beveled. Enamel walls of the preparation were etched, and the respective bonding agent was applied. Each restoration was evaluated by two clinicians at 5, 10, and 17 years. Direct evaluations were performed using modified United States Public Health Service (USPHS) criteria. Indirect evaluations were performed using the Leinfelder cast evaluation method. RESULTS: After 17 years, 65% of the restorations were recalled and pooled direct evaluations were conducted for color matching (94% alfa), marginal discoloration (100% alfa), marginal integrity (100% alfa), secondary caries (92% alfa), surface texture (72% alfa), and anatomic form (22% alfa). Mean occlusal wear from indirect evaluations at 5, 10, and 17 years was 197 +/- 85 microns, 235 +/- 72 microns, and 264 +/- 80 microns, respectively. For direct and indirect evaluations there were significant differences (p < or = .05) between the baseline and 5-year recall evaluations. CLINICAL SIGNIFICANCE: This study demonstrated that the mean pooled occlusal wear of four ultraviolet light-cured posterior composites at 17 years was 264 microns (approximately 0.25 mm), and that most wear (75%) occurred in the first 5 years. Of all recalled restorations, 76% were judged clinically acceptable at 17 years, and 22% of those exhibited no clinically detectable wear.

Relationship between filler and matrix resin characteristics and the properties of uncured composite pastes.

The relationships between filler type, filler content, matrix resin composition and viscosity and the flow characteristics of composite paste formulations have been investigated. BIS-GMA and 10 experimental BIS-GMA analogues were diluted as needed with TEGDMA to produce 1000 and 2000 cp solutions corresponding to the viscosity range of commercial composite matrices. All of these resins were mixed with a silanated hybrid-filler and a silanated micro-filler. For each combination the maximum filler content was determined, as well as the paste consistencies (plasticities) at a series of filler contents. Paste consistencies were measured by a modification of the ADA specification methods developed for determining standard test consistencies for zinc phosphate and silicate cements. For each monomer-filler combination increasing filler contents resulted in monotonically reduced plasticities. The maximum filler contents appeared to be an attribute of the particular filler rather than matrix resin characteristics. For the materials used, the maximum contents were approximately 86 wt% (83 vol%) filler for the hybrid filler, and 36% wt% (24 vol%) for the microfilled material. At lower filler contents, plasticity differences unexpectedly existed even at equal filler contents and matrix viscosities. Evidence was found that the plasticity varied both with the base monomer composition and the amount of TEGDMA present. The results imply that compositional interactions between the filler and matrix influence the results when all known mechanical factors are controlled.

Characterization of some aromatic dimethacrylates for dental composite applications.

Several novel dimethacrylates have been developed as alternative matrix materials for dental composite applications. For the cured bulk polymers the equilibrium water uptake, reduction of glass transition temperatures (Tgs) by water sorption, refractive indices and the surface hardness have been determined. The properties were then compared with those of the control Bis-GMA. These properties correlated well with the structures of the polymers. Polar groups were found to increase the water sorption and thus reduce surface hardness.

Structure property relationship among novel dental composite matrix resins.

Several novel dimethacrylate monomers of propoxylated diphenols have been synthesized to establish a correlation between their structure and the important properties of viscosity, curing shrinkage and wetting behaviour. These dimethacrylates were designed to possess linear and flexed structures. These new monomers exhibited very much lower viscosities compared to the conventionally used Bis-GMA. Good correlations were obtained for variations of the monomer structures with bulk viscosities and curing shrinkages. However, the wetting behaviour of these resins did not show any trend.

Water uptake of soft lining materials from osmotic solutions.

OBJECTIVES: The water uptake characteristics of soft lining materials are of obvious importance in that they are expected to function in the oral environment. Results for Novus (Hygenic Corp., Akron, OH, USA) show a very high uptake from distilled water. Despite this high uptake, Novus appears to function satisfactorily in the mouth. High water uptake of soft lining materials has been attributed to the presence of water soluble impurities that, on immersion, form solution droplets; the driving force for the uptake being the osmotic gradient between the droplets and the external solution. Uptake should therefore be less from ionic solutions. The object of this study was to test the applicability of this theory to Novus and two experimental soft lining materials. METHODS: Water uptake of two experimental materials and Novus has been determined from distilled water and two saline solutions (0.45 and 0.9 M). After 196 days specimens were desorbed to constant weight and then subjected to a second sorption cycle. RESULTS: Novus had the highest uptake from distilled water at approximately 18%, the experimental materials having an uptake approximately 7%. Desorptions were all rapid, minimum weight being reached within 1-2 days. Uptakes of the second sorptions from water were all higher. Uptake from saline solutions was approximately 12% for all materials, uptake from 0.9 M saline being the lowest. Second sorption results from solution were similar to the first. CONCLUSION: The results obtained support the theory that the high water uptake of elastomeric materials is osmotically driven.

Influence of hydrogen bonding on properties of BIS-GMA analogues.

The influence of chemical structure on the important properties of composite matrix resins is being systematically investigated. This study addresses the relationships between pendent side chain structures, viscosity and curing shrinkage. In particular, viscosity is known to be greatly influenced by intermolecular interactions, such as hydrogen bonding, and free volume effects. In order to establish the relative importance of these factors, analogues of BIS-GMA were synthesized in which the pendent hydroxyl groups were replaced by trimethyl siloxyl, and by dimethyl, isopropyl siloxyl groups. The viscosities were determined with a cone and plate viscometer and curing shrinkages were determined gravimetrically. They were compared to previously determined values for BIS-GMA and its methyl and hydrogen substituted analogues. The high viscosity of BIS-GMA is drastically reduced by replacement of the hydroxyl group, or its substitution by silylation. The relatively smaller effects produced by varying the bulk of the substituted side chains indicates that the main effect on viscosity is due to the presence or absence of hydrogen bonding. Conversely, increasing the bulk of the side chain substituent has less effect on viscosity, but significantly reduces the curing shrinkage. Changes in curing shrinkages were explained in terms of effects of free volumes associated with the monomers.

Effect of dilution on the kinetics of cross-linking thermal polymerization of dental composite matrix resins.

The effect of dilution of 2,2-bis-(4-(2-hydroxy-3-methacryloxyprop-1oxy) phenyl) propane (bis-GMA) with triethylene glycol demethacrylate (TEGDMA) on the extent of polymerization, Ep, has been studied using differential scanning calorimetry. The isothermal and dynamic measurements indicate that the Ep is affected dramatically by dilution. The residual polymerization was also found to be strongly influenced by dilution. However, the apparent activation energy, Eapp, was found to be independent of dilution.

Phosphorylation of the C-terminal domain of RNA polymerase II by the extracellular-signal-regulated protein kinase ERK2.

Rat ERK2, an extracellular-signal-regulated protein kinase family member, phosphorylates RNA polymerase II in vitro. Phosphorylation occurs within the heptapeptide repeats of the C-terminal domain of the largest subunit, in a region important for regulation of transcriptional activity. Analysis of deletion mutants and synthetic peptides showed that ERK2 phosphorylation occurs at multiple serine residues throughout the C-terminal domain, with no marked preference for consensus repeats versus naturally occurring variants. Our results are consistent with the idea that protein kinases in the extracellular-signal-regulated protein kinase family regulate transcription by direct phosphorylation of RNA polymerase II, but do not support a model where particular portions of the C-terminal domain are special targets of ERK phosphorylation.

Pooling of long term clinical wear data for posterior composites.

Clinical studies to evaluate the wear of posterior composite restorations are complicated by the large number of factors which influence the findings. A multi-factorial equation has been developed which successfully normalizes the effects of these factors within studies. This equation is not capable of normalizing these effects for other investigations reported in the literature because study characteristics essential to the analysis are rarely reported. Quantitative estimates of wear rate often differ dramatically between studies and between groups of investigators. The objective of this study was to investigate a potential solution to this problem by ranking materials within studies, using common materials to relate rankings across studies, and achieving an overall pooled ranking for products. An intensive literature search disclosed 78 articles and 46 abstracts on clinical posterior composite wear. All studies were analyzed which involved: (1) more than five restorations per material, (2) Class I or II restorations in posterior adult teeth, (3) characterized commercial products, (4) 2 or 3-year wear data, and (5) information on more than one composite material per study. There were 10 2-year studies involving 25 materials, and 10 3-year studies with 26 materials. Within each study the materials were ranked by wear, and the rankings were converted to centered modified ridits. A meta-analysis combining the data across studies was conducted using ANOVA. Although caution is needed for interpreting significance levels because of the small numbers of products evaluated per study, a high level of agreement in rank correlation of 28 products across qualifying studies was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Confirmation of Leinfelder clinical wear standards.

OBJECTIVES: Accuracy of composite wear studies based on Leinfelder standards has been disputed. There are differences with other well-calibrated systems such as the M-L and Vivadent wear standards. The objective of this study was to reevaluate the margin height at key regions along the restoration margins for each of the 6 Leinfelder standards using laser profiling techniques. METHODS: The Leinfelder standards were profiled in parallel paths 100 microns apart and measured in x-y-z position every 20 microns along those paths using a laser profilometer. RESULTS: Rounding of cavosurface enamel margins from intraoral wear greatly increased the uncertainty of the true enamel margin location and step height measurements, precluding unequivocal measurements for standards #2 and #3. Values for other standards for the original report, newly measured means and standard errors, and measured ranges were: #4 (322 microns, 333 +/- 34 microns, 171-507 microns), #5 (382 microns, 459 +/- 44 microns, 202-649 microns), and #6 (493 microns, 584 +/- 91 microns, 315-1022 microns). There were no statistically significant differences (p < or = 0.10) between these and original values. Large standard errors may have obscured small differences that may exist. The Leinfelder cast conversion scale seems to be the correct relative magnitude. SIGNIFICANCE: Differences between Leinfelder casts and other standards may be due to differences in shadow production. Clinical wear may be systematically underestimated by other cast evaluation methods that have well-defined margins. This emphasizes the need for standard casts with margin morphology similar to the clinical casts being evaluated for wear.

Protection hypothesis for composite wear.

Significant occlusal wear occurs on posterior composite restorations in areas without direct contact with opponent teeth (contact-free wear) in addition to wear at the contacts. Recent research has indicated that all occlusal posterior composite surfaces wear, and those surfaces show decreasing wear rates over time. This provides more evidence for an earlier theory proposed by Jørgensen et al. (1979) that composite contact-free wear is a function of access and attrition caused by small particles in the food bolus. This hypothesis is now called the "protection theory" or "protection hypothesis" for wear. Macroscopic protection or "sheltering" from wear is provided by cavity preparation walls. Microscopic protection against resin matrix wear is provided by filler particles that are close together. Clinical evidence supporting the protection hypothesis comes from recently confirmed low wear rates for microfill and hybrid composites. The objective of this work was to calculate the minimum inter-particle spacing required for microscopic protection (wear resistance) for composites based on assumptions of packing fractions (0.68) with small particles (0.02 microns radius) for dispersed versus agglomerated conditions. An inter-particle spacing (IPS) of < 0.10 microns was assumed to afford wear protection. The results are that only 1.5-6.0 volume percentage microfiller is theoretically required to generate an IPS of 0.1 micron for composite microscopic protection. However, microfiller particles are suspected to agglomerate into clusters. Modified calculations for that effect indicate that at least 35% filler is required to provide microscopic protection. This effect explains much of the 3 to 5 year clinical research results for microfills and hybrids.(ABSTRACT TRUNCATED AT 250 WORDS)

Five-year clinical study of u.v.-polymerized posterior composites.

Four u.v.-polymerized composites (Estilux, Nuva-Fil, Nuva-Fil P.A. and Uvio-Fil) were clinically evaluated in Class I and II cavity preparations in permanent teeth. The restorations were evaluated at baseline, 1, 2, 3, 4 and 5 years using USPHS criteria for direct evaluation and at 3, 4 and 5 years using the Leinfelder method for indirect evaluation of wear. The wear measured by the USPHS method showed increasing wear rates in earlier reports, apparently due to the limited sensitivity of the scale to early wear. The wear by the Leinfelder method showed gradually decreasing wear rates. There were no statistically significant differences among the wear values for the four u.v.-cured materials or between these materials and the self-cured and visible light-cured controls.

Correlation of M-L, Leinfelder, and USPHS clinical evaluation techniques for wear.

Most recent clinical research studies on wear of posterior composite restorations have used the USPHS and Leinfelder measuring techniques. New studies are adopting the Moffa-Lugassy (M-L) technique. However, to date there has been no means of comparing results of the different techniques. The objective of this study was to correlate the M-L scale wear values to Leinfelder scale wear values and to USPHS scale alfa-bravo transitions, so that data can be pooled across studies for comparisons. M-L evaluations were made for 221 restorations previously evaluated by the USPHS and Leinfelder methods. The M-L wear values were determined as the mean of three independent evaluators' values. At the USPHS alfa-bravo transition, the mean Leinfelder wear value was 192 microns, and the mean M-L value was 97 microns. The correlation coefficient for the linear regression of M-L values vs. Leinfelder values was 0.87. The scales were linearly related, with a slope of 0.52. The average values for M-L readings of wear were statistically different and approximately half the Leinfelder values.

Biomechanics of Ender rods, compression screw, and Zickel nail in the fixation of stable subtrochanteric femur osteotomies.

Compression screws, Ender rods, and Zickel nails were evaluated mechanically to determine their resulting stability in the fixation of experimental subtrochanteric femur osteotomies. Twenty-five pairs of human cadaver femurs were used. Experimental osteotomies were created and a random member of each pair was fixed with a compression screw (AMBI system, Richards Mfg. Co). The other member was fixed with either Ender rods or a Zickel nail. The specimens were oriented in a direction to approximate the in vivo position of maximum resultant force on the femur and then subjected to a single progressive load to failure on a materials testing machine. The ultimate strength of fixation with the compression screw was approximately 4.6 times greater than with the Ender rods and 1.8 times greater than that with the Zickel nail. Ender rods and Zickel nails failed primarily in external rotation, whereas the compression screw failed by bone fracturing and varus bending.

Reactivity of cysteine 18 in oncomodulin.

The reactivity of recombinant and tumor-derived preparations of oncomodulin toward 5,5'-dithiobis-(2-nitrobenzoate) (Ellman's reagent) and dansylaziridine was investigated. In contrast to previously published data (Mutus, B., Palmer, E. J., and MacManus, J. P. (1988) Biochemistry 27, 5615-5622), the apoprotein was observed to react far more rapidly than the calcium-bound form with Ellman's reagent. Attempts to quantitatively label the native protein with dansylaziridine met with little success, either with the metal-free or calcium-bound forms. In neither case did the extent of modification approach the level observed with the sodium dodecyl sulfate-denatured form of the protein. These results suggest that access to the sulfhydryl group of Cys-18 is severely restricted in the native protein, particularly when the high affinity ion-binding sites are occupied. Consistent with these observations, prolonged incubation of native oncomodulin at room temperature in the absence of reductant did not result in the generation of disulfide-linked dimers, either in the presence or absence of Ca2+. Interestingly, however, Cu2+ ion was observed to facilitate the apparent dimerization of oncomodulin. This reaction, which occurs more rapidly with the Ca2(+)-free form of the protein, affords material with the expected electrophoretic mobility. However, in contrast with the results of Mutus et al., dimeric oncomodulin prepared in this manner fails to stimulate bovine heart cAMP phosphodiesterase.

Comparison of direct and indirect methods for analyzing wear of posterior composite restorations.

The vast majority of recent clinical research involving wear analyses of posterior composite resin restorations have used either the direct evaluation method (USPHS) or the indirect cast comparison (Leinfelder) method. However, there has never been any established correlation of the two wear scales. The objective of this study was to determine the amount of wear for the USPHS alfa-bravo transition on the basis of cast comparison data. Clinical wear data were collected over five years by both the direct method and the indirect method. Three materials were used involving a total of 221 restorations in 61 patients. Each restoration was evaluated at baseline, six months, one, two, three, and five years by each method. Then, for those restorations which underwent a transition from an alfa to a bravo clinical rating, the absolute wear at the transition was determined by averaging the indirect ratings just before and just after the transition. The mean wear corresponding to the alfa-bravo transition was 192 +/- 60 microns. Because of the large width of the alfa category up to the alfa-bravo transition, extensive early changes and high initial wear rates cannot be detected by the USPHS system.

Three-year clinical evaluation of a self-cured posterior composite resin.

A study was conducted to determine whether the wear resistance of a posterior composite could be improved by maximizing filler particle-to-particle contacts. This was expected to reduce stress concentrations on the resin matrix and thus reduce occlusal wear. A self-curing quartz-filled composite with this design, P-10, was used to restore 90 Class I and II cavity preparations in adult teeth. Restorations were recalled after baseline at six months, one year, two years, and three years to measure wear by direct and indirect evaluation methods. There was no apparent advantage for this material compared with other previously evaluated posterior composites. The average cumulative wear for P-10 after three years was 145 microns. In addition, the restorations were evaluated for color-matching, interfacial staining, secondary caries, marginal adaptation, surface texture, and postoperative sensitivity. This material was not significantly different in those ways from other posterior composite products except in terms of more rapid color change, because it is self-cured.