Transcript fusions as a result of chromosomal rearrangements have been a focus of attention in cancer as they provide attractive therapeutic targets. To identify novel fusion transcripts with the ...potential to be exploited therapeutically, we analyzed RNA sequencing, DNA copy number and gene mutation data from 4366 primary tumor samples. To avoid false positives, we implemented stringent quality criteria that included filtering of fusions detected in RNAseq data from 364 normal tissue samples. Our analysis identified 7887 high confidence fusion transcripts across 13 tumor types. Our fusion prediction was validated by evidence of a genomic rearrangement for 78 of 79 fusions in 48 glioma samples where whole-genome sequencing data were available. Cancers with higher levels of genomic instability showed a corresponding increase in fusion transcript frequency, whereas tumor samples harboring fusions contained statistically significantly fewer driver gene mutations, suggesting an important role for tumorigenesis. We identified at least one in-frame protein kinase fusion in 324 of 4366 samples (7.4%). Potentially druggable kinase fusions involving ALK, ROS, RET, NTRK and FGFR gene families were detected in bladder carcinoma (3.3%), glioblastoma (4.4%), head and neck cancer (1.0%), low-grade glioma (1.5%), lung adenocarcinoma (1.6%), lung squamous cell carcinoma (2.3%) and thyroid carcinoma (8.7%), suggesting a potential for application of kinase inhibitors across tumor types. In-frame fusion transcripts involving histone methyltransferase or histone demethylase genes were detected in 111 samples (2.5%) and may additionally be considered as therapeutic targets. In summary, we described the landscape of transcript fusions detected across a large number of tumor samples and revealed fusion events with clinical relevance that have not been previously recognized. Our results support the concept of basket clinical trials where patients are matched with experimental therapies based on their genomic profile rather than the tissue where the tumor originated.
Besides chemically interacting with hard tooth tissue, acidic functional monomers of self-etch adhesives should etch the prepared tooth surface to dissolve the smear layer and to provide surface ...micro-retention. Although the etching efficacy of functional monomers is commonly determined in terms of pH, the pH of adhesives cannot accurately be measured. Better is to measure the hydroxyapatite (HAp)–dissolving capacity, also considering that functional monomers may form monomer-Ca salts. Here, the etching efficacy of 6 functional monomers (GPDM, phenyl-P, MTEGP, 4-META, 6-MHP and 10-MDP) was investigated. Solutions containing 15 wt% monomer, 45 wt% ethanol, and 40 wt% water were prepared. Initially, we observed enamel surfaces exposed to monomer solution by scanning electron microscopy (SEM). X-ray diffraction (XRD) was employed to detect monomer-Ca salt formation. Phenyl-P exhibited a strong etching effect, while 10-MDP–treated enamel showed substance deposition, which was identified by XRD as 10-MDP–Ca salt. To confirm these SEM/XRD findings, we determined the etching efficacy of functional monomers by measuring both the concentration of Ca released from HAp using inductively coupled plasma–atomic emission spectroscopy (ICP-AES) and the amount of monomer-Ca salt formation using 31P magic-angle spinning (MAS) nuclear magnetic resonance (NMR). ICP-AES revealed that the highest Ca concentration was produced by phenyl-P and the lowest Ca concentration, almost equally, by 4-META and 10-MDP. Only 10-MDP formed 10-MDP–Ca salts, indicating that 10-MDP released more Ca from HAp than was measured by ICP-AES. Part of the released Ca was consumed to form 10-MDP–Ca salts. It is concluded that the repeatedly reported higher bonding effectiveness of 10-MDP–based adhesives must not only be attributed to the more intense chemical bonding of 10-MDP but also to its higher etching potential, a combination the other functional monomers investigated lack.
According to the ‘Adhesion–Decalcification’ concept, specific functional monomers within dental adhesives can ionically interact with hydroxyapatite (HAp). Such ionic bonding has been demonstrated ...for 10-methacryloyloxydecyl dihydrogen phosphate (MDP) to manifest in the form of self-assembled ‘nano-layering’. However, it remained to be explored if such nano-layering also occurs on tooth tissue when commercial MDP-containing adhesives (Clearfil SE Bond, Kuraray; Scotchbond Universal, 3M ESPE) were applied following common clinical application protocols. We therefore characterized adhesive-dentin interfaces chemically, using x-ray diffraction (XRD) and energy-dispersive x-ray spectroscopy (EDS), and ultrastructurally, using (scanning) transmission electron microscopy (TEM/STEM). Both adhesives revealed nano-layering at the adhesive interface, not only within the hybrid layer but also, particularly for Clearfil SE Bond (Kuraray), extending into the adhesive layer. Since such self-assembled nano-layering of two 10-MDP molecules, joined by stable MDP-Ca salt formation, must make the adhesive interface more resistant to biodegradation, it may well explain the documented favorable clinical longevity of bonds produced by 10-MDP-based adhesives.
Abstract 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.
X-ray diffraction (XRD) surface analysis and ultrastructural interfacial characterization using transmission electron microscopy (TEM) confirmed that the functional monomer 10–methacryloyloxydecyl ...dihydrogen phosphate (10-MDP) self-assembles into nano-layers at adhesive-tooth interfaces. Self-assembled nano-layering is thought to contribute to the durability of bonding to tooth dentin, although this has not been proven yet. In order to disclose this potential bond-durability contribution of nano-layering, we observed the 3-dimensional (3D) spreading of nano-layering by a series of focused-ion-beam (FIB) milled cross sections by scanning electron microscopy (FIB-SEM) and examined the mechanical properties of self-assembled nano-layering using scanning probe microscopy (SPM). A commercial 10-MDP-containing 3-step self-etch adhesive partially demineralized dentin up to submicron depth, forming a submicron hydroxyapatite-rich hybrid layer. TEM chemically and ultrastructurally confirmed the formation of interfacial nano-layering. FIB-SEM 3D reconstructions disclosed a 3D network of self-assembled nano-layering extending from the hybrid layer up to within the adjacent adhesive-resin layer. SPM revealed that nano-layering within the adhesive-resin layer possessed a higher elastic modulus than that of the surrounding adhesive resin, hereby suggesting that nano-layering contributes to the mechanical strength of adhesives like filler particles do. Nano-layering’s 3D expanded structure is expected to strengthen the surrounding resin, as well to better interconnect the adhesive-resin layer to the hybrid layer. In conclusion, this exploratory study demonstrated that nano-layering constitutes a strong phase at the adhesive interface, which may contribute to the clinical longevity of the 10-MDP-based bond to dentin.
Abstract This paper reflects on the state of the art of self-etch adhesives anno 2010. After presenting the general characteristics of self-etch adhesives, the major shortcomings of the most ...simple-to-use one-step (self-etch) adhesives are addressed. Special attention is devoted to the AD-concept and the benefit of chemical interfacial interaction with regard to bond durability. Finally, issues like the potential interference of surface smear and the more challenging bond to enamel for ‘mild’ self-etch adhesives are discussed.
As frequently added to adhesives, the mono-functional monomer 2-hydroxyethyl methacrylate (HEMA) acts as co-solvent and improves surface wetting. Nevertheless, HEMA promotes watersorption and ...hydrolysis at adhesive interfaces, affecting bond durability to dentin. This study investigated if two acrylamide co-monomer alternatives could replace HEMA in experimental adhesive-resin formulations as part of 3/2-step universal adhesives applied, respectively, in etch-and-rinse (E&R) and self-etch (SE) bonding modes.
Upon priming dentin with the 10-MDP-based Clearfil SE Bond 2’ primer (‘C-SE2p’; Kuraray Noritake), three experimental adhesive resins, consisting of 50 wt.% Bis-GMA, 15 wt.% TEGDMA, and either 35 wt.% diethyl acrylamide (‘DEAA’), hydroxyethyl acrylamide (‘HEAA’) or HEMA (‘HEMA+’), were applied. The control HEMA-free adhesive resin contained 60 wt.% Bis-GMA and 40 wt.% TEGDMA (‘HEMA−’). All adhesives were evaluated for ‘immediate’ and ‘aged’ micro-tensile bond strength (μTBS) to dentin upon, respectively, 1-week (1w) and 6-month (6m) water storage, TEM adhesive-dentin interfacial interaction, 24-h and 6m three-point bending, contact-angle wetting, viscosity and watersorption.
Linear mixed-effects model statistics revealed significantly better bonding performance of the adhesives applied in E&R than SE mode, except for DEAA_1w, with the highest μTBSs recorded for DEAA and HEMA− applied in SE mode. In E&R mode, aging did not significantly reduce DEAA’s μTBS. Best wetting on primed dentin was recorded for HEMA+, significantly better than DEAA, further HEAA and HEMA−, these directly related to their viscosity. HEAA absorbed significantly more water than all other adhesive-resin formulations. HEMA−>DEAA>HEAA>HEMA+ was the significant order for 6m bending strength.
The acrylamide co-monomer DEAA could replace HEMA, while HEAA not.
Ceramic restorations are often adhesively luted onto the tooth prep. The so-called touch-cure concept was developed to yield optimum polymerization of composite cement at the restoration-cement-tooth ...interface for immediate bond stabilization. Although this touch cure is theorized to initiate polymerization at the interface when the accelerator in the primer makes contact with the cement, this process has not yet been proven. This study aimed to elucidate the mechanism of touch cure by measuring the degree of conversion (DC) of composite cement applied with or without an accelerator-containing tooth primer (TP) versus an accelerator-free primer using real-time Fourier-transform infrared spectroscopy (RT-FTIR) and attenuated total reflection (ATR)–FTIR. Interfacial bond strength was measured in shear mode, the accelerator composition confirmed by X-ray fluorescence analysis (XRF), and the interfacial interaction of TP and composite cement with dentin investigated by X-ray diffraction (XRD), focused-ion-beam scanning electron microscopy (FIB-SEM) with 3-dimensional interface reconstruction, and transmission electron microscopy (TEM). RT/ATR-FTIR revealed the significantly highest DC when the composite cement was applied with the accelerator-containing primer. XRF disclosed a vanadium compound as a novel chemical accelerator within TP, instead of a classic chemical curing initiator system, to set off touch cure as soon the cement contacts the previously applied primer. Although the TP contains the acidic functional monomer 10-MDP for adhesion to tooth tissue, touch cure using the accelerator-containing TP combined the fastest/highest DC with the highest bond strength. FIB-SEM and TEM confirmed the tight interfacial interaction at dentin with submicron hybridization along with stable 10-MDP also Ca-salt nanolayering.
Previous research showed that the functional monomer 10-methacryloxydecyl dihydrogen phosphate (MDP) ionically bonds to hydroxyapatite (HAp) and forms a nano- layered structure at the interface with ...HAp-based substrates. Such hydrophobic nano-layering is considered to contribute to the long-term durability of the bond to tooth tissue. However, dental adhesives are complex mixtures usually containing different monomers. This study investigated the effect of the monomer 2-hydroxyethylmethacrylate (HEMA) on the chemical interaction of MDP with HAp by x-ray diffraction (XRD), nuclear magnetic resonance (NMR), and quartz crystal microbalance (QCM). We examined the chemical interaction of 5 experimental MDP solutions with increasing concentrations of HEMA. XRD revealed that addition of HEMA inhibits nano-layering at the interface, while NMR confirmed that MDP remained adsorbed onto the HAp surface. QCM confirmed this adsorption of MDP to HAp, as well as revealed that the demineralization rate of HAp by MDP was reduced by HEMA. It was concluded that even though the adsorption of MDP to HAp was not hindered, addition of HEMA inhibited interfacial nano-layering. Potential consequences with regard to bond durability necessitate further research.
Transcriptional profile-based subtypes of cancer are often viewed as identifying different diseases from the same tissue origin. Understanding the mechanisms driving the subtypes may be key in ...development of novel therapeutics but is challenged by lineage-specific expression signals. Using a t-test statistics approach, we compared gene expression subtypes across 12 tumor types, which identified eight transcriptional superclusters characterized by commonly activated disease pathways and similarities in gene expression. One of the largest superclusters was determined by the upregulation of a proliferation signature, significant enrichment in TP53 mutations, genomic loss of CDKN2A (p16(ARF)), evidence of increased numbers of DNA double strand breaks and high expression of cyclin B1 protein. These correlations suggested that abrogation of the P53-mediated apoptosis response to DNA damage results in activation of cell cycle pathways and represents a common theme in cancer. A second consistent pattern, observed in 9 of 11 solid tumor types, was a subtype related to an activated tumor-associated stroma. The similarity in transcriptional footprints across cancers suggested that tumor subtypes are commonly unified by a limited number of molecular themes.
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