Influence of resin composite insertion technique in preparations with a high C-factor

Objective: To evaluate the marginal microleakage in enamel and dentin/cementum walls in preparations with a high C-factor, using 3 resin composite insertion techniques. The null hypothesis was that there is no difference among the 3 resin composite insertion techniques. Method and Materials: Standardized Class 5 cavities were prepared in the lingual and buccal aspects of 30 caries-free, extracted third molars. The prepared teeth were randomly assigned to 3 groups: (1) oblique incremental placement technique, (2) horizontal incremental placement technique, and (3) bulk insertion (single increment). The preparations were restored with a 1-bottle adhesive (Single Bond, 3M ESPE) and microhybrid resin composite (Z100, 3M ESPE). Specimens were isolated with nail varnish except for a 2-mm-wide rim around the restoration and thermocycled (1,000 thermal cycles, 5°C/55°C; 30-second dwell time). The specimens were immersed in an aqueous solution of 50 wt% silver nitrate for 24 hours, followed by 8 hours in a photo-developing solution and evaluated for microleakage using an ordinal scale of 0 to 4. The microleakage scores obtained from occlusal and gingival walls were analyzed with Wilcoxon and Kruskal-Wallis nonparametric tests. Results: The null hypothesis was accepted. The horizontal incremental placement technique, the oblique incremental technique, and bulk insertion resulted in statistically similar enamel and dentin microleakage scores. Conclusion: Neither the incremental techniques nor the bulk placement technique were capable of eliminating the marginal microleakage in preparations with a high C-factor.

Nuclear mitochondrial-like sequences in ants: Evidence from Atta cephalotes (Formicidae: Attini)

Nuclear mitochondrial-like sequences (numts) are copies of mitochondrial DNA that have migrated to the genomic DNA. We present the first characterization of numts in ants, these numts being homologues to a mitochondrial DNA fragment containing loci the 3′ portion of the cytochrome oxidase I gene, an intergenic spacer, the tRNA leucine gene and the 5′ portion of the cytochrome oxidase II gene. All 67 specimens of Atta cephalotes (Hymenoptera: Formicidae: Attini) investigated had these homologues, which are within two monophyletic groups that we called numt1 and numt2. Numt1 and numt2 sequences are less variable than mitochondrial sequences and released from the severe purifying selection constraining the evolution of mitochondrial genes. Their formation probably involved bottlenecks related to two distinct transfer events of ancient and fast evolving mitochondrial DNA fragments to comparative slowly evolving nuclear DNA regions. © 2007 The Authors.

Report of a chimeric origin of transposable elements in a bovine-coding gene

Despite the wide distribution of transposable elements (TEs) in mammalian genomes, part of their evolutionary significance remains to be discovered. Today there is a substantial amount of evidence showing that TEs are involved in the generation of new exons in different species. In the present study, we searched 22,805 genes and reported the occurrence of TE-cassettes in coding sequences of 542 cow genes using the RepeatMasker program. Despite the significant number (542) of genes with TE insertions in exons only 14 (2.6%) of them were translated into protein, which we characterized as chimeric genes. From these chimeric genes, only the FAST kinase domains 3 (FASTKD3) gene, present on chromosome BTA 20, is a functional gene and showed evidence of the exaptation event. The genome sequence analysis showed that the last exon coding sequence of bovine FASTKD3 is ∼85% similar to the ART2A retrotransposon sequence. In addition, comparison among FASTKD3 proteins shows that the last exon is very divergent from those of Homo sapiens, Pan troglodytes and Canis familiares. We suggest that the gene structure of bovine FASTKD3 gene could have originated by several ectopic recombinations between TE copies. Additionally, the absence of TE sequences in all other species analyzed suggests that the TE insertion is clade-specific, mainly in the ruminant lineage. ©FUNPEC-RP.

Weak mixing and eigenvalues for arnoux-rauzy sequences

We define by simple conditions two wide subclasses of the socalled Arnoux-Rauzy systems; the elements of the first one share the property of (measure-theoretic) weak mixing, thus we generalize and improve a counterexample to the conjecture that these systems are codings of rotations; those of the second one have eigenvalues, which was known hitherto only for a very small set of examples.

Cementation of fiber post: influence of the cement insertion techniques on the bond strength of the fiber post-root dentin and the quality of the cement layer.

The aim of this study was to assess the influence of resin cement insertion methods on the bond strength of a fiber post to root dentin and quality of the cement layer. Forty bovine single-roots (length =16 mm) were randomly allocated into four groups, according to the cement insertion methods (N.=10): Gr1- Lentulo drill #40, Gr2- Centrix syringe, Gr3- Explorer #5, Gr4- fiber post. The root canals were prepared at 12 mm, using preparation bur # 3 of a cylinder quartz-FRC post (Aesthet post-plus, Bisco). The fiber posts were cemented using a multi-step etch-and-rinse adhesive system (All Bond 2®, Bisco) and a dual-cured resin cement (Duolink, Bisco). Each root was cut into seven samples: four samples of 1.8 mm thickness for push-out testing, and three with 0.5 mm for cement layer quality analyzing. One-way ANOVA was used for the push-out test values and the One-Way Kruskal-Wallis (P<0.05) and Dunn (10%) tests for the cement layer analyzes. ANOVA showed that the cement layer quality was affected by the cement insertion methods (P=0.0044): Gr1 (3.8 ± 1.3a), Gr2 (3.2 ± 1.3a), Gr3 (5.2 ± 1.5a,b) and Gr4 (5.2 ± 1.5b) (Dunn test), whereas the bond strength (MPa) was not affected by cement insertion methods: G1 (4.2 ± 1.3), G2 (3.2 ± 1.8), G3 (4.5 ± 0.9), G4 (3.1 ± 1.3). The fiber posts should be cemented with the assistance of the lentulo drill or centrix syringe to promote the best cement layer results.

Influence of high insertion torque on implant placement - an anisotropic bone stress analysis

The aim of this study was to evaluate the influence of the high values of insertion torques on the stress and strain distribution in cortical and cancellous bones. Based on tomography imaging, a representative mathematical model of a partial maxilla was built using Mimics 11.11 and Solid Works 2010 softwares. Six models were built and each of them received an implant with one of the following insertion torques: 30, 40, 50, 60, 70 or 80 Ncm on the external hexagon. The cortical and cancellous bones were considered anisotropic. The bone/implant interface was considered perfectly bonded. The numerical analysis was carried out using Ansys Workbench 10.0. The convergence of analysis (6%) drove the mesh refinement. Maximum principal stress (σ max) and maximum principal strain (ε max) were obtained for cortical and cancellous bones around to implant. Pearson's correlation test was used to determine the correlation between insertion torque and stress concentration in the periimplant bone tissue, considering the significance level at 5%. The increase in the insertion torque generated an increase in the σ max and ε max values for cortical and cancellous bone. The σmax was smaller for the cancellous bone, with greater stress variation among the insertion torques. The ε max was higher in the cancellous bone in comparison to the cortical bone. According to the methodology used and the limits of this study, it can be concluded that higher insertion torques increased tensile and compressive stress concentrations in the periimplant bone tissue.

Chromosome mapping of repetitive sequences in Anostomidae species: Implications for genomic and sex chromosome evolution

Background: Members of the Anostomidae family provide an interesting model system for the study of the influence of repetitive elements on genome composition, mainly because they possess numerous heterochromatic segments and a peculiar system of female heterogamety that is restricted to a few species of the Leporinus genus. The aim of this study was to isolate and identify important new repetitive DNA elements in Anostomidae through restriction enzyme digestion, followed by cloning, characterisation and chromosome mapping of this fragment. To identify repetitive elements in other Leporinus species and expand on studies of repetitive elements in Anostomidae, hybridisation experiments were also performed using previously described probes of LeSpeI repetitive elements. Results: The 628-base pair (bp) LeSpeII fragment was hybridised to metaphase cells of L. elongatus individuals as well as those of L. macrocephalus, L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii and S. isognathus. In L. elongatus, both male and female cells contained small clusters of LeSpeII repetitive elements dispersed on all of the chromosomes, with enrichment near most of the terminal portions of the chromosomes. In the female sex chromosomes of L. elongatus (Z2,Z2/W1W 2), however, this repeated element was absent. In the remaining species, a dispersed pattern of hybridisation was observed on all chromosomes irrespective of whether or not they were sex chromosomes. The repetitive element LeSpeI produced positive hybridisations signals only in L. elongatus, L. macrocephalus and L. obtusidens, i.e., species with differentiated sex chromosomes. In the remaining species, the LeSpeI element did not produce hybridisation signals. Conclusions: Results are discussed in terms of the effects of repetitive sequences on the differentiation of the Anostomidae genome, especially with respect to sex chromosome evolution. LeSpeII showed hybridisation patterns typical of Long Interspersed Elements (LINEs). The differential distribution of this element may be linked to sex chromosome differentiation in L. elongatus species. The relationship between sex chromosome specificity and the LeSpeI element is confirmed in the species L. elongatus, L. macrocephalus and L. obtusidens. © 2012 da Silva et al.; licensee BioMed Central Ltd.

Chromosomal evolution of neotropical cichlids: The role of repetitive DNA sequences in the organization and structure of karyotype

Cichlids are important in the aquaculture and ornamental fish trade and are considered models for evolutionary biology. However, most studies of cichlids have investigated African species, and the South American cichlids remain poorly characterized. Studies in neotropical regions have focused almost exclusively on classical cytogenetic approaches without investigating physical chromosomal mapping of specific sequences. The aim of the present study is to investigate the genomic organization of species belonging to different tribes of the subfamily Cichlinae (Cichla monoculus, Astronotus ocellatus, Geophagus proximus, Acaronia nassa, Bujurquina peregrinabunda, Hoplarchus psittacus, Hypselecara coryphaenoides, Hypselecara temporalis, Caquetaia spectabilis, Uaru amphiacanthoides, Pterophyllum leopoldi, Pterophyllum scalare, and Symphysodon discus) and reexamine the karyotypic evolutionary patterns proposed for this group. Variations in some cytogenetic markers were observed, although no trends were found in terms of the increase, decrease, or maintenance of the basal diploid chromosome number 2n = 48 in the tribes. Several species were observed to have 18S rDNA genetic duplications, as well as multiple rDNA loci. In most of the taxa analyzed, the 5S rDNA was located in the interstitial region of a pair of homologous chromosomes, although variations from this pattern were observed. Interstitial telomere sites were also observed and appear to be involved in chromosomal rearrangement events and the accumulation of repeat-rich satellite DNA sequences. Our data demonstrated the karyotypic diversity that exists among neotropical cichlids, suggesting that most of this diversity is due to the repetitive sequences present in heterochromatic regions and that repeat sequences have greatly influenced the karyotypic evolution of these fishes. © 2012 Springer Science+Business Media B.V.

Analysis of primary stability of dental implants inserted in different substrates using the pullout test and insertion torque

The aim of the study was to evaluate mechanical behavior of implants inserted in three substrates, by measuring the pullout strength and the relative stiffness. 32 implants (Master Porous-Conexao, cylindrical, external hexagon, and surface treatment) were divided into 4 groups (n = 8): pig rib bone, polyurethane Synbone, polyurethane Nacional 40 PCF, and pinus wood. Implants were installed with the exact distance of 5 mm of another implant. The insertion torque (N·cm) was quantified using the digital Kratos torque meter and the pullout test (N) was performed by an axial traction force toward the long axis of the implant (2 min/mm) through mount implant devices attached to a piece adapted to a load cell of 200 Kg of a universal testing machine (Emic DL10000). Data of insertion torque and maximum pullout force were submitted to one-way ANOVA and Bonferroni tests (α = 0.05). Polyurethane Nacional 40 PCF and pinus wood showed the highest values of insertion torque and pullout force, with significant statistical difference (P < 0.05) with other groups. The analysis showed stiffness materials with the highest values for primary stability. © 2013 Nathalia Ferraz Oliscovicz et al.

Composition and interrelationships of a large Neotropical freshwater fish group, the subfamily Cheirodontinae (Characiformes: Characidae): A case study based on mitochondrial and nuclear DNA sequences

•Relationships of Cheirodontinae based on a broad taxonomic sample.•Results reject the monophyly of Cheirodontinae as previously conceived.•Exclusion of Amazonspinther and Spintherobolus from the subfamily Cheirodontinae.•The removal of Leptagoniates pi of the genus Leptagoniates and inclusion in Cheirodontinae.•Division of Cheirodontinae in three newly defined monophyletic tribes. Characidae is the most species-rich family of freshwater fishes in the order Characiformes, with more than 1000 valid species that correspond to approximately 55% of the order. Few hypotheses about the composition and internal relationships within this family are available and most fail to reach an agreement. Among Characidae, Cheirodontinae is an emblematic group that includes 18 genera (1 fossil) and approximately 60 described species distributed throughout the Neotropical region. The taxonomic and systematic history of Cheirodontinae is complex, and only two hypotheses about the internal relationships in this subfamily have been reported to date. In the present study, we test the composition and relationships of fishes assigned to Cheirodontinae based on a broad taxonomic sample that also includes some characid incertae sedis taxa that were previously considered to be part of Cheirodontinae. We present phylogenetic analyses of a large molecular dataset of mitochondrial and nuclear DNA sequences. Our results reject the monophyly of Cheirodontinae as previously conceived, as well as the tribes Cheirodontini and Compsurini, and the genera Cheirodon, Compsura, Leptagoniates, Macropsobrycon, Odontostilbe, and Serrapinnus. On the basis of these results we propose: (1) the exclusion of Amazonspinther and Spintherobolus from the subfamily Cheirodontinae since they are the sister-group of all remaining Characidae; (2) the removal of Macropsobrycon xinguensis of the genus Macropsobrycon; (3) the removal of Leptagoniates pi of the genus Leptagoniates; (4) the inclusion of Leptagoniates pi in the subfamily Cheirodontinae; (5) the removal of Cheirodon stenodon of the genus Cheirodon and its inclusion in the subfamily Cheirodontinae under a new genus name; (6) the need to revise the polyphyletic genera Compsura, Odontostilbe, and Serrapinnus; and (7) the division of Cheirodontinae in three newly defined monophyletic tribes: Cheirodontini, Compsurini, and Pseudocheirodontini. Our results suggest that our knowledge about the largest Neotropical fish family, Characidae, still is incipient. © 2013 Elsevier Inc..

Tracking the evolution of sex chromosome systems in Melanoplinae grasshoppers through chromosomal mapping of repetitive DNA sequences

Background: The accumulation of repetitive DNA during sex chromosome differentiation is a common feature of many eukaryotes and becomes more evident after recombination has been restricted or abolished. The accumulated repetitive sequences include multigene families, microsatellites, satellite DNAs and mobile elements, all of which are important for the structural remodeling of heterochromatin. In grasshoppers, derived sex chromosome systems, such as neo-XY♂/XX♀ and neo-X1X2Y♂/X 1X1X2X2♀, are frequently observed in the Melanoplinae subfamily. However, no studies concerning the evolution of sex chromosomes in Melanoplinae have addressed the role of the repetitive DNA sequences. To further investigate the evolution of sex chromosomes in grasshoppers, we used classical cytogenetic and FISH analyses to examine the repetitive DNA sequences in six phylogenetically related Melanoplinae species with X0♂/XX♀, neo-XY♂/XX♀ and neo-X1X2Y♂/X1X1X 2X2♀ sex chromosome systems. Results: Our data indicate a non-spreading of heterochromatic blocks and pool of repetitive DNAs (C 0 t-1 DNA) in the sex chromosomes; however, the spreading of multigene families among the neo-sex chromosomes of Eurotettix and Dichromatos was remarkable, particularly for 5S rDNA. In autosomes, FISH mapping of multigene families revealed distinct patterns of chromosomal organization at the intra- and intergenomic levels. Conclusions: These results suggest a common origin and subsequent differential accumulation of repetitive DNAs in the sex chromosomes of Dichromatos and an independent origin of the sex chromosomes of the neo-XY and neo-X1X2Y systems. Our data indicate a possible role for repetitive DNAs in the diversification of sex chromosome systems in grasshoppers. © 2013Palacios-Gimenez et al.; licensee BioMed Central Ltd.

Orthogonal polynomials on the unit circle and chain sequences

Szego{double acute} has shown that real orthogonal polynomials on the unit circle can be mapped to orthogonal polynomials on the interval [-1,1] by the transformation 2x=z+z-1. In the 80's and 90's Delsarte and Genin showed that real orthogonal polynomials on the unit circle can be mapped to symmetric orthogonal polynomials on the interval [-1,1] using the transformation 2x=z1/2+z-1/2. We extend the results of Delsarte and Genin to all orthogonal polynomials on the unit circle. The transformation maps to functions on [-1,1] that can be seen as extensions of symmetric orthogonal polynomials on [-1,1] satisfying a three-term recurrence formula with real coefficients {cn} and {dn}, where {dn} is also a positive chain sequence. Via the results established, we obtain a characterization for a point w(|w|=1) to be a pure point of the measure involved. We also give a characterization for orthogonal polynomials on the unit circle in terms of the two sequences {cn} and {dn}. © 2013 Elsevier Inc.

Chromosomal organization of repetitive DNA sequences in Astyanax bockmanni (Teleostei, Characiformes): dispersive location, association and co-localization in the genome

Repetitive DNA sequences constitute a great portion of the genome of eukaryotes and are considered key components to comprehend evolutionary mechanisms and karyotypic differentiation. Aiming to contribute to the knowledge of chromosome structure and organization of some repetitive DNA classes in the fish genome, chromosomes of two allopatric populations of Astyanax bockmanni were analyzed using classic cytogenetics techniques and fluorescent in situ hybridization, with probes for ribosomal DNA sequences, histone DNA and transposable elements. These Astyanax populations showed the same diploid number (2n = 50), however with differences in chromosome morphology, distribution of constitutive heterochromatin, and location of 18S rDNA and retroelement Rex3 sites. In contrast, sites for 5S rDNA and H1, H3 and H4 histones showed to be co-located and highly conserved. Our results indicate that dispersion and variability of 18S rDNA and heterochromatin sites are not associated with macro rearrangements in the chromosome structure of these populations. Similarly, distinct evolutionary mechanisms would act upon histone genes and 5S rDNA, contributing to chromosomal association and co-location of these sequences. Data obtained indicate that distinct mechanisms drive the spreading of repetitive DNAs in the genome of A. bockmanni. Also, mobile elements may account for the polymorphism of the major rDNA sites and heterochromatin in this genus. © 2013 Springer Science+Business Media Dordrecht.

Triplet entropy analysis of hemagglutinin and neuraminidase sequences measures influenza virus phylodynamics

The influenza virus has been a challenge to science due to its ability to withstand new environmental conditions. Taking into account the development of virus sequence databases, computational approaches can be helpful to understand virus behavior over time. Furthermore, they can suggest new directions to deal with influenza. This work presents triplet entropy analysis as a potential phylodynamic tool to quantify nucleotide organization of viral sequences. The application of this measure to segments of hemagglutinin (HA) and neuraminidase (NA) of H1N1 and H3N2 virus subtypes has shown some variability effects along timeline, inferring about virus evolution. Sequences were divided by year and compared for virus subtype (H1N1 and H3N2). The nonparametric Mann-Whitney test was used for comparison between groups. Results show that differentiation in entropy precedes differentiation in GC content for both groups. Considering the HA fragment, both triplet entropy as well as GC concentration show intersection in 2009, year of the recent pandemic. Some conclusions about possible flu evolutionary lines were drawn. © 2013 Elsevier B.V.