Three-dimensional analysis for detection of pulp stones in a Saudi population using cone beam computed tomography.

PURPOSE: To assess the prevalence and distribution of pulp stones in a Saudi population. METHODS: A cone beam computed tomography (CBCT) analysis of 150 upper and 150 lower dental arches was performed. The relationships between pulp stones and age, sex, tooth type, dental arch, orthodontic treatment, caries, and restoration were statistically examined (P < 0.05). RESULTS: A total of 295 dental arches (98.3%) had at least one pulp stone, and the prevalence was higher in the maxillary teeth (56.2%) than in the mandibular teeth (48.1%) (P = 0.0003). Men were more likely to have pulp stones than women (P = 0.011 for the maxilla, P < 0.0001 for the mandible). Furthermore, age and orthodontic treatment had no significant effects on the incidence of pulp stones (P > 0.05). A higher occurrence of pulp stones was observed in the first molars (>91%), and in carious and restored teeth when compared to intact teeth (P < 0.05). CONCLUSION: Pulp stones were more prevalent in the upper dental arches, first molars, and carious and restored teeth, as well as in the male population, and were not associated with age or orthodontic treatment.

Phage Diving: An Exploration of the Carcharhinid Shark Epidermal Virome.

The epidermal microbiome is a critical element of marine organismal immunity, but the epidermal virome of marine organisms remains largely unexplored. The epidermis of sharks represents a unique viromic ecosystem. Sharks secrete a thin layer of mucus which harbors a diverse microbiome, while their hydrodynamic dermal denticles simultaneously repel environmental microbes. Here, we sampled the virome from the epidermis of three shark species in the family Carcharhinidae: the genetically and morphologically similar Carcharhinus obscurus (n = 6) and Carcharhinus galapagensis (n = 10) and the outgroup Galeocerdo cuvier (n = 15). Virome taxonomy was characterized using shotgun metagenomics and compared with a suite of multivariate analyses. All three sharks retain species-specific but highly similar epidermal viromes dominated by uncharacterized bacteriophages which vary slightly in proportional abundance within and among shark species. Intraspecific variation was lower among C. galapagensis than among C. obscurus and G. cuvier. Using both the annotated and unannotated reads, we were able to determine that the Carcharhinus galapagensis viromes were more similar to that of G. cuvier than they were to that of C. obscurus, suggesting that behavioral niche may be a more prominent driver of virome than host phylogeny.

The colocalizations of pulp neural stem cells markers with dentin matrix protein-1, dentin sialoprotein and dentin phosphoprotein in human denticle (pulp stone) lining cells.

BACKGROUND: The primary dentin, secondary dentin, and reactive tertiary dentin are formed by terminal differentiated odontoblasts, whereas atubular reparative tertiary dentin is formed by odontoblast-like cells. Odontoblast-like cells differentiate from pulpal stem cells, which express the neural stem cell markers nestin, S100ß, Sox10, and P0. The denticle (pulp stone) is an unique mineralized extracellular matrix that frequently occurs in association with the neurovascular structures in the dental pulp. However, to date, the cellular origin of denticles in human dental pulp is unclear. In addition, the non-collagenous extracellular dentin matrix proteins dentin matrix protein 1 (DMP1), dentin sialoprotein (DSP), and dentin phosphoprotein (DPP) have been well characterized in the dentin matrix, whereas their role in the formation and mineralization of the denticle matrix remains to be clarified. METHODS: To characterize the formation of denticle, healthy human third molars (n = 59) were completely sectioned and evaluated by HE staining in different layers at 720 µm intervals. From these samples, molars with (n = 5) and without denticles (n = 8) were selected. Using consecutive cryo-sections from a layer containing denticles of different sizes, we examined DMP1, DSP, and DPP in denticle lining cells and tested their co-localizations with the glial stem cell markers nestin, S100ß, Sox10, and P0 by quantitative and double staining methods. RESULTS: DMP1, DSP and DPP were found in odontoblasts, whereas denticle lining cells were positive only for DMP1 and DSP but not for DPP. Nestin was detected in both odontoblasts and denticle lining cells. S100ß, Sox10, and P0 were co-localized with DMP1 and DSP in different subpopulations of denticle lining cells. CONCLUSIONS: The co-localization of S100ß, Sox10, and P0 with DMP1 and DSP in denticle lining cells suggest that denticle lining cells are originated from glial and/or endoneurial mesenchymal stem cells which are involved in biomineralization of denticle matrix by secretion of DMP1 and DSP. Since denticles are atubular compared to primary, secondary, reactionary tertiary dentin and denticle formed by odontoblasts, our results suggest that DPP could be one of the proteins involved in the complex regulation of dentinal tubule formation.

Structural Changes and Mechanical Resistance of Claws and Denticles in Coconut Crabs of Different Sizes.

The exoskeleton of the pinching side of claws with denticles and of the outer side without them on the coconut crab, Birgus latro, which is a rare organism, were studied using a materials science approach. The mechanical resistance of three claws of different sizes was investigated along the exoskeleton thickness from the outer surface to the inner surface, and the results were compared, including the contribution of the microstructure and chemical compositions. Mechanical properties, hardness (H) and stiffness (Er), were probed through nanoindentation tests. The results showed the H, Er, microstructures, and chemical components of the exocuticle and endocuticle layers were almost the same, in a BW range of 300 g to 1650 g. At the same time, the H and Er near the surface of the denticles of a small coconut crab of 300 g were lower than those of other large coconut crabs. The microstructure of the denticles was clearly different from that of the exocuticle, but the maximum mechanical properties near their surface indicated almost the same values, Hmax = 4 GPa and Er(max) = 70 GPa, regardless of being on the pinching side or the outer side. A denticle can be regarded as a bulge of the cuticle without phosphorus and with high magnesium. The results provided novel information that expanded our knowledge about the claw microstructure of coconut crabs with different body sizes, and may be used in further studies.

Morfología microestructural dérmica en juveniles de dos especies hermanas de tiburón pala, Sphyrna tiburo y S. vespertina / Dermal microstructural morphology in juveniles of two sister species of bonnethead shark, Sphyrna tiburo and S. vespertina

RESUMEN: Los dentículos dérmicos son estructuras dermales presentes en el grupo de los condrictios, tienen un papel muy importante en su biología y se les ha utilizado como un carácter taxonómico que permiten reconocer grupos o especies. Por lo que en el presente trabajo se compara la morfología dermal de los juveniles de dos especies de tiburones pala, Sphyrna tiburo y S. vespertina, cuyo origen evolutivo está emparentado con el cierre del istmo centroaméricano. Para ello se obtuvieron muestras dermales (1 cm2) de tres regiones corporales y se procesaron para obtener imágenes de alta resolución por medio de Microscopia electrónica de barrido (MEB). Los dentículos de ambas especies tienen un patrón morfológico común, con variaciones en la longitud de las prolongaciones de las crestas, área libre y superposición de los dentículos, y grado de notoriedad de la ornamentación microestructural.

SUMMARY: The dermal denticles are dermal structures present in the group of chondrichthyans, they have a very important role in their biology and they have been used as a taxonomic character that allows to recognize groups or species. Therefore, in the present work, the dermal morphology of the juveniles of two species of shovel sharks, Sphyrna tiburo and S. vespertina, whose evolutionary origin is related to the closure of the Central American isthmus, is compared. For this, dermal samples (1 cm2) from three body regions were obtained and processed to obtain high resolution images by means of scanning electron microscopy (SEM). The denticles of both species have a common morphological pattern, with variations in the length of the ridge extensions, free area and overlapping of the denticles, and the degree of notoriety of the microstructural ornamentation.

Morphological and Histochemical Characterization of the Dermal Plates of Pleco (Hypostomus plecostomus).

Studying the dermal skeleton in fish is valuable for phylogenetic specification. The current study describes the detailed structure of the plecostomus dermal skeleton, including its morphogenesis and distribution in the skin. The denticles have a crown and a basal part and are embedded in bony depressions, to which they are attached by denticle ligaments. During denticle morphogenesis, denticle papillae formed from denticle precursor cells align in two cellular layers: an outer ameloblast precursor layer and an inner odontoblast precursor layer. The ameloblast precursors and odontoblast precursors differentiate and secrete enamel and dentine, respectively. We used different histochemical techniques, including Crossmon's trichrome staining, Weigert-Van Gieson staining, periodic acid-Schiff (PAS) staining, combined Alcian blue (AB; pH 2.5)/PAS staining, Weigert-Van Gieson staining, Mallory trichrome staining, and AB staining to distinguish the dentine and denticle ligaments. We used acridine orange to detect lysosome activity during denticle eruption. Transmission electron microscopy was used to detect the denticle ultrastructure, and scanning electron microscopy was used to detect the topographic distributions of different types of dermal tissues in different anatomical regions.

An unusual case of 177 pieces of mandibular compound odontoma: 10-year follow-up.

Odontomas are benign and the most common odontogenic tumors. They are classified as compound or complex odontomas according to their radiological and histological features. They have slow growth potential and compound odontoma is more common. Since they are generally asymptomatic they may reach in excessive sizes. In our case we would like to present a case with a large (177 denticles) compound odontoma in mandible.

Shark skin-inspired designs that improve aerodynamic performance.

There have been significant efforts recently aimed at improving the aerodynamic performance of aerofoils through the modification of their surfaces. Inspired by the drag-reducing properties of the tooth-like denticles that cover the skin of sharks, we describe here experimental and simulation-based investigations into the aerodynamic effects of novel denticle-inspired designs placed along the suction side of an aerofoil. Through parametric modelling to query a wide range of different designs, we discovered a set of denticle-inspired surface structures that achieve simultaneous drag reduction and lift generation on an aerofoil, resulting in lift-to-drag ratio improvements comparable to the best-reported for traditional low-profile vortex generators and even outperforming these existing designs at low angles of attack with improvements of up to 323%. Such behaviour is enabled by two concurrent mechanisms: (i) a separation bubble in the denticle's wake altering the flow pressure distribution of the aerofoil to enhance suction and (ii) streamwise vortices that replenish momentum loss in the boundary layer due to skin friction. Our findings not only open new avenues for improved aerodynamic design, but also provide new perspective on the role of the complex and potentially multifunctional morphology of shark denticles for increased swimming efficiency.

Trunk neural crest origin of dermal denticles in a cartilaginous fish.

Cartilaginous fishes (e.g., sharks and skates) possess a postcranial dermal skeleton consisting of tooth-like "denticles" embedded within their skin. As with teeth, the principal skeletal tissue of dermal denticles is dentine. In the head, cranial neural crest cells give rise to the dentine-producing cells (odontoblasts) of teeth. However, trunk neural crest cells are generally regarded as nonskeletogenic, and so the embryonic origin of trunk denticle odontoblasts remains unresolved. Here, we use expression of FoxD3 to pinpoint the specification and emigration of trunk neural crest cells in embryos of a cartilaginous fish, the little skate (Leucoraja erinacea). Using cell lineage tracing, we further demonstrate that trunk neural crest cells do, in fact, give rise to odontoblasts of trunk dermal denticles. These findings expand the repertoire of vertebrate trunk neural crest cell fates during normal development, highlight the likely primitive skeletogenic potential of this cell population, and point to a neural crest origin of dentine throughout the ancestral vertebrate dermal skeleton.

Proceedings of the 2017 National Toxicology Program Satellite Symposium.

The 2017 annual National Toxicology Program Satellite Symposium, entitled "Pathology Potpourri," was held in Montreal, Quebec, Canada at the Society of Toxicologic Pathology's 36th annual meeting. The goal of this symposium was to present and discuss challenging diagnostic pathology and/or nomenclature issues. This article presents summaries of the speakers' talks along with select images that were used by the audience for voting and discussion. Various lesions and other topics covered during the symposium included renal papillary degeneration in perinatally exposed animals, an atriocaval mesothelioma, an unusual presentation of an alveolar-bronchiolar carcinoma, a paraganglioma of the organ of Zuckerkandl (also called an extra-adrenal pheochromocytoma), the use of human muscle samples to illustrate the challenges of manual scoring of fluorescent staining, intertubular spermatocytic seminomas, medical device pathology assessment and discussion of the approval process, collagen-induced arthritis, incisor denticles, ameloblast degeneration and poorly mineralized enamel matrix, connective tissue paragangliomas, microcystin-LR toxicity, perivascular mast cells in the forebrain thalamus unrelated to treatment, and 2 cases that provided a review of the International Harmonization of Nomenclature and Diagnostic Criteria (INHAND) bone nomenclature and recommended application of the terminology in routine nonclinical toxicity studies.

SoxNeuro and Shavenbaby act cooperatively to shape denticles in the embryonic epidermis of Drosophila.

During development, extracellular signals are integrated by cells to induce the transcriptional circuitry that controls morphogenesis. In the fly epidermis, Wingless (Wg)/Wnt signaling directs cells to produce either a distinctly shaped denticle or no denticle, resulting in a segmental pattern of denticle belts separated by smooth, or 'naked', cuticle. Naked cuticle results from Wg repression of shavenbaby (svb), which encodes a transcription factor required for denticle construction. We have discovered that although the svb promoter responds differentially to altered Wg levels, Svb alone cannot produce the morphological diversity of denticles found in wild-type belts. Instead, a second Wg-responsive transcription factor, SoxNeuro (SoxN), cooperates with Svb to shape the denticles. Co-expressing ectopic SoxN with svb rescued diverse denticle morphologies. Conversely, removing SoxN activity eliminated the residual denticles found in svb mutant embryos. Furthermore, several known Svb target genes are also activated by SoxN, and we have discovered two novel target genes of SoxN that are expressed in denticle-producing cells and that are regulated independently of Svb. We conclude that proper denticle morphogenesis requires transcriptional regulation by both SoxN and Svb.

Developing an ancient epithelial appendage: FGF signalling regulates early tail denticle formation in sharks.

BACKGROUND: Vertebrate epithelial appendages constitute a diverse group of organs that includes integumentary structures such as reptilian scales, avian feathers and mammalian hair. Recent studies have provided new evidence for the homology of integumentary organ development throughout amniotes, despite their disparate final morphologies. These structures develop from conserved molecular signalling centres, known as epithelial placodes. It is not yet certain whether this homology extends beyond the integumentary organs of amniotes, as there is a lack of knowledge regarding their development in basal vertebrates. As the ancient sister lineage of bony vertebrates, extant chondrichthyans are well suited to testing the phylogenetic depth of this homology. Elasmobranchs (sharks, skates and rays) possess hard, mineralised epithelial appendages called odontodes, which include teeth and dermal denticles (placoid scales). Odontodes constitute some of the oldest known vertebrate integumentary appendages, predating the origin of gnathostomes. Here, we used an emerging model shark (Scyliorhinus canicula) to test the hypothesis that denticles are homologous to other placode-derived amniote integumentary organs. To examine the conservation of putative gene regulatory network (GRN) member function, we undertook small molecule inhibition of fibroblast growth factor (FGF) signalling during caudal denticle formation. RESULTS: We show that during early caudal denticle morphogenesis, the shark expresses homologues of conserved developmental gene families, known to comprise a core GRN for early placode morphogenesis in amniotes. This includes conserved expression of FGFs, sonic hedgehog (shh) and bone morphogenetic protein 4 (bmp4). Additionally, we reveal that denticle placodes possess columnar epithelial cells with a reduced rate of proliferation, a conserved characteristic of amniote skin appendage development. Small molecule inhibition of FGF signalling revealed placode development is FGF dependent, and inhibiting FGF activity resulted in downregulation of shh and bmp4 expression, consistent with the expectation from comparison to the amniote integumentary appendage GRN. CONCLUSION: Overall, these findings suggest the core GRN for building vertebrate integumentary epithelial appendages has been highly conserved over 450 million years. This provides evidence for the continuous, historical homology of epithelial appendage placodes throughout jawed vertebrates, from sharks to mammals. Epithelial placodes constitute the shared foundation upon which diverse vertebrate integumentary organs have evolved.

Morphology and distribution of taste papillae and oral denticles in the developing oropharyngeal cavity of the bamboo shark, Chiloscyllium punctatum.

Gustation in sharks is not well understood, especially within species that ingest food items using suction. This study examines the morphological and immunohistochemical characterisation of taste papillae and oral denticles in the oropharynx of the brown-banded bamboo shark Chiloscyllium punctatum and compares their distribution during development. Taste papillae of C. punctatum are located throughout the oropharyngeal region and are most concentrated on the oral valves (2125-3483 per cm2 in embryos; 89-111 per cm2 in mature adults) close to the tooth territories. Papillae appearance is comparable at all stages of development, with the exception of the embryos (unhatched specimens), where no microvilli are present. Oral valve papillae are comparable in structure to Type I taste buds of teleost fishes, whereas those of the rest of the oropharyngeal region are comparable to Type II. Both types of papillae show immunofluorescence for a number of markers of taste buds, including ß-Catenin and Sox2. Taste papillae densities are highest in embryos with 420-941 per cm2 compared to 8-29 per cm2 in mature adults. The total number of papillae remains around 1900 for all stages of development. However, the papillae increase in diameter from 72±1 µm (mean±s.e.m.) in embryos to 310±7 µm in mature individuals. Microvilli protrude in multiple patches at the apical tip of the papilla covering ∼0.5% of the papillar surface area. We further document the relationship between taste papillae and the closely associated oral denticles within the shark orophayngeal cavity. Oral denticles first break through the epithelium in the antero-central region of the dorsal oral cavity, shortly after the emergence of teeth, around time of hatching. Denticles are located throughout the oropharyngeal epithelium of both immature and mature stages, with the highest concentrations in the antero-dorsal oral cavity and the central regions of the pharynx. These denticle-rich areas of the mouth and pharynx are therefore thought to protect the epithelium, and importantly the taste papillae, from abrasion since they correlate with regions where potential food items are processed or masticated for consumption. Taste papillae and denticles are more dense in anterior oropharyngeal regions in close association with the oral jaws and teeth, and in the juvenile or hatchling shark taste units are functional, and innervated, allowing the shark to seek out food in utero, at birth or on emergence from the egg case.

Denticle-embedded ampullary organs in a Cretaceous shark provide unique insight into the evolution of elasmobranch electroreceptors.

Here, we report a novel type of dermal denticle (or placoid scale), unknown among both living and fossil chondrichthyan fishes, in a Cretaceous lamniform shark. By their morphology and location, these dermal denticles, grouped into clusters in the cephalic region, appear to have been directly associated with the electrosensory ampullary system. These denticles have a relatively enlarged (∼350 µm in diameter), ornamented crown with a small (∼100 µm) asterisk- or cross-shaped central perforation connected to a multi-alveolate internal cavity. The formation of such a complex structure can be explained by the annular coalescence and fusion, around an ampullary vesicle, of several developmental units still at papillary stage (i.e. before mineralization), leading to a single denticle embedding an alveolar ampulla devoid of canal. This differs from larger typical ampullae of Lorenzini with a well-developed canal opening in a pore of the skin and may represent another adaptive response to low skin resistance. Since it has been recently demonstrated that ampullary organs arise from lateral line placodes in chondrichthyans, this highly specialized type of dermal denticle (most likely non-deciduous) may be derived from the modified placoid scales covering the superficial neuromasts (pit organs) of the mechanosensory lateral line system of many modern sharks.

Biomimetic shark skin: design, fabrication and hydrodynamic function.

Although the functional properties of shark skin have been of considerable interest to both biologists and engineers because of the complex hydrodynamic effects of surface roughness, no study to date has successfully fabricated a flexible biomimetic shark skin that allows detailed study of hydrodynamic function. We present the first study of the design, fabrication and hydrodynamic testing of a synthetic, flexible, shark skin membrane. A three-dimensional (3D) model of shark skin denticles was constructed using micro-CT imaging of the skin of the shortfin mako (Isurus oxyrinchus). Using 3D printing, thousands of rigid synthetic shark denticles were placed on flexible membranes in a controlled, linear-arrayed pattern. This flexible 3D printed shark skin model was then tested in water using a robotic flapping device that allowed us to either hold the models in a stationary position or move them dynamically at their self-propelled swimming speed. Compared with a smooth control model without denticles, the 3D printed shark skin showed increased swimming speed with reduced energy consumption under certain motion programs. For example, at a heave frequency of 1.5 Hz and an amplitude of ± 1 cm, swimming speed increased by 6.6% and the energy cost-of-transport was reduced by 5.9%. In addition, a leading-edge vortex with greater vorticity than the smooth control was generated by the 3D printed shark skin, which may explain the increased swimming speeds. The ability to fabricate synthetic biomimetic shark skin opens up a wide array of possible manipulations of surface roughness parameters, and the ability to examine the hydrodynamic consequences of diverse skin denticle shapes present in different shark species.

Drosophila Dachsous and Fat polarize actin-based protrusions over a restricted domain of the embryonic denticle field.

Atypical cadherins Dachsous (Ds) and Fat coordinate the establishment of planar polarity, essential for the patterning of complex tissues and organs. The precise mechanisms by which this system acts, particularly in cases where Ds and Fat act independently of the 'core' frizzled system, are still the subject of investigation. Examining the deployment of the Ds-Fat system in different tissues of the model organism Drosophila, has provided insights into the general mechanisms by which polarity is established and propagated to coordinate outcomes across a field of cells. The Drosophila embryonic epidermis provides a simple model epithelia where the establishment of polarity can be observed from start to finish, and in the absence of proliferation, over a fixed number of cells. Using the asymmetric placement of f-actin during denticle assembly as a read-out of polarity, we examine the requirement for Ds and Fat in establishing polarity across the denticle field. Comparing detailed phenotypic analysis with steady state protein enrichment revealed a spatially restricted requirement for the Ds-Fat system within the posterior denticle field. Ectopic Ds signaling provides evidence for a model whereby Ds acts to asymmetrically enrich Fat in a neighboring cell, in turn polarizing the cell to specify the position of the actin-based protrusions at the cell cortex.

Histochemistry and functional organization of the dorsal skin of Ancistrus dolichopterus (Siluriformes: Loricariidae)

The structural organization and histo-cytochemical features of dorsal skin of Ancistrus dolichopterus (acari bodo) are the main focus of this work. The epidermis, dermis and subcutis are the principal layers of the skin. The epidermis mainly consists of epithelial and mucous cells. Interspersed between them are lymphocytes, pigment cells, eosinophilic granular cells (EGC), and the taste buds as sensory structures. The high number of EGCs is implicated in general and specific immunological defense from pathogenic bacteria and multicellular parasites. The epithelial cells and mucous cells contain glycoproteins with oxidizable vicinal diols, carboxyl groups and O-sulphate esters and their high secretory activity is correlated with the bottom dwelling habit of this species. A thick stratum laxum contains overlapping osteoderms bearing denticles, and the stratum compactum make the integument thicker to help the fish in negative buoyancy for maneuvering near the bottom and protection. The entire body surface is covered by conical, backwardly directed denticles. These are composed of a dentine cone, surrounding a pulp cavity with the top covered by mineralized cap, and are the true homologues of teeth. These structures provide effective protection from abrasion and enemies. These structural peculiarities and histochemical features indicate additional physiological role of the skin of A. dolichopterus.

A organização estrutural e aspectos histo-citiquímicos da pele dorsal de Ancistrus dolichopterus (acari-bodó) são os principais alvos do presente estudo. A epiderme, a derme e a hipoderme são as principais camadas da pele. A epiderme consiste principalmente de células epiteliais e mucosas. Intercalados entre elas estão os linfócitos, as células pigmentares, as células granulares eosinofílicas (CGE), e as papilas gustativas como estruturas sensoriais. Um grande número de CGEs está relacionado em geral com a defesa imunológica específica de bactérias patogênicas e parasitas multicelulares. As células epiteliais e as células mucosas contem glicoproteínas com grupos diol oxidáveis, grupos carboxilas e ésteres O-sulfatados sendo que sua alta atividade secretória está correlacionada com o comportamento bentônico, de fundo, dessa espécie. Um espesso stratum laxum contém osteodermos sobrepostos, parecidos com dentículos, e o stratum compactum que torna o tegumento mais espesso, contribuindo com a flutuação negativa necessária ao movimento perto do fundo e proteção. Toda a superfície do corpo é coberta por dentículos cônicos retrodirecionados. Esses dentículos são compostos por um cone de dentina, envolvendo uma cavidade pulpar e com o ápice coberto por uma capa mineralizada, verdadeiros homólogos dos dentes. Essas estruturas oferecem efetiva proteção contra abrasão e oponentes. Essas peculiaridades estruturais e aspectos histoquímicos sugerem a existência de uma função fisiológica adicional para a pele de A. dolichopterus.