Microstructural and compositional variation in pacu and piranha teeth related to diet specialization (Teleostei: Serrasalmidae).

In any vertebrate group, tooth shape is known to fit with a biological function related to diet. However, little is known about the relationships between diet and tooth microstructure and composition in teleost fishes. In this work, we describe the external morphology, internal microstructure and elemental composition of the oral teeth of three representative species of the family Serrasalmidae having different feeding habits (herbivorous vs. omnivorous vs. carnivorous). We used backscattered-electron imaging and low vacuum environmental scanning electron microscope to compare the organization and mineralization of tooth layers as well as energy dispersive X-ray microanalysis and Raman microspectrometry to investigate the elemental composition, Ca/P ratio and mineralogy of the most superficial layers. Oral teeth of each serrasalmid species have the same internal organization based on five distinctive layers (i.e. pulp, dentine, inner enameloid, outer enameloid and cuticle) but the general tooth morphology is different according to diet. Microstructural and compositional variation of the cuticle and iron-enrichment of superficial layers were highlighted between herbivorous and carnivorous species. Iron is more concentrated in teeth of the herbivorous species where it is associated with a thicker cuticle explaining the more intense red-pigmentation of the cutting edges of oral teeth. The iron-enrichment is interpreted as a substitution of Ca by Fe in the hydroxyapatite. These traits are discussed in the light of the evolutionary history of the family. Further considerations and hypotheses about the formation and origin of the mineralized tooth layers and especially the iron-rich superficial layers in teleost fishes are suggested.

Biodegradable Cell Microcarriers Based on Chitosan/Polyester Graft-Copolymers.

Self-stabilizing biodegradable microcarriers were produced via an oil/water solvent evaporation technique using amphiphilic chitosan-g-polyester copolymers as a core material in oil phase without the addition of any emulsifier in aqueous phase. The total yield of the copolymer-based microparticles reached up to 79 wt. %, which is comparable to a yield achievable using traditional emulsifiers. The kinetics of microparticle self-stabilization, monitored during their process, were correlated to the migration of hydrophilic copolymer's moieties to the oil/water interface. With a favorable surface/volume ratio and the presence of bioadhesive natural fragments anchored to their surface, the performance of these novel microcarriers has been highlighted by evaluating cell morphology and proliferation within a week of cell cultivation in vitro.

When Microplastic Is Not Plastic: The Ingestion of Artificial Cellulose Fibers by Macrofauna Living in Seagrass Macrophytodetritus.

Dead leaves of the Neptune grass, Posidonia oceanica (L.) Delile, in the Mediterranean coastal zone, are colonized by an abundant "detritivorous" invertebrate community that is heavily predated by fishes. This community was sampled in August 2011, November 2011, and March 2012 at two different sites in the Calvi Bay (Corsica). Ingested artificial fibers (AFs) of various sizes and colors were found in 27.6% of the digestive tracts of the nine dominant species regardless of their trophic level or taxon. No seasonal, spatial, size, or species-specific significant differences were revealed; suggesting that invertebrates ingest AFs at constant rates. Results showed that, in the gut contents of invertebrates, varying by trophic level, and across trophic levels, the overall ingestion of AFs was low (approximately 1 fiber per organism). Raman spectroscopy revealed that the ingested AFs were composed of viscose, an artificial, cellulose-based polymer. Most of these AFs also appeared to have been colored by industrial dyes. Two dyes were identified: Direct Blue 22 and Direct Red 28. The latter is known for being carcinogenic for vertebrates, potentially causing environmental problems for the P. oceanica litter community. Techniques such as Raman spectroscopy are necessary to investigate the particles composition, instead of relying on fragment size or color to identify the particles ingested by animals.

New insights into the role of the pharyngeal jaw apparatus in the sound-producing mechanism of Haemulon flavolineatum (Haemulidae).

Grunts are fish that are well known to vocalize, but how they produce their grunting sounds has not been clearly identified. In addition to characterizing acoustic signals and hearing in the French grunt Haemulon flavolineatum, the present study investigates the sound-production mechanism of this species by means of high-speed X-ray videos and scanning electron microscopy of the pharyngeal jaw apparatus. Vocalizations consist of a series of stridulatory sounds: grunts lasting ~47 ms with a mean period of 155 ms and a dominant frequency of ~700 Hz. Auditory capacity was determined to range from 100 to 600 Hz, with greatest sensitivity at 300 Hz (105.0±11.8 dB re. 1 µPa). This suggests that hearing is not tuned exclusively to detect the sounds of conspecifics. High-speed X-ray videos revealed how pharyngeal jaws move during sound production. Traces of erosion on teeth in the fourth ceratobranchial arch suggest that they are also involved in sound production. The similarity of motor patterns of the upper and lower pharyngeal jaws between food processing and sound production indicates that calling is an exaptation of the food-processing mechanism.

An Empirical Model Linking Physico-Chemical Biomaterial Characteristics to Intra-Oral Bone Formation.

Facial trauma, bone resection due to cancer, periodontal diseases, and bone atrophy following tooth extraction often leads to alveolar bone defects that require bone regeneration in order to restore dental function. Guided bone regeneration using synthetic biomaterials has been suggested as an alternative approach to autologous bone grafts. The efficiency of bone substitute materials seems to be influenced by their physico-chemical characteristics; however, the debate is still ongoing on what constitutes optimal biomaterial characteristics. The purpose of this study was to develop an empirical model allowing the assessment of the bone regeneration potential of new biomaterials on the basis of their physico-chemical characteristics, potentially giving directions for the design of a new generation of dental biomaterials. A quantitative data set was built composed of physico-chemical characteristics of seven commercially available intra-oral bone biomaterials and their in vivo response. This empirical model allowed the identification of the construct parameters driving optimized bone formation. The presented model provides a better understanding of the influence of driving biomaterial properties in the bone healing process and can be used as a tool to design bone biomaterials with a more controlled and custom-made composition and structure, thereby facilitating and improving the clinical translation.

Effect of lab procedures and ultrasonic bath cleaning on customized implant abutment pollution: An in vitro study.

AIM: The aim of this study was to validate the effectiveness of a simplified ultrasonic cleaning protocol to clean customized abutments. The second purpose was to investigate the percentage of pollutants on customized abutments delivered by the implants company and the additional effect of dental laboratory manipulations. MATERIALS AND METHODS: Twenty-four customized abutments were divided in two groups, 12 returning from the implant company and 12 others returning from the dental laboratory. In each group, there were 6 zirconia (Zr02) abutments and 6 titanium (Ti) abutments. For each conditions, half of the samples were clean with the experimental protocol and the other were left as delivered by the company. The two steps cleaning protocol consisted of mechanical treatment with a sterile compress soaked in a detergent over the transgingival part of the abutment followed by 3 successive ultrasound baths for 2 min/bath. The presence of pollutants was quantified by scanning electron microscopy. RESULTS: The suggested cleaning method allowed to significantly decrease the quantity of pollutants (p=0.0006). The abutments returning from the dental laboratory were significantly more polluted than those coming directly from the implant company (p=0.0043). The cleaning effect was highly significant in both groups (p<0.0001). The quantity of pollutants before cleaning were similar in the titanium and in the zirconia groups and the cleaning effect was highly significant in both groups (p=0.0009). CONCLUSION: The tested cleaning protocol was successful on the customized abutments from each group.

Ultrastructure of sonic muscles of piranhas (Serrasalmidae).

Among piranhas, different species are able to produce sounds but not all of them use the same mechanism. In all species, the sound-producing muscle originates on the second vertebra, but the insertion differs. Pygopristis denticulata can produce two kinds of pulsed sounds emitted in trains. Its sound production mechanism is mainly based on a muscle bundle that inserts between the two first ribs. In Catoprion mento, the anterior part of the sonic muscle inserts directly on the swim bladder. The most derived species (Serrasalmus and Pygocentrus) make all harmonic tonal sounds. Their sonic muscles constitute a single functional unit transversally surrounding the swim bladder. This study aims to study the ultrastructure of sonic muscles in nine species from these four genera. Epaxial muscles were compared with sonic muscles, and the sonic muscles of the different species were compared between them. Results confirmed ultrastructure modifications in the sonic muscles in comparison to epaxial muscles. Fibers of the sonic muscle are thinner and possess a thicker subsarcolemmal ring housing mitochondria. In sonic muscles, myofibrils are also proportionally less abundant, and their sarcomeres are longer and thinner. Some of these differences allows to separate basal species (e.g., P. denticulata) from more derived species (genera Pygocentrus and Serrasalmus) and supposedly support the observed differences in the acoustic abilities.

Astin C Production by the Endophytic Fungus Cyanodermella asteris in Planktonic and Immobilized Culture Conditions.

The fungal endophyte Cyanodermella asteris (C. asteris) has been recently isolated from the medicinal plant Aster tataricus (A. tataricus). This fungus produces astin C, a cyclic pentapeptide with anticancer and anti-inflammatory properties. The production of this secondary metabolite is compared in immobilized and planktonic conditions. For immobilized cultures, a stainless steel packing immersed in the culture broth is used as a support. In these conditions, the fungus exclusively grows on the packing, which provides a considerable advantage for astin C recovery and purification. C. asteris metabolism is different according to the culture conditions in terms of substrate consumption rate, cell growth, and astin C production. Immobilized-cell cultures yield a 30% increase of astin C production, associated with a 39% increase in biomass. The inoculum type as spores rather than hyphae, and a pre-inoculation washing procedure with sodium hydroxide, turns out to be beneficial both for astin C production and fungus development onto the support. Finally, the influence of culture parameters such as pH and medium composition on astin C production is evaluated. With optimized culture conditions, astin C yield is further improved reaching a five times higher final specific yield compared to the value reported with astin C extraction from A. tataricus (0.89 mg g-1 and 0.16 mg g-1 respectively).

Microplastics in livers of European anchovies (Engraulis encrasicolus, L.).

Microplastics (MPs) are thought to be ingested by a wide range of marine organisms before being excreted. However, several studies in marine organisms from different taxa have shown that MPs and nanoplastics could be translocated in other organs. In this study, we investigated the presence of MPs in the livers of commercial zooplanktivorous fishes collected in the field. The study focuses mainly on the European anchovy Engraulis encrasicolus but concerns also the European pilchard Sardina pilchardus and the Atlantic herring Clupea harengus. Two complementary methodologies were used to attest the occurrence of MPs in the hepatic tissue and to exclude contamination. 1) MPs were isolated by degradation of the hepatic tissue. 2) Cryosections were made on the livers and observed in polarized light microscopy. Both methods separately revealed that MPs, mainly polyethylene (PE), were translocated into the livers of the three clupeid species. In anchovy, 80 per cent of livers contained relatively large MPs that ranged from 124 µm to 438 µm, showing a high level of contamination. Two translocation pathways are hypothesized: (i) large particles found in the liver resulted from the agglomeration of smaller pieces, and/or (ii) they simply pass through the intestinal barrier. Further studies are however required to understand the exact process.

Morphology of the filtration apparatus of three planktivorous fishes and relation with ingested anthropogenic particles.

Anthropogenic particles (APs), including microplastics, are ingested by a wide variety of marine organisms. Exposure of Clupeiformes (e.g. herrings, anchovies, sardines) is poorly studied despite their economic and ecological importance. This study aims to describe the morphology of the filtration apparatus of three wild-caught Clupeiformes (Sardina pilchardus, Clupea harengus and Engraulis encrasicolus) and to relate the results to ingested APs. Consequently, the species with the more efficient filtration apparatus will be more likely to ingest APs. We hypothesized that sardines were the most exposed species. The filtration area and particle retention threshold were determined in the three species, with sardines displaying the highest filtration area and the closest gill rakers. Sardines ingested more fibers and smaller fragments, confirming that it is the most efficient filtering species. These two results lead to the conclusion that, among the three studied, the sardine is the species most exposed to APs.

Histopathological effects of Aspergillus clavatus (Ascomycota: Trichocomaceae) on larvae of the southern house mosquito, Culex quinquefasciatus (Diptera: Culicidae).

Aspergillus clavatus (Ascomycota: Trichocomaceae) was previously found to be an opportunistic pathogen of mosquitoes (Diptera: Culicidae). In the present study, the mechanism leading to its insecticidal activity was investigated regarding histological damages on Culex quinquefasciatus larvae exposed to A. clavatus spores. Multiple concentration assays using spore suspensions (0.5-2.5 × 10(8) spores ml(-1)) revealed 17.0-74.3 % corrected mortalities after 48 h exposure. Heat-deactivated spores induced a lower mortality compared to nonheated spores suggesting that insecticidal effects are actively exerted. Spore-treated and untreated larvae were prepared for light microscopy as well as for scanning and transmission electron microscopy. Spores failed to adhere to the external body surface (except the mouth parts) of these aquatic immature stages but progressively filled the digestive tract where their metabolism seemed to activate. In parallel, the internal tissues of the larvae, i.e. the midgut wall, the skeletal muscles, and the cuticle-secreting epidermis, were progressively destroyed between 8 and 24 h of exposure. These observations suggest that toxins secreted by active germinating spores of A. clavatus in the digestive tract altered the larval tissues, leading to their necrosis and causing larval death. Fungal proliferation and sporulation then occurred during a saprophytic phase. A. clavatus enzymes or toxins responsible for these pathogenic effects need to be identified in further studies before any use of this fungus in mosquito control.

Dentinal tubules sealing by means of diode lasers (810 and 980 nm): a preliminary in vitro study.

OBJECTIVE: The aim of this study was to evaluate the effect on dentinal surfaces of diode lasers (810 and 980 nm) at different parameters. MATERIALS AND METHODS: Twenty-four caries-free human impacted wisdom teeth were used. The crowns were sectioned transversely in order to expose the dentin. The smear layer was removed by a 1 min application of ethylenediaminetetraacetic acid (EDTA). Each surface was divided into four quadrants irradiated at a different output power setting for each kind of laser: 0.8, 1, 1.6, and 2 W (energy densities: 2547, 3184, 5092, and 6366 J/cm(2), irradiation speed 1 mm/sec; optical fiber diameter: 200 µm; continuous and noncontact mode). Half of the samples were stained with a graphite paste. All specimens were sent for scanning electron microscopic (SEM) analysis. Pulp temperature increases in additional 20 teeth were measured by a thermocouple. RESULTS: Diode laser irradiations at 0.8 and 1 W led to occlusion or narrowing of dentin tubules without provoking fissures or cracks. The application of graphite paste increased the thermal effects in dentin. Measurements of pulp temperature showed that irradiations at 0.8 and 1 W for a period of 10 sec in continuous mode increased pulp temperature (T ≤2°C). CONCLUSIONS: Diode lasers (810 and 980 nm) used at 0.8 and 1 W for 10 sec in continuous mode were able to seal the dentin tubules. These parameters can be considered harmless for pulp vitality, and may be effective in the treatment of dentinal hypersensitivity.

Influence of sodium hypochlorite on Er:YAG laser-irradiated dentin and its effect on the quality of adaptation of the composite restoration margins.

OBJECTIVE: The aims of this in vitro study were to evaluate: (1) the influence of 5% NaOCl application on Er:YAG-irradiated dentin; and (2) its effect on the quality of adaptation of the composite restoration margins. BACKGROUND DATA: Previous research has shown that Er:YAG dentin irradiation produces a thermally affected tissue layer that results in lower bond strength than that of nonirradiated dentin. The removal of this thermally-affected layer may enhance the quality of dentin bonding. MATERIALS AND METHODS: Forty-nine caries-free extracted human molars were transversely sectioned in order to totally expose the dentin. Four standardized cavities were created on the dentinal surface of each molar. First, two cavities were irradiated with Er:YAG laser (2.94 nm): 150 mJ, 10 Hz, variable square pulse (VSP) mode (100 µsec), beam diameter=0.9 mm, speed of irradiation=1 mm/sec, 20% air and 20% water. Then, one of irradiated cavities and one of nonirradiated cavities were treated for 30 sec with 5% NaOCl solution. Finally, they went through a standard bonding treatment for composite restoration, etching, bonding, and composite filling. We obtained four groups of cavities: (1) one control group of nonirradiated cavities not pretreated with NaOCl; (2) one group of nonirradiated cavities, pretreated with NaOCl; (3) one group of irradiated cavities, not pretreated with NaOCl; and (4) one group of irradiated cavities, pretreated with NaOCl. All samples were subjected to thermocycling. Every cavity was immersed into a 0.5% solution of methylene blue. The percentage of dye penetration (microleakage) in the composite-dentin interface was evaluated. Six molars were analyzed by scanning electron microscope. RESULTS: Dye infiltration depth was significantly reduced in irradiated cavities treated with 5% NaOCl solution. CONCLUSIONS: The application of a 5% NaOCl solution on Er:YAG irradiated cavities can significantly improve the marginal quality of composite bonding.

Polyolefin matrixes with permanent antibacterial activity: preparation, antibacterial activity, and action mode of the active species.

Poly[2-(tert-butylamino)ethyl methacrylate] (PTBAEMA) belongs to a novel class of water-insoluble biocides. Dispersion of a poly(ethylene-co-butylene)-b-poly[2-(tert-butylamino)ethyl methacrylate] diblock copolymer (PEB-b-PTBAEMA) within low-density polyethylene (LDPE) imparts antimicrobial properties to the polyolefin as assessed by the viable cell counting method against Escherichia coli (E. coli). This diblock copolymer has been synthesized by atom transfer radical polymerization (ATRP) with a poly(ethylene-co-butylene) (PEB) oligomer end-capped by an activated bromide as a macroinitiator for the polymerization of 2-(tert-butylamino)ethyl methacrylate (TBAEMA). Morphological changes of E. coli bacteria in contact with modified LDPE have been observed by transmission and scanning electron microscopy and indicate that the diblock copolymer is bactericide rather than bacteriostatic. Finally, the action mode of the PEB-b-PTBAEMA copolymer more likely relies on the displacement of the Ca(2+) and/or Mg(2+) ions of the outer membrane of the bacteria, which is disorganized and finally disrupted.