Cutaneous myxosarcoma in a Syrian hamster (Mesocricetus auratus)

The pathological findings in a 2-years-old Syrian hamster (Mesocricetus auratus) with a cutaneous myxosarcoma are described. Grossly, there was a large cutaneous mass in the right cervical region. Microscopical evaluation revealed a myxosarcoma characterized by pleomorphic, fusiform cells loosely arranged, randomly distributed, and presenting a moderate amount of basophilic amorphous stroma. There were hemorrhagic areas within the tumor. The basophilic amorphous stroma was positive to Alcian blue confirming the presence of a mucopolysaccharide matrix. Immunohistochemically, the neoplastic cells expressed vimentin, and were negative for cytokeratin or glial fibrillary acidic protein (GFAP).

Effect of pH variations in a cycling model on the properties of restorative materials

This study evaluated the effect of cycling various pH demineralizing solutions on the surface hardness, fluoride release and surface properties of restorative materials (Ketac-Fil Plus, Vitremer, Fuji II LC, Freedom and Fluorofil). Thirty specimens of each material were made and the surface hardness measured. The specimens were randomized into five groups according to the pH (4.3; 4.6; 5.0; 5.5 and 6.2) of the demineralizing solution. The specimens were submitted to pH-cycling for 15 days. The specimens remained in the demineralizing solution for six hours and in the remineralizing solution for 18 hours. Then, the surface hardness (SH) was remeasured and the surface properties were assessed. Fluoride release was determined daily. Data from SH and the percentage of alteration in surface hardness were analyzed by analysis of variance (p < 0.05); the Kruskal-Wallis test was performed for the fluoride release results. When hardness was compared, the variation in pH led to a positive correlation for glass ionomer cements and a negative correlation for fluoride release. For polyacid-modified resin composites, a negative correlation was found with regards to fluoride release; no significant correlation was observed for hardness. Surface properties were influenced: an acidic pH led to a greater alteration, except for polyacid-modified resin composites. The pH of the demineralizing solution influenced fluoride release from the tested materials. The pH variation altered hardness and surface properties of glass ionomer cements but did not influence polyacid-modified resin composites.

Stability of cp-Ti and Ti-6Al-4V alloy for dental implants as a function of saliva pH - an electrochemical study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

In Vitro Evaluation of Surface Roughness and Microhardness of Restorative Materials Submitted to Erosive Challenges

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of ion supplementation of a commercial soft drink on tooth enamel erosion

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of Nd:YAG Irradiation and Fluoride Application on Dentine Resistance to Erosion in Vitro

Objective: In this paper we evaluated the effect of two fluoridated agents and Nd:YAG irradiation separately and in combination on dentine resistance to erosion. Background Data: The morphological changes in dentin induced by laser treatment may reduce the progression of erosive lesions. Due to the possibility of a synergistic effect of laser with fluoride, this study was conducted. Materials and Methods: Eighty bovine dentine samples (4 x 4 mm) were randomly divided into eight groups, according to the following treatments: G1: untreated (control); G2: acidic phosphate fluoride gel (APF 1.23%) for 4 min; G3: fluoride varnish (NaF 2.26%) for 6 h; G4: 0.5 W Nd: YAG laser (250 mu sec pulse, 10 Hz, 35 J/cm(2), 30 sec); G5: 0.75 W Nd: YAG laser (52.5 J/cm(2)); G6: 1.0 W Nd: YAG laser (70 J/cm(2)); G7: APF + 0.75 W Nd: YAG laser; and G8: NaF + 0.75 W Nd: YAG laser. After the treatments, half of each dentine surface was protected with nail varnish. The samples were stored in artificial saliva (30 mL/sample) for 24 h and submitted to four erosive 1-min cycles. Between the erosive attacks, the blocks were maintained in artificial saliva for 59 min. The erosive wear was evaluated by profilometry. Results: The mean wear (+/- SD, mu m) was: G1: 1.20 +/- 0.20; G2: 0.47 +/- 0.06; G3: 0.81 +/- 0.11; G4: 1.47 +/- 0.32; G5: 1.52 +/- 0.24; G6: 1.49 +/- 0.30; G7: 0.49 +/- 0.11; and G8: 1.06 +/- 0.31 (Tukey's test, p < 0.05). Conclusions: Laser irradiation was not able to reduce dentine erosion. However, fluoride application was able to increase the dentine's resistance to erosion, and APF showed better results than fluoride varnish.

Current status of pulp capping with dentin adhesive systems: a review

Several studies have assessed the morphology and thickness of hybrid layer, the dentin bend strengths as well as sealing ability of dentin adhesive systems. However, few in vivo studies have evaluated the biocompatibility of the adhesive systems following application to deep dentin or directly to the pulp of human teeth. Many studies performed in non-human primate teeth or teeth of rats have reported pulp healing and dentin bridging following pulp capping with bonding agents. In addition, a few clinical and radiographical reports of the success of resin pulp capping have been described in the dental literature.Objectives: the aim of this review was to evaluate the literature on pulp responses following total acid etching and application of adhesive resins on deep cavities or pulp exposures. In addition, the clinical/radiographical evidence for the apparent success of vital pulp therapy and results obtained from animal and human studies were compared and discussed.Significance and conclusions: the self-etching adhesive systems may be useful and safe when applied on dentin, In contrast, persistent inflammatory reactions as well as delay in pulpal healing and failure of dentin bridging were seen in human pulps capped with bonding agents. The results observed in animal teeth cannot be directly extrapolated to human clinical conditions. Consequently, vital pulp therapy using acidic agents and adhesive resins seems to be contraindicated. (C) 2000 Academy of Dental Materials. Published by Elsevier B.V. Ltd. All rights reserved.

Response of human pulps following acid conditioning and application of a bonding agent in deep cavities

Objectives. The aim of this in vivo study was to evaluate the human dental pulp response when a one-bottle adhesive system was applied on etched or unetched deep dentine.Methods. Eighteen class V deep cavity preparations were divided in three groups: group 1-total etching + two coats of single bond (SB) + composite resin (Z-100); group 2-enamel etching + two coats of SB + Z-100, group 3-cavity floor lined with a calcium hydroxide liner (Dycal) + acid-etching of enamel and lateral walls + two coats of SB + Z-100. Two teeth were used as intact control group. After 30 days the teeth were extracted and processed through H and E, Masson's trichrome and Brown and Brenn staining techniques.Results. Moderate inflammatory response, disorganization of pulp tissue, as well as, deposition of thin layer of reactionary dentin were observed in group 1 teeth in which the remaining dentin thickness (RDT) was less than 300 mum. These histological findings appear to be related to long resin tags formation and bonding agent diffusion through dentinal tubules. In group 2, slight inflammatory response was observed only in one tooth in which the RDT was 162 mum. In group 3, all the teeth showed normal histological characteristics which were similar to the intact control group. Presence of bacteria was not correlated with the intensity of pulpal response. The patients reported no symptoms during the experiment. Radiographic evaluation showed no periapical pathology for any of the teeth,Significance and conclusions. Acid-etched deep dentin (RDT less than 300 mum) lined with SB causes more intense pulpal response than unetched deep dentin. Based on the results observed in the present study and the conditions in which it was carried out, we recommend the application of a biocompatible liner before etching deep dentin and applying SB. (C) 2002 Academy of Dental Materials. Published by Elsevier B.V. Ltd. All rights reserved.

Evaluation of pH, available chlorine content, and antibacterial activity of endodontic irrigants and their combinations against Enterococcus faecalis

Objectives. The objectives of this study were to evaluate pH, available chlorine content, and antibacterial activity of endodontic irrigants and their combinations.Study design. The pH and chlorine content of sodium hypochlorite (NaOCl) were analyzed pure and in combination with 10% citric acid (CA) and apple vinegar (AV). The antibacterial effect of the following solutions was measured by direct contact test against Enterococcus faecalis: 2.5% NaOCl, 2.5% NaOCl + 10% CA (7:3), 2.5% NaOCl + AV (5:5), 10% CA, and AV. Sterile saline was used as control. The colony-forming units were determined by serial decimal dilutions.Results. The combination of 2.5% NaOCl with CA or AV lowered the pH and the chlorine content. NaOCl, alone or in combination was able to eliminate E. faecalis in 30 seconds, and CA, after 10 minutes. AV promoted reduction (32.2%) after 10 minutes.Conclusions. NaOCl with acidic solutions lowered the pH and the chlorine content, but did not alter its antibacterial effect. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011; 112:132-135)

Comparative scanning electron microscopic study of the effect of different dental conditioners on dentin micromorphology

This study evaluated comparatively by scanning electron microscopy (SEM) the effect of different dental conditioners on dentin micromorphology, when used according to the same protocol. Forty dentin sticks were obtained from 20 caries-free third human molars and were assigned to 4 groups corresponding to 3 conditioners (phosphoric acid 37%, Clearfil SE Bond and iBond) and an untreated control group. After application of the conditioners, the specimens were immersed in 50% ethanol solution during 10 s, chemically fixed and dehydrated to prepare them to SEM analysis. In the control group, dentin surface was completely covered by smear layer and all dentinal tubules were occluded. In the phosphoric acid-etched group, dentin surface was completely clean and presented exposed dentinal tubule openings; this was the only group in which the tubules exhibited the funnel-shaped aspect. In the groups conditioned with Clearfil SE Bond primer and iBond, which are less acidic than phosphoric acid, tubule openings were occluded or partially occluded, though smear layer removal was observed. SE Bond was more efficient in removing the smear layer than iBond. In the Clearfil SE Bond group, the cuff-like aspect of peritubular dentin was more evident. It may be concluded all tested conditioners were able to change dentin morphology. However, it cannot be stated that the agent aggressiveness was the only cause of the micromorphological alterations because a single morphological pattern was not established for each group, but rather an association of different aspects, according to the aggressiveness of the tested conditioner.

Analysis of Bothrops jararacussu venomous gland transcriptome focusing on structural and functional aspects: I - gene expression profile of highly expressed phospholipases A(2)

Snake venom glands are a rich source of bioactive molecules such as peptides, proteins and enzymes that show important pharmacological activity leading to in local and systemic effects as pain, edema, bleeding and muscle necrosis. Most studies on pharmacologically active peptides and proteins from snake venoms have been concerned with isolation and structure elucidation through methods of classical biochemistry. As an attempt to examine the transcripts expressed in the venom gland of Bothrops jararacussu and to unveil the toxicological and pharmacological potential of its products at the molecular level, we generated 549 expressed sequence tags (ESTs) from a directional cDNA library. Sequences obtained from single-pass sequencing of randomly selected cDNA clones could be identified by similarities searches on existing databases, resulting in 197 sequences with significant similarity to phospholipase A(2) (PLA(2)), of which 83.2% were Lys49-PLA(2) homologs (BOJU-1), 0.1% were basic Asp49-PLA(2)s (BOJU-II) and 0.6% were acidic Asp49-PLA(2)s (BOJU-III). Adjoining this very abundant class of proteins we found 88 transcripts codifying for putative sequences of metalloproteases, which after clustering and assembling resulted in three full-length sequences: BOJUMET-I, BOJUMET-II and BOJUMET-III; as well as 25 transcripts related to C-type lectin like protein including a full-length cDNA of a putative galactose binding C-type lectin and a cluster of eight serine-proteases transcripts including a full-length cDNA of a putative serine protease. Among the full-length sequenced clones we identified a nerve growth factor (Bj-NGF) with 92% identity with a human NGF (NGHUBM) and an acidic phospholipase A2 (BthA-I-PLA(2)) displaying 85-93% identity with other snake venom toxins. Genetic distance among PLA(2)s from Bothrops species were evaluated by phylogenetic analysis. Furthermore, analysis of full-length putative Lys49-PLA(2) through molecular modeling showed conserved structural domains, allowing the characterization of those proteins as group II PLA(2)s. The constructed cDNA library provides molecular clones harboring sequences that can be used to probe directly the genetic material from gland venom of other snake species. Expression of complete cDNAs or their modified derivatives will be useful for elucidation of the structure-function relationships of these toxins and peptides of biotechnological interest. (C) 2004 Elsevier SAS. All rights reserved.

Phospholipase A(2) myotoxins from Bothrops snake venoms: Structure-function relationship

Phospholipases A(2) constitute the major components from Bothrops snake venoms and have been extensively investigated not only because they are relatively very abundant in these venoms but mainly because they display a range of many relevant biological effects, including: myotoxic, cytotoxic, edema-inducing, artificial membrane disrupting, anticoagulant, neuromuscular, platelet aggregation inhibiting, hypotensive, bactericidal, anti-HIV, anti-tumoural, anti-malarial and anti-parasitic. The primary structures of several PLA(2)s have been elucidated through direct amino acid sequencing or, inderectly, through the corresponding nucleotide sequencing. Two main subgroups were thus described: (i) Asp49 PLA(2)s, showing low (basic, highly myotoxic) to relatively high (acidic, less or non myotoxic) Ca++-dependent hydrolytic activity upon artificial substrates; (ii) Lys49 PLA(2)s (basic, highly myotoxic) , showing no detectable hydrolytic activity on artificial substrates. Several crystal structures of Lys49 PLAs from genus Bothrops have already been solved, revealing very similar fold patterns. Lack of catalytic activity of myotoxic Lys49-PLA(2)s, first related solely with the fact that Lys49 occupies the position of the calcium ion in the catalyticly active site of Asp49 PLA(2)s, is now also attributed to Lys122 which interacts with the carbonyl of Cys29 hyperpolarising the peptide bond between Cys29 and Gly30 and trapping the fatty acid product in the active site, thus interrupting the catalytic cycle. This hypothesis, supported for three recent structures, is also discussed here. All Asp49 myotoxins showed to be pharmacologically more potent when compared with the Lys49 variants, but phospholipid hydrolysis is not an indispensable condition for the myotoxic, cytotoxic, bactericidal, anti-HIV, anti-parasitic, liposome disrupting or edema-inducing activities. Recent studies on site directed mutagenesis of the recombinant Lys49 myotoxin from Bothrops jararacussu revealed the participation of important amino acid residues in the membrane damaging and myotoxic activities.

Rosmarinic acid, a new snake venom phospholipase A(2) inhibitor from Cordia verbenacea (Boraginaceae): antiserum action potentiation and molecular interaction

Many plants are used in traditional medicine as active agents against various effects induced by snakebite. The methanolic extract from Cordia verbenacea (Cv) significantly inhibited paw edema induced by Bothrops jararacussu snake venom and by its main basic phospholipase A(2) homologs, namely bothropstoxins I and II (BthTXs). The active component was isolated by chromatography on Sephadex LH-20 and by RP-HPLC on a C18 column and identified as rosmarinic acid (Cv-RA). Rosmarinic acid is an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid [2-O-cafeoil-3-(3,4-di-hydroxy-phenyl)-R-lactic acid]. This is the first report of RA in the species C. verbenacea ('baleeira', 'whaler') and of its anti-inflammatory and antimyotoxic properties against snake venoms and isolated toxins. RA inhibited the edema and myotoxic activity induced by the basic PLA(2)s BthTX-I and BthTX-II. It was, however, less efficient to inhibit the PLA(2) activity of BthTX-II and, still less, the PLA(2) and edema-inducing activities of the acidic isoform BthA-1-PLA(2), from the same venom, showing therefore a higher inhibitory activity upon basic PLA(2)s. RA also inhibited most of the myotoxic and partially the edema-inducing effects of both basic PLA(2)s, thus reinforcing the idea of dissociation between the catalytic and pharmacological domains. The pure compound potentiated the ability of the commercial equine polyvalent antivenom in neutralizing lethal and myotoxic effects of the crude venom and of isolated PLA(2)s in experimental models. CD data presented here suggest that, after binding, no significant conformation changes occur either in the Cv-RA or in the target PLA(2). A possible model for the interaction of rosmarinic acid with Lys49-PLA(2) BthTX-I is proposed. (c) 2005 Elsevier Ltd. All rights reserved.

Structure of BthA-I complexed with p-bromophenacyl bromide: possible correlations with lack of pharmacological activity

The crystal structure of an acidic phospholipase A(2) isolated from Bothrops jararacussu venom (BthA-I) chemically modified with p-bromophenacyl bromide (BPB) has been determined at 1.85 angstrom resolution. The catalytic, platelet-aggregation inhibition, anticoagulant and hypotensive activities of BthA-I are abolished by ligand binding. Electron-density maps permitted unambiguous identification of inhibitor covalently bound to His48 in the substrate-binding cleft. The BthA-I-BPB complex contains three structural regions that are modified after inhibitor binding: the Ca2+-binding loop, ss-wing and C-terminal regions. Comparison of BthA-I-BPB with two other BPB-inhibited PLA(2) structures suggests that in the absence of Na+ ions at the Ca2+- binding loop, this loop and other regions of the PLA(2)s undergo structural changes. The BthA-I-BPB structure reveals a novel oligomeric conformation. This conformation is more energetically and conformationally stable than the native structure and the abolition of pharmacological activities by the ligand may be related to the oligomeric structural changes. A residue of the `pancreatic' loop (Lys69), which is usually attributed as providing the anticoagulant effect, is in the dimeric interface of BthA-I-BPB, leading to a new hypothesis regarding the abolition of this activity by BPB.

Preliminary X-ray crystallographic studies of BthTX-II, a myotoxic Asp49-phospholipase A(2) with low catalytic activity from Bothrops jararacussu venom

For the first time, a complete X-ray diffraction data set has been collected from a myotoxic Asp49-phospholipase A(2) (Asp49-PLA(2)) with low catalytic activity (BthTX-II from Bothrops jararacussu venom) and a molecular-replacement solution has been obtained with a dimer in the asymmetric unit. The quaternary structure of BthTX-II resembles the myotoxin Asp49-PLA(2) PrTX-III (piratoxin III from B. pirajai venom) and all non-catalytic and myotoxic dimeric Lys49-PLA(2)s. In contrast, the oligomeric structure of BthTX-II is different from the highly catalytic and non-myotoxic BthA-I (acidic PLA(2) from B. jararacussu). Thus, comparison between these structures should add insight into the catalytic and myotoxic activities of bothropic PLA(2)s.