Ultrastructure of Giardia duodenalis trophozoites group from hamster: some relevant aspects

Throphozoites of Giardia duodenalis group obtained from fragments or scratched of hamster's mucosa were examined by transission electron microscopy. The fine structure of the trophozoites are presented and comapred with those described for other animals. Some of the trophozoites present the cytoplasm full of glycogen, rough endoplasmic reticulum-like structures and homogeneous inclusions not enclosed by membranes, recognized as lipid drops, which had not been observed in Giardia from other animals. The adhesive disk is composed of a layer of microtubules, from which fibrous ribbons extend into the cytoplasm; these ribbons are linked by layer of crossbridge filaments that shows an intermediary dense band, described for the first time in this paper. The authors regard this band as the result of the cross-bridge filaments slinding in the medium region between adjacent fibrous ribbons, and suggest a contractile activity for them. The role of the adhesive disk on the trophozoite mechanism of attachment to host mucosa is also discussed.

Structure determination of a tetrahydro-beta-carboline of arthropod origin: A novel alkaloid-toxin subclass from the web of spider Nephila clavipes

The orb-web spiders are polyphagous animals in which the web plays a very important role in the capture of preys; oily droplets usually cover the capture-web of the spider Nephila clavipes and seem to be of great importance for prey capture. The knowledge of the chemical composition of these droplets is necessary to understand the function of this adhesive material in web mechanics and prey capture. A novel subclass of spider toxins, tetrahydro-beta-carboline, was identified among the weaponry of compounds present inside of oily droplets. This type of alkaloid is not common among the natural compounds of spider toxins. Apparently, when the prey arthropods get caught by the spider web, their bodies are covered with many adhesive oily droplets, which disrupt delivering the tetrahydro-beta-carboline to the direct contact with the prey integument. Toxicity assays demonstrated a potent lethal effect of the alkaloid toxin to the spider preys; topical applications of the teirahydro-beta-carboline at first caused clear signs of neurotoxicity, followed by the death of preys. The structure of the major component, a tetrahydro-beta-carboline, among the alkaloid toxins was elucidated by means of UV spectrophotometry, ESI mass spectrometry, H-1-NMR spectroscopy, and high-resolution mass spectrometry. The structure of the natural toxin was determined as 1-(2-guanidinoethyl)-1,2,3,4-tetrahydro-6-hydroxymethyl)-beta-carboline; the investigation of the pharmacological properties and neurotoxic actions of this compound may be used in the future as reference for the development of new drugs to be applied at level of pest control in agriculture.

The chelation of metal ions by the acylpolyamine toxins from the web-spider Nephilengys cruentata: effects in the intoxication/detoxification of preys

Orb-web-spiders present a series of different strategies for prey capture, involving the use of different types of silk for web building, the use of adhesive traps in the webs, the secretion of toxic compounds to the spider's preys in the adhesive coating of the capture web and the biosynthesis of a wide range of structurally related acylpolyamine toxins in their venoms. The polyamine toxins usually block neuromuscular junctions and/or the central nervous system (CNS) of Arthropods, targeting specially the ionotropic glutamate receptors; this way these toxins are used are as chemical weapons to kill / paralyze the spider's prey. Polyamine toxins contain many azamethylene groups involved with the chelation of metal ions, which in turn can interact with the glutamate receptors, affecting the toxicity of these toxins. It was demonstrated that the chelation of Ni+2, Fe+2, Pb+2, Ca+2 and Mg+2 ions by the desalted crude venom of Nephilengys cruentata and by the synthetic toxin JSTX-3, did not cause any significant change in the toxicity of the acylpolyamine toxins to the model-prey insect (honeybees). However, it was also reported that the chelation of Zn+2 ions by the acylpolyamines potentiated the lethal / paralytic action of these toxins to the honeybees, while the chelation of Cu+2 ions caused the inverse effect. Atomic absorption spectrometry and Plasma-ICP analysis both of N.cruentata venom and honeybee's hemolymph revealed that the spider's venom concentrates Zn+2 ions, while the honeybee's hemolymph concentrates Cu+2 ions. These results are suggesting that the natural accumulation of Zn+2 ions in N. cruentata venom favors the prey catching and/or its maintenance in the web, while the natural accumulation of Cu+2 ions in prey's hemolymph minimizes the efficiency of the acylpolyamine toxins as killing/paralyzing tool.

A rapid technique for analysis of formalin-fixed, paraffin-embedded tissues by fluorescent in situ hybridization with alpha-satellite probes

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

Effect of Airborne-Particle Abrasion and Mechanico-Thermal Cycling on the Flexural Strength of Glass Ceramic Fused to Gold or Cobalt-Chromium Alloy

Purpose: To evaluate the effect of airborne-particle abrasion and mechanico-thermal cycling on the flexural strength of a ceramic fused to cobalt-chromium alloy or gold alloy.Materials and Methods: Metallic bars (n = 120) were made (25 mm x 3 mm x 0.5 mm): 60 with gold alloy and 60 with Co-Cr. At the central area of the bars (8 mm x 3 mm), a layer of opaque ceramic and then two layers of glass ceramic (Vita VM13, Vita Zahnfabrick) were fired onto it (thickness: 1 mm). Ten specimens from each alloy group were randomly allocated to a surface treatment [(tungsten bur or air-particle abrasion (APA) with Al(2)O(3) at 10 mm or 20 mm away)] and mechanico-thermal cycling (no cycling or mechanically loaded 20,000 cycles; 10 N distilled water at 37 degrees C and then thermocycled 3000 cycles; 5 degrees C to 55 degrees C, dwell time 30 seconds) combination. Those specimens that did not undergo mechanico-thermal cyclingwere stored inwater (37 degrees C) for 24 hours. Bond strength was measured using a three-point bend test, according to ISO 9693. After the flexural strength test, failure types were noted. The data were analyzed using three factor-ANOVA and Tukey's test (alpha = 0.05).Results: There were no significant differences between the flexural bond strength of gold and Co-Cr groups (42.64 +/- 8.25 and 43.39 +/- 10.89 MPa, respectively). APA 10 and 20 mm away surface treatment (45.86 +/- 9.31 and 46.38 +/- 8.89 MPa, respectively) had similar mean flexural strength values, and both had significantly higher bond strength than tungsten bur treatment (36.81 +/- 7.60 MPa). Mechanico-thermal cycling decreased the mean flexural strength values significantly for all six alloy-surface treatment combinations tested when compared to the control groups. The failure type was adhesive in the metal/ceramic interface for specimens surface treated only with the tungsten bur, and mixed for specimens surface treated with APA 10 and 20 mm.Conclusions: Considering the levels adopted in this study, the alloy did not affect the bond strength; APA with Al(2)O(3) at 10 and 20 mm improved the flexural bond strength between ceramics and alloys used, and the mechanico-thermal cycling of metal-ceramic specimens resulted in a decrease of bond strength.

SEALING ABILITY of CASTOR OIL POLYMER AS A ROOT-END FILLING MATERIAL

Objective: The purpose of this study was to evaluate the sealing ability of castor oil polymer (COP), mineral trioxide aggregate (MTA) and glass ionomer cement (GIC) as root-end filling materials. Forty-five single-rooted human teeth were cleaned and prepared using a step-back technique. The apical third of each root was resected perpendicularly to the long axis direction. All teeth were obturated with gutta-percha and an endodontic sealer. After, a root-end cavity with 1.25-mm depth was prepared using a diamond bur. The specimens were randomly divided into three experimental groups (n = 15), according to the root-end filling material used: G1) COP; G2) MTA; G3) GIC. The external surfaces of the specimens were covered with epoxy adhesive, except the root-end filling. The teeth were immersed in rhodamine B dye for 24 hours. Then, the roots were sectioned longitudinally and the linear dye penetration at the dentin/material interface was determined using a stereomicroscope. ANOVA and Tukey's tests were used to compare the three groups. The G1 group (COP) presented smaller dye penetration, statistically different than the G2 (MTA) and G3 (GIC) groups (p < 0.05). No statistically significant difference in microleakage was observed between G2 and G3 groups (p > 0.05). The results of this study indicate that the COP presented efficient sealing ability when used as a root-end filling material showing results significantly better than MTA and GIC.

Homogenous amniotic membrane as a biological dressing for oral mucositis in rats: Histomorphometric analysis

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

Avaliação qualitativa do efeito de agentes de limpeza na camada de lama dentinária: estudo ultra-estrutural em microscopia eletrônica de varredura

Quando qualquer instrumento abrasiona ou corta a dentina, produz na superfície uma camada de lama dentinária ou smear layer. Dependendo do agente de união indicado em Odontologia adesiva, há a necessidade ou não da remoção da camada de lama da superfície dentinária. Com a finalidade de verificar a ação de diferentes substâncias para a limpeza dentinária, utilizamos 20 dentes pré-molares superiores íntegros, mantidos em soro fisiológico, nos quais as coroas foram seccionadas ao meio no sentido mésio-distal. Com instrumento diamantado, removeu-se o esmalte da porção vestibular e da porção lingual da coroa e, com uma broca carbide cilíndrica lisa nº 56, cortou-se aproximadamente 1 mm de dentina com alta rotação sob abundante refrigeração ar/água, para produzir a camada de lama dentinária. em seguida, essa superfície foi tratada com diferentes substâncias e lavada por 30 segundos com spray ar/água. No controle, foi simplesmente utilizado o spray ar/água. Os espécimes foram montados em suportes metálicos, preparados e visualizados no MEV-DSM 950 da Zeiss, em aumentos que variaram de 100 a 5.000 vezes. Os materiais que mais removeram a camada de lama foram, em ordem crescente: 1. spray ar/água; 2. fluoreto de sódio 2%; 3. associação alternada de Dakin/Tergensol; 4. água oxigenada 3%; 5. jateamento com óxido de alumínio 50 mm; 6. flúor acidulado 1,27%; 7. ácido poliacrílico 25%; 8. ácido fosfórico 10%.

Effect of conditioning methods on the microtensile bond strength of phosphate monomer-based cement on zirconia ceramic in dry and aged conditions

The objective of this study was to evaluate the durability of bond strength between a resin cement and aluminous ceramic submitted to various surface conditioning methods. Twenty-four blocks (5 X 5 X 4 mm 3) of a glass-in filtrated zirconia-alumina ceramic (inCeram Zirconia Classic) were randomly divided into three surface treatment groups: ST1-Air-abrasion with 110-mu m Al2O3 particles + silanization; ST2-Laboratory tribochemical silica coating method (110-mu m Al2O3, ilO-PM Silica) (Rocatec) + silanization; ST3-Chairside tribochemical silica coating method (30-mu m SiOx) (CoJet) + silanization. Each treated ceramic block was placed in its silicone mold with the treated surface exposed. The resin cement (Panavia F) was prepared and injected into the mold over the treated surface. Specimens were sectioned to achieve nontrimmed bar specimens (14 sp/block) that were randomly divided into two conditions: (a) Dry-microtensile test after sectioning; (b) Thermocycling (TC)-(6,000X, 5-55 degrees C) and water storage (150 days). Thus, six experimental groups were obtained (11 = 50): Gr1-ST1 + dry; Gr2-ST1 + TC. Gr3-ST2 + dry; Gr4-ST2 + TC; Gr5-ST3 + dry; Gr6ST3 + TC. After microtensile testing, the failure types were noted. ST2 (25.1 +/- 11) and ST3 (24.1 +/- 7.4) presented statistically higher bond strength (MPa) than that of STI (17.5 +/- 8) regardless of aging conditions (p < 0.0001). While Gr2 revealed the lowest results (13.3 +/- 6.4), the other groups (21.7 +/- 7.4-25. 9 +/- 9.1) showed statistically no significant differences (two-way ANOVA and Tukey's test, a 0.05). The majority of the failures were mixed (82%) followed by adhesive failures (18%). Gr2 presented significantly higher incidence of ADHESIVE failures (54%) than those of other groups (p = 0.0001). Both laboratory and chairside silica coating plus silanization showed durable bond strength. After aging, airabrasion with 110-mu m Al2O3 + silanization showed the largest decrease indicating that aging is fundamental for bond strength testing for acid-resistant Arconia ceramics in order to estimate their long-term performance in the mouth. (c) 2007 Wiley Periodicals, Inc.

The Influence of Cutting Speed and Cutting Initiation Location in Specimen Preparation for the Microtensile Bond Strength Test

Purpose: This study evaluated the effect of cutting initiation location and cutting speed on the bond strength between resin cement and feldspathic ceramic.Materials and Methods: Thirty-six blocks (6.4 x 6.4 x 4.8 mm) of ceramic (Vita VM7) were produced. The ceramic surfaces were etched with 10% hydrofluoric acid gel for 60 s and then silanized. Each ceramic block was placed in a silicon mold with the treated surface exposed. A resin cement (Variolink II) was injected into the mold over the treated surface and polymerized. The resin cement-ceramic blocks were divided into two groups according to experimental conditions: a) cutting initiation location - resin cement, ceramic and interface; and b) cutting speed - 10,000, 15,000, and 20,000 rpm. The blocks were sectioned to achieve non-trimmed bar specimens. The microtensile test was performed in a universal testing machine (1 mm/min). The failure modes were examined using an optical light microscope and SEM. Bond strength results were analyzed using one-way ANOVA and Tukey's test (alpha = 0.05).Results: Significant influences of cutting speed and initiation location on bond strength (p < 0.05) were observed. The highest mean was achieved for specimens cut at 15,000 rpm at the interface (15.12 +/- 5.36 MPa). The lowest means were obtained for specimens cut at the highest cutting speed in resin cement (8.50 +/- 3.27 MPa), and cut at the lowest cutting speed in ceramic (8.60 +/- 2.65MPa). All groups showed mainly mixed failure (75% to 100%).Conclusion: The cutting speed and initiation location are important factors that should be considered during specimen preparation for microtensile bond strength testing, as both may influence the bond strength results.

Evaluation of interface characterization and adhesion of glass ceramics to commercially pure titanium and gold alloy after thermal- and mechanical-loading

Objectives. This study evaluated the effect of thermal- and mechanical-cycling on the shear bond strength of three low-fusing glassy matrix dental ceramics to commercial pure titanium (cpTi) when compared to conventional feldspathic ceramic fused to gold alloy.Methods. Metallic frameworks (diameter: 5 min, thickness: 4 mm) (N = 96, n = 12 per group) were cast in cpTi and gold alloy, airborne particle abraded with 150 mu m aluminum oxide. Low-fusing glassy matrix ceramics and a conventional feldspathic ceramic were fired onto the alloys (thickness: 4mm). Four experimental groups were formed; Gr1 (control group): Vita Omega 900-Au-Pd alloy; Gr2: Ticeram-cpTi; Gr3: Super Porcelain Ti-22-cpTi and G4: Vita Titankeramik-cpTi. While half of the specimens from each ceramic-metal combination were randomly tested without aging (water storage at 37 C for 24h only), the other half were first thermocycled (6000 cycles, between 5 and 55 C, dwell time: 13 s) and then mechanically loaded (20,000 cycles under SON load, immersion in distilled water at 37 C). The ceramic-alloy interfaces were loaded under shear in a universal test machine (cross-head speed: 0.5 mm/min) until failure occur-red. Failure types were noted and the interfaces of the representative fractured specimens from each group were examined with stereo microscope and scanning electron microscope (SEM). in an additional study (N = 16, n = 2 per group), energy dispersive X-ray spectroscopy (EDS) analysis was performed from ceramic-alloy interfaces. Data were analyzed using ANOVA and Tukey's test.Results. Both ceramic-metal combinations (p < 0.001) and aging conditions (p < 0,001) significantly affected the mean bond strength values. Thermal- and mechanical-cycling decreased the bond strength (MPa) results significantly for Gr3 (33.4 +/- 4.2) and Gr4 (32.1 +/- 4.8) when compared to the non-aged groups (42.9 +/- 8.9, 42.4 +/- 5.2, respectively). Gr1 was not affected significantly from aging conditions (61.3 +/- 8.4 for control, 60.7 +/- 13.7 after aging) (p > 0.05). Stereomicroscope images showed exclusively adhesive failure types at the opaque ceramic-cpTi interfacial zone with no presence of ceramic on the substrate surface but with a visible dark titanium oxide layer in Groups 2-4 except Gr1 where remnants of bonder ceramic was visible. EDS analysis from the interfacial zone for cpTi-ceramic groups showed predominantly 34.5-85.1% O(2) followed by 1.1-36.7% Aland 0-36.3% Si except for Super Porcelain Ti-22 where a small quantity of Ba (1.4-8.3%), S (0.7%) and Sn (35.3%) was found. In the Au-Pd alloy-ceramic interface, 56.4-69.9% O(2) followed by 15.6-26.2% Si, 3.9-10.9% K, 2.8-6% Na, 4.4-9.6% Al and 0-0.04% Mg was observed.Significance. After thermal-cycling for 6000 times and mechanical-cycling for 20,000 times, Triceram-cpTi combination presented the least decrease among other ceramic-alloy combinations when compared to the mean bond strength results with Au-Pd alloy-Vita Omega 900 combination. (c) 2008 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Microshear Bond Strength of Self-etching Bonding Systems to Ultrasound Diamond Bur-prepared Dentin

Purpose: The aim of this study was to evaluate the influence of dentin abrasion on the microshear bond strength of two self-etching adhesive systems, using either an ultrasound diamond bur or a high-speed diamond bur.Materials and Methods: Twenty noncarious human third molars were sectioned mesiodistally into halves. The enamel was ground to expose a flat dentin surface on both sections. The dentinal surfaces were randomly assigned to two groups, depending on the method of smear layer preparation: ultrasound diamond bur (UB) or conventional diamond bur (CB). The prepared dentin surfaces received one of two self-etching systems: Clearfil SE Bond (CF) and One-Up Bond F (OB). A composite cylinder with a 0.95-mm diameter was bonded to each specimen and the microshear bond test was performed. The results were expressed in MPa and were subjected to two-way analysis of variance (ANOVA) and Tukey's test (alpha = 0.05).Results: There was no significant difference in dentin bond strength when comparing the conventional and ultrasonic abrasion methods. When the adhesive systems were compared, Clearfil SE Bond achieved higher bond strength means than did One-Up Bond F.Conclusion: The dentin surface preparation method did not influence the microshear bond strength and the Clearfil SE Bond adhesive system, independent of bur type used, Clearfil SE Bond showed higher bond strengths than did the One-Up Bond F adhesive system.

Evaluation of light transmission through translucent and opaque posts

Objectives: The transmission of light through translucent posts was observed, and the microhardness of light-cured cement used to secure these posts was evaluated at different depths. Methods: Fifteen single-rooted standard bovine teeth, 16 mm in size, were used. The root canals were prepared using #3 drills Light-Post (five teeth) and Aestheti Post (five teeth) systems (BISCO), with a working-length of 12 mm. In five teeth, translucent posts were cemented (Light-Post #2), while another five teeth received opaque posts (Aestheti Post #2). The roots were painted with black nail varnish to prevent the passage of light through the lateral walls of the roots. The root canals of all the specimens were treated with the All-Bond 2 adhesive system (BISCO) and cemented with light-cured cement (Enforce, Dentsply). All the roots were transversally cut to obtain six specimens 1.5 mm thick. Every two sections corresponded to a specific region of the root (cervical, middle, apical), making it possible to observe the cement microhardness at different levels. The groups (n=10) were defined as: G1: translucent post (TP)/cervical region; G2: TP/middle region; G3: TP/apical region; G4: Opaque post (OP)/cervical region; G5: OP/middle region; G6: PO/apical region. Five root canals were only filled with cement for use as a control (G7). Then, Vickers microhardness analyses were performed. Results: In G3, G5 and G6, the cement was not sufficiently hard to allow for microhardness analysis. When submitted to the ANOVA test, G1 (35.07), G2 (24.28) and G4 (28.64) presented no statistical differences. When the previous groups were compared to G7 (51.00) using the Kruskal-Wallis test, a statistical difference was found. Conclusion: Translucent posts allow cement polymerization up to the middle portion of the root.

Resin Bonding to a Feldspar Ceramic After Different Ceramic Surface Conditioning Methods: Evaluation of Contact Angle, Surface pH, and Microtensile Bond Strength Durability

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

Bond strength durability of a resin composite on a reinforced ceramic using various repair systems

Objectives. This study compared the durability of repair bond strength of a resin composite to a reinforced ceramic after three repair systems.Methods. Alumina-reinforced feldspathic ceramic blocks (Vitadur-alpha(R)) (N=30) were randomly divided into three groups according to the repair method: PR-Porcelain Repair Kit (Bisco) [etching with 9.5% hydrofluoric acid + silanization + adhesive]; CJ-CoJet Repair Kit (3M ESPE) [(chairside silica coating with 30 mu m SiO2 + silanization (ESPE(R)-Sil) + adhesive (Visio(TM)-Bond)]; CL-Clearfil Repair Kit [diamond surface roughening, etching with 40% H3PO4 + Clearfil Porcelain Bond Activator + Clearfil SE Bond)]. Resin composite was photo-polymerized on each conditioned ceramic block. Non-trimmed beam specimens were produced for the microtensile bond strength (mu TBS) tests. In order to study the hydrolytic durability of the repair methods, the beam specimens obtained from each block were randomly assigned to two conditions. Half of the specimens were tested either immediately after beam production (Dry) or after long-term water storage (37 degrees C, 150 days) followed by thermocyling (12,000 cycles, 5-55 degrees C) in a universal testing machine (1 mm/min). Failure types were analyzed under an optical microscope and SEM.Results. mu TBS results were significantly affected by the repair method (p=0.0001) and the aging conditions (p=0.0001) (two-way ANOVA, Tukey's test). In dry testing conditions, PR method showed significantly higher (p < 0.001) repair bond strength (19.8 +/- 3.8 MPa) than those of CJ and CL (12.4 +/- 4.7 and 9.9 +/- 2.9, respectively). After long-term water storage and thermocycling, CJ revealed significantly higher results (14.5 +/- 3.1 MPa) than those of PR (12.1 +/- 2.6 MPa) (p < 0.01) and CL (4.2 +/- 2.1 MPa) (p < 0.001). In all groups when tested in dry conditions, cohesive failure in the composite accompanied with adhesive failure at the interface (mixed failures), was frequently observed (76%, 80%, 65% for PR, CJ and CL, respectively). After aging conditions, while the specimens treated with PR and CJ presented primarily mixed failure types (52% and 87%, respectively), CL group presented mainly complete adhesive failures at the interface (70%).Significance. Hydrolytic stability of the repair method based on silica coating and silanization was superior to the other repair strategies for the ceramic tested. (C) 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.