Giardia duodenalis: protein substrates degradation by trophozoite proteases

The present investigation was undertaken to identify and characterize trophozoite proteases of five axenic strains of Giardia duodenalis isolated in Brazil and the reference strain Portland 1 isolated in the United States. Trophozoite cell lysates of each strain were analysed for the pattern of proteins and for proteolytic activity. Samples were tested in SDS-polyacrylamide gel electrophoresis for the protein profiles, and the detection of proteases in cell lysates was performed using substrate gel electrophoresis [gelatin, collagen, bovine serum albumin (BSA) and haemoglobin] and azocasein assays. Indeed, synthetic inhibitors were included in the assays to characterize the protease classes. Differences on the hydrolysis patterns of protein substrates were observed in relation to the substrate composition as much as the Giardia trophozoite strain. The substrate-containing gels revealed hydrolysis bands with molecular masses ranging from > 97 to 20-15 kDa, and most zones were common to the five strains. However, some pronounced differences could be detected in the BTU-11 pattern. Azocasein was also degraded; however, depending on the lysate assayed, the degree of substrate degradation was variable. It was observed that inhibitory effects are substrate-dependent since the activity was predominantly due to cysteine proteases against gelatin, collagen, BSA and azocasein substrates and due to serine against haemoglobin. The presence of aspartic protease and aminopeptidase activity in the lysates was also indicated.

Colonization of rocky and leaf pack substrates by benthic macroinvertebrates in a stream in southeast Brazil.

A comparative analysis was conducted of the colonization by benthic macroinvertebrates of rocky and leaf pack substrates, both natural and artificial. This colonization was evaluated by season, with the objective of ascertaining the influence of rainfall on the rate of colonization. The total density of macroinvertebrates after 21 days of colonization was significantly greater in the dry than in the wet season. When the substrate types were compared, artificial leaf pack substrate presented the smallest density for both seasons. In the wet season, Chironomidae, Leptohyphidae, Hydropsychidae, Elmidae, immature stages of Trichoptera, and Hydroptilidae showed a more representative density. In the dry season, Chironomidae, Baetidae and Oligochaeta were the most abundant taxa. The artificial rocky substrate used in this experiment was the most appropriate, due to its resemblance with natural substrate conditions in terms of the maintenance of the structural integrity of the substrate throughout the experimental period. Successional and seasonal effects were of great relevance, playing an important role in the colonization process.

Tunnelling behaviour of the Asian subterranean termite in heterogeneous soils: presence of cues in the foraging area

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

Biomonitoring of substrates containing sewage sludge: assessment of the feasibility in using the diplopod Rhinocricus padbergi.

Due to the constant release of potentially harmful substances in the environment, it is necessary the use of biomonitoring as a form to assess the impact of these substances on animals that inhabit this environment. Therefore, there is a better understanding of the possible effects that these exogenous substances may cause in the organisms present there. In the present study it was verified the feasibility of the use of the diplopod Rhinocricus padbergi as bioindicator of impacted soils by their exposition to substrates containing different concentrations of sewage sludge from different Sewage Treatment Stations (STSs). It was observed animals' behavior and the survival rate was analyzed by log-rank test (p=0,05). The analysis showed that the animals exposed to pure sewage sludge presented higher mortality index than the specimens exposed to different concentrations of the sludge mixed with soil. In general, the survival rate and the analysis of the behavior of the diplopod R. padbergi showed efficiency of this species in biomonitoring impacted soils.

Influence of Different Substrates on the Production of a Mutant Thermostable Glucoamylase in Submerged Fermentation

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

Production of cyclodextrins by CGTase from Bacillus clausii using different starches as substrates

Cyclodextrins ( CDs) are cyclic oligasaccharides composed by D- glucose monomers joined by alpha- 1,4-D glicosidic linkages. The main types of CDs are alpha-,beta-and gamma-CDs consisting of cycles of six, seven, and eight glucose monomers, respectively. Their ability to form inclusion complexes is the most important characteristic, allowing their wide industrial application. The physical property of the CD-complexed compound can be altered to improve stability, volatility, solubility, or bio-availability. The cyclomaltodextrin glucanotransferase ( CGTase, EC 2.4.1.19) is an enzyme capable of converting starch into CD molecules. In this work, the CGTase produced by Bacillus clausii strain E16 was used to produce CD from maltodextrin and different starches ( commercial soluble starch, corn, cassava, sweet potato, and waxy corn starches) as substrates. It was observed that the substrate sources influence the kind of CD obtained and that this CGTase displays a beta- CGTase action, presenting a better conversion of soluble starch at 1.0%, of which 80% was converted in CDs. The ratio of total CD produced was 0: 0.89: 0.11 for alpha/beta/gamma. It was also observed that root and tuber starches were more accessible to CGTase action than seed starch under the studied conditions.

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.

Flexural Strength of Four Adhesive Fixed Dental Prostheses of Composite Resin Reinforced with Glass Fiber

Purpose: To evaluate the flexural strength of two fixed dental prosthesis (FDP) designs simulating frameworks of adhesive fixed partial prostheses, reinforced or not by glass fiber.Materials and Methods: Forty specimens, made with composite resin, were divided into 4 groups according to the framework design and the presence of fiber reinforcement: A1 - occlusal support; A2: occlusal support + glass fiber; B1: occlusal and proximal supports; B2: occlusal and proximal supports + glass fiber. The specimens were subjected to the three-point bending test, and the data were submitted to two-way ANOVA and Tukey's test (5%).Results: Group A2 (97.9 +/- 38 N) was statistically significantly different from all other experimental groups, presenting a significantly lower mean flexural strength.Conclusion: The use of glass fibers did not improve the flexural strength of composite resin, and designs with occlusal and proximal supports presented better results than designs simulating only occlusal support.

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.

Flexural Strength of Glass-Infiltrated Zirconia/Alumina-Based Ceramics and Feldspathic Veneering Porcelains

Purpose: To compare the flexural strength of two glass-infiltrated high-strength ceramics and two veneering glass-ceramics.Materials and Methods: Four ceramic materials were tested: two glass-infiltrated high-strength ceramics used as framework in metal-free restorations [In-Ceram Zirconia IZ (Gr1) and In-Ceram Alumina IA (Gr2)], and two glass-ceramics used as veneering material in metal-free restorations [Vita VM7 (Gr3) and Vitadur-alpha (Gr4)]. Bar specimens (25 x 5 x 2 mm(3)) made from core ceramics, alumina, and zirconia/alumina composites were prepared and applied to a silicone mold, which rested on a base from a gypsum die material. The IZ and IA specimens were partially sintered in an In-Ceram furnace according to the firing cycle of each material, and then were infiltrated with a low-viscosity glass to yield bar specimens of high density and strength. The Vita VM7 and Vitadur-alpha specimens were made from veneering materials, by vibration of slurry porcelain powder and condensation into a two-part brass Teflon matrix (25 x 5 x 2 mm(3)). Excess water was removed with absorbent paper. The veneering ceramic specimens were then removed from the matrix and were fired as recommended by the manufacturer. Another ceramic application and sintering were performed to compensate the contraction of the feldspar ceramic. The bar specimens were then tested in a three-point bending test.Results: The core materials (Gr1: 436.1 +/- 54.8; Gr2: 419.4 +/- 83.8) presented significantly higher flexural strength (MPa) than the veneer ceramics (Gr3: 63.5 +/- 9.9; Gr4: 57.8 +/- 12.7).Conclusion: In-Ceram Alumina and Zirconia were similar statistically and more resistant than VM7 and Vitadur-alpha.

Effect of Acid Neutralization and Mechanical Cycling on the Microtensile Bond Strength of Glass-ceramic Inlays

Objectives: To evaluate the hypothesis that a process of hydrofluoric acid precipitate neutralization and fatigue load cycling performed on human premolars restored with ceramic inlays had an influence on microtensile bond strength results (MTBS). Methods: MOD inlay preparations were performed in 40 premolars (with their roots embedded in acrylic resin). Forty ceramic restorations were prepared using glass-ceramic (IPS Empress). The inner surfaces of all the restorations were etched with 10% hydrofluoric acid for 60 seconds, rinsed with water and dried. The specimens were divided into two groups (N=20): 1-without neutralization; 2-with neutralization. All the restorations were silanized and adhesively cemented (self-curing and self-etching luting composite system, Multilink). Ten premolars from each group were submitted to mechanical cycling (1,400,000 cycles, 50N, 37 degrees C). After cycling, the samples were sectioned to produce non-trimmed beam specimens (vestibular dentin-restoration-lingual dentin set), which were submitted to microtensile testing. Results: Bond strength was significantly affected by the surface treatment (p<0.0001) (no neutralization > neutralization) and mechanical cycling (p<0.0001) (control > cycling) (2-way ANOVA and Tukey test, alpha=.05). Conclusion: Hydrofluoric acid precipitate neutralization appears to significantly damage the resin bond to glass-ceramic and should not be recommended. The clinical simulation of the specimens, by using mechanical cycling, is important when evaluating the ceramic-dentin bond.

Effect of Carbamide Peroxide Bleaching Gel Concentration on the Bond Strength of Dental Substrates and Resin Composite

Purpose: This study evaluated the effect of bleaching gel containing 10%, 15% and 20% carbamide peroxide (CP) on the bond strength of dental enamel or dentin and resin composite restorations.Methods: The buccal surfaces of 12 bovine tooth crowns were conditioned with 37% phosphoric acid, and the adhesive resin Single Bond 2 and the resin composite Filtek Z350 were used to perform the restorations. The blocks were sectioned to obtain bar specimens. Each specimen group (enamel-E, dentin-D) was divided into four subgroups (n=15): S-artificial saliva; 10-10% CP bleaching; 15-15% CP bleaching; 20-20% CP bleaching. CP was applied for six hours daily for two weeks. The specimens were submitted to the a test in a universal testing machine. The data were analyzed by one-way ANOVA and the Tukey post-hoc test and a correlation analysis (r) was performed.Results: For Group E, the mean value (+/- standard-deviation) was 21.86 (+/- 6.03)a, 18.91 (+/- 8.31)ab, 15.43 (+/- 7.44)b and 10.6 (+/- 4.94)c for ES, E10, E15 and E20, respectively. For Group D, the alpha values were 34.73 (+/- 4.68)a, 35.12 (+/- 13.43)a, 29.67 (+/- 6.84)ab and 24.56 (+/- 6.54)b for DS, D10, D15 and D20, respectively. A negative correlation between the CP concentration and mean values was observed for both the enamel (r=-0.95) and dentin (r=-0.85) groups.Conclusion: In the current study, the bond strength of the restoration to enamel and the restoration to dentin were influenced by the application of CP and was dependent on the CP concentration in the bleaching gel.

Surface degradation of glass ceramics after exposure to acidulated phosphate fluoride

Objective: This study evaluated the surface degradation effect of acidulated phosphate fluoride (APF) gel exposure on the glassy matrix ceramics as a function of time. Material and methods: Disc-shaped ceramic specimens (N = 120, 10/per ceramic material) were prepared in stainless steel molds (inner diameter: 5 mm, height: 2 mm) using 6 dental ceramics: 3 indicated for ceramic-fused-to-metal (Vita Omega 900, Carmen and Vita Titankeramik), 2 for all-ceramic (Vitadur Alpha and Finesse (R) Low Fusing) and 1 for both types of restorations (IPS d. SIGN). The specimens were wet ground finished, ultrasonically cleaned and auto-glazed. All specimens were subjected to calculation of percentage of mass loss, surface roughness analysis and topographical description by scanning electron microscopy (SEM) before (0 min) and after exposure to 1.23 % APF gel for 4 min and 60 min representing short-and long-term etching effect, respectively. The data were analyzed using two-way ANOVA with repeated measures and Tukey` s test (alpha=0.05). Results: Significant effect of the type of the ceramics (p=0.0000, p=0.0031) and exposure time (p=0.0000) was observed in both surface roughness and percentage of mass loss values, respectively. The interaction factor between both parameters was also significant for both parameters (p=0.0904, p=0.0258). Both 4 min (0.44 +/- 0.1-0.81 +/- 0.2 mu m) and 60 min (0.66 +/- 0.1 - 1.04 +/- 0.3 mu m) APF gel exposure created significantly more surface roughness for all groups when compared to the control groups (0.33 +/- 0.2-0.68 +/- 0.2 mu m) (p<0.05). There were no significant differences in percentage of mass loss between the ceramics at 4 min (p>0.05) but at 60 min exposure, IPS d. SIGN showed the highest percentage of mass loss (0.1151 +/- 0.11). The mean surface roughness for Vita Titankeramik (0.84 +/- 0.2 mu m) and Finesse (R) Low Fusing (0.74.+/- 0.2 mu m) was significantly higher than those of the other ceramics (0.59 +/- 0.1 mu m - 0.49 +/- 0.1 mu m) and Vita Titankeramik (p<0.05) regardless of the exposure time. A positive correlation was found between surface roughness and percentage of mass loss for all ceramic materials [(r=0.518 (Vitadur Alpha), r=0.405 (Vita Omega 900), r=0.580 (Carmen), r=0.687 (IPS d. SIGN), r=0.442 (Finesse (R) Low Fusing), r=0.572 (Vita Titankeramik), Pearson's correlation coefficient)]. The qualitative SEM analysis showed evidence of corrosive attack on all of ceramics at varying degrees. Conclusions: The ceramics indicated for either metal-ceramic or all-ceramic restorations were all vulnerable to surface texture changes and mass loss after short-term and long-term APF gel exposure.