Branched chain amino acids catabolism as a source of new drug targets in pathogenic protists.

Leucine, isoleucine, and valine, collectively termed Branched Chain Amino Acids (BCAA), are hydrophobic amino acids (AAs) and are essential for most eukaryotes since in these organisms they cannot be biosynthesized and must be supplied by the diet. These AAs are structurally relevant for muscle cells and, of course, important for the protein synthesis process. The metabolism of BCAA and its participation in different biological processes in mammals have been relatively well described. However, for other organisms as pathogenic parasites, the literature is really scarce. Here we review the BCAA catabolism, compile evidence on their relevance for pathogenic eukaryotes with special emphasis on kinetoplastids and highlight unique aspects of this underrated pathway.

Effects of dietary fibre intake in chemotherapy-induced mucositis in murine model.

Mucositis is an inflammation of the gastrointestinal mucosa resulting from high doses of radio/chemotherapy treatment and may lead to interruption of antineoplasic therapy. Soluble fibres, like pectin, increase SCFA production, which play a role in gut homoeostasis and inflammation suppression. Due to the properties of pectin, the aim of the present study was to evaluate the effect of a high-fibre (HF) diet on chemotherapy-induced mucositis in a murine model. C57/BL6 mice received control (AIN93M), HF, low/zero fibre (LF) diets for 10 d prior to mucositis challenging with irinotecan (75 mg/kg), or they were treated with acetate added to drinking water 5 d prior to and during the mucositis induction. Mice that received the HF diet showed decreased immune cells influx and improved histopathological parameters in the intestine, compared with mice that received the normal diet. Furthermore, the HF diet decreased intestinal permeability induced in the mucositis model when compared with the control group. This effect was not observed for acetate alone, which did not improve gut permeability. For instance, mice that received the LF diet had worsened gut permeability, compared with mice that received the normal diet and mucositis. The effects of the HF and LF diets were shown to modulate the intestinal microbiota, in which the LF diet increased the levels of Enterobacteriaceae, a group associated with gut inflammation, whereas the HF diet decreased this group and increased Lactobacillus and Bifidobacterium (SCFA producers) levels. In conclusion, the results demonstrated the importance of dietary fibre intake in the modulation of gut microbiota composition and homoeostasis maintenance during mucositis in this model.

Enantioselective biotransformation of sterically hindered amine substrates by the fungus Stemphylium lycopersici.

AIMS: To screen 20 micro-organisms for ω-transaminase (ω-TA) activity by the kinetic resolution of rac-1-phenylethylamine, followed by testing rac-amines of pharmaceutical interest with bulky substituents and to conduct the asymmetric synthesis of a chiral amine. METHODS AND RESULTS: Stemphylium lycopersici was selected as the best biocatalyst. By the central composite rotatable design (CCRD), it was found that, at lower pH (5·5 and 6·5), the lyophilized micro-organism biocatalysed the kinetic resolution of rac-1-phenylethylamine with 99% enantiomeric excess (e.e.) ((R)-enantiomer) with acetophenone conversions ranged from 41 to 45%. Interestingly, the lyophilized crude enzymatic extract lead to better results at pH from 7·0 to 9·0, with conversions up to 47% and about 99% e.e. We also attested that as much as higher is the pyruvate (amino acceptor) concentration, higher is the acetophenone conversion, corroborating the presence of ω-TA-type enzymes. Among different sterically hindered racemic amines tested, rac-1,2,3,4-tetrahydro-1-naphthylamine and rac-phenylbutylamine were satisfactorily kinetically resolved in up to 91% e.e. (R). The results for the asymmetric synthesis showed excellent conversion (>85%) for the S-1-phenylethylamine, indicating (S)-stereopreference. CONCLUSION: Stemphylium lycopersici showed to be an important tool for broader substrate scope transaminases and a relevant player on the development of new biocatalysts with ability in asymmetric synthesis reactions. SIGNIFICANCE AND IMPACT OF THE STUDY: Here in, we contribute to the improvement of the biocatalytic toolbox for chiral amines synthesis. Interestingly, we have found that the crude enzymatic extract of the endophytic fungus S. lycopersici could accept bulky substrates with reasonable activity, compared to the wild-type transaminase already published over literature, and with high enantioselectivity.

Modeling of coastal water contamination in Fortaleza (Northeastern Brazil).

An important tool in environmental management projects and studies due to the complexity of environmental systems, environmental modeling makes it possible to integrate many variables and processes, thereby providing a dynamic view of systems. In this study the bacteriological quality of the coastal waters of Fortaleza (a state capital in Northeastern Brazil) was modeled considering multiple contamination sources. Using the software SisBaHiA, the dispersion of thermotolerant coliforms and Escherichia coli from three sources of contamination (local rivers, storm drains and submarine outfall) was analyzed. The models took into account variations in bacterial decay due to solar radiation and other environmental factors. Fecal pollution discharged from rivers and storm drains is transported westward by coastal currents, contaminating strips of beach water to the left of each storm drain or river. Exception to this condition only occurs on beaches protected by the breakwater of the harbor, where counterclockwise vortexes reverse this behavior. The results of the models were consistent with field measurements taken during the dry and the rainy season. Our results show that the submarine outfall plume was over 2 km from the nearest beach. The storm drains and the Maceió stream are the main factors responsible for the poor water quality on the waterfront of Fortaleza. The depollution of these sources would generate considerable social, health and economic gains for the region.

Resin bonding to root dentin: influence of the alveolar bone level and thickness of the cement layer.

AIM: Aim of the present study was to evaluate the influence of the alveolar bone level and thickness of the cement layer on the push-out bond strength of fiber posts luted to root dentin. METHODS: We prepared post spaces in 60 bovine roots (16 mm), embedded in acrylic resin to simulate different scenarios of alveolar bone resorption. They were divided into six groups (N.=10), according to the factors "alveolar bone level" (14, 10, and 7 mm) and "cement thickness" (post no. 3/control and post no. 1/ thicker cement layer): G14P3 - 14-mm root + post n. 3; G10P3 - 10-mm root + post n. 3; G7P3 - 7-mm root + post n. 3; G14P1 - 14-mm root + post n. 1; G10P1 - 10-mm root + post n. 1; and G7P1 - 7-mm root + post n. 1. The fiber posts (White Post DC, FGM) were luted, and composite cores were made. Each specimen was subjected to mechanical cycling, and four slices were obtained from each root specimen and subjected to push-out testing in a universal testing machine. The data (MPa) were analyzed by ANOVA (two way) and Tukey's test (5%). RESULTS: The factor "cement thickness" (P=0.0057) significantly influenced bond strength, but the factor "alveolar bone level" (P=0.0605) did not. The thicker cement layer/post n. 3 presented bond strength values higher than those with cement layer/post n. 1. CONCLUSION: Bond strength was not influenced by different scenarios of alveolar bone resorption, although higher bond strength values were found for thinner cement layers.

Prospectus of waste stabilization ponds in Ceará, Northeast Brazil.

WSP technology has been used in Ceará, Northeast Brazil, since middle 1970s. There are presently 96 ponds plants and most of them are comprised by single cells (40%) and series of 3 ponds (35%). They were under loaded due to incomplete house connections to the sewerage network and low per capita wastewater contributions. Highest removal rates of organic material, ammonia and faecal coliform were found in 3 pond series. Faecal coliform removal was in accordance with the literature and series of ponds reached numbers ≤10(5) cells/100 ml. In series with 4 and 5 ponds FC was below 10(3) cells/100 ml. Ammonia removal varied from 30 to 80% and total phosphorus the removal was not significant. An increase in the number of maturation ponds enhances nutrient and coliform removal. Up-grading schemes should be investigated as well as effluent reuse potential.

Biaxial flexural strength of CAD/CAM ceramics.

AIM: Aim of the study was to evaluate the biaxial flexural strength of ceramics processed using the Cerec inLab system. The hypothesis was that the flexural strength would be influenced by the type of ceramic. METHODS: Ten samples (ISO 6872) of each ceramic (N.=50/n.=10) were made using Cerec inLab (software Cerec 3D) (Ø:15 mm, thickness: 1.2 mm). Three silica-based ceramics (Vita Mark II [VM], ProCad [PC] and e-max CAD ECAD]) and two yttria-stabilized tetragonal-zirconia-polycrystalline ceramics (Y-TZP) (e-max ZirCad [ZrCAD] and Vita In-Ceram 2000 YZ Cubes [VYZ]) were tested. The samples were finished with wet silicone carbide papers up to 1 200-grit and polished in a polishing machine with diamond paste (3 µm). The samples were then submitted to biaxial flexural strength testing in a universal testing machine (EMIC), 1 mm/min. The data (MPa) were analyzed using the Kruskal-Wallis and Dunn (5%) tests. Scanning electronic microscopy (SEM) was performed on a representative sample from each group. RESULTS: The values (median, mean±sd) obtained for the experimental groups were: VM (101.7, 102.1±13.65 MPa), PC (165.2, 160±34.7 MPa), ECAD (437.2, 416.1±50.1 MPa), ZrCAD (804.2, 800.8±64.47 MPa) and VYZ (792.7, 807±100.7 MPa). The type of ceramic influenced the flexural strength values (P=0.0001). The ceramics ECADa, e-max ZrCADa and VYZa presented similar flexural strength values which were significantly higher than the other groups (PCb and VM IIb), which were similar statistically between them (Dunn's test). The hypothesis was accepted. CONCLUSION: The polycrystalline ceramics (Y-TZP) should be material chosen for make FPDs because of their higher flexural strength values.

Cementation of fiber post: influence of the cement insertion techniques on the bond strength of the fiber post-root dentin and the quality of the cement layer.

AIM: The aim of this study was to assess the influence of resin cement insertion methods on the bond strength of a fiber post to root dentin and quality of the cement layer. METHODS: Forty bovine single-roots (length =16 mm) were randomly allocated into four groups, according to the cement insertion methods (N.=10): Gr1- Lentulo drill #40, Gr2- Centrix syringe, Gr3- Explorer #5, Gr4- fiber post. The root canals were prepared at 12 mm, using preparation bur # 3 of a cylinder quartz-FRC post (Aesthet post-plus, Bisco). The fiber posts were cemented using a multi-step etch-and-rinse adhesive system (All Bond 2®, Bisco) and a dual-cured resin cement (Duolink, Bisco). Each root was cut into seven samples: four samples of 1.8 mm thickness for push-out testing, and three with 0.5 mm for cement layer quality analyzing. One-way ANOVA was used for the push-out test values and the One-Way Kruskal-Wallis (P<0.05) and Dunn (10%) tests for the cement layer analyzes. RESULTS: ANOVA showed that the cement layer quality was affected by the cement insertion methods (P=0.0044): Gr1 (3.8 ± 1.3a), Gr2 (3.2 ± 1.3a), Gr3 (5.2 ± 1.5a,b) and Gr4 (5.2 ± 1.5b) (Dunn test), whereas the bond strength (MPa) was not affected by cement insertion methods: G1 (4.2 ± 1.3), G2 (3.2 ± 1.8), G3 (4.5 ± 0.9), G4 (3.1 ± 1.3). CONCLUSION: The fiber posts should be cemented with the assistance of the lentulo drill or centrix syringe to promote the best cement layer results.

Durability of feldspathic veneering ceramic on glass-infiltrated alumina ceramics after long-term thermocycling.

AIM: This study compared the bond strength durability of a feldspathic veneering ceramic to glass-infiltrated reinforced ceramics in dry and aged conditions. METHODS: Disc shaped (thickness: 4 mm, diameter: 4 mm) of glass-infiltrated alumina (In-Ceram Alumina) and glass-infiltrated alumina reinforced by zirconia (In-Ceram Zirconia) core ceramic specimens (N=48, N=12 per groups) were constructed according to the manufacturers' recommendations. Veneering ceramic (VITA VM7) was fired onto the core ceramics using a mold. The core-veneering ceramic assemblies were randomly divided into two conditions and tested either immediately after specimen preparation (Dry) or following 30000 thermocycling (5-55 ºC±1; dwell time: 30 seconds). Shear bond strength test was performed in a universal testing machine (cross-head speed: 1 mm/min). Failure modes were analyzed using optical microscope (x20). The bond strength data (MPa) were analyzed using ANOVA (α=0.05). RESULTS: Thermocycling did not decrease the bond strength results for both In-Ceram Alumina (30.6±8.2 MPa; P=0.2053) and In-Ceram zirconia (32.6±9 MPa; P=0.3987) core ceramic-feldspathic veneering ceramic combinations when compared to non-aged conditions (28.1±6.4 MPa, 29.7±7.3 MPa, respectively). There were also no significant differences between adhesion of the veneering ceramic to either In-Ceram Alumina or In-Ceram Zirconia ceramics (P=0.3289). Failure types were predominantly a mixture of adhesive failure between the veneering and the core ceramic together with cohesive fracture of the veneering ceramic. CONCLUSION: Long-term thermocycling aging conditions did not impair the adhesion of the veneering ceramic to the glass-infiltrated alumina core ceramics tested.

Silica infiltration in partially stabilized zirconia: Effect of hydrothermal aging on mechanical properties.

It aimed to evaluate if silica infiltration might influence the hydrothermal degradation of zirconia by determining: the phases formed, hardness, microstructure, and flexural strength of a 3Y-TZP. Yttria partially stabilized zirconia discs (1.2 mm thickness x 13 mm diameter) (InCeram YZ, Vita Zanhfabrik) were produced and assigned into 6 groups, considering 2 factors: silica infiltration in 2 levels (as-sintered or infiltration) and hydrothermal aging (LTD-Low Temperature Degradation) in 3 levels (baseline, aging at 132 °C for 35 h or 140 h). All the groups were subjected to the biaxial flexural test (n = 30), and Vickers hardness (n = 42). Weibull analysis was performed to determine the Weibull moduli (m) and characteristic strenghts (σ0). The specimens were characterized by scanning electron microscopy (SEM) to evaluate microstructure and X-ray diffraction (XRD) for phases percentages determination. For as-sintered condition: there was saturation of the amount of monoclinic zirconia after 35 h of hydrothermal aging, with 66% of monoclinic zirconia formed on the surface. LTD generated a progressive reduction in hardness over time; flexural strength was increased by the 35-h treatment (baseline: 974 MPa; 35 h: 1161.5 MPa), but, the 140 °C treatment was deleterious (698.5 MPa). On the other hand, the infiltrated specimens had an increase in the amount of cubic zirconia on the surface and showed 26% (35h) and 31% (140h) of monoclinic zirconia after the hydrothermal aging ; the strength was kept unaltered after LTD-35 h (935.9 MPa) and an increase was observed after LTD-140 h (1033.6 MPa); the hardness values had no statistically significant changes during the process. Thus, one can concludes that the silica infiltration can prevent the decrease in the mechanical properties due to the LTD on partially stabilized zirconia materials.

High selective leishmanicidal activity of 3-hydroxy-2-methylene-3-(4-bromophenyl)propanenitrile and analogous compounds.

Sixteen not new aromatic compounds were prepared by one-pot reaction i.e. through Baylis-Hillman reaction and were the first time evaluated against promastigote Leishmania amazonensis and infected mammalian cells. Most of the compounds were selectively more active against amastigotes than the reference drug sodium stibogluconate (Pentostam, IC(50)=44.7 microM). We found that 3-hydroxy-2-methylene-3-(4-bromophenyl) propanenitrile (13) was the most active (IC(50)=12.5 microM) and safer compound (0.0 (0.9); % macrophage LDH release), being the lead compound.

A new silica-infiltrated Y-TZP obtained by the sol-gel method.

OBJECTIVE: The aim of this study was to evaluate silica infiltration into dental zirconia (VITA In-Ceram 2000 YZ, Vita Zahnfabrik) and its effects on zirconia's surface characteristics, structural homogeneity and bonding to a resin cement. METHODS: Infiltration was performed by immersion of the pre-sintered zirconia specimens in silica sols for five days (ZIn). Negative (pure zirconia specimens, ZCon-) and positive controls (specimens kept in water for 5 days, ZCon+) were also performed. After sintering, the groups were evaluated by X-ray diffraction (XRD), grazing angle X-ray diffraction (DRXR), scanning electron microscopy (SEM), contact angle measurements, optical profilometry, biaxial flexural test and shear bonding test. Weibull analysis was used to determine the Weibull modulus (m) and characteristic strength (σ0) of all groups. RESULTS: There were no major changes in strength for the infiltrated group, and homogeneity (m) was also increased. A layer of ZrSiO4 was formed on the surface. The bond strength to resin cement was improved after zirconia infiltration, acid conditioning and the use of an MDP primer. CONCLUSION: The sol-gel method is an efficient and simple method to increase the homogeneity of zirconia. Infiltration also improved bonding to resin cement. CLINICAL SIGNIFICANCE: The performance of a zirconia infiltrated by silica gel improved in at least two ways: structural homogeneity and bonding to resin cement. The infiltration is simple to perform and can be easily managed in a prosthesis laboratory.

Surface Treatments of Zirconia to Enhance Bonding Durability.

This article reviewed the surface treatments used most often to improve adhesion between zirconia and adhesive cements, focusing on their capacity to provide long-term bonding. Traditional and new treatments for zirconia bonding were searched. Some new treatments were discussed along with topographical views of the modified zirconia. New methods, such as selective infiltration etching and the low-fusing glassy porcelain application are promising, but more research is needed.

Synthesis of 4-acyl-1H-1,2,3-triazolic nucleosides.

Two simple regiospecific methodologies based on triazolic ring construction in the course of synthesis were applied for the synthesis of 1,2,3triazolic nucleoside analogues. The cycloaddition reactions between diazomalonaldehyde and appropriate glycosylamine derivatives were rather effective, producing the desired nucleosides 11, 17 and 24. Diazotization of enamines 21a and 21b led to the corresponding triazolic ribonucleoside derivatives 22a and 22b, in good yields. Deprotection reaction of 22a, 22b and 24 was easily achieved by Lewis acid catalysis, producing the corresponding ribonucleosides 23a, 23b and 25.

Microwave-assisted extraction of green coffee oil and quantification of diterpenes by HPLC.

The microwave-assisted extraction (MAE) of 13 different green coffee beans (Coffea arabica L.) was compared to Soxhlet extraction for oil obtention. The full factorial design applied to the microwave-assisted extraction (MAE), related to time and temperature parameters, allowed to develop a powerful fast and smooth methodology (10 min at 45°C) compared to a 4h Soxhlet extraction. The quantification of cafestol and kahweol diterpenes present in the coffee oil was monitored by HPLC/UV and showed satisfactory linearity (R(2)=0.9979), precision (CV 3.7%), recovery (<93%), limit of detection (0.0130 mg/mL), and limit of quantification (0.0406 mg/mL). The space-time yield calculated on the diterpenes content for sample AT1 (Arabica green coffee) showed a six times higher value compared to the traditional Soxhlet method.

Bonding of Y-TZP to dentin: effects of Y-TZP surface conditioning, resin cement type, and aging.

PURPOSE: To evaluate the effects of two surface treatments, aging, and two resin cements on shear bond strength between dentin and yttrium-stabilized tetragonal zirconia polycrystal ceramic (Y-TZP). MATERIALS AND METHODS: Eighty human molars were embedded in acrylic resin and sectioned 3 mm below the occlusal plane. These teeth and 80 cylindrical Y-TZP specimens (height, 4 mm; diameter, 3.4 mm) were divided into eight groups (n=10) using the following factors: Y-TZP surface treatment (Vi: low-fusing porcelain [vitrification] + hydrofluoric acid etching + silanization or Si: tribochemical silicatization); cementation strategies (PF: Panavia or CC: Clearfil); and storage (nonaging or aging). Bonding surfaces of 40 Y-TZP specimens received Vi treatment, and the rest received Si treatment. Half of the ceramic-tooth assemblies were cemented with Panavia, the rest with Clearfil. Shear tests were executed using 0.4-mm-thick wire at 0.5 mm/min. Data were analyzed by three-way analysis of variance and Tukey test (α=0.05). Fractures were analyzed. RESULTS: Y-TZP surface treatments did not affect bond strength (p=0.762, Vi = Si), while resin cements (p<0.001, Panavia > Clearfil) and aging (p=0.006, nonaging > aging) showed a significant effect. Most failures were in adhesive at dentin-cement interfaces; no failure occurred between zirconia and cement. CONCLUSION: When Y-TZP ceramic is bonded to dentin, the weakest interface is that between dentin and resin cement. The resin cement/Y-TZP interface was less susceptible to failures, owing to Y-TZP surface treatments.

Microwave-assisted methanolysis of green coffee oil.

Optimisation of a microwave-assisted methanolysis was performed to obtain cafestol and kahweol directly from green coffee oil (Coffea arabica). A two-factor (the methanolysis period and temperature), three-level, factorial experimental design (3(2)) was adopted. The methanolysis procedure was performed under microwave irradiation, using closed vessel and accurate fast responding internal fibre-optic temperature probe. The effects on the responses were measured by HPLC. After 3 min of microwave irradiation (hold time) at 100°C, with 500 mg of green coffee oil, a yield higher than 99% was obtained. The yield of this reaction is 26% after 2h when working under conventional heating. The methods described in the literature lead to long reaction times, poor yields and formation of side products. The microwave-assisted technique proved to be faster, avoided undesired side products and gave better conversion, when compared to conventional heating process.