Physico-chemical properties of MTA and a novel experimental cement

Aim: To evaluate the release of calcium ions, pH and conductivity of a new experimental dental cement (EC) and to compare them with those of mineral trioxide aggregate (MTA-Angelus). Methodology: Five samples of each cement were prepared using plastic tubes 1 mm in diameter and 10 mm long. Each sample was sealed in a test tube containing 10 mL deionized water which was analysed after 24, 48, 72, 96, 192, 240 and 360 h for pH, electrical conductivity and calcium release. The concentration of calcium ions was obtained through atomic absorption spectroscopy technique. The data were analysed statistically using the analysis of variance (ANOVA) and the Student's test (t-test). Results: The pH of the storage solutions was not affected by the material and the interaction of material with time (P > 0.05). However, the time of immersion was significant (P < 0.01) for both materials. For the electric conductivity and calcium release, the interaction of material with time was statistically significant (P < 0.01), indicating that EC and MTA-Angelus did not behave in a similar manner. Conclusions: The experimental cement released calcium and increased the pH of the storage solutions in a similar manner to MTA-Angelus. However, EC showed significantly higher calcium release than commercial MTA-Angelus after 24 h. © 2005 International Endodontic Journal.

A new firefly luciferase with bimodal spectrum: Identification of structural determinants of spectral pH-sensitivity in firefly luciferases

Fireflies emit flashes in the green-yellow region of the spectrum for the purpose of sexual attraction. The bioluminescence color is determined by the luciferases. It is well known that the in vitro bioluminescence color of firefly luciferases can be shifted toward the red by lower pH and higher temperature; for this reason they are classified as pH-sensitive luciferases. However, the mechanism and structural origin of pH sensitivity in fireflies remains unknown. Here we report the cloning of a new luciferase from the Brazilian twilight active firefly Macrolampis sp2, which displays an unusual bimodal spectrum. The recombinant luciferase displays a sensitive spectrum with the peak at 569 nm and a shoulder in the red region. Comparison of the bioluminescence spectra of Macrolampis, Photinus and Cratomorphus firefly luciferases shows that the distinct colors are determined by the ratio between green and red emitters under luciferase influence. Comparison of Macrolampis luciferase with the highly similar North American Photinus pyralis luciferase (91%) showed few substitutions potentially involved with the higher spectral sensitivity in Macrolampis luciferase. Site-directed mutagenesis showed that the natural substitution E354N determines the appearance of the shoulder in the red region of Macrolampis luciferase bioluminescence spectrum, helping to identify important interactions and residues involved in the pH-sensing mechanism in firefly luciferases. © 2005 American Society for Photobiology.

An ultrafiltration membrane, prepared with MSU-type mesoporous silica: Preparation and specific filtration behavior

We report preparation and the singular filtration properties of an ultrafiltration membrane made with MSU-type mesoporous silica that exhibits cylindrical pores aligned mostly normal to the support. This membrane supported on tubular commercial macroporous alumina supports was prepared by the interfacial growth mechanism between stable silica-surfactant hybrid micelles made of the association of silica oligomers with polyethyleneoxide-based (PEO) surfactants and sodium fluoride, a well-known silica condensation catalyst. It appears that the combined effect of the silica nature of the membrane, whose surface charge can be easily adjusted by changing the pH and the non-connected cylindrical shape of the pores provides a new behavior in the retention properties, as proved by the filtration of polyoxyethylene polymers (PEO) with different molecular weights. Depending on the filtration conditions, a rejection rate of 80 % and a steep cut-off at 2,000 Da can be obtained or, on the reverse, polymers three times bigger than the pore diameter can diffuse through the membrane. This new filtration mechanism, which opens up new modes of separation modes, is explained in the light of both topology of the porous network and pH-dependent interactions between PEO polymers and silica porous media. © 2005 Elsevier B.V. All rights reserved.

Bovine serum albumin displays luciferase-like activity in presence of luciferyl adenylate: Insights on the origin of protoluciferase activity and bioluminescence colours

Luciferyl adenylate, the key intermediate in beetle bioluminescence, is produced through adenylation of D-luciferin by beetle luciferases and also by mealworm luciferase-like enzymes which produce a weak red chemiluminescence. However, luciferyl adenylate is only weakly chemiluminescent in water at physiological pH and it is unclear how efficient bioluminescence evolved from its weak chemiluminescent properties. We found that bovine serum albumin (BSA) and neutral detergents enhance luciferyl adenylate chemiluminescence by three orders of magnitude, simulating the mealworm luciferase-like enzyme chemiluminescence properties. These results suggest that the beetle protoluciferase activity arose as an enhanced luciferyl adenylate chemiluminescence in the protein environment of the ancestral AMP-ligase. The predominance of luciferyl adenylate chemiluminescence in the red region under most conditions suggests that red luminescence is a more primitive condition that characterized the original stages of protobioluminescence, whereas yellow-green bioluminescence may have evolved later through the development of a more structured and hydrophobic active site. Copyright © 2006 John Wiley & Sons, Ltd.

Photodegradation of soluble and emulsive cutting fluids using TiO 2 as catalyst

This paper describes results of the photo-degradation of three types of soluble and emulsive cutting fluids in an aqueous medium, using TiO2 as catalyst in suspension and UV radiation. The TiO2 proved to be an effective catalyst for the degradation of the cutting fluids investigated. The degradation rate depends on pH and nature of the fluids. The best performance of catalyst was observed at pH 8.0 for all the fluids when most of 70% of the organic load was decomposed. ©2006 Sociedade Brasileira de Química.

Multiparticulate systems of pectin-chitosan: Study of swelling and drug release

The aim of this study was to prepare multiparticulate systems of pectin:chitosan (PC:CS) and to evaluate their swelling ratio and the drug release in different environments. PC:CS particles containing triamcinolone were prepared by a complex coacervation/ionotropic gelation method in aqueous environment. The polymer ratio, the calcium concentration and the contact time of the capsules with chitosan dispersion for particles formation and the structures obtained were analyzed. The systems were characterized in relation to morphology, size, swelling, and drug release behavior. The methodology used allowed the production of spherical particles with narrow range of size distribution. The entrapment efficiency for triamcinolone was 84.31 ± 439. It was observed that the particles present a relatively low swelling ratio in acidic medium and a larger swelling ratio in enteric medium. The release profile was dependent on pH and can be related with the swelling ratio.

Chloroquine inhibits Paracoccidioides brasiliensis survival within human monocytes by limiting the availability of intracellular iron

The mechanisms used by Paracoccidioides brasiliensis (Pb 18) to survive into monocytes are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens, including P. brasiliensis, whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Chloroquine, by virtue of its basic properties, has been shown to prevent release of iron from holotransferrin by raising endocytic and lysosomal pH, and thereby interfering with normal iron metabolism. Then, in view of this, we have studied the effects of CHLOR on P. brasiliensis multiplication in human monocytes and its effect on the murine paracoccidioidomycosis. CHLOR induced human monocytes to kill P. brasiliensis. The effect of CHLOR was reversed by FeNTA, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment. CHLOR treatment of Pb 18-infected BALB/c mice significantly reduced the viable fungi recovery from lungs, during three different periods of evaluation, in a dose-dependent manner. This study demonstrates that iron is of critical importance to the survival of P. brasiliensis yeasts within human monocytes and the CHLOR treatment in vitro induces Pb 18 yeast-killing by monocytes by restricting the availability of intracellular iron. Besides, the CHLOR treatment in vivo significantly reduces the number of organisms in the lungs of Pb-infected mice protecting them from several infections. Thus, CHLOR was effective in the treatment of murine paracoccidioidomycosis, suggesting the potential use of this drug in patients' treatment.

Dinâmica da regeneração do subsolo de áreas degradadas dentro do bioma Cerrado

The objective of this research was to evaluate the physical and chemical conditions of revegetated subsoil in degraded areas in a Cerrado biome and to verify which plants promote better conditions for soil recovery. The research was conducted in the remaining area of the hydroelectrical plant site at Ilha Solteira (SP). The experimental design was a completely randomized with five treatments and three replications. The treatments consisted of: natural regeneration area, brachiaria area, Pinus sp. area, exposed soil area and Cerrado grassland (used as control). The following soil characteristics were appraised: porosity; density, stability of aggregates, infiltration rate, temperature, exchangeable cations, organic matter, pH and potential acidity. The results show that Pinus is not a good species to recover the chemical attributes of the subsoil. The revegetated areas need to be improved in their physical attributes to allow a better development of the vegetation. Brachiaria and the natural regeneration were the most promising treatments, presenting results similar to natural Cerrado.

Production of cellulolytic and hemicellulolytic enzymes from Aureobasidium pulluans on solid state fermentation

This article investigates a strain of the yeast Aureobasidium pullulans for cellulase and hemicellulase production in solid state fermentation. Among the substrates analyzed, the wheat bran culture presented the highest enzymatic production (1.05 U/mL endoglucanase, 1.3 U/mL β-glucosidase, and 5.0 U/mL xylanase). Avicelase activity was not detected. The optimum pH and temperature for xylanase, endoglucanase and β-glucosidase were 5.0 and 50, 4.5 and 60, 4.0 and 75°C, respectively. These enzymes remained stable between a wide range of pH. The β-glucosidase was the most thermostable enzyme, remaining 100% active when incubated at 75°C for 1 h. © 2007 Humana Press Inc.

Isolation and growth of the symbiotic fungus of Atta capiguara (Hymenoptera: Formicidae)

This work's objectives were to isolate and evaluate the growth of the symbiotic fungus of Atta capiguara Gonçalves on artificial medium, under different pH and temperature conditions. Isolation was accomplished using the following media: Sabouraud, oat-agar, PDA, and PDA with the addition of extracts from the grasses Paspalum sp. Flügge and Hyparrhenia rufa (Nees) Stapf.. The medium used in the growth study was PDA with the addition of a Paspalum sp. (0.22%, w/v) extract at initial pH values of 4.5, 6.0, and 7.5. Mycelium disks were transferred to plates containing the culture medium. The plates were maintained at temperatures of 20, 23, and 26 ± 1°C. Mycelial radial growth evaluations were performed at 7, 14, 21, 28, and 35 days of incubation. Fungus isolation was obtained in all media studied. The highest radial means were obtained at initial pH values of 6.0 and 7.5 and temperatures of 23 and 26± 1°C. Greater plot losses occurred at the initial pH condition of 7.5. In general, A. capiguara fungi can be grown in the medium studied, at an initial pH of 6.0 and temperatures of 23 or 26± 1°C. Radial growth evaluations at 14 and 28 days of incubation can be recommended for substrate studies involving the symbiotic fungus.

The influence of the loop between residues 223-235 in beetle luciferase bioluminescence spectra: A solvent gate for the active site of pH-sensitive luciferases

Beetle luciferases emit a wide range of bioluminescence colors, ranging from green to red. Firefly luciferases can shift the spectrum to red in response to pH and temperature changes, whereas click beetle and railroadworm luciferases do not. Despite many studies on firefly luciferases, the origin of pH-sensitivity is far from being understood. Through comparative site-directed mutagenesis and modeling studies, using the pH-sensitive luciferases (Macrolampis and Cratomorphus distinctus fireflies) and the pH-insensitive luciferases (Pyrearinus termitilluminans, Phrixotrix viviani and Phrixotrix hirtus) cloned by our group, here we show that substitutions dramatically affecting bioluminescence colors in both groups of luciferases are clustered in the loop between residues 223-235 (Photinus pyralis sequence). The substitutions at positions 227, 228 and 229 (P. pyralis sequence) cause dramatic redshift and temporal shift in both groups of luciferases, indicating their involvement in labile interactions. Modeling studies showed that the residues Y227 and N229 are buried in the protein core, fixing the loop to other structural elements participating at the bottom of the luciferin binding site. Changes in pH and temperature (in firefly luciferases), as well as point mutations in this loop, may disrupt the interactions of these structural elements exposing the active site and modulating bioluminescence colors. © 2007 The Authors.

Fig tree orchard formation using organic fertilization

The present work aimed at evaluating the effects of cattle manure fertilization on the growth and yield of fig trees. The cultivar 'Roxo de Valinhos' was used. One-, two-, three-and four-year-old trees were treated with cattle manure containing 0, 25, 50, 75, 100, 125 and 150% of the recommended nitrogen level per plant. The experiment was carried out using randomized blocks with 7 treatments, 5 replicates and 5 plants per experimental plot. The evaluated characteristics were: plant height, stem diameter, secondary branch length and diameter, yield per plant (kg plant-1), besides some quality characteristics of fruits such as pH, titratable acidity, soluble solids and texture. Manure application enhanced plant growth and fruit production. Significant differences were observed only for soluble solids content, pH and texture, which varied according to the crop cycle. After four crop cycles (2002/03, 2003/04, 2004/05 and 2005/06), the best results (about 5.0 kg of fruits per plant) were obtained with 100% of the recommended nitrogen dose, which corresponded to 14.3 kg of cattle manure per plant, in the last crop cycle (2005/2006).

Easiness of use and validity testing of VS-SENSE device for detection of abnormal vaginal flora and bacterial vaginosis

Accessing vaginal pH is fundamental during gynaecological visit for the detection of abnormal vaginal flora (AVF), but use of pH strips may be time-consuming and difficult to interpret. The aim of this study was to evaluate the VS-SENSE test (Common Sense Ltd, Caesarea, Israel) as a tool for the diagnosis of AVF and its correlation with abnormal pH and bacterial vaginosis (BV). The study population consisted of 45 women with vaginal pH ≥ 4.5 and 45 women with normal pH. Vaginal samples were evaluated by VS-SENSE test, microscopy and microbiologic cultures. Comparing with pH strips results, VS-SENSE test specificity was 97.8% and sensitivity of 91%. All severe cases of BV and aerobic vaginitis (AV) were detected by the test. Only one case with normal pH had an unclear result. Concluding, VS-SENSE test is easy to perform, and it correlates with increased pH, AVF, and the severe cases of BV and AV. Copyright © 2010 Gilbert G. G. Donders et al.

Production of pectate lyase by penicillium viridicatum RFC3 in solid-state and submerged fermentation

Pectate lyase (PL) was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w), or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w), and in a submerged liquid culture with orange bagasse and wheat bran (3%) as the carbon source. PL production was highest (1,500U mL -1 or 300Ug -1 of substrate) in solid-state fermentation (SSF) on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF) was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme fromSSF and designated PL I, II, III, IV, V, andVI, while five peaks were obtained fromcrude enzyme fromSmF and labeled PL I', II', III', IV', and VII'. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35°C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45°C. Crude enzyme from SmF and PL III' showed thermophilic profiles of activity, with maximum activity at 60 and 55°C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0-10.0 and PL III was most stable in the pH range 4.0-7.0. Crude enzyme from SmF retained 70%-80% of its maximum activity in the acid-neutral pH range (4.0-7.0), but PIII showed high stability at alkaline pH (7.5-9.5). PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55°C. The differing behavior of the enzymes with respect to pH and temperature suggests that they are different isozymes. Copyright © 2010 Viviani Ferreira et al.

Effect of calcium hydroxide on pH changes of the external medium after intracoronal bleaching

Aim: This in vitro study evaluated the effect of calcium hydroxide on pH changes of the external medium after intracoronal bleaching. Materials and methods: A total of 50 extracted human premolars were prepared and filled with gutta-percha and endodontic sealer. The teeth were randomly divided into five groups according to the bleaching agents employed: (a) Sterile cotton pellet with distilled water (control group); (b) sodium perborate and distilled water; (c) sodium perborate and 10% carbamide peroxide; (d) sodium perborate and 35% hydrogen peroxide; (e) 35% hydrogen peroxide. The teeth were stored in vials containing distilled water and the pH values of the medium surrounding the teeth were analyzed. After 7-day storage, the bleaching agent was removed and replaced by calcium hydroxide, and the distilled water was changed, in which the teeth were kept stored for further 14 days. Measurement of pH of the external medium (distilled water) was performed 7 days after insertion of the bleaching agents, immediately, 7 and 14 days after insertion of the calcium hydroxide. Data were submitted to statistical analysis by the two-way ANOVA and Tukey,s test. Results: There were pH changes of the external medium at 7-day period after bleaching procedures. These results confirmed the diffusion of bleaching agents to the external medium. Conclusion: Calcium hydroxide increased the external medium pH and was effective for pH alkalinization after intracoronal bleaching. Clinical significance: Intracoronal bleaching of endodontically treated teeth may cause cervical root resorption. A possible explanation for this process is the passage of bleaching agents to the periodontal tissues yielding an inflammatory process. In an attempt to keep the neutrality of the periodontal pH, the calcium hydroxide has been recommended.Results of this study showed that this material should be always used after intracoronal bleaching.