Isolation, characterization and biological activity of acidic phospholipase A(2) isoforms from Bothrops jararacussu snake venom

Acidic phospholipase A(2) (PLA(2)) isoforms in snake venoms, particularly those from Bothrops jararacussu, have not been characterized. This article reports the isolation and partial biochemical, functional and structural characterization of four acidic PLA(2)s (designated SIIISPIIA, SIIISPIIB, SIIISPIIIA and SIIISPIIIB) from this venom. The single chain purified proteins contained 122 amino acid residues and seven disulfide bonds with approximate molecular masses of 15 kDa and isoelectric points of 5.3. The respective N-terminal sequences were: SIIISPIIA-SLWQFGKMIDYVMGEEGAKS; SIIISPIIB-SLWQFGKMIFYTGKNEPVLS; SIIISPIIIA-SLWQFGKMILYVMGGEGVKQ and SIIISPIIIB-SLWQFGKMIFYEMTGEGVL. Crystals of the acidic protein SIIISPIIIB diffracted beyond 1.8 Angstrom resolution. These crystals are monoclinic with unit cell dimensions of a = 40.1 Angstrom, b = 54.2 Angstrom and c = 90.7 Angstrom. The crystal structure has been refined to a crystallographic residual of 16.1% (R-free = 22.9%). Specific catalytic activity (U/mg) of the isolated acidic PLA(2)s were SIIISPIIA = 290.3 U/mg; SIIISPIIB = 279.0 U/mg; SIIISPIIIA = 270.7 U/mg and SIIISPIIIB = 96.5 U/mg. Although their myotoxic activity was low, SIIISPIIA, SIIISPIIIB and SIIISPIIIA showed significant anticoagulant activity. However, there was no indirect hemolytic activity. SIIISPIIIB revealed no anticoagulant, but presented indirect hemolytic activity. With the exception of SIIISPIIIB, which inhibited platelet aggregation, all the others were capable of inducing time-independent edema. Chemical modification with 4-bromophenacyl bromide did not inhibit the induction of edema, but did suppress other activities. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

The significance of root growth on cotton nutrition in an acidic low-P soil

Toxic levels of Al and low availability of Ca have been shown to decrease root growth, which can also be affected by P availability. In the current experiment, initial plant growth and nutrition of cotton (Gossypium hirsutum var. Latifolia) were studied as related to its root growth in response to phosphorus and lime application. The experiment was conducted in Botucatu, Sao Paulo, Brazil, in pots containing a Dark Red Latosol (Acrortox, 20% clay, 72% sand). Lime was applied at 0.56, 1.12 and 1.68 g kg -1 and phosphorus was applied at 50, 100 and 150 mg kg -1. Two cotton (cv. IAC 22) plants were grown per pot for up to 42 days after plant emergence. There was no effect of liming on shoot dry weight, root dry matter yield, root surface and length, but root diameter was decreased with the increase in soil Ca. Shoot dry weight, as well as root length, surface and dry weight were increased with soil P levels up to 83 mg kg -1. Phosphorus concentration in the shoots was increased from 1.6 to 3.0 g kg -1 when soil P was increased from 14 to 34 mg kg -1. No further increases in P concentration were observed with higher P rates. The shoot/root ratio was also increased with P application as well as the amount of nutrients absorbed per unit of root surface. In low soil P soils the transport of the nutrient to the cotton root surface limits P uptake. In this case an increase in root growth rate due to P fertilisation does not compensate for the low P diffusion in the soil.

Short Communication: In vitro evaluation of the abrasiveness of acidic dentifrices.

AIM: This in vitro study evaluated the abrasiveness of acidic fluoride (F) dentifrices with different F concentrations on bovine enamel. METHODS: Enamel blocks (4.0 x 4.0 mm2, n=120) were selected according to their surface microhardness and divided into 12 groups. Slurries of dentifrices were used containing 0 (placebo), 275, 412, 550 and 1,100 ppm F (pH 4.5 or 7.0), as well as testing two commercial dentifrices (Crest, positive control, 1,100 ppm F and Colgate Baby, 500 ppm F). Enamel blocks were partially protected with an adhesive tape (control area) and then brushed by an automated toothbrushing machine (16,000 strokes). During this process, 0.4 ml of the slurries were injected every 2 mins on the enamel blocks. After toothbrushing, enamel wear was determined by profilometry. STATISTICS: Results were analyzed by ANOVA and Tukey's test (p<0.05). RESULTS: The mean values for pH in the suspensions during treatment were 6.93, 4.32, 7.56 and 8.19 for neutral experimental dentifrices, acidic experimental dentifrices, Crest and Colgate baby, respectively. The abrasiveness of the acidic dentifrices was similar (p<0.05) to the neutral ones, whereas commercial dentifrices yielded lower abrasion (p<0.05). CONCLUSION: It was concluded that a reduction of the pH of dentifrices does not increase their abrasiveness.

In vitro assessment of dentin erosion after immersion in acidic beverages: Surface profile analysis and energy-dispersive X-ray fluorescence spectrometry study

The aim of this study was to investigate the effects of some acidic drinks on dentin erosion, using methods of surface profile (SP) analysis and energy-dispersive X-ray fluorescence spectrometry (EDXRF). One hundred standardized dentin slabs obtained from bovine incisor roots were used. Dentin slabs measuring 5x5 mm were ground flat, polished and half of each specimen surface was protected with nail polish. For 60 min, the dentin surfaces were immersed in 50 mL of 5 different drinks (Gatorade®, Del Valle Mais orange juice®, Coca-Cola®, Red Bull® and white wine), 20 blocks in each drink. The pH of each beverage was measured. After the erosive challenge, the nail polish was removed and SP was analyzed. The mineral concentration of dentin surfaces was determined by means of EDXRF. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). SP analysis showed that Red-Bull had the highest erosive potential (p<0.05). EDXRF results exhibited a decrease in phosphate in the groups immersed in Red-Bull, orange juice and white wine (p<0.05), and no significant difference in calcium content between the reference surface and eroded surface. In conclusion, this study demonstrated that all studied beverages promoted erosion on root dentin and Red Bull had the highest erosive potential. There was no correlation between pH of beverages and their erosive potential and only the P content changed after erosive challenge.

Effect of low fluoride acidic dentifrices on dental remineralization

This study evaluated the capacity of fluoride acidic dentifrices (pH 4.5) to promote enamel remineralization using a pH cycling model, comparing them with a standard dentifrice (1,100 μgF/g). Enamel blocks had their surface polished and surface hardness determined (SH). Next, they were submitted to subsurface enamel demineralization and to postdemineralization surface hardness analysis. The blocks were divided into 6 experimental groups (n=10): placebo (without F, pH 4.5, negative control), 275, 412, 550, 1,100 μgF/g and a standard dentifrice (positive control). The blocks were submitted to pH cycling for 6 days and treatment with dentifrice slurries twice a day. After pH cycling, surface and crosssectional hardness were assessed to obtain the percentage of surface hardness recovery (%SHR) and the integrated loss of subsurface hardness (δKHN). The results showed that %SHR was similar among acidic dentifrices with 412, 550, 1,100 μgF/g and to the positive control (Tukey's test; p>0.05). For ΔKHN, the acidic dentifrice with 550 μg F/g showed a better performance when compared with the positive control. It can be concluded that acidic dentifrice 550 μgF/g had similar remineralization capacity to that of positive control.

Role of acidic and basic electrolytes on the structure and morphology of cathodically reduced indium tin oxide (ITO) substrates

In this report we track the structural changes suffered by ITO along galvanostatic polarization at different current densities by X-ray diffraction and SEM micrographs. The XRD shown that cathodic treatment induces structural change in ITO, characterized by appearing peaks set distinct from ITO original structure associated to metallic phase of the solid solution of In-Sn. It is interesting to note that although the different ions present in the solution are not, at least to a noticeable degree, incorporated in the metallic phase, the SEM images show that they do influence its formation, pointing to some type of adsorptive mechanism of the inert ions during the lateral diffusion of the metallic ions. © 2013 Elsevier Ltd. All rights reserved.

Mechanisms of action of fluoridated acidic liquid dentifrices against dental caries

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

Electrochemical behaviour of (N-R-4-cyanopyridinium)pentaammineruthenium(II) derivatives in acidic medium. Hydrolysis of coordinated nitriles

The electrochemical behaviour of N-R-4-cyanopyridinium (4-rcp) (R = methyl, decyl, dodecyl, or benzyl) coordinated to pentaammineruthenium(II) in CF3COOH-CF3COONa (μ = 0.1 M, pH 3) aqueous medium was studied by means of cyclic voltammetry and constant potential electrolysis. The electrochemical oxidation of the metallic centre (Ep ca 0.51 V/SCE) can be described as a reversible monoelectronic charge-transfer followed by an irreversible chemical reaction, which is the hydrolysis of N-R-4-cyanopyridiniumpentaammineruthenium(III) (A) to N-R-4-carboxamidepyridiniumruthenium (III) (B) with the kf1 values depending on the type of alkyl group. The E 1 2 values are not significantly influenced by the nature of the alkyl group. At more negative potential (ca -0.5 V/SCE), B undergoes an electrochemical reduction followed by an aquation reaction to produce aquopentaammineruthenium(II) and free N-R-4-carboxamidepyridinium. The amide was identified by comparison of its cyclic voltammogram and UV-vis spectrum with that of a sample prepared by chemical reaction. The results were also discussed by comparison with other systems, and show that nitrile-amide conversion catalysed by pentaammineruthenium(II) complexes is possible. © 1994.

Structural and Functional Insights from the Metagenome of an Acidic Hot Spring Microbial Planktonic Community in the Colombian Andes

A taxonomic and annotated functional description of microbial life was deduced from 53 Mb of metagenomic sequence retrieved from a planktonic fraction of the Neotropical high Andean (3,973 meters above sea level) acidic hot spring El Coquito (EC). A classification of unassembled metagenomic reads using different databases showed a high proportion of Gammaproteobacteria and Alphaproteobacteria (in total read affiliation), and through taxonomic affiliation of 16S rRNA gene fragments we observed the presence of Proteobacteria, micro-algae chloroplast and Firmicutes. Reads mapped against the genomes Acidiphilium cryptum JF-5, Legionella pneumophila str. Corby and Acidithiobacillus caldus revealed the presence of transposase-like sequences, potentially involved in horizontal gene transfer. Functional annotation and hierarchical comparison with different datasets obtained by pyrosequencing in different ecosystems showed that the microbial community also contained extensive DNA repair systems, possibly to cope with ultraviolet radiation at such high altitudes. Analysis of genes involved in the nitrogen cycle indicated the presence of dissimilatory nitrate reduction to N2 (narGHI, nirS, norBCDQ and nosZ), associated with Proteobacteria-like sequences. Genes involved in the sulfur cycle (cysDN, cysNC and aprA) indicated adenylsulfate and sulfite production that were affiliated to several bacterial species. In summary, metagenomic sequence data provided insight regarding the structure and possible functions of this hot spring microbial community, describing some groups potentially involved in the nitrogen and sulfur cycling in this environment. Citation: Jimenez DJ, Andreote FD, Chaves D, Montana JS, Osorio-Forero C, et al. (2012) Structural and Functional Insights from the Metagenome of an Acidic Hot Spring Microbial Planktonic Community in the Colombian Andes. PLoS ONE 7(12): e52069. doi:10.1371/journal.pone.0052069

Solid-phase microextraction combined with comprehensive two-dimensional gas chromatography for fatty acid profiling of cell wall phospholipids

The combination of solid-phase microextraction (SPME) with comprehensive two-dimensional gas chromatography is evaluated here for fatty acid (FA) profiling of the glycerophospholipid fraction from human buccal mucosal cells. A base-catalyzed derivatization reaction selective for polar lipids such as glycerophospholipid was adopted. SPME is compared to a miniaturized liquidliquid extraction procedure for the isolation of FA methyl esters produced in the derivatization step. The limits of detection and limits of quantitation were calculated for each sample preparation method. Because of its lower values of limits of detection and quantitation, SPME was adopted. The extracted analytes were separated, detected, and quantified by comprehensive two-dimensional gas chromatography with flame ionization detection (FID). The combination of SPME and comprehensive two-dimensional gas chromatography with FID, using a selective derivatization reaction in the preliminary steps, proved to be a simple and fast procedure for FA profiling, and was successfully applied to the analysis of adult human buccal mucosal cells.

Synthetic phosphoethanolamine a precursor of membrane phospholipids reduce tumor growth in mice bearing melanoma B16-F10 and in vitro induce apoptosis and arrest in G2/M phase

Phosphoethanolamine (Pho-s) is a compound involved in phospholipid turnover, acting as a substrate for many phospholipids of the cell membranes, especially phosphatidylcholine. We recently reported that synthetic Pho-s has potent effects on a wide variety of tumor cells. To determine if Pho-s has a potential antitumor activity, in this study we evaluated the activity of Pho-s against the B16-F10 melanoma both in vitro and in mice bearing a dorsal tumor. The treatment of B16F10 cells with Pho-s resulted in a dose-dependent inhibition of cell proliferation. At low concentrations, this activity appears to be involved in the arrest of the cell cycle at G2/M, while at high concentrations Pho-s induces apoptosis. In accordance with these results, the loss of mitochondrial potential and increased caspase-3 activity suggest that Phos has dual antitumor effects; i.e. it induces apoptosis at high concentrations and modulates the cell cycle at lower concentrations. In vivo, we evaluated the effect of Pho-s in mice bearing B16-F10 melanoma. The results show that Pho-s reduces the tumoral volume increasing survival rate. Furthermore, the tumor doubling time and tumor delays were substantially reduced when compared with untreated mice. Histological analyses reveal that Pho-s induces changes in cell morphology, typical characteristics of apoptosis, in addition the large areas of necrosis correlating with a reduction of tumor size. The results presented here support the hypothesis that Pho-s has antitumor effects by the induction of apoptosis as well as the inhibition of cell proliferation by arrest at G2/M. Thus, Pho-s can be regarded as a promising agent for the treatment of melanoma. Published by Elsevier Masson SAS.

Aniline-1,4-benzoquinone as a model system for the characterization of products from aniline oligomerization in low acidic media

Increase in the pH medium of aniline polymerization is used for giving products of different morphologies, which are often wrongly attributed to PANI chains. Infrared and Raman spectroscopic data, supported by quantum chemical calculations, show that aniline-1,4-benzoquinone (AnBzq) is a model system for the characterization of the products of aniline oligomerization in low acidic media. The Raman spectra excited at different laser lines reveal the bichromophoric nature of AnBzq, whose absorption bands at 550 nm and 440 nm can be attributed to pi-pi* transitions of the delocalized benzoquinone and amino-phenyl moieties, respectively. (C) 2012 Elsevier B.V. All rights reserved.

The influence of the Pt crystalline surface orientation on the glycerol electro-oxidation in acidic media

We investigated the electrochemical oxidation of glycerol on low-index Pt single crystals in acidic media (H2SO4 and HClO4) by cyclic voltammetry and Fourier Transform Infrared (FTIR) spectroscopy and we verified that this is a surface sensitive reaction. Pt(100) and Pt(110) surface structures favor the breaking of the C-C-C bond at low potentials (say 0.05 V), as seen by the formation of CO, one of the adsorbed residues of the glycerol dissociation, which poisons these surfaces even at high potentials. Pt(111) surface structure does not favor the C-C-C bond breaking at potentials as low as 0.05 V. However, Pt(111) is less poisoned by residues of glycerol dissociation and, for this reason, it is more active for glycerol oxidation than Pt(100) and Pt(110) at low potentials. Carbonyl containing compounds and CO2 were detected as reaction products of the glycerol oxidation on all investigated single-crystal Pt surfaces. The ratio between CO2 and carbonyl containing compounds is clearly much higher for Pt(100) and Pt(110) than for Pt(111). (C) 2012 Elsevier Ltd. All rights reserved.

OSCILLATORY DYNAMICS IN SYSTEMS CONTAINING BROMATE AND 1,4-CYCLOHEXANEDIONE IN ACIDIC MEDIA. I. THE EFFECT OF TEMPERATURE.

OSCILLATORY DYNAMICS IN SYSTEMS CONTAINING BROMATE AND 1,4-CYCLOHEXANEDIONE IN ACIDIC MEDIA. I. THE EFFECT OF TEMPERATURE. We present in this work the influence of temperature on the dynamics of homogeneous chemical systems containing bromate and 1,4-cyclohexanedione (1,4-CHD) in acidic media. In particular, the following systems were studied: bromate/1,4-CHD/acid, bromate/1,4-CHD/ferroin/acid and bromate/1,4-CHD/trisbipyridine ruthenium/acid. Investigations were carried out by means of an electrochemical probe, at five temperatures between 5 and 45 degrees C. Activation energies (E-a) were estimated in different ways for the pre-oscillatory and oscillatory regimes. In any case, the E-a was found to depend on the catalyst, composition and initial concentrations. In addition, it was observed that ferroin and trisbipyridine ruthenium act as catalysts only during the transition between the induction period and oscillatory regime.

Effect of low fluoride acidic dentifrices on dental remineralization

This study evaluated the capacity of fluoride acidic dentifrices (pH 4.5) to promote enamel remineralization using a pH cycling model, comparing them with a standard dentifrice (1,100 µgF/g). Enamel blocks had their surface polished and surface hardness determined (SH). Next, they were submitted to subsurface enamel demineralization and to post-demineralization surface hardness analysis. The blocks were divided into 6 experimental groups (n=10): placebo (without F, pH 4.5, negative control), 275, 412, 550, 1,100 µgF/g and a standard dentifrice (positive control). The blocks were submitted to pH cycling for 6 days and treatment with dentifrice slurries twice a day. After pH cycling, surface and cross-sectional hardness were assessed to obtain the percentage of surface hardness recovery (%SHR) and the integrated loss of subsurface hardness (ΔKHN). The results showed that %SHR was similar among acidic dentifrices with 412, 550, 1,100 µgF/g and to the positive control (Tukey's test; p>0.05). For ΔKHN, the acidic dentifrice with 550 µg F/g showed a better performance when compared with the positive control. It can be concluded that acidic dentifrice 550 µgF/g had similar remineralization capacity to that of positive control.