Use of the DNA Checkerboard hybridization method for detection and quantitation of Candida species in oral microbiota

The DNA Checkerboard method enables the simultaneous identification of distinct microorganisms in a large number of samples and employs up to 45 whole genomic DNA probes to gram-negative and gram-positive bacterial species present in subgingival biofilms. Collectively, they account for 55%-60% of the bacteria in subgingival biofilms. In this study, we present the DNA Checkerboard hybridization as an alternative method for the detection and quantitation of Candida species in oral cavities. Our results reveal that DNA Checkerboard is sensitive enough and constitutes a powerful and appropriate method for detecting and quantifying Candida species found in the oral cavity.

The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method

Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial. species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique. (c) 2007 Elsevier GmbH. All rights reserved.

Microbial culture and checkerboard DNA-DNA hybridization assessment of bacteria in root canals of primary teeth pre- and post-endodontic therapy with a calcium hydroxide/chlorhexidine paste

Aim. To investigate the root canal microbiota of primary teeth with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. Design. Baseline samples were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed for checkerboard DNA-DNA hybridization. Results. Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black-pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically diminished compared to initial contamination (P < 0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA-DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). Conclusion. Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing.

Vegetable Oils Deacidification by Solvent Extraction: Liquid-Liquid Equilibrium Data for Systems Containing Sunflower Seed Oil at 298.2 K

Liquid-liquid equilibrium experimental data for refined sunflower seed oil, artificially acidified with commercial oleic acid or commercial linoleic acid and a solvent (ethanol + water), were determined at 298.2 K. This set of experimental data and the experimental data from Cuevas et al.,(1) which were obtained from (283.2 to 333.2) K, for degummed sunflower seed oil-containing systems were correlated using NRTL and UNIQUAC models with temperature-dependent binary parameters. The deviation between experimental and calculated compositions presented average values of (1.13 and 1.41) % for NRTL and UNIQUAC equations, respectively, indicating that the models were able to correctly describe the behavior of compounds under different temperature and solvent hydration.

Densities and Viscosities of vegetable oils of nutritional value

Densities and viscosities of five vegetable oils (Babassu oil, Buriti oil, Brazil nut oil, macadamia oil, and grape seed oil) and of three blends of Buriti oil and soybean oil were measured as a function of temperature and correlated by empirical equations. The estimation capability of two types of predictive methodologies was tested using the measured data. The first group of methods was based on the fatty acid composition of the oils, while the other was based on their triacylglycerol composition, as a multicomponent system. In general, the six models tested presented a good representation of the physical properties considered in this work. A simple method of calculation is also proposed to predict the dynamic viscosity of methyl and ethyl ester biodiesels, based on the fatty acid composition of the original oil. Data presented in this work and the developed model can be valuable for designing processes and equipment for the edible oil industry and for biodiesel production.

Evaluation of mechanical, physical and thermal performance of cement-based tiles reinforced with vegetable fibers

The objective of this work was to analyze mechanical, physical and thermal performance of roofing tiles produced with several formulations of cement-based matrices reinforced with sisal and eucalyptus fibers. The physical properties of the tiles were more influenced by the fiber content of the composite than by the type of reinforcement. The type of the fiber was the main variable for the achievement of the best results of mechanical properties. Exposure to tropical climate has caused a severe reduction in the mechanical properties of the composites. After approximately four months of age under external weathering the toughness of the vegetable fiber-cement fell to 53-68% of the initial toughness at 28 days of age. The thermal performance showed that roofing tiles reinforced with vegetable fiber are acceptable as substitutes of asbestos-cement sheets. (c) 2006 Elsevier Ltd. All rights reserved.

Hybridization and introgression across different ploidy levels in the Neotropical orchids Epidendrum fulgens and E-puniceoluteum (Orchidaceae)

The hypothesis of gene flow between species with large differences in chromosome numbers has rarely been tested in the wild, mainly because species of different ploidy are commonly assumed to be reproductively isolated from each other because of instantaneous and strong postzygotic barriers. In this study, a broad-scale survey of molecular variation was carried out between two orchid species with different ploidy levels: Epidendrum fulgens (2n = 2x = 24 chromosomes) and Epidendrum puniceoluteum (2n = 4x = 52 chromosomes). To test the strength of their reproductive barriers, we investigated the distribution of genetic variation in sympatric and allopatric populations of these two species and conducted crossing experiments. Nuclear and plastid microsatellite loci were used to genotype 463 individuals from eight populations across the geographical range of both species along the Brazilian coastal plain. All six sympatric populations analysed presented hybrid zones, indicating that hybridization between E. fulgens and E. puniceoluteum is a common phenomenon. Bayesian assignment analysis detected the presence of F(1) and F(2) individuals and also signs of introgression, demonstrating a high potential for interspecific gene flow. Introgression occurs preferentially from E. fulgens to E. puniceoluteum. Pure parental individuals of both species display strong genotype-habitat associations, indicating that environment-dependent selection could be acting in all hybrid zones. This study suggests that hybridization and introgression are evolutionary processes playing a role in the diversification of Epidendrum and indicates the importance of investigations of hybrid zones in understanding reproductive barriers and speciation processes in Neotropical orchid species.

Chromosomal studies in four species of genus Chaunus (Bufonidae, Anura): localization of telomeric and ribosomal sequences after fluorescence in situ hybridization (FISH)

Fluorescence in situ hybridization (FISH) using telomeric and ribosomal sequences was performed in four species of toad genus Chaunus: C. ictericus, C. jimi, C. rubescens and C. schneideri. Analyses based on conventional, C-banding and Ag-NOR staining were also carried out. The four species present a 2n = 22 karyotype, composed by metacentric and submetacentric chromosomes, which were indistinguishable either after conventional staining or banding techniques. Constitutive heterochromatin was predominantly located at pericentromeric regions, and telomeric sequences (TTAGGG)(n) were restricted to the end of all chromosomes. Silver staining revealed Ag-NORs located at the short arm of pair 7, and heteromorphism in size of NOR signals was also observed. By contrast, FISH with ribosomal probes clearly demonstrated absence of any heteromorphism in size of rDNA sequences, suggesting that the difference observed after Ag-staining should be attributed to differences in chromosomal condensation and/or gene activity rather than to the number of ribosomal cistrons.

Sustainable vegetable crop supply problem

We consider an agricultural production problem, in which one must meet a known demand of crops while respecting ecologically-based production constraints. The problem is twofold: in order to meet the demand, one must determine the division of the available heterogeneous arable areas in plots and, for each plot, obtain an appropriate crop rotation schedule. Rotation plans must respect ecologically-based constraints such as the interdiction of certain crop successions, and the regular insertion of fallows and green manures. We propose a linear formulation for this problem, in which each variable is associated with a crop rotation schedule. The model may include a large number of variables and it is, therefore, solved by means of a column-generation approach. We also discuss some extensions to the model, in order to incorporate additional characteristics found in field conditions. A set of computational tests using instances based on real-world data confirms the efficacy of the proposed methodology. (C) 2009 Elsevier B.V. All rights reserved.

Network genealogy of 195-bp satellite DNA supports the superimposed hybridization hypothesis of Trypanosoma cruzi evolutionary pattern

Trypanosoma cruzi is highly diverse genetically and has been partitioned into six discrete typing units (DTUs), recently re-named T. cruzi I-VI. Although T. cruzi reproduces predominantly by binary division, accumulating evidence indicates that particular DTUs are the result of hybridization events. Two major scenarios for the origin of the hybrid lineages have been proposed. It is accepted widely that the most heterozygous TcV and TcVI DTUs are the result of genetic exchange between TcII and TcIII strains. On the other hand, the participation of a TcI parental in the current genome structure of these hybrid strains is a matter of debate. Here, sequences of the T. cruzi-specific 195-bp satellite DNA of TcI, TcII, Tat, TcV, and TcVI strains have been used for inferring network genealogies. The resulting genealogy showed a high degree of reticulation, which is consistent with more than one event of hybridization between the Tc DTUs. The data also strongly suggest that Tat is a hybrid with two distinct sets of satellite sequences, and that genetic exchange between TcI and TcII parentals occurred within the pedigree of the TcV and TcVI DTUs. Although satellite DNAs belong to the fast-evolving portion of eukaryotic genomes, in >100 satellite units of nine T. cruzi strains we found regions that display 100% identity. No DTU-specific consensus motifs were identified, inferring species-wide conservation. (C) 2010 Elsevier B.V. All rights reserved.

Amazonian Vegetable Oils and Fats: Fast Typification and Quality Control via Triacylglycerol (TAG) Profiles from Dry Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry Fingerprinting

Amazonian oils and fats display unique triacylglycerol (TAG) profiles and, because of their economic importance as renewable raw materials and use by the cosmetic and food industries, are often subject to adulteration and forgery. Representative samples of these oils (andiroba, Brazil nut, buriti, and passion fruit) and fats (cupuacu, murumuru, and ucuba) were characterized without pre-separation or derivatization via dry (solvent-free) matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Characteristic profiles of TAG were obtained for each oil and tat. Dry MALDI-TOF MS provides typification and direct and detailed information, via TAG profiles, of their variable combinations of fatty acids. A database from spectra could be developed and may be used for their fast and reliable typification, application screening, and quality control.

Effect of polyol excess on the electrical property of vegetable-polyurethane film

Freestanding castor oil-based polyurethane (PU) film was obtained using spin-coating method. The effect of polyol content was analysed by means of thermally stimulated depolarisation current and AC dielectric measurements techniques. Two relaxation peaks were observed in the temperature range of -40 to 60 degrees C for PU with different polyol contents. The presence of polyol excess provides a shift to lower temperature of the a relaxation and the decrease in the activation energy of the transition in this region might be attributed to the plasticising effect of the polyol. The peak at higher temperature is due to the Maxwell-Wagner-Sillars relaxation, which also shifts in the low temperature direction as the polyol content is increased.

Characterising vehicle emissions from the burning of biodiesel made from vegetable oil

Biodiesel manufactured from canola oil was blended with diesel and used as fuel in two diesel vehicles. This study aimed to test the emissions of diesel engines using blends of 100%, 80%, 60%, 40% , 20% biodiesel and 100% petroleum diesel, and characterise the particulate matter and gaseous emissions, with particular attention to levels of polycyclic aromatic hydrocarbons (PAHs) which are harmful to humans. A real time dust monitor was also used to monitor the continuous dust emissions during the entire testing cycle. The ECE(Euro 2) drive cycle was used for all emission tests. It was found that the particle concentration was up to 33% less when the engine burnt 100% biodiesel, compared to 100% diesel. Particle emission reduced with increased percentages of biodiesel in the fuel blends. Reductions of NOx, HC and CO were limited to about 10% when biodiesel was burned. Levels of CO2 emissions from the use of biodiesel and diesel were similar. Eighteen EPA priority PAHs were targeted, with only 6 species detected in the gaseous phase from the samples . 9 PAHs were detected in particulate phases at much lower levels than gaseous PAHs. Some marked reductions were observed for less toxic gaseous PAHs such as naphthalene when burning 100% biodiesel, but the particulate PAH emissions, which have more implications to adverse health effects, were virtually unchanged and did not show a statistically significant reduction. These findings are useful to gain an understanding of the emissions and environmental impacts of biodiesel.

An exploration of the relationships between food lifestyle and vegetable consumption

A short questionnaire was completed by 276 South Australian consumers, which examined postulated relationships between personal values, food lifestyle, demographics and their usual consumption of 24 vegetables. Principal components analyses showed that consumers' vegetable consumption could be divided into several categories, most notably salad and boiled vegetables. In multiple regression analyses different sets of values and lifestyle factors predicted intakes of overall vegetable (Rsq=27 per cent), salad (16 per cent) and boiled (27 per cent) vegetables. Path analysis revealed a complex set of pathways leading from values and personal demographics through motives, perceived food attributes and cooking skills to consumption. These partly confirmed the food lifestyle model proposed by Grunert et al. The findings show that vegetable consumption has a number of contextual and cognitive antecedents but strongly suggest that other likely predictive variables require investigation.

Two important exceptions to the relationship between energy density and fat content: food with reduced-fat claims and high-fat vegetable-based dishes

Objective: To test the hypothesis that many foods with reduced-fat (RF) claims are relatively energy-dense and that high-fat (HF) vegetable-based dishes are relatively energy-dilute.

Design: Nutrient data were collected from available foods in Melbourne supermarkets that had an RF claim and a full-fat (FF) equivalent. Nutrient analyses were also conducted on recipes for HF vegetable-based dishes that had more than 30% energy from fat but less than 10% from saturated fat. The dietary intake data (beverages removed) from the 1995 National Nutrition Survey were used for the reference relationships between energy density (ED) and percentage energy as fat and carbohydrate and percentage of water by weight.

Statistics: Linear regression modelled relationships of macronutrients and ED. Paired t-tests compared observed and predicted reductions in the ED of RF foods compared with FF equivalents.

Results: Both FF and RF foods were more energy-dense than the Australian diet and the HF vegetable-based dishes were less energy-dense. The Australian diet showed significant relationships with ED, which were positive for percentage energy as fat and negative for percentage energy as carbohydrate. There were no such relationships for the products with RF claims or for the HF vegetable-based dishes.

Conclusion: While, overall, a reduced-fat diet is relatively energy-dilute and is likely to protect against weight gain, there appear to be two important exceptions. A high intake of products with RF claims could lead to a relatively energy-dense diet and thus promote weight gain. Alternatively, a high intake of vegetable-based foods, even with substantial added fat, could reduce ED and protect against weight gain.