Evaluation of Environmental Mycobacteria Contamination in a Specific Pathogen Free Animal Facility from a Tropical Country

P>With the evidence showing the protection variability of bacille Calmette-Guerin, new potential vaccines for tuberculosis have been tested around the world. One of the general concerns in tuberculosis vaccine development is the possibility of priming the host immune system with prior exposure to environmental mycobacteria antigens, which can change the efficacy of subsequent vaccination. As there is a great homology between the species from Mycobacterium genera, the previous contact of experimental animals with environmental mycobacteria could sensitize the mice and, in this way, could influence subsequent vaccine research. The aim of our study was to investigate critical points in an animal facility to search for environmental mycobacteria that eventually could be in direct or indirect contact with the experimental animals. Samples were collected from surfaces of walls, floor, animal cages and shelves and analysed using the Ogawa-Kudoh decontamination method. Samples of drinking water, food and sawdust were collected for analysis by the NALC/NaOH decontamination method. Also, the samples were cultivated directly in broth medium, without any method for decontamination. After decontamination methods, we observed bacterial colony growth in 4.31% of the total of samples analysed. These samples were stained with Ziehl-Neelsen and we did not detect any acid-fast bacilli, suggesting that the animal facility analysed is free from contamination by environmental mycobacteria and is not a source of mycobacterial antigens. Furthermore, our study showed a new paradigm in tuberculosis vaccine development: concern about the animal facility environment in terms of immune system priming of experimental animals by nascent bacterial contaminants.

TP53 and EGFR mutations in combination with lifestyle risk factors in tumours of the upper aerodigestive tract from South America

Cancers of the upper aerodigestive tract [(UADT): oral cavity, pharynx, larynx and oesophagus] have high incidence rates in some parts of South America. Alterations in the TP53 gene are common in these cancers. In our study, we have estimated the prevalence and patterns of TP53 mutations (exons 4-10) in 236 UADT tumours from South America in relation to lifestyle risk factors, such as tobacco smoking and alcohol drinking. Moreover, we have conducted a pilot study of EGFR mutations (exons 18-21) in 45 tumours from the same population. TP53 mutation prevalence was high: 59% of tumours were found to carry mutant TP53. We found an association between TP53 mutations and tobacco smoking and alcohol drinking. The mutation rate increased from 38% in never-smokers to 66% in current smokers (P-value for trend = 0.09). G:C > T:A transversions were found only in smokers (15%). Alcohol drinkers carried more G:C > A:T transitions (P = 0.08). Non-exposed individuals were more probable to carry G:C > A:T transitions at CpG sites (P = 0.01 for never-smokers and P < 0.001 for never-drinkers). EGFR mutations were found in 4% of cases. Inactivation of TP53 by mutations is a crucial molecular event in the UADT carcinogenesis and it is closely related to exposure to lifestyle risk factors. EGFR mutations do not appear to be a common event in UADT carcinogenesis in this population.

An examination of male and female odds ratios by BMI, cigarette smoking, and alcohol consumption for cancers of the oral cavity, pharynx, and larynx in pooled data from 15 case-control studies

Greater tobacco smoking and alcohol consumption and lower body mass index (BMI) increase odds ratios (OR) for oral cavity, oropharyngeal, hypopharyngeal, and laryngeal cancers; however, there are no comprehensive sex-specific comparisons of ORs for these factors. We analyzed 2,441 oral cavity (925 women and 1,516 men), 2,297 oropharynx (564 women and 1,733 men), 508 hypopharynx (96 women and 412 men), and 1,740 larynx (237 women and 1,503 men) cases from the INHANCE consortium of 15 head and neck cancer case-control studies. Controls numbered from 7,604 to 13,829 subjects, depending on analysis. Analyses fitted linear-exponential excess ORs models. ORs were increased in underweight (< 18.5 BMI) relative to normal weight (18.5-24.9) and reduced in overweight and obese categories (a parts per thousand yen25 BMI) for all sites and were homogeneous by sex. ORs by smoking and drinking in women compared with men were significantly greater for oropharyngeal cancer (p < 0.01 for both factors), suggestive for hypopharyngeal cancer (p = 0.05 and p = 0.06, respectively), but homogeneous for oral cavity (p = 0.56 and p = 0.64) and laryngeal (p = 0.18 and p = 0.72) cancers. The extent that OR modifications of smoking and drinking by sex for oropharyngeal and, possibly, hypopharyngeal cancers represent true associations, or derive from unmeasured confounders or unobserved sex-related disease subtypes (e.g., human papillomavirus-positive oropharyngeal cancer) remains to be clarified.

Cessation of alcohol drinking, tobacco smoking and the reversal of head and neck cancer risk

Background Quitting tobacco or alcohol use has been reported to reduce the head and neck cancer risk in previous studies. However, it is unclear how many years must pass following cessation of these habits before the risk is reduced, and whether the risk ultimately declines to the level of never smokers or never drinkers. Methods We pooled individual-level data from case-control studies in the International Head and Neck Cancer Epidemiology Consortium. Data were available from 13 studies on drinking cessation (9167 cases and 12 593 controls), and from 17 studies on smoking cessation (12 040 cases and 16 884 controls). We estimated the effect of quitting smoking and drinking on the risk of head and neck cancer and its subsites, by calculating odds ratios (ORs) using logistic regression models. Results Quitting tobacco smoking for 1-4 years resulted in a head and neck cancer risk reduction [OR 0.70, confidence interval (CI) 0.61-0.81 compared with current smoking], with the risk reduction due to smoking cessation after >= 20 years (OR 0.23, CI 0.18-0.31), reaching the level of never smokers. For alcohol use, a beneficial effect on the risk of head and neck cancer was only observed after >= 20 years of quitting (OR 0.60, CI 0.40-0.89 compared with current drinking), reaching the level of never drinkers. Conclusions Our results support that cessation of tobacco smoking and cessation of alcohol drinking protect against the development of head and neck cancer.

Analysis of marine bivalve shellfish from the fish market in Santos city, Sao Paulo state, Brazil, for Toxoplasma gondii

The aim of this study was to determine if Toxoplasma gondii are present in oysters (Crassostrea rhizophorae) and mussels (Mytella guyanensis) under natural conditions using a bioassay in mice and molecular detection methods. We first compared two standard protocols for DNA extraction, phenol-chloroform (PC) and guanidine-thiocyanate (GT), for both molluscs. A total of 300 oysters and 300 mussels were then acquired from the fish market in Santos city, Sao Paulo state, Brazil, between March and August of 2008 and divided into 60 groups of 5 oysters and 20 groups of 15 mussels. To isolate the parasite, five mice were orally inoculated with sieved tissue homogenates from each group of oysters or mussels. For molecular detection of T. gondii, DNA from mussels was extracted using the PC method and DNA from oysters was extracted using the GT method. A nested-PCR (Polymerase Chain Reaction) based on the amplification of a 155 bp fragment from the B1 gene of T. gondii was then performed. Eleven PCR-RFLP (Restriction Fragment Length Polymorphism) markers, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, CS3 and Apico, were used to genotype positive samples. There was no isolation of the parasite by bioassay in mice. T. gondii was not detected in any of the groups of mussels by nested-PCR. DNA of T. gondii was apparently detected by nested-PCR in 2 groups of oysters (3.3%). Genotyping of these two positive samples was not successful. The results suggest that oysters of the species C. rhizophorae, the most common species from the coast of Sao Paulo, can filter and retain T. gondii oocysts from the marine environment. Ingestion of raw oysters as a potential transmission source of T. gondii to humans and marine mammals should be further investigated. (C) 2010 Elsevier B.V. All rights reserved.

Al(2)O(3)/GdAlO(3)fiber for dental porcelain reinforcement

The aim of this study was to test the hypothesis that the addition of continuous or milled GdAlO(3)/Al(2)O(3) fibers to a dental porcelain increases its mechanical properties. Porcelain bars without reinforcement (control) were compared to those reinforced with long fibers (30 vol%). Also, disk specimens reinforced with milled fibers were produced by adding 0 (control), 5 or 10 vol% of particles. The reinforcement with continuous fibers resulted in significant increase in the uniaxial flexural strength from 91.5 to 217.4 MPa. The addition of varied amounts of milled fibers to the porcelain did not significantly affect its biaxial flexural strength compared to the control group. SEM analysis showed that the interface between the continuous fiber and the porcelain was free of defects. On the other hand, it was possible to note the presence of cracks surrounding the milled fiber/porcelain interface. In conclusion, the reinforcement of the porcelain with continuous fibers resulted in an efficient mechanism to increase its mechanical properties; however the addition of milled fibers had no significant effect on the material because the porcelain was not able to wet the ceramic particles during the firing cycle. (C) 2008 Elsevier Ltd. All rights reserved.

Kinetics of fluoride removal from plasma and bone of rats after chronic intake of fluoride

This study evaluated the kinetics of fluoride in plasma, femur surface and the whole femur of rats, after chronic exposure to different water fluoride levels was interrupted. Four groups of Wistar rats received drinking water containing 0, 5, 15 or 50 mu g F/ml for 60 days (n = 50/group). The animals were euthanized immediately after exposure to fluoride or after 7, 30, 90 or 180 days (n = 10/subgroup). Plasma and femurs were collected. Fluoride on the femur surface, whole femur and plasma was analyzed with an electrode. Data were analyzed using ANOVA and Tukey`s test (p < 0.05). The increase in plasma fluoride levels was significant only for the 50 mu g F/ml group at 0 and 7 days. Regarding bone surface and whole bone, for most groups, significant increases in fluoride concentrations were observed with the increase in water fluoride concentrations at each time of euthanasia. For fluoride doses up to 15 mu g F/ml, femur surface fluoride levels were reestablished 180 days after the exposure was discontinued, which Was not valid for whole femur or for higher fluoride doses. We found a different kinetics of fluoride in plasma,femur surface and the whole femur of rats after chronic exposure to fluoride is interrupted. Copyright 2008 Prous Science, S.A.U. or its licensors. All rights reserved.

Dental caries in adolescents from public schools in Maputo, Mozambique

Aim: To assess dental caries prevalence in adolescents at urban and sub-urban areas of Maputo-City, Mozambique and to identify its relationship with dental fluorosis, dental plaque, nutritional status, frequency of sugar consumption and the concentration of fluoride in public water supply. Methods: Subjects (n=601) were randomly selected from five urban schools and five sub-urban schools. Clinical examinations were performed under standardised conditions by a trained examiner using DMFT index, SiC index, fluorosis index, PHP, BMI, a sugar consumption questionnaire and water supply analysis. The bivariate analysis and Pearson correlation was used (p<0.05). Results: The mean (DMFT) was 0.9 (+/- 1.65 SD). Children in urban schools showed less dental caries (0.8 +/- 1.49SD) than children in sub-urban schools (1.1 +/- 1.80SD, p=0.03). Only 8.15% had very mild to moderate fluorosis but most presented poor oral hygiene. Cases of malnutrition were found in more sub-urban schools (n = 109; 36.22%) than in urban schools (n = 66; 22.00%) (p=0.03). The frequency of sugar consumption was higher among urban children compared to suburban schools (p<0.00). The level of fluoride in water consumption in urban schools was 0.4 ppmF, above the level of fluoride in sub-urban schools, 0.2 ppmF. Conclusion: Dental caries should not be considered a major oral health problem in Maputo at the moment. However the data suggest the implementation of a population strategy to reduce dental caries rates, in children of both urban and sub-urban areas, in Maputo.

Effects of light exposure time on composite resin hardness after root reinforcement using translucent fibre post

Objectives: The purpose of this in vitro study was to evaluate the Vickers hardness (VHN) of a Light Core (Bisco) composite resin after root reinforcement, according to the light exposure time, region of intracanal reinforcement and lateral distance from the light-transmitting fibre post. Methods: Forty-five 17-mm long roots were used. Twenty-four hours after obturation, the root canals were emptied to a depth of 12 mm and the root dentine was artificially flared to produce a 1 mm space between the fibre post and the canal walls. The roots were bulk restored with the composite resin, which was photoactivated through the post for 40 s (G1, control), 80 s (G2) or 120 s (G3). Twenty-four hours after post-cementation, the specimens were sectioned transversely into three slices at depths of 2, 6 and 10 mm, corresponding to the coronal, middle and apical regions of the reinforced root. Composite VHN was measured as the average of three indentations (100 g/15 s) in each region at lateral distances of 50, 200 and 350 mu m from the cement/post-interface. Results: Three-way analysis of variance (alpha = 0.05) indicated that the factors time, region and distance influenced the hardness and that the interaction time x region was statistically significant (p = 0.0193). Tukey`s test showed that the mean VHN values for G1 (76.37 +/- 8.58) and G2 (74.89 +/- 6.28) differed significantly from that for G3 (79.5 +/- 5.18). Conclusions: Composite resin hardness was significantly lower in deeper regions of root reinforcement and in lateral areas distant from the post. Overall, a light exposure time of 120 s provided higher composite hardness than the shorter times (40 and 80 s). (C) 2008 Elsevier Ltd. All rights reserved.

Dynamics of Aeromonas species isolated from wastewater treatment system

Aeromonas are widely distributed in the aquatic environment, and are considered to be emerging organisms that can produce a series of virulence factors. The present study was carried out in a sanitary sewage stabilization pond treatment system, located in Lins, State of Sao Paulo, Brazil. Most probable number was applied for estimation of the genus Aeromonas. Colony isolation was carried out on blood agar ampicillin and confirmed by biochemical characterization. Aeromonas species were isolated in 72.4% of influent samples, and in 55.2 and 48.3% of effluent from anaerobic and facultative lagoons, respectively. Thirteen Aeromonas species were isolated, representing most of the recognized species of these organisms. Even though it was possible to observe a tendency of decrease, total elimination of these organisms from the studied system was not achieved. Understanding of the pathogenic organism`s dynamics in wastewater treatment systems with a reuse potential is especially important because of the risk it represents.

Increasing consumption of ultra-processed foods and likely impact on human health: evidence from Brazil

Objective: To assess time trends in the contribution of processed foods to food purchases made by Brazilian households and to explore the potential impact on the overall quality of the diet. Design: Application of a new classification of foodstuffs based on extent and purpose of food processing to data collected by comparable probabilistic household budget surveys. The classification assigns foodstuffs to the following groups: unprocessed/minimally processed foods (Group 1); processed culinary ingredients (Group 2); or ultra-processed ready-to-eat or ready-to-heat food products (Group 3). Setting: Eleven metropolitan areas of Brazil. Subjects: Households; n 13 611 in 1987-8, n 16 014 in 1995-5 and n 13 848 in 2002-3. Results: Over the last three decades, the household consumption of Group 1 and Group 2 foods has been steadily replaced by consumption of Group 3 ultra-processed food products, both overall and in lower- and upper-income groups. In the 2002-3 survey, Group 3 items represented more than one-quarter of total energy (more than one-third for higher-income households). The overall nutrient profile of Group 3 items, compared with that of Group 1 and Group 2 items, revealed more added sugar, more saturated fat, more sodium, less fibre and much higher energy density. Conclusions: The high energy density and the unfavourable nutrition profiling of Group 3 food products, and also their potential harmful effects on eating and drinking behaviours, indicate that governments and health authorities should use all possible methods, including legislation and statutory regulation, to halt and reverse the replacement of minimally processed foods and processed culinary ingredients by ultra-processed food products.

Surface properties of eucalyptus pulp fibres as reinforcement of cement-based composites

The objective of the present work is to evaluate the effects of the surface properties of unrefined eucalyptus pulp fibres concerning their performance in cement-based composites. The influence of the fibre surface on the microstructure of fibre-cement composites was evaluated after accelerated ageing cycles, which simulate natural weathering. The surface of unbleached pulp is a thin layer that is rich in cellulose, lignin, hemicelluloses, and extractives. Such a layer acts as a physical and chemical barrier to the penetration of low molecular components of cement. The unbleached fibres are less hydrophilic than the bleached ones. Bleaching removes the amorphous lignin and extractives from the surface and renders it more permeable to liquids. Atomic force microscopy (AFM) helps in understanding the fibre-cement interface. Bleaching improved the fibre- cement interfacial bonding, whereas fibres in the unbleached pulp were less susceptible to the re-precipitation of cement hydration products into the fibre cavities (lumens). Therefore, unbleached fibres can improve the long-term performance of the fibre-cement composite owing to their delayed mineralization.

Risk of Giardia infection for drinking water and bathing in a peri-urban area in Sao Paulo, Brazil

A high incidence of waterborne diseases is observed worldwide and in order to address contamination problems prior to an outbreak, quantitative microbial risk assessment is a useful tool for estimating the risk of infection. The objective of this paper was to assess the probability of Giardia infection from consuming water from shallow wells in a peri-urban area. Giardia has been described as an important waterborne pathogen and reported in several water sources, including ground waters. Sixteen water samples were collected and examined according to the US EPA (1623, 2005). A Monte Carlo method was used to address the potential risk as described by the exponential dose response model. Giardia cysts occurred in 62.5% of the samples (0.1-36.1 cysts/l). A median risk of 10-1 for the population was estimated and the adult ingestion was the highest risk driver. This study illustrates the vulnerability of shallow well water supply systems in peri-urban areas.

Electrocombustion of humic acid and removal of algae from aqueous solutions

This paper reports experiments involving the electrochemical combustion of humic acid (HA) and removal of algae from pond water. An electrochemical flow reactor with a boron-doped diamond film anode was used and constant current experiments were conducted in batch recirculation mode. The mass transfer characteristics of the electrochemical device were determined by voltammetric experiments in the potential region of water stability, followed by a controlled current experiment in the potential region of oxygen evolution. The average mass transfer coefficient was 5.2 x 10(-5) m s(-1). The pond water was then processed to remove HA and algae in the conditions in which the reaction combustion occurred under mass transfer control. To this end, the mass transfer coefficient was used to estimate the initial limiting current density applied in the electrolytic experiments. As expected, all the parameters analyzed here-solution absorbance at 270 nm, total phenol concentration and total organic carbon concentration-decayed according to first-order kinetics. Since the diamond film anode successfully incinerated organic matter, the electrochemical system proved to be predictable and programmable.

Biobased composites from tannin-phenolic polymers reinforced with coir fibers

Tannin-phenolic polymers prepared using tannin, a macromolecule obtained from natural sources, were used in the preparation of composites reinforced with coir fibers. The composites based on tannin-phenolic polymers (50% (w/w) of tannin as substitute of the phenol) were prepared using the coir fibers as reinforcement (30-70% (w/w), 3.0-6.0 cm, randomly distributed). The Izod impact strength of the composites showed an improvement in this property due to the incorporation of coir fibers in the tannin-phenolic matrices. The SEM images showed excellent adhesion at the fiber/matrix interface. The coir fiber had bundles regularly spaced, which enhanced the diffusion of the resin into the fiber. In addition, the high lignin content of this fiber results in a high concentration of aromatic rings, which increased the compatibility with the matrix. The values of the diffusion coefficient of water, determined using Fick`s laws, show that there was no correlation between the fiber percentage and the water diffusion. The DMTA curves showed that the storage moduli of the composites reinforced with coir fibers were considerably higher than that of the thermoset, and the increase in the proportion of fibers led to a proportional increase in the storage moduli of these materials. The biobased composites obtained have potential for non-structural applications, such as in the internal parts of automotives vehicles. To our knowledge, this is the first study on this kind of biobased composites. (C) 2010 Elsevier B.V. All rights reserved.