Cytogenetic study in sand spiders (Sicariidae) from the brazilian Caatinga: sex chromosome system diversity in closely related species / Estudio citogenético en arañas de arena (Sicariidae) de la Caatinga brasileña: diversidad del sistema de cromosomas sexuales en especies estrechamente relacionadas

ABSTRACT In this study, we investigated the chromosomes of three species of Sicarius spiders from the Brazilian Caatinga, using classical and molecular cytogenetic techniques. Based on the phylogenetic approach, we also discussed about the variation of diploid number, types of sex chromosome system and changes in the localization of ribosomal genes of Scytodoidea. Sicarius are Synspermiata spiders that together with the genera Loxosceles and Hexophthalma constitute the family Sicariidae. In this group, the available cytogenetic data showed a low diploid number range (2n♂=18 to 2n♂=23) and the presence of only multiple sex chromosome systems (X1X2Y and X1X20). Mitotic metaphase cells exhibited 2n♂=16+X1X2Y for Sicarius cariri and S. ornatus, and 2n♂=18+XY for S. tropicus. In these species, silver impregnation revealed nucleolar organizer region (Ag-NOR) on the terminal region of pair 1. In S. ornatus and S. tropicus, the results obtained with fluorescent in situ hybridization (FISH) using 18S rDNA probe were similar to Ag-NOR, however in S. cariri, the ribosomal sites were localized in the terminal region of the X1 sex chromosome. In this work, we presented the first description of a simple sex chromosome system for Sicariidae, helping to understand how the XY sex chromosome system evolved from the X1X2Y system. Additionally, FISH data incongruous with Ag-NOR indicate that the cytogenetic studies in Sicariidae allow investigating the relation between the karyotype evolution and the distribution and the activity of rDNA genes.

RESUMEN En este estudio, investigamos los cromosomas de tres especies de arañas Sicarius de la Caatinga brasileña, utilizando técnicas de citogenética clásica y molecular. Usando un enfoque filogenético, también discutimos la variación del número diploide, los tipos de sistema cromosómico sexual y los cambios en la localización de los genes ribosómicos en Scytodoidea. Los Sicarius son arañas Synspermiata que, junto con los géneros Loxosceles y Hexophthalma, constituyen a la familia Sicariidae. En este grupo, los datos citogenéticos disponibles mostraron un rango de número diploide bajo (2n♂=18 a 2n♂=23) y únicamente la presencia de sistemas de cromosomas sexuales múltiples (X1X2Y y X1X20). Las células mitóticas en metafase mostraron 2n♂=16+X1X2Y para Sicarius cariri y S. ornatus, y 2n♂=18+XY para S. tropicus. En estas especies, la impregnación de plata reveló la región organizadora nucleolar (Ag-NOR) en la región terminal del par 1. En S. ornatus y S. tropicus, los resultados obtenidos con la hibridación in situ fluorescente (FISH) utilizando la sonda de ADNr 18S fueron similares a los de Ag-NOR, sin embargo, en S. cariri los sitios ribosomales se localizaron en la región terminal del cromosoma sexual X1. En este trabajo, presentamos la primera descripción de un sistema cromosómico sexual simple para Sicariidae, ayudando a entender cómo el sistema cromosómico sexual XY evolucionó a partir del sistema X1X2Y. Además, los datos de FISH incongruentes con Ag-NOR indican que los estudios citogenéticos en Sicariidae permiten investigar la relación entre la evolución del cariotipo y la distribución y la actividad de los genes de ADNr.

Inflammatory profile of apical periodontitis associated with liver fibrosis in rats: histological and immunohistochemical analysis.

AIM: To investigate the effects of liver fibrosis (LF) on the pro-inflammatory mediators and periapical bone resorption of apical periodontitis (AP) in rats. METHODOLOGY: Forty male Wistar rats were distributed into four groups: C - control, AP - rats with AP, LF - rats with LF, AP + LF - rats with AP and LF. LF was induced by carbon tetrachloride administration for 8 weeks and surgical bile duct ligation for 4 weeks; AP was induced in the teeth of rats by dental pulp exposure to the oral environment for 30 days. Jaws and livers were removed after euthanasia. Haematoxylin and Eosin (H&E) and Picrosirius Red (PSR) staining were used to confirm fibrosis in the livers. The jaws were analysed using H&E staining, immunohistochemical assays of interleukin (IL)-1ß, IL-6 and tumour necrosis factor-alpha (TNF-α). Student's t-test and Mann-Whitney's U-test were used for statistical analysis (P < 0.05). RESULTS: Inflammatory infiltrate was moderate in the AP group and severe in the AP + LF group (P < 0.05). Periapical bone resorption was significantly larger in the AP + LF group compared with the AP group (P < 0.05). IL-1ß, IL-6 and TNF-α levels were significantly higher in AP + LF group when compared to the AP group (P < 0.05). CONCLUSION: More intense inflammatory infiltrate, greater amounts of pro-inflammatory cytokines and increased periapical bone resorption were observed in the presence of liver fibrosis in rats with exposed pulps.

Predictors of nutritional and inflammation risk in hemodialysis patients.

BACKGROUND: Malnutrition and chronic inflammation are prevalent complications in hemodialysis (HD) patients. Different nutritional assessment tools are used to identify patients at risk. A composite and comprehensive malnutrition inflammation score (MIS) has been correlated with morbidity and mortality, and appears to be a robust and quantitative tool. OBJECTIVES: Determine malnutrition risk profile in a sample of portuguese HD patients; determine the association of clinical and laboratory factors with MIS, and the impact of each parameter on MIS. METHODS AND RESULTS: We performed, between September 15th of 2015 and January 31st of 2016, a cross sectional analysis of 2975 patients, representing 25% of portuguese HD patients. 59% were men (66.7 ± 14.8 years); 31% diabetic; 79% and 21% performed, respectively, high-flux HD and HDF. A MIS >5 was considered to indicate higher risk and was present in 1489 patients (50%). Amongst all parameters, comorbilities/dialysis vintage, transferrin, functional capacity, changes in body weight and decreased fat stores showed the higher impact, while albumin had one of the lowest impact on the nutritional risk. MULTIVARIABLE ANALYSIS: Higher age (>75 years, OR 1.71, p < 0.001), diabetes (OR 1.25, p = 0.026), lower P levels (OR 1.57,p = 0.001), higher Ca levels (OR 1.51, p < 0.001), higher ERI (OR 1.05, p < 0.001), higher Kt/V (OR 2.14, p < 0.001) and higher CRP (OR 1.01, p < 0.001) were independently associated with a higher risk of MIS>5; higher nPNA (OR 0.29, p < 0.001) and higher Pcreat (OR 0.88, p < 0.001) were associated with a risk reduction of MIS>5 (95% CI). CONCLUSIONS: Routine clinical and analytic parameters were found to be associated with MIS range that might indicate higher risk, and may represent a simple alert sign for the need of further assessments.

Palatogingival groove and root canal instrumentation.

AIM: To evaluate the morphology and impact of root canal preparation in maxillary incisors with palatogingival grooves (PGG). METHODOLOGY: Twenty extracted human maxillary incisors with PGG were subjected to macroscopic analysis and scanning electron microscopy analysis (SEM). The following characteristics of the PGG were evaluated: depth, point of origin in the cingulum, extension and position on the lingual surface. Furthermore, the presence of calculus, communications between the root canal system and the PGG, and root resorptions were investigated. The root canals were subsequently instrumented with K-files of three consecutive sizes. The teeth were sectioned, and the axial plane of each tooth section was imaged using SEM before and after instrumentation. The distance between the root canal walls and the PGG was calculated according to the location. Additionally, the distance between canal walls and cementum was measured at three different sites, to verify if instrumentation influenced dentine removal on a specific wall in teeth with PGG. Statistical analysis was performed using the Mann-Whitney or Student's t-test (P < 0.05). RESULTS: Macroscopic analysis revealed that a deep groove was most frequently observed (75%), followed by a depression/shallow groove (25%) (P < 0.05). PGG typically originated in the distal margin ridge of the cingulum (65%) (P < 0.05), extending only to the middle (45%) or up to the apical (50%) third of the root (P < 0.05). Additionally, PGGs were typically located on the distal aspect of the lingual surface (70%) (P < 0.05). Calculus was concentrated on the surface of the crown and cementum-enamel junction (P < 0.05). Communication between the root canal and PGG was present in 35% of teeth, and root resorptions were noted in 50% of teeth. The distance between the external root surface and the pulp cavity was significantly narrower after instrumentation (P < 0.05); however, root canal preparation did not influence dentine removal on the specific wall associated with the groove (P > 0.05). CONCLUSIONS: Palatogingival grooves were characteristically deep and originated from the distal margin of the cingulum. Although it has been associated with a thinner root wall, root canal preparation did not influence the thickness of the specific wall in the maxillary incisors with PGG.

3D map distribution of metallic nanoparticles in whole cells using MeV ion microscopy.

In this work, a new tool was developed, the MORIA program that readily translates Rutherford backscattering spectrometry (RBS) output data into visual information, creating a display of the distribution of elements in a true three-dimensional (3D) environment. The program methodology is illustrated with the analysis of yeast Saccharomyces cerevisiae cells, exposed to copper oxide nanoparticles (CuO-NP) and HeLa cells in the presence of gold nanoparticles (Au-NP), using different beam species, energies and nuclear microscopy systems. Results demonstrate that for both cell types, the NP internalization can be clearly perceived. The 3D models of the distribution of CuO-NP in S. cerevisiae cells indicate the nonuniform distribution of NP in the cellular environment and a relevant confinement of CuO-NP to the cell wall. This suggests the impenetrability of certain cellular organelles or compartments for NP. By contrast, using a high-resolution ion beam system, discretized agglomerates of Au-NP were visualized inside the HeLa cell. This is consistent with the mechanism of entry of these NPs in the cellular space by endocytosis enclosed in endosomal vesicles. This approach shows RBS to be a powerful imaging technique assigning to nuclear microscopy unparalleled potential to assess nanoparticle distribution inside the cellular volume.

Hypertensive disorders during pregnancy and health outcomes in the offspring: a systematic review.

UNLABELLED: The hypertensive disorders of pregnancy complicate up to 10% of pregnancies worldwide and are a leading cause of maternal, foetal, and neonatal morbidity and mortality. The aim of this study was to present an overview of recent studies addressing offspring's medium and long-term health outcomes after intrauterine exposure to maternal hypertension. A search on PubMed/MEDLINE and Bireme databases was conducted to identify observational studies that reported any offspring outcome measured after the 6th month of life. The search was limited to studies published after May 2008. Forty-five articles were included and categorized into four groups of outcomes: cardiovascular, immune, metabolic and behavioural/neurological effects. According to our findings, hypertensive disorders of pregnancy had an overall negative impact on offspring's cardiovascular, immune and neurological health, although not all parameters analysed in each group had consistent results among studies. The most prominent and reliable associations were verified between gestational hypertension and higher offspring's blood pressure and between preeclampsia and offspring's lower cognitive functioning. In the metabolic outcomes, body composition had conflicting results among papers, while all studies that examined blood biomarkers showed no evidence that preeclampsia or gestational hypertension could be associated with an alteration of this metabolic outcomes. Most included studies were highly heterogeneous regarding the measure of outcomes and covariables used for adjustments. Future studies should consider using the same protocols and cut-off points already published so that results can be better compared and summarized. This review was registered in PROSPERO. REGISTRATION NUMBER: CRD42015020838.

Selection of reference genes for expression analyses of red-fleshed sweet orange (Citrus sinensis).

Red-fleshed oranges (Citrus sinensis) contain high levels of carotenoids and lycopene. The growing consumer demand for products with health benefits has increased interest in these types of Citrus cultivars as a potential source of nutraceuticals. However, little is known about the physiology of these cultivars under Brazilian conditions. Transcriptome and gene expression analyses are important tools in the breeding and management of red-fleshed sweet orange cultivars. Reverse transcription quantitative polymerase chain reaction is a method of quantifying gene expression, but various standardizations are required to obtain precise, accurate, and specific results. Among the standardizations required, the choice of suitable stable reference genes is fundamental. The objective of this study was to evaluate the stability of 11 candidate genes using various tissue and organ samples from healthy plants or leaves from citrus greening disease (Huanglongbing)-symptomatic plants of a Brazilian red-fleshed cultivar ('Sanguínea de Mombuca'), in order to select the most suitable reference gene for investigating gene expression under these conditions. geNorm and NormFinder identified genes that encoded translation initiation factor 3, ribosomal protein L35, and translation initiation factor 5A as the most stable genes under the biological conditions tested, and genes coding actin (ACT) and the subunit of the PSI reaction center subunit III were the least stable. Phosphatase, malate dehydrogenase, and ACT were the most stable genes in the leaf samples of infected plants.

Effects of sodium hypochlorite associated with EDTA and etidronate on apical root transportation.

AIM: To evaluate the influence of sodium hypochlorite associated with EDTA and etidronate on apical root transportation. METHODOLOGY: Forty-five roots of human mandibular molars with curvatures of 15-25° were embedded in acrylic resin to allow standardized angulation of the initial and final radiographs. The pre-instrumentation radiographs of the mesiobuccal canal of each root were taken using a radiograph digital sensor with a size 15 K-file in the canal. The canals were prepared with the ProTaper Universal system (Dentsply Maillefer, Ballaigues, Switzerland), using one of the following irrigation regimens during the instrumentation (n = 15): G1 - irrigation with 20 mL of saline solution (control); G2 - alternating irrigation with 2.5% hypochlorite solution (NaOCl) (15 mL); and 17% ethylenediaminetetraacetic acid (EDTA) (5 mL). During instrumentation, the canal was filled with NaOCl and then between each exchange of instrument filled with EDTA for 1 min, and G3 - irrigation with 20 mL of 5% NaOCl and 18% etidronate solution (HEBP) mixed in equal parts. The postinstrumentation radiographs were made with a F3 instrument in the canal. The images were magnified and superposed with Adobe Photoshop software (Adobe Systems, Mountain View, CA, USA). Apical transportation was determined with AutoCAD 2012 software (Autodesk Inc., San Rafael, CA, USA) by measuring the distance in millimetres between the tips of the instruments. The results were subjected to the nonparametric statistical Kruskal-Wallis test (α < 0.05). RESULTS: The median transportation and interquartile range values were 0.00 ± 0.05 for G1, 0.08 ± 0.23 for G2 and 0.13 ± 0.14 for G3. Comparison between groups showed that apical transportation in G3 was significantly greater than in G1 (P < 0.05). CONCLUSION: The use of NaOCl associated with etidronate increased apical transportation in the canals of extracted teeth.

Comparative study of the ovarian morphology in the order Polydesmida (Diplopoda) and description of unusual structures in the female reproductive system / Estudo comparativo da morfologia ovariana na ordem Polydesmida (Diplopoda) e descrição de estruturas incomuns do sistema reprodutor feminino

Polydesmida is the largest order in the class Diplopoda in terms of the number of species, genera and families, but there are few studies of the ovarian morphology of species in this order. This study aimed to perform a comparative study of the ovarian morphology of three species from this order to increase the understanding of the morphological evolution of this system in Polydesmida. Adults females of two of these species, Poratia salvator and Myrmecodesmus hastatus, belonging to the family Pyrgodesmidae, had a unpaired ovary that formed a tubular organ containing oocytes, with P. salvator present grouping of oocytes into a structure similar to ovisacs. This condition appeared to be apomorphic and was associated with the small size of these species ( 10 mm) and their short life cycle. The third species, Telonychopus klossae (Chelodesmidae), had a large body and an ovary with paired ovisacs, which was symplesiomorphic with other Polydesmida families.

A ordem Polydesmida é a maior da classe Diplopoda em número de espécies, gêneros e famílias, entretanto existem poucos estudos sobre a morfologia ovariana de suas espécies. Esta pesquisa tem como objetivo realizar um estudo comparativo sobre a morfologia ovariana de três espécies dessa ordem a fim de contribuir para discussões sobre a evolução morfológica deste sistema no grupo. Duas dessas espécies, Poratia salvator e Myrmecodesmus hastatus, representantes da família Pyrgodesmidae, apresentam ovário como um órgão tubular único, com a primeira possuindo os ovócitos agrupados em uma estrutura semelhante a ovissacos. Esta condição parece ser uma apomorfia aparentemente associada ao tamanho diminuto dos animais destas espécies (menos de 10 mm) e suas histórias de vida curtas. A terceira espécie, Telonychopus klossae, um quelodesmídeo de tamanho corporal grande, possui um ovário pareado com presença de ovissacos, condição simplesiomórfica semelhante à de outras famílias de Polydesmida.

Biomarkers of exposure to metal dust in exhaled breath condensate: methodology optimization.

In occupational assessments where workers are exposed to metal dust, the liquid condensate of exhaled breath (EBC) may provide unique indication of pulmonary exposure. The main goal of this study was to demonstrate the quality of EBC to biological monitoring of human exposure. A pilot study was performed in a group of metal dust-exposed workers and a group of nonexposed individuals working in offices. Only metal dust-exposed workers were followed along the working week to determine the best time of collection. Metal analyses were performed with inductively coupled plasma mass spectrometry (ICP-MS). Analytical methodology was tested using an EBC sample pool for several occupationally exposed metals: potassium, chromium, manganese, copper, zinc, strontium, cadmium, antimony, and lead. Metal contents in EBC of exposed workers were higher than controls at the beginning of the shift and remained augmented throughout the working week. The results obtained support the establishment of EBC as an indicator of pulmonary exposure to metals.

Assessment of exposure to metals in lead processing industries.

Inhalation of particulate matter in industrial environments has been associated with respiratory symptoms and lung diseases, which continues to lead to long- and short-term hazardous health effects on exposed subjects. The main objectives of this study were (a) to determine the dust exposure of workers from the lead industry in different operations and (b) to evaluate if the Exhaled Breath Condensate (EBC) can be used as a non-invasive tool to evaluate this exposure. Therefore, this cross-sectional study not only measured the exposure to Airborne Particulate Matter (APM) and to the associated elements but also analysed the EBC elemental composition. APM was collected in Ind1, Ind2, Offices and outdoor with Gent samplers, which delivers two size fractions: fine particulate matter (< 2.5 µm), and coarse particulate matter (between 2.5 and 10 µm). EBC samples were collected from the workers and from a non-exposed group working in Offices. The techniques INAA and PIXE were used for the APM element characterization and ICP-MS for EBC elemental content. The PM2.5 and PM2.5-10 mass concentrations were significantly higher in the industries studied than in Offices and in the environment. At the industrial sites surveyed the coarse fraction dominated and both factories had different fingerprints: APM elements with higher expression were Pb, Sb, Na, Cl and Fe in Ind1 and Pb, Si, Br, Ca, Al, Cl and Na in Ind2. Most of these elements revealed a gradient of concentration where Ind.1 > Ind.2 > Offices and EBC revealed a clear translation of this exposure, suggesting the latter to be a potential good indicator of exposure to metals in occupational settings. Pb in EBC presented the most representative results. Even though EBC was found to reflect predominantly the inhaled coarser fraction it is more related to concentration levels of exposure than to the predominance of APM fraction. The present study demonstrated not only the ability of EBC to reflect environmental exposure to metals but also the importance of measuring and characterizing different fractions of APM for a correct assessment.

Comparative study of the ovarian morphology in the order Polydesmida (Diplopoda) and description of unusual structures in the female reproductive system.

Polydesmida is the largest order in the class Diplopoda in terms of the number of species, genera and families, but there are few studies of the ovarian morphology of species in this order. This study aimed to perform a comparative study of the ovarian morphology of three species from this order to increase the understanding of the morphological evolution of this system in Polydesmida. Adults females of two of these species, Poratia salvator and Myrmecodesmus hastatus, belonging to the family Pyrgodesmidae, had a unpaired ovary that formed a tubular organ containing oocytes, with P. salvator present grouping of oocytes into a structure similar to ovisacs. This condition appeared to be apomorphic and was associated with the small size of these species (< 10 mm) and their short life cycle. The third species, Telonychopus klossae (Chelodesmidae), had a large body and an ovary with paired ovisacs, which was symplesiomorphic with other Polydesmida families.

Phenotypic and genetic parameters compared during repeated measures of longissimus muscle area and subcutaneous fat thickness in Nelore cattle.

Real-time ultrasound is currently used for in vivo carcass evaluation of beef cattle. We estimated heritability and repeatability coefficients for ultrasound-measured carcass traits in Nelore cattle. We measured longissimus muscle area, backfat thickness, and rump fat thickness in male and female animals ranging in age from 10 to 26 months. The variance components were estimated by single-trait analysis using the derivative-free restricted maximum likelihood method, under an animal model. The complete data file included 3077 records of 1172 animals born between 2003 and 2008. Two analyses were performed using a repeatability model: a) all records of longissimus muscle area, backfat thickness, and rump fat thickness of animals aged 10 to 26 months, and b) considering two age periods (11 to 17 and 18 to 24 months). The heritability estimates ranged from 0.33 to 0.46 for longissimus muscle area, from 0.20 to 0.26 for backfat, and from 0.26 to 0.29 for rump fat, demonstrating that direct selection for these traits can provide genetic gain. The repeatability estimates were moderate, ranging from 0.42 to 0.73. The highest repeatability estimates were obtained for longissimus muscle area in both the first (0.70) and second (0.73) analyses. The repeatability estimates suggest that ultrasound measures are precise; higher precision was obtained for longissimus muscle area than for subcutaneous fat thickness. The latter could be obtained during periods of greater feed availability to increase precision.

Establishing references for gene expression analyses by RT-qPCR in Theobroma cacao tissues.

Lack of continuous progress in Theobroma cacao (Malvaceae) breeding, especially associated with seed quality traits, requires more efficient selection methods based on genomic information. Reverse transcript quantitative PCR (RT-qPCR) has become the method of choice for gene expression analysis, but relative expression analysis requires various reference genes, which must be stable across various biological conditions. We sought suitable reference genes for various tissues of cacao, especially developing seeds. Ten potential reference genes were analyzed for stability at various stages of embryo development, leaves, stems, roots, flowers, and pod epicarp; seven of them were also evaluated in shoot tips treated either with hormones (salicylate; ethefon; methyl-jasmonate) or after inoculation with the fungus Moniliophthora perniciosa (Marasmiaceae sensu lato). For developing embryos, the three most stable genes were actin (ACT), polyubiquitin (PUB), and ribosomal protein L35 (Rpl35). In the analyses of various tissues, the most stable genes were malate dehydrogenase (MDH), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and acyl-carrier protein B (ACP B). GAPDH, MDH and tubulin (TUB) were the most appropriate for normalization when shoot apexes were treated with hormones, while ACT, TUB and Rpl35 were the most appropriate after inoculation with M. perniciosa. We conclude that for each plant system and biological or ontogenetical condition, there is a need to define suitable reference genes. This is the first report to define reference genes for expression studies in cacao.

Fertility tables of two populations of the parthenogenetic species Poratia salvator (Diplopoda, Polydesmida, Pyrgodesmidae).

Diplopods are very susceptible to various degrees of environmental disturbance, particularly climate, altitude and diet. In order to increase our understanding of millipede ecological plasticity, we used fertility tables to access and to compare the fertility and survival of two populations of the parthenogenetic species Poratia salvator from two areas with distinct characteristics. Collecting was conducted in two localities within the state of Mato Grosso, in the Pantanal of Mato Grosso, municipality of Nossa Senhora do Livramento, and in the municipality of Várzea Grande. The specimens were maintained at room temperature. In the first generation, individuals from the Pantanal of Mato Grosso population reproduced early and also died earlier when compared with individuals from the Várzea Grande population. Furthermore, the population from the Pantanal had a lower net reproduction rate and a higher intrinsic growth rate than the population from the Várzea Grande. The generation time was lower for the Pantanal population than for the Várzea Grande population. In the second generation, the net reproduction rate observed for both populations was higher than that observed in the first generation, suggesting an increase in the reproductive potential of the females throughout their lives. The intrinsic growth rate of both populations decreased as a function of an increase in generation time observed in the second generation. As a result, the population growth rate in the second generation was slower when compared with the first generation, probably a result of the longer life expectancy of the second generation, which may have become adapted to the artificial conditions of the experiment.

Microsatellite polymorphisms in cassava landraces from the Cerrado biome, Mato Grosso do sul, Brazil.

Using nine microsatellite loci, we investigated genetic structure and diversity in 83 Brazilian cassava accessions, including several landraces, in the Cerrado biome in Mato Grosso do Sul, Brazil. All nine loci were polymorphic, averaging 6.00 alleles per locus. Treating each of seven municipalities as a cassava group or population, they averaged 3.5 alleles per locus, with 97% polymorphic loci, high values for observed heterozygosity (0.32) and gene diversity (0.56). Total genetic variability was high (0.668), and most of this genetic variability was concentrated within municipalities (0.577). Cluster and structure analyses divided accessions into two major clusters or populations (K = 2). Also, a significant genetic versus geographic correlation was found (r = 0.4567; P < 0.0260). Migratory routes in the Cerrado are considered main contributors to the region's high cassava diversity and spatial genetic structure, amplifying interactions between traditional farmers and the evolutionary dynamics of this crop.

Stratum corneum is an effective barrier to TiO2 and ZnO nanoparticle percutaneous absorption.

BACKGROUND: There is increasing concern over the local and systemic side effects of TiO(2) and ZnO coated nanoparticles widely used in sun blockers. OBJECTIVE: To determine the localization and possible skin penetration of TiO(2) and ZnO nanoparticles, dispersed in 3 sunscreen formulations, under realistic in vivo conditions in normal and altered skin. METHODS: Nuclear microscopy techniques provided spatially resolved quantitative analysis of Ti and Zn nanoparticle distributions in transversal cryosections of skin obtained by biopsy with no further treatment. A test hydrophobic formulation containing coated 20-nm TiO(2) nanoparticles and 2 commercial sunscreen formulations containing TiO(2) alone or in combination with ZnO were tried, taking into account realistic use conditions by consumers and compared with the recommended standard condition for the sun protection factor test. The protocols consisted of an open test. RESULTS: Following a 2-hour exposure period of normal human skin to TiO(2)- and ZnO-containing sunscreens, detectable amounts of these physical blockers were only present at the skin surface and in the uppermost stratum corneum regions. Layers deeper than the stratum corneum were devoid of TiO(2) or exogenous ZnO, even after 48 h of exposure to the sunscreen, under occlusion. Deposition of TiO(2) and ZnO nanoparticles in the openings of the pilosebaceous follicles was also observed, suggesting a preferential fixation area. Penetration of nanoparticles into viable skin tissue could not be detected. CONCLUSIONS: TiO(2) or ZnO nanoparticles are absent or their levels are too low to be tested under the stratum corneum in human viable epidermal layers. Therefore, significant penetration towards the underlying keratinocytes is unlikely.

An assessment of time-dependent effects of lead exposure in algerian mice (Mus spretus) using different methodological approaches.

Time-dependent effects of lead (Pb) toxicity were studied in Algerian mice (Mus spretus) treated with Pb acetate via drinking water (1 g Pb acetate/L) for different periods of exposure (15, 45, and 90 d). End points included the determination of hepatic Pb concentration and the assessment of some morphophysiological, biochemical and cytogenetical parameters. A control group receiving distilled water was also monitored for comparative purposes. Hepatic Pb accumulation increased with the time of exposure and was significantly higher in treated mice when compared to controls. In association with significant body mass loss in Pb-exposed mice, for 15 and 45 d, a significant increase in the relative spleen mass was observed after 45 d of intoxication. Pb-exposed mice also showed significant decreases in red blood cells, hematocrit, and mean corpuscular hemoglobin. On the contrary, changes in plasma transferases (aspartate aminotransferase and alanine aminotransferase) and hepatic superoxide dismutase activities did not reach statistical significance. A significant increase in the frequency of micronucleated polychromatic bone marrow erythrocytes was also found in the 90-d-exposed mice, compared to nontreated mice and the other exposed groups. Exposure to Pb acetate resulted also in a slight time-dependent decrease of the polychromatic-normochromatic ratio. These results support the concept that a long-term chronic exposure to Pb induced alterations upon some morphophysiological and genetic parameters in Algerian mice.

Elemental distributions in femoral bone of rat under osteoporosis preventive treatments.

One of the abnormalities of bone architecture is osteoporosis as occurring in post-menopausal women. Especially long bones, such as femur, become more fragile and more prone to fracture. The efficiency of several osteoporosis preventative treatments based on oestrogen and progestin in bone structure and mineral recovery was studied using ovariectomized Wistar rats as an osteoporosis experimental model. Diagonal cross-sections of the proximal epiphysis of femoral bones were analysed using nuclear microscopy techniques in order to map and determine the concentration profiles of P, Ca, S, Fe and Zn from the epiphysis to diaphysis and across the cortical and trabecular bone structures. In control animals (not ovariectomized), the S and Zn contents significantly characterized differences between cortical and trabecular bone structures, whereas P and Ca showed increased gradients from the epiphyseal region to the diaphysis. After ovariectomy the differences observed were differential according to the type of hormonal supplementation. A significant decrease in P and Ca contents and depletion of minor and trace minerals, such as S, Fe and Zn, were found for both cortical and trabecular bone structures after ovariectomy relative to controls. Bone mineral contents were reversed to control levels by synthetic oestrogen supplementation, and combined oestrogen and progesterone treatment. Recovery was more evident in the femoral epiphysis and neck than in the diaphysis. The use of oestrogen alone did not lead to bone recovery after ovariectomy. Alterations in bone mineral composition observed for animals receiving synthetic oestrogen and combined oestrogen and progesterone supplement might reflect beneficial structural changes in critical regions of long bones, mostly affected in post-menopausal osteoporosis.

Elemental characterization of tissues of Octopus vulgaris along the Portuguese coast.

The concentrations of V, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, and Pb were measured in digestive gland (DG), branchial hearts (BH), gill (G), and muscle (M) of Octopus vulgaris Cuvier, 1797 caught in three fishing areas of the Portuguese coast, Viana do Castelo, Cascais, and Santa Luzia, for 2 years. The elemental concentrations measured for the different tissues were in accordance with values reported in the literature. The digestive gland presented high concentration levels of Fe, Cu, and Zn, while the branchial hearts showed elevated levels of V, Ni, Mo, as well as Fe and Cu. Significant variations in As, V, Cu, Mo, and Pb tissue concentrations were observed for animals originated from different sampling sites. Pb and As determined in the digestive gland and branchial hearts of animals from Cascais and Santa Luzia, can reflect local environmental characteristics. The variability observed in the elemental concentrations may be useful to further assess the species susceptibility to environmental conditions.