Bisphenol A alters transcript levels of biomarker genes for major depressive disorder in vascular endothelial cells and colon cancer cells

Bisphenol A (BPA) is capable of mimicking endogenous hormones with potential consequences for human health and BPA exposure has been associated with several human diseases including neuropsychiatric disorders. Here, quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) results show that BPA at low concentrations (10 ng/mL and 1 μg/mL) induces differential transcript levels of four biomarker genes for Major Depressive Disorder (MDD) in HT29 human colon adenocarcinona cell line and Human Umbilical Vein Endothelial Cells (HUVEC). These results substantiate increasing concerns of BPA exposure in levels currently detected in humans.

HSP70 abundance and antioxidant capacity in feeding and fasting gray seal pups: suckling is associated with higher levels of key cellular defenses

Heat shock proteins (HSPs) and antioxidants are key cellular defenses against stress. Seals routinely undergo protracted fasting, which is normally associated with physiological stress in other animals. We tested the hypotheses that (1) relative HSP70 protein abundance is higher in liver and blubber of fasting relative to suckling wild gray seal pups; (2) differences in HSP70 are mirrored in tissue superoxide dismutase (SOD) and catalase activity, as well as glutathione levels; (3) extracellular HSP70 correlates with hepatic and blubber HSP70 abundance; and (4) protein carbonylation, an index of oxidative damage, is lower in tissues with higher levels of these cellular stress markers. In contrast to our expectation, suckling pups had higher relative HSP70 abundance and glutathione levels in liver and blubber and higher hepatic catalase activity. Plasma HSP70 did not correlate with liver or blubber abundance of the protein. Suckling pups did not experience greater protein carbonylation, suggesting that cellular protective mechanisms prevent protein damage despite an apparent increase in cellular stress. SOD activity was not affected by nutritional state, but in blubber tissue, it was positively correlated with blubber thickness. Greater requirements for antioxidants and HSPs in suckling pups or in animals with thicker blubber could arise from rapid protein synthesis, high metabolic fuel availability, and/or exposure to lipophilic toxins. Developmental and nutritional changes in cellular defenses have important implications for gray seals’ susceptibility to additional stress exposure.

Relation between DNA damage measured by comet assay and OGG1 Ser326Cys polymorphism in antineoplastic drugs biomonitoring

Antineoplastic drugs are hazardous chemical agents used mostly in the treatment of patients with cancer, however health professionals that handle and administer these drugs can become exposed and develop DNA damage. Comet assay is a standard method for assessing DNA damage in human biomonitoring and, combined with formamidopyrimidine DNA glycosylase (FPG) enzyme, it specifically detects DNA oxidative damage. The aim of this study was to investigate genotoxic effects in workers occupationally exposed to cytostatics (n = 46), as compared to a control group with no exposure (n = 46) at two Portuguese hospitals, by means of the alkaline comet assay. The potential of the OGG1 Ser326Cys polymorphism as a susceptibility biomarker was also investigated. Exposure was evaluated by investigating the contamination of surfaces and genotoxic assessment was done by alkaline comet assay in peripheral blood lymphocytes. OGG1 Ser326Cys (rs1052133) polymorphism was studied by Real Time PCR. As for exposure assessment, there were 121 (37%) positive samples out of a total of 327 samples analysed from both hospitals. No statistically significant differences (Mann-Whitney test, p > 0.05) were found between subjects with and without exposure, regarding DNA damage and oxidative DNA damage, nevertheless the exposed group exhibited higher values. Moreover, there was no consistent trend regarding the variation of both biomarkers as assessed by comet assay with OGG1 polymorphism. Our study was not statistically significant regarding occupational exposure to antineoplastic drugs and genetic damage assessed by comet assay. However, health professionals should be monitored for risk behaviour, in order to ensure that safety measures are applied and protection devices are used correctly.

Spatial and temporal interaction between sediment and microphytobenthos in a temperate estuarine macro-intertidal bay

Intertidal flats of the estuarine macro-intertidal Baie des Veys (France) were investigated to identify spatial features of sediment and microphytobenthos (MPB) in April 2003. Gradients occurred within the domain, and patches were identified close to vegetated areas or within the oyster-farming areas where calm physical conditions and biodeposition altered the sediment and MPB landscapes. Spatial patterns of chl a content were explained primarily by the influence of sediment features, while bed elevation and compaction brought only minor insights into MPB distribution regulation. The smaller size of MPB patches compared to silt patches revealed the interplay between physical structure defining the sediment landscape, the biotic patches that they contain, and that median grain-size is the most important parameter in explaining the spatial pattern of MPB. Small-scale temporal dynamics of sediment chl a content and grain-size distribution were surveyed in parallel during 2 periods of 14 d to detect tidal and seasonal variations. Our results showed a weak relationship between mud fraction and MPB biomass in March, and this relationship fully disappeared in July. Tidal exposure was the most important parameter in explaining the summer temporal dynamics of MPB. This study reveals the general importance of bed elevation and tidal exposure in muddy habitats and that silt content was a prime governing physical factor in winter. Biostabilisation processes seemed to behave only as secondary factors that could only amplify the initial silt accumulation in summer rather than primary factors explaining spatial or long-term trends of sediment changes.

High diversity of skin-associated bacterial communities of marine fishes is promoted by their high variability among body parts, individuals and species

Animal-associated microbiotas form complex communities, which are suspected to play crucial functions for their host fitness. However, the biodiversity of these communities, including their differences between host species and individuals, has been scarcely studied, especially in case of skin-associated communities. In addition, the intraindividual variability (i.e. between body parts) has never been assessed to date. The objective of this study was to characterize skin bacterial communities of two teleostean fish species, namely the European seabass (Dicentrarchus labrax) and gilthead seabream (Sparus aurata), using a high-throughput DNA sequencing method. In order to focus on intrinsic factors of host-associated bacterial community variability, individuals of the two species were raised in controlled conditions. Bacterial diversity was assessed using a set of four complementary indices, describing the taxonomic and phylogenetic facets of biodiversity and their respective composition (based on presence/absence data) and structure (based on species relative abundances) components. Variability of bacterial diversity was quantified at the interspecific, interindividual and intraindividual scales. We demonstrated that fish surfaces host highly diverse bacterial communities, whose composition was very different from that of surrounding bacterioplankton. This high total biodiversity of skin-associated communities was supported by the important variability, between host species, individuals and the different body parts (dorsal, anal, pectoral and caudal fins).

Maltreatment and Juvenile Delinquency Among Crossover Youth

Objectif. Les jeunes crossover sont définis comme des adolescents qui sont à la fois victimes de maltraitance et délinquants juvéniles. L'objectif de ce mémoire est d’examiner les relations entre les paramètres de la maltraitance (ex., récurrence, sous-types et variété) et les paramètres de la délinquance juvénile (ex., précocité, volume, variété, gravité moyenne et présence de sous-types) vécus par les jeunes crossover. Méthodes. La source des données est constituée de rapports officiels pour tous les adolescents et adolescentes québécois qui ont plaidé coupables ou ont été reconnus coupables d'un crime entre le 1e janvier 2005 et le 31 décembre 2010. D'abord, un portrait des jeunes crossover québécois est dressé à l’aide de statistiques descriptives. Ensuite, des analyses multivariées sont utilisées pour déterminer si les paramètres de la maltraitance prédisent les différentes dimensions de la délinquance et pour examiner les différences selon le sexe. Résultats. La délinquance des jeunes crossover est plus importante que celle des délinquants non-maltraités. Les expériences différentielles de la maltraitance sont liées à des paramètres hétérogènes de la délinquance juvénile. La récurrence de la maltraitance est un prédicteur important des paramètres de la délinquance ultérieure. De plus, la maltraitance est particulièrement influente sur la délinquance des garçons. Implications. Les interventions au sein des systèmes de la protection de la jeunesse et de la justice juvénile doivent être adaptées afin d'identifier les jeunes à risque de délinquance grave, de cibler les dimensions spécifiques de la maltraitance et d’entraver leurs liens à la délinquance ultérieure. L'intervention doit être privilégiée pour les victimes de multiples incidents de maltraitance et pour les garçons victimes de maltraitance.

Relationships between tissue levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), mRNAs and toxicity in the developing male Wistar(Han) rat

We compared the effects of a single acute dose, or chronic fetal exposure, to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the male reproductive system of the Wistar(Han) rat. Tissue samples were taken from dams on GD16 and GD21, and from offspring on PND70 and 120. Steady state concentration of TCDD was demonstrated in the chronic study: body burdens were comparable in both studies. Fetal TCDD concentrations were comparable after acute and chronic exposure, and demonstrate more potent toxicity after chronic versus acute dosing. In maternal liver, cytochrome P450 (CYP)1A1 and CYP1A2 RNA were induced. In fetus, there was induction of both CYP1A1 and CYP1A2 RNA at medium and high doses, but inadequate evidence for induction at low dose in either study. The low level induction of CYP1A1 RNA at low dose in fetus argues against AhR activation in fetus as a mechanism of toxicity of TCDD in causing delay in balanopreputial separation, and the greater induction of CYP1A1 RNA in PND70 offspring liver suggests that lactational transfer of TCDD is crucial to this toxicity. These data characterise the maternal and fetal disposition of TCDD, induction of CYP1A1 RNA as a measure of AhR activation, and suggest that lactational transfer of TCDD determines the difference in delay in balanopreputial separation between the two studies.

Incidence of Chronic and Other Knee Pain in Relation to Occupational Risk Factors in a Large Working Population.

International audience

Identificação e expressão de genes relacionados à via esteroidogênica e resposta antioxidante após a exposição à Atrazina em Poecilia vivipara (Poeciliidae, Cyprinodontiformes)

O herbicida Atrazina (ATR) é um agrotóxico utilizado há cerca de 50 anos, responsável pelo controle seletivo de plantas daninhas em cultivo de arroz, milho e cana-de-açúcar, principalmente. Estudos recentes apontam diversos efeitos desse herbicida em invertebrados e vertebrados, através da contaminação do solo, bem como da lixiviação para os ecossistemas aquáticos. Foi demonstrado que a ATR é um desregulador endócrino, além de causar efeitos como estresse oxidativo, imunotoxicidade e distúrbios no metabolismo energético. No presente estudo, a espécie nativa Poecilia vivipara foi utilizada como modelo experimental para identificar e analisar a expressão de genes atuantes na via esteroidogênica (StAR e Cyp19a1) e genes atuantes no sistema de defesa antioxidante enzimático (SOD-1 e CAT), frente a exposição à diferentes concentrações de ATR. Sequências parciais dos genes-alvo foram obtidas e comparadas com sequências disponíveis de espécies próximas. Foram analisadas a expressão órgãoespecífica para cada um dos genes isolados, bem como a expressão dos genes frente à exposição ao herbicida atrazina. Os animais foram expostos a ATR em concentrações de 2, 10 e 100 µg/L e a expressão dos genes em gônadas e fígado desses animais foram analisadas em 24 e 96 horas de exposição. As sequências obtidas dos genes StAR, Cyp19a1, SOD-1 e CAT apresentaram 821, 80, 954, 350 pares de bases respectivamente, com identidades que variam de 86 a 100% com espécies filogeneticamente próximas a P. vivipara. Os animais apresentaram uma maior expressão dos genes StAR e Cyp19a1 nas gônadas e no fígado, enquanto a menor expressão se mostrou em órgãos como intestino e baço. Já os genes SOD e CAT apresentaram uma maior expressão no fígado, e menor expressão no intestino. Em relação à expressão gênica frente à exposição à ATR, os resultados apontaram para uma indução dos genes StAR, SOD e CAT em 24 horas, nas gônadas e no fígado, enquanto 8 que a expressão do gene Cyp19a1 foi aumentada apenas após 96 horas de exposição. Foi demonstrado que o herbicida ATR, mesmo em baixas concentrações, é capaz de desregular a expressão de genes que codificam tanto para proteínas componentes da via de síntese de hormônios esteróides, quanto para enzimas atuantes na resposta antioxidante celular de P. vivipara. 

Wellbeing and occupational risk perception among health care workers: a multicenter study in Morocco and France

International audience

Karyotype and genome size in Euterpe Mart. (Arecaceae) species.

Euterpe (Martius, 1823), a genus from Central and South America, has species with high economic importance in Brazil, because of their palm heart and fruits, known as açaí berries. Breeding programs have been conducted to increase yield and establish cultivation systems to replace the extraction of wild material. These programs need basic information about the genome of these species to better explore the available genetic variability. The aim of this study was to compare E. edulis (Martius, 1824), E. oleracea (Martius, 1824) and E. precatoria (Martius, 1842), with regard to karyotype, type of interphase nucleus and nuclear DNA amount. Metaphase chromosomes and interphase nuclei from root tip meristematic cells were obtained by the squashing technique and solid stained for microscope analysis. The DNA amount was estimated by flow cytometry. There were previous reports on the chromosome number of E. edulis and E. oleracea, but chromosome morphology of these two species and the whole karyotype of E. precatoria are reported for the first time. The species have 2n=36, a number considered as a pleisomorphic feature in Arecoideae since the modern species, according to floral morphology, have the lowest chromosome number (2n=28 and 2n=30). The three Euterpe species also have the same type of interphase nuclei, classified as semi-reticulate. The species differed on karyotypic formulas, on localization of secondary constriction and genome size. The data suggest that the main forces driving Euterpe karyotype evolution were structural rearrangements, such as inversions and translocations that alter chromosome morphology, and either deletion or amplification that led to changes in chromosome size.

Ultramorphology and histochemistry of fat body cells from last Instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae)

As células do corpo gorduroso de Pachycondyla (=Neoponera) villosa distribuem-se como uma única camada entre a cutícula e o trato digestivo, formando um conjunto de células agrupadas e recobertas por uma fina membrana. Não foram identificados tipos celulares distintos por meio da ultramorfologia, porém a histologia revelou três tipos celulares distintos: os trofócitos, mais abundantes, as células de urato, distribuídas por entre os trofócitos, e os enócitos, menos abundantes que os demais. Os enócitos são comumente observados próximos da cutícula. Histoquimicamente, os trofócitos apresentaram reação positiva para proteínas básicas no núcleo e no citoplasma e reação fortemente positiva nos grânulos citoplasmáticos. O teste para carboidratos foi fortemente positivo em todo o citoplasma, enquanto para os lipídeos observou-se reação positiva nas vesículas citoplasmáticas. em relação às células de urato, estas apresentaram reação positiva para proteínas básicas no núcleo e citoplasma, por entre as vesículas. Essas células não apresentaram reação para o teste de PAS e Sudan Black B. Quanto aos enócitos, estes apresentaram citoplasma fracamente positivo ao PAS e fortemente positivo ao Sudan Black B e para o azul de bromofenol.

Physiological response of American bullfrog tadpoles to stressor conditions of capture and hypoxia

A rã-touro americana (Rana catesbeiana) recentemente denominada Lithobates catesbeianus é cria da com propósito comercial em várias regiões do Brasil. Situações estressantes tais como problemas de manejo, criação inadequada e alterações ambientais com consequente redução da imunidade são comuns em produções intensivas. A avaliação destas situações de estresse permite-nos detectar estes probemas e diminuir as injurias causadas pelo confinamento. O principal objetivo deste estudo foi utilizar os marcadores biológicos de cortisol, glicemia e dados hematológicos para avaliar a resposta de girinos de rã-touro submetidos aos mecanismos estressores de captura e hipóxia. Os animais foram distribuídosem três tratamentos: estresse por captura individual com puçá; estresse por captura em massa com puçá e estresse por captura por escoamento. Os resultados obtidos demostraram não haver diferenças estatisticamente significativas entre os parametros avaliados quando comparou-se os grupos com e sem exposição ao ar (normoxia e hipoxia). Com base nestes resultados pode-se concluir que os estímulos estressores avaliados não foram adequados para alterar os valores plasmáticos dos marcadores biológicos testados. Para o cortisol, isto ocorreu provavelmente em virtude da ação sinérgica deste hormônio e a tiroxina, que induz a metamorfose nestes animais.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

O objetivo deste estudo foi avaliar a resposta fisiológica (cortisol, glicemia e parâmetros sanguineos) de girinos de rã-touro (Lithobates catesbeianus) em diferentes densidades e após exposição aérea. Os animais utilizados no experimento estavam entre os estágios 31 a 39, na fase de pró-metamorfose sendo testados 1 girino/L (Tratamento 1), 5 girinos/L (Tratamento 2) e 10 girinos/L (Tratamento 3), conduzidos em 3 réplicas simultâneas durante 12 dias. O sangue foi retirado por rompimento do vaso caudal na condição de Normóxia - N (tempo zero) e Hipóxia - H (tempo de 15 minutos de exposição ao ar). Foi observado um aumento nos valores de cortisol, aos 4 e 8 dias de exposição aérea retornando aos valores basais ao final do experimento, apesar de não haver diferenças significativas. A glicemia não apresentou diferenças significativas quanto aos estressores aplicados. Os parâmetros hematológicos da série branca, principalmente, o número de linfócitos, neutrófilos e eosinófilos mostraram diferença significativa aos 12 dias de experimentação quando comparados com o momento zero; concluindo-se que, em condições controladas, o adensamento de até 10 girinos/litro e a exposição aérea por 15 minutos não apresentou danos aos girinos de rã-touro durante o período experimental. O padrão de resposta a estes estímulos talvez seja expresso em outro nível hormonal (corticosterona).

Direct Write Fabrication of Waveguides and Interconnects for Optical Printed Wiring Boards

Current copper based circuit technology is becoming a limiting factor in high speed data transfer applications as processors are improving at a faster rate than are developments to increase on board data transfer. One solution is to utilize optical waveguide technology to overcome these bandwidth and loss restrictions. The use of this technology virtually eliminates the heat and cross-talk loss seen in copper circuitry, while also operating at a higher bandwidth. Transitioning current fabrication techniques from small scale laboratory environments to large scale manufacturing presents significant challenges. Optical-to-electrical connections and out-of-plane coupling are significant hurdles in the advancement of optical interconnects. The main goals of this research are the development of direct write material deposition and patterning tools for the fabrication of waveguide systems on large substrates, and the development of out-of-plane coupler components compatible with standard fiber optic cabling. Combining these elements with standard printed circuit boards allows for the fabrication of fully functional optical-electrical-printed-wiring-boards (OEPWBs). A direct dispense tool was designed, assembled, and characterized for the repeatable dispensing of blanket waveguide layers over a range of thicknesses (25-225 µm), eliminating waste material and affording the ability to utilize large substrates. This tool was used to directly dispense multimode waveguide cores which required no UV definition or development. These cores had circular cross sections and were comparable in optical performance to lithographically fabricated square waveguides. Laser direct writing is a non-contact process that allows for the dynamic UV patterning of waveguide material on large substrates, eliminating the need for high resolution masks. A laser direct write tool was designed, assembled, and characterized for direct write patterning waveguides that were comparable in quality to those produced using standard lithographic practices (0.047 dB/cm loss for laser written waveguides compared to 0.043 dB/cm for lithographic waveguides). Straight waveguides, and waveguide turns were patterned at multimode and single mode sizes, and the process was characterized and documented. Support structures such as angled reflectors and vertical posts were produced, showing the versatility of the laser direct write tool. Commercially available components were implanted into the optical layer for out-of-plane routing of the optical signals. These devices featured spherical lenses on the input and output sides of a total internal reflection (TIR) mirror, as well as alignment pins compatible with standard MT design. Fully functional OEPWBs were fabricated featuring input and output out-of-plane optical signal routing with total optical losses not exceeding 10 dB. These prototypes survived thermal cycling (-40°C to 85°C) and humidity exposure (95±4% humidity), showing minimal degradation in optical performance. Operational failure occurred after environmental aging life testing at 110°C for 216 hours.