c‐FLIP is involved in erythropoietin‐mediated protection of erythroid‐differentiated cells from TNF‐α‐induced apoptosis

The TNF‐α (tumour necrosis factor) affects a wide range of biological activities, such as cell proliferation and apoptosis. Cell life or death responses to this cytokine might depend on cell conditions. This study focused on the modulation of factors that would affect the sensitivity of erythroid‐differentiated cells to TNF‐α. Hemin‐differentiated K562 cells showed higher sensitivity to TNF‐induced apoptosis than undifferentiated cells. At the same time, hemin‐induced erythroid differentiation reduced c‐FLIP (cellular FLICE‐inhibitory protein) expression. However, this negative effect was prevented by prior treatment with Epo (erythropoietin), which allowed the cell line to maintain c‐FLIP levels. On the other hand, erythroid‐differentiated UT‐7 cells – dependent on Epo for survival – showed resistance to TNF‐α pro‐apoptotic action. Only after the inhibition of PI3K (phosphatidylinositol‐3 kinase)‐mediated pathways, which was accompanied by negative c‐FLIP modulation and increased erythroid differentiation, were UT‐7 cells sensitive to TNF‐α‐triggered apoptosis. In summary, erythroid differentiation might deregulate the balance between growth promotion and death signals induced by TNF‐α, depending on cell type and environmental conditions. The role of c‐FLIP seemed to be critical in the protection of erythroid‐differentiated cells from apoptosis or in the determination of their sensitivity to TNF‐mediated programmed cell death. Epo, which for the first time was found to be involved in the prevention of c‐FLIP down‐regulation, proved to have an anti‐apoptotic effect against the pro‐inflammatory factor. The identification of signals related to cell life/death switching would have significant implications in the control of proliferative diseases and would contribute to the understanding of mechanisms underlying the anaemia associated with inflammatory processes.     

Quantitative 3D fluorescence technique for the analysis of en face preparations of arterial walls using quantum dot nanocrystals and two-photon excitation laser scanning microscopy

Traditional imaging with one-photon confocal microscopy and organic fluorophores poses several challenges for the visualization of vascular tissue, including autofluorescence, fluorophore crosstalk, and photobleaching. We studied human coronary arteries (HCAs) and mouse aortas with a modified immunohistochemical (IHC) "en face" method using quantum dot (Qdot) bioconjugates and two-photon excitation laser scanning microscopy (TPELSM). We demonstrated the feasibility of multilabeling intimal structures by exciting multicolored Qdots with only one laser wavelength (750 nm). Detailed cell structures, such as the granular appearance of von Willebrand factor (VWF) and the subcellular distribution of endothelial nitric oxide synthase, were visualized using green dots (525 nm), even when the emission maximum of these Qdots overlapped that of tissue autofluorescence (510-520 nm). In addition, sensitive fluorescence quantification of vascular cell adhesion molecule 1 expression at areas of varying hemodynamics (intercostal branches vs. nonbranching areas) was performed in normal C57Bl/6 mice. Finally, we took advantage of the photostability of Qdots and the inherent three-dimensional (3D) resolution of TPELSM to obtain large z-stack series without photobleaching. This innovative en face method allowed simple multicolor profiling, highly sensitive fluorescence quantitation, and 3D visualization of the vascular endothelium with excellent spatial resolution. This is a promising technique to define the spatial and temporal interactions of endothelial inflammatory markers and quantify the effects of different interventions on the endothelium.     

Differential antiapoptotic effect of erythropoietin on undifferentiated and retinoic acid‐differentiated SH‐SY5Y cells

Erythropoietin (Epo) is known to have a significant role in tissues outside the hematopoietic system. In this work, we investigated the function of Epo in cells of neuronal origin subjected to differentiation. Treatment of SH‐SY5Y cells with all‐trans‐retinoic acid (atRA) generated differentiated neuron‐like cells, observed by increased expression of neuronal markers and morphological changes. Exposure of undifferentiated cells to proapoptotic stimuli such as staurosporine, TNF‐α, or hypoxia, significantly increased programmed cell death, which was prevented by previous treatment with Epo. In contrast, atRA‐differentiated cultures showed cell resistance to apoptosis. No additional effect of Epo was detected in previously differentiated cells. The inhibition of the PI3K/Akt pathway by Ly294002 abrogated the protective effects induced by either Epo or atRA. The effect of atRA was mediated by an increased expression of Bcl‐2 whereas the Epo treatment upregulated not only Bcl‐2 but also Bcl‐xL. This upregulation by Epo was not detected in atRA‐differentiated cells, thus confirming the lack of the protective effect of Epo. As expected, assays with AG490, an inhibitor of Jak2, blocked the Epo action only in undifferentiated cells. This reduced neuroprotective function of Epo on SH‐SY5Y differentiated cells could be explained at least in part by downregulation of the Epo receptor expression, which was observed in atRA‐differentiated cells. This study shows differential cellular protection induced by Epo at two stages of SH‐SY5Y differentiation. The results allow us to suggest that this differential cell behavior can be ascribed to the interaction between atRA and the signaling pathways mediated by Epo. J. Cell. Biochem. 110: 151–161, 2010. © 2010 Wiley‐Liss, Inc.     

High‐fat diet and hydrochlorothiazide increase oxidative stress in brain of rats

This study evaluated the effect of possible synergic interaction between high fat diet (HF) and hydrochlorothiazide (HCTZ) on biochemical parameters of oxidative stress in brain. Rats were fed for 16 weeks with a control diet or with an HF, both supplemented with different doses of HCTZ (0.4, 1.0, and 4.0 g kg⁻¹ of diet). HF associated with HCTZ caused a significant increase in lipid peroxidation and blood glucose levels. In addition, HF ingestion was associated with an increase in cerebral lipid peroxidation, vitamin C and non‐protein thiol groups (NPSH) levels. There was an increase in vitamin C as well as NPSH levels in HCTZ (1.0 and 4.0 g kg⁻¹ of diet) and HF plus HCTZ groups. Na⁺–K⁺‐ATPase activity of HCTZ (4.0 g kg⁻¹ of diet) and HCTZ plus HF‐fed animals was significantly inhibited. Our data indicate that chronic intake of a high dose of HCTZ (4 g kg⁻¹ of diet) or HF change biochemical indexes of oxidative stress in rat brain. Furthermore, high‐fat diets consumption and HCTZ treatment have interactive effects on brain, showing that a long‐term intake of high‐fat diets can aggravate the toxicity of HCTZ. Copyright © 2009 John Wiley & Sons, Ltd.     

Poverty,sanitation, and Leptospira transmission pathways in residents from four Brazilian slums

Residents of urban slums suffer from a high burden of zoonotic diseases due to individual, socioeconomic, and environmental factors. We conducted a cross-sectional sero-survey in four urban slums in Salvador, Brazil, to characterize how poverty and sanitation contribute to the transmission of rat-borne leptospirosis. Sero-prevalence in the 1,318 participants ranged between 10.0 and 13.3%. We found that contact with environmental sources of contamination, rather than presence of rat reservoirs, is what leads to higher risk for residents living in areas with inadequate sanitation. Further, poorer residents may be exposed away from the household, and ongoing governmental interventions were not associated with lower transmission risk. Residents at higher risk were aware of their vulnerability, and their efforts improved the physical environment near their household, but did not reduce their infection chances. This study highlights the importance of understanding the socioeconomic and environmental determinants of risk, which ought to guide intervention efforts.     

Hotspots of annual fish along the Patos-Mirim Lagoon System and adjacent areas in Southern Brazil,with an evaluation of conservation priority for species and localities

Biodiversity and Conservation - Rivulidae represents one of the most endangered taxa in Brazil. This family encompasses species known as annual or seasonal fish because they possess unique...     

Linking Microbial Community and Catabolic Gene Structures during the Adaptation of Three Contaminated Soils under Continuous Long-Term Pollutant Stress

Three types of contaminated soil from three geographically different areas were subjected to a constant supply of benzene or benzene/toluene/ethylbenzene/xylenes (BTEX) for a period of 3 months. Different from the soil from Brazil (BRA) and Switzerland (SUI), the Czech Republic (CZE) soil which was previously subjected to intensive in situ bioremediation displayed only negligible changes in community structure. BRA and SUI soil samples showed a clear succession of phylotypes. A rapid response to benzene stress was observed, whereas the response to BTEX pollution was significantly slower. After extended incubation, actinobacterial phylotypes increased in relative abundance, indicating their superior fitness to pollution stress. Commonalities but also differences in the phylotypes were observed. Catabolic gene surveys confirmed the enrichment of actinobacteria by identifying the increase of actinobacterial genes involved in the degradation of pollutants. Proteobacterial phylotypes increased in relative abundance in SUI microcosms after short-term stress with benzene, and catabolic gene surveys indicated enriched metabolic routes. Interestingly, CZE soil, despite staying constant in community structure, showed a change in the catabolic gene structure. This indicates that a highly adapted community, which had to adjust its gene pool to meet novel challenges, has been enriched.     

Influence of macroalgal morphology on the functional structure of molluscan community from hypersaline estuary

Hydrobiologia - Studies based on functional approach seek to understand the ecological roles developed by species as well as their interactions with the environment in which they are inserted. The...     

Indirect effect of elevated CO2 concentration on Bemisia tabaci MEAM1 feeding on Bt soybean plants

The development of herbivore insects is influenced by the quality of their host plants. Elevated CO₂ alters plant metabolism, which may change the nutritional quality of the plant, modifying the life history and feeding behaviour of herbivore insects. Understanding how insect pests respond to increasing CO₂ concentration is essential for predicting the impact of the pest on food security. In this study, we investigated the effects of elevated CO₂ (eCO₂) on the life history and feeding behaviour of the MEAM1 species of Bemisia tabaci on a Bt soybean cultivar. We found that eCO₂ increased the egg to adult development time and reduced the reproductive responses (fecundity and fertility) of B. tabaci. The whitefly B. tabaci that fed on the soybean plants grown under eCO₂ conditions was negatively influenced by several traits related to the host plant resistance, such as the time spent on phloem sap ingestion. Furthermore, we evaluated the changes in the C:N concentration and plant morphology of the Bt plants. The biomass (weight of leaves and stems) of the Bt soybean plants grown under eCO₂ conditions was significantly increased, and the elevated C:N ratio in the phenological stage V6 (i.e. when the plants had six trifoliate leaves developed) was the most pronounced difference in the Bt soybean plants subjected to eCO₂ treatment. Taken together, our results indicate that Bt plants cultivated under eCO₂ inhibit B. tabaci feeding, which can reduce whitefly infestations of the soybean fields.     

A multilocus perspective on the phylogenetic relationships and diversification of rodents of the tribe Abrotrichini (Cricetidae: Sigmodontinae)

Abrotrichini is a recently defined and diagnosed tribe of Sigmodontinae with a complex taxonomy. Abrotrichine genera, Abrothrix (including Chroeomys), Chelemys, Geoxus, Notiomys and Pearsonomys, are mostly distributed in the central and southern Andes and adjacent lowlands and show terrestrial and fossorial habits. Recent studies have evidenced some incongruence between current taxonomy and abrotrichine phylogeny, such as the polyphyly of Chelemys and paraphyly of Geoxus respect to Pearsonomys. We used DNA sequence data of six loci (one mitochondrial and five nuclear) to resolve the relationships within the tribe. Independent and combined analyses of these loci were carried out using parsimony, maximum likelihood and Bayesian inference. Estimates of divergence time of the main lineages of abrotrichines were calculated with a molecular clock using as calibration, a fossil recently found. The concatenated data set increased the resolution and defined the relationships within the tribe. Our phylogenetic analyses corroborate that Abrothrix is sister of a clade containing the long‐clawed abrotrichines. We recovered two main clades within Abrothrix that match morphologic variation and geographic distribution of its species. In addition, we corroborated the lack of monophyly of Chelemys and the lack of monophyly of Geoxus. We discuss different taxonomic scenarios to abrotrichine classification reflects the phylogenetic relationships obtained in this study. Our molecular clock estimated the Abrotrichini crown age to be around the early Pliocene (4.4 Ma) and suggest that the tribe diversified over a short period of time.