Cytotoxicity and Membrane Interaction of Tamoxifen as Affected by Ca2+ and Mg2+: Use of a Bacterial Model System

A strain of Bacillus stearothermophilus was used as a model to study the interaction of tamoxifen (TAM) with the membrane and the cytostatic antiproliferative effects not related to estrogen binding. TAM inhibits the growth of B. stearothermophilus as a function of concentration. The supplementation of the growth medium with Ca2+ or Mg2+ partially relieves the growth inhibition by TAM, allowing growth at TAM concentrations that fully impair growth in the basal medium. Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and of its propionic acid derivative (DPH-PA) reveals opposite effects induced by TAM and Ca2+. The addition of Ca2+ to liposomes of bacterial lipids promoted physical ordering as opposed to disordering induced by TAM. Thus, it is predictable that growth impairment induced by TAM is mediated through perturbations at the membrane level.http://www.sciencedirect.com/science/article/B6TCP-3X3K8KK-Y/1/c91ad40c71c69da7245d8f09c634d28

Seasonal proteome variation in intertidal shrimps under a natural setting: connecting molecular networks with environmental fluctuations

The ability of intertidal organisms to maintain their performance via molecular and physiological adjustments under low tide, seasonal fluctuations and extreme events ultimately determines population viability. Analyzing this capacity in the wild is extremely relevant since intertidal communities are under increased climate variability owing to global changes. We addressed the seasonal proteome signatures of a key intertidal species, the shrimp Palaemon elegans, in a natural setting. Shrimps were collected during spring and summer seasons at low tides and were euthanized in situ. Environmental variability was also assessed using hand-held devices and data loggers. Muscle samples were taken for 2D gel electrophoresis and protein identification through mass spectrometry. Proteome data revealed that 55 proteins (10.6% of the proteome) significantly changed between spring and summer collected shrimps, 24 of which were identified. These proteins were mostly involved in cytoskeleton remodelling, energy metabolism and transcription regulation. Overall, shrimps modulate gene expression leading to metabolic and structural adjustments related to seasonal differences in the wild (i.e. abiotic variation and possibly intrinsic cycles of reproduction and growth). This potentially promotes performance and fitness as suggested by the higher condition index in summer-collected shrimps. However, inter-individual variation (% coefficient of variation) in protein levels was quite low (min-max ranges were 0.6-8.3% in spring and 1.2-4.8% in summer), possibly suggesting reduced genetic diversity or physiological canalization. Protein plasticity is relevant to cope with present and upcoming environmental variation related to anthropogenic forcing (e.g. global change, pollution) but low inter-individual variation may limit evolutionary potential of shrimp populations.publishe

Comparative study of tributyltin toxicity on two bacteria of the genus Bacillus

Tributyltin is a potent biocide mainly used in marine antifouling paints. Owing to its widespread distribution in coast areas and its high toxicity to aquatic organisms, the use of this compound is generally restricted and under government regulation. Despite of that, it persists in the aquatic environment. Organotins used in industry have also been detected in terrestrial environments. The persistence and high lipophilicity explain bioaccumulation. The role of bacteria in recycling organic matter prompted us to study the interaction of tributyltin with two ubiquitous bacilli, B. stearothermophilus and B. subtilis, proposed as biological indicators of pollutants with ecological impact. These bacteria have been used as suitable models for the study of toxicity mechanisms of unselective lipophilic compounds (e.g., DDT and endosulfan). Drug effects on growth parameters, oxygen consumption and membrane organization were assessed. Bacteria growth in a liquid complex medium was disturbed by concentrations of TBT as low as 25 nM (8 [mu]g L-1), close to the concentration in polluted environments. The respiratory activity is affected by TBT in both microorganisms. Membrane organization, assessed by fluorescence polarization of two fluidity probes, 1,6-diphenyl-1,3,5-hexatriene (DPH) and a propionic acid derivative (DPH-PA), was also perturbed by the xenobiotic. Alterations on growth, oxygen consumption and physical properties of membrane lipids are stronger in B. stearothermophilus as compared to B. subtilis. A putative relationship between growth inhibition and respiratory activity impairment induced by TBT and its effects on the physical behaviour of bacterial membrane lipids is suggested.http://www.sciencedirect.com/science/article/B6TCP-4GSJR3R-9/1/18e0ee5c8ce4e887466d84b850ad8ec

Conjugated linoleic acid reduces permeability and fluidity of adipose plasma membranes from obese Zucker rats

NOTICE: this is the author’s version of a work that was accepted for publication in Biochemical and Biophysical Research Communications. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Biochemical and Biophysical Research Communications. July 2010; 398 (2): 199-204.Conjugated linoleic acid (CLA) is a dietary fatty acid frequently used as a body fat reducing agent whose effects upon cell membranes and cellular function remain unknown. Obese Zucker rats were fed atherogenic diets containing saturated fats of vegetable or animal origin with or without 1% CLA, as a mixture of cis(c)9,trans(t)11 and t10,c12 isomers. Plasma membrane vesicles obtained from visceral adi- pose tissue were used to assess the effectiveness of dietary fat and CLA membrane incorporation and its outcome on fluidity and permeability to water and glycerol. A significant decrease in adipose membrane fluidity was correlated with the changes observed in permeability, which seem to be caused by the incor- poration of the t10,c12 CLA isomer into membrane phospholipids. These results indicate that CLA supple- mentation in obese Zucker rats fed saturated and cholesterol rich diets reduces the fluidity and permeability of adipose membranes, therefore not supporting CLA as a body fat reducing agent through membrane fluidification in obese fat consumers

Toxicity of methoprene as assessed by the use of a model microorganism

Methoprene is an insect juvenile growth hormone mimic, commonly used as a pesticide. Although widely used for the control of several pests, toxic effects on organisms of different phyla have been reported. These events triggered studies to clarify the mechanisms of toxicity of this insecticide putatively involved in ecological issues. Here we show the effect of methoprene on the normal cell growth and viability of a strain of the thermophilic eubacterium Bacillus stearothermophilus, previously used as a model for toxicological evaluation of other environment pollutants. Respiration studies were also carried out attempting to identify a putative target for the cytotoxic action of methoprene. Cell growth was affected and a decrease of the number of viable cells was observed as a result of the addition of methoprene to the growth medium, an effect reverted by the presence of Ca2+. Methoprene also inhibited the redox flow of B. stearothermophilus protoplasts before the cytochrome oxidase segment, an effect further studied by individually assessing the enzymatic activities of the respiratory complexes. This study suggests that methoprene membrane interaction and perturbation of cell bioenergetics may underlie the mechanism of toxicity of this compound in non-target organisms.http://www.sciencedirect.com/science/article/B6TCP-4GSJXGW-4/1/788f9fd7d4f3a4f1ae5d498a5674dcc

Hydroxytamoxifen interaction with human erythrocyte membrane and induction of permeabilization and subsequent hemolysis

4-Hydroxytamoxifen (OHTAM) is the most active metabolite of the widely prescribed anticancer drug tamoxifen (TAM) used in breast cancer therapy. This work describes the effects of OHTAM on isolated human erythrocytes, using standardized test conditions, to check for a putative contribution to the TAM-induced hemolysis and to study basic mechanisms involved in the interaction of OHTAM with cell membranes. Incubation of isolated human erythrocytes with relatively high concentrations of OHTAM results in a concentration-dependent hemolysis, its hemolytic effect being about one-third of that induced by TAM. OHTAM-induced hemolysis is prevented by either [alpha]-tocopherol ([alpha]-T) or [alpha]-tocopherol acetate ([alpha]-TAc) and it occurs in the absence of oxygen consumption and hemoglobin oxidation, ruling out the oxidative damage of erythrocytes. However, OHTAM remarkably increases the osmotic fragility of erythrocytes, increasing the susceptibility of erythrocytes to hypotonic lysis. Additionally, the hemoglobin release induced by OHTAM is preceded by a rapid efflux of intracellular K+. Therefore, our data suggest that OHTAM-induced hemolysis does not contribute to TAM-induced hemolytic anemia and it is a much weaker toxic drug as compared with TAM. Moreover, at variance with the membrane disrupting effects of TAM, OHTAM promotes perturbation of the membrane's backbone region due to its strong binding to proteins with consequent formation of membrane paths of permeability to small solutes and retention of large solutes like hemoglobin, followed by osmotic swelling and cell lysis. The prevention of OHTAM-induced hemolysis by [alpha]-T and [alpha]-TAc is probably committed to the permeability sealing resulting from structural stabilization of membrane.http://www.sciencedirect.com/science/article/B6TCP-44B2835-2/1/29f8930b906a9b147f7df74b698fdbb

Does harvest residue management influence biomass and nutrient accumulation in understory vegetation of Eucalyptus globulus Labill. plantations in a Mediterranean environment ?

The effect of harvest residue management options on biomass and nutrient accumulation in understory vegetation, as well as the contribution of understory to nutrient cycling, were assessed during the early rotation stage of a Eucalyptus globulus Labill. plantation in Central Portugal. The effects of residue management options on early tree growth were also evaluated. Treatments established at the time of plantation and replicated four times in a simple completely randomised design included removal of harvest residues (R), incorporation of residues into the soil by harrowing (I) andmaintenance of residues on the soil surface (S). Understory biomass was sampled in the spring between 2002 and 2006, and every 2 months between March 2006 and March 2007. The latter samples were stratified into biomass, standing dead mass and litter for net above ground primary production (NAPP) assessment. Samples were oven dried, weighed and analysed for nutrient contents. Results showed that understory standing biomass strongly increased from the first to the third year and that quantities of nutrients accumulated in ground vegetation followed similar patterns between the three treatments. Nutrient accumulation in ground vegetation was greater than in tree biomass until at least the second spring after plantation. Bimonthly sampling revealed treatment R to have the largest amounts of standing biomass, standing dead mass, litter and nutrient immobilisation, while treatment S exhibited the lowest values. NAPP (4th–5th year) was 639, 511 and 362 g m 2 year 1, respectively in R, I and S, corresponding the standing biomass increase to 277, 183 and 143 g m 2 year 1. These values are comparable to those observed for litter fall in similar stands (age and tree density) in the same area. The contribution of ground vegetation to nutrient accumulation in the system was unaffected by harvest residue management methods, but further research is necessary in order to establish whether slash management options influence long term tree growth and vegetation dynamics

Momentum distribution of Vinen turbulence in trapped atomic Bose-Einstein condensates

The decay of multicharged vortices in trapped Bose-Einstein condensates may lead to a disordered vortex state consistent with the Vinen regime of turbulence, characterized by an absence of large-scale flow and an incompressible kinetic energy spectrum $E\propto k^{-1}$. In this work, we study numerically the dynamics of a three-dimensional harmonically trapped Bose-Einstein condensate excited to a Vinen regime of turbulence through the decay of two doubly-charged vortices. First, we study the momentum distribution and observe the emergence of a power-law behavior $n(k)\propto k^{-3}$ consistent with the coexistence of wave turbulence. We also study the kinetic energy and particle fluxes, which allows us to identify a direct particle cascade associated with the turbulent stage.Comment: 5 pages, 2 figure