Arsenic bioaccumulation and biotransformation in different tissues of Nile tilapia (Oreochromis niloticus): A comparative study between As(III) and As(V) exposure and evaluation of antagonistic effects of selenium.

The speciation of arsenic in fish has been widely investigated, but bioaccumulation and biotransformation of inorganic As in different tissues of Nile tilapia (Oreochromis niloticus) are not fully understood. The present study aimed to investigate the bioaccumulation of As in Nile tilapia, as well as to evaluate the distribution of the main arsenic species (As(III), As(V), MMA, DMA, and AsB) in liver, stomach, gill, and muscle, after controlled exposures to As(III) and As(V) at concentrations of 5.0 and 10.0 mg L-1 during periods of 1 and 7 days. Total As was determined by inductively coupled plasma mass spectroscopy (ICP-MS). For both exposures (As(III) and As(V)), the total As levels after 7-day exposure were highest in the liver and lowest in the muscle. Overall, the Nile tilapia exposed to As(III) showed higher tissue levels of As after the treatments, compared to As(V) exposure. Speciation of arsenic present in the tissues employed liquid chromatography coupled to ICP-MS (LC-ICP-MS), revealing that the biotransformation of As included As(V) reduction to As(III), methylation to monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA), and subsequent conversion to nontoxic arsenobetaine (AsB), which was the predominant arsenic form. Finally, the interactions and antagonistic effects of selenium in the bioaccumulation processes were tested by the combined exposure to As(III), the most toxic species of As, together with tetravalent selenium (Se(IV)). The results indicated a 4-6 times reduction of arsenic toxicity in the tilapia.

Peroxiredoxins in erythrocytes: far beyond the antioxidant role.

The red blood cells (RBCs) are essential to transport oxygen (O2) and nutrients throughout the human body. Changes in the structure or functioning of the erythrocytes can lead to several deficiencies, such as hemolytic anemias, in which an increase in reactive oxidative species generation is involved in the pathophysiological process, playing a significant role in the severity of several clinical manifestations. There are important lines of defense against the damage caused by oxidizing molecules. Among the antioxidant molecules, the enzyme peroxiredoxin (Prx) has the higher decomposition power of hydrogen peroxide, especially in RBCs, standing out because of its abundance. This review aimed to present the recent findings that broke some paradigms regarding the three isoforms of Prxs found in RBC (Prx1, Prx2, and Prx6), showing that in addition to their antioxidant activity, these enzymes may have supplementary roles in transducing peroxide signals, as molecular chaperones, protecting from membrane damage, and maintenance of iron homeostasis, thus contributing to the overall survival of human RBCs, roles that seen to be disrupted in hemolytic anemia conditions.

FoxO3 and oxidative stress: a multifaceted role in cellular adaptation.

Oxidative stress is a major cause of morbidity and mortality in human health and disease. In this review, we focus on the Forkhead Box (Fox) subclass O3 (FoxO3), an extensively studied transcription factor that plays a pleiotropic role in a wide range of physiological and pathological processes by regulating multiple gene regulatory networks involved in the modulation of numerous aspects of cellular metabolism, including fuel metabolism, cell death, and stress resistance. This review will also focus on regulatory mechanisms of FoxO3 expression and activity, such as crucial post-translational modifications and non-coding RNAs. Moreover, this work discusses and evidences some pathways to how this transcription factor and reactive oxygen species regulate each other, which may lead to the pathogenesis of various types of diseases. Therefore, in addition to being a promising therapeutic target, the FoxO3-regulated signaling pathways can also be used as reliable diagnostic and prognostic biomarkers and indicators for drug responsiveness.

Influence of Melatonin Treatment on Cellular Mechanisms of Redox Adaptation in K562 Erythroleukemic Cells.

Melatonin (MEL) presents well-documented pleiotropic actions against oxidative stress (OS), acting indirectly through activation of transcription factors, e.g., FoxO3 and Nrf2. Thus, this study aimed to investigate the possible modulating effects of MEL on the redox signaling pathways PI3K/AKT/FoxO3 and Keap1/Nrf2/ARE in K562 erythroleukemic cells subjected to OS induction. For this, the viability, and transcript levels of genes involved in redox adaptation were evaluated in K562 cells in different periods of erythroid differentiation: under OS induction by hydrogen peroxide (100 µM H2O2); treated with 1 nM (C1) and 1 mM (C2) MEL; and associated or not with stress induction. We observed a restoration of physiological levels of Nrf2 in both MEL concentrations under OS. The C1 was related to enhanced expression of antioxidant and proteasome genes through the Nrf2-ARE pathway, while C2 to the induction of FOXO3 expression, suggesting an involvement with apoptotic pathway, according to BIM transcript levels. The effects of MEL administration in these cells showed a period and dose-dependent pattern against induced-OS, with direct and indirect actions through different pathways of cellular adaptation, reinforcing the importance of this indolamine in the regulation of cellular homeostasis, being a promising therapeutic alternative for diseases that present an exacerbated OS.

Potential Cytoprotective and Regulatory Effects of Ergothioneine on Gene Expression of Proteins Involved in Erythroid Adaptation Mechanisms and Redox Pathways in K562 Cells.

This study aimed to establish the importance of ergothioneine (ERT) in the erythroid adaptation mechanisms by appraising the expression levels of redox-related genes associated with the PI3K/AKT/FoxO3 and Nrf2-ARE pathways using K562 cells induced to erythroid differentiation and H2O2-oxidative stress. Cell viability and gene expression were evaluated. Two concentrations of ERT were assessed, 1 nM (C1) and 100 µM (C2), with and without stress induction (100 µM H2O2). Assessments were made in three periods of the cellular differentiation process (D0, D2, and D4). The C1 treatment promoted the induction of FOXO3 (D0 and 2), PSMB5, and 6 expressions (D4); C1 + H2O2 treatment showed the highest levels of NRF2 transcripts, KEAP1 (D0), YWHAQ (D2 and 4), PSMB5 (D2) and PSMB6 (D4); and C2 + H2O2 (D2) an increase in FOXO3 and MST1 expression, with a decrease of YWHAQ and NRF2 was observed. in C2 + H2O2 (D2) an increase in FOXO3 and MST1, with a decrease in YWHAQ and NRF2 was observed All ERT treatments increased gamma-globin expression. Statistical multivariate analyzes highlighted that the Nrf2-ARE pathway presented a greater contribution in the production of PRDX1, SOD1, CAT, and PSBM5 mRNAs, whereas the PI3K/AKT/FoxO3 pathway was associated with the PRDX2 and TRX transcripts. In conclusion, ERT presented a cytoprotective action through Nrf2 and FoxO3, with the latter seeming to contribute to erythroid proliferation/differentiation.

Association of FOXO3 polymorphism (rs3800231) and clinical subphenotypes of beta thalassemic individuals

Abstract Introduction Studies have shown that the loss of the FOXO3 transcriptional function is involved in the pathophysiology of some chronic erythroid disorders, including beta-thalassemia (β-thal). Therefore, the single nucleotide polymorphism (SNP) rs3800231 (35-2764A > G) could contribute to alterations in its transcriptional activity, acting as a modifier of β-thal phenotypic manifestations. Objective and method In order to better understand the genotypic and/or allelic distributions among β-thal patients, we evaluated 83 β-thal heterozygous and 20 homozygous, compared to 117 individuals without hemoglobinopathies (control group). Additionally, we verified any influence of the FOXO3 polymorphism on clinical manifestations among β-thal homozygotes. Results We obtained higher frequencies of the wild-type homozygous (AA) and the wild-type allele (A) in the β-thal group (p< 0.0001 and p= 0.00014, respectively). The most common clinical manifestations found among β-thal homozygotes were iron overload (90%), splenomegaly (65%) and bone complications (35%), e.g., osteopenia/osteoporosis. We observed that close to 80% of the patients presenting such manifestations had the genotype AA. However, we did not find any significant involvement of the FOXO3 polymorphism in clinical manifestation occurrences. Conclusion Thus, we concluded that the SNP rs3800231 did not play a significant role as a modifier of the clinical manifestations observed in the β-thal homozygotes studied.

Association of FOXO3 polymorphism (rs3800231) and clinical subphenotypes of beta thalassemic individuals.

INTRODUCTION: Studies have shown that the loss of the FOXO3 transcriptional function is involved in the pathophysiology of some chronic erythroid disorders, including beta-thalassemia (ß-thal). Therefore, the single nucleotide polymorphism (SNP) rs3800231 (35-2764A > G) could contribute to alterations in its transcriptional activity, acting as a modifier of ß-thal phenotypic manifestations. OBJECTIVE AND METHOD: In order to better understand the genotypic and/or allelic distributions among ß-thal patients, we evaluated 83 ß-thal heterozygous and 20 homozygous, compared to 117 individuals without hemoglobinopathies (control group). Additionally, we verified any influence of the FOXO3 polymorphism on clinical manifestations among ß-thal homozygotes. RESULTS: We obtained higher frequencies of the wild-type homozygous (AA) and the wild-type allele (A) in the ß-thal group (p < 0.0001 and p = 0.00014, respectively). The most common clinical manifestations found among ß-thal homozygotes were iron overload (90%), splenomegaly (65%) and bone complications (35%), e.g., osteopenia/osteoporosis. We observed that close to 80% of the patients presenting such manifestations had the genotype AA. However, we did not find any significant involvement of the FOXO3 polymorphism in clinical manifestation occurrences. CONCLUSION: Thus, we concluded that the SNP rs3800231 did not play a significant role as a modifier of the clinical manifestations observed in the ß-thal homozygotes studied.

Nanostructure of PMMA/MAM Blends Prepared by Out-of-Equilibrium (Extrusion) and Near-Equilibrium (Casting) Self-Assembly and Their Nanocellular or Microcellular Structure Obtained from CO2 Foaming.

Blends of poly(methyl methacrylate) (PMMA) and a triblock copolymer poly(methyl methacrylate)-b-poly(butyl acrylate)-b-poly(methyl methacrylate) (MAM) have been obtained following both out-of-equilibrium (extrusion) and near-equilibrium (solvent casting) production routes. The self-assembly capability and the achievable nanostructures of these blends are analyzed by transmission electron microscopy (TEM) regarding their production route and potential for the achievement of nanocellular foams by CO2 gas dissolution foaming. The influence of the initial nanostructure of the solids on the obtained cellular structure of bulk and film samples is determined by high-resolution scanning electron microscopy (HRSEM) for diverse foaming conditions (saturation pressure, saturation temperature, and post-foaming stage), taking into account the required use of a foaming mold to achieve foams from films. Moreover, the influence of the nanostructuration on the presence of solid outer layers, typical of the selected foaming process, is addressed. Finally, consideration of a qualitative model and the obtained results in terms of nanostructuration, cellular structure, and foaming behavior, allow proposing a detailed cell nucleation, growth, and stabilization scheme for these materials, providing the first direct evidence of the cell nucleation happening inside the poly(butyl acrylate) phase in the PMMA/MAM blends.

Effect of the Molecular Structure of TPU on the Cellular Structure of Nanocellular Polymers Based on PMMA/TPU Blends.

In this work, the effects of thermoplastic polyurethane (TPU) chemistry and concentration on the cellular structure of nanocellular polymers based on poly(methyl-methacrylate) (PMMA) are presented. Three grades of TPU with different fractions of hard segments (HS) (60%, 70%, and 80%) have been synthesized by the prepolymer method. Nanocellular polymers based on PMMA have been produced by gas dissolution foaming using TPU as a nucleating agent in different contents (0.5 wt%, 2 wt%, and 5 wt%). TPU characterization shows that as the content of HS increases, the density, hardness, and molecular weight of the TPU are higher. PMMA/TPU cellular materials show a gradient cell size distribution from the edge of the sample towards the nanocellular core. In the core region, the addition of TPU has a strong nucleating effect in PMMA. Core structure depends on the HS content and the TPU content. As the HS or TPU content increases, the cell nucleation density increases, and the cell size is reduced. Then, the use of TPUs with different characteristics allows controlling the cellular structure. Nanocellular polymers have been obtained with a core relative density between 0.15 and 0.20 and cell sizes between 220 and 640 nm.

Analysis of the Foaming Window for Thermoplastic Polyurethane with Different Hard Segment Contents.

A series of thermoplastic polyurethanes (TPUs) with different amounts of hard segments (HS) (40, 50 and 60 wt.%) are synthesized by a pre-polymer method. These synthesized TPUs are characterized by Shore hardness, gel permeation chromatography (GPC), differential scanning calorimetry (DSC), wide angle X-ray diffraction (WAXD), dynamic mechanical thermal analysis (DMTA), and rheology. Then, these materials are foamed by a one-step gas dissolution foaming process and the processing window that allows producing homogeneous foams is analyzed. The effect of foaming temperature from 140 to 180 °C on the cellular structure and on density is evaluated, fixing a saturation pressure of 20 MPa and a saturation time of 1 h. Among the TPUs studied, only that with 50 wt.% HS allows obtaining a stable foam, whose better features are reached after foaming at 170 °C. Finally, the foaming of TPU with 50 wt.% HS is optimized by varying the saturation pressure from 10 to 25 MPa at 170 °C. The optimum saturation and foaming conditions are 25 MPa and 170 °C for 1 h, which gives foams with the lowest relative density of 0.74, the smallest average cell size of 4 µm, and the higher cell nucleation density of 8.0 × 109 nuclei/cm3. As a final conclusion of this investigation, the TPU with 50 wt.% HS is the only one that can be foamed under the saturation and foaming conditions used in this study. TPU foams containing 50 wt.% HS with a cell size below 15 microns and porosity of 1.4-18.6% can be obtained using foaming temperatures from 140 to 180 °C, saturation pressure of 20 MPa, and saturation time of 1 h. Varying the saturation pressure from 10 to 25 MPa and fixing the foaming temperature of 170 °C and saturation pressure of 1 h results in TPU foams with a cell size of below 37 microns and porosity of 1.7-21.2%.

Cyclic Gas Dissolution Foaming as an Approach for Simultaneously Reducing Cell Size and Relative Density in Nanocellular PMMA.

A new approach to produce nanocellular polymers combining small cell sizes with low relative densities is presented herein. This production method, based on gas dissolution foaming, consists of performing a double saturation and foaming cycle. Thus, nanocellular polymethylmethacrylate (PMMA) has been produced through a first saturation at different saturation conditions (6, 10, and 20 MPa and -32 °C), at constant foaming conditions (60 °C for 1 min). Then, the nanocellular PMMAs obtained from the previous step were again saturated at different saturation conditions, 10 MPa 24 °C, 31 MPa 24 °C, 35 MPa 22 °C, and 6 MPa -15 °C and foamed at different temperatures (40, 80 and 100 °C) for 1 min. This new approach allows the cells created in the first saturation and foaming cycle to further grow in the second cycle. This fact permits producing nanocellular polymethylmethacrylate sheets combining, for the first time in the literature, cell sizes of 24 nm with relative densities of 0.3.

The mechanics of solid-state nanofoaming.

Solid-state nanofoaming experiments are conducted on two polymethyl methacrylate (PMMA) grades of markedly different molecular weight using CO2 as the blowing agent. The sensitivity of porosity to foaming time and foaming temperature is measured. Also, the microstructure of the PMMA nanofoams is characterized in terms of cell size and cell nucleation density. A one-dimensional numerical model is developed to predict the growth of spherical, gas-filled voids during the solid-state foaming process. Diffusion of CO2 within the PMMA matrix is sufficiently rapid for the concentration of CO2 to remain almost uniform spatially. The foaming model makes use of experimentally calibrated constitutive laws for the uniaxial stress versus strain response of the PMMA grades as a function of strain rate and temperature, and the effect of dissolved CO2 is accounted for by a shift in the glass transition temperature of the PMMA. The maximum achievable porosity is interpreted in terms of cell wall tearing and comparisons are made between the predictions of the model and nanofoaming measurements; it is deduced that the failure strain of the cell walls is sensitive to cell wall thickness.

Nanocellular Polymers: The Challenge of Creating Cells in the Nanoscale.

The evolution of technology means that increasingly better materials are needed. It is well known that as a result of their interesting properties, nanocellular polymers perform better than microcellular ones. For this reason, the investigation on nanocellular materials is nowadays a very topical issue. In this paper, the different approaches for the production of these materials in our laboratory are explained, and results obtained by using polymethylmethacrylate (PMMA) are shown. Homogeneous nucleation has been studied by using raw PMMA, while two different systems were used for heterogeneous nucleation; adding nanoparticles to the system and using nanostructured polymers as solid precursors for foaming. The effects of the different parameters of the production process (gas dissolution foaming process) have been evaluated for all systems being possible to establish a comparison between the materials produced by different approaches. Moreover, the limitations and future work to optimise the materials produced are also discussed.

Nanoclay Intercalation During Foaming of Polymeric Nanocomposites Studied in-Situ by Synchrotron X-Ray Diffraction.

The intercalation degree of nanoclays in polymeric foamed nanocomposites containing clays is a key parameter determining the final properties of the material, but how intercalation occurs is not fully understood. In this work, energy dispersive X-ray diffraction (ED-XRD) of synchrotron radiation was used as an in-situ technique to deepen into the intercalation process of polymer/nanoclay nanocomposites during foaming. Foamable nanocomposites were prepared by the melt blending route using low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS) with surface treated nanoclays and azodicarbonamide (ADC) as the blowing agent. Foaming was induced by heating at atmospheric pressure. The time and temperature evolution of the interlamellar distance of the clay platelets in the expanding nanocomposites was followed. Upon foaming, interlamellar distances of the nanocomposites based on LDPE and PP increase by 18% and 16% compared to the bulk foamable nanocomposite. Therefore, the foaming process enhances the nanoclay intercalation degree in these systems. This effect is not strongly affected by the type of nanoclay used in LDPE, but by the type of polymer used. Besides, the addition of nanoclays to PP and PS has a catalytic effect on the decomposition of ADC, i.e., the decomposition temperature is reduced, and the amount of gas released increases. This effect was previously proved for LDPE.

Low Density Nanocellular Polymers Based on PMMA Produced by Gas Dissolution Foaming: Fabrication and Cellular Structure Characterization.

This paper describes the processing conditions needed to produce low density nanocellular polymers based on polymethylmethacrylate (PMMA) with relative densities between 0.45 and 0.25, cell sizes between 200 and 250 nm and cell densities higher than 1014 cells/cm³. To produce these nanocellular polymers, the foaming parameters of the gas dissolution foaming technique using CO2 as blowing agent have been optimized. Taking into account previous works, the amount of CO2 uptake was maintained constant (31% by weight) for all the materials. Foaming parameters were modified between 40 °C and 110 °C for the foaming temperature and from 1 to 5 min for the foaming time. Foaming temperatures in the range of 80 to 100 °C and foaming times of 2 min allow for production of nanocellular polymers with relative densities as low as 0.25. Cellular structure has been studied in-depth to obtain the processing-cellular structure relationship. In addition, it has been proved that the glass transition temperature depends on the cellular structure. This effect is associated with a confinement of the polymer in the cell walls, and is one of the key reasons for the improved properties of nanocellular polymers.