Enhanced removal of the endocrine disruptor compound Bisphenol A by adsorption onto green-carbon materials. Effect of real effluents on the adsorption process.

The high exposure to the endocrine disrupting compounds (EDC) in water represents a relevant issue for the health of living beings. The xenoestrogen Bisphenol A (BPA), a suspected EDC, is an industrial additive broadly used for manufacturing polycarbonate and epoxy resins. Due to its harmful effect in humans and the aquatic environment, an efficient method to remove BPA from wastewater is urgently required. The present work aims to study the adsorption of BPA from aqueous solutions onto carbonaceous materials, e.g., a synthesized carbon xerogel (RFX), a chemical-activated carbon from Kraft lignin (KLP) and a commercial activated carbon (F400) for comparative purposes. Batch kinetic and adsorption tests of BPA in ultrapure water were accomplished, finding higher adsorption capacities of BPA onto both F400 activated carbon (qsat = 407 mg g-1) and the biochar KLP (qsat = 220 mg g-1), versus to that obtained for the xerogel (qsat = 78 mg g-1). Furthermore, kinetic experiments revealed faster kinetic adsorption for RFX and KLP materials, achieving the equilibrium time within 24 h, attributed to their more-opened porous structure. Pseudo-first order, pseudo-second order, Elovich, intra-particle diffusion and film diffusion models were used to fit the experimental data. Thus, the BPA adsorption isotherms were analysed by Langmuir, Freundlich, Sips, Redlich-Peterson and Dual-site Langmuir (DLS) isotherm models.In addition, the influence of different aqueous matrices, such as a hospital wastewater, a wastewater treatment plant (WWTP) effluent and a river water, on BPA removal efficiency has been explored. These adsorption tests revealed a clear competitive effect between the target compound (BPA) and the natural organic matter content (NOM) present in the matrices for the active sites, resulting in a high decreasing of BPA adsorption removal.

A novel influenza A virus mitochondrial protein that induces cell death.

While searching for alternative reading-frame peptides encoded by influenza A virus that are recognized by CD8+ T cells, we found an abundant immunogenic peptide encoded by the +1 reading frame of PB1. This peptide derives from a novel conserved 87-residue protein, PB1-F2, which has several unusual features compared with other influenza gene products in addition to its mode of translation. These include its absence from some animal (particularly swine) influenza virus isolates, variable expression in individual infected cells, rapid proteasome-dependent degradation and mitochondrial localization. Exposure of cells to a synthetic version of PB1-F2 induces apoptosis, and influenza viruses with targeted mutations that interfere with PB1-F2 expression induce less extensive apoptosis in human monocytic cells than those with intact PB1-F2. We propose that PB1-F2 functions to kill host immune cells responding to influenza virus infection.

New insights from modelling and estimation of mass transfer parameters in fixed-bed adsorption of Bisphenol A onto carbon materials.

The removal of Bisphenol A, 2,2-bis (4-hydroxyphenyl) propane (BPA) in fixed-bed columns was investigated by breakthrough adsorption tests at different operation conditions and further prediction by a mathematical model to describe the adsorption-diffusion process onto two synthesized carbon porous materials. In this study, a xerogel (RFX) prepared by an optimized conventional sol-gel method and a lignin-based activated carbon (KLP) obtained via chemical activation were used in batch and fixed-bed adsorption experiments. The materials were fully characterized and their adsorptive properties were compared to those obtained with a commercial activated carbon (F400). RFX and KLP materials reached the equilibrium adsorption in only 24 h, whereas F400 activated carbon required 48 h. In addition, F400 and KLP adsorbents showed higher equilibrium adsorption capacity values (qe = 0.40 and 0.22 kg/kg, for F400 and KLP, respectively) than that obtained for the xerogel (qe = 0.08 kg/kg). Both synthesized carbon-adsorbents were studied in fixed-bed adsorption tests, exploring the effect of the operation conditions, e.g., initial BPA concentration (0.005-0.04 kg/m3), weight of adsorbent (0.01-0.05 g) and volumetric flow rate (0.2 to 1.0 mL/min), on the adsorption performance of the column. All the tested adsorption columns reached the equilibrium in a very short time, due to the efficient dimensionless of the bed. Additionally, the regeneration of the exhausted adsorbent was studied, achieving the total reuse of the solids after three consecutive cycles using methanol as regeneration agent. Finally, a mathematical model based on mass conservation equations was proposed, allowing to efficiently fit the experimental BPA breakthrough curves and estimate the external and adsorbed-phase mass transfer coefficients with a high accuracy.

Plasmodium yoelii: the role of the individual epidermal growth factor-like domains of the merozoite surface protein-1 in protection from malaria.

The merozoite surface protein-1 (MSP-1) is a leading candidate for a vaccine targeted at the erythrocytic stages of plasmodial parasite development. Recently, there has been increasing interest in this polypeptide, particularly in the carboxyl-terminal EGF-like domains. We have previously shown that this region from Plasmodium yoelii, when expressed in native configuration, could immunize mice against an otherwise lethal challenge infection. In this model system, protection appears to be predominantly mediated by antibodies. In all rodent immunization studies to date, however, the immunogen has contained both of the postulated EGF-like domains. We report here on the efficacy of immunization with the individual EGF-like domains from P. yoelii in elicitation of a protective host response. Although all animals developed some level of antibody in response to the various immunogens, only those animals immunized with both EGF-like domains produced antibodies which could recognize the native MSP-1 molecule. Antibodies generated against the individual EGF-like domains did cross-react with the double EGF-like domain structure, suggesting that the immunogens had retained elements of native configuration. In addition, only those animals which generated antibodies capable of recognizing native MSP-1 showed any level of protection from challenge infection. These results suggest that determinants unique to the double EGF-like domain structure may be necessary for the generation of antibodies specific for the native configuration of MSP-1 and that these antibodies may play a significant role in protection.

Phenotypes associated with malignant hyperthermia susceptibility in swine genotyped as homozygous or heterozygous for the ryanodine receptor mutation.

We have examined the phenotypic expression of several parameters associated with malignant hyperthermia (MH) susceptibility in three groups (homozygous normal, homozygous abnormal and heterozygous) of Yorkshire/Duroc swine genotyped by a mutation in the ryanodine receptor. Subgroups of homozygous abnormals were classified further by the appearance or absence of muscle rigidity on prolonged in vivo challenge with halothane and suxamethonium. Four swine heterozygous for the proposed MH mutation were indistinguishable from five homozygous normal swine in temperature, heart rate, lactate concentrations, base excess and pH determined during the prolonged halothane and suxamethonium challenge. Resting creatine kinase concentrations, the in vivo barnyard challenge, the in vitro contracture response of skeletal muscle to 3% halothane and the threshold for Ca(2+)-induced Ca2+ release were also similar for subgroups of homozygous normals and heterozygotes. Therefore, inheritance of only one allele carrying the defect in the ryanodine receptor does not significantly alter phenotypes associated with MH susceptibility in this strain of swine. As four swine homozygous for the proposed MH defect did not exhibit rigidity and three of these had no other signs of MH on prolonged halothane and suxamethonium challenge, we conclude that the reported mutation in the ryanodine receptor may be necessary, but is not sufficient, for consistently eliciting the malignant hyperthermia syndrome. These findings suggest that a modulator of the syndrome may explain variability within individuals in human MH.

Comparison of humoral immune responses elicited by DNA and protein vaccines based on merozoite surface protein-1 from Plasmodium yoelii, a rodent malaria parasite.

Immunization with DNA vaccines encoding relevant Ags can induce not only cell-mediated immune response but also humoral immune responses against pathogenic microorganisms in several animal models. Our previous results demonstrated that, when the C terminus (PyC2) of Plasmodium yoelii merozoite surface protein-1 (MSP-1), a leading vaccine candidate against erythrocytic stages of malaria, was expressed as a fusion protein (GST-PyC2) with glutathione S-transferase (GST), it elicited Ab-mediated protective immune responses in BALB/c mice. In our present study, we wished to examine the humoral responses to a DNA vaccine (V3) encoding GST-PyC2. The GST-PyC2 expressed in V3-transfected Cos 7 cells was recognized by a protective monoclonal Ab to PyC2 (mAb302), although the secreted product had undergone N-linked glycosylation. When BALB/c mice were immunized with V3 plasmid, anti-PyC2 Abs were successfully induced. These Abs immunoprecipitated native PyMSP-1 protein and competed with mAb302 for binding to its epitope at a level similar to those elicited by GST-PyC2 protein immunization. However, these Abs had significantly lower titers and avidities, and different isotype profiles and protective capacities against a lethal erythrocytic stage challenge, than those resulting from immunization with GST-PyC2 protein. Most surprising was the finding that, in contrast to protein immunization, there was no significant increase in the avidity of either GST-specific or PyC2-specific IgG Abs during the course of DNA immunization. This suggests that there may be little or no affinity maturation of specific Abs during DNA immunization in this system.

A common temperature-sensitive allelic form of human tyrosinase is retained in the endoplasmic reticulum at the nonpermissive temperature.

Oculocutaneous albinism type 1TS is caused by mutations that render the melanocyte-specific enzyme tyrosinase temperature-sensitive (ts); the enzyme is inactive in cells grown at 37 degrees C but displays full activity in cells grown at 31 degrees C. To distinguish whether the ts phenotype of the common R402Q variant of human tyrosinase is due to altered enzymatic activity or to misfolding and a defect in intracellular trafficking, we analyzed its localization and processing in transiently transfected HeLa cells. R402Q tyrosinase accumulates in the endoplasmic reticulum (ER) at 37 degrees C but exits the ER and accumulates in endosomal structures in cells grown at 31 degrees C. The inability of the R402Q variant to exit the ER is confirmed by the failure to acquire endoglycosidase H resistance at 37 degrees C and cannot be accounted for solely by enhanced proteasome-mediated degradation. ER retention at 37 degrees C is mediated by the lumenal domain of R402Q tyrosinase, is not dependent on tethering to the membrane, and is irreversible. Finally, a wild-type allelic form of tyrosinase is partially ts in transiently transfected HeLa cells. The data show that human tyrosinase expressed in non-melanogenic cells folds and exits the ER inefficiently and that R402Q tyrosinase exaggerates this defect, resulting in a failure to exit the ER at physiologic temperatures.

A cytoplasmic sequence in human tyrosinase defines a second class of di-leucine-based sorting signals for late endosomal and lysosomal delivery.

Distinct cytoplasmic sorting signals target integral membrane proteins to late endosomal compartments, but it is not known whether different signals direct targeting by different pathways. The availability of multiple pathways may permit some cell types to divert proteins to specialized compartments, such as the melanosome of pigmented cells. To address this issue, we characterized sorting determinants of tyrosinase, a tissue-specific resident protein of the melanosome. The cytoplasmic domain of tyrosinase was both necessary and sufficient for internalization and steady state localization to late endosomes and lysosomes in HeLa cells. Mutagenesis of two leucine residues within a conventional di-leucine motif ablated late endosomal localization. However, the properties of this di-leucine-based signal were distinguished from that of CD3gamma by overexpression studies; overexpression of the tyrosinase signal, but not the well characterized CD3gamma signal, induced a 4-fold enlargement of late endosomes and lysosomes and interfered with endosomal sorting mediated by both tyrosine- and other di-leucine-based signals. These properties suggest that the tyrosinase and CD3gamma di-leucine signals are distinctly recognized and sorted by distinct pathways to late endosomes in non-pigmented cells. We speculate that melanocytic cells utilize the second pathway to divert proteins to the melanosome.