Contributing to Understand the Crosstalk between Brain and Periphery in Methylmercury Intoxication: Neurotoxicity and Extracellular Vesicles

Human exposure to methylmercury (MeHg) is currently high in regions such as the Amazon. Understanding the molecular changes associated with MeHg-induced neurotoxicity and the crosstalk with the periphery is essential to support early diagnoses. This work aimed to evaluate cellular and molecular changes associated with behavioral alterations in MeHg acute exposure and the possible changes in extracellular vesicles (EVs) number and S100β content. Adults male Wistar rats were orally treated with 5 mg/kg for four days. Behavioral performance, molecular and histological changes in the cerebellum, and plasma EVs were assessed. MeHg-intoxicated animals performed significantly worse in behavioral tests. MeHg increased the number of GFAP+ cells and GFAP and S100β mRNA expression in the cerebellum but no change in NeuN+ or IBA-1+ cells number was detected. The number of exosomes isolated from plasma were decreased by the metal. S100B mRNA was detected in circulating plasma EVs cargo in MeHg exposure. Though preliminary, our results suggest astrocytic reactivity is displaying a protective role once there was no neuronal death. Interestingly, the reduction in exosomes number could be a new mechanism associated with MeHg-induced neurotoxicity and plasma EVs could represent a source of future biomarkers in MeHg intoxication.     

Polyphenol Supplementation Reverses Age-Related Changes in Microglial Signaling Cascades

Microglial activity in the aging neuroimmune system is a central player in aging-related dysfunction. Aging alters microglial function via shifts in protein signaling cascades. These shifts can propagate neurodegenerative pathology. Therapeutics require a multifaceted approach to understand and address the stochastic nature of this process. Polyphenols offer one such means of rectifying age-related decline. Our group used mass spectrometry (MS) analysis to explicate the complex nature of these aging microglial pathways. In our first experiment, we compared primary microglia isolated from young and aged rats and identified 197 significantly differentially expressed proteins between these groups. Then, we performed bioinformatic analysis to explore differences in canonical signaling cascades related to microglial homeostasis and function with age. In a second experiment, we investigated changes to these pathways in aged animals after 30-day dietary supplementation with NT-020, which is a blend of polyphenols. We identified 144 differentially expressed proteins between the NT-020 group and the control diet group via MS analysis. Bioinformatic analysis predicted an NT-020 driven reversal in the upregulation of age-related canonical pathways that control inflammation, cellular metabolism, and proteostasis. Our results highlight salient aspects of microglial aging at the level of protein interactions and demonstrate a potential role of polyphenols as therapeutics for age-associated dysfunction.     

Novel Bacterial Production of Two Different Bioactive Forms of Human Stem-Cell Factor

Human stem-cell factor (hSCF) stimulates the survival, proliferation, and differentiation of hematopoietic cells by binding to the c-Kit receptor. Various applications of hSCF require the efficient and reliable production of hSCF. hSCF exists in three forms: as two membrane-spanning proteins hSCF248 and hSCF229 and truncated soluble N-terminal protein hSCF164. hSCF164 is known to be insoluble when expressed in Escherichia coli cytoplasm, requiring a complex refolding procedure. The activity of hSCF248 has never been studied. Here, we investigated novel production methods for recombinant hSCF164 and hSCF248 without the refolding process. To increase the solubility of hSCF164, maltose-binding protein (MBP) and protein disulfide isomerase b’a’ domain (PDIb’a’) tags were attached to the N-terminus of hSCF164. These fusion proteins were overexpressed in soluble form in the Origami 2(DE3) E. coli strain. These solubilization effects were enhanced at a low temperature. His-hSCF248, the poly-His tagged form of hSCF248, was expressed in a highly soluble form without a solubilization tag protein, which was unexpected because His-hSCF248 contains a transmembrane domain. hSCF164 was purified using affinity and ion-exchange chromatography, and His-hSCF248 was purified by ion-exchange and gel filtration chromatography. The purified proteins stimulated the proliferation of TF-1 cells. Interestingly, the EC₅₀ value of His-hSCF248 was 1 pg/mL, 100-fold lower than 9 ng/mL hSCF164. Additionally, His-hSCF248 decreased the doubling time, increased the proportion of S and G2/M stages in the cell cycle, and increased the c-Myc expression at a 1000-fold lower concentration than hSCF164. In conclusion, His-hSCF248 was expressed in a soluble form in E. coli and had stronger activity than hSCF164. The molecular chaperone, MBP, enabled the soluble overexpression of hSCF164.     

Importance of Reservoir Tributaries to Spawning of Migratory Fish in the Upper Paraná River

Regulation of rivers by dams transforms previously lotic reaches above the dam into lentic ones and limits or prevents longitudinal connectivity, which impairs access to suitable habitats for the reproduction of many migratory fish species. Frequently, unregulated tributaries can provide important habitat heterogeneity to a regulated river and may mitigate the influence of impoundments on the mainstem river. We evaluated the importance of tributaries to spawning of migratory fish species over three spawning seasons, by comparing several abiotic conditions and larval fish distributions in four rivers that are tributaries to an impounded reach of the Upper Paraná River, Brazil. Our study confirmed reproduction of at least 8 long‐distance migrators, likely nine, out of a total of 19 occurring in the Upper Paraná River. Total larval densities and percentage species composition differed among tributaries, but the differences were not consistent among spawning seasons and unexpectedly were not strongly related to annual differences in temperature and hydrology. We hypothesize that under present conditions, densities of larvae of migratory species may be better related to efficiency of fish passage facilities than to temperature and hydrology. Our study indicates that adult fish are finding suitable habitat for spawning in tributaries, fish eggs are developing into larvae, and larvae are finding suitable rearing space in lagoons adjacent to the tributaries. Our findings also suggest the need for establishment of protected areas in unregulated and lightly regulated tributaries to preserve essential spawning and nursery habitats. Copyright © 2014 John Wiley & Sons, Ltd.     

Organizational human factors as barriers to energy efficiency in electrical motors systems in industry

This paper presents a study accomplished in the State of Parana in Southern Brazil, aiming at verifying the correlation between organizational human factors (OHF) and the level of energy losses in organizations. The purpose is to subsidize the formularization of institutional policies in organizations to improve the energy efficiency in the productive sector. The research was carried out in ten industries of the following sectors: pulp and paper; food; wood and chemical products. The losses of electric energy were determined in motor systems with the aid of a mathematical model and the evaluation of 27 OHF identified in the literature review was made with the supervisors in the industries by means of a structurized questionnaire. Seven OHF had presented significant correlation with energy losses and six of them are inversely proportional to the losses, in accordance with linear regression analysis. The inversely proportional factors to the losses also with significant correlation are considered determinative OHF and constitute barriers for energy efficiency in organizations. These barriers are linked with the following organizational areas: management system; education of employees; strategical vision. The study recommends the implementation of the determinative OHF in organizations as a way to transpose the human barriers for energy efficiency.     

A theoretical study of the electronic structure and bonding of the monoclinic phase of

The electronic properties of the Mg₂NiH₄ monoclinic phase are calculated using a density functional approach calculation. The crystalline parameters and interatomic distances calculated are close to the experimental values within a 3% error. We also evaluate the density of states (DOS) and character of the chemical bonding for the hydrogen's located in their equilibrium positions. While the Ni–Mg interaction is dominant in the pure alloy, in the hydride the hydrogen atoms present a bonding much more developed with Ni than with Mg. The principal bonding interaction is Ni sp–H s. Moreover, a small bonding between Ni d_eg and H 1s is observed. Up the Fermi level, the Ni–H interaction is slightly antibonding. The Mg–Ni bonding interactions are weaker in the hydride phase when compared with the pure Mg₂Ni alloy. The present study is potentially useful because the alloys Mg–Ni are good materials for hydrogen storage.     

Adherence kinetics, resistance to benzalkonium chloride and microscopic analysis of mixed biofilms formed by Listeria monocytogenes and Pseudomonas putida

Comparison between the resistance to BAC and the microscopic structure between mixed-species biofilms formed by different strains of Listeria monocytogenes and Pseudomonas putida CECT 845 under different scenarios and that obtained by the corresponding monospecies L. monocytogenes biofilm was carried out. The association of P. putida with L. monocytogenes quickens biofilm formation and increases significantly (p < 0.05) the BAC-resistance of the biofilm after 4 days of incubation at 25 °C respecting to that formed by monospecies biofilms. According with the adherence profiles of P. putida, two different patterns of association between both species (A and B) were identified, being type A pattern found in the mixed biofilms much more resistant to BAC. After 11 days of incubation, a destructuration of mixed biofilms occurred in all experimental assays, being in 2 out of 5 experimental cases (4032 and BAC-adapted 5873 on polypropylene) accompanied by a sharp decrease in the number of adhered cells. Microscopic analyses demonstrated that complex three-dimensional microscopic structure showed the highest resistance to BAC (4032-SS). Obtained results clearly highlight that to improve disinfection protocols for assuring food safety, it is necessary to mimick those bacterial association that occur in nature.     

Survival of Listeria innocua in Minas Traditional Serro cheese during ripening

Cheese produced with raw milk can be a risk to consumer health. It is known that lactic acid bacteria present in raw milk and in natural starters can produce antimicrobial compounds against some foodborne bacteria. This work aimed to evaluate the survival of Listeria innocua in Minas Traditional Serro cheese during cheese ripening. The cheeses were inoculated with 10¹, 10² or 10³ CFU mL⁻¹ of the bacterium and were analyzed for 60 days of ripening at 30 °C. It was observed that the time and the dose of bacteria inoculated affected (p < 0.05) the survival of L. innocua. Even when the lowest dose was inoculated, at the end of the 60 days, approximately 10² CFU mL⁻¹ of L. innocua was detected in the cheese. The lactic acid bacteria present in the milk and in the natural starter were not sufficient to guarantee the absence of L. innocua in Minas Traditional Serro cheese even after 60 days of storage, as is required by Brazilian legislation.     

Evaluation of a bacteriocin-producing strain of Pediococcus acidilactici as a biopreservative for “Alheira”, a fermented meat sausage

This study was conducted to evaluate the ability of Pediococcus acidilactici HA-6111-2, a PA-1 bacteriocin-producing lactic acid bacterium (LAB), isolated from “Alheira” to inhibit a cocktail of Listeria innocua strains during production and shelf-life of these products. The bacteriocinogenic culture reduced the Listeria population below the detection limit (1.5log CFU/g) and had no effect on the growth of the natural LAB flora or on the pH. Pathogenic organisms were not detected in any sample. The presence of some virulence factors and antibiotic resistances of the strain to be used as a bioprotective culture were investigated. P. acidilactici HA-6111-2 did not produce any of the biogenic amines tested; no formation of biofilms was observed; more l(+)lactic acid was produced than its isomer d(?); no gelatinase, DNase or lipase activity was recorded; no structural genes for the haemolysin, enterococcal surface protein, hydrolytic compounds, aggregation protein and cell-wall adhesins were detected, no significant antibiotic resistances were found. P. acidilactici HA-6111-2 appears to have potential as a bioprotective culture during “Alheira” fermentation. Moreover, a trained panel considered the protected product to be sensorially acceptable.     

Non-catalytic production of fatty acid ethyl esters from soybean oil with supercritical ethanol in a two-step process using a microtube reactor

This work reports the production of fatty acid ethyl esters (FAEE) from the transesterification of soybean oil in supercritical ethanol in a continuous catalyst-free process using different reactor configurations. Experiments were performed in a microtube reactor with experimental simulation of two reactors operated in series and a reactor with recycle, both configurations at a constant temperature of 573 K, pressure of 20 MPa and oil to ethanol mass ratio of 1:1. Results show that the configurations studied with intermediate separation of glycerol afford higher conversions of vegetable oil to its fatty acid ethyl ester derivatives when compared to the one-step reaction, with relatively low decomposition of fatty acids (<3.0 wt%).