A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

Background: Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. Results: An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions: Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far

Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release

Measure of Customer Satisfaction in the Residential Electricity Distribution Service Using Structural Equation Modeling

The main objective of this study is to apply structural equation modeling with partial least squares, and based on covariance, to assess the satisfaction of residential electricity consumers. The methodology used compares the results of both structural equation models to indicate the model that best fits the problem of measuring the satisfaction of residential consumers with electricity concessionaires and licensees. The sample used in the survey contained questionnaire responses from 86,175 individuals considering the period from 2014 to 2018. The constructs evaluated were satisfaction, quality, value, loyalty, and trust. A confidence interval analysis shows that all weights are significant, demonstrating the importance of all the indicators that represent the constructs. The trust, quality, and value constructs can explain 74.4% of the satisfaction construct variability, so this relationship’s explanatory capacity is considered substantial. Finally, the evaluation of the performance of the service provided by the electric energy concessionaires/licensees, measured by customer satisfaction, allows for the continuous improvement of services, and meeting, even if minimally, the expectations of its consumers

Chemical Composition and Ruminal Degradability of Spineless Cactus Grown in Northeastern Brazil

A study was conducted to determine chemical composition and ruminal nutrient degradability of eight spineless cactus cultivars grown in northeastern Brazil. Results showed that neutral detergent fiber was similar for all cultivars and averaged 249 g kg-1 +/- 7.3 SEM. Acid detergent fiber ranged between 148 g kg-1 and 207 g kg-1 with some significant differences between cultivars. Starch and water-soluble carbohydrates were similar for all cultivars and averaged 198 g kg-1 +/- 6.3 SEM and 155 g kg-1 +/- 9.0 SEM, respectively. Protein content was less than 50 g kg-1 with some significant differences between cultivars. Calcium was the mineral with the highest concentration followed by potassium and magnesium with no differences between cultivars. Effective ruminal degradability of dry matter and neutral detergent fiber were unaffected by cultivar and averaged 701 g kg-1 +/- 8.4 SEM and 503 g kg-1 +/- 5.8 SEM, respectively. It was concluded that cultivars had little impact on chemical composition and ruminal degradability of spineless cactus. Based on chemical composition and in situ ruminal degradability, spineless cactus can be considered an excellent source of fermentable carbohydrates for grazing and nongrazing ruminants. Because of its high carbohydrate quality, spineless cactus can be used an emergency feed or as part of a complete diet providing that the diet contains an adequate amount of degradable protein. The Rangeland Ecology & Management archives are made available by the Society for Range Management and the University of Arizona Libraries. Contact [email protected] for further information.Migrated from OJS platform August 202

Exposure to an extremely low-frequency electromagnetic field only slightly modifies the proteome of Chromobacterium violaceum ATCC 12472

Several studies of the physiological responses of different organisms exposed to extremely low-frequency electromagnetic fields (ELF-EMF) have been described. In this work, we report the minimal effects of in situ exposure to ELF-EMF on the global protein expression of Chromobacterium violaceum using a gel-based proteomic approach. The protein expression profile was only slightly altered, with five differentially expressed proteins detected in the exposed cultures; two of these proteins (DNA-binding stress protein, Dps, and alcohol dehydrogenase) were identified by MS/MS. The enhanced expression of Dps possibly helped to prevent physical damage to DNA. Although small, the changes in protein expression observed here were probably beneficial in helping the bacteria to adapt to the stress generated by the electromagnetic field

Determinação de Cu, Fe, Mn, Zn e do teor de proteína total em amostras de trigo e soja após procedimento de extração sequencial

A sequential extraction procedure was applied to wheat and soybean seed samples. The total protein content (determined by two distinct methods: Bradford and bicinchoninic acid-BCA) and distribution of Cu, Fe, Mn and Zn in each fraction was determined. The sequential extraction employed four different solutions: water, 0.5 mol L-1 NaCl, ethanol/water (70:30 v v-1) and 0.5 mol L-1 NaOH. For both samples, the highest concentration of metals was observed in those extracts associated with globulin-type proteins using NaCl solution. Regarding protein content, higher levels were obtained using the BCA method