RESUMO
In this work, a comparative analysis of the peripheral cell component (PCC) proteins of Listeria monocytogenes was carried out. The study was conducted on two set of samples consisting of bacteria treated with sub-lethal concentration of nisin and untreated bacteria as control. PCC proteins were extracted by Tris-Urea-EDTA treatment and then subjected to trypsin digestion and mass spectrometry analysis. The whole cell proteome was analyzed through label-free quantitative proteomics approach. Proteomic analysis was carried out using OrbiTrap Mass Spectrometer coupled to nanoflow liquid chromatography. The treatment with sub-lethal nisin concentration resulted in 62 up regulated and 97 down regulated proteins compared to untreated samples. Using PSORTb 3.0, 19 and 18 surface proteins were detected among the up regulated and down regulated proteins, respectively. Proteins related with increased biofilm formation by L.monocytogenes, such as moonlight proteins of the pyruvate dehydrogenase complex and flagellin-related proteins, were identified as up regulated surface proteins. Proteins associated with virulence of L.monocytogenes, including listeriolysin O, internalin B and actin assembly-inducing protein, were detected among the down regulated proteins. To confirm proteomics data, increased production of biofilm was experimentally confirmed in nisin-treated cells through crystal violet method. BIOLOGICAL SIGNIFICANCE: Proteosurfaceomics can be defined as the "omics" science applied to the proteins of the peripheral cell component (PCC). The surface proteins of Listeria monocytogenes, an important foodborne pathogen were investigated after treatment with nisin, a bacteriocin approved as a natural food preservative by regulatory agencies. Recent cases of nisin tolerance by Listeria spp. were documented, and deeper studies on the molecular process behind the bacterial survival may help in both understanding the development of tolerance process and comparing nisin effect with other antimicrobial compounds.
Assuntos
Listeria monocytogenes , Nisina , Antibacterianos , Proteínas de Membrana , Nisina/farmacologia , ProteômicaRESUMO
Air pollution has been identified as a major cause of environmental and human health damage. O3 is an oxidative pollutant that causes leaf symptoms in sensitive plants. This study aims to adjust a multilinear model for the monitoring of O3 in subtropical climatic conditions by associating O3 concentrations with measurements of morphological leaf traits in tobacco plants and different environmental variables. The plants were distributed into five areas (residential, urban or industrial) in the southern region of Brazil and exposed during 14 periods, of 14 days each, during the years of 2014 and 2015. The environmental variables and leaf traits during the exposure periods were described by mean, median, standard deviation and minimum and maximum values. Spearman correlation and multiple linear regression analyses were applied on data from exposure periods. Leaf injury index, leaf area, leaf dry mass, temperature, relative humidity, global solar radiation and accumulated rainfall were used in the regression analyses to select the best models for predicting O3 concentrations. Leaf injury characteristically caused by O3 was verified in all areas and periods of plant exposure. Higher values of leaf injury (24.5% and 27.7%) were registered in the 13th and 12th exposure periods during spring and in areas influenced by urban and industrial clutches. The VPD, temperature, global solar radiation and O3 were correlated to leaf injury. Environmental variables [leaf area, leaf dry mass, global solar radiation and accumulated rainfall] and primarily the VPD were fundamental to improve the adjustments done in the bioindicator model (R2â¯≥â¯0.73). Our research shows that biomonitoring employing the tobacco "Bel-W3" can be improved by measuring morphological leaf traits and meteorological parameters. Additionally, O3 fumigation experiment should be performed with biomonitoring as conducted in this study, which are useful in understanding the role of other environmental factors.