•The volatile exometabolome of three strains of S. aureus was studied.•The volatile exometabolome each strain comprised in average 240 metabolites.•Hierarchical clustering analysis allowed the ...strains metabotyping.•Enterotoxic and non-enterotoxic strains exhibited distinct metabolomic profiles.
Staphylococcal food poisoning is a disease that originates significant health and economic losses and is caused by Staphylococcus aureus strains able to produce enterotoxins. The aim of this work is to go further on the study of the volatile exometabolome of S. aureus using an advanced gas chromatographic technique. Enterotoxic and non-enterotoxic strains were assessed. The volatile exometabolome profile comprised 240 volatiles belonging to ten chemical families. This volatiles were mainly by-products of branched-chain amino acids and methionine degradation, pyruvate metabolism, diacetyl pathway, oxidative stress and carotenoid cleavage. Metabolites released by the first two pathways were produced in higher contents by the enterotoxic strains. This study add further insights to S. aureus volatile exometabolome, and also shows that by applying it, it is possible to distinguish strains of S. aureus by the number of produced enterotoxins, which is especially important from the food safety point of view.
To investigate the subjective experience of adult cancer patients undergoing external radiotherapy and provide evidence for better practices in radiotherapy services.
A systematic review was ...performed according to the PRISMA Statement Guidelines. Qualitative and mixed studies were identified through five electronic databases (CINAHL, PsychINFO, Medline, Scopus and Web of Science), between March and April 2020, using defined criteria. Methodological quality assessment was conducted, and the data integrated into a thematic synthesis.
Of the 886 studies identified, 13 met our inclusion criteria. Patients experiences were described into four main themes: (1) Time, (2) Physical Environment, (3) Treatment Concerns and (4) Radiotherapy Team. Time refers to waiting time and treatment time; Physical Environment states temperature in the treatment room and equipment; Treatment Concerns included side effects, daily activities, positioning and immobilization and treatment preparation (e.g., bladder filling); radiotherapy team comprised patients support; response to patients needs and recognized team. The generating analytical phase of thematic analysis allowed us to recognize that the effectiveness of the radiotherapy team operates as a facilitator of the patients' experience. In contrast, time in the waiting room, the treatment preparation when is required, and the positioning and immobilization, specifically for the neck and head patients, act as barriers.
Four distinct themes were identified to be positively and negatively associated with patients' experience during external radiotherapy. The evidence provides valuable recommendations to improved radiotherapy services organization, as well as to the delivery of more patient-centred care adjusted to the concerns and needs of patients.
•More than 50% of cancer patients are treated using radiation treatment.•An overview of the evidence considering the experience-related themes is required in radiotherapy care.•Understanding the facilitators and barriers of cancer patients' experiences provides enables radiotherapy best practices.
A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-ToFMS) was developed ...for the simultaneous screening of microbial and mite contamination level in cereals and coffee beans. The proposed approach emerges as a powerful tool for the rapid assessment of the microbial contamination level (ca. 70 min versus ca. 72 to 120 h for bacteria and fungi, respectively, using conventional plate counts), and mite contamination (ca. 70 min versus ca. 24 h). A full-factorial design was performed for optimization of the SPME experimental parameters. The methodology was applied to three types of rice (rough, brown, and white rice), oat, wheat, and green and roasted coffee beans. Simultaneously, microbiological analysis of the samples (total aerobic microorganisms, moulds, and yeasts) was performed by conventional plate counts. A set of 54 volatile markers was selected among all the compounds detected by GC×GC-ToFMS. Principal Component Analysis (PCA) was applied in order to establish a relationship between potential volatile markers and the level of microbial contamination. Methylbenzene, 3-octanone, 2-nonanone, 2-methyl-3-pentanol, 1-octen-3-ol, and 2-hexanone were associated to samples with higher microbial contamination level, especially in rough rice. Moreover, oat exhibited a high GC peak area of 2-hydroxy-6-methylbenzaldehyde, a sexual and alarm pheromone for adult mites, which in the other matrices appeared as a trace component. The number of mites detected in oat grains was correlated to the GC peak area of the pheromone. The HS-SPME/GC×GC-ToFMS methodology can be regarded as the basis for the development of a rapid and versatile method that can be applied in industry to the simultaneous assessment the level of microbiological contamination and for detection of mites in cereals grains and coffee beans.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Closely spaced promoters in tandem formation are abundant in bacteria. We investigated the evolutionary conservation, biological functions, and the RNA and single-cell protein expression of genes ...regulated by tandem promoters in E. coli. We also studied the sequence (distance between transcription start sites 'dTSS', pause sequences, and distances from oriC) and potential influence of the input transcription factors of these promoters. From this, we propose an analytical model of gene expression based on measured expression dynamics, where RNAP-promoter occupancy times and dTSS are the key regulators of transcription interference due to TSS occlusion by RNAP at one of the promoters (when dTSS ≤ 35 bp) and RNAP occupancy of the downstream promoter (when dTSS > 35 bp). Occlusion and downstream promoter occupancy are modeled as linear functions of occupancy time, while the influence of dTSS is implemented by a continuous step function, fit to in vivo data on mean single-cell protein numbers of 30 natural genes controlled by tandem promoters. The best-fitting step is at 35 bp, matching the length of DNA occupied by RNAP in the open complex formation. This model accurately predicts the squared coefficient of variation and skewness of the natural single-cell protein numbers as a function of dTSS. Additional predictions suggest that promoters in tandem formation can cover a wide range of transcription dynamics within realistic intervals of parameter values. By accurately capturing the dynamics of these promoters, this model can be helpful to predict the dynamics of new promoters and contribute to the expansion of the repertoire of expression dynamics available to synthetic genetic constructs.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Tributyltin (TBT) is a biocide extremely toxic to a wide range of organisms, which has been used for decades in antifouling paints. Despite its global ban in 2008, TBT is still a problem of great ...concern due to the high levels trapped in sediments. Aeromonas molluscorum Av27 is a TBT degrading bacterium that was isolated from an estuarine system. We investigated the ability and the role of this bacterium on TBT degradation in this estuarine system, using a microcosm approach in order to mimic environmental conditions. The experiment was established and followed for 150 days. Simultaneously, changes in the indigenous bacterial community structure were also investigated. The results revealed a maximum TBT degradation rate of 28% accompanied by the detection of the degradation products over time. Additionally, it was observed that TBT degradation was significantly enhanced by the presence of Av27. In addition a significantly higher TBT degradation occurred when the concentration of Av27 was higher. TBT degradation affected the bacterial community composition as revealed by the changes in the prevalence of Proteobacteria subdivisions, namely the increase of Deltaproteobacteria and the onset of Epsilonproteobacteria. However, the addition of Av27 strain did not affect the dominant phylotypes. Total bacterial number, bacterial biomass productivity, 16S rRNA gene and denaturing gradient gel electrophoresis (DGGE) analyses also indicated alterations on the bacterial community structure over time, with bacteria non-tolerant to pollutants increasing their representativeness, as, for instance, the increase of the number of Alphaproteobacteria clones from 6% in the beginning to 12% at the end of the experiment. The work herein presented confirms the potential of Av27 strain to be used in the decontamination of TBT-polluted environments.
•A microcosm approach was used to evaluate TBT degradation by A. molluscorum Av27.•Av27 significantly enhanced TBT degradation in estuarine sediment over time.•Higher concentration of Av27 induces a significantly higher TBT degradation rate.•Av27 induced alterations in the diversity and composition of the bacterial community.•Induced alterations did not affect the dominant phylotypes.
Abstract
Bacteria from the surface microlayer (bacterioneuston) and underlying waters (bacterioplankton) were isolated upon exposure to UV-B radiation, and their individual UV sensitivity in terms of ...CFU numbers, activity (leucine and thymidine incorporation), sole-carbon source use profiles, repair potential (light-dependent and independent), and photoadaptation potential, under different physiological conditions, was compared. Colony counts were 11.5–16.2% more reduced by UV-B exposure in bacterioplankton isolates (P < 0.05). Inhibition of leucine incorporation in bacterioneuston isolates was 10.9–11.5% higher than in bacterioplankton (P < 0.05). These effects were accompanied by a shift in sole-carbon source use profiles, assessed with Biolog® EcoPlates, with a reduction in consumption of amines and amino acids and increased use of polymers, particularly in bacterioneuston isolates. Recovery under starvation was generally enhanced compared with nourished conditions, especially in bacterioneuston isolates. Overall, only insignificant increases in the induction of antibiotic resistant mutant phenotypes (RifR and NalR) were observed. In general, a potential for photoadaptation could not be detected among the tested isolates. These results indicate that UV effects on bacteria are influenced by their physiological condition and are accompanied by a shift in metabolic profiles, more significant in bacterioneuston isolates, suggesting the presence of bacterial strains adapted to high UV levels in the SML.
Staphylococcus aureus is responsible for a large spectrum of diseases, including staphylococcal food poisoning (SFP), due to its ability to produce enterotoxins. To prevent the development of SFP, ...effective food preservation methods are needed. High pressure processing (HPP) uses pressures, between 100 and 600 MPa, to inactivate pathogenic and spoilage microorganisms. In this study, HPP treatments (450 MPa and 600 MPa, both for 15 and 30 min, at 20 °C) were applied in three different strains of S. aureus in the stationary growth phase and reduction effectiveness was assessed. A non-enterotoxic strain ATCC 6538 and two enterotoxic strains 2153 MA (with enterotoxin A) and 2065 MA (with enterotoxin A, G, I) were used. It was verified that the non-enterotoxic strain was the most resistant to HPP, not being completely inactivated within 30 min at 600 MPa. Additionally, it was demonstrated that HPP had no effect on virulence factors (enterotoxins, β-hemolysin, lipase, lecithinase, coagulase, thermonuclease, catalase), and also in mannitol fermentation capacity and methicillin susceptibility. HPP treatments also proved to be less effective in the strain with higher carotenoids content (non-enterotoxic strain). The results of this study show that S. aureus HPP barotolerance might be related not only to the presence of enterotoxins but also to carotenoids level, although these two factors may not be the only mechanisms responsible for the distinct sensitivity/resistance to HPP shown by different strains of S. aureus.
•Staphylococcus aureus strains were submitted to high pressure processing (HPP) treatments.•The effect on cell viability and the impact on virulence factors was assessed.•Strains with enterotoxins and low carotenoids content were less baroresistant.•Enterotoxins and other virulence factors/characteristics under study were not affected by HPP.
Abstract
Intrinsic and acquired resistance to chemotherapy is the fundamental reason for treatment failure for many cancer patients. The identification of molecular mechanisms involved in drug ...resistance or sensitization is imperative. Here we report that tribbles homologue 2 (TRIB2) ablates forkhead box O activation and disrupts the p53/MDM2 regulatory axis, conferring resistance to various chemotherapeutics. TRIB2 suppression is exerted via direct interaction with AKT a key signalling protein in cell proliferation, survival and metabolism pathways. Ectopic or intrinsic high expression of TRIB2 induces drug resistance by promoting phospho-AKT (at Ser473) via its COP1 domain. TRIB2 expression is significantly increased in tumour tissues from patients correlating with an increased phosphorylation of AKT, FOXO3a, MDM2 and an impaired therapeutic response. This culminates in an extremely poor clinical outcome. Our study reveals a novel regulatory mechanism underlying drug resistance and suggests that TRIB2 functions as a regulatory component of the PI3K network, activating AKT in cancer cells.
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