PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.

Pseudomonas aeruginosa, which is a clinically important representative of Pseudomonas spp., has been recognized as causative agent of severe nosocomial infections worldwide. An increase in antibiotic resistance of P. aeruginosa clinical strains could be attributed to their capacity to acquire resistance through mobile genetic elements such as mobile integrons that are present in one-half of multidrug-resistant P. aeruginosa strains. Mobile class 1 integrons are recognized as genetic elements involved in the rapid dissemination of multiple genes encoding for antibiotic resistance. The LexA protein is a major repressor of integrase transcription, but differences in transcription regulation among bacterial species have also been noted. In this study, the promoter activity of class 1 integron integrase gene (intI1) and its variant lacking the LexA binding site in Pseudomonas putida WCS358 wild type, ΔrpoS and ΔpsrA was analysed. The results show that the activity of the intI1 gene promoter decreased in the rpoS and psrA mutants in the stationary phase of growth compared to the wild type, which indicates the role of RpoS and PsrA proteins in the positive regulation of integrase transcription. Additionally, it was determined that the activity of the lexA gene promoter decreased in ΔrpoS and ΔpsrA, and thus, we propose that PsrA indirectly regulates the intI1 gene promoter activity through regulation of lexA gene expression in co-operation with some additional regulators. In this study, intI1 gene expression was shown to be controlled by two major stress response (SOS and RpoS) regulons, which indicates that integrase has evolved to use both systems to sense the cell status.

Polyphenol bioaccessibility and antioxidant properties of in vitro digested spray-dried thermally-treated skimmed goat milk enriched with pollen.

The aim of research was to determine polyphenols bioaccessibility and antioxidant properties of thermally-treated skimmed goat milk enriched with sunflower bee-collected pollen through in vitro digestion. HPLC analysis confirmed that pollen-enriched milk contained flavonols as the main phenolic fraction (80.7-76.2%) followed by phenolic acids (14.2-17.4%). Among individual compounds quercetin-3-O-glucoside (155.1-197.2 µg/L) and p-coumaric acid (29.5-30.7 µg/L) were the main quantified flavonols and phenolic acids, respectively. After digestion of milk/pollen sample, total polyphenols recovery was 30.71% with higher phenolic acids recovery (40.1%) compared to flavonols (28.3%) indicating strong interactions between caprine milk casein micelles and pollen polyphenols. Applied antioxidant assays (phosphomolybdenum, ABTS•+scavenging activity and ferrous-ion-chelating capacity) have confirmed complexity of prepared product- it had high ability to quench ABTS•+ radicals and to form chelating complexes with Fe2+ ions. Digestion provoked 20% reduction in total antioxidant capacity compared to the initial sample. TTSG milk/pollen powder could be good functional ingredient.

In vitro assessment of pesticide residues bioaccessibility in conventionally grown blueberries as affected by complex food matrix.

The aim of this study was to investigate the bioaccessibility of pesticide residues in blueberries (commercial and sample from controlled field trial) from Serbia, involving the presence of a complex food matrix and to assess the potential risk to human health. The presence of nine active substances (azoxystrobin, boscalid, fludioxonil, cyprodinil, pyrimethanil, pyridaben, pyriproxyfen, acetamiprid and thiametoxam) in initial blueberry samples was determined in concentration range from 5.15 µg/kg for thiametoxam to 187 µg/kg for azoxystrobin. Clothianidin, metabolite of thiametoxam, was not detected in any blueberry sample. However, after in vitro digestion, the content of initially detected pesticides residues was significantly decreased or it was below limit of quantification resulting in the total bioaccessibility of about 15%. Azoxystrobin, pyrimethanil and fludioxonil was quantified in digestive juice at concentrations which were about 81%, 37% and 10% less than the inital concentration, respectively. The presence of food matrix during digestion of blueberries even more severely reduced concentration of pesticide residues (total bioaccessibility was about 7%) compared to digestion without the food matrix. Only azoxystrobin was quantified after digestion with food matrix in concentration of 27 µg/kg in sample from controlled field trial and detected in two commercial samples but below the limit of quantification. Furthermore, chronic risk assessment indicated that risk is acceptable for the health of different human subpopulation groups. The current study on pesticides residues, most commonly applied on blueberries, provides for the first time an insight into their bioaccessibility under conditions that mimic physiological environment of human digestive tract.

In vitro digestion of meat- and cereal-based food matrix enriched with grape extracts: How are polyphenol composition, bioaccessibility and antioxidant activity affected?

The aim of this study was to evaluate the effect of enriching a complex food matrix (FM) with grape extracts on polyphenol content, composition, bioaccessibility and antioxidant activity during digestion. The grape extracts and FM were separately tested under the same conditions as controls. The FM by itself contains a significant amount of phenolic acids and flavonols, influencing the final recovery of polyphenols from grape extracts. The FM significantly increased the total recovery of polyphenols after digestion of grape seed extracts compared to those digested without the FM; however, a low recovery of proantocyanidins and total flavonoids was observed. Digestive fluids and FM compounds significantly increased the total polyphenol content of grape digests and significantly contributed to their ABTS+ scavenging activity and ferrous-ion-chelating capacity. The present study suggested that enrichment of meat- and cereal-based products with grape polyphenol extracts could be a good strategy to formulate a healthier diet.