► An UHPLC–MS/MS method was applied to quantify the phenolic compounds of olive oils. ► Analysis of phenolic compounds in fourteen oil samples belonging to different varieties. ► Significant ...differences of phenolic content between some varieties of oils were found.
In this study a simultaneous determination of several classes of polyphenolic compounds (benzoic acid derivates, cinnamic acid derivates, phenyl ethyl alcohols, flavones and other phenolic acids) in feed oils has been carried out by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS). Solid-phase extraction (SPE) with Diol and C18 cartridges were compared in terms of recovery and number of extracted compounds, obtaining better results when Diol cartridges were used. The analytical procedure was validated, obtaining recoveries ranging from 70% (vanillic acid) to 111% (gallic acid) with repeatability values (expressed as relative standard deviations, RSDs) lower than 20% at two concentration levels (500 and 1000μg/kg). Limits of quantification (LOQs) were always equal or lower than 50μg/kg except for gentisic acid (100μg/kg). Finally the method was applied to different types of feed oils, and it can be observed that higher concentrations of polyphenols were found in olive oil, whereas pomace olive oil and sunflower oil had the lowest level of these compounds.
Inflammation is a tightly coordinated response of the host immune system to bacterial and viral infections, triggered by the production of inflammatory cytokines. Sepsis is defined as a systemic ...inflammatory response followed by immunosuppression of the host and organ dysfunction. This imbalance of the immune response increases the risk of mortality of patients with sepsis, making it a major problem for critical care units worldwide. The P2X7 receptor plays a crucial role in activating the immune system by inducing the activation of peripheral blood mononuclear cells. In this study, we analyzed a cohort of abdominal origin septic patients and found that the expression of the P2X7 receptor in the plasma membrane is elevated in the different subsets of lymphocytes. We observed a direct relationship between the percentage of P2X7-expressing lymphocytes and the early inflammatory response in sepsis. Additionally, in patients whose lymphocytes presented a higher percentage of P2X7 surface expression, the total lymphocytes populations proportionally decreased. Furthermore, we found a correlation between elevated soluble P2X7 receptors in plasma and inflammasome-dependent cytokine IL-18. In summary, our work demonstrates that P2X7 expression is highly induced in lymphocytes during sepsis, and this correlates with IL-18, along with other inflammatory mediators such as IL-6, IL-8, and procalcitonin.
The diffusion of multidrug-resistant (MDR) bacteria has created the need to identify risk factors for acquiring resistant pathogens in patients living in the community.
To analyze clinical features ...of patients with community-onset pneumonia due to MDR pathogens, to evaluate performance of existing scoring tools and to develop a bedside risk score for an early identification of these patients in the Emergency Department.
This was an open, observational, prospective study of consecutive patients with pneumonia, coming from the community, from January 2011 to January 2013. The new score was validated on an external cohort of 929 patients with pneumonia admitted in internal medicine departments participating at a multicenter prospective study in Spain.
A total of 900 patients were included in the study. The final logistic regression model consisted of four variables: 1) one risk factor for HCAP, 2) bilateral pulmonary infiltration, 3) the presence of pleural effusion, and 4) the severity of respiratory impairment calculated by use of PaO2/FiO2 ratio. A new risk score, the ARUC score, was developed; compared to Aliberti, Shorr, and Shindo scores, this point score system has a good discrimination performance (AUC 0.76, 95% CI 0.71-0.82) and calibration (Hosmer-Lemeshow, χ2 = 7.64; p = 0.469). The new score outperformed HCAP definition in predicting etiology due to MDR organism. The performance of this bedside score was confirmed in the validation cohort (AUC 0.68, 95% CI 0.60-0.77).
Physicians working in ED should adopt simple risk scores, like ARUC score, to select the most appropriate antibiotic regimens. This individualized approach may help clinicians to identify those patients who need an empirical broad-spectrum antibiotic therapy.
A new method has been developed and validated for the simultaneous determination of four biogenic (putrescine, cadaverine, histamine, and tyramine) and three volatile amines (trimethylamine, ...triethylamine, and tripropylamine) in anchovy. Separation and determination of the selected compounds were carried out by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), using an electrospray ionization source (ESI) in positive ion mode. Chromatographic separation was carried out using an aqueous solution of formic acid (0.1%) and methanol as mobile phase in gradient mode. The method was validated, and mean recoveries were evaluated at three concentration levels (75, 150, and 250 μg/kg), ranging from 70 to 110% at the three levels assayed. Intra- and interday precision, expressed as relative standard deviation (RSD), were lower than 15% and 20%, respectively. Limits of quantitation (LOQs) were 25 μg/kg for all cases, except for that of TMA, which was set at 60 μg/kg. The developed procedure was applied to determine the target compounds in anchovy samples stored during 7 days at 4 °C, observing the increasing in the concentration of these compounds at longer storage time.
The resolution of fluorescence microscopy images is limited by the physical properties of light. In the last decade, numerous super-resolution microscopy (SRM) approaches have been proposed to deal ...with such hindrance. Here we present Mean-Shift Super Resolution (MSSR), a new SRM algorithm based on the Mean Shift theory, which extends spatial resolution of single fluorescence images beyond the diffraction limit of light. MSSR works on low and high fluorophore densities, is not limited by the architecture of the optical setup and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image stacks, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other SRM approaches. Along with its wide accessibility, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.
Sepsis is characterized by a systemic inflammatory response followed by immunosuppression of the host. Metabolic defects and mitochondrial failure are common in immunocompromised patients with ...sepsis. The NLRP3 inflammasome is important for establishing an inflammatory response after activation by the purinergic P2X7 receptor. Here, we study a cohort of individuals with intra-abdominal origin sepsis and show that patient monocytes have impaired NLRP3 activation by the P2X7 receptor. Furthermore, most sepsis-related deaths are among patients whose NLRP3 activation is profoundly altered. In monocytes from sepsis patients, the P2X7 receptor is associated with mitochondrial dysfunction. Furthermore, activation of the P2X7 receptor results in mitochondrial damage, which in turn inhibits NLRP3 activation by HIF-1α. We show that mortality increases in a mouse model of sepsis when the P2X7 receptor is activated in vivo. These data reveal a molecular mechanism initiated by the P2X7 receptor that contributes to NLRP3 impairment during infection.
•Comparison of the levels of phytochemicals in fresh and fresh-cut vegetables.•Fresh and fresh-cut vegetables have similar content in phytochemicals.•Multivariate analysis was used to distinguish ...between fresh and fresh-cut samples.
Bearing in mind that fresh-cut market is currently the fastest growing subsector in the food industry, a comparison of the levels of phytochemicals in fresh and fresh-cut vegetables has been carried out. Thus, several families of phytochemicals, such as phenolic acids, isoflavones, flavones, flavonols and glucosinolates were determined in fresh and fresh-cut samples including tomato, carrot, grape, eggplant and broccoli. Both type of products have potential and similar beneficial properties, regarding its content as phytochemicals, except tomato, which should be consumed as fresh. Other factors such as commercial presentation (sliced, grated, diced) and storage conditions (temperature and light) were evaluated observing that in eggplant, the content of phenolic acids is statistically different depending on the presentation. On the other hand, the content of phytochemicals was higher when fresh-cut carrots were stored at 4°C regardless of the presence or absence of light. Multivariate analysis, based on cluster analysis was used as a first approach to distinguish between fresh and fresh-cut samples, obtaining good results except for eggplant and carrot.
•An UHPLC–MS/MS method was developed to quantify several families of phytochemicals.•Analysis of phytochemicals in broccoli, tomato, grape, carrot and eggplant.•Significant differences of ...phytochemicals between analysed samples were found.
In this study a simultaneous determination of several classes of phytochemicals (isoflavones, glucosinolates, flavones, flavonols and phenolic acids) in tomato, broccoli, carrot, eggplant and grape has been carried out by ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS). Solid–liquid extraction assisted by rotary agitator was utilised, using a mixture of methanol:water (80:20, v/v) as solvent. The analytical procedure was validated in all the matrices, obtaining recoveries ranging from 60% to 120% with repeatability values (expressed as relative standard deviations, RSDs) lower than 25%. Limits of quantification (LOQs) were always equal or lower than 50μg/kg, except for some glucosinolates (125μg/kg). Finally the method was applied to different matrices such as tomato, broccoli, carrot, grape and eggplant, observing that chlorogenic acid was detected in most of the samples at higher concentrations in relation to the other compounds.
Zn, Cu, and Fe concentrations were measured in dehydrated herbs (thyme, rosemary, cloves, oregano, and basil) marketed in bulk or packaged in glass or polyethylene terephthalate (PET). Microbial ...counts of Listeria monocytogenes and other five foodborne pathogens were also checked when herbs were previously added to the growing media. The highest mean concentrations were found in basil for Zn and Cu, and in thyme and basil for Fe; the lowest ones for these minerals were in cloves (p < 0.05). Basil had significantly higher microbial counts in five of the six foodborne pathogens studied (p < 0.05). Cloves have the best hygienic quality as there is no microbial growth of L. monocytogenes, Clostridium perfringens, and Bacillus cereus; they therefore could be used as a natural preservative in food. Aromatic herbs marketed in bulk showed a significantly higher microbial count (p < 0.05). Zn, Cu, and Fe concentrations were positively correlated with microbial growth for L. monocytogenes, C. perfringens, B. cereus, and psychrophilic microorganisms (p < 0.05), so they could act as a growing factor for the foodborne pathogens.
Ca and Mg levels were determined in five spices according to marketing system (in bulk or commercialized in glass or polyethylene terephthalate (PET) containers) and correlated with microbial growth ...of commonly sporulated (Clostridium perfringens and Bacillus cereus) and non-sporulated (Listeria monocytogenes, psychrophilic and mesophilic bacteria, and yeasts and molds) food-borne pathogens present in them, when they were previously added to the microbial culture media. The basil had the highest mean Ca and Mg level and showed the highest microbial growth in the food-borne pathogenic microorganisms studied (p < 0.001). For Ca, the lowest levels were measured in cloves (p < 0.001), which had the lowest capacity for microbial contamination. Ca and Mg contents in spices correlated linear and positively (p < 0.05). Ca concentrations weakly and positively correlated (p < 0.05) with microbial counts for almost all studied microorganisms, and Mg levels for B. cereus, C. perfringens, and mesophilic bacteria (p < 0.05), possibly acting as a growing factor for some sporulated and non-sporulated foodborne pathogens. These relationships are especially significant when PET vs. glass was used as a packaging material for spices.
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