Development and Validation of Methodologies for the Identification of Specialized Pro-Resolving Lipid Mediators and Classic Eicosanoids in Biological Matrices.

Lipid mediators, which include specialized pro-resolving mediators and classic eicosanoids, are pivotal in both initiating and resolving inflammation. The regulation of these molecules determines whether inflammation resolves naturally or persists. However, our understanding of how these mediators are regulated over time in various inflammatory contexts is limited. This gap hinders our grasp of the mechanisms underlying the disease onset and progression. Due to their localized action and low endogenous levels in many tissues, developing robust and highly sensitive methodologies is imperative for assessing their endogenous regulation in diverse inflammatory settings. These methodologies will help us gain insight into their physiological roles. Here, we establish methodologies for extracting, identifying, and quantifying these mediators. Using our methods, we identified a total of 37 lipid mediators. Additionally, by employing a reverse-phase HPLC method, we successfully separated both double-bond and chiral isomers of select lipid mediators, including Lipoxin (LX) A4, 15-epi-LXA4, Protectin (PD) D1, PDX, and 17R-PD1. Validation of the method was performed in both solvent and surrogate matrix for linearity of the standard curves, lower limits of quantitation (LLOQ), accuracy, and precision. Results from these studies demonstrated that linearity was good with r2 values > 0.98, and LLOQ for the mediators ranged from 0.01 to 0.9 pg in phase and from 0.1 to 8.5 pg in surrogate matrix. The relative standard deviation (RSD) for inter- and intraday precision in solvent ranged from 5% to 12% at low, intermediate, and high concentrations, whereas the RSD for the inter- and intraday variability in the accuracy ranged from 95% to 87% at low to high concentrations. The recovery in biological matrices (plasma and serum) for the internal standards used ranged from 60% to 118%. We observed a marked ion suppression for molecules evaluated in negative ionization mode, while there was an ion enhancement effect by the matrix for molecules evaluated in positive ionization mode. Comparison of the integration algorithms, namely, AutoPeak and MQ4, and approaches for calculating signal-to-noise ratios (i.e., US Pharmacopeia, relative noise, peak to peak, and standard deviation) demonstrated that different integration algorithms tested had little influence on signal-to-noise ratio calculations. In contrast, the method used to calculate the signal-to-noise ratio had a more significant effect on the results, with the relative noise approach proving to be the most robust. The methods described herein provide a platform to study the SPM and classic eicosanoids in biological tissues that will help further our understanding of disease mechanisms.

Effects of COVID-19-related psychological distress and anxiety on quality of sleep and life in healthcare workers in Iran and three European countries.

Introduction: The COVID-19 pandemic has considerably affected human beings most of whom are healthcare workers (HCWs) combating the disease in the front line. Methods: This cross-sectional study aims to explore the effects of stress and anxiety caused by COVID-19 on the quality of sleep and life in HCWs, including physicians, nurses, and other healthcare staff. In this global study, we asked 1,210 HCWs (620 and 590 volunteers from Iran and European countries, including Germany, the Netherlands, and Italy, respectively), who age 21-70, to participate in the test. Several measures of COVID-related stress, anxiety, sleep, and life quality, including the 12-item General Health Questionnaire (GHQ-12), Fear of COVID-19 scale (FCV-19S), Beck Anxiety Inventory (BAI), the Pittsburgh Sleep Quality Index (PSQI), and World Health Organization Quality of Life-BREF (WHOQOL-BREF) are recorded. Results: Volunteers reported high rates of stress and anxiety and poor sleep quality as well as lower quality of life. The correlation analysis between the measures is reported. According to the results, regardless of the location, HCWs, predominantly female nurses, developed anxiety and stress symptoms which consequently resulted in lower sleep and life quality. Both for Iranian and the European HCWs, significant differences existed between nurses and the other two groups, with the p-values equal to 0.0357 and 0.0429 for GHQ-12, 0.0368, and 0.714 for BAI measure. Even though nurses reported the most stress, anxiety, fear of COVID-19, lower quality of life and sleep in both countries, and also an increase in other measures as well, there existed no statistically significant difference in FCV-19S, PSQI, and WHOQOL-BREF. Discussion: This study helps to expand our knowledge the effects of pandemics on HCWs and also for healthcare management to predict HCW's mental health conditions in similar situations.