APOE genotype modifies the plasma oxylipin response to omega-3 polyunsaturated fatty acid supplementation in healthy individuals

The omega-3 polyunsaturated fatty acids (n-3 PUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), mediate inflammation in large part by affecting pro-inflammatory and anti-inflammatory/pro-resolving oxylipin concentrations. Common gene variants are thought to underlie the large inter-individual variation in oxylipin levels in response to n-3 PUFA supplementation, which in turn is likely to contribute to the overall heterogeneity in response to n-3 PUFA intervention. Given its known role in inflammation and as a modulator of the physiological response to EPA and DHA, here we explore, for the first time, the differential response of plasma hydroxy-, epoxy- and dihydroxy-arachidonic acid, EPA and DHA oxylipins according to apolipoprotein E (APOE) genotype using samples from a dose-response parallel design RCT. Healthy participants were given doses of EPA+DHA equivalent to intakes of 1, 2, and 4 portions of oily fish per week for 12 months. There was no difference in the plasma levels of EPA, DHA or ARA between the wildtype APOE3/E3 and APOE4 carrier groups after 3 or 12 months of n-3 PUFA supplementation. At 12 months, hydroxy EPAs (HEPEs) and hydroxy-DHAs (HDHAs) were higher in APOE4 carriers, with the difference most evident at the highest EPA+DHA intake. A significant APOE *n-3 PUFA dose effect was observed for the CYP-ω hydroxylase products 19-HEPE (p = 0.027) and 20-HEPE (p = 0.011). 8-HEPE, which, along with several other plasma oxylipins, is an activator of peroxisome proliferator activated receptors (PPARs), showed the highest fold change in APOE4 carriers (14-fold) compared to APOE3/E3 (4-fold) (p = 0.014). Low basal plasma EPA levels (EPA 1.22% of total fatty acids). In conclusion, APOE genotype modulated the plasma oxylipin response to increased EPA+DHA intake, with APOE4 carriers presenting with the greatest increases following high dose n-3 PUFA supplementation for 12 months

Development of an Optimized LC-MS Method for the Detection of Specialized Pro-Resolving Mediators in Biological Samples

The cardioprotective and anti-inflammatory effects of long chain omega-3 polyunsaturated fatty acids (n3 PUFA) are believed to be partly mediated by their oxygenated metabolites (oxylipins). In the last two decades interest in a novel group of autacoids termed specialized pro-resolving mediators (SPMs) increased. These are actively involved in the resolution of inflammation. SPMs are multiple hydroxylated fatty acids including resolvins, maresins, and protectins derived from the n3 PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as well as lipoxins derived from arachidonic acid (ARA). In the present paper, we developed an LC-MS/MS method for a comprehensive set of 18 SPMs derived from ARA, EPA, and DHA and integrated it into our targeted metabolomics platform. Quantification was based on external calibration utilizing five deuterated internal standards in combination with a second internal standard for quality assessment of sample preparation in each sample. The tandem mass spectrometric parameters were carefully optimized for sensitive and specific detection. The influence of source parameters of the used AB Sciex 6500 QTRAP instrument as well as electronic parameters and the selection of transitions are discussed. The method was validated/characterized based on the criteria listed in the European Medicines Agency (EMA) guideline on bioanalytical method validation and method performance is demonstrated regarding recovery of internal standards (between 78 ± 4% and 87 ± 3% from 500 μL of human serum) as well as extraction efficacy of SPMs in spiked plasma (intra-day accuracy within ±20 and ±15% at 0.1 and 0.3 nM in plasma, respectively). Based on the lower limit of quantification of 0.02–0.2 nM, corresponding to 0.18–2.7 pg on column, SPMs were generally not detectable/quantifiable in plasma and serum supporting that circulating levels of SPMs are very low, i.e., <0.1 nM in healthy subjects. Following septic shock or peritonitis, SPMs could be quantified in the samples of several patients. However, in these studies with a small number of patients no clear correlation with severity of inflammation could be observed

Comparison of the effects of long-chain omega-3 fatty acid supplementation on plasma levels of free and esterified oxylipins

IntroductionIt is believed that many of the beneficial effects of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) are mediated by their oxidized metabolites, the oxylipins. The formation and biological role of many cytochrome P450 and lipoxygenase derived hydroxy, epoxy and dihydroxy FA, particularly of oxylipins esterified in polar lipids and triglycerides remain unclear. In this study, we compared the impact of twelve weeks of LC n-3 PUFA supplementation on the patterns of free and total (sum of esterified and free) hydroxy, epoxy and dihydroxy FAs.Subjects and methodsSubjects (5 male; 5 female) between 46 and 70 years were supplemented with 1.1g/d of eicosapentaenoic acid (EPA) and 0.74g/d docosahexaenoic acid (DHA) as ethyl esters. Blood samples were drawn before and after twelve weeks of treatment. Oxylipins in plasma were analyzed by LC-MS directly for free oxylipins and after saponification. Relative FA composition in erythrocyte membranes was analyzed by GC.ResultsLC n-3 PUFA treatment led to a significant increase in EPA (200%) and DHA (23%) in erythrocyte membranes. Of the oxylipins measured in plasma, total and free EPA-derived metabolites were highly increased (70-150%), while total AA-derived metabolites were decreased on average by 30%. There was no effect on DHA-metabolites. Concentrations of total hydroxy and epoxy FAs in plasma were considerably higher compared to free hydroxy and epoxy FAs (up to 350 times), while levels of most free dihydroxy FAs were in a similar range to total dihydroxy FAs. However, the individual ratios between total and free plasma oxylipins remained unchanged after LC n-3 PUFA treatment.Discussion and conclusionsLC n-3 PUFA supplementation causes a shift in the levels of circulating oxylipins, having the strongest impact on EPA-derived epoxy, dihydroxy and hydroxy FA. The unchanged ratio of free and esterified oxylipins in plasma indicates that both concentrations are valuable biomarkers for assessing the individual status of these lipid mediators

Extraction of lipids and oxylipins from plasma for quantification of total oxylipins – Challenges and strategies

Several eicosanoids and other oxylipins are potent lipid mediators which are involved in theregulation of physiological functions such as inflammation. It is believed that they act predominantlyin their free, i.e. non-esterified, form. However, a major portion of oxylipins is found as esters, e.g. inpolar lipids.Only little information is available on the biological activity of these esterified oxylipins. An importantstep towards a better understanding of their biological role is a comprehensive comparison of thechanges in the pattern of free vs. esterified mediators induced by pharmacological intervention ordietary supplementation as well as during onset and progression of diseases.While several LC-MS based methods for the detection of free oxylipins have been developed,esterified oxylipins are commonly quantified as a sum of free and esterified oxylipins following basehydrolysis. However, different approaches have been described for preparation of samples beforehydrolysis and for the solid phase extraction. Here, we present a three-step strategy for thequantification of total oxylipins including extraction of total lipids, saponification to liberate esterifiedoxylipins and solid phase extraction of free oxylipins. We thoroughly investigated different liquidliquidextraction procedures and protein precipitation in terms of extraction efficiency for variouslipid classes from plasma. Moreover, to optimize sample throughout we compared extraction of freeoxylipins via solid phase extraction on cartridges with 96-well plates.In conclusion, our results emphasis the challenges related to the extraction procedures and providedifferent strategies for reliable oxylipin quantification with a focus on efficient and reproducibleextraction

Intra-individual variance of the human plasma oxylipin pattern: Low inter-day variability in fasting blood samples: Versus high variability during the day

Introduction: Several eicosanoids and other oxylipins are important lipid mediators. Reliable quantification in plasma is important to assess the state of disease, action of drugs and the biology of oxylipins. In order to monitor biological changes, low background variability of oxylipin concentrations in biological samples is essential for proper interpretation of oxylipin biology. However, only little is known about the variation in the oxylipin profile in healthy human subjects. Experimental: Inter-day variation in circulating oxylipins after overnight fasting was investigated in healthy young men on either a standardized or non-standardized diet during a (24 to) 48 h time interval. Intra-day variance was investigated during an 8 h time interval (covering breakfast and lunch meals) in men on a standardized diet with blood sampling at 0, 2, 4, 6 and 8 hours. Free oxylipins in plasma were analyzed using a targeted metabolomics platform for the quantification of 160 oxylipins from different precursors. Analytical variation was evaluated based on quality control plasma samples. Results: Free oxylipins in quality control plasma samples showed low variations (<20% for most analytes). Inter-day variations in fasting blood were in the same range, while significant differences were observed within the day (intra-day variance). Conclusion: Based on the low intra-individual inter-day variance in concentrations of free oxylipins, our results demonstrate the suitability of fasting plasma for the investigation of oxylipin biology. In non-fasting plasma samples, the variations were high during the day. Thus, non-fasting plasma samples appear to be unsuitable to evaluate biologically relevant changes, for instance, those caused by disease or drugs. However, it remains to be determined if the same standardized meal results in reproducible modulations of the oxylipin profile allowing evaluation of the oxylipin pattern during the postprandial state

Fachärztliche Unterversorgung bei Heimbewohnern – Prävalenzstudie und Hochrechnung

Background!#!Nursing home residents tend to have lower medical specialist utilization than other groups of older people; however, as yet there is little evidence whether nursing home residents obtain adequate medical specialist care. This study investigated whether nursing home residents receive adequate oral health care, ophthalmological care, otorhinolaryngologist care and neurological care. The unmet needs of the nursing home population in Germany was extrapolated.!##!Material and methods!#!Audiometry, eye examinations and oral visual inspection were performed in 409 residents from 44 nursing homes. Medical care in the previous 12 months as well as existing diagnoses were retrieved from the nursing documentation. Teams of physicians evaluated for each resident based on all collected data if the resident obtained specialist care that was adequate to the needs.!##!Results!#!Between 15% and 45% of the residents with need for medical specialist care did not receive adequate specialist care. Of all residents 27% had unmet need of specialist care in at least one of the investigated medical specialties. It is projected that up to 205,000 nursing home residents in Germany do not receive adequate medical specialist care.!##!Conclusion!#!Given a considerable proportion of nursing home residents with unmet need of specialist care, interventions should be developed that help reduce the level of unmet needs

MS-based targeted metabolomics of eicosanoids and other oxylipins: Analytical variability and interlaboratory comparison of esterified oxylipin profile

IntroductionOxylipins are potent lipid mediators involved in numerous physiological and pathological processesand their quantitative profiling has gained a lot of attention [1]. To maximize the utility of theoxylipin profiling in clinical research it is now crucial (i) to assess its analytical variability; (ii) todetermine its comparability between laboratories and (iii) to identify putative critical oxylipins. Thesethree main challenges are addressed within the EU JPI HDHL*-Oxygenate project.Technological and methodological innovationTo address the challenges stated above, a SOP was established by a reference laboratory for the MSbasedtargeted metabolomics of total oxylipins (free + esterified, ~160 oxylipins) in human plasma[2]. The intra- and inter-day variabilities of each oxylipin were assessed. Then, the SOP wastransferred to 4 independent laboratories together with mixtures of internal standards, calibrantsand 7 different pools of plasma to determine the comparability of oxylipin profiles between labs.Results and impactThe cumulated intra-/inter-day variabilities revealed that 68 % of oxylipins (>LLOQ) have a CV<20%.The interlab-variability was low and dependent on the type of plasma analyzed. Overall, our resultsshow that the MS-based profiling of total oxylipins in human plasma is a robust tool for clinicalresearch. Moreover, the comparability of oxylipin profiles will allow generating large-scale databasesallowing a better understanding of the relationships between oxylipins and human health.References[1] Gladine C. et al. 2019. Free Radical Biology and Medicine 144 (2019) 72–89[2] Ostermann et al. 2019. Prostag Oth Lipid M. DOI: 10.1016/j.prostaglandins.2019.106384*Joint Programming Initiative “A healthy diet for a healthy life

Bedarfe der Langzeitpflege in der COVID-19-Pandemie

The SARS-CoV‑2 virus and the associated disease COVID-19 pose major challenges to healthcare systems worldwide. Especially the vulnerable group of people in need of long-term care is at risk of suffering a severe course of the disease or of dying from the infection.In a nationwide cross-sectional study the situation and needs of inpatient and outpatient long-term care facilities during the SARS-CoV‑2 pandemic were assessed and analyzed using an online survey.Participants from 531 institutions postulated the need for uniform recommendations for action on SARS-CoV‑2, adequate and affordable protective and hygiene materials, serial tests in the institutions, well-founded advice on the implementation of interventions, a specific pandemic plan and supporting public relations work by the media. This calls for higher nursing remuneration, better staffing levels and greater appreciation of the nursing profession.In order to protect the vulnerable group of people in need of nursing care from a SARS-CoV‑2 infection, long-term care must be given a stronger focus in health policy measures during the pandemic

Zur Situation der Langzeitpflege in Deutschland während der Corona-Pandemie. Ergebnisse einer Online-Befragung in Einrichtungen der (teil)stationären und ambulanten Langzeitpflege

In Deutschland – aber auch weltweit – hat die COVID-19-Pandemie schwerwiegende Auswirkungen auf die Gesellschaft und insbesondere das Gesundheitssystem. Zum 2. Juni 2020 weist das Robert Koch-Institut (RKI) für Deutschland insgesamt 182.028 laborbestätigte COVID-19-Fälle aus, darunter 8.522 Todesfälle in Zusammenhang mit COVID-19. 86 % dieser Todesfälle und 16 % aller infizierten Personen waren dabei 70 Jahre oder älter. Vor allem ältere und pflegebedürftige Menschen sind bei einer Infektion mit dem SARS-CoV2-Virus von schweren Krankheitsverläufen und einer hohen Mortalität betroffen. Pflegekräfte wiederum haben aufgrund des direkten Kontaktes mit Pflegebedürftigen ein hohes Risiko an COVID-19 zu erkranken. Aufgrund der Vielzahl potentiell Betroffener (bundesweit 3,4 Mio Pflegebedürftige und 1,2 Mio. Beschäftigte in Einrichtungen der stationären und ambulanten Langzeitpflege) sollte in der vorliegenden Pandemie-Situation daher ein besonderes Augenmerk auf den Bereichen der ambulanten und stationären Langzeitpflege liegen – auch mit Blick auf eine mögliche zweie Pandemie-Welle. Derzeit fehlen jedoch valide Daten zur Situation von Einrichtungen der ambulanten und stationären Langzeitpflege, die es Leistungserbringern aber auch politischen Akteuren ermöglichen, auf die Herausforderungen und Unterstützungsbedarfe aufgrund der Pandemie organisatorisch und administrativ zu reagieren. Die Universität Bremen hat daher vom 28. April bis zum 12. Mai 2020 eine deutschlandweite Online-Befragung in (teil-)stationären und ambulanten Pflegeeinrichtungen durchgeführt. Knapp 18.000 Einrichtungen wurden mit Fragen zu Strukturmerkmalen, zum Vorkommen des SARS-CoV-2-Virus in den Einrichtungen, Auswirkungen der Pandemie z. B. in Bezug auf personelle und sachliche Ausstattung aber auch zu veränderten Arbeitsprozessen und Kommunikationsstrukturen per Email angeschrieben und zusätzlich über Verbände auf die Umfrage hingewiesen. Von 701 Pflegediensten, 96 teilstationären und 824 stationären Einrichtungen liegen nunmehr Daten vor. In Bezug auf zentrale Strukturmerkmale entspricht die Verteilung der beteiligten Einrichtungen dabei im Wesentlichen der bundesweiten Verteilung