Environmental chemicals change extracellular lipidome of mature human white adipocytes

Funding This project has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement GOLIATH grant No. 825489 and Swedish Research Council for Sustainable Development (FORMAS) grant No. 2019-00375.Peer reviewedPublisher PD

A choline-releasing glycerophosphodiesterase essential for phosphatidylcholine biosynthesis and blood stage development in the malaria parasite.

The malaria parasite Plasmodium falciparum synthesizes significant amounts of phospholipids to meet the demands of replication within red blood cells. De novo phosphatidylcholine (PC) biosynthesis via the Kennedy pathway is essential, requiring choline that is primarily sourced from host serum lysophosphatidylcholine (lysoPC). LysoPC also acts as an environmental sensor to regulate parasite sexual differentiation. Despite these critical roles for host lysoPC, the enzyme(s) involved in its breakdown to free choline for PC synthesis are unknown. Here we show that a parasite glycerophosphodiesterase (PfGDPD) is indispensable for blood stage parasite proliferation. Exogenous choline rescues growth of PfGDPD-null parasites, directly linking PfGDPD function to choline incorporation. Genetic ablation of PfGDPD reduces choline uptake from lysoPC, resulting in depletion of several PC species in the parasite, whilst purified PfGDPD releases choline from glycerophosphocholine in vitro. Our results identify PfGDPD as a choline-releasing glycerophosphodiesterase that mediates a critical step in PC biosynthesis and parasite survival

Impact of Flavonols on Cardiometabolic Biomarkers: A Meta-Analysis of Randomized Controlled Human Trials to Explore the Role of Inter-Individual Variability

Several epidemiological studies have linked flavonols with decreased risk of cardiovascular disease (CVD). However, some heterogeneity in the individual physiological responses to the consumption of these compounds has been identified. This meta-analysis aimed to study the effect of flavonol supplementation on biomarkers of CVD risk such as, blood lipids, blood pressure and plasma glucose, as well as factors affecting their inter-individual variability. Data from 18 human randomized controlled trials were pooled and the effect was estimated using fixed or random effects meta-analysis model and reported as difference in means (DM). Variability in the response of blood lipids to supplementation with flavonols was assessed by stratifying various population subgroups: age, sex, country, and health status. Results showed significant reductions in total cholesterol (DM = -0.10 mmol/L; 95% CI: -0.20, -0.01), LDL cholesterol (DM = -0.14 mmol/L; Nutrients 2017, 9, 117 2 of 21 95% CI: -0.21, 0.07), and triacylglycerol (DM = -0.10 mmol/L; 95% CI: -0.18, 0.03), and a significant increase in HDL cholesterol (DM = 0.05 mmol/L; 95% CI: 0.02, 0.07). A significant reduction was also observed in fasting plasma glucose (DM = -0.18 mmol/L; 95%CI: -0.29, -0.08), and in blood pressure (SBP: DM = -4.84 mmHg; 95% CI: -5.64, -4.04; DBP: DM = -3.32 mmHg; 95% CI: -4.09, -2.55). Subgroup analysis showed a more pronounced effect of flavonol intake in participants from Asian countries and in participants with diagnosed disease or dyslipidemia, compared to healthy and normal baseline values. In conclusion, flavonol consumption improved biomarkers of CVD risk, however, country of origin and health status may influence the effect of flavonol intake on blood lipid levels

Impact of flavonols on cardiometabolic biomarkers: A meta-analysis of randomized controlled human trials to explore the role of inter-individual variability, review article

Several epidemiological studies have linked flavonols with decreased risk of cardiovascular disease (CVD). However, some heterogeneity in the individual physiological responses to the consumption of these compounds has been identified. This meta-analysis aimed to study the effect of flavonol supplementation on biomarkers of CVD risk such as, blood lipids, blood pressure and plasma glucose, as well as factors affecting their inter-individual variability. Data from 18 human randomized controlled trials were pooled and the effect was estimated using fixed or random effects meta-analysis model and reported as difference in means(DM). Variability in the response of blood lipids to supplementation with flavonols was assessed by stratifying various population subgroups: age, sex, country, and health status. Results showed significant reductions in total cholesterol (DM = 0.10 mmol/L; 95% CI: 0.20, 0.01), LDL cholesterol (DM = 0.14 mmol/L; 95% CI: 0.21, 0.07), and triacylglycerol (DM = 0.10 mmol/L; 95% CI: 0.18, 0.03), and a significant increase in HDL cholesterol (DM = 0.05 mmol/L; 95% CI: 0.02, 0.07). A significant reduction was also observed in fasting plasma glucose (DM = 0.18 mmol/L; 95% CI: 0.29, 0.08), and in blood pressure (SBP: DM = 4.84 mmHg; 95% CI: 5.64, 4.04; DBP: DM = 3.32 mmHg; 95% CI: 4.09, 2.55). Subgroup analysis showed a more pronounced effect of flavonol intake in participants from Asian countries and in participants with diagnosed disease or dyslipidemia, compared to healthy and normal baseline values. In conclusion, flavonol consumption improved biomarkers of CVD risk,however, country of origin and health status may influence the effect of flavonol intake on blood lipid levels

<b>MODERATE-INTENSITY COMBINED TRAINING INDUCES LIPIDOMIC CHANGES IN INDIVIDUALS WITH OBESITY AND TYPE 2 DIABETES</b>

Abstract: Context Alterations in the lipid metabolism are linked to metabolic disorders such as insulin resistance (IR), obesity and type 2 diabetes (T2D). Regular exercise, particularly combined training (CT), is a well-known non-pharmacological treatment that combines aerobic (AT) and resistance (RT) training benefits. However, it is unclear whether moderate-intensity exercise without dietary intervention induces changes in lipid metabolism to promote a 'healthy lipidome'. Objective The study aimed to investigate the effect of 16 weeks of CT on plasma and white adipose tissue in both sexes, middle-aged subjects with normal weight, obesity and T2D using an ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) untargeted lipidomics approach. Methods Body composition, maximum oxygen consumption (VO2 max), strength, and biochemical markers were evaluated before and after the control/training period and correlated with lipid changes. CT consisted of 8 to 10 RT exercises, followed by 35 min of AT (45 -70% VO2 max), 3 times a week for 16 weeks. Results The CT significantly reduced the levels of saturated and monounsaturated fatty acid side-chains (SFA/MUFA) in sphingolipids, glycerolipids (GL) and glycerophospholipids (GP) as well as reducing fat mass, circumferences and IR. Increased levels of polyunsaturated fatty acids in GPs, and GLs were also observed, along with increased fat-free mass, VO2 max, and strength (all p < 0.05) after training. Conclusion Our study stated that 16 weeks of moderate-intensity CT remodelled the lipid metabolism in OB, and T2D individuals, even without dietary intervention, establishing a link between exercise-modulated lipid markers and mechanisms that reduce IR and obesity-related comorbidities.</p

ZDHHC_metabolomics_chemgen_2023.zip

MetabolomicsAcyl- and probe-Coenzyme A analysis.HEK293T cells were seeded in 6-well plates, grown to 70% confluency in media containing 0.5% FBS and treated with 30 mM YnPal or 18-Bz for 2 h. Cells were dislodged into their growth media by pipetting and pelleted by centrifugation (500 x g, 5 min). The cell pellet was washed twice by resuspending in ice-cold PBS and pelleting by centrifugation.Sample extractionTo each sample, 400 µL chloroform was added and vortexed for ~1 min, followed by addition of 200 µL methanol and a repeated vortex. Samples were incubated in a water bath sonicator (4°C, 1 h), with 3 × 8 min sonication pulses, followed by centrifugation (4°C, 10 min, 17,000 x g). The supernatant was transferred to a new Eppendorf 1.5 mL tube (E1). The pellet was re-extracted with 450 µL methanol:water (2:1 v:v, containing internal standard, 13C3-Malonyl-CoA), sonicated (8 min, 4°C) and centrifuged, as above. The supernatant was added to the first extract (E1). Combined extracts were dried using a speedvac concentrator, re-suspended in 350 µL chloroform:methanol:water (1:3:3, v/v), and centrifuged, as above. The upper, aqueous phase containing the polar metabolites (including probe, probe-CoA, and acyl-CoA molecules) was dried using the speedvac concentrator and resuspended in 100 µL acetonitrile/ammonium carbonate 20 mM (7:3, v/v) for LC-MS injection.Liquid chromatography-mass spectrometry (LC-MS)Chromatography conditions:Chromatography prior to all mass spectrometry was performed using an adaptation of a method previously described61. Samples were injected into a Dionex UltiMate 3000 LC system (Thermo Fisher) with a Phenomenex Luna C18(2) 100 Å (100 x 2 mm, 3 μm) column coupled with a SecurityGuard C18 guard column (4 x 2 mm). Analytes were separated using 20 mM ammonium carbonate in water (Optima HPLC grade, Sigma Aldrich) as solvent A and acetonitrile (Optima HPLC grade, Sigma Aldrich) as solvent B at 0.3 mL/min flow rate. Elution began at 5% Solvent B, maintained for 3 min, increased to 100% B over 12 min, followed by a 3 min wash of 100% B and subsequent 3 min re-equilibration to 5% B. Other parameters were as follows: column temperature, 30°C; injection volume, 10 μL; needle wash, 50% methanol; autosampler temperature, 4°C.High resolution mass spectrometryPost-chromatography, high resolution (HR) MS was performed with positive and negative polarity switching using a Q-Exactive Orbitrap (Thermo Fisher) with a HESI-II (Heated electrospray ionization) probe. MS parameters were as follows: spray voltage, 3.5 kV and 3.2 kV (for positive and negative modes, respectively); probe temperature, 320°C; sheath and auxiliary gases, 30 and 5 arbitrary units (au), respectively; full scan range: 100 to 1300 m/z with settings of AGC target and resolution as Balanced and High (3 × 106 and 70,000), respectively. Data were recorded using Xcalibur 3.0.63 software (Thermo Fisher). Mass calibration was performed for both ESI polarities before analysis using the standard Thermo Fisher Calmix solution. Qualitative analysis was performed using Xcalibur FreeStyle 1.8 SP1 and Tracefinder 5.1 software (Thermo Fisher) according to the manufacturer’s workflows. Masses, retention times, and fragmentation of all relevant sample-derived molecules were compared to authentic chemical standards.MS/MS MS parameters were optimized by direct infusion of 16 μM acyl-CoAs dissolved in 10 mM MeOH/ammonium acetate at 5 μL/min into an TSQ Quantiva triple quadrupole MS (Thermo Fisher). The heated electrospray was set in positive mode with the following parameters: capillary voltage, 3472 V; sheath gas, 60 au; aux gas, 10 au; sweep gas, 1 au; ion transfer tube temp, 325°C; vaporizer temp, 275°C. A selected reaction monitoring (SRM) function was applied for the simultaneous detection of acyl-CoA and probe-CoA molecules with RF lens and collision energies as shown in the Supplementary Table 7. Data were recorded using the Xcalibur 4.0.27.10 software and analysed using QuanBrowser 4.5.445.18 (Thermo Fisher).</p

Impact&nbsp;of&nbsp;Flavonols&nbsp;on&nbsp;Cardiometabolic&nbsp;Biomarkers:&nbsp; A&nbsp;Meta‐Analysis&nbsp;of&nbsp;Randomized&nbsp;Controlled&nbsp;Human&nbsp; Trials&nbsp;to&nbsp;Explore&nbsp;the&nbsp;Role&nbsp;of&nbsp;Inter‐Individual&nbsp; Variability

Several&nbsp; epidemiological&nbsp; studies&nbsp; have&nbsp; linked&nbsp; flavonols&nbsp; with&nbsp; decreased&nbsp; risk&nbsp; of&nbsp; cardiovascular&nbsp; disease&nbsp; (CVD).&nbsp; However,&nbsp; some&nbsp; heterogeneity&nbsp; in&nbsp; the&nbsp; individual&nbsp; physiological&nbsp; responses&nbsp;to&nbsp;the&nbsp;consumption&nbsp;of&nbsp;these&nbsp;compounds&nbsp;has&nbsp;been&nbsp;identified.&nbsp;This&nbsp;meta-analysis&nbsp;aimed&nbsp;to&nbsp; study&nbsp;the&nbsp;effect&nbsp;of&nbsp;flavonol&nbsp;supplementation&nbsp;on&nbsp;biomarkers&nbsp;of&nbsp;CVD&nbsp;risk&nbsp;such&nbsp;as,&nbsp;blood&nbsp;lipids,&nbsp;blood&nbsp; pressure&nbsp;and&nbsp;plasma&nbsp;glucose,&nbsp;as&nbsp;well&nbsp;as&nbsp;factors&nbsp;affecting&nbsp;their&nbsp;inter-individual&nbsp;variability.&nbsp;Data&nbsp;from&nbsp; 18&nbsp;human&nbsp;randomized&nbsp;controlled&nbsp;trials&nbsp;were&nbsp;pooled&nbsp;and&nbsp;the&nbsp;effect&nbsp;was&nbsp;estimated&nbsp;using&nbsp;fixed&nbsp;or&nbsp; random&nbsp;effects&nbsp;meta-analysis&nbsp;model&nbsp;and&nbsp;reported&nbsp;as&nbsp;difference&nbsp;in&nbsp;means&nbsp;(DM).&nbsp;Variability&nbsp;in&nbsp;the&nbsp; response&nbsp;of&nbsp;blood&nbsp;lipids&nbsp;to&nbsp;supplementation&nbsp;with&nbsp;flavonols&nbsp;was&nbsp;assessed&nbsp;by&nbsp;stratifying&nbsp;various&nbsp; population&nbsp;subgroups:&nbsp;age,&nbsp;sex,&nbsp;country,&nbsp;and&nbsp;health&nbsp;status.&nbsp;Results&nbsp;showed&nbsp;significant&nbsp;reductions&nbsp; in&nbsp;total&nbsp;cholesterol&nbsp;(DM&nbsp;=&nbsp;-0.10&nbsp;mmol/L;&nbsp;95% CI:&nbsp;-0.20,&nbsp;-0.01),&nbsp;LDL&nbsp;cholesterol&nbsp;(DM&nbsp;=&nbsp;-0.14&nbsp;mmol/L;&nbsp; Nutrients&nbsp;2017,&nbsp;9,&nbsp;117&nbsp; 2&nbsp;of&nbsp;21&nbsp; 95% CI:&nbsp;-0.21,&nbsp;0.07),&nbsp;and&nbsp;triacylglycerol&nbsp;(DM&nbsp;=&nbsp;-0.10&nbsp;mmol/L;&nbsp;95% CI:&nbsp;-0.18,&nbsp;0.03),&nbsp;and&nbsp;a&nbsp;significant&nbsp; increase&nbsp;in&nbsp;HDL&nbsp;cholesterol&nbsp;(DM&nbsp;=&nbsp;0.05&nbsp;mmol/L;&nbsp;95% CI:&nbsp;0.02,&nbsp;0.07).&nbsp;A&nbsp;significant&nbsp;reduction&nbsp;was&nbsp;also&nbsp; observed&nbsp;in&nbsp;fasting&nbsp;plasma&nbsp;glucose&nbsp;(DM&nbsp;=&nbsp;-0.18&nbsp;mmol/L;&nbsp;95%CI:&nbsp;-0.29,&nbsp;-0.08),&nbsp;and&nbsp;in&nbsp;blood&nbsp;pressure&nbsp; (SBP:&nbsp;DM = -4.84&nbsp;mmHg;&nbsp;95% CI:&nbsp;-5.64,&nbsp;-4.04;&nbsp;DBP:&nbsp;DM = -3.32&nbsp;mmHg;&nbsp;95%&nbsp;CI: -4.09, -2.55).&nbsp; Subgroup&nbsp;analysis&nbsp;showed&nbsp;a&nbsp;more&nbsp;pronounced&nbsp;effect&nbsp;of&nbsp;flavonol&nbsp;intake&nbsp;in&nbsp;participants&nbsp;from&nbsp;Asian&nbsp; countries&nbsp;and&nbsp;in&nbsp;participants&nbsp;with&nbsp;diagnosed&nbsp;disease&nbsp;or&nbsp;dyslipidemia,&nbsp;compared&nbsp;to&nbsp;healthy&nbsp;and&nbsp; normal&nbsp;baseline&nbsp;values.&nbsp;In&nbsp;conclusion,&nbsp;flavonol&nbsp;consumption&nbsp;improved&nbsp;biomarkers&nbsp;of&nbsp;CVD&nbsp;risk,&nbsp; however,&nbsp;country&nbsp;of&nbsp;origin&nbsp;and&nbsp;health&nbsp;status&nbsp;may&nbsp;influence&nbsp;the&nbsp;effect&nbsp;of&nbsp;flavonol&nbsp;intake&nbsp;on&nbsp;blood&nbsp; lipid&nbsp;levels

Lipidomic Approaches to Study HDL Metabolism in Patients with Central Obesity Diagnosed with Metabolic Syndrome.

Funder: Foundation for Liver ResearchFunder: Royal Society and a CAMS-UK fellowshipThe metabolic syndrome (MetS) is a cluster of cardiovascular risk factors characterised by central obesity, atherogenic dyslipidaemia, and changes in the circulating lipidome; the underlying mechanisms that lead to this lipid remodelling have only been partially elucidated. This study used an integrated "omics" approach (untargeted whole serum lipidomics, targeted proteomics, and lipoprotein lipidomics) to study lipoprotein remodelling and HDL composition in subjects with central obesity diagnosed with MetS (vs. controls). Compared with healthy subjects, MetS patients showed higher free fatty acids, diglycerides, phosphatidylcholines, and triglycerides, particularly those enriched in products of de novo lipogenesis. On the other hand, the "lysophosphatidylcholines to phosphatidylcholines" and "cholesteryl ester to free cholesterol" ratios were reduced, pointing to a lower activity of lecithin cholesterol acyltransferase (LCAT) in MetS; LCAT activity (directly measured and predicted by lipidomic ratios) was positively correlated with high-density lipoprotein cholesterol (HDL-C) and negatively correlated with body mass index (BMI) and insulin resistance. Moreover, many phosphatidylcholines and sphingomyelins were significantly lower in the HDL of MetS patients and strongly correlated with BMI and clinical metabolic parameters. These results suggest that MetS is associated with an impairment of phospholipid metabolism in HDL, partially led by LCAT, and associated with obesity and underlying insulin resistance. This study proposes a candidate strategy to use integrated "omics" approaches to gain mechanistic insights into lipoprotein remodelling, thus deepening the knowledge regarding the molecular basis of the association between MetS and atherosclerosis

Impact of Flavonols on Cardiometabolic Biomarkers: A Meta-Analysis of Randomized Controlled Human Trials to Explore the Role of Inter-Individual Variability

Several epidemiological studies have linked flavonols with decreased risk of cardiovascular disease (CVD). However, some heterogeneity in the individual physiological responses to the consumption of these compounds has been identified. This meta-analysis aimed to study the effect of flavonol supplementation on biomarkers of CVD risk such as, blood lipids, blood pressure and plasma glucose, as well as factors affecting their inter-individual variability. Data from 18 human randomized controlled trials were pooled and the effect was estimated using fixed or random effects meta-analysis model and reported as difference in means (DM). Variability in the response of blood lipids to supplementation with flavonols was assessed by stratifying various population subgroups: age, sex, country, and health status. Results showed significant reductions in total cholesterol (DM = -0.10 mmol/L; 95% CI: -0.20, -0.01), LDL cholesterol (DM = -0.14 mmol/L; Nutrients 2017, 9, 117 2 of 21 95% CI: -0.21, 0.07), and triacylglycerol (DM = -0.10 mmol/L; 95% CI: -0.18, 0.03), and a significant increase in HDL cholesterol (DM = 0.05 mmol/L; 95% CI: 0.02, 0.07). A significant reduction was also observed in fasting plasma glucose (DM = -0.18 mmol/L; 95%CI: -0.29, -0.08), and in blood pressure (SBP: DM = -4.84 mmHg; 95% CI: -5.64, -4.04; DBP: DM = -3.32 mmHg; 95% CI: -4.09, -2.55). Subgroup analysis showed a more pronounced effect of flavonol intake in participants from Asian countries and in participants with diagnosed disease or dyslipidemia, compared to healthy and normal baseline values. In conclusion, flavonol consumption improved biomarkers of CVD risk, however, country of origin and health status may influence the effect of flavonol intake on blood lipid levels

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field