Determining the mechanisms of dietary turnip rapeseed oil on cholesterol metabolism in men with metabolic syndrome

We have earlier reported the reduction of total cholesterol low-density lipoprotein (LDL) cholesterol and oxidized LDL caused by short-term modification of diet with cold-pressed turnip rapeseed oil (CPTRO) instead of butter. The aim of this supplementary study was to determine whether the beneficial effects resulted from altered cholesterol metabolism during the intervention. Thirty-seven men with metabolic syndrome (MetS) completed an open, randomized and balanced crossover study. Subjects' usual diet was supplemented with either 37.5 g of butter or 35 mL of CPTRO for 6-8 weeks. Otherwise normal dietary habits and physical activity were maintained without major variations. Serum non-cholesterol sterols were assayed with gas-liquid chromatography and used as surrogate markers of whole-body cholesterol synthesis and absorption efficiency. Serum proprotein convertase subtilisin/kexin type 9 (PCSK9) concentration was analyzed with Quantikine ELISA Immunoassay. Serum cholesterol synthesis markers and serum cholestanol (absorption marker), all as ratios to cholesterol, did not differ between the periods. Serum campesterol and sitosterol ratios to cholesterol were significantly increased after the administration of CPTRO resulting from the increased intake of 217 mg/day of plant sterols in CPTRO. Serum PCSK9 concentration did not differ between CPTRO and butter periods. The reduction in serum cholesterol by 7.2% after consumption of rapeseed oil could not be explained by changes in cholesterol absorption, synthesis or PCSK9 metabolism in MetS.Peer reviewe

Dietary Fatty Acids and Inflammation : Observational and Interventional Studies

Dietary fat quality influences the risk of type 2 diabetes and cardiovascular disease. A low-grade inflammation is suggested to contribute to the disease development, often accompanied by obesity. Whereas n-3 polyunsaturated fatty acids (PUFA) have been considered anti-inflammatory, n-6 PUFA have been proposed to act pro-inflammatory. Saturated fatty acids (SFA) act pro-inflammatory in vitro. This thesis aimed to investigate effects of different fatty acids on low-grade inflammation in observational and interventional studies. In Paper I and II, fatty acid composition in serum cholesterol esters was used as objective marker of dietary fat quality and related to serum C-reactive protein (CRP) and other circulating inflammatory markers in two population-based cohorts, conducted in middle-aged men and elderly men and women, respectively. In Paper III and IV, the impact of diets differing in fat quality on inflammation and oxidative stress was investigated in randomised controlled studies, in subjects with metabolic syndrome and abdominal obesity. In Paper I and II, a low proportion of linoleic acid (18:2 n-6) in serum was associated with higher CRP concentrations, indicating that a low intake of vegetable fats may be related to low-grade inflammation. High CRP concentrations were also associated with high proportions of palmitoleic (16:1) and oleic (18:1) acids and high stearoyl coenzymeA desaturase index, possibly reflecting altered fat metabolism and/or high SFA intake in this population. When comparing two high-fat diets rich in either saturated or monounsaturated fat, and two low-fat diets with or without long-chain n-3 PUFA supplementation during 12 weeks (Paper III), no differences in inflammation or oxidative stress markers were observed. Moreover, a 10-week intervention (Paper IV) with high linoleic acid intake showed no adverse effects on inflammation or oxidative stress. Instead, interleukin-1 receptor antagonist and tumor necrosis factor receptor-2 decreased after linoleic acid intake compared with a diet high in SFA. The results in this thesis indicate that dietary n-6 PUFA found in vegetable fats is associated with lower inflammation marker levels, and to some extent reduces systemic inflammation when compared with SFA. Supplementation of n-3 PUFA did not exert any systemic anti-inflammatory effects, maybe due to a relatively low dose

Dietary Fatty Acids and Inflammation : Observational and Interventional Studies

Dietary fat quality influences the risk of type 2 diabetes and cardiovascular disease. A low-grade inflammation is suggested to contribute to the disease development, often accompanied by obesity. Whereas n-3 polyunsaturated fatty acids (PUFA) have been considered anti-inflammatory, n-6 PUFA have been proposed to act pro-inflammatory. Saturated fatty acids (SFA) act pro-inflammatory in vitro. This thesis aimed to investigate effects of different fatty acids on low-grade inflammation in observational and interventional studies. In Paper I and II, fatty acid composition in serum cholesterol esters was used as objective marker of dietary fat quality and related to serum C-reactive protein (CRP) and other circulating inflammatory markers in two population-based cohorts, conducted in middle-aged men and elderly men and women, respectively. In Paper III and IV, the impact of diets differing in fat quality on inflammation and oxidative stress was investigated in randomised controlled studies, in subjects with metabolic syndrome and abdominal obesity. In Paper I and II, a low proportion of linoleic acid (18:2 n-6) in serum was associated with higher CRP concentrations, indicating that a low intake of vegetable fats may be related to low-grade inflammation. High CRP concentrations were also associated with high proportions of palmitoleic (16:1) and oleic (18:1) acids and high stearoyl coenzymeA desaturase index, possibly reflecting altered fat metabolism and/or high SFA intake in this population. When comparing two high-fat diets rich in either saturated or monounsaturated fat, and two low-fat diets with or without long-chain n-3 PUFA supplementation during 12 weeks (Paper III), no differences in inflammation or oxidative stress markers were observed. Moreover, a 10-week intervention (Paper IV) with high linoleic acid intake showed no adverse effects on inflammation or oxidative stress. Instead, interleukin-1 receptor antagonist and tumor necrosis factor receptor-2 decreased after linoleic acid intake compared with a diet high in SFA. The results in this thesis indicate that dietary n-6 PUFA found in vegetable fats is associated with lower inflammation marker levels, and to some extent reduces systemic inflammation when compared with SFA. Supplementation of n-3 PUFA did not exert any systemic anti-inflammatory effects, maybe due to a relatively low dose

Concentrations of UV-filters (benzophenones) and arsenic in urine samples from participants in the dietary survey Riksmaten adolescents 2016-17

Inom ramen för Horizon 2020-projektet HBM4EU analyserade 300 urinprover från deltagare i matvane-undersökningen Riksmaten ungdom 2016-17 avseende UV-filter (bensofenoner) samt arsenik och arsenikföreningar. I denna rapport redovisas resultaten från dessa analyser och dessutom undersöks skillnader i exponering mellan pojkar och flickor och mellan åldersgrupper (årskurs 5, årskurs 8 och årskurs 2 på gymnasiet).Bensofenon-1 (BP-1) och bensofenon-3 (BP-3) kunde detekteras i nästan alla prover (292 respektive 283 av 300), medan bensofenon-2 och bensofenon-7 låg under detektionsgränsen i mer än 90% av proverna. Medianhalten för BP-1 var 0,67 ng/ml (0,49 μg/g kreatinin) och för BP-3 1,2 ng/ml (0,92 μg/g kreatinin). Halterna låg på ungefär samma nivå som i andra europeiska studier av barn och ungdomar. Flickor hade signifikant högre halter av BP-1 och BP-3 i urin än pojkar. Skillnaden kan bero på att flickor använder mer produkter innehållande bensofenoner (kosmetika och hudvårdsprodukter) än pojkar. Halterna av bensofenoner skiljde sig inte åt mellan åldersgrupperna.Förutom total arsenik (total As) analyserades oorganisk arsenik (arsenit, AsIII och arsenat, AsV), metaboliterna monometylarsonsyra (MMA) och dimetylarsinsyra (DMA) samt den organiska formen arsenobetain i urin. Samtliga föreningar kunde kvantifieras i 91-100% av proverna. Medianhalten var 21 ng/ml (16 μg/g kreatinin) för total As, 5,4 ng/ml (4,0 μg/g kreatinin) för summan av AsIII, AsV, MMA och DMA samt 11 ng/ml (8,5 μg/g kreatinin) för arsenobetain. Summan av AsIII, AsV, MMA och DMA låg på liknande nivå som i andra studier av barn och ungdomar. Pojkar hade signifikant högre urinhalter av arsenobetain än flickor, troligen på grund av att pojkar som deltog i Riksmaten ungdom 2016-17 konsumerade mer fisk än flickor. Det fanns inga tydliga skillnader i arsenikexponering mellan åldersgrupperna, men deltagare i årskurs 8 hade högre urinhalter av MMA än deltagare i gymnasiets årskurs 2

Contaminants in blood and urine from adolescents in Sweden : Results from the national dietary survey Riksmaten Adolescents 2016–17

I denna rapport sammanfattar vi biomonitoreringen av kemiska föroreningar i blod och urin från deltagarna i matvaneundersökningen Riksmaten ungdom 201617. Data från matvaneundersökningar som denna utgör en viktig vetenskaplig grund för Livsmedelsverkets risk- och nyttovärderingar. De är också viktiga när vi arbetar med riskhantering som kostråd, livsmedelskontroll och livsmedelslagstiftning. Riksmaten ungdom 201617 är en nationell tvärsnittsstudie där Livsmedelsverket undersökt matvanorna hos barn och ungdomar i årskurserna 5 och 8 samt andra året på gymnasiet (dvs. i åldrarna 1112, 1415 respektive 1718 år). Vi bjöd in representativa skolor över hela Sverige för att delta. Samtliga elever i en till två klasser vid de deltagande skolorna tillfrågades. I den här rapporten ingår den delgrupp av deltagare i Riksmaten ungdom 2016–17 där blod- och urinprover samlades in. Vi rekryterade elever från skolorna mellan september 2016 och maj 2017. Av de 259 skolor som blev inbjudna för blod- och urinprovtagning deltog 62 (24 procent). Vid dessa skolor deltog 1 305 av de 2 377 tillfrågade eleverna, varav 1 105 elever (46 procent) både gav fullständig information om matvanor och lämnade blod- och urinprover. 56 procent av deltagarna var flickor. 30 procent gick i årskurs 5, 37 procent i årskurs 8 och 32 procent i årskurs 2 på gymnasiet. Skolorna fanns i regionerna för Sveriges sju arbets- och miljömedicinska enheter (Göteborg, Linköping, Lund, Stockholm, Umeå, Uppsala och Örebro). I de blod- och urinprover som vi samlade in undersöktes ett brett spektrum av kemiska föroreningar: ämnen som inte längre är godkända för användning men ännu finns i miljön, ämnen som fortfarande produceras och används samt naturligt förekommande metaller. Vi analyserade följande grupper av substanser: klorerade och bromerade persistenta organiska föroreningar, högfluorerade ämnen, metaller, ftalatmetaboliter och flera fenolära ämnen (bl.a. bisfenoler och metaboliter av några bekämpningsmedel). Analysen av klorerade organiska föroreningar i serum visade att halterna var högst av DDT-metaboliten DDE, följt av PCB-153, hexaklorbensen (HCB), PCB-138 och PCB-180. Av den totala halten PCB utgjorde PCB-153, PCB-138 och PCB-180 ca 70 procent. Pojkarna i undersökningen hade högre halter än flickorna av de flesta klorerade föreningar. Halterna av DDE och vissa typer av PCB var dessutom högre bland de äldre deltagarna. Av de högfluorerade ämnena uppmätte vi högst serumhalter av PFOS och PFOA. De högsta halterna fanns hos ungdomar från Ronneby kommun (region Lund), där dricksvattnet tidigare varit kraftigt förorenat av dessa ämnen. Ämnena PFNA, PFHxS och PFOS fanns i högre halter hos pojkar än hos flickor. Nästan alla deltagare hade mätbara halter i blodet av kadmium, kvicksilver, bly, krom, mangan, kobolt och nickel. Aluminium kunde mätas i serum hos hälften av deltagarna. Hos pojkar fanns högre halter av kvicksilver och bly, medan halterna av kadmium var högre hos flickor. Hos 123 av deltagarna undersökte vi om det fanns arsenik i urinen. Alla prover hade mätbara halter av oorganisk arsenik eller minst en av de två analyserade organiska metaboliterna (monometylarsenik ochdimetylarsenik). Högst var halten av dimetylarsenik. Pojkar hade högre totalhalter av arsenik än flickor. Vi analyserade även en rad ftalatmetaboliter och fenolära ämnen i urin från deltagarna. Nästan alla (94100 procent) hade mätbara halter av ftalatmetaboliter och metaboliter till DiNCH (mjukgörare som används som ersättare till ftalater) i urinen. Högst halter fanns av monobutylftalat (MnBP), följt av monoetylftalat (MEP). Fenolära ämnen som kunde mätas i de flesta urinprover (mer än 90 procent) var bisfenol A och S, difenylfosfat (DPP, metabolit till ett fosforbaserat flamskyddsmedel), trikloropyridinol (metabolit till insekticiden klorpyrifos) och 3-fenoxybensoesyra (metabolit till pyretroider, en grupp bekämpningsmedel). Triklosan var mätbart hos 81 procent av deltagarna. Halterna av ftalatmetaboliter, DPP och bisfenol S var något högre hos flickor än hos pojkar. De ftalatmetaboliter och fenolära ämnen som undersöktes är kortlivade och försvinner snabbt ur kroppen. Att de ändå kunde mätas i urin tyder på en kontinuerlig exponering. Sammanfattningsvis kunde vi mäta föroreningar ur alla undersökta substansgrupper hos de flesta deltagare i Riksmaten ungdom 2016–17. Nivåerna var generellt jämförbara med nivåerna i andra studier och inom de intervall som kunde förväntas. Vi såg vissa köns- och åldersrelaterade skillnader i halter. Detta tyder på att det kan finnas skillnader i exponering, upptag eller eliminering mellan kön och åldersgrupper. Deltagarna var dock i olika tillväxtfaser och stadier av puberteten. Att göra en fullständig värdering av riskerna med de halter som uppmätts ligger utanför syftet med denna rapport. För några substanser finns det dock föreslagna halter i människokroppen under vilka det är osannolikt med negativa hälsoeffekter. Detta gäller vissa PCB-varianter, PFOS, PFOA, kvicksilver, bly, några ftalat- och DiNCH-metaboliter, bisfenol A samt triklosan. Våra resultat visar att de flesta svenska ungdomar har halter i sina kroppar som utifrån nuvarande kunskap sannolikt inte utgör någon risk för hälsan. Vissa individer hade emellertid högre halter av PFOS eller bly. Detta gällde framför allt bly. Där hade 7 procent av deltagarna blodhalter över Efsas referenspunkt för ökad risk för kronisk njursjukdom hos vuxna och 13 procent hade halter över referenspunkten för påverkan på hjärnans utveckling hos foster och små barn. Detta understryker vikten av att ytterligare minska exponeringen för bly från alla källor. Vi kan heller inte utesluta att det finns grupper i Sverige med högre exponering för kemiska föroreningar än deltagarna i Riksmaten ungdom 2016–17. Halterna av föroreningar varierade mycket mellan de som deltog i undersökningen. För att kunna bedöma och hantera risker är det viktigt att undersöka möjliga orsaker till dessa variationer. Därför kommer Livsmedelsverket att fortsätta utvärdera data från Riksmaten ungdom 2016–17. Detta kommer vi att göra genom att studera samband mellan halter av kemiska föroreningar och faktorer som kost, sociodemografi och livsstil.Dietary and biomonitoring data constitute an important scientific basis for risk and benefit assessments as well as for the development of risk management measures such as dietary advice, control programmes and food regulations. The present report summarises the results from biomonitoring of contaminants in blood and urine from participants in the dietary survey Riksmaten ungdom 2016–17 (Riksmaten Adolescents 2016–17). Riksmaten Adolescents 2016–17 is a nationally representative, cross-sectional, school-based dietary survey of children and adolescents in grades 5 and 8, and high school grade 2 (approximately 11–12, 14–15 and 17–18 years of age). Representative schools across Sweden were invited to take part in the study. At the participating schools, all students in one or two classes were invited. The study population of the present report was a subgroup of Riksmaten Adolescents 2016–17, from whom blood and urine were collected. Adolescents were recruited from schools between September 2016 and May 2017. Sixty-two (24%) of the 259 schools invited for blood and urine sampling participated. At these schools, 1,305 of the 2,377 invited students participated. Complete dietary information and valid blood and urine samples were available from 1,105 students (46%) and these were included in the analyses in this report. Fifty-six percent of the participants were girls. The distribution of participants between grades were as follows; grade 5: 30%, grade 8: 37%, and high school grade 2: 32%. The participating schools were distributed across Sweden’s seven Divisions of Occupational and Environmental Medicine (Gothenburg, Linköping, Lund, Stockholm, Umeå, Uppsala, and Örebro regions). A wide range of contaminants were investigated in the collected blood and urine samples: substances the use of which has been restricted or banned but which continue to persist in the environment; substances that continue to be legally produced and used; and naturally occurring toxic metals and trace elements. The analysed substance groups included chlorinated and brominated persistent organic pollutants, per- and polyfluoroalkyl substances (PFAS), metals and metalloids, phthalate metabolites and phenolic substances (e.g. bisphenols and metabolites of some pesticides). The chlorinated persistent organic pollutant with the highest concentration in serum was the DDT metabolite DDE, followed by PCB-153, hexachlorobenzene (HCB), PCB-138 and PCB-180. These three PCBs accounted for 70% of the total body burden of PCBs. Boys had higher serum concentrations of most chlorinated persistent organic pollutants than girls. Concentrations of DDE and some of the PCBs were higher in older age groups. PFOS and PFOA were the PFAS detected in the highest concentrations in serum. The highest concentrations of PFAS were found in individuals from Ronneby municipality (region Lund) where previously the drinking water has been contaminated with these compounds. Higher concentrations of PFNA, PFHxS and PFOS were observed in boys than girls. Almost all participants had detectable concentrations of cadmium, mercury, lead, chromium, manganese, cobalt and nickel in whole blood. Aluminium was detected in serum from half of the participants. Higher concentrations of mercury and lead were observed in boys than girls whereas blood concentrations of cadmium were higher in girls than boys. Urinary arsenic was measured in a subsample of 123 participants. All participants had quantifiable urinary levels of inorganic arsenic or at least one of its two analysed metabolites (monomethylarsonic acid and dimethylarsinic acid). The highest concentration was observed for dimethylarsinic acid. Boys had higher urine concentrations of total arsenic than girls. A number of phthalate metabolites and phenolic substances were analysed in urine from the participants. Almost all samples (94–100%) had measurable concentrations of phthalate metabolites and metabolites of the alternative plasticizer DiNCH. Of the phthalate metabolites, the highest concentrations were observed for mono-butyl phthalate (MnBP), followed by monoethyl phthalate (MEP). The phenolic substances detected in most samples (>90%) were bisphenol A and S, diphenyl phosphate (DPP, metabolite of a phosphorus-based flame retardant), trichloropyridinol (metabolite of the insecticide chlorpyrifos) and 3-phenoxybenzoic acid (metabolite of pyrethroids, a group of pesticides). Triclosan was detected in 81% of the samples. Concentrations of phthalate metabolites, DPP and bisphenol S were higher in girls than in boys. All the metabolites and phenolic substances analysed in urine are rapidly metabolised and excreted from the body. The detection of these substances thus suggests continuous exposure. In summary, contaminants from all investigated substance groups (i.e. chlorinated persistent organic pollutants, PFAS, metals and metalloids, and phthalates and phenolic compounds) could be quantified in samples from most participants in Riksmaten Adolescents 2016–17. However, the levels were generally comparable to levels found in other studies and within the expected ranges. Despite the heterogeneity of the study population, which represented a mix of individuals at different pubertal stages and growth phases, we found some gender- and age-related differences in contaminant concentrations. This suggests that there may be differences in exposure, uptake and/or elimination between genders and age groups. Full risk assessments of the observed concentrations were beyond the scope of this report. However, for some PCBs, PFOS, PFOA, mercury, lead, some phthalate and DiNCH metabolites, bisphenol A and triclosan, concentrations in the human body have been proposed below which it is unlikely that these contaminants cause adverse health effects. Our results show that, based on current knowledge, the levels of these contaminants in most Swedish adolescents are not a health concern. However, some participants exhibited a higher exposure to PFOS or lead. This was especially pronounced for lead, where 7% and 13% of the participants had blood concentrations above the EFSA reference points for increased risk of chronic kidney disease in adults and developmental neurotoxicity in small children, respectively. This underlines the importance of further reducing lead exposure from all sources, not only food. Moreover, we cannot exclude that there are Swedish populations with a higher exposure to contaminants than the populations covered by Riksmaten Adolescents 2016–17. There were substantial individual variations in the levels of the investigated contaminants among Swedish adolescents. For risk assessment and risk management purposes, it is important to explore possible causes of this variation. Therefore, the Swedish Food Agency will continue to evaluate data from Riksmaten Adolescents 2016–17 through further studies on the association between contaminant levels and factors such as diet, sociodemographics and lifestyle. 1

Concentrations of phthalates and phenolic substances in urine from first-time mothers in Uppsala, Sweden: temporal trends 2009- 2018

Sedan 1996 samlas blod- och modersmjölksprover regelbundet in från förstföderskor i Uppsala i den så kallade POPUP-studien. Sedan 2009 tas också ett urinprov. I denna rapport har tidstrender för ftalater och fenolära ämnen studerats i urinprov insamlade mellan 2009 och 2018. Ftalater och fenolära ämnen metaboliseras relativt snabbt i kroppen och för flertalet är det därför en metabolit till själva huvudsubstansen som har analyserats. Totalt sett analyserades tolv metaboliter till sex ftalater, en metabolit till en ersättningskemikalie till ftalater, metaboliter till tre fosforbaserade flamskyddsmedel, två pesticidmetaboliter samt åtta fenolära ämnen, bland annat triclosan, bisfenol A, S och F. Analyserna utfördes av Lunds universitet. Syftet var att studera tidstrender för dessa olika ämnen under perioden 2009-2018. Resultaten visade en nedåtgående tidstrend för de ftalater som håller på att fasas ut samtidigt som metaboliten för en ersättare till ftalaterna ökade. Det omdiskuterade ämnet bisfenol A och triclosan visade nedåtgående trender medan en ersättningssubstans till bisfenol A, bisfenol F, snarare hade en omvänd u-formad kurva. Även en metabolit till insekticiden klorpyrifos minskade under perioden, möjligen som en konsekvens av att strängare gränsvärden införts i EU. Analyser av urin gör det möjligt att studera hur befolkningens exponering för snabbmetaboliserande substanser ser ut. Genom att analysera prover över tid kan man studera hur befolkningens exponering förändras efter att åtgärder för att begränsa vissa kemikalier satts in samt hur exponeringen för nya ersättningskemikalier utvecklas

Maximum Tolerated Dose and Pharmacokinetics of Paclitaxel Micellar in Patients with Recurrent Malignant Solid Tumours : A Dose-Escalation Study

Introduction: A water-soluble Cremophor EL-free formulation of paclitaxel, in which retinoic acid derivates solubilize paclitaxel by forming micelles (paclitaxel micellar), was studied for the first time in man to establish the maximum tolerated dose (MTD) and to characterize the pharmacokinetics (PK). Methods: This was an open-label, one-arm, dose-escalating study in patients with advanced solid malignant tumours, for which no standard therapy was available or had failed. Paclitaxel micellar was given as 1-h intravenous infusion every 21 days for 3 cycles, mainly without premedication. Plasma samples were collected during 24 h at the first cycle and paclitaxel concentrations were assayed by high-performance liquid chromatography. PK was evaluated using a two-compartment model. Results: Thirty-four patients received paclitaxel micellar at doses ranging between 90 and 275 mg/m2. MTD was established as 250 mg/m2. Fatigue and neuropathy were the most frequent dose-limiting toxicities. No hypersensitivity reactions were observed. PK of paclitaxel was evaluated in 25 data sets. Paclitaxel micellar had a rapid initial distribution phase, mean half-life 0.55 h, estimated to be completed 3 h after dosing and a mean terminal half-life of 8.8 h. Mean clearance was 13.4 L/h/m2 with fivefold interindividual variability. The residual areas after 10 h and 24 h were 15.7 ± 8.6% and 5.7 ± 3.9% of the area under the plasma concentration–time curve to infinite time (AUCinf), respectively. Conclusion: No new side effects unknown for paclitaxel were observed. Maximum plasma concentration (Cmax) and AUCinf showed a tendency to increase linearly with dose within the 150–275 mg/m2dose range. The possibility to administer paclitaxel micellar without steroid premedication makes it an attractive candidate for further studies in combination with immunotherapy. Trial Registration: EudraCT no: 2004-001821-54