Eicosanoid Concentrations Research Articles

Pharmacokinetics of intravenous flumetasone and effects on plasma hydrocortisone concentrations and inflammatory mediators in the horse.

Flumetasone is a potent corticosteroid reportedly used in horses to decrease inflammation associated with strenuous exercise. There are currently no reports describing the use of this drug in horses. To describe the pharmacokinetics and effects on cortisol and eicosanoid concentrations, following administration of flumetasone to exercised horses. Parallel design. Twelve exercised horses received a single i.v. administration of 5 mg of flumetasone. Blood and urine samples were collected before and for 72 h post-drug administration for determination of flumetasone and cortisol concentrations. Whole blood samples were collected at various time and challenged with lipopolysaccharide, calcium ionophore or methanol to induce ex vivo synthesis of eicosanoids. Concentrations of flumetasone, cortisol and eicosanoids were measured using LC-MS/MS and pharmacokinetic/pharmacodynamic analysis performed. Flumetasone was detected for 23.5 ± 1.73 h in blood. The volume of distribution at steady state, systemic clearance and elimination half-life was 5.90 ± 0.200 L/kg, 30.7 ± 0.166 mL/min/kg and 4.84 ± 0.83 h respectively. Cortisol concentrations were still suppressed at last time point collected (72 h). For cortisol, Kin , Kout and the t1/2out were 30.3 ± 1.56 ng/mL × h, 0.331 ± 0.02 1/h and 2.1 h respectively. Stimulation with lipopolysaccharide resulted in a decrease in TXB2 , PGF2 , LTB4 , 15-HETE and 5-HETE for up to 72 h and PGE2 for 24 h post-flumetasone administration. Stimulation of whole blood with calcium ionophore resulted in a decrease in LTB4 for up to 6 h and 15-HETE at 8 h. Lack of sample collection for determination of biomarker concentrations beyond 72 h and the use of a single sample for determination of baseline cortisol concentrations. Flumetasone is rapidly cleared from blood following administration to horses. It is a potent anti-inflammatory with prolonged effects on production of cortisol and other inflammatory mediators.

Effect of psychiatric drugs on Daphnia magna oxylipin profiles

Neuro-active pharmaceuticals have been reported to act as endocrine disruptors enhancing reproduction in the model crustacean Daphnia manga at environmental concentrations of ng/L. Oxylipins and more specifically eicosanoids, which are lipid mediators formed from polyunsaturated fatty acids (PUFAs), are known to regulate reproduction together with other physiological processes in insects. In D. magna, the biosynthesis of eicosanoids and their putative role in the regulation of reproduction has been studied using transcriptomics, genomics and exposures to cyclooxygenase inhibitors. Quantification of eicosanoids and oxylipins derived from PUFAs upon exposure to pharmaceuticals is therefore crucial for a better understanding of the mode of action of neuro-active pharmaceuticals on aquatic invertebrates. The aim of this study was to investigate shifts in the oxylipin profile in D. magna adults upon exposure to environmental concentrations of the three psychiatric drugs, fluoxetine, diazepam and carbamazepine, with known effects of enhancing offspring production. Oxylipin profiles were determined in whole organism tissues using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Up to 28 different oxylipins belonging to arachidonic (AA), linoleic acid (LA), α-linoleic acid (α-LA) and eicosapentaenoic acid (EPA) pathways were detected and quantified in D. magna adults. Exposure to the selected psychiatric drugs showed that fluoxetine enhanced the accumulation of the cyclooxygenase (COX) product 12-hydroxyheptadecatrienoic acid (12-HHTrE), whereas diazepam increased the concentration of eicosanoids belonging to the lipoxygenase (LOX) and cytochrome P450 (CYP) pathways (HETEs, EpOMEs, HODEs, HOTrEs and HEPEs) from the AA, LA, αLA and EPA pathways. Carbamazepine had little effect and only one LA-derived compound from the LOX pathway (13-HODE) increased significantly. This means that despite having different modes of action in humans, fluoxetine and diazepam up-regulated eicosanoid pathways in D. magna, closely related to known biologically active products that regulate reproduction in insects.

Effects of medical ozone upon healthy equine joints: Clinical and laboratorial aspects.

ObjectiveThe aim of this study was to verify whether transient inflammatory reactions induced by intra-articular medicinal ozone administration affect joint components, by in vivo evaluation of inflammatory (prostaglandin E2, Substance P, Interleukin-6, Interleukine-1, Tumor Necrosis Factor), anti-inflammatory (Interleukin-10) and oxidative (superoxide dismutase activity and oxidative burst) biomarkers and extracellular matrix degradation products (chondroitin sulphate and hyaluronic acid) in synovial fluid.MethodsThe effects of medicinal ozone were analyzed at two ozone concentrations (groups A and B, 20 and 40 μg/ml, respectively), using oxygen-injected joints as controls (group C); each group received ten treatments (15 ml gas per treatment). Physical evaluation, evaluation of lameness, ultrasonography, and synovial fluid analysis were performed.ResultsAll joints presented mild and transient effusion throughout the study. Group B exhibited the highest lameness score on day 14 (P<0.05), detected by the lameness measurement system, probably because of the higher ozone concentration. All groups exhibited increased ultrasonography scores on day 14 (P < 0.05). Groups A and B exhibited increased proteins concentrations on day 21 (P<0.05). There was no change in hyaluronic acid concentration or the percentage of high-molecular weight hyaluronic acid throughout the experiment. Chondroitin sulfate concentrations decreased in group B, and did not change in group A and C, indicating that neither treatment provoked extracellular matrix catabolism. Cytokine and eicosanoid concentrations were not significantly changed.ConclusionsThe ozonetherapy did not cause significant inflammation process or cartilage degradation, therefore, ozonetherapy is safe at both evaluated doses.

Fish oil feeding attenuates neuroinflammatory gene expression without concomitant changes in brain eicosanoids and docosanoids in a mouse model of Alzheimer’s disease

BackgroundNeuroinflammation is a recognized hallmark of Alzheimer’s disease, along with accumulation of amyloid-β plaques, neurofibrillary tangles and synaptic loss. n-3 polyunsaturated fatty acids (PUFA) and molecules derived from them, including eicosapentaenoic acid-derived eicosanoids and docosahexaenoic acid-derived docosanoids, are known to have both anti-inflammatory and pro-resolving properties, while human observational data links consumption of these fatty acids to a decreased risk of Alzheimer’s disease. Few studies have examined the neuroinflammation-modulating effects of n-3 PUFA feeding in an Alzheimer’s disease-related model, and none have investigated whether these effects are mediated by changes in brain eicosanoids and docosanoids. Here, we use both a fat-1 transgenic mouse and a fish oil feeding model to study the impact of increasing tissue n-3 PUFA on neuroinflammation and the production of pro-inflammatory and pro-resolving lipid mediators. MethodsFat-1 mice, transgenic animals that can convert n-6 to n-3 PUFA, and their wildtype littermates were fed diets containing either fish oil (high n-3 PUFA) or safflower oil (negligible n-3 PUFA) from weaning to 12 weeks. Animals then underwent intracerebroventricular infusion of either amyloid-β 1-40 or a control peptide. Hippocampi were collected from non-surgery and surgery animals 10 days after infusion. Microarray was used to measure enrichment of inflammation-associated gene categories and expression of genes involved in the synthesis of lipid mediators. Results were validated by real-time PCR in a separate cohort of animals. Lipid mediators were measured via liquid chromatography tandem mass spectrometry. ResultsFat-1 and wildtype mice fed fish oil had higher total hippocampal DHA than wildtype mice fed the safflower oil diet. The safflower-fed mice, but not the fat-1 or fish oil-fed mice, had significantly increased expression in gene ontology categories associated with inflammation in response to amyloid-β infusion. These effects were independent of changes in the expression of genes involved in the synthesis of eicosanoids or docosanoids in any group. Gene expression was replicated upon validation in the wildtype safflower and fish oil-fed, but not the fat-1 mice. Protectin, maresin and D and E series resolvins were not detected in any sample. There were no major differences in levels of other eicosanoids or docosanoids between any of the groups in response to amyloid-β infusion. ConclusionsFish oil feeding decreases neuroinflammatory gene expression in response to amyloid-β. Neither amyloid-β infusion or increasing brain DHA affects the brain concentrations of specialized pro-resolving mediators in this model, or the concentrations of most other eicosanoids and docosanoids.

Multiplex quantitative analysis of eicosanoid mediators in human plasma and serum: Possible introduction into clinical testing

Eicosanoid mediators play important roles in maintaining the physiological and pathophysiological homeostasis in the body. Their measurements, however, are rarely performed in clinical practice. In the present study, we analyzed 30 varieties of eicosanoid mediators that were detectable in human plasma and serum collected from healthy donors, using liquid chromatography-tandem mass spectrometry from the viewpoint of the clinical application of the multiplex quantitation of eicosanoid mediators. Wider variety of eicosanoid mediators were detected in serum (27 out of 30) than in plasma (14 out of 30), since the serum was thought to contain lipid mediators released from activated platelets. Larger inter-individual variations were observed in the plasma and serum eicosanoid levels. On the other hand, the concentrations of eicosanoids were not affected by the platelet count but were affected by the concentration of arachidonic acid (AA) within the reference interval (17.4-40.5×1010/L). When serum samples from patients with hematological disorders were analyzed, the concentrations of AA were positively correlated with the platelet count. When the patients underwent ASA therapy, a marked decrease in the concentrations of thromboxane B2 (TXB2) and 12-hydroxyl-heptadecatrienoic acid (12-HHT) was observed. Considering the availability of serum samples in clinical settings, the serum analysis of eicosanoids may be clinically useful.

The effect of allergen-induced bronchoconstriction on concentration of 5-oxo-ETE in exhaled breath condensate of house dust mite-allergic patients.

Arachidonic acid metabolites regulate several aspects of airway function including inflammation, muscle contraction and mucous secretion. The aim of this study was to evaluate concentration of selected 5-lipoxygenase- and cyclooxygenase-derived eicosanoids in exhaled breath condensate (EBC) during allergen-induced bronchoconstriction. The study was performed on 24 allergic rhinitis/asthma patients sensitized to a house dust mite (HDM) Dermatophagoides pteronyssinus (Dp) and 13 healthy controls (HCs). Bronchial challenge with Dp extract was performed only in the allergic patients. EBC samples were collected before (T0 ) and during Dp-induced bronchoconstriction (TEAR ). Eicosanoid concentration was measured using HPLC-tandem mass spectrometry. Significant bronchoconstriction after Dp challenge was demonstrated in 15 patients (Rs), while in 9 patients (NRs) no asthmatic response could be detected. At T0 the most abundant eicosanoids in EBC of HDM-allergic patients were LTB4 and 5-oxo-ETE, while in HCs EBC concentration of LTB4 was significantly greater than that of 5-oxo-ETE. Allergen challenge resulted in significant increase in EBC concentration of 5-oxo-ETE, LTD4 and 8-iso-PGE2 only in Rs. At TEAR , the relative change of 5-oxo-ETE concentration in EBC correlated with decrease of peripheral blood eosinophilia (R=-0.774; P=.0012). Moreover, the relative increase of 5-oxo-ETE in EBC at TEAR significantly correlated with the severity of the subsequent late asthmatic response (R=0.683, P=.007). Our study demonstrates significant up-regulation of 5-oxo-ETE synthesis in HDM-allergic patients and indicates possible involvement of that mediator in the pathogenesis of allergic asthma.

Impact of conjugated linoleic acid administered to rats prior and after carcinogenic agent on arachidonic and linoleic acid metabolites in serum and tumors

The objective of the study was to assess the influence of conjugated linoleic acid (CLA) daily supplementation prior and after carcinogenic agent on the concentrations of eicosanoids – metabolites of arachidonic acid (15-, 12- or 5-hydroxyeicosatetraenoic acids (15-, 12-, 5-HETE), prostaglandin E2 (PGE2)) and linoleic acid (13- or 9-hydroxyoctadecadienoic acids (13-, 9-HODE)) in rat serum and 7,12-dimethylbenz[a]anthracene (DMBA)-induced tumors. Female rats were randomised into six groups, receiving 1% or 2% Bio-C.L.A or plant oil since the 37th day of life throughout the whole experiment. Some rats (50-day-old) were administered DMBA to induce tumors. Eicosanoids were analyzed with LC-MS/MS. The study indicated that CLA supplemented daily to rats prior and after carcinogen administration affected concentrations of arachidonic and linoleic acid metabolites in rat serum and induced tumors. However, ratios of eicosanoids exerting opposite activities (e.g. 12-HETE/15-HETE) appear to act as more precise factors reflecting pathological changes in an organism than individual compounds.

Machine-Learned Data Structures of Lipid Marker Serum Concentrations in Multiple Sclerosis Patients Differ from Those in Healthy Subjects.

Lipid signaling has been suggested to be a major pathophysiological mechanism of multiple sclerosis (MS). With the increasing knowledge about lipid signaling, acquired data become increasingly complex making bioinformatics necessary in lipid research. We used unsupervised machine-learning to analyze lipid marker serum concentrations, pursuing the hypothesis that for the most relevant markers the emerging data structures will coincide with the diagnosis of MS. Machine learning was implemented as emergent self-organizing feature maps (ESOM) combined with the U*-matrix visualization technique. The data space consisted of serum concentrations of three main classes of lipid markers comprising eicosanoids (d = 11 markers), ceramides (d = 10), and lyosophosphatidic acids (d = 6). They were analyzed in cohorts of MS patients (n = 102) and healthy subjects (n = 301). Clear data structures in the high-dimensional data space were observed in eicosanoid and ceramides serum concentrations whereas no clear structure could be found in lysophosphatidic acid concentrations. With ceramide concentrations, the structures that had emerged from unsupervised machine-learning almost completely overlapped with the known grouping of MS patients versus healthy subjects. This was only partly provided by eicosanoid serum concentrations. Thus, unsupervised machine-learning identified distinct data structures of bioactive lipid serum concentrations. These structures could be superimposed with the known grouping of MS patients versus healthy subjects, which was almost completely possible with ceramides. Therefore, based on the present analysis, ceramides are first-line candidates for further exploration as drug-gable targets or biomarkers in MS.

Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate

BackgroundResearchers investigating lung diseases, such as asthma, have questioned whether certain compounds previously reported in exhaled breath condensate (EBC) originate from saliva contamination. Moreover, despite its increasing use in ‘omics profiling studies, the constituents of EBC remain largely uncharacterized. The present study aims to define the usefulness of EBC in investigating lung disease by comparing EBC, saliva, and saliva-contaminated EBC using targeted and untargeted mass spectrometry and the potential of metabolite loss from adsorption to EBC sample collection tubes.MethodsLiquid chromatography mass spectrometry (LC-MS) was used to analyze samples from 133 individuals from three different cohorts. Levels of amino acids and eicosanoids, two classes of molecules previously reported in EBC and saliva, were measured using targeted LC-MS. Cohort 1 was used to examine contamination of EBC by saliva. Samples from Cohort 1 consisted of clean EBC, saliva-contaminated EBC, and clean saliva from 13 healthy volunteers; samples were analyzed using untargeted LC-MS. Cohort 2 was used to compare eicosanoid levels from matched EBC and saliva collected from 107 asthmatic subjects. Samples were analyzed using both targeted and untargeted LC-MS. Cohort 3 samples consisted of clean-EBC collected from 13 subjects, including smokers and non-smokers, and were used to independently confirm findings; samples were analyzed using targeted LC-MS, untargeted LC-MS, and proteomics. In addition to human samples, an in-house developed nebulizing system was used to determine the potential for EBC samples to be contaminated by saliva.ResultsOut of the 400 metabolites detected in both EBC and saliva, 77 were specific to EBC; however, EBC samples were concentrated 20-fold to achieve this level of sensitivity. Amino acid concentrations ranged from 196 pg/mL – 4 μg/mL (clean EBC), 1.98 ng/mL – 6 μg/mL (saliva-contaminated EBC), and 13.84 ng/mL – 1256 mg/mL (saliva). Eicosanoid concentration ranges were an order of magnitude lower; 10 pg/mL – 76.5 ng/mL (clean EBC), 10 pg/mL – 898 ng/mL (saliva-contaminated EBC), and 2.54 ng/mL – 272.9 mg/mL (saliva). Although the sample size of the replication cohort (Cohort 3) did not allow for statistical comparisons, two proteins and 19 eicosanoids were detected in smoker vs. non-smoker clean-EBC.ConclusionsWe conclude that metabolites are present and detectable in EBC using LC-MS; however, a large starting volume of sample is required.

Effects of Leukoreduction and Storage on Erythrocyte Phosphatidylserine Expression and Eicosanoid Concentrations in Units of Canine Packed Red Blood Cells.

BackgroundStorage of canine packed red blood cells (pRBCs) can increase erythrocyte phosphatidylserine (PS) expression and eicosanoid concentrations.Hypothesis/ObjectivesTo determine the effects of leukoreduction on erythrocyte PS expression and eicosanoid concentrations in stored units of canine pRBCs. Our hypothesis was that leukoreduction would decrease PS expression and eicosanoid concentrations.AnimalsEight healthy dogs.MethodsIn a cross‐over study, units of whole blood were leukoreduced (LR) or non‐LR and stored (10 and 21 days) as pRBCs. Samples were collected at donation, and before and after a simulated transfusion. PS expression was measured by flow cytometry, and concentrations of arachidonic acid (AA), prostaglandin F2α (PGF 2α), prostaglandin E2 (PGE 2), prostaglandin D2 (PGD 2), thromboxane B2 (TXB 2), 6‐keto‐prostaglandin F1α (6‐keto‐PGF 1α), and leukotriene B4 (LTB 4) were quantified by liquid chromatography–mass spectrometry.ResultsThere was no change in PS expression during leukoreduction, storage, and simulated transfusion for non‐LR and LR units. Immediately after leukoreduction, there was a significant increase in TXB 2 and PGF 2α concentrations, but during storage, these eicosanoids decreased to non‐LR concentrations. In both LR and non‐LR units, 6‐keto‐PGF 1α concentrations increased during storage and simulated transfusion, but there was no difference between unit type. There was no difference in AA, LTB 4, PGE 2, and PGD 2 concentrations between unit types.Conclusions and Clinical ImportanceLeukoreduction, storage, and simulated transfusion do not alter erythrocyte PS expression. Leukoreduction causes an immediate increase in concentrations of TXB 2 and PGF 2α, but concentrations decrease to non‐LR concentrations with storage. Leukoreduction does not decrease the accumulation of 6‐keto‐PGF 1α during storage.

Evaluation of eicosanoid concentrations in stored units of canine packed red blood cells

OBJECTIVE To evaluate eicosanoid concentrations in freshly prepared canine packed RBCs (PRBCs) and to assess changes in eicosanoid concentrations in PRBC units over time during storage and under transfusion conditions. DESIGN Prospective study. SAMPLE 25 plasma samples from 14 healthy Greyhounds. PROCEDURES Plasma samples were obtained during PRBC preparation (donation samples), and the PRBC units were then stored at 4°C until used for transfusion (≤ 21 days later; n = 17) or mock transfusion if expired (22 to 24 days later; 8). Immediately prior to use, 100 mL of saline (0.9% NaCl) solution was added to each unit and a pretransfusion sample was collected. A posttransfusion sample was collected after transfusion or mock transfusion. Concentrations of arachidonic acid, prostaglandin (PG) F2α, PGE2, PGD2, thromboxane B2, 6-keto-PGF1α, and leukotriene B4 were measured by liquid chromatography-mass spectrometry and analyzed statistically. RESULTS Median arachidonic acid concentration was significantly decreased in posttransfusion samples, compared with the concentration in donation samples. Median PGF2α, 6-keto-PGF1α, and leukotriene B4 concentrations were significantly increased in pretransfusion samples, compared with those in donation samples. Median PGF2α, thromboxane B2, and 6-keto-PGF1α concentrations were significantly increased in posttransfusion samples, compared with those in pretransfusion samples. Duration of PRBC storage had significant associations with pretransfusion and posttransfusion arachidonic acid and thromboxane B2 concentrations. CONCLUSIONS AND CLINICAL RELEVANCE Concentrations of several proinflammatory eicosanoids increased in PRBC units during storage, transfusion, or both. Accumulation of these products could potentially contribute to adverse transfusion reactions, and investigation of the potential association between eicosanoid concentrations in PRBCs and the incidence of transfusion reactions in dogs is warranted.

Omega-3 fatty acids, inflammatory status and biochemical markers of patients with systemic lupus erythematosus: a pilot study

BackgroundStudies have shown that omega-3 fatty acids reduce the concentrations of eicosanoids, cytokines, chemokines, C-reactive protein (CRP) and other inflammatory mediators. ObjectiveTo investigate the effects of omega-3 fatty acids on circulating levels of inflammatory mediators and biochemical markers in women with systemic lupus erythematosus (SLE). MethodsExperimental clinical study (clinical trial: NCT02524795); 49 women with SLE (ACR1982/1997) were randomized: 22 to the omega-3 group (daily intake of 1080mg EPA+200mg DHA, for 12 weeks) and 27 to the control group. The inflammatory mediators and biochemical markers at T0 and T1 in omega-3 group were compared using Wilcoxon test. U-Mann–Whitney test was used to compare variations of measured variables [ΔV=pre-treatment (T0)−post-treatment (T1) concentrations] between groups. p<0.05 was considered significant. ResultsThe median (interquartile range – IQR) of age was 37 (29–48) years old, of disease duration was 7 (4–13) years, and of SLEDAI-2K was 1 (0–2). The median (IQR) of variation in CRP levels between the two groups showed a decrease in omega-3 group while there was an increase in control group (p=0.008). The serum concentrations of IL-6 and IL-10, leptin and adiponectin did not change after a 12 week treatment. ConclusionsSupplementation with omega-3 had no impact on serum concentrations of IL-6, IL-10, leptin and adiponectin in women with SLE and low disease activity. There was a significant decrease of CRP levels as well as evidence that omega-3 may impact total and LDL-cholesterol.

Ácidos graxos ômega‐3, estado inflamatório e marcadores bioquímicos de pacientes com lúpus eritematoso sistêmico: estudo piloto

BackgroundStudies have shown that omega‐3 fatty acids reduce the concentrations of eicosanoids, cytokines, chemokines, C‐reactive protein (CRP) and other inflammatory mediators. ObjectiveTo investigate the effects of omega‐3 fatty acids on circulating levels of inflammatory mediators and biochemical markers in women with systemic lupus erythematosus (SLE). MethodsExperimental clinical study (clinical trial: NCT02524795); 49 women with SLE (ACR1982/1997) were randomized: 22 to the omega‐3 group (daily intake of 1080mg EPA+200mg DHA, for 12 weeks) and 27 to the control group. The inflammatory mediators and biochemical markers at T0 and T1 in omega‐3 group were compared using Wilcoxon test. U‐Mann‐Whitney test was used to compare variations of measured variables [ΔV=pre‐treatment (T0) minus post‐treatment (T1) concentrations] between groups. p<0.05 was considered significant. ResultsThe median (interquartile range ‐ IQR) of age was 37 (29‐48) years old, of disease duration was 7 (4‐13) years, and of SLEDAI‐2K was 1 (0‐2). The median (IQR) of variation in CRP levels between the two groups showed a decrease in omega‐3 group while there was an increase in control group (p=0.008). The serum concentrations of IL‐6 and IL‐10, leptin and adiponectin did not change after a 12 week treatment. ConclusionsSupplementation with omega‐3 had no impact on serum concentrations of IL‐6, IL‐10, leptin and adiponectin in women with SLE and low disease activity. There was a significant decrease of CRP levels as well as evidence that omega‐3 may impact total and LDL‐cholesterol

Thrombophilia and Thrombotic Disorders in Newborns

incidence in children is estimated at about 0.7 cases of venous thrombosis per 100,000 population, 1.0 of stroke and 0.1 of myocardial infarction, while in adults the incidence reported is 74.2, 45.4 and 175.6 cases respectively. The lower incidence of venous thromboembolic disease (VTE) in children compared to adults is mainly due to the integrity of the vessels and the enhanced anticoagulant activity of the endothelium, and additionally to the reduced capacity of thrombin generation, inactivation of thrombin by a2-macroglobulin, the increased concentration of which, counterbalances the physiological deficiency of antithrombin and protein C / S, to the decreased levels of tissue factor in cord blood and concentration of eicosanoids and proteoglycans, depending on age [2,3]. The incidence of thromboembolic events (VTE) is higher in infants than in older children. About 10% of VTE occur in the first four weeks of life, frequently in critically ill neonates (2.4 symptomatic VTE cases per 1000 admissions in Neonatal Intensive Care Units-NICU). The incidence of symptomatic venous thrombosis (VTE) in newborns is 5.1/100,000 live births [4]. Although it is considered that the thromboembolic events are under diagnosed, newborns are nearly 40 times more likely to suffer a VTE than during the entire childhood, but certainly the above probability is significantly less than that referred in adults [5,6]. The hemostatic system both in preterm and full-term newborns differs significantly from that of older children and much more than that of adults, however, normally in newborns there is a balance in between bleeding or thrombosis diathesis. The increased incidence of VTE in critically ill infants is attributed to the derangement of equilibrium of the hemostatic mechanism, because of the lower concentrations of natural inhibitors and impaired fibrinolytic activity and also due to elevated levels of factor von Willebrand [7]. The combination of the above with both the high viscosity of blood due to the high hematocrit and the small vascular diameter of newborns results in low capillary flow, especially on coexistence of dehydration or hypercoagulable state because of infection or prematurity. Furthermore, the use of venous or arterial catheters is one of the major risk factors for thrombosis in neonatal period. Nowadays, 15% of infants in NICU and at least 50% of preterm infants of birth weight 90% of cases) or congenital risk factors. For the manifestation of VTE in neonates, several coincidental risk factors (variety of underlying diseases or triggering events of the child or maternal) are required. Studies so far, have not revealed thrombophilic factors contributing to the occurrence of asymptomatic or symptomatic VTE [8]. Genetic factors are related to the risk of VTE in children and newborns but no causal relationship is established for most of them [9]. The frequency of presence of thrombophilic factors in children with VTE is reported 13-79% in several studies. The huge discrepancy is attributed mainly to variation in the design of the trials, the difficulty in the definition of related prothrombotic disorders, and also to the small number and different patient populations. Regarding thromboembolic events of central venous system (CNS) [10], and although the high frequency of idiopathic arterial ischemic events (AIS) [11], the prevalence of thrombophilic factors varies in different studies from 20 to 50%, while increased appears the possibility of the presence of congenital thrombophilia in neonates with renal, hepatic or portal vein thrombosis [12]. Furthermore, maternal thrombophilia has been associated with the occurrence of perinatal AIS, venous sinus thrombosis or renal vein thrombosis [13]. However, in neonates there is scepticism about the interpretation of results of the various studies due to the complexity of the mechanism of hemostasis and the interaction of several acquired factors. Thrombosis in neonates is generally a multifactorial process [14]. Future studies may clarify both causative background and therapeutic choices in this group of patients.

Effect of Marine-Derived n-3 Polyunsaturated Fatty Acids on Major Eicosanoids: A Systematic Review and Meta-Analysis from 18 Randomized Controlled Trials.

BackgroundMarine-derived n-3 polyunsaturated fatty acids (PUFA) may have a beneficial effect on inflammation via lowering pro-inflammatory eicosanoid concentrations. We aimed to assess the effect of marine-derived n-3 PUFA on prostaglandin E2 (PGE2), thromboxane B2 (TXB2), and leukotriene B4 (LTB4) through systematic review and meta-analysis of randomized controlled trials.Method and FindingsA structured search strategy on PubMed, Web of Science and Cochrane up to November 2015 was undertaken in this meta-analysis. Standard mean difference was used to calculate the effect size of marine-derived n-3 PUFA on PGE2, TXB2 and LTB4 in a random-effect model. A total of 18 RCTs with 826 subjects were included in this systematic review and meta-analysis. Supplementation of marine-derived n-3 PUFA significantly decreased concentrations of TXB2 in serum/plasma in subjects with high risk of cardiovascular diseases (SMD:-1.26; 95% CI: -1.65, -0.86) and LTB4 in neutrophils in unhealthy subjects (subjects with non-autoimmune chronic diseases or auto-immune diseases) (SMD:-0.59: 95% CI: -1.02, -0.16). Subgroup analyses showed a significant reduction of LTB4 in subjects with rheumatoid arthritis (SMD: -0.83; 95% CI: -1.37, -0.29), but not in non-autoimmune chronic disease patients (SMD: -0.33; 95% CI: -0.97, 0.31). No significant publication bias was shown in the meta-analysis.ConclusionsMarine-derived n-3 PUFA had a beneficial effect on reducing the concentration of TXB2 in blood of subjects with high risk of CVD as well as LTB4 in neutrophils in unhealthy subjects, and that subjects with RA showed lower LTB4 content with supplementation of marine-derived n-3 PUFA.

Plasma Vasoprotective Eicosanoid Concentrations in Healthy Greyhounds and Non-Greyhound Dogs.

BackgroundHypertension and albuminuria often coexist in Greyhounds, suggesting generalized vascular dysfunction that could contribute to the development of a variety of diseases in this breed. Eicosanoid metabolites of arachidonic acid (AA) mediate endothelial function, vascular reactivity, and proteinuria in humans and in rodent models.HypothesisThe eicosanoid profile of Greyhounds is shifted toward metabolites that promote vascular dysfunction, hypertension, and proteinuria.AnimalsHealthy Greyhounds (n = 20) and non‐Greyhound (n = 20) dogs that were consecutively enrolled in a blood donor program.MethodsProspective study. Plasma eicosanoid metabolites were assayed by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI/MS) and compared to systolic blood pressure (SP) measurements and urine albumin concentration.ResultsIsomers of hydroxyeicosatetraenoic acid (HETE) were higher in Greyhounds than non‐Greyhounds (median, range in pmol/mL: 5(S)HETE 19.82, 8.55–32.95 versus 13.54, 4.33–26.27, P = .033; 8(S)HETE 9.39, 3.28–19.84 versus 5.80, 2.25–17.66, P = .002; 9(S)HETE 9.46, 2.43–13.79 versus 5.82, 1.50–17.16, P = .026; 12(S)HETE 10.17, 3.81–40.06 versus 7.24, 2.9–16.16, P = .022). Dihydroxyeicosatrienoic acid (DHET) isomers also were higher in Greyhounds compared to non‐Greyhounds (mean ± SD in pmol/mL: 8,9DHET 5.78 ± 2.13 versus 4.03 ± 1.36, P = .004; 11,12DHET 11.98 ± 2.86 versus 8.90 ± 3.48, P = .004; 14,15DHET 7.23 ± 2.19 versus 5.76 ± 1.87, P = .028). Albuminuria correlated with total DHET (rs = 0.46, P = .003). SP was positively correlated with 11,12EET (rs = 0.42, P = .006) and 20(S)HETE (rs = 0.38, P = .017). SP and 8,9EET were inversely correlated (rs = −0.49, P = .001).Conclusions and Clinical ImportancePlasma eicosanoid profile in Greyhounds was consistent with activation of metabolic pathways known to promote vascular dysfunction and might contribute to higher blood pressures and albuminuria. Inhibition of these eicosanoid pathways should be evaluated as therapeutic targets in Greyhounds.

In vivo solid-phase microextraction liquid chromatography–tandem mass spectrometry for monitoring blood eicosanoids time profile after lipopolysaccharide-induced inflammation in Sprague-Dawley rats

A fast and non-lethal in vivo solid-phase microextraction (SPME) sampling method for rat blood coupled to liquid chromatography and tandem mass spectrometry (LC–MS/MS) was developed for monitoring rapid changes in concentrations of eicosanoids – lipid mediators involved in the development of inflammatory conditions – using diffusion-based calibration. Sampling rates of target eicosanoids were pre-determined under laboratory conditions with a precision of ≤10%, and directly used for quantification of analyte concentrations in blood after lipopolysaccharide-induced inflammation in Sprague-Dawley rats. Results showed significant changes in unbound plasma concentrations of arachidonic acid (AA) and 12-hydroxyeicosatetraenoic acid (12-HETE) in response to the treatment. Next, performance of the proposed method was compared with protein precipitation (PP) of plasma, a conventional sample preparation technique. Finally, percentages of plasma protein binding (PPB) of specific eicosanoids were determined. PPB of target eicosanoids was in agreement with literature values, ranging from 99.3 to 99.9% for 12-HETE and DHA, respectively. We envision that the proposed method is a particularly suitable alternative to lethal sampling and current methods based on sample depletion in animal studies for accurate monitoring of rapid changes in blood concentrations of small molecules.

A randomized controlled trial of green tea catechins in protection against ultraviolet radiation–induced cutaneous inflammation

Background: Safe systemic protection from the health hazards of ultraviolet radiation (UVR) in sunlight is desirable. Green tea is consumed globally and is reported to have anti-inflammatory properties, which may be mediated through the impact on cyclooxygenase and lipoxygenase pathways. Recent data suggest that green tea catechins (GTCs) reduce acute UVR effects, but human trials examining their photoprotective potential are scarce.Objective: We performed a double-blind, randomized, placebo-controlled trial to examine whether GTCs protect against clinical, histologic, and biochemical indicators of UVR-induced inflammation.Design: Healthy adults (aged 18–65 y, phototypes I–II) were randomly allocated to 1350 mg encapsulated green tea extract (540 mg GTC) with 50 mg vitamin C or placebo twice daily for 3 mo. Impact on skin erythema, dermal leukocytic infiltration, and concentrations of proinflammatory eicosanoids was assessed after solar-simulated UVR challenge, and subject compliance was determined through assay of urinary GTC metabolite epigallocatechin glucuronide.Results: Volunteers were assigned to the active (n = 25) or the placebo (n = 25) group. After supplementation, median (IQR) sunburn threshold (minimal erythema dose) was 28 (20–28) and 20 (20–28) mJ/cm2 in the active and placebo groups, respectively (nonsignificant), with no difference in AUC analysis for measured erythema index after a geometric series of 10 UVR doses. Skin immunohistochemistry showed increased neutrophil and CD3+ T-lymphocyte numbers post-UVR in both groups (P < 0.01) with no statistically significant differences between groups after supplementation. Cyclooxygenase and lipoxygenase metabolites prostaglandin E2 (vasodilator) and 12-hydroxyeicosatetraenoicacid (chemoattractant), respectively, increased after UVR (P < 0.05), with no differences between supplementation groups.Conclusion: Oral GTC (1080 mg/d) with vitamin C over 3 mo did not significantly reduce skin erythema, leukocyte infiltration, or eicosanoid response to UVR inflammatory challenge. This trial was registered at clinicaltrials.gov as NCT01032031.

"A randomized, double-blind study of the effects of omega-3 fatty acids (Omegaven) on outcome after major liver resection".

BackgroundThe body is dependent on the exogenous supply of omega-3 polyunsaturated fatty acids (n3-PUFA). These essential fatty acids are key players in regulating metabolic signaling but also exert anti-inflammatory and anti-carcinogenic properties. The liver is a major metabolic organ involved in fatty acid metabolism. Under experimental conditions, n3-PUFA exert beneficial effect on hepatic steatosis, regeneration and inflammatory insults such as ischemic injury after surgery. Some of these effects have also been observed in human subjects. However, it is unclear whether perioperative administration of n3-PUFA is sufficient to protect the liver from ischemic injury. Therefore, we designed a randomized controlled trial (RCT) assessing n3-PUFA (pre-) conditioning strategies in patients scheduled for liver surgery.Methods/DesignThe Omegaven™ trial is a multi-centric, double-blind, randomized, placebo- controlled trial applying two single doses of Omegaven™ or placebo on 258 patients undergoing major liver resection. Primary endpoints are morbidity and mortality one month after hospital discharge, defined by the Clavien- Dindo classification of surgical complications (Ann Surg 240(2):205–13, 2004) as well as the Comprehensive Complication Index (CCI) (Ann Surg 258(1):1–7, 2013). Secondary outcome variables include length of Intensive Care Unit (ICU) and hospital stay, postoperative liver function tests, fatty acid and eicosanoid concentration, inflammatory markers in serum and in liver tissue. An interim analysis is scheduled after the first 30 patients per randomization group.DiscussionLong-term administration of n3-PUFA have a beneficial effect on metabolism and hepatic injury. Patients often require surgery without much delay, thus long-term n3-PUFA uptake is not possible. Also, lack of compliance may lead to incomplete n3-PUFA substitution. Hence, perioperative Omegaven™ may provide an easy and controllable way to ensure hepaative application of tic protection.Trial registrationClinicalTrial.gov: ID: NCT01884948, registered June 14, 2013; Institution Ethical Board Approval: KEK-ZH-Nr. 2010–0038; Swissmedic Notification: 2012DR3215.

Transient postnatal fluoxetine decreases brain concentrations of 20-HETE and 15-epi-LXA4, arachidonic acid metabolites in adult mice

BackgroundTransient postnatal exposure of rodents to the selective serotonin (5-HT) reuptake inhibitor (SSRI) fluoxetine alters behavior and brain 5-HT neurotransmission during adulthood, and also reduces brain arachidonic (ARA) metabolic consumption and protein level of the ARA metabolizing enzyme, cytochrome P4504A (CYP4A). HypothesisBrain 20-hydroxyeicosatetraenoic acid (20-HETE), converted by CYP4A from ARA, will be reduced in adult mice treated transiently and postnatally with fluoxetine. MethodsMale mice pups were injected i.p. daily with fluoxetine (10mg/kg) or saline during P4–P21. At P90 their brain was high-energy microwaved and analyzed for 20-HETE and six other ARA metabolites by enzyme immunoassay. ResultsPostnatal fluoxetine vs. saline significantly decreased brain concentrations of 20-HETE (−70.3%) and 15-epi-lipoxin A4 (−60%) in adult mice, but did not change other eicosanoid concentrations. ConclusionsBehavioral changes in adult mice treated postnatally with fluoxetine may be related to reduced brain ARA metabolism involving CYP4A and 20-HETE formation.