Unravelling the immunomodulatory role of apple phenolic rich extracts on human THP-1- derived macrophages using multiplatform metabolomics.

Apples represent a significant source of dietary phenolic compounds with evidenced anti-inflammatory and immunomodulatory activities. Nevertheless, the effect of the whole apple matrix on human macrophages is unknown. In this context, our study attempts to evaluate the effect of apple-derived phenolic compounds-rich extracts (pulp, peel and leaf) on IL-1ß production in THP-1-differentiated macrophages and derived metabolic alterations through untargeted metabolomics. Our results have showed that apple pulp treatment inhibited the release of the pro-inflammatory cytokine IL-1ß induced by LPS in THP-1 macrophages by ELISA analysis. Metabolomics demonstrate that different proportions of phenolic compounds led to differential alterations in the metabolism of THP-1 macrophages. Indeed, apple extracts promoted alterations in lipid, carbohydrate, amino acid and vitamins as well as cofactors metabolism. Specifically, leaf extracts were characterized by alteration of galactose metabolism while the extracts derived from the fruit showed predominant alterations in lipids metabolism. All extracts mimicked the response observed under normal conditions in LPS-stimulated macrophages, inhibiting LPS response. Thus, the phenolic enriched extracts from apples will be a good source of natural compounds with a beneficial effect against inflammation, and they may be applied as a food supplement and/or functional ingredient for the treatment of inflammatory diseases.

Applicability of an In-Vitro Digestion Model to Assess the Bioaccessibility of Phenolic Compounds from Olive-Related Products.

The Mediterranean diet includes virgin olive oil (VOO) as the main fat and olives as snacks. In addition to providing nutritional and organoleptic properties, VOO and the fruits (olives) contain an extensive number of bioactive compounds, mainly phenolic compounds, which are considered to be powerful antioxidants. Furthermore, olive byproducts, such as olive leaves, olive pomace, and olive mill wastewater, considered also as rich sources of phenolic compounds, are now valorized due to being mainly applied in the pharmaceutical and nutraceutical industries. The digestive system must physically and chemically break down these ingested olive-related products to release their phenolic compounds, which will be further metabolized to be used by the human organism. The first purpose of this review is to provide an overview of the current status of in-vitro static digestion models for olive-related products. In this sense, the in-vitro gastrointestinal digestion methods are widely used with the following aims: (i) to study how phenolic compounds are released from their matrices and to identify structural changes of phenolic compounds after the digestion of olive fruits and oils and (ii) to support the functional value of olive leaves and byproducts generated in the olive industry by assessing their health properties before and after the gastrointestinal process. The second purpose of this review is to survey and discuss all the results available to date.

Optimization of a new selective pressurized liquid extraction methodology for determining organic pollutants in wild boar livers.

In this study, a new selective pressurised liquid extraction (SPLE) methodology was optimised for determining about 70 organic pollutants (OPs) including organochlorine (OCPs), organophosphate (OPPs) and pyrethroid (PYRs) pesticides, polychlorinated biphenyls (PCBs), polybromodiphenyl ethers (PBDEs), as well as, polycyclic aromatic hydrocarbons (PAHs) in wild boar liver samples considering the temperature, pressure and time of contact between the solvent and the matrix as influential variables. Clean-up of extracts was performed by solid-phase extraction (SPE) using EZ-POP cartridges. Detection of OPs was carried out by gas chromatography (GC) coupled to tandem mass spectrometry (QqQ-MS/MS). This new approach offers:•A new non-time consuming SPLE methodology for determining about 70 OPs in wild boar.•Recoveries achieved ranged between 74 to 119 % with RSD less than 20 %.•Detection and quantification limits in the low to mid pg/g range.

Metabolomics Insights of the Immunomodulatory Activities of Phlorizin and Phloretin on Human THP-1 Macrophages.

Dihydrochalcones, phlorizin (PZ) and its aglycone phloretin (PT), have evidenced immunomodulatory effects through several mechanisms. However, the differential metabolic signatures that lead to these properties are largely unknown. Since macrophages play an important role in the immune response, our study aimed to characterise human THP-1 macrophages under PZ and PT exposure. A multiplatform-based untargeted metabolomics approach was used to reveal metabolites associated with the anti-inflammatory mechanisms triggered by the dihydrochalcones in LPS-stimulated macrophages, for the first time. Results showed differential phenotypic response in macrophages for all treatments. Dihydrochalcone treatment in LPS-stimulated macrophages mimics the response under normal conditions, suggesting inhibition of LPS response. Antagonistic effects of dihydrochalcones against LPS was mainly observed in glycerophospholipid and sphingolipid metabolism besides promoting amino acid biosynthesis. Moreover, PT showed greater metabolic activity than PZ. Overall, the findings of this study yielded knowledge about the mechanisms of action PZ and PT at metabolic level in modulating inflammatory response in human cells.

Wild boar (Sus scrofa) as bioindicator for environmental exposure to organic pollutants.

Organic pollutants (OPs) are widely distributed around the globe, their presence has become an issue of great concern in the last years due to their potential health effects. Wildlife biomonitoring of OPs has been nowadays a common approach to assess chemical exposure in wildlife and humans. In a sample of 60 wild boars (Sus scrofa) from NW Spain, we evaluated the suitability of using liver and hair samples for the assessment of exposure and bioaccumulation of polychlorinated biphenyls (PCBs) including dioxin and non-dioxin like PCBs (DLPCBs and NDLPCBs), organochlorine and organophosphate pesticides (OCPs and OPPs, respectively), polybromodiphenyl ethers (PBDEs), pyrethroids (PYRs) and polycyclic aromatic hydrocarbons (PAHs). Selective pressurized liquid extraction (SPLE) and liquid-liquid extraction (LLE) methodologies were used to determine the target OPs in liver and hair samples. Clean-up of extracts was performed by solid-phase extraction (SPE) using EZ-POP cartridges and detection by gas chromatography coupled to tandem mass spectrometry (GC-QqQ-MS/MS). We found a distribution pattern of ΣPAHs > ΣOCPs > ΣPYRs > ΣNDLPCBs > ΣOPPs > ΣDLPCBs > ΣPBDEs in liver and of ΣPAHs > ΣOCPs > ΣNDLPCBs > ΣPYRs > ΣOPPs > ΣDLPCBs > ΣPBDEs in hair. Significant correlations (p < 0.050) between the socio-demographic characteristics (gender, age and place) and OP concentrations of OCPs, PBDEs, PYRs, OPPs and PAHs were detected. Moderate correlation was found between HCB, PCB28, PCB157 and chlorpyrifos contents in both hair and liver samples. On the basis of these results, our data shows the correlation and complementary information given by both biological samples.

Rapid liquid chromatographic method for the control of doxycycline and tiamulin residues and their metabolites in vivo assays with pigs: Treatment and depletion.

Results gained from in vivo metabolism could be more authentic to reflect the biotransformation of drugs than in vitro metabolism. This paper discusses the development and characterization of an analytical method for controlling tiamulin (TIA) and doxycycline (DOX) residues in pig plasma and urine, as well as their in vivo biotransformation during treatment and suppression steps at pig farms. For such a purpose, urine samples were taken daily (24-hr intervals) during treatment and until day 8 after last dose. Plasma samples were collected on the last treatment day and up to days 4 and 7 after the last dose. Only traces of TIA and their metabolites have been still detected 8 days after the last day of therapy, in agreement with the period of suppression of both antibiotics set by the manufacturers in 7 days. It was estimated that TIA and DOX half-lives were of two days and one day, respectively. TIA and DOX metabolites have been identified for the first time in plasma and urine pig samples during treatment and suppression steps.

A Metabolomics Approach Reveals Immunomodulatory Effects of Proteinaceous Molecules Derived From Gut Bacteria Over Human Peripheral Blood Mononuclear Cells.

There are strong evidences that probiotics influence the immune status of the host, in a strain-specific manner, acting in the gastrointestinal tract. On the hypothesis that certain extracellular proteins and peptides from gut bacteria may mediate part of this immunomodulation and assuming they are able to diffuse through the mucus layer and interact with immune cells we have developed this work. Our study attempts to understand the immunomodulatory mechanisms of (i) Pext, the extracellular protein fraction of Lactobacillus acidophilus DSM20079T, (ii) HM14, a peptide encrypted in an extracellular glycoside hydrolase from Bifidobacterium longum NCIMB 8809 and (iii) Escherichia coli O111:B4 lipopolysaccharide (LPS), a well-known pro-inflammatory molecule, over human peripheral blood mononuclear cells (PBMCs). An untargeted LC-ESI-QTOF-MS metabolomics approach was applied to reveal intracellular changes in treated-PBMCs isolated from healthy donors. Differences in NADH arrest, NAD+ concentration reduction, as well as increases in palmitic acid and methanephrin were observed in HM14 and Pext treated-cells compared to those stimulated with LPS. This would support an anti-inflammatory molecular mechanism of action of such proteinaceous molecules. Moreover, this methodology has confirms the importance of metabolomics approaches to better understanding immune cell responses to gut bacterial-derived molecules.

Screening of organic pollutants in pet hair samples and the significance of environmental factors.

Organic pollutants (OPs) represent a wide range of chemicals that are potentially harmful for human and wildlife health. Many of these pollutants have been identified as endocrine disruptors that can alter hormonal balance producing adverse biological effects such as neurotoxicity, reproductive disorders, carcinogenicity and hepatotoxicity. For years, hair has been selected as a non-invasive source to assess levels of animal contamination. In the present study, a multiclass screening method for determining about 60 organic pollutants in pet hair was designed and validated for qualitative and quantitative purposes. Concentrations from different classes of organochlorine, and organophosphate pesticides (OCPs, and OPPs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (NDL-PCBs and DL-PCBs), polybrominated diphenyl ethers (PBDEs) and organophosphate esters (OPEs) were identified in the selected pet hair samples from Ourense (NW, Spain). We detected most of these pollutants in the selected hair pets. The mean concentrations found ranged from 89 to 6556ng/g for OPEs, from 8.6 to 1031ng/g for PAHs, from 8.6 to 256ng/g for PBDEs, from 29 to 184ng/g for OPPs, from 0.29 to 139 for OCPs, from 0.30 to 59ng/g for NDL-PCBs and from 1.2 to 14ng/g for DL-PCBs. To our knowledge, this is the first study to document the presence of OPs in pets from North-West Spain and it could provide baseline information for future monitoring of OPs in the area.