Serum Metabolomics and Proteomics to Study the Antihypertensive Effect of Protein Extracts from Tenebrio molitor

Hypertension is the leading risk factor for premature death worldwide and significantly contributes to the development of all major cardiovascular disease events. The management of high blood pressure includes lifestyle changes and treatment with antihypertensive drugs. Recently, it was demonstrated that a diet supplemented with Tenebrio molitor (TM) extracts is useful in the management of numerous pathologies, including hypertension. This study is aimed at unveiling the underlying mechanism and the molecular targets of intervention of TM dietary supplementation in hypertension treatment by means of proteomics and metabolomics techniques based on liquid chromatography coupled with high-resolution mass spectrometry. We demonstrate that serum proteome and metabolome of spontaneously hypertensive rats are severely altered with respect to their normotensive counterparts. Additionally, our results reveal that a diet enriched with TM extracts restores the expression of 15 metabolites and 17 proteins mainly involved in biological pathways associated with blood pressure maintenance, such as the renin-angiotensin and kallikrein-kinin systems, serin protease inhibitors, reactive oxygen scavenging, and lipid peroxidation. This study provides novel insights into the molecular pathways that may underlie the beneficial effects of TM, thus corroborating that TM could be proposed as a helpful functional food supplement in the treatment of hypertension

Doxycycline and sulfadimethoxine transfer from cross-contaminated feed to chicken tissues

International audienceDuring feed preparation at feed mills or during feed mixing in bins at farms, the accidental feed contamination at trace levels by veterinary drug residues, commonly known as carry over, can accidentally but frequently occur. To evaluate the antimicrobial residual concentrations in poultry edible tissues due to contaminated feed, sulfadimethoxine and doxycycline were administered for ten days to chickens by poultry feed incurred at the contamination level frequently recovered during national feed monitoring plans (1-5 mg/kg). Sulfadimethoxine and doxycycline residual concentrations detected in muscle

SIMULTANEOUS DETERMINATION OF LINCOMYCIN AND FIVE MACROLIDE ANTIBIOTIC RESIDUES IN HONEY BY LIQUID CHROMATOGRAPHY COUPLED TO ELECTROSPRAY IONISATION MASS SPECTROMETRY

International audienceA sensitive and specific method based on liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), for the simultaneous determination of lincomycin and five macrolide antibiotics in honey, was developed and validated. The analytes were extracted with Tris buffer 0.1 M pH 10.5 and cleaned-up by a single solid phase extraction step on OASIS HLB column. The chromatographic separation of analytes was performed on a Synergi Hydro-RP reversed – phase column using a gradient program of aqueous 0.01 M ammonium acetate pH 3.5 and acetonitrile as the mobile phase, at a flow rate 0.25 ml min –1. Detection of analytes was achieved by positive ionisation electrospray in Multiple Reaction Monitoring (MRM) mode. Two characteristic transitions were monitored for each substance. The following analytical parameters were validated according to the guidelines laid down by Commission Decision 2002/657/EC: linearity, specificity, decision limit (Cca), detection capability (Ccb), repeatability, within-laboratory reproducibility, recovery and ruggedness

LC-HRMS-Based Non-Targeted Metabolomics for the Assessment of Honey Adulteration with Sugar Syrups: A Preliminary Study

Honey is a natural product that is in great demand and has a relatively high price, thus making it one of the most common targets of economically motivated adulteration. Its adulteration can be obtained by adding cheaper honey or sugar syrups or by overfeeding honeybees with sugar syrups. Adulteration techniques are constantly evolving and advanced techniques and instruments are required for its detection. We used non-targeted metabolomics to underscore potential markers of honey adulteration with sugar syrups. The metabolomic profiles of unadulterated honeys and sugar beet, corn and wheat syrups were obtained using hydrophilic interaction liquid chromatography high-resolution mass spectrometry (LC-HRMS). The potential markers have been selected after data processing. Fortified honey (5%, 10% and 20%), honey obtained from overfeeding, and 58 commercial honeys were analyzed. One potential marker appeared with a specific signal for syrups and not for honey. This targeted analysis showed a linear trend in fortified honeys with a calculated limit of quantification around 5% of fortification

Targeted proteomics for the indirect detection of dexamethasone treatment in bovines

The illegal use of pharmacologically active compounds for growth promotion in food-producing species poses risks for consumer health and animal welfare. Surveillance relies on the quantification of drug residues in animal fluids or tissues, but the efficacy can be negatively affected due to undetectable residual concentrations in biological matrices. Consequently, techniques focusing on the indirect biological effects of exogenous compound administration have been proposed as more sensitive detection methods. The purpose of the present study is to develop a tandem mass spectrometry analytical method based on low-energy collision-induced dissociation (CID-MS/MS) using multiple reaction monitoring (MRM) for the quantification of 12 potential protein markers of skeletal muscle to detect anabolic treatments with dexamethasone. Protein markers identified in a previous study applying a 2D-DIGE proteomics approach have been quantified using the signature peptide method. A group of proteins were confirmed as reliable markers. Quantitative results enabled a predictive model to be defined based on logistic regression for the detection of treated animals. The developed model was finally cross-validated in an independent animal set. [Figure not available: see fulltext.

Bioaccumulation and in vivo formation of titanium dioxide nanoparticles in edible mussels

The aim of this study was to evaluate the bioaccumulation of titanium dioxide nanoparticles (TiO2NPs) in edible mussels bred in polluted artificial seawater. An in vivo study was conducted by exposing mussels to different concentrations of TiO2NPs (0.25 mg/L and 2.5 mg/L) or ionic titanium (1.6 mg/L) for 4 days. Inductively coupled plasma mass spectrometry (ICP-MS) showed titanium presence in all groups proportional to exposure levels (concentration range: 209–1119 µg/kg). Single particle ICP-MS revealed NPs in both TiO2NP treated mussels (concentration range: 231–1778 µg/kg) and in ionic titanium treated mussels (concentration 1574 µg/kg), suggesting potential nanoparticle formation in vivo. These results were confirmed by transmission electron microscopy with energy dispersive X-ray detection. Nonetheless, mussels eliminated more than 70% of the TiO2NPs after 3 days’ depuration. These results show the potential for consumer exposure to TiO2NPs when contaminated mussels are consumed without a proper depuration process

Serum Metabolomics and Proteomics to Study the Antihypertensive Effect of Protein Extracts from Tenebrio molitor

Hypertension is the leading risk factor for premature death worldwide and significantly contributes to the development of all major cardiovascular disease events. The management of high blood pressure includes lifestyle changes and treatment with antihypertensive drugs. Recently, it was demonstrated that a diet supplemented with Tenebrio molitor (TM) extracts is useful in the management of numerous pathologies, including hypertension. This study is aimed at unveiling the underlying mechanism and the molecular targets of intervention of TM dietary supplementation in hypertension treatment by means of proteomics and metabolomics techniques based on liquid chromatography coupled with high-resolution mass spectrometry. We demonstrate that serum proteome and metabolome of spontaneously hypertensive rats are severely altered with respect to their normotensive counterparts. Additionally, our results reveal that a diet enriched with TM extracts restores the expression of 15 metabolites and 17 proteins mainly involved in biological pathways associated with blood pressure maintenance, such as the renin-angiotensin and kallikrein-kinin systems, serin protease inhibitors, reactive oxygen scavenging, and lipid peroxidation. This study provides novel insights into the molecular pathways that may underlie the beneficial effects of TM, thus corroborating that TM could be proposed as a helpful functional food supplement in the treatment of hypertension

The Prion Protein Regulates Synaptic Transmission by Controlling the Expression of Proteins Key to Synaptic Vesicle Recycling and Exocytosis.

The cellular prion protein (PrP C ), whose misfolded conformers are implicated in prion diseases, localizes to both the presynaptic membrane and postsynaptic density. To explore possible molecular contributions of PrP C to synaptic transmission, we utilized a mass spectrometry approach to quantify the release of glutamate from primary cerebellar granule neurons (CGN) expressing, or deprived of (PrP-KO), PrP C , following a depolarizing stimulus. Under the same conditions, we also tracked recycling of synaptic vesicles (SVs) in the two neuronal populations. We found that in PrP-KO CGN these processes decreased by 40 and 60%, respectively, compared to PrP C -expressing neurons. Unbiased quantitative mass spectrometry was then employed to compare the whole proteome of CGN with the two PrP genotypes. This approach allowed us to assess that, relative to the PrP C -expressing counterpart, the absence of PrP C modified the protein expression profile, including diminution of some components of SV recycling and fusion machinery. Subsequent quantitative RT-PCR closely reproduced proteomic data, indicating that PrP C is committed to ensuring optimal synaptic transmission by regulating genes involved in SV dynamics and neurotransmitter release. These novel molecular and cellular aspects of PrP C add insight into the underlying mechanisms for synaptic dysfunctions occurring in neurodegenerative disorders in which a compromised PrP C is likely to intervene