Glyphosate presence in human sperm: First report and positive correlation with oxidative stress in an infertile French population.

Environmental exposure to endocrine disruptors, such as pesticides, could contribute to a decline of human fertility. Glyphosate (GLY) is the main component of Glyphosate Based Herbicides (GBHs), which are the most commonly herbicides used in the world. Various animal model studies demonstrated its reprotoxicity. In Europe, GLY authorization in agriculture has been extended until 2034. Meanwhile the toxicity of GLY in humans is still in debate. The aims of our study were firstly to analyse the concentration of GLY and its main metabolite, amino-methyl-phosphonic acid (AMPA) by LC/MS-MS in the seminal and blood plasma in an infertile French men population (n=128). We secondly determined Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) using commercial colorimetric kits and some oxidative stress biomarkers including malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) by ELISA assays. We next analysed potential correlations between GLY and oxidative stress biomarkers concentration and sperm parameters (sperm concentration, progressive speed, anormal forms). Here, we detected for the first time GLY in the human seminal plasma in significant proportions and we showed that its concentration was four times higher than those observed in blood plasma. At the opposite, AMPA was undetectable. We also observed a strong positive correlation between plasma blood GLY concentrations and plasma seminal GLY and 8-OHdG concentrations, the latter reflecting DNA impact. In addition, TOS, Oxidative Stress Index (OSI) (TOS/TAS), MDA blood and seminal plasma concentrations were significantly higher in men with glyphosate in blood and seminal plasma, respectively. Taken together, our results suggest a negative impact of GLY on the human reproductive health and possibly on his progeny. A precaution principle should be applied at the time of the actual discussion of GLY and GBHs formulants uses in Europe by the authorities.

Hexahydrocannabinol poisoning reported to French poison centres.

INTRODUCTION: Hexahydrocannabinol is a hexahydro derivative of cannabinol. Poisoning with hexahydrocannabinol was first observed in Europe in May 2022. METHOD: This is a retrospective observational study of cases of self-reported hexahydrocannabinol exposure reported to French poison centres between 1 January 2022 and 31 May 2023. RESULTS: There were 37 cases, including 19 in May 2023. The median age of the patients was 36 (interquartile range 28-43) years, and most were men. Eight patients had a history of substance use disorder. The route of exposure was ingestion in 24, inhalation (smoking or vaping) in 10, inhalation and ingestion in two and sublingual in one. Clinical features were neurological (85 per cent), cardiovascular (61 per cent), gastrointestinal (33 per cent), psychiatric (27 per cent) and ocular (21 per cent). Fifty-nine per cent of the patients were hospitalized. In four patients, the Poisoning Severity Score was 0 (asymptomatic); in 15 patients, the Score was 1 (minor); in 16, the Score was 2 (moderate); and in two cases, the Score was 3 (severe). In 70 per cent of patients, the outcome was known, and all recovered. Testing of biological samples was only undertaken in six cases. Five patients had positive blood or urine tests for hexahydrocannabinol; in two patients, tetrahydrocannabinol and metabolites were also detected. In addition, there was an additional patient in whom Δ8- and Δ9-tetrahydrocannabinol was detected in the substances used. DISCUSSION: Clinical effects reported in this series included neuropsychiatric and somatic effects. Whilst these cases related to self-reported hexahydrocannabinol use, it is likely that tetrahydrocannabinol use also contributed to the effects in a substantial proportion of cases. This study has some limitations, such as the lack of available information due to the retrospective nature of the study. As a result, it probably overestimates the number of moderate and severe cases due to under-reporting of cases of little or no severity. Analysis of the patient's blood and urine was performed only in six patients, so we cannot be certain that the products consumed by the other patients were hexahydrocannabinol. CONCLUSION: The clinical effects attributed to hexahydrocannabinol were neurological, cardiovascular, gastrointestinal, psychiatric and ocular predominantly and were sometimes serious.

Probe Electrospray Ionization Tandem Mass Spectrometry for the Detection and Quantification of Benzodiazepines.

BACKGROUND: Legally prescribed benzodiazepines (BZDs) and designer BZDs are widely misused and must be determined in multiple contexts (eg, overdose, drug-facilitated sexual assaults, or driving under the influence of drugs). This study aimed to develop a method for measuring serum BZD levels using probe electrospray ionization (PESI) mass spectrometry and an isotope dilution approach. METHODS: A tandem mass spectrometer equipped with a probe electrospray ionization source in multiple reaction monitoring mode was used. Isotope dilution was applied for quantification using a deuterated internal standard at a fixed concentration for alprazolam, bromazepam, diazepam, nordiazepam, oxazepam, temazepam, zolpidem, and zopiclone. This method included designer BZDs: clonazolam, deschloroetizolam, diclazepam, etizolam, flualprazolam, flubromazepam, flubromazolam, meclonazepam, nifoxipam, and pyrazolam. Sample preparation was done by mixing 10 µL of serum with 500 µL of an ethanol/ammonium formate 0.01 mol/L buffer. Complete validation was performed, and the method was compared with liquid chromatography coupled with mass spectrometry (LC-MS/MS) and immunoassays (IC) by analyzing 40 real samples. RESULTS: The analysis time for identification and quantification of the 18 molecules was 2.5 minutes. This method was fully validated, and the limits of quantification varied from 5 to 50 mcg/L depending on the molecule. In the 40 real samples, 100% of molecules (n = 89) were detected by both LC-MS/MS and PESI-MS/MS, and regression analysis showed excellent agreement between the 2 methods (r2 = 0.98). On IC, bromazepam and zolpidem were not detected in 2 and 1 cases, respectively. CONCLUSIONS: PESI-MS/MS allows serum BZD detection and measurement. Given the isotope dilution approach, a calibration curve was not required, and its performance was similar to that of LC-MS/MS, and its specificity was higher than that of IC.

Early detection of liver injuries by the Serum enhanced binding test sensitive to albumin post-transcriptional modifications.

Early and sensitive biomarkers of liver dysfunction and drug-induced liver injury (DILI) are still needed, both for patient care and drug development. We developed the Serum Enhanced Binding (SEB) test to reveal post-transcriptional modifications (PTMs) of human serum albumin resulting from hepatocyte dysfunctions and further evaluated its performance in an animal model. The SEB test consists in spiking serum ex-vivo with ligands having specific binding sites related to the most relevant albumin PTMs and measuring their unbound fraction. To explore the hypothesis that albumin PTMs occur early during liver injury and can also be detected by the SEB test, we induced hepatotoxicity in male albino Wistar rats by administering high daily doses of ethanol and CCl4 over several days. Blood was collected for characterization and quantification of albumin isoforms by high-resolution mass spectrometry, for classical biochemical analyses as well as to apply the SEB test. In the exposed rats, the appearance of albumin isoforms paralleled the positivity of the SEB test ligands and histological injuries. These were observed as early as D3 in the Ethanol and CCl4 groups, whereas the classical liver tests (ALT, AST, PAL) significantly increased only at D7. The behavior of several ligands was supported by structural and molecular simulation analysis. The SEB test and albumin isoforms revealed hepatocyte damage early, before the current biochemical biomarkers. The SEB test should be easier to implement in the clinics than albumin isoform profiling.

Absolute Quantification of Human Serum Albumin Isoforms by Internal Calibration Based on a Top-Down LC-MS Approach.

Well-characterized biomarkers using reliable quantitative methods are essential for the management of various pathologies such as diabetes, kidney, and liver diseases. Human serum albumin (HSA) isoforms are gaining interest as biomarkers of advanced liver pathologies. In view of the structural alterations observed for HSA, insights into its isoforms are required to establish them as reliable biomarkers. Therefore, a robust absolute quantification method seems necessary. In this study, we developed and validated a far more advanced top-down liquid chromatography-mass spectrometry (LC-MS) method for the absolute quantification of HSA isoforms, using myoglobin (Mb) as an internal standard for quantification and for mass recalibration. Two different quantification approaches were investigated based on peak integration from the deconvoluted spectrum and extracted ion chromatogram (XIC). The protein mixture human serum albumin/myoglobin eluted in well-shaped separated peaks. Mb allowed a systematic mass recalibration for every sample, resulting in extremely low mass deviations compared to conventional deconvolution-based methods. In total, eight HSA isoforms of interest were quantified. Specific-isoform calibration curves showing good linearity were obtained by using the deconvoluted peaks. Noticeably, the HSA ionization behavior appeared to be isoform-dependent, suggesting that the use of an enriched isoform solution as a calibration standard for absolute quantification studies of HSA isoforms is necessary. Good repeatability, reproducibility, and accuracy were observed, with better sensitivity for samples with low albumin concentrations compared to routine biochemical assays. With a relatively simple workflow, the application of this method for absolute quantification shows great potential, especially for HSA isoform studies in a clinical context, where a high-throughput method and sensitivity are needed.

Chlordecone determination in serum by LC-MS/MS and the importance of low limit of detection.

Chlordecone is an organochlorine insecticide that has been used intensively from 1973 to 1993 in the French West Indies banana fields to control root borers. This use has resulted in persistent pollution of soils and waters, and people have been and are still exposed mainly through food. Epidemiological studies showed that this exposure is associated with health disorders, including prostate cancer, prematurity, cognitive or motor development. The measurement of chlordecone in serum is considered as the best surrogate, though no clear and definitive cut-off value has been established. This renders necessary the development of analytical methods with the lowest limit of detection as possible. While most published methods have utilized GC-MS or GC-MS/MS, in the present study we report an LC-MS/MS method based on a simple QuEChERS salts extraction. The whole procedure was validated according to ISO 15189 requirements and reached LOD and LOQ values of 0.007 and 0.02 µg/L, respectively. It was applied to more 10 000 serum samples of French Indies inhabitants. More than a half had a concentration below 0.1 µg/L and more than one third of them exhibiting a concentration below 0.05 µg/L. The capability of this LC-MS/MS method to detect very low concentrations highlights its utility in exploring the health impact of chlordecone.

Comparison of homocitrulline and carbamylated albumin as biomarkers of carbamylation reactions in hemodialyzed patients.

To describe the association between levels of homocitrulline (HCit) and the degree of albumin carbamylation in a cohort of hemodialyzed patients. Plasma total and protein-bound HCit concentrations in samples from hemodialyzed patients included in NICOREN trial were determined by LC-MS/MS at baseline and after 24 weeks of treatment with either sevelamer or nicotinamide. HCit concentrations at all timepoints and in both groups were positively and significantly correlated with the degree of albumin carbamylation. Plasma concentrations of total HCit, protein-bound HCit and carbamylated albumin did not decrease after 24 weeks of treatment with either sevelamer or nicotinamide. The present results demonstrate that plasma total and protein-bound HCit concentrations were closely associated with albumin carbamylation in hemodialyzed patients. Therefore, total and protein-bound HCit concentrations might be valuable biomarkers of the overall intensity of protein carbamylation in this context. Given the less complex and time-consuming analytical methods required, these markers should be favored in future clinical studies of carbamylation reaction.

Semi-synthetic human albumin isoforms: Production, structure, binding capacities and influence on a routine laboratory test.

Human Serum Albumin (HSA) undergoes Post-Translational-Modifications (PTMs) leading to isoforms affecting its oncotic and non-oncotic properties. HSA is comprised of several isoforms whose abundance may vary with pathologies such as diabetes, kidney and liver diseases. Studying their impact separately may help to understand their sources and potential pathogenicity and further their evaluation as biomarkers. The present study examined semi-synthetic HSA isoforms to investigate independently their structure by means of advanced mass spectrometry techniques (LC-TOF-MS and ICP-MS), influence on the HSA binding/antioxidant activities using a binding capacity test, and potential impact on albumin quantification by a routine immunoturbidimetric assay. Applying different chemical reactions to a commercial HSA solution, we obtained different solutions enriched up to 53 % of native HSA, 78 % of acetylated HSA, 71 % of cysteinylated HSA, 94 % of oxidized HSA, 58 % of nitrosylated HSA and 96 % of glycated HSA, respectively. Moreover, the semi-synthetic isoforms showed differently altered binding capacities for a panel of ligands (Cu, Cd, Au, Ds and L-T4). Furthermore, immunoturbidimetry was found to be insensitive to the presence and abundance of the different isoforms. The fully characterized semi synthetic HSA isoforms obtained should be useful to further investigate their pathogenicity and potential roles as biomarkers.

Posttranslational-modifications of human-serum-albumin analysis by a top-down approach validated by a comprehensive bottom-up analysis.

The posttranslational modifications (PTM) of human serum albumin (HSA) can result in the development of isoforms that have been identified as potential biomarkers for advanced hepatic diseases. However, previous approaches using top-down (TD) analysis to identify isoforms based on molecular weight may have resulted in misidentifications. The nature of the identified isoforms has never been confirmed in previous works. Here, we aimed to critically evaluate TD for the characterization and determination of HSA isoforms in patients and make an inventory of HSA-PTM. Serum samples from control subjects and patients with liver dysfunctions were analyzed using both top-down (TD) and bottom-up (BU) approaches. TD analysis involved using a LC-TOF-MS system to obtain a multicharged spectrum of HSA, which was deconvoluted to identify isoforms. Spectra were then used for relative quantitation analysis of albumin isoform abundances based on trapezoidal integration. For BU analysis, serums were reduced +/- alkylated, digested with trypsin and analyzed in the Q-TOF, data-dependent acquisition (DDA) mode to generate a SWATH-MS high-resolution mass spectral library of all HSA peptides. Tryptic digests of another set of serum samples were then analyzed using data-independent acquisition (DIA) mode to confirm the presence of HSA isoforms and their modification sites. TD detected 15 isoforms corresponding to various modifications, including glycation, cysteinylation, nitrosylation, and oxidation (di- and tri-). In BU, the spectral library containing 127 peptides allowed for the characterization of the important isoforms with their modified sites, including some modifications that were only characterized in BU (carbamylation, deamidation, and amino-acid substitution). The method used for determining isoforms offered acceptable reproducibility (intra-/inter-assay CVs < 15%) for all isoforms present at relative abundances higher than 2%. Overall, the study found that several isoforms could be missed or misidentified by TD. However, all HSA isoforms identified by TD and reported to be relevant in liver dysfunctions were confirmed by BU. This critical evaluation of TD approach helped design an adequate and reliable method for the characterization of HSA isoforms in patients and offers the possibility to estimate isoform abundances within 3 min. These findings have significant implications for the diagnosis and treatment of liver dysfunctions.

Optimization of ICP-MS internal standardization for 26 elements by factorial design experiment.

INTRODUCTION: Internal standardization is a common tool used in inductively coupled plasma - mass spectrometry (ICP-MS) analysis in order to reduce matrix effects, and thus improve the reliability and the robustness of results. However, its efficacy relies on the choice of a proper internal standard (IS) which, ideally, undergoes the same signal variations as the analyte. Thus far, IS selection has mainly relied on the proximity of atomic mass between the analyte and the internal standard. However, while it may be a satisfactory rule of thumb, more recent works suggest that this criterium might not be suitable in several conditions, among which the presence of high amounts of carbon atoms in the sample. This may thus be of particular interest in the case of trace elements determination in biological samples. MATERIALS AND METHODS: In this study, we propose an empirical and global approach to IS selection in ICP-MS through the use of a factorial design of experiments (DoE), with a focus on biological matrices of interest in clinical analysis: human blood and urine. The suitability of 13 potential IS was evaluated for 26 clinically-relevant analytes, including a polyatomic ion obtained through reaction with oxygen, across 324 experimental conditions. RESULTS AND DISCUSSION: The results underline several exceptions to the rule of IS selection based on mass proximity, notably when considering heavy or polyatomic analytes. As a consequence, measurements of said analytes in several extreme experimental conditions using IS selected by mass proximity could yield vastly erroneous results (up to 30 times the theoretical concentrations). By contrast, the use of empirically selected IS yielded much more acceptable results.

New perspectives for the therapeutic drug monitoring of tacrolimus: Quantification in volumetric DBS based on an automated extraction and LC-MS/MS analysis.

Volumetric microsampling devices have been developed for home-based capillary blood sampling and are now increasingly proposed for the therapeutic drug monitoring (TDM) of immunosuppressive drugs. Our objective was to validate a LC-MS/MS method for tacrolimus quantification based on both a manual and an automated extraction of dried blood spots (DBS) collected with a volumetric microsampling device. DBS collection was performed by placing a drop of whole blood (WB) pre-spiked with tacrolimus onto a sealing film and placing the hemaPEN® device (Trajan Scientific and Medical, Melbourne, Australia) into the drop according to the device specifications. Tacrolimus was quantified using a fully automatic preparation module connected to a LCMS system (CLAM-3020® and LCMS-8060®, Shimadzu, Marne-la-Vallée, France). The method was validated analytically and clinically in accordance with the EMA and IATDMCT guidelines. The method was linear from 1 to 100 µg/L. Within- and between-run accuracy and precision fulfilled the validation criteria (biases and imprecision <15% or 20% for the lower limit of quantification). No hematocrit effect, matrix effect or carry-over was observed. No selectivity issue was identified and dilution integrity was confirmed. Tacrolimus in DBS was stable for 14 days at room temperature and +4°C, and for 72h at +60°C. There was a good correlation between tacrolimus concentrations measured in WB and in DBS of 20 kidney and liver transplant recipients (r=0.93 and 0.87, for manual and automated extraction respectively). A method for tacrolimus measurement in DBS collected with volumetric micro-sampling device, based on a fully automated process from pre-treatment to LC-MS/MS analysis was developed and validated according to analytical and clinical criteria. This performing sampling and analytical procedure opens the perspective of an easier, faster and more efficient TDM of tacrolimus for patients, clinicians and laboratories.

Designer Benzodiazepines: Effects, Toxicity, and Interactions.

PURPOSE: Although designer benzodiazepines (DBZDs) constitute a minor part of new psychoactive substances, they deserve the greatest attention because of their popularity among drug users and increasing number and availability. This review covers the effects of different DBZDs, available pharmacological evaluation tools, and their reported toxicity and potential pharmacodynamic and pharmacokinetic interactions with other drugs commonly co-abused with DBZDs. METHODS: For this narrative review, a nonsystematic search was performed on PubMed, EMBASE, Google Scholar, and PubMed Central databases between June and July 2021. RESULTS: The current consensus hypothesis suggests that DBZDs mediate their effects through interactions with the GABA A receptor, producing similar effects to benzodiazepines used in therapy, including sedation, hypnosis, anxiolysis, muscle relaxation, euphoria, amnesia, and addiction. Owing to the complexity of their action mechanism and the numerous GABA A subtype receptors, the pharmacodynamic metrics of DBZDs are very difficult to establish. The pharmacological effects of DBZD are related to their structure, influencing their binding to GABA A receptor subunits. Quantitative structure-activity relationship studies successfully predicted the biological activity and relative potency of DBZD but could not predict the main pharmacological effect of a given DBZD. Exploring the effects by netnographic studies is one of the available alternatives, despite its limitations. DBZDs are usually identified in the context of polysubstance use. Pharmacodynamic interactions between DBZDS and other CNS depressants, such as opioids, have been extensively reported. However, pharmacokinetic interactions between DBZDs and opioids are considered less important, and contradictory conclusions about their clinical significance have been reported. CONCLUSIONS: Understanding the mechanism of action and other pharmacological metrics is highly important in the clinical management of DBZDs.

Metabolic interactions of benzodiazepines with oxycodone ex vivo and toxicity depending on usage patterns in an animal model.

BACKGROUND AND PURPOSE: Opioids and benzodiazepines are frequently combined in medical as well as in non-medical contexts. At high doses, such combinations often result in serious health complications attributed to pharmacodynamics interactions. Here, we investigate the contribution of the metabolic interactions between oxycodone, diazepam and diclazepam (a designer benzodiazepine) in abuse/overdose conditions through ex vivo, in vivo and in silico approaches. EXPERIMENTAL APPROACH: A preparation of pooled human liver microsomes was used to study oxycodone metabolism in the presence or absence of diazepam or diclazepam. In mice, diazepam or diclazepam was concomitantly administered with oxycodone to mimic acute intoxication. Diclazepam was introduced on Day 10 in mice continuously infused with oxycodone for 15 days to mimic chronic intoxication. In silico modelling was used to study the molecular interactions of the three drugs with CYP3A4 and 2D6. KEY RESULTS: In mice, in acute conditions, both diazepam and diclazepam inhibited the metabolism of oxycodone. In chronic conditions and at pharmacologically equivalent doses, diclazepam drastically enhanced the production of oxymorphone. In silico, the affinity of benzodiazepines was higher than oxycodone for CYP3A4, inhibiting oxycodone metabolism through CYP3A4. Oxycodone metabolism is likely to be diverted towards CYP2D6. CONCLUSION AND IMPLICATIONS: Acute doses of diazepam or diclazepam result in the accumulation of oxycodone, whereas chronic administration induces the accumulation of oxymorphone, the toxic metabolite. This suggests that overdoses of opioids in the presence of benzodiazepines are partly due to metabolic interactions, which in turn explain the patterns of toxicity dependent on usage. LINKED ARTICLES: This article is part of a themed issue on Advances in Opioid Pharmacology at the Time of the Opioid Epidemic. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.7/issuetoc.

Ultrafast Measurement of Metformin in the Clinical Setting Using Probe Electrospray Ionization Mass Spectrometry.

Metformin (MtF) is a treatment used for type 2 diabetes. Lactic acidosis (LA) is a frequent complication that can be either induced by or associated with elevated MtF plasma concentrations. When coupled with a mass spectrometry (MS) system, the probe electrospray ionization (PESI) method allows direct and rapid analysis of different types of matrices without pretreatment. In this study, we developed a PESI-MS method for the determination of MtF in plasma. We used a tandem mass spectrometer equipped with a PESI source in the reaction monitoring mode for the quantitation of MtF. MtF-d6 was chosen as the internal standard (IS), following an isotope dilution (ID) approach. The method was fully validated with six concentration levels (0.5-50 mg/L). The matrix effect was evaluated for each level, and the specificity was tested with a mix of potential co-medications. Using patient samples, the performance was compared with two classical LC-MS-MS and LC-diode array detector (DAD) methods used in external labs. Sample preparation consisted in mixing 10 µL plasma in 1,000 µL ethanol/ammonium formate buffer including MtF-d6 at a fixed concentration of 5 mg/L. The total run time was 0.31 min. ID gave satisfactory results of accuracy and precision (min-max: -12.1 to 15.8% and 1.0-17.1%, respectively). The matrix effect was fully corrected by the internal standard (bias < 1%). The specificity study also reported satisfactory results. Finally, in a representative group of 29 patients (55% with a concentration <5 mg/L, 38% with a concentration >5 mg/L and 7% not detected), we observed almost identical results when comparing LC-DAD and LC-MS-MS to PESI-MS (r2 > 0.99). We propose a specific, sensitive, accurate and ultrafast solution for the measurement of MtF in patient plasma, with no sample preparation or calibration curve building. This could be helpful in a core lab when rapid diagnosis of LA is needed.

Urine toxicological screening method using an immunoenzymatic assay based on a biochip array technology: comparative study with immunochromatography and Liquid chromatography­mass spectrometry / Criblage toxicologique urinaire par une méthode automatisée immunoenzymatique multiparamétrique basée sur une technique de biopuce : comparaison à de l'immunochromatographie et de la chromatographie liquide couplée à de la spectrométrie de masse

Multiparametric toxicology research is mainly based on immunochromatography [IC] and chromatography methods. A new automated method using an immunoenzymatic (IE) assay based on a biochip array technology combines short turning around time and analytical performances close to chromatography in terms of positivity cut-off. The aim of our study was to compare IE versus IC and chromatography methods using urines samples from clinical cases. Seventy-two samples were analyzed by IC (amphetamines, opiates, benzodiazepines, THC, methadone, cocaine), IE and chromatography (previous classes plus opioids and cathinone). Immunochromatography results were read by at least 7 operators to assess reading subjectivity. Immunoenzymatic, IC, and chromatrography results were compared with each other. Chromatographic quantification was analyzed to understand discrepancies. Significant discrepancies (29-64%) were observed between IC and IE for most of the drug families investigated except for benzodiazepines, methadone and opiates. These discrepancies were not identified between IE and chromatography, except for some substances (28% to 67% discrepancies for buprenorphine, tramadol and oxycodone, 100% for cathinone). In contrast to IC, the performance of IE approached those of chromatography, except for some substances for which cross-reactions must be investigated. Reading discrepancies were frequent with IC (33% of samples) and made robust result output challenging. In conclusion, the Multistat® is an interesting method for first-line toxicological screening for laboratories without chromatography method.

La recherche des toxiques multiparamétrique repose principalement sur des méthodes immunochromatographie [IC] et de chromatographie (CL). Une nouvelle méthode automatisée immunoenzymatique (IE) (Multistat®) en biopuce, combine un rendu de résultats rapide et des performances analytiques, en termes de seuils de positivité, proche de la CL. L'objectif de notre étude a été de comparer l'IE à des méthodes d'IC et de CL sur des échantillons hospitaliers. Soixante-douze échantillons ont été analysés par IC (amphétamines, opiacés, benzodiazépines, THC, méthadone, cocaïne), IE et CL (classes précédentes plus opioïdes et dérivés de la cathinone). Les résultats d'IC étaient lus par au moins 7 personnes pour évaluer la subjectivité des lectures. Les résultats d'IE, d'IC et de CL étaient comparés entre eux. La quantification en CL était exploitée pour expliquer les discordances. Une forte proportion de discordances (de 29 à 64 %) était observée entre IC et IE sur la plupart des toxiques explorées sauf pour les benzodiazépines, la méthadone et les opiacés. Ces discordances n'étaient pas retrouvées entre IE et CL, hormis pour certaines substances (28 % à 67 % de divergences pour buprénorphine, tramadol et oxycodone, 100 % pour les dérivés de la cathinone). À l'inverse de l'IC, les performances de l'IE se rapprochaient de celles de la CL, sauf pour certaines substances pour lesquelles des réactions croisées doivent être recherchées. Les discordances de lecture étaient fréquentes en IC et rendent difficile un rendu de résultat robuste. En conclusion, le Multistat® est une méthode intéressante pour un criblage en première intention pour les laboratoires sans CL.

Problematic rise of vitamin B6 supplementation overuse and potential risk to bariatric surgery patients.

OBJECTIVES: Due to the increased prevalence of obesity in the world, bariatric surgeries are on the rise and necessitate life-long surveillance for deficiencies; hence the recommended vitamin supplementation in these patients. However, inadequate multivitamin supplementation may induce vitamin B6 overload. METHODS: We reviewed all vitamin B6 dosages at the university hospitals of Poitiers, Tours, Bordeaux, and Limoges for the past 5 to 8 years. Analyses were performed by high-performance liquid chromatography, coupled with a fluorescence detector on whole blood samples. RESULTS: During the study period, there was an increase in the number of vitamin B6 dosages. Deficiencies were detected early in Poitiers and Limoges, but were negligible by 2020. However, during the same time period, the number of overdoses increased, reaching close to 40% of dosages at all centers. CONCLUSIONS: Pyridoxin overload is not possible through food-derived pyridoxin; hence, combined with the fact that most vitamin supplements contain vitamin B6, inadequate vitamin supplementation is likely the cause of the observed increase in overdoses. High doses of vitamin B6 can induce polyneuropathy, particularly targeting motor neurons; thus, the increase of overdoses is worrying. In light of the possible risks and the ease with which these could be averted (better formulation of supplements), the precaution principle requires a definition of clear guidelines for vitamin supplementation, especially in patients undergoing bariatric surgery.

Impacts of dietary exposure to pesticides on faecal microbiome metabolism in adult twins.

BACKGROUND: Dietary habits have a profound influence on the metabolic activity of gut microorganisms and their influence on health. Concerns have been raised as to whether the consumption of foodstuffs contaminated with pesticides can contribute to the development of chronic disease by affecting the gut microbiome. We performed the first pesticide biomonitoring survey of the British population, and subsequently used the results to perform the first pesticide association study on gut microbiome composition and function from the TwinsUK registry. METHODS: Dietary exposure of 186 common insecticide, herbicide, or fungicide residues and the faecal microbiome in 65 twin pairs in the UK was investigated. We evaluated if dietary habits, geographic location, or the rural/urban environment, are associated with the excretion of pesticide residues. The composition and metabolic activity of faecal microbiota was evaluated using shotgun metagenomics and metabolomics respectively. We performed a targeted urine metabolomics analysis in order to evaluate whether pesticide urinary excretion was also associated with physiological changes. RESULTS: Pyrethroid and/or organophosphorus insecticide residues were found in all urine samples, while the herbicide glyphosate was found in 53% of individuals. Food frequency questionnaires showed that residues from organophosphates were higher with increased consumption of fruit and vegetables. A total of 34 associations between pesticide residue concentrations and faecal metabolite concentrations were detected. Glyphosate excretion was positively associated with an overall increased bacterial species richness, as well as to fatty acid metabolites and phosphate levels. The insecticide metabolite Br2CA, reflecting deltamethrin exposure, was positively associated with the phytoestrogens enterodiol and enterolactone, and negatively associated with some N-methyl amino acids. Urine metabolomics performed on a subset of samples did not reveal associations with the excretion of pesticide residues. CONCLUSIONS: The consumption of conventionally grown fruit and vegetables leads to higher ingestion of pesticides with unknown long-term health consequences. Our results highlight the need for future dietary intervention studies to understand effects of pesticide exposure on the gut microbiome and possible health consequences.

Chronic Dietary Exposure of Roosters to a Glyphosate-Based Herbicide Increases Seminal Plasma Glyphosate and AMPA Concentrations, Alters Sperm Parameters, and Induces Metabolic Disorders in the Progeny.

The effects of chronic dietary Roundup (RU) exposure on rooster sperm parameters, fertility, and offspring are unknown. We investigated the effects of chronic RU dietary exposure (46.8 mg kg-1 day-1 glyphosate) for 5 weeks in 32-week-old roosters (n = 5 RU-exposed and n = 5 control (CT)). Although the concentrations of glyphosate and its main metabolite AMPA (aminomethylphosphonic acid) increased in blood plasma and seminal fluid during exposure, no significant differences in testis weight and sperm concentrations were observed between RU and CT roosters. However, sperm motility was significantly reduced, associated with decreased calcium and ATP concentrations in RU spermatozoa. Plasma testosterone and oestradiol concentrations increased in RU roosters. These negative effects ceased 14 days after RU removal from the diet. Epigenetic analysis showed a global DNA hypomethylation in RU roosters. After artificial insemination of hens (n = 40) with sperm from CT or RU roosters, eggs were collected and artificially incubated. Embryo viability did not differ, but chicks from RU roosters (n = 118) had a higher food consumption, body weight and subcutaneous adipose tissue content. Chronic dietary RU exposure in roosters reduces sperm motility and increases plasma testosterone levels, growth performance, and fattening in offspring.