A rapid LC-MS/MS method for the simultaneous quantification of ivacaftor, lumacaftor, elexacaftor, tezacaftor, hexyl-methyl ivacaftor and ivacaftor carboxylate in human plasma.

Cystic fibrosis is one of the most common genetic diseases among caucasian population. This disease is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene encoding for the CFTR protein. Lumacaftor, elexacaftor, tezacaftor, and ivacaftor were currently used as the treatment to Cystic fibrosis. In this study, we describe a new method for the simultaneous quantification of four molecules: lumacaftor, elexacaftor, tezacaftor, and ivacaftor, alongside two metabolites of ivacaftor, specifically hexyl-methyl ivacaftor and ivacaftor carboxylate by liquid chromatography-tandem mass spectrometry. This method holds significant utility for therapeutic drug monitoring and the optimization of treatments related to CFTR modulators. Molecules were extracted from 100 µL of plasma by a simple method of protein precipitation using acetonitrile. Following extraction, chromatographic separation was carried out by reverse chromatography on a C18 analytical column, using a gradient elution of water (0.05 % formic acid, V/V) and acetonitrile (0.05 % formic acid, V/V). The run time was 7 minutes at a flow rate of 0.5 mL/min. After separation, molecules were detected by electrospray ionization on a Xevo TQD triple-quadrupole-mass-spectrometer (Waters®, Milford, USA). The calibration range were: 0.053-20.000 mg/L for elexacaftor, tezacaftor and lumacaftor, 0.075-14.000 mg/L for ivacaftor, and 0.024-6.500 mg/L for hexyl-methyl ivacaftor and ivacaftor carboxylate. The proposed method underwent throughout validation demonstrating satisfactory precision (inter- and intra-day coefficients of variation less than 14.3 %) and a good accuracy (inter- and intra-day bias ranging between -13.7 % and 14.7 %) for all the analytes. The presented method for the simultaneous quantification of CFTR modulators and their metabolites in human plasma has undergone rigorous validation process yielding good results including strong precision and accuracy for all analytes. This method has been effectively used in routine analytical analysis and clinical investigations within our laboratory.

Lumacaftor/Ivacaftor Population Pharmacokinetics in Pediatric Patients with Cystic Fibrosis: A First Step Toward Personalized Therapy.

BACKGROUND: A major breakthrough in cystic fibrosis (CF) therapy was achievedAQ1 with CFTR modulators. The lumacaftor/ivacaftor combination is indicated for the treatment of CF in pediatric patients above 6 years old. Pharmacokinetic (PK) studies of lumacaftor/ivacaftor in these vulnerable pediatric populations are AQ2crucial to optimize treatment protocols. OBJECTIVES AND METHODS: The objectives of this study were to describe the population PK (PPK) of lumacaftor and ivacaftor in children with CF, and to identify factors associated with interindividual variability. The association between drug exposure and clinical response was also investigated. RESULTS: A total of 75 children were included in this PPK study, with 191 concentrations available for each compound and known metabolites (lumacaftor, ivacaftor, ivacaftor-M1, and ivacaftor-M6). PPK analysis was performed using Monolix software. A large interindividual variability was observed. The main sources of interpatient variability identified were patient bodyweight and hepatic function (aspartate aminotransferase). Forced expiratory volume in the first second (FEV1) was statistically associated with the level of exposure to ivacaftor after 48 weeks of treatment. CONCLUSIONS: This study is the first analysis of lumacaftor/ivacaftor PPK in children with CF. These data suggest that dose adjustment is required after identifying variability factors to optimize efficacy. The use of therapeutic drug monitoring as a basis for dose adjustment in children with CF may be useful.

Impact of perinatal environmental health education intervention on exposure to endocrine disruptors during pregnancy-PREVED study: study protocol for a randomized controlled trial.

BACKGROUND: The suspected or actual effects on health of endocrine-disrupting chemicals (EDC) and their ubiquitous presence in everyday life justify the implementation of health promotion interventions. These interventions should ideally be applied during critical windows like pregnancy. Perinatal environmental health education interventions may help to reduce EDC exposure during pregnancy. METHODS/DESIGN: PREVED (Pregnancy, PreVention, Endocrine Disruptors) is an open-label randomized controlled trial assessing the impact of environmental health education intervention on EDC exposure during pregnancy. Inclusion, consent, and randomization take place during the first trimester. The participants are randomly allocated into three groups: (i) control group (information leaflet on EDCs), (ii) intervention group in neutral location (information leaflet and workshops in a meeting room), and (iii) intervention group in contextualized location (information leaflet and workshops in a real apartment). Workshops are organized between the second and third trimesters of pregnancy. Main outcome is the percentage of participants who reported consuming manufactured/industrial food. Secondary outcomes are as follows: (i) psycho-social dimensions, (ii) EDC concentrations in urine, (iii) EDC concentration in colostrum, and (iv) percentage of participants who reported consuming paraben-free personal care products. DISCUSSION: PREVED is a ground-breaking intervention research project dedicated to perinatal environmental health education that aims to identify pollutant sources in daily life and to offer accessible and realistic alternative solutions, by promoting the sharing of know-how and experience in a positive and non-alarmist approach. TRIAL REGISTRATION: ClinicalTrials.gov : NCT03233984 (current status: ongoing). Retrospectively registered on 31 July 2017 ( https://clinicaltrials.gov/ct2/show/NCT03233984 ) because when the first participant was enrolled in this non-drug intervention, ClinicalTrials.gov was centered in therapeutic trials. The World Health Organization Trial Registration Data Set is in Additional file 1.

Perinatal Environmental Health Education Intervention to Reduce Exposure to Endocrine Disruptors: The PREVED Project.

Environmental health promotion interventions may reduce endocrine disruptor (ED) exposure. The PREVED (PREgnancy, preVention, Endocrine Disruptors) project was developed to improve knowledge, to enhance risk perception, and to change exposure behavior. Our objective was to present the phases of the PREVED project using the RE-AIM method. PREVED intervention consisted of three workshops during pregnancy. Reach, adoption, and implementation phases were assessed with qualitative studies. Efficacy study consisted of a three-arm randomized controlled trial (RCT) on 268 pregnant women: (i) control group (leaflet), (ii) intervention group in neutral location, (iii) intervention group in contextualized location. The main outcome was the percentage evolution of participants who reported consuming canned food. Secondary outcomes were evolution of psycho-social scores, evolution of ED presence in urine, and ED presence in colostrum. The intervention adoption was centered on upper-privileged women, but implementation assessment showed that key features (highly practical intervention) seemed to be carried out and had initiated some behavior changes. A total of 268 pregnant women participated in the intervention and 230 in a randomized controlled trial (control group: 86 and intervention groups: 172). We found no significant differences in consumption of canned food and in percentage of women having a decrease of bisphenol A or parabens in urine, but we found a significant increase in the evolution of risk perception score and overall psychosocial score in intervention groups (respectively: +15.73 control versus +21.03 intervention, p = 0.003 and +12.39 versus +16.20, p = 0.02). We found a significant difference in percentage of women with butylparaben detection between control group and intervention groups (13% versus 3%, p = 0.03). PREVED intervention is the first intervention research dedicated to perinatal environmental health education in France. By sharing know-how/experience in a positive non-alarmist approach, it improved risk perception, which is key to behavior change, aiming to reduce perinatal ED exposure. Including women in precarious situations remains a major issue.

How to Educate Pregnant Women about Endocrine Disruptors?

Background: Despite mediatization, only half of pregnant women are informed about endocrine disruptors (EDs). We wished to inquire about appropriate environmental health education procedures during pregnancy: Who, when, and how? Methods: The question stems from a comprehensive population health intervention research project. It includes qualitative studies aimed at constructing an educational program in environmental health and an accompanying assessment tool. The validation of a customized questionnaire (PREVED© for Pregnancy Prevention Endocrine Disruptors) about the knowledge, attitudes, and practices (KAP) of pregnant women regarding exposure to EDs was carried out in a quantitative study. Results: Health education by a prenatal professional with communication skills should take place as early as possible, during the preconception period or early pregnancy, as part of individual consultation or group workshops. In order to customize the discourse and to develop women's empowerment, concomitant presentation of the risks by the products used in each room and of previous solutions is recommended. Conclusion: Appropriate health education procedures on EDs should be done at every contact but taking the KAP of pregnant women into account first. We propose all educational actions should be accompanied by questioning of the KAP of pregnant women; for example, with questions from the PREVED© questionnaire.

Environmental Health Knowledge, Attitudes, and Practices of French Prenatal Professionals Working with a Socially Underprivileged Population: A Qualitative Study.

: Introduction: As environmental health knowledge of population is associated with social economic status, the objective of this study was to determine environmental health knowledge, attitudes, and practices of French prenatal professionals working with a socially underprivileged population. MATERIAL AND METHODS: A focus group with eleven prenatal professionals working with socially underprivileged population was carried out in France in 2015. Content analysis of verbatim explanation was conducted with choice-of-subject categories carried out according to the triangulation principle, and topic trees were generated and applied. RESULTS: The professionals have non-specialized experimental knowledge of emergent risks and were essentially preoccupied by infectious biological risks. In practice, however, they became increasingly cognizant of emergent risks. Their educational practices take cultural context into account but educational tools with imagination and affectivity have to be developed in order to reach socially underprivileged population. DISCUSSION: Professionals are often sensitized to the field of environmental health in their apprehension of polluted biological environments, which they associate with social contexts and behavioral factors. In this study, we recommend adapted training programs and updated professional guidelines in view of reducing prenatal emergent risk exposures and social inequalities of health.

Development and validation of an LC-MS/MS method for the simultaneous determination of bisphenol A and its chlorinated derivatives in adipose tissue.

Bisphenol A (BPA) and its chlorinated derivatives (Clx-BPA) are environmental pollutants exhibiting endocrine-disrupting (ED) properties suspected to be involved in the pathogenesis of hormone-dependent cancers, such as breast and prostate cancers. Due to their lipophilic properties, they may accumulate in adipose tissue which could therefore be a suitable matrix to assess long-term exposure to these compounds and relationships with the tumorigenesis of these cancers. An LC-MS/MS assay for the determination of BPA and Clx-BPA in adipose tissue samples was developed and fully validated according to current bioanalytical validation guidelines. Ionization was achieved using an electrospray source operating in the negative mode and quantification of target analytes was obtained in the multiple reaction monitoring mode. Both standard and quality control (QC) samples were prepared in blank adipose tissue samples. Linearity was demonstrated over the ranges 0.125 to 8.000 and 0.0125-0.8000 ng/mL for BPA and Clx-BPA, respectively. Accuracy and precision were demonstrated over the whole concentration range: intra and inter-day bias values were in the 85-114% range and imprecision of the method did not exceed 14%. Lower limits of quantification were validated using QCs at 0.1250 and 0.0125 ng/mL for BPA and Clx-BPA, respectively. Internal standard-corrected matrix effects were comparable in breast and prostate adipose tissues, demonstrating that this method could be used to reliably assay BPA and Clx-BPA in both tissues. The method was sensitive enough to determine BPA and Clx-BPA in breast adipose tissue obtained from women undergoing breast surgery, enabling identification of different patterns of exposure to these ED chemicals. The method enables the reliable quantification of BPA and Clx-BPA in adipose tissue and could be used to assess long-term exposure to these compounds and potential associations with hormone-dependent cancers.

Determinants of Risk Perception Related to Exposure to Endocrine Disruptors during Pregnancy: A Qualitative and Quantitative Study on French Women.

Endocrine disruptors (EDCs) are known as environmental exposure factors. However, they are rarely reported by health professionals in clinical practice, particularly during pregnancy, even though they are associated with many deleterious consequences. The objectives of this study were to estimate the risk perception of pregnant women related to EDC exposure and to evaluate its determinants. A qualitative study based on the Health Belief Model was carried out through interviews of pregnant women and focus group with perinatal, environmental health and prevention professionals in 2015 in the city of Poitiers, France. Then, determinants of risk perception were included in a questionnaire administered to 300 women in the perinatal period through a quantitative study. Scores were subsequently calculated. Perception of EDC risk was defined as perceived severity for different stages of the infant's development and perceived susceptibility to EDC exposure. The determinants reported in the qualitative study were: age, strong maternal figure, socio-professional category, level of knowledge, and involuntariness of exposure. Age and level of knowledge were confirmed in our statistical model as determinants. Mean score of EDC risk perception was 55.0 ± 18.3 on 100 points. Our study should guide healthcare providers when advising pregnant women about EDC and environmental exposure. Our score for perceived EDC risk and assessment of its known determinants may help to assess the impact and the relevance of prevention programs dedicated to reducing exposure to EDC during pregnancy.

Chlorinated and brominated bisphenol A derivatives: Synthesis, characterization and determination in water samples.

Bisphenol A (BPA) has been used in the plastics industry for several decades. During the treatment of drinking water with chlorine reagent, the formation of chlorinated derivatives of BPA (ClxBPA) but also bromoBPA and bromochloroBPA is to be expected. Some of these compounds are considered to have an estrogenic effect and could induce major risks for human health by targeting different organs and systems in the body. In this paper, we describe the synthesis of chloro- and bromobisphenol A (ClxBPA, BrxBPA, BrxClxBPA)and their analytical characterization. These derivatives could be used as analytical standards in LC-MS/MS or evaluated in in vitro biological tests for their potential as endocrine disruptors. In this study, we evaluated the presence of BPA, ClxBPA in a pilot study from water samples. Range values found for BPA, ClxBPA were respectively 2.8-4169.3 ng/L and 0.8-11.3 ng/L.

Endocrine Disruptors and Pregnancy: Knowledge, Attitudes and Prevention Behaviors of French Women.

Endocrine disrupting chemicals (EDC) are environmental exposure factors that are rarely reported in clinical practice, particularly during pregnancy. This study aimed to describe women's knowledge, attitudes and behaviors towards EDC exposure. A study was conducted in the French Department of Vienne between 2014 and 2016 and comprised semi-structured interviews with pregnant women, a focus group of professionals in perinatology and environmental health, and the administration of a psychosocial questionnaire comprising scores in 300 pregnant or in postpartum period women. The mean score of knowledge was 42.9 ± 9.8 out of 100 (from 13.5 to 75.7). Exposure attitude was determined by risk perception. Mean level of cues to action to reduce their EDC exposure was estimated at 56.9 ± 22.5 out of 100 (from 0 to 100). Anxiety was significantly increased after the questionnaire. Anxiety about EDC was associated with a high score of knowledge (OR = 2.30, 95% CI (1.12-4.71)) and with no pregnancy anxiety (OR = 0.57, 95% CI (0.34-0.95)). Our findings suggest that healthcare providers should consider pregnant women's knowledge and perceptions, possibilities of action, and be careful not to increase their anxiety when advising them about EDC and environmental exposure.

Ultrasensitive determination of bisphenol A and its chlorinated derivatives in urine using a high-throughput UPLC-MS/MS method.

Bisphenol A (BPA) is a well-known endocrine disruptor. Chlorinated derivatives of BPA (ClxBPA) may be formed by reaction of chlorine with BPA present in drinking water. ClxBPA exhibit a higher level of estrogenic activity than BPA. While many studies have reported detectable BPA concentrations in urine, only very few studies were conducted in regards to ClxBPA. Since ClxBPA are potentially more toxic, it is important to assess large-scale exposure of the general population. Indeed, in the field of environment health, large studies are required to assess exposure to pollutants at ultratrace concentrations; therefore, analytical methods have to be rapid and sensitive. This work intends to validate a highly sensitive and rapid analytical method suitable to evaluate BPA and ClxBPA exposures during large-scale biomonitoring studies. For that purpose, a method based on online solid-phase extraction coupled with isotope dilution ultrahigh - performance liquid chromatography-tandem mass spectrometry was developed and validated according to accepted guidelines. The matrix-matched calibration curve ranged from 0.25 to 16.0 ng mL(-1) and from 0.025 to 1.60 ng mL(-1) for BPA and ClxBPA, respectively. This method was precise (the intra- and inter-day coefficients of variation of quality control were <16.4%) and accurate (bias ranged from -4.0 to 16.8%). The limit of quantification was validated at 0.25 and 0.025 ng mL(-1), for BPA and ClxBPA, respectively. The limit of detection was estimated for each experiment performed. Finally, this method is rapid and sensitive enough to be carried out during biomonitoring studies of BPA and ClxBPA in human urine.

Antibacterial activities of oxyprenylated chalcones and napthtoquinone against Helicobacter pylori.

In this study, we synthesized and characterized the antibacterial activity of three naturally occurring oxyprenylated chalcones {xinjiachalcone A (1), (2E)-1-{2,6-dihydroxy-4-[(3-methylbut-2-enyl)oxy]phenyl}-3-(4-hydroxyphenyl)prop-2-en-1-one (2), (2E)-1-{2,6-dihydroxy-4-[(3-methylbut-2-enyl)oxy]phenyl}-3-phenylprop-2-en-1-one (3), and lawsone 2-isopentenyl ether (4)}. Using several strains of Helicobacter pylori, including clinical ones, minimum inhibitory concentration (MIC) values and bactericidal activities of these compounds were determined. Xinjiachalcone A (1), active principle of Glycyrrhiza inflata Batalin, was the most effective compound, showing both a low MIC and a strong bactericidal activity against H. pylori. This study suggests that these compounds represent potential natural molecules for the prevention and treatment of H. pylori associated diseases.