Fecal calprotectin (fCal) is a non-invasive biomarker used in the diagnosis, prognostic evaluation, and monitoring of treatment response in inflammatory bowel diseases. Turbidimetric assays provide an automated alternative to the ELISA method, enabling faster and large-scale determination of fCal levels. The objective of this study was to evaluate the analytical performance of the Bühlmann fCAL® turbo assay, with particular emphasis on reagent and analyte stability.Patients’ samples, as well as internal and external quality controls from Montauban Hospital, were analyzed using the Bühlmann fCAL® turbo kit on a Cobas c503 analyzer.This study demonstrated that this immunoturbidimetric technique exhibits appropriate analytical performance. The on-board reagent kit stability was found to extend to at least 7 months, while calibration remained stable for up to 10 weeks. An increase in fCal concentrations was observed following 15 days of storage at +4°C. The rational method validation strategy proposed in this study could facilitate the implementation of this technique in clinical laboratories, particularly in small to medium-sized centers.The Bühlmann fCAL® turbo assay is a reliable and efficient turbidimetric method for rapid fCal measurement, offering extended reagent stability, robust analytical performance, and reduced costs.

Psoriasis disease (PsD) is an immune-mediated inflammatory disease (IMID) associated with comorbidities, as cardiovascular diseases (CVD). Several factors have been identified as trigger for CVD in PsD. As for other IMID, we propose to investigate the role of vitamin B6 (B6) in hyperhomocystinemia and cardiovascular risk management in order to individualize PsD care psoriasis. B6 deficiencies are described for other IMID such as rheumatoid arthritis. An increased consumption of B6 on the tryptophan/kynurenine pathway of T-lymphocytes is one of the principal mechanism leading to a decrease in B6 status. Then low B6 status is a hallmark of disease activity but is also associated with impaired clearance of homocysteine, which accumulating and increasing the risk of CVD. Management of B6 deficiency still needs to be explored in PsD care management. There is limited data available on the benefits of repletion, even for other IMID. Repletion should be individualized based on the patient's B6 status but also on PsD severity. Indeed, aiming physiological B6 status could be relevant in order to prevent hyperhomocysteinemia and, consequently, CVD.

Compounded oral suspensions of spironolactone are commonly used in pediatric patients with heart conditions. However, due to its poor solubility in water, spironolactone suspensions may lead to sedimentation, compromising homogeneity and dose accuracy. This study aimed to assess dose accuracy of spironolactone suspension in critical care settings. Over one month in 2020 and 2022, samples prepared by nurses in Pediatric (PICU) and Neonatal Intensive Care Units (NICU) were collected and analysed using an HPLC/UV validated method to evaluate dose deviations from target. Educational interventions on suspension preparation were conducted between the two periods. In 2020, only 28% (NICU) and 0% (PICU) of samples met target dose with greater variability in PICU. After educational intervention from the pharmacy teams, in 2022 improvements were observed, but underdosing and variability remained significant, reflecting ongoing challenges in suspension homogeneity. These findings highlight the need for a more stable pediatric formulation to enhance patient safety and treatment effectiveness.

The pediatric population often faces challenges in accessing appropriate medication formulations, particularly for circumstances like congenital heart disease requiring spironolactone therapy. This study aimed to optimize the pharmaceutical formulation of oral suspension spironolactone for pediatric use and assess its stability. A formulation with 0.2 % xanthan gum in InOrpha® was developed, showing improved stability and reduced sedimentation. Analytical method validation confirmed accuracy and precision for spironolactone quantification, while forced degradation studies ensured stability-indicating capability. Stability assessments demonstrated the oral suspension's chemical, physical, and microbiological stability for up to 135 days pre-bottle opening and 37 days post-opening under varied storage conditions. This study provides crucial insights into enhancing spironolactone formulation for pediatric patients. Further research is needed to assess pharmacokinetic parameters such as bioavailability and pharmacodynamics to fully ascertain its efficacy in pediatric populations.

ObjectivesVibrio cholerae non-O1/non-O139 (NOVC) bacteremia is infrequently reported in Western countries and is associated with unfavorable outcome.Patient/MethodWe describe here the case of a diabetic patient with hepatic cytolysis and NOVC bacteremia following an episode of diarrhea.ResultThe patient was paucisymptomatic and had a favorable resolution with oral ciprofloxacin.ConclusionNOVC should be systematically sought in stool samples, particularly in immunocompromised patients, due to an increased risk of infection occurrence.

Amid the early 2020 SARS-CoV-2 crisis, severe hand sanitizer shortages led to OMS local production recommendations, inviting a diverse array of alcohol producers to contribute. However, not all followed mandatory controls for API-grade alcohol. We conducted a study to ensure the safety of the received alcohols, focusing on methanol and acetaldehyde levels. All samples were well below Ph. Eur guidelines, affirming their safety for use. Furthermore, no additional impurities were detected, reinforcing the quality and safety of the assessed hand sanitizers. Our findings, amidst the scarcity of the SARS-CoV-2 era, highlight the importance of rigorous safety assessments during local hand sanitizer production.

Many studies suggest that the potential impact of bisphenol S (BPS) as an endocrine disruptor is comparable to that of bisphenol A (BPA). However, in vitro-to-in vivo and from animal to human extrapolations require knowledge of the plasma free fraction of the active endocrine compounds. The present study aimed to characterise BPA and BPS binding to plasma proteins both in humans and different animal species. The plasma protein binding of BPA and BPS was assessed by equilibrium dialysis in plasma from adult female mice, rats, monkeys, early and late pregnant women as well as paired cord blood, early and late pregnant sheep and foetal sheep. The fraction of free BPA was independent of plasma concentrations and ranged between 4% and 7% in adults. This fraction was 2 to 3.5 times lower than that of BPS in all species except sheep, ranging from 3% to 20%. Plasma binding of BPA and BPS was not affected by the stage of pregnancy, BPA and BPS free fractions representing about 4% and 9% during early and late human pregnancy, respectively. These fractions were lower than the free fractions of BPA (7%) and BPS (12%) in cord blood. Our results suggest that similarly to BPA, BPS is extensively bound to proteins, mainly albumin. The higher fraction of free BPS compared to BPA may have implications for human exposure assessment since BPS free plasma concentrations are expected to be 2 to 3.5 times higher than that of BPA for similar plasma concentration.

Background The European Society of Cardiology (ESC) guidelines recommend a dynamic (0-1h) cardiac troponin (cTn) determination for non-ST elevation myocardial infarction diagnosis. For patients with low cTn levels, a discharge from emergency can be considered. Nevertheless, cTn cutoffs for discharge are lower than the limits of quantification proposed by laboratory reagent suppliers. Aim Validate cTn assay on the Elecsys STAT kit. Materials and methods Precision, trueness, repeatability and within-laboratory variability were calculated from internal quality control and plasma pooled at 5.78 and 10.73 ng/L. Accuracy was calculated from external quality control. Uncertainty of measurement was calculated from (i) the uncertainty of the standard and control values and (ii) by precision from pooled plasma. Distribution of precision results from pooled plasma has been evaluated by bootstrap simulations. Dilution linearity tests with patient plasma were performed to evaluate the method for values near 5 ng/L. Results Precision and trueness ranged from 1.35 to 4.45% and from 0.14 to −3.74%, respectively. Accuracy results ranged from 101.40 to 104.90%. Within laboratory variability was 2.91%. Uncertainty ranged from 3.66% to 19.90% for higher (2188) to lower values (5.78 ng/L). Bootstrap simulations allowed utilization of precision data from pooled plasma to evaluate cTn assay. The method was linear from 4.48 to 39.80 ng/L. A linear regression model best described the data. Conclusion Elecsys STAT method provides accurate cTn results, including patients with cTn results categorizing them as ‘rule-out’ in the ESC guidelines.

La prévention en santé comprend un apport alimentaire adéquat en macronutriments (glucides, lipides, protides) et en micronutriments (vitamines, oligoéléments). Les micronutriments jouent un rôle essentiel comme cofacteurs ou agents réducteurs dans la quasi-totalité des réactions biochimiques du métabolisme cellulaire. L’objectif de cette revue est de proposer une mise à jour des connaissances du métabolisme des principaux micronutriments, vitamines hydrosolubles (B1, B6, B9, B12, C), liposolubles (A, E, D) et oligoéléments (fer, zinc, cuivre, sélénium) avec une attention particulière sur l’impact de l’inflammation. Cette revue a également pour but de positionner le statut des micronutriments dans des groupes à risques de carence émergents (MICI, chirurgie bariatrique, résection intestinale, Covid) et d’en dégager les principaux messages de mise en pratique. Les biomarqueurs d’un excès en micronutriments sont généralement bien moins caractérisés que ceux d’une carence en particulier en termes de liens avec des symptômes fonctionnels ou cliniques de toxicité. La prévention des carences en l’absence de pathologie, repose sur une alimentation variée et équilibrée qui couvre facilement l’ensemble des besoins. Une prévention par complément micronutritionnel peut se trouver justifiée de façon temporaire ou prolongée dans des situations physiopathologiques particulières (régimes particuliers, interactions médicamenteuses, pathologies).

Health prevention includes an adequate dietary intake of macronutrients (carbohydrates, lipids, proteins) and micronutrients (vitamins, trace elements). Micronutrients play an essential role as co-factors or reducing agents in almost all of the biochemical reactions of cellular metabolism. The objective of this review is to provide an update of knowledge of the metabolism of the main micronutrients such as water-soluble (B1, B6, B9, B12, C) and fat-soluble (A, E, D) vitamins and trace elements (iron, zinc, copper, selenium) with a particular attention to their impact of inflammation. The aim of the review is also to define the status of micronutrients in groups at risk of emerging deficiency (inflammation bowel disease, bariatric surgery, bowel resection, COVID) and to draw out the main messages for putting them into practice. Biomarkers of micronutrient excess are generally less well characterized than those of deficiency, particularly in terms of links to functional or clinical symptoms of toxicity. The prevention of deficiencies in the absence of pathology, is based on a diet varied and balanced that easily covers all needs. Prevention by micronutritional supplements may be justified on a temporary or prolonged basis in particular pathophysiological situations (special diets, drug interactions, pathologies).(c) 2023 Societe francophone nutrition clinique et metabolisme (SFNCM). Published by Elsevier Masson SAS. All rights reserved.

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.

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.

Metsu, Davida,b; Aquilina, Christianc; Delobel, Pierred,e; Gandia, Peggya,b; Savagner, Frédériquef,g; Raymond, Stéphaniee,h; Caron, Philippeg,i; Martin-Blondel, Guillaumed,eAuthor Information

Dolutegravir therapeutic drug monitoring (TDM) could be improved by measuring the unbound dolutegravir plasma concentration (Cu), particularly in patients experiencing virological failure or toxicity despite achieving appropriate DTG total plasma concentrations. Equilibrium dialysis (ED) is the gold standard to measure Cu, but ED is time consuming, precluding its use in clinical practice. In contrast, ultrafiltration is applicable to TDM, but is sensitive to numerous analytical conditions. In order to evaluate measurements of Cu by ultrafiltration, ultrafiltration conditions were validated by comparison with ED. DTG concentrations were measured by LC–MS/MS. Three ultrafiltration factors (temperature, duration and relative centrifugal force [RCF]) were evaluated and compared to ED (25/37 °C), using a design of experiment strategy. Temperature was found to influence Cu results by ED ( p = 0.036) and UF ( p = 0.002) when results were analysed with ANOVA. Relative centrifugal force (2000 g) and time (20 min) interacted to influence Cu ( p = 0.006), while individually they did not influence Cu ( p = 0.88 and p = 0.42 for RCF and time). Ultrafiltration conditions which yielded the most comparable results to ED were 37 °C, 1000 g for 20 min. Ultrafiltration results greatly depended on analytical conditions, confirming the need to validate the method by comparison with ED in order to correctly interpret DTG Cu.

Integrase inhibitors are increasingly used in combined antiretroviral therapy (ART) because of their high efficacy in inhibiting HIV replication and favourable tolerance profile. Elvitegravir is used with the pharmacokinetic enhancer cobicistat, associated with tenofovir and emtricitabine in a single-tablet regimen [1]. Very few data are available regarding the diffusion of elvitegravir in the central nervous system (CNS) [2,3]. Herein, we report a case of HIV-associated neurocognitive disorder (HAND) on elvitegravir-based treatment with virological escape in the CNS and the emergence of the elvitegravir-resistance mutation T66I in the cerebrospinal fluid (CSF). A 45-year-old HIV-1 infected woman presented in September 2017 with rapidly progressive cognitive impairment and headache for a few weeks. Her HIV-1 subtype-B infection was discovered in 1993. She was negative for HBV and HCV. Her CD4+ T cell count nadir was 130 cells/μl. She had a previous history of HIV encephalitis in 2014, following an 8-year interruption of ART. Her history of ART, viral load and drug-resistance mutation patterns is summarized in Fig. 1a. Since July 2016, she was successfully receiving tenofovir disoproxil fumarate [245 mg daily (q.d.)], emtricitabine (200 mg q.d.) and dolutegravir (50 mg q.d.), which had been switched in June 2017 to a single-tablet regimen of tenofovir alafenamide (10 mg q.d.), emtricitabine (200 mg q.d.) and cobicistat-boosted elvitegravir (150/150 mg q.d.). Plasma HIV-1 RNA was less than 1.3 log copies/ml at this time. She declared perfect adherence to treatment.Fig. 1: (a) HIV-infection course.(b) Brain MRI. Axial brain MRI with contrast-enhanced FLAIR sequence in September 2017 showing mild cerebral atrophy and periventricular white matter lesions compatible with HIV-encephalitis, and leptomeningeal enhancement associated with meningitis. 3TC, lamivudine; DRV/r, ritonavir-boosted darunavir; DTG, dolutegravir; Env, envelope gene; EVG/c, cobicistat-boosted elvitegravir; FTC, emtricitabine; IN, integrase gene; MVC, maraviroc; NVP, nevirapine; PR, protease gene; R5, CCR5-using viruses; RT, reverse transcriptase gene; TAF, tenofovir alafenamide; TDF, tenofovir disoproxil fumarate; WT, wild-type; ZDV, zidovudine. Black square, measure in peripheral blood; open diamond, measure in CSF.At admission in September 2017, HIV-1 RNA was 2.7 log copies/ml in plasma and 4.3 log copies/ml in CSF. CSF examination revealed 70 WBC/μl (all of whom were mononuclear cells), increased protein level of 96 mg/dl and normal glucose level of 2.87 mmol/l. All investigations looking for bacteria, mycobacteria, fungi and other viruses remained negative. Genotypic assessment of HIV-1 for drug resistance revealed M184 V mutation in the reverse transcriptase gene, both in blood and CSF. T66I mutation associated with resistance to elvitegravir was found in CSF. The integrase gene could not be amplified in blood samples due to low viral load. Phenotypic tropism assessment in blood and CSF revealed only CCR5-using viruses. Drug dosages by LC-MS/MS revealed tenofovir concentrations of 17 ng/ml in plasma (time, 24 h) and 2.77 ng/ml in CSF (time, 8 h); emtricitabine concentrations of 100 ng/ml in plasma (time, 24 h) and 140 ng/ml in CSF (time, 8 h); and elvitegravir of 540 ng/ml (total concentration) and 0.425 ng/ml (unbounded concentration, measure adapted from [4]) in plasma (time, 24 h), and 15.7 ng/mL (total concentration) in CSF (time, 8 h). Brain MRI was compatible with HIV-encephalitis (Fig. 1b). ART was changed in September 2017 to tenofovir disoproxil fumarate (245 mg q.d.), emtricitabine (200 mg q.d.), ritonavir-boosted darunavir [600/100 mg twice daily (b.i.d.)] and maraviroc (300 mg b.i.d.). Zidovudine (300 mg b.i.d.) was added in January 2018 because of incomplete virological control in CSF (HIV-1 RNA of 2.86 log copies/mL in CSF vs. 1.75 log copies/ml in plasma). In April 2018, both plasma and CSF HIV-1 RNA were less than 1.47 log copies/ml. All neurological symptoms progressively disappeared. Herein, we report the first case of virological escape in the CNS on elvitegravir-based treatment associated with clinical HAND and the selection of T66I elvitegravir-resistance mutation in the CSF. Some factors could have contributed to this escape: this participant probably harboured HIV-1 strains with particular neurotropism, as she already had a past history of HIV-encephalitis; pre-existing M184 V emtricitabine-resistance mutation impaired the potency of the antiretroviral regimen. Virological escape in the CNS occurred 3 months after switching the integrase inhibitor from dolutegravir to elvitegravir, suggesting lower diffusion in the CNS and/or weaker virological potency of elvitegravir than dolutegravir in this case. Trough plasma total concentrations of antiretroviral drugs were adequate [5]. Total concentration (Ct) measured during therapeutic drug monitoring usually represents a good substitute for unbound concentration (Cu). Through its ability to cross cell membranes to achieve pharmacodynamic activity, the Cu is considered as the pharmacologically active form [6]. In our case, high level of CSF-protein may have led to an increase in elvitegravir-bound concentration and therefore in the Ct, without modification of Cu. At equilibrium, Cu in the CSF is less than or equal to plasma Cu. Hence, on the basis of plasma Cu, it may be assumed that the Cu in the CSF was lower than the elvitegravir IC50 (3.9 ng/ml) [7]. This low exposure could explain the compartmentalized virological failure with emergence of elvitegravir-resistance mutation. The risk of virological escape in the CNS on elvitegravir-based regimen should be further estimated, in an era in which integrase inhibitors are becoming the main third agent of combined ART. Acknowledgements Conflicts of interest There are no conflicts of interest.

Teicoplanin is often used in Enterococcus faecalis infective endocarditis as a relay in case of penicillin side effects, or in outpatients. We assessed the efficacy of teicoplanin used as continuation therapy after initial standard treatment of E. faecalis endocarditis.

Hepatitis E virus (HEV) infection causes chronic hepatitis in solid organ transplant (SOT) recipients. Antiviral therapy consists of three months of ribavirin, although response rates are not optimal. We characterized plasma HEV kinetic patterns in 41 SOT patients during ribavirin therapy. After a median pharmacological delay of three (range: 0–21) days, plasma HEV declined from a median baseline level of 6.12 (3.53–7.45) log copies/mL in four viral kinetic patterns: (i) monophasic (n = 18), (ii) biphasic (n = 13), (iii) triphasic (n = 8), and (iv) flat-partial response (n = 2). The mean plasma HEV half-life was estimated to be 2.0 ± 0.96 days. Twenty-five patients (61%) had a sustained virological response (SVR) 24 weeks after completion of therapy. Viral kinetic patterns (i)–(iii) were not associated with baseline characteristics or outcome of therapy. A flat-partial response was associated with treatment failure. All patients with a log concentration decrease of plasma HEV at day seven of >15% from baseline achieved SVR. In conclusion, viral kinetic modeling of plasma HEV under ribavirin therapy showed, for the first time, four distinct kinetic profiles, a median pharmacologic delay of three days, and an estimated HEV half-life of two days. Viral kinetic patterns were not associated with response to therapy, with the exception of a flat-partial response.