Extreme hyperferritinemia in the setting of acute myeloid leukaemia: a case report of hemophagocytic lymphohistiocytosis.

Introduction: Major hyperferritinemia is a rare feature in clinical laboratories associated with a wide variety of disorders, including hemophagocytic lymphohistiocytosis (HLH). The diagnosis of HLH is based on clinical and biological criteria, such as those proposed by the Histiocyte Society. However, several of these criteria are not relevant in the specific setting of hematologic malignancies. Materials and methods: A 69-year-old male was treated for an acute myeloid leukaemia. On day 15 after the start of chemotherapy, he developed severe sepsis with high fever, low blood pressure and hepatosplenomegaly. Results: Blood tests were marked by extreme hyperferritinemia (191,000 µg/L, reference range: 26-388 µg/L) with increased C-reactive protein (87.0 mg/L) and procalcitonin (1.94 µg/L) and aspartate aminotransferase (499 U/L 37 °C) in the setting of chemotherapy-induced aplasia. This unusual extreme ferritinemia led to suspect HLH triggered by an invasive infection. Under intensive treatment, the clinical status improved and ferritin levels significantly decreased. Conclusions: The diagnosis of HLH is usually based on clinical and biological criteria, mainly fever, splenomegaly, cytopenias, hypertriglyceridemia, hypofibrinogenemia, hemophagocytosis and hyperferritinemia. In this patient, the diagnosis of HLH was challenging because several criteria, such as hypertriglyceridemia, hemophagocytosis and hypofibrinogenemia, were absent. In addition, some criteria of HLH are not relevant in the setting of hematologic malignancy, in which fever, splenomegaly, cytopenias and elevated lactate dehydrogenase are commonly observed independently of HLH. This unusual case of extremely high ferritinemia emphasizes the important weight of the ferritin level for the diagnosis of HLH in adult patients in the setting of hematologic malignancies

Apolipoprotein B is regulated by gonadotropins and constitutes a predictive biomarker of IVF outcomes

International audienceAbstractBackgroundFollicular fluid (FF) is an important micro-environment influencing oocyte growth, its development competence, and embryo viability. The FF content analysis allows to identify new relevant biomarkers, which could be predictive of in vitro fertilization (IVF) outcomes. Inside ovarian follicle, the amount of FF components from granulosa cells (GC) secretion, could be regulated by gonadotropins, which play a major role in follicle development.MethodsThis prospective study included 61 female undergoing IVF or Intra-cytoplasmic sperm injection (ICSI) procedure. Apolipoprotein B (APOB) concentrations in follicular fluid and APOB gene and protein expression in granulosa cells from reproductively aged women undergoing an in vitro fertilization program were measured. The statistical analyses were performed according to a quartile model based on the amount of APOB level found in FF.ResultsAmounts of APOB were detected in human FF samples (mean ± SD: 244.6 ± 185.9 ng/ml). The odds of obtaining an oocyte in the follicle and a fertilized oocyte increased significantly when APOB level in FF was higher than 112 ng/ml [i.e., including in Quartile Q 2, Q3 and Q4] (p = 0.001; p < 0.001, respectively). The probabilities of obtaining an embryo and a top quality embryo on day 2, were significantly higher if APOB levels were within the ranges of 112 and 330 ng/ml (i.e. in Q2 and Q3) or 112 and 230 ng/ml (i.e. in Q2), respectively (p < 0.001; p = 0.047, respectively). In addition, our experiments in vitro indicated that APOB gene and protein expression, along with APOB content into culture were significantly under-expressed in GC upon stimulation with gonadotropins (follicular stimulating hormone: FSH and/or human chorionic gonadotropin: hCG).ConclusionWe are reporting a positive and statistically significant associations between APOB and oocyte retrieval, oocyte fertilization, and embryo quality. Using an experimental study component, the authors report significant reduced APOB expression and content for luteinized granulosa cells cultured in the presence of gonadotropins

Complete exon sequencing of all known Usher syndrome genes greatly improves molecular diagnosis

<p>Abstract</p> <p>Background</p> <p>Usher syndrome (USH) combines sensorineural deafness with blindness. It is inherited in an autosomal recessive mode. Early diagnosis is critical for adapted educational and patient management choices, and for genetic counseling. To date, nine causative genes have been identified for the three clinical subtypes (USH1, USH2 and USH3). Current diagnostic strategies make use of a genotyping microarray that is based on the previously reported mutations. The purpose of this study was to design a more accurate molecular diagnosis tool.</p> <p>Methods</p> <p>We sequenced the 366 coding exons and flanking regions of the nine known USH genes, in 54 USH patients (27 USH1, 21 USH2 and 6 USH3).</p> <p>Results</p> <p>Biallelic mutations were detected in 39 patients (72%) and monoallelic mutations in an additional 10 patients (18.5%). In addition to biallelic mutations in one of the USH genes, presumably pathogenic mutations in another USH gene were detected in seven patients (13%), and another patient carried monoallelic mutations in three different USH genes. Notably, none of the USH3 patients carried detectable mutations in the only known USH3 gene, whereas they all carried mutations in USH2 genes. Most importantly, the currently used microarray would have detected only 30 of the 81 different mutations that we found, of which 39 (48%) were novel.</p> <p>Conclusions</p> <p>Based on these results, complete exon sequencing of the currently known USH genes stands as a definite improvement for molecular diagnosis of this disease, which is of utmost importance in the perspective of gene therapy.</p

Composition en phospholipides et sphingolipides des lipoprotéines de haute densité (HDL) chez les diabétiques de type 1

Objectif. En raison de l importance des phospholipides (PL) et sphingolipides (SPL) dans la fonctionnalité des HDL, nous avons recherché une modification de la composition en PL et SPL des HDL chez 43 patients DT1 et 36 sujets témoins à l aide d une analyse par chromatographie liquide couplée à la spectrométrie de masse en tandem. Résultats. Les HDL étaient appauvries en sphingosine-1-phosphate au cours du DT1 (581 +- 165 versus 659 +- 148 nmol/l chez les sujets DT1 et témoins respectivement, p = 0,033). La proportion de PL par rapport à l ensemble des constituants des HDL était augmentée chez les sujets DT1 (+ 9 %, p < 0,001). L activité PLTP était augmentée de 21 % au cours du DT1 (p = 0,001). Conclusion. Ces modifications sont susceptibles de contribuer à l altération de la fonctionnalité des HDL observée au cours du DT1DIJON-BU Médecine Pharmacie (212312103) / SudocSudocFranceF

Développement de méthodes de référence pour les biomarqueurs du bilan lipidique (application au contrôle qualité en biologie clinique)

En biologie clinique, il est indispensable de disposer de mesures fiables et comparables dans le temps et entre les laboratoires afin de permettre un dépistage et un suivi appropriés des patients. Pour cela, il est indispensable d établir leur traçabilité métrologique aux unités du système international notamment par des méthodes de référence primaires ou des matériaux de référence certifiés (MRC) d ordre supérieur. Ces travaux de thèse ont consisté à développer et valider des méthodes de référence pour le cholestérol total, les triglycérides, le HDL-cholestérol et le LDL-cholestérol. Leur valeur ajoutée par rapport à une valeur consensuelle a été démontrée lors d évaluations externes de la qualité. Elles ont également permis de certifier un MRC qui pourra être utilisé pour le contrôle qualité et/ou l étalonnage des méthodes de routine. Nous avons montré que le MRC était commutable pour la plupart des méthodes de routine pour les différents biomarqueurs, ce qui a permis de l utiliser pour évaluer leur justesse. Les méthodes de routine avaient généralement tendance à sous-estimer la concentration en triglycérides (en particulier aux valeurs basses) et à surestimer nettement la concentration de cholestérol total et de LDL-cholestérol (en particulier aux concentrations proches du seuil de décision clinique), ce qui se traduit par une augmentation du nombre de faux-positifs (patients traités à tort). Une approche de correction de non commutabilité a également été proposée afin de permettre l utilisation de matériaux non commutables pour évaluer la justesse. Pour conclure, ces travaux ont démontré l importance de disposer de méthodes de référence ainsi que de MRC commutablesReliable measurements in medical biology are essential for early screening and appropriate follow-up of patients. Ensuring metrological traceability of clinical measurements to higher order reference methods or certified reference materials enables to obtain comparable results over time and between different laboratories that could use different methods to quantify the same biomarker.In this study, reference methods were developed and validated for lipid profile biomarkers (total cholesterol, triglycerides, HDL-C, and LDL-C). Their value added in proficiency testing schemes was demonstrated against consensus mean. They were also used to characterize a certified reference material (CRM) that may be used both as quality control and/or calibrator of field methods. The CRM was shown to be commutable for most field methods and lipid profile biomarkers, which proved it was suitable to assess trueness. Results of our multicenter study showed that field methods tend to underestimate triglycerides (particularly at low concentrations) and overestimate total cholesterol and LDL-C (especially around the clinical threshold), resulting in false positives and significant patient misclassifications. An approach of non-commutanility correction was also presented to allow trueness assessment with non-commutable samples. In conclusion, this work highlights the importance of using reference methods and also commutable CRM to rigorously assess accuracy of field methods used in clinical laboratoriesDIJON-BU Doc.électronique (212319901) / SudocSudocFranceF

Evaluation of the new restandardized 25-hydroxyvitamin D assay on the iSYS platform

IF 2.434International audienceBACKGROUND:25-hydroxyvitamin D [25(OH)D] is the most reliable biomarker of vitamin D status, but until now 25(OH)D assays have suffered from inter-laboratory and inter-assay discrepancies. In the setting of the international Vitamin D Standardization Program, Immunodiagnostic Systems (IDS) recently reformulated and restandardized the 25(OH)D immunoassay available on the automated iSYS platform. In the present study, we evaluated this new generation of the 25(OH)D immunoassay (IS-2500).METHODS:Repeatability and within-laboratory imprecision were verified according to the Clinical and Laboratory Standards Institute EP15-A3. Results from the sera of 63 patients were compared with those obtained with the previous iSYS method (IS-2700S) using Passing-Bablok and Bland-Altman analysis. The prevalence and bias-adjusted kappa (PABAK) coefficient was calculated to assess the agreement of vitamin D status provided by the two iSYS immunoassays. Fourteen Vitamin D External Quality Assessment Scheme (DEQAS) samples were used to evaluate inaccuracy.RESULTS:Using the EP15-A3 protocol, repeatability and within-laboratory imprecision obtained with the new iSYS method were lower than 6% and 8%, respectively. These results are consistent with the manufacturer's claims. In more adverse conditions (50 measurements over 15days with multiple calibrations), the within-laboratory imprecision was 14.8% (39nmol/L) and 7.7% (155nmol/L). 25(OH)D concentrations measured with the new assay showed a strong correlation with those provided by the previous version (r=0.969, p<0.0001). The Passing-Bablok regression equation was as follows: new assay=1.079 x (previous assay) - 3.6nmol/L. The PABAK coefficient of 0.810 reflected almost perfect agreement between the two immunoassays to classify patients according to their vitamin D status (85.7% of agreement). Using DEQAS samples, the mean inaccuracy bias was lower than 5% when the new iSYS method was compared with LC-MS/MS methods and the NIST reference measurement procedure.CONCLUSION:The new generation of the iSYS immunoassay evaluated in this study meets requirements for routinely measuring 25(OH)D levels in clinical laboratories.Copyright © 2017. Published by Elsevier Inc

Assessment of folate status in obese patients: Should we measure folate in serum or in red blood cells?

BACKGROUND: Recent studies revealed that obesity is associated with decreased serum but at the same time increased red blood cell (RBC) folate concentrations compared with lean subjects, thus casting doubt upon the agreement between serum and RBC folate measurements for assessing folate status. This work aimed to determine whether these two metrics lead to the same classification of folate status in obese patients. METHODS: RBC and serum folate concentrations were measured with a chemiluminescent immunoassay in 263 adults with body mass index >/=30 kg/m2 and without previous bariatric surgery. Among them, 68.1 % were eligible for bariatric surgery. Each serum and RBC folate result was classified as deficient or not according to thresholds recommended by the kit manufacturer (model A) or by the World Health Organization (model B). The agreement between serum and RBC folate results was evaluated using the proportion of overall agreement and the prevalence-adjusted bias-adjusted kappa (PABAK) statistics. RESULTS: The overall percentage agreements between serum and RBC measurements were 91.6 % (95 % CI 87.6-94.7 %) and 92.4 % (95 % CI 88.5-95.3 %) with PABAK coefficients of 0.87 (95 % CI 0.82-0.93) and 0.88 (95 % CI 0.83-0.94) in the models A and B, respectively, corresponding to almost perfect agreement. The same was true in the subgroup of patients eligible for bariatric surgery. Gender, age, and BMI did not influence the quality of agreement between the two parameters. CONCLUSIONS: The present study showed that folate measurements in serum and in RBC display similar performances to assess folate status in obese patients

Interférences des traitements par biotine avec les immunodosages : il est temps de trouver une solution à long terme !

IF 0.908 (2017)Lettre à l'éditeur ("La Presse Médicale" vol. 47 n°1)https://www.sciencedirect.com/science/article/pii/S075549821730461X?via%3Dihu

Effects of Transcranial Magnetic Stimulation on the Hypothalamic-Pituitary Axis in Depression: Results of a Pilot Study

International audienceSome studies have reported that repetitive transcranial magnetic stimulation (rTMS) applied to the dorsolateral prefrontal cortex (DLPFC) is able to induce changes in the hypothalamic-pituitary axis in subjects with major depression. The causes of these neuroendocrine effects are unknown and deserve to be studied. The authors monitored neuroendocrine hormones in 15 subjects with major depression treated by 1-Hz rTMS on the right DLPFC and explored a correlation with mood improvement. Unlike previous studies, no changes in serum cortisol, prolactin, and thyroid hormone levels were found. However, the authors did observe short-term changes in growth hormone levels in nonresponsive subjects