Overview of the in vitro stability of commonly measured vitamins and carotenoids in whole blood

Background The pre-analytical stabilities of vitamins A, E, K, B1, B2, B6, B12, C, carotenoids and folates in whole blood were studied. The aim of this work was to provide clear and workable pre-analytical procedures specifying optimal delay before freezing for laboratories which perform themselves such analyses or which receive and transfer such specimens to referral laboratories.Methods The stability of vitamins was studied in whole blood at room temperature after light exposure up to 24h (vitamin C), 48h (vitamins A, E, B1, B2, B6 and carotenoids) and 72h (vitamins K, B12, red blood cell (RBC) and serum folates). Vitamin C stability after baseline acidification was analysed up to 48h. Changes observed were compared to a clinical cut-off defined as total change limit based on a combination of analytical performance and within-subject variation.Results Clinically and statistically significant changes occurred only in vitamins C (-22.5%), B6 (+9.9%) and serum folates (-16.8%) concentrations after 6, 24 and 48h, respectively. Vitamins A, E, K, B1, B2, B12, RBC folates and carotenoids showed good stability up to 48 or 72h. Vitamin C in acidified serum conserved at room temperature appeared unstable. The optimal condition for acidified vitamin C conservation was at less than -20?.Conclusion The majority of vitamins remain stable for up to 48h. Vitamin C quantification requires serum acidification followed by freezing as soon as possible. Freezing, respectively, within 12h and 24h for determination of plasma vitamin B6 and serum folates concentrations is recommended

What is the care pathway of patients who undergo thyroid surgery in France and its potential pitfalls? A national cohort

International audienceContext: The rate of thyroid cancer is increasing in France, as well as concerns about overdiagnosis and treatment.Objectives: To examine the care pathway of patients who undergo thyroid surgery in France and detect potential pitfalls.Design: A large observational study based on medical reimbursements, 2009–2011.Setting: Data from the Sniiram (National Health Insurance Information System).Patients: Patients with thyroid surgery in 2010, classified into 4 groups: thyroid cancer, benign nodule, goitre or multiple nodules, other (hyperthyroidism, head–neck cancer).Main outcome: measures Medical investigations during, prior and after thyroidectomy.Results: A total of 35 367 patients underwent surgery (mean age 51 years, 80% women): 17% had a reported diagnosis of thyroid cancer, 20% benign nodule, 38% goitre or multiple nodules and 25% another diagnosis. The ratio of thyroidectomies with cancer over thyroidectomies with benign nodule was 0.8 and varied across regions. In the year preceding surgery, 82% of patients had an investigation by thyroid ultrasonography, 21% thyroid scintigraphy, 34% fine-needle aspiration cytology, 40% serum calcitonin assay and 54% serum calcium assay. In the following year, all patients with total thyroidectomy and 44% of patients with partial thyroidectomy and a diagnosis of benign nodule were taking thyroid hormone therapy. 100 patients had been reoperated for a compressive haematoma and 63 died during the first month, half of whom had been operated for cancer. Mean rates of recurrent laryngeal nerve injury and hypocalcaemia (requiring blood tests plus treatments within 4–12 months) were estimated at 1.5% and 3.4%, respectively, and were higher in the cancer group (2.3% and 5.7%).Conclusions: This almost nationwide study demonstrates the suboptimal management of patients prior to thyroidectomy in France. It suggests overdiagnosis and potential harms to patients, and calls for a review of the relevance of thyroidectomy, particularly with regard to microcancers

Accuracies of serum and fecal S100 proteins (calprotectin and calgranulin C) to predict the response to TNF antagonists in patients with Crohn's disease

International audienceBACKGROUND: Calprotectin and S100A12 (calgranulin C) are markers of gut inflammation. The aim was to compare the usefulness of serum and fecal calprotectin (fCal) and S100A12 in assessing the response to anti-TNF and in predicting relapse under maintenance therapy in Crohn's diseases (CD). METHODS: Thirty-two consecutive patients with CD were treated with adalimumab or infliximab. All received an induction regimen followed by maintenance therapy with infliximab 5 mg/kg every 8 weeks or adalimumab 40 mg every other week and provided at week 0 and 14 fecal and blood samples for determination serum CRP, serum and fecal calprotectin and S100A12 levels. RESULTS: Clinical remission at week 14 (responders) was achieved in 21 patients and among them, 12 were still in steroid-free clinical remission at week 52. Median serum S100A12 and fCal concentrations significantly drop only in responders from week 0 to week 14 after induction, whereas serum calprotectin and fecal S100A12 levels failed to differ significantly. Fecal calprotectin levels at week 14 had the highest discriminant validity to predict clinical remission within 1 year after induction (area under the curve = 0.87) followed by fecal, serum S100A12, and serum calprotectin (area under the ROC curve = 0.70, 0.70, and 0.68, respectively). A cutoff of 82 mug/g for fCal at week 14 had a sensitivity and specificity of 93% and 75%, respectively, to predict clinical remission within 1 year of therapy. CONCLUSIONS: Serum S100A12 level and fCal are reliable markers associated with response to induction therapy with anti-TNF. Fecal calprotectin was the best for predicting clinical remission in CD under maintenance therapy

ROS Production and Distribution: A New Paradigm to Explain the Differential Effects of X-ray and Carbon Ion Irradiation on Cancer Stem Cell Migration and Invasion

Although conventional radiotherapy promotes the migration/invasion of cancer stem cells (CSCs) under normoxia, carbon ion (C-ion) irradiation actually decreases these processes. Unraveling the mechanisms of this discrepancy, particularly under the hypoxic conditions that pertain in niches where CSCs are preferentially localized, would provide a better understanding of the origins of metastases. Invasion/migration, proteins involved in epithelial-to-mesenchymal transition (EMT), and expression of MMP-2 and HIF-1α were quantified in the CSC subpopulations of two head-and-neck squamous cell carcinoma (HNSCC) cell lines irradiated with X-rays or C-ions. X-rays triggered HNSCC-CSC migration/invasion under normoxia, however this effect was significantly attenuated under hypoxia. C-ions induced fewer of these processes in both oxygenation conditions. The differential response to C-ions was associated with a lack of HIF-1α stabilization, MMP-2 expression, or activation of kinases of the main EMT signaling pathways. Furthermore, we demonstrated a major role of reactive oxygen species (ROS) in the triggering of invasion/migration in response to X-rays. Monte-Carlo simulations demonstrated that HO● radicals are quantitatively higher after C-ions than after X-rays, however they are very differently distributed within cells. We postulate that the uniform distribution of ROS after X-rays induces the mechanisms leading to invasion/migration, which ROS concentrated in C-ion tracks are unable to trigger

Delta Hemolysin and Phenol-Soluble Modulins, but Not Alpha Hemolysin or Panton-Valentine Leukocidin, Induce Mast Cell Activation

International audienceMast cells are located at host interfaces, such as the skin, and contribute to the first-line defense against pathogens by releasing soluble mediators, including those that induce itching and scratching behavior. Here, we show that delta-hemolysin (Hld) and phenol soluble modulins (PSMs) PSMα1 and PSMα3, but not alpha-hemolysin (Hla) or Panton-Valentine leukocidin (PVL), induce dose-dependent tryptase, and lactate dehydrogenase (LDH) release by the HMC-1 human mast cell line. Using supernatants from isogenic strains, we verified that tryptase and LDH release was Hld- and PSMα-dependent. PSMα1 and Hld production was detected in 65 and 17% of human Staphylococcus aureus-infected skin abscess specimens, respectively, but they were produced in vitro by all clinical isolates. The results suggest that Hld and PSM-α1 produced in vivo during S. aureus skin infections induce the release of mast cell mediators responsible for itching and scratching behavior, which may enhance skin to skin transmission of S. aureus via the hands. As Hld and PSMs are upregulated by accessory gene regulator (agr), their association may contribute to the elective transmission of S. aureus strains with a functional agr system

ROS Production and Distribution: A New Paradigm to Explain the Differential Effects of X-ray and Carbon Ion Irradiation on Cancer Stem Cell Migration and Invasion

Although conventional radiotherapy promotes the migration/invasion of cancer stem cells (CSCs) under normoxia, carbon ion (C-ion) irradiation actually decreases these processes. Unraveling the mechanisms of this discrepancy, particularly under the hypoxic conditions that pertain in niches where CSCs are preferentially localized, would provide a better understanding of the origins of metastases. Invasion/migration, proteins involved in epithelial-to-mesenchymal transition (EMT), and expression of MMP-2 and HIF-1α were quantified in the CSC subpopulations of two head-and-neck squamous cell carcinoma (HNSCC) cell lines irradiated with X-rays or C-ions. X-rays triggered HNSCC-CSC migration/invasion under normoxia, however this effect was significantly attenuated under hypoxia. C-ions induced fewer of these processes in both oxygenation conditions. The differential response to C-ions was associated with a lack of HIF-1α stabilization, MMP-2 expression, or activation of kinases of the main EMT signaling pathways. Furthermore, we demonstrated a major role of reactive oxygen species (ROS) in the triggering of invasion/migration in response to X-rays. Monte-Carlo simulations demonstrated that HO・ radicals are quantitatively higher after C-ions than after X-rays, however they are very differently distributed within cells. We postulate that the uniform distribution of ROS after X-rays induces the mechanisms leading to invasion/migration, which ROS concentrated in C-ion tracks are unable to trigger

Milk polar lipids favorably alter circulating and intestinal ceramide and sphingomyelin species in postmenopausal women

International audienceBACKGROUND. High circulating levels of ceramides (Cer) and sphingomyelins (SM) have been associated with cardiometabolic diseases. The consumption of whole-fat dairy products, which naturally contain such polar lipids (PL), is associated with health benefits, but the impact on sphingolipidome remains unknown. We investigated how milk PL supplementation impacts circulating and intestinal SM and Cer composition in association with improvement of cardiovascular markers.METHODS. In a 4 week-randomized double-blind controlled study, 58 postmenopausal women consumed daily a cream cheese containing 0, 3 or 5 g of milk PL. Postprandial metabolic explorations were performed before and after the supplementation. SM and Cer species were analyzed in serum, intestine-derived chylomicrons and feces. The ileal content of 4 ileostomy patients was also explored after milk PL intake in a crossover double-blind study.RESULTS. Milk PL consumption decreased serum atherogenic C24:1 Cer (Pgroup = 0.033), C16:1 (Pgroup = 0.007) and C18:1 (Pgroup = 0.003) SM species. Changes in serum C16+18 SM species were positively correlated with the reduction of total cholesterol (r = 0.706, P < 0.001), LDL-C (r = 0.666, P < 0.001) and ApoB (r = 0.705, P < 0.001). Milk PL decreased the concentration in chylomicrons of total SM (Pgroup < 0.0001) and of C24:1 Cer (Pgroup = 0.001). Saturated SM and Cer species, which are also the major species found in milk PL-enriched cheeses, increased in ileal efflux and feces. There was a marked increase in total fecal Cer after milk PL supplementation (Pgroup = 0.0002). Milk PL also modulated the abundance of some specific SM and Cer species in ileal efflux and feces, suggesting differential absorption and metabolization processes in the gut.CONCLUSION. These data demonstrate that milk PL supplementation decreases atherogenic SM and Cer species associated with an improvement of cardiovascular risk markers. Our findings bring new insights on sphingolipid metabolism in the gastrointestinal tract, especially Cer as such signaling molecules potentially participating in the beneficial effect of milk PL. ClinicalTrials.gov, NCT02099032, NCT02146339