False negativity to carbohydrate-deficient transferrin and drugs: a clinical case

Introduction: In this work we report on the possible effect of the medical therapy on CDT concentration in a chronic alcohol abuser, with known medical history (July 2007 – April 2012) and alcohol abuse confirmed by relatives. Case history: At the end of 2007, patient displayed the following laboratory results: AST 137 U/L, ALT 120 U/L, GGT 434 U/L, MCV 101 fL and CDT 3.3%. On December 2007, after double coronary artery bypass surgery, he began a pharmacological treatment with amlodipine, perindopril, atorvastatin, isosorbide mononitrate, carvedilol, ticlopidine and pantoprazole. In the next months, until may 2011, the patient resumed alcohol abuse, as confirmed by relatives; however, CDT values were repeatedly found negative (0.8% and 1.1%) despite elevated transaminases and GGT, concurrent elevated ethyl glucuronide concentration (> 50 mg/L) and blood alcohol concentration (> 1 g/L). Alcohol consumption still continued despite increasing disulfiram doses ordered by an Alcohol Rehab Center. On May 2011, the patient was transferred to a private medical center where he currently lives. Conclusions: This study suggests the possibility that a medical therapy including different drugs may hamper the identification of chronic alcohol abusers by CDT

Probiotics-addicted low-protein diet for microbiota modulation in patients with advanced chronic kidney disease (ProLowCKD): A protocol of placebo-controlled randomized trial

Abstract Microbiota is a term coined to describe the population of bacteria, viruses and fungi that inhabit in symbiosis within a living host. A connection between unbalanced microbiota and chronic kidney disease has been established. In these patients, high levels of urea reach the intestine promoting the overgrowth of bacterial species that are prone to generate uremic toxins. Due to the high morbidity and mortality of this condition, a large number of therapeutic approaches to reduce inflammation and microbial uremic toxins have been proposed, with controversial results. A low protein diet, with a protein intake of 0.6–0.8 g/kg of body weight, is a useful and historically pursued option with this regard. The aim of our study is to evaluate, among patients with advanced renal failure not on dialysis, the synergic beneficial effects of this diet and the selected probiotics Bifidobacterium longum (mix DLBL) and Lactobacillus reuteri LRE02 (DSM 23878)

The search for a myocardial ischemia marker. The case of albumin modified by ischemia

The search for a suitable and reliable marker of myocardial ischemia has led to the proposal of employing the cobalt binding capacity of total serum assuming that ischemic modifications of circulating albumin would markedly decrease its ability to bind cobalt. This test (IMA, Ischemia-Modified Albumin) can be easily implemented on most of the technological platforms employed for clinical chemistry assays. It is simple and rapid and suitable for most of the diagnostic purposes of emergency medicine. This assay has been successfully employed to detect myocardial ischemia in various clinical studies and, despite a low specificity for myocardial ischemia, its high sensitivity and negative predictive value have pointed out its possible role as a first-line assay in clinical setting of patients with chest pain admitted to the Emergency Department. The low specificity toward ischemic events occurring to myocardium is challenging its use in clinical conditions associated with tissue ischemia or whole body hypoxia. Few caveat, however, must be taken into account. They include the clear identification of the proteins responsible for cobalt binding, the normalization of the results for the serum concentration of the binding sites, the standardizations of the assays employed. Large randomized trials are also absolutely needed to undoubtedly identify the potential and the limit of this new and promising biomarker

Clinical Mass Spectrometry in Immunosuppressant Analysis: Toward a Full Automation?

The analysis of immunosuppressive drugs allows the physician to monitor, and eventually correct, immunosuppressive therapy. The panel of molecules under evaluation includes cyclosporine A (CsA), tacrolimus, sirolimus, and everolimus. Initially, assays were performed by immunometric methods, but in the past few years this methodology has been largely superseded by a more accurate and specific technique, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), which is now considered the “gold standard” for immunosuppressant analysis. Both LC-MS/MS and often also immunoassays require a preanalytical manual sample preparation, which involves time-consuming sequential operations whose traceability is often hampered and adds up to the probability of gross errors. The aim of this work was to compare an “open” LC-MS/MS with a fully automated system, consisting of LC instrumentation combined with a triple quadrupole MS, named Thermo ScientificTM CascadionTM SM Clinical Analyzer (Cascadion). Such automated systems suit the requirements of the reference method and are designed to completely eliminate all of the manual procedures. More than 2000 immunosuppressant samples were analyzed both with the open LC-MS/MS and with Cascadion. Statistics allowed the evaluation of linearity, intra- and inter-assay CV%, bias %, limit of detection and of quantitation, and Passing–Bablok and Bland–Altman plots. Results indicated a good correlation between the two methods. In both cases, methods confirmed their suitability for diagnostic settings. Cascadion could provide support when the presence of specialized personnel is lacking, and/or when great productivity and continuous workflow are required

Evaluation of the brain-to-plasma distribution of AH-7921 and semi-quantitative analysis of its main metabolites in rats

Introduction: AH-7921 is a synthetic opioid, possibly associated to non-fatal intoxications and even deaths. Several metabolites were found in blood and urines of AH-7921 users, as well as in in vitro studies. No data are however available as regards the pharmacokinetic profile, in particular in the brain, of AH-7921 and its metabolites. Methods: Rats were treated intraperitoneally with 10 mg/kg AH-7921. Brain and plasma metabolites were firstly identified by an high resolution mass spectrometry LTQ-Orbitrap XL. Determination of drug concentrations, and semi-quantitative analysis of the main metabolites, were obtained by HPLC-MS/MS, using a triple quadrupole with ESI positive ionization in MRM mode. Results: Cmax was reached 30 min after treatment, either in plasma (205 ng/mL on average) and brain (2842 ng/g), with a brain-to-plasma ratio of ~20. Plasma and brain levels then declined with a similar t1/2 (~ 3 hours). Different metabolites (N-desmethylated, N-didesmethylated, hydroxylated, N-desmethylated hydroxylated and N-didesmethylated hydroxylated) were detected in plasma and brain already 5 minutes after dose and at the following time-points. In particular, the first two reached, at longer time-points, brain levels comparable or even higher than those of the parent compound. Notably, the N-didesmethylated metabolite showed a very slow elimination from brain tissue. Conclusions: The new psychoactive drug AH-7921, as well as its main metabolites, readily reach the brain with brain-to-plasma ratios of ~15-20. The pharmacokinetic data obtained for the two main metabolites suggests the need of a further characterization of their pharmacological activities, as well as of similar studies after repeated treatments