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San Raffaele Cimena, Italy

Bernardi G.,Laboratorio Of Patologia Clinica E Genetica Medica | Passerini G.,Diagnostica e Ricerca San Raffaele | Previtali G.,Presidio | Ciusani E.,Laboratorio Of Patologia Clinica E Genetica Medica
Biochimica Clinica | Year: 2014

Pathological CSF leakage outside central nervous system is a very dangerous situation with high risk of meningitis and cerebral abscess. Rhinorrhoea and post-surgical CSF leakage are the most frequent conditions. Diagnosis is made by combination of imaging procedures, radionucleide cisternography via lumbar puncture (fluorescein) and biochemical tests, which utilize markers suggesting the presence of CSF in suspected fluid. Low CSF glucose concentrations suggest rhinorrhoea, as glucose is absent in nose secretion, but cannot be used in post-surgical leakage, as blood is nearly almost present. CSF proteins are the best biomarkers and intrathecal synthesized β-trace protein and transferrin are the best choice. For quantitative analysis, nephelometric β-trace protein measurement has the best performance, as it can be easily automatically performed, also for stat analysis, but relatively high volume of sample is needed. Isoelectricfocusing or high resolution electrophoresis followed by immunodetection are the most sensitive and specific methods for detecting asialotransferrin, but they are time consuming, unsuitable for stat analysis, even if they need low amounts of sample. Other quantitative tests include prealbumin/albumin ratio, having insufficient sensitivity in blood contaminated samples, and zone electrophoresis of protein pattern that, however, has too low sensitivity. New methods like capillary electrophoresis have been recently proposed.

Bernardi G.,Laboratorio Of Patologia Clinica E Genetica Medica | Brunati P.,Laboratorio Analisi Chimico Cliniche e Microbiologia | Biagioli T.,Laboratorio Generale | Buoro S.,Laboratorio Analisi Chimico Cliniche | And 8 more authors.
Biochimica Clinica | Year: 2014

The laboratory investigation of CSF has been developed over the years as a diagnostic tool for many neurological diseases. Although minimally invasive, CSF is obtained with a traumatic procedure; therefore, the whole laboratory process should be established to maximize the analytical performance. Based on the review of international guidelines and on the experience developed by members of the SIBioC Working Group, the present document provides practical information for laboratory professionals to better address the CSF analysis in different diagnostic situations. The report faces the pathophysiologic meaning of the determination of biochemical parameters, such as glucose, lactate, albumin, immunoglobulins, β-amyloid, tau protein, and the cellular content, providing also evidence on the proper methodological approach. Quantitative and qualitative CSF parameters useful to diagnose an inflammatory process of the central nervous system are discussed, particularly with reference to multiple sclerosis. Indications on how laboratory data should be presented to meet international recommendations are also included.

Seminari E.,Clinica di Malattie Infettive e Tropicali | de Silvestri A.,Fondazione IRCCS Policlinico San Matteo | Meini G.,University of Siena | Callegaro A.,Microbiologia e Virologia | And 5 more authors.
Current HIV Research | Year: 2012

Objective: The aim of the present study was to evaluate the virological response to a new antiretroviral treatment (ART2) after failure of a nonnucleoside reverse transcriptase inhibitor (NNRTI) plus two nucleoside reverse transcriptase inhibitors (NRTIs)-containing regimen. Design: Retrospective observational study based on the Italian ARCA cohort database. Adult patients were included if they had a virological failure (defined as plasma viral load above 500 copies/ml in two subsequent visits) while on a treatment with one NNRTI plus 2 NRTIs, had an available HIV genotype. Results: Patients on ART2 were followed up for 791 person/year and median follow up was 10.8 months(IQR 5.2-26). Variables associated with reduced risk of ART2 virological failure at univariable analysis had started the treatment in recent years (HR 0.90; 95% CI 0.86-0.94, p<0.0001) and duration of previous NNRTI treatment (HR 0.995; 95%CI0 990-0.990, p=0.045). Variables associated with increased risk of virological failure of ART2 were a higher plasma viral load (pVL) at baseline(HR 1.2; 95% CI 1.07-1.34, p=0.002) and the type of treatment, in particular an unboosted PIcontaining regimen vs. a boosted PI-containing regimen(HR 1.6; 95%CI 1.25-2.04 p<0.0001) and a non-PI-containing vs. a boosted PI-containing regimen (HR 1.56; 95% CI 1.25-1.96, p<0.0001). At multivariable analysis, year of ART2 start, pVL at NNRTI failure as well as using a boosted PI remained statistically significant predictors. Conclusion: This study highlights the role of drugs with high genetic barrier, such as boosted PI as a cornerstone to build a new antiretroviral treatment in patients failing a NNRTI based regimen. © 2012 Bentham Science Publishers.

Lombardi G.,Laboratory of Experimental Biochemistry and Molecular Biology | Lanteri P.,Laboratory of Experimental Biochemistry and Molecular Biology | Fiorella P.L.,Commissione Tutela della Salute Federazione Ciclistica Italiana | Simonetto L.,Commissione Tutela della Salute Federazione Ciclistica Italiana | And 5 more authors.
PLoS ONE | Year: 2013

Cycling stage races are strenuous endurance events during which exercise-induced variations in hematological parameters are consistently observed. However, specific literature on such changes is scarce and published data have been derived from small samples of athletes. The aims of this study were: (1) to determine the hematological response to middle-term strenuous endurance; and (2) to determine whether a relationship exists between the athlete-specific hematological profile and final placement in a cycling stage race. The study population was male professional cyclists (n = 253) competing in the 2010 (n = 144) and 2012 (n = 109) GiroBio 10-day stage races. Blood draws taken before the start of the race, at mid-race, and at end-race were performed in strict compliance with academic and anti-doping pre-analytical warnings. Blood chemistry included white blood cell, red blood cell, hemoglobin concentration, hematocrit, mean corpuscular volume (MCV), mean hemoglobin content (MCH), mean corpuscular hemoglobin content (MCHC), platelets, and reticulocyte relative and absolute counts. Compared to baseline values, erythrocyte, hemoglobin, hematocrit, MCHC, platelet and reticulocyte counts were all consistently lower at mid-race, but returned to normal by race-end, while leukocytes were increased in the final phase. MCV increased during both events. MCH increased in the first part to then return to baseline in the 2012 race. The calculated OFF-score consistently decreased in the first half of the race before increasing, but remained lower than the baseline value. The trends of variation in hematological parameters were substantially similar in both events. There was an inverse, albeit weak, relationship between placement and erythrocyte, platelet, hemoglobin, hematocrit and OFF-score values in the 2010, but not in the 2012 race. In conclusion, the data confirm that, in this large series of elite road cyclists, the strenuous effort a rider sustains during a stage race induces appreciable changes in the hematological profile. © 2013 Lombardi et al.

Carobene A.,Diagnostica e Ricerca San Raffaele | Ceriotti F.,Diagnostica e Ricerca San Raffaele | Infusino I.,University of Milan | Frusciante E.,University of Milan | Panteghini M.,University of Milan
Biochimica Clinica | Year: 2012

Creatinine determination in serum is a key indicator of kidney glomerular function. A reference measurement system for standardization of creatinine measurements is now available and virtually all IVD manufacturers have aligned their creatinine assays to this system. The aim of this work was to verify if and how these standardization efforts have improved the state of the art of creatinine determination in Italy through the analysis of Prolarit EQAS results using control materials with target values assigned by a traceable method (enzymatic assay calibrated against the NIST SRM 967). Results obtained during 2006, 2010, and 2011 schemes by participating laboratories showed a general good alignment at creatinine concentrations ∼2.00 mg/dL, with 2011 results - except for one method group - well inside the desirable bias (±4%). At higher concentrations, whereas the overall bias was small in 2010, for some groups using alkaline picrate (AP) methods it became significantly negative in 2011. The performance markedly worsens at creatinine physiologic concentrations, where a significant positive bias (up to-20%) is still present for most of the AP-based analytical systems. Unexpectedly, with few exceptions, no evident improvement in individual assay bias was noted from pre- (2006) to post-standardization (2011) periods. The enzymatic method groups were the only always presenting an acceptable bias for all concentration levels, in addition to showing the lowest between-laboratory variability. The number of laboratories using enzymatic methods, however, still remains only 7% of the total. In conclusion, our EQAS performance data indicate that most of the current "standardized" creatinine methods based on AP reaction do not perform well, mainly at the lower creatinine concentrations. This inaccuracy of creatinine measurements can adversely impact the estimation of glomerular filtration rate by equations and the evaluation of kidney function in pediatrics.

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