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Alcohol-related problems are typically associated with medical, economic and social issues. The reduction of the problem can only occur if effective strategies to prevent, diagnose and treat alcohol abuse are developed. Real help could come from laboratory tests that represent objective evidence of alcohol problems. Authors present new biomarkers as Carbohydrate-deficient transferrin (CDT), Ethyl Glucuronide and Ethyl Sulfate ( EtG and EtS)), Fatty Acids Ethyl Esters (FAEE) anf Phosphatidy ethanol (PEth) in differents biological samples.

Premesse L’utilizzo di sistemi di cromatografia liquida ultra veloce, non accoppiata a un rivelatore di massa, consentirebbe il dosaggio accurato delle differenti forme di vitamina D, proprio dei metodi cromatografici, in tempi rapidi, con produttivita simile a quella dei metodi immunometrici, senza richiedere investimenti iniziali elevati generalmente tipici dei sistemi LC/MS/MS. In questo studio, utilizzando plasma e siero come matrici, abbiamo confrontato un nuovo metodo cromatografico liquido ultraveloce (UPLC) con un metodo immunometrico e uno cromatografico tradizionale (HPLC).

Aims: Contrasting data are available on the diagnostic accuracy of carbohydrate-deficient transferrin (CDT) during pregnancy. These differences may depend in part on how CDT was evaluated and expressed. Here, we report on variations of CDT levels in pregnant women using the high performance liquid chromatography (HPLC) candidate reference method. Methods: Alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, mean corpuscular volume, serum transferrin, urine and serum ethyl glucuronide and CDT were measured in 64 women, self-reporting as non-alcohol abusers (age: median 34, IQR: 28–38), at different stages of normal pregnancy (gestational weeks: median 28, IQR: 8–33). CDT was expressed as percentage of disialotransferrin to total transferrin (%CDT). Results: Transferrin was associated with both %CDT ( r = 0.66; P

AIMS: Carbohydrate-deficient transferrin (CDT) has become widely used in traffic medicine to detect chronic alcohol abuse among subjects applying for driving-license renewal or regranting. By defining cut-off values in a large population of abstainers and moderate drinkers, we report on CDT, GGT-CDT (a combination of gamma-glutamylaminotransferase (GGT) and CDT) and the association between blood alcohol concentration (BAC) and CDT among Italian drivers. METHODS: CDT was evaluated by a high performance liquid chromatography (HPLC)-based commercial kit in 652 abstainers or moderate drinkers, 603 drivers applying for driving-license regranting after a rehabilitation programme and 105 drivers involved in car accidents with blood alcohol concentration higher than the legal limit used in Italy (BAC >0.5g/l). GGT-CDT was calculated according to Sillanaukee and Olsson and Niemela. BAC has been assessed by gas chromatography-mass spectrometry. RESULTS: A common CDT cut-off (1.8%) and gender-specific GGT-CDT cut-off (4.15% for males, 3.56% for females) were calculated as 99.9th percentiles of the control population. Also, 3% and 27% of subjects were classified as CDT positive respectively among drivers applying for license regranting and drivers involved in car accidents. A significant association between BAC and both CDT values and CDT positivity was found, with a frequency up to 49% of CDT samples, suggesting chronic alcohol abuse, among drivers with BAC >2.5g/l. Concordance between CDT and GGT-CDT was only moderate (kappa = 0.44), with CDT performing better than GGT-CDT. CONCLUSIONS: A relevant proportion of drivers with high BAC are chronic abusers. GGT-CDT, previously validated with CDT immunoassays, should not be applied to traffic medicine in its current form and its performances re-evaluated with CDT measured by HPLC. Language: en

To investigate the role of suPAR in patients with sepsis admitted to the Emergency Department (ED). We performed multicentre prospective trial including patients admitted to the ED of three different Italian hospitals. Patients were studied upon admission on day 1, 2, 4 and 7. They were subdivided into two groups: sepsis (group 1) and severe sepsis or septic shock (group 2). The two groups were comparable for age, gender and CRP level on day 1. Patients with severe sepsis or septic shock displayed significantly higher baseline levels of suPAR, PCT and lactate. In both groups, suPAR decreased across the time (p < 0.0005). Group 1 was not different from group 2 (p = 0.545) in mortality at 7 days, while group 2 had higher mortality at 30 days than group 1 (p = 0.022). At the multivariate analysis, lactate1 (p = 0.012) and age (p = 0.019) were independent predictors of mortality at 7 days, whereas suPAR1 (p = 0.023) and age (p = 0.032) were independent predictors of mortality at 30 days. Lactate and suPAR resulted the most predictive biomarkers in the risk stratification of patients with suspected infection initially admitted to the ED, according to their role in predicting 7- and 30-day mortality, respectively.

Background Therapeutic drug monitoring of the anticonvulsant levetiracetam may be indicated in patients with conditions that may alter pharmacokinetic characteristics, for tailoring individual dosage regimens or to investigate patient compliance. In this study, the Bio-Rad high-performance liquid chromatography (HPLC) method (in-use method) and the ARK immunoassay method (new method) for levetiracetam monitoring in serum were compared. Methods Levetiracetam concentrations were determined in 63 samples using: (1) "Levetiracetam by HPLC" kit by Bio-Rad (Hercules, CA) on the Agilent 1100 HPLC system, and (2) "ARK Levetiracetam" immunoassay by ARK Diagnostics Inc (Fremont, CA) on the CDx90 platform by ThermoFisher Scientific Inc. Results Within-laboratory imprecision and bias of the new method evaluated over a 20-day period were 7.4% and 0.5% at 7.5 mcg/mL, 4.5% and 1.9% at 30 mcg/mL, and 3.1% and 2.0% at 75 mcg/mL. Passing-Bablok regression analysis (X:Bio-Rad; Y:Ark) showed a nonsignificant intercept of 0.16 [95% confidence interval (CI), -0.55 to 0.72] and a slope marginally significantly different from unity of 0.95 (95% CI, 0.90-0.99), which suggested minimum proportional systematic error. In agreement, Bland-Altman analysis showed minimum systematic bias of 1.0 mcg/mL (95% CI, 0.32-1.69) with 95% of the HPLC-Ark differences ranging from -4.3 (95% CI, -5.52 to -3.16) to 6.3 (95% CI, 5.16-7.52). Our data showed that the 2 methods were identical both within inherent imprecision and analytical quality specifications (maximum allowable error 15%). Conclusions The new Ark method on the CDx platform is acceptable and may be used to measure serum levetiracetam concentrations routinely.

The evaluation of the age-specific distribution of transferrin glycoforms in paediatric patients may help in defining reference intervals which are critical for an improved and earlier diagnosis.Serum samples from 224 children (age: 2 months-14 years) were analyzed by HPLC (Bio-Rad CDT/HPLC kit) and glycoforms expressed as percentage of the total area of transferrin (Tf).Asialo- and Monosialo-Tf were not detectable in any patient. Medians (IQR) were respectively 0.92% (0.80-1.04%) for Disialo-Tf; 3.47% (2.69-4.18%) for Trisialo-Tf; 82.54% (81.32-83.53%) for Tetrasialo-Tf; 12.73% (11.91-14.09%) for Pentasialo-Tf. Statistically significant differences in Trisialo-Tf (p0.0005), Tetrasialo-Tf (p = 0.001), Pentasialo-Tf (p0.0005), but not in Disialo-Tf, were observed between the age groups.Age-specific Disialo-Tf cut-offs are not necessary. In children 1.3% and 6.4% may be suggested as upper limits of normal range to detect increases of Disialo- and Trisialo-Tf. The presence of Asialo- and Monosialo-Tf should be considered an abnormal finding and prompt further investigations.

Abstract 5094 Background. Leptomeningeal involvement is an adverse complication of haematological malignancies, acute leukemias and lymphomas. The diagnostic gold standard to identify cerebrospinal fluid (CSF) involvement is conventional cytological analysis, technique characterized by low sensitivity and specificity. Recent papers make evidence that the efficiency of CSF involvement detection could improve by use of Flow Cytometry (FCM) immunophenotypic analysis of CSF samples. We describe the FCM method used in our hospital and we compare the results with cytological findings. Methods. CSF samples should be processed immediately, within 1 hour after collection to avoid cell's deterioration; in B Non Hodglin Lymphomas (B-NHL) we use two steps FCM approach, in four color staining (FITC/PE/PerCP/APC). The first step was two tubes: kappa/lambda/CD45/CD19 and CD5/CD20/CD45/CD3 or CD3/CD20/CD45/CD10 on the basis of a previous know immunophenotype or not. If a pathological population is identified, we complete immunophenotype characterization with a second step, using antibody panels suitable for the suspected malignancy. In case of suspected meningeal relapse of Acute Leukemia we use a monoclonal antibody combination select for Minimal Residual Disease. Results. Between 2006 and 2009 we studied by FCM analysis 53 CSF samples of 36 patients. 7 CSF of 29 B-NHL newly diagnosed resulted positive at FCM analysis (24,1%), 1 CSF of 7 Acute Leukemias shows disease relapse. The conventional cytology was positive only in 1 case of B-NHL. Conclusion. Our study confirms that immunophenotypic flow cytometric analysis of CSF cells markedly improves sensitivity to identify CSF involvement in haematological malignancies. Disclosures: No relevant conflicts of interest to declare.