Firenze, Italy
Firenze, Italy

Time filter

Source Type

Poli G.,University of Florence | Guasti D.,University of Florence | Rapizzi E.,University of Florence | Fucci R.,University of Florence | And 7 more authors.
Endocrine-Related Cancer | Year: 2013

At present, mitotane (MTT) represents the first-line pharmacological approach for the treatment of advanced adrenocortical carcinoma (ACC). Despite clear evidence that the drug can reduce the clinical signs of steroid excess in secreting ACC, the mechanism mediating the possible toxic effect of MTT on tumor cells still remains obscure. This study investigated the intracellular events underlying the toxic effect of MTT by studying qualitative and quantitative alterations in mitochondrial morphology and functions in human adrenocortical cancer cell lines, H295R and SW13. Increasing concentrations of MTT resulted in rapid intracellular accumulation and conversion of the drug. Cytostatic and cytotoxic effects were evident at doses corresponding to the therapeutic window (30-50 μM) through an apoptotic mechanism involving caspase 3/7. Electron microscopic analysis of cell mitochondria displayed MTT-induced dose- and time-dependent alterations in the morphology of the organelle. These alterations were characterized by a marked swelling and a decrease in the number of respiratory cristae, accompanied by a significant depolarization of the mitochondrial membrane potential, finally leading to the disruption of the organelle. A drastic reduction of oxygen consumption was observed due to mitochondrial membrane damage, which was accompanied by a decrease in the levels of VDAC1 integral membrane channel. These findings contribute to better understand the intracellular mechanism of action of MTT in ACC cells, showing that its cytotoxic effect seems to be mainly mediated by an apoptotic process activated by the disruption of mitochondria. © 2013 Society for Endocrinology.


Claudia C.,Azienda Ospedaliero | Claudia R.,Azienda Ospedaliero | Agostino O.,General Laboratory of Clinical Biochemistry | Agostino O.,Laboratorio Generale | And 2 more authors.
Journal of Trauma - Injury, Infection and Critical Care | Year: 2011

Background: Head injury represents one of the most important and frequent traumatic pathology in the emergency department. Among the different risk factors, preinjury use of warfarin has received considerable attention in trauma literature. The aim of this study was to identify further risk indicators of intracranial hemorrhage (ICH) to improve risk stratification of warfarinized patients with minor head injuries. Methods: Medical records of 1,554 adult patients with minor head injuries evaluated by the Emergency Department of Azienda Ospedaliera, Universitaria Careggi from January 2007 to February 2008 were analyzed retrospectively. All the patients included in the study were subjected to blood tests. The international normalized ratio (INR) measured on admission was correlated with the results of head computed tomography scan. Results: Of the 1,410 patients included in the study, 75 (5.2%) were warfarin anticoagulated at the time of trauma. The INR measured on admission was 2.37 ± 1.04 (mean ± standard deviation), and this value was significantly associated with occurrence of ICH after head trauma (r = 0.37; p < 0.005). For 12 (of 75) patients of this group, the findings of the computed tomography scans were positive. The receiver operating characteristic curve show that the most effective INR cutoff value was 2.43, with a sensitivity of 92%, a specificity of 66%, and positive and negative predictive values of 33% and 97%, respectively. Conclusion: This study highlights the strong relationship between INR values and the probability of ICH, as shown in previous studies. The high negative predictive value of the identified cutoff, if confirmed, could be used to exclude ICH. Copyright © 2011 by Lippincott Williams & Wilkins.


Graziani M.S.,Azienda Ospedaliera Universitaria Integrata di Verona | Secchiero S.,University of Padua | Terreni A.,Laboratorio Generale | Caldini A.,Laboratorio Generale | Panteghini M.,University of Milan
Biochimica Clinica | Year: 2015

The diagnosis and classification of CKD are based on laboratory tests. Aim of this paper is to examine different aspects of the laboratory contribution to verify their harmonization at national level. We review relationships between laboratory and clinical organizations, the role of 2012 Kidney Disease Improving Global Outcomes (KDIGO) guidelines, the quality of creatinine and urine albumin measurements, the status of estimated glomerular filtration rate (eGFR) reporting, the use of cystatin C and testing plans. Questionnaires examining different aspects of the CKD diagnostics were sent out and EQAS for creatinine and urine albumin measurements were carried out. For creatinine measurement, enzymatic assays show the best performance, while for urine albumin a bias still exists between different methods. The eGFR is routinely reported by 75% of surveyed laboratories, but only 15% of them use the equation derived by the CKDEpidemiology Collaboration (CKD-EPI) study. For urine albumin, the recommended first morning void sample is used by ∼60% of laboratories, but the wrong terminology of "microalbuminuria" is still used by >40% of them. Cystatin C is offered by a minority of laboratories. In conclusion, even if an improvement can be observed during the recent years, efforts for a better alignment to international recommendations are needed. Often they just require cultural and organizational changes, without the availability of additional economic resources.


Terreni A.,Laboratorio Generale | Caldini A.,Laboratorio Generale | Graziani M.S.,Azienda Ospedaliera Universitaria Integrata di Verona | Merlini G.,University of Pavia
Biochimica Clinica | Year: 2015

Protein diagnostics is central in the management of subjects with monoclonal gammopathy. Laboratory should provide the most useful information to ensure the best patient outcome. To assess if recommendations issued after the 2007 survey have impacted on Italian laboratories contributing to a better harmonization of the post-analytical phase, the SIBioC Proteins Study Group has repeated a similar survey in February 2015. Twenty questions were electronically submitted to all SIBioC members using the software "Survey monkey". 103 responses were collected, corresponding to ∼6% of Italian laboratories. 47% of laboratories add an appropriate comment to the serum protein electrophoresis report when no monoclonal component (MC) is detected (36% in 2007). MC are correctly defined by 63% of the laboratories; however, 11% reports MC as "thickening" or "asymmetry" or "homogeneous peak". These ambiguous terms were used by roughly the same percentage (14%) in 2007. In 2015, the number of laboratories performing a MC typing only when requested by the clinician is reduced by 10% when compared to 2007. In both surveys, the percentage of laboratories performing and reporting the MC quantification is 77%. The worse results were obtained for Bence Jones protein (BJP) determination (not investigated in 2007): only 66% of laboratories utilize the immunofixation to detect the BJP and 57% do not quantify the protein. Although some progresses in harmonization of reporting are observed in CM testing over years, there is still room for significant improvement.


Rapi S.,Laboratorio Generale | Fraser C.G.,University of Dundee | Cellai F.,Instituto Studio Prevenzione Oncologica | Berardi M.,Laboratorio Generale | Rubeca T.,Instituto Studio Prevenzione Oncologica
Biochimica Clinica | Year: 2015

Sampling of feces is strongly affected by the lack of harmonisation, with differences up to 20 times in the mass collected for immunological tests for fecal hemoglobin (Hb) used in colorectal cancer screening. Aim of this study was to acquire information on fecal sampling and on the interaction between feces and analytical methods to obtain a reference design for a sampling dipstick. Bias and imprecision of sample collection dipsticks were estimated using gravimetry. Dissolution times of feces were monitored throughout the study. The effect of increasing amount of feces on Hb concentrations was investigated in saline and buffers of different manufacturers using a single analytical method (OC-Sensor, Eiken Chemical Co.). Fecal mass recovered with different devices ranged from 56 to 121% of declared amount (CV range: 8.6/31.1%). Time of dissolution up to 2 h was observed when lumps of materials were collected. In saline a rapid decrease of Hb values was observed, which was related to the overall amount of feces. Increased Hb values were observed by adding feces to manufacturers' buffers. Solubilisation time, bias and imprecision of sampling of feces were related to device design. Analytical methods are designed to use specific ratios between feces and buffers. The introduction of a standard dipstick design to reduce preanalytical variability may represent a crucial step for fecal test harmonization.


Pezzati P.,Laboratorio Generale | Balboni F.,Instituto Fiorentino Of Cura E Assistenza
Biochimica Clinica | Year: 2015

Copeptin (CP) is a 39 amino acid glycopeptide including the C-terminal domain of the arginine-vasopressine precursor. CP is known to be a stable and sensitive surrogate marker for arginine-vasopressine release and it has been investigated as possible diagnostic and prognostic marker in cardiovascular and cerebrovascular diseases. In particular, the role of CP in the early diagnosis of acute myocardial infarction has been widely investigated and various systematic reviews are currently available. CP associated with cardiac troponin T has been reported to carry a good diagnostic accuracy. These data need, however, to be revised according to the introduction of highly sensitive assays for measuring troponins. Although some evidence on the prognostic value of CP in various cardiovascular frameworks is available, natriuretic peptides maintain a leading role. The research on CP is currently investigating its prognostic value in stroke and transient ischemic attack. However, due to the small sample size of studies and the lack of sound evidence, there are no minimal indications for requesting CP in this framework.


The IMWG has recently updated the disease definition of multiple myeloma, by adding validated biomarkers to the existing requirements of organ damage (hypercalcemia, renal insufficiency, anemia, bone lesions). These changes are based on the identification of biomarkers able to detect the subset of patients with smouldering multiple myeloma at imminent risk of developing organ damage and should, therefore, be considered for therapy. Considering that the clinical laboratory is involved in the measurement of these new markers, this paper is aimed to illustrate the proposed changes giving at the same time some indications for their accurate measurements. As for the organ damage, the major change is related to the evaluation of renal function: the new criteria include the estimation of the glomerular filtration rate using established formulas (eGFR) rather than the use of serum creatinine concentrations alone, as previously indicated. The diagnosis of renal insufficiency requires an eGFR <40 mL/min/1.73 m2. The criteria for anemia and hypercalcemia remain unchanged. As biomarker of malignancy, a ratio >100 of involved to uninvolved serum free light chains is recognized. Another relevant modification is the elimination of the monoclonal protein quantification; it is based on the consideration that an important percentage of patients with multiple myeloma does not show a serum or urine monoclonal protein. Other changes based on imaging techniques or bone marrow examination do not involve the clinical laboratory and are not discussed in this paper. Additional biomarkers will probably be indentified in the near future, but they need to be validated by more independent studies. © 2015 Biochimica Clinica.


Mussap M.,Instituto Nazionale Per la Ricerca Sul Cancro | Graziani M.S.,Azienda Ospedaliera Universitaria Integrata di Verona | Caldini A.,Laboratorio Generale | Dolci A.,UOC Patologia Clinica | Merlini G.,University of Pavia
Biochimica Clinica | Year: 2014

The contrast media, widely used in imaging diagnostics, show a favorable safety profile. As the presence of pre-existing disease is considered a risk factor for adverse events, patients should be carefully evaluated prior to the procedure. The aim of this consensus document is to recommend appropriate biochemical tests to be performed for an early recognition of individuals at higher risk of contrast media nephrotoxicity. This condition is defined by an increase of serum creatinine concentrations of at least 0.50 mg/dL and/or 25% within 3-4 days from contrast media exposure. The most important risk factor is renal insufficiency [estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m2 or serum creatinine >1.50 mg/dL]. Other risk factors are age >75 years, dehydration, diabetes, heart failure and anemia. Monoclonal gammopathies, multiple myeloma, Waldenström macroglobulinemia and amyloidosis are not considered risk factors per se. On the basis of available guidelines, it is recommended: a) prior to the examination, to measure serum creatinine baseline with a method traceable to the international reference measurement system and report its concentration together with the eGFR using the Chronic Kidney Disease - Epidemiology Collaboration (CKD-EPI) equation; b) for monitoring, to measure serum creatinine more than once calculating the delta from the baseline value: if serum creatinine increases >5%, repeat the test within 48- 72 h. Performing of laboratory tests to exclude the presence of monoclonal gammopathies (i.e., serum protein electrophoresis, Bence Jones protein determination, serum free light chain measurements) is not required.


We present here a recommendation for the urine albumin measurement in diabetic nephropathy. The suggestions are derived from those by the IFCC-National Kidney Disease Education Programme (NKDEP) Working Group on Standardization of Urine Albumin Assays (WG-SAU) and are based on the best available evidence. They cover preanalytical, analytical, and post-analytical phases.


PubMed | Laboratorio Generale
Type: Journal Article | Journal: Clinical biochemistry | Year: 2016

The recent guideline for the evaluation and management of Chronic Kidney Disease recommends assessing GFR employing equations based on serum creatinine; despite this, creatinine clearance 24-hour urine collection is used routinely in many settings. In this study we compared the classification assessed from CrCl (creatinine clearance 24h urine collection) and e-GFR calculated with CKD-EPI or MDRD formulas.In this retrospective study we analyze consecutive laboratory data: creatinine clearance 24h urine collection, serum creatinine and demographic data such as sex and age from 15,777 patients >18 years of age collected from 2011 to 2013 in our laboratory at Careggi Hospital. The results were then compared to the estimated GFR calculated with the equations according to the recent treatment guidelines. Consecutive and retrospective laboratory data (creatinine clearance 24h urine collection, serum creatinine and, demographic data such as sex and age) from 15,777 patients >18 years of age seen at Careggi Hospital were collected.Comparison between e-GFR calculated with CKD-EPI or MDRD formulas and GFR according CrCl determinations and bias [95% CI] were 11.34 [-47,4/70.1] and 11.4 [-50.2/73] respectively. The concordance for 18/65 years aged group when compared with e-GFR classification between MDRD vs CKDEPI, MDRD vs CrCl and CKD-EPI vs CrCl were 0.78, 0.34, and 0.41 respectively, while in the 65/110years aged group the concordance Kappas were 0.84, 0.38, and 0.36 respectively.The use of CrCl provides a different classification than the estimation of GFR using a prediction equation. The CrCl is unreliable when it is necessary to identify CKD subjects with decrease of GFR of 5ml/min/1.73m(2)/year.

Loading Laboratorio Generale collaborators
Loading Laboratorio Generale collaborators