Orbassano, Italy
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Ascari E.,Laboratorio Analisi
Biochimica Clinica | Year: 2013

These drugs have a considerable degree of safety and efficacy, but they may present certain drawbacks, as they require frequent laboratory monitoring (unfractionated heparins and anti-vitamin K) or intravenous or subcutaneous administration (low molecular weight heparins). Due to these reasons, several studies have been performed to identify new anticoagulant drugs with characteristics of safety, efficacy and possibility of oral administration and no need of laboratory monitoring. New direct oral anticoagulants (DOAs) have as direct target thrombin (factor IIa) or factor-Xa inhibition; their safety and efficacy have been investigated in several phase III studies. DOAs in most advanced stage of release are dabigatran, rivaroxaban and apixaban; the first drug is a direct thrombin inhibitor, the others are direct factor-Xa inhibitors. DOAs do not require routine laboratory monitoring due to their high dose-response predictability; there is, however, a necessity to measure their anticoagulant effect in some particular situations. In those situations, it is important for the laboratory to consider: 1) how DOAs can interfere in the measurement of basic and more specialistic coagulation parameters; 2) which tests are the most suitable to detect the presence of the new drug; 3) which tests are the most useful to measure the anticoagulant effect; 4) which tests are the most suitable to monitor the antagonizing effect of some drugs ("reversal") in case of DOA overdose.


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.


Banfi G.,University of Milan | Lombardi G.,University of Milan | Colombini A.,University of Milan | Lippi G.,Laboratorio Analisi
Sports Medicine | Year: 2010

Bone mass can be viewed as the net product of two counteracting metabolic processes, bone formation and bone resorption, which allow the skeleton to carry out its principal functions: mechanical support of the body, calcium dynamic deposition and haemopoiesis. Besides radiological methods, several blood and urinary molecules have been identified as markers of bone metabolic activity for estimating the rates and direction of the biological activities governing bone turnover. The advantages for the use of bone metabolism markers are that they are potentially less dangerous than radiological determinations, are more sensitive to changes in bone metabolism than radiological methods and are easily collected and analysed. The disadvantages are that they have high biological variability.Physical exercise is a known source of bone turnover and is recommended for preventing osteoporosis and bone metabolism problems. There are numerous experiments on bone metabolism markers after acute exercise, but not after long-term training and during or after a whole competition season. Moreover, few studies on bone metabolism markers have evaluated their performance in elite and top-level athletes, who have a higher bone turnover than sedentary individuals.Despite discrepant results among studies, most have shown that short exercise is insufficient for modifying serum concentrations of bone metabolism markers. Marker variations are more evident after several hours or days after exercise, bone formation markers are more sensitive than bone resorption markers, and stimulation of osteoblast andor osteoclast functions is exercise dependent but the response is not immediate. The response depends on the type of exercise; the markers seem to be less sensitive to resistance exercise and the intensity of exercise is not discriminate. Comparisons between trained subjects and untrained controls have demonstrated the influence of exercise on bone turnover. During training, carboxy-terminal collagen cross-links (CTx), a bone resorption marker, was shown to be less sensitive than amino-terminal cross-linking telopeptide of type I collagen (NTx) and urinary pyridinolines, which were sensitive to anaerobic exercise. Whereas, the bone formation markers, bone alkaline phosphatase (BAP) and osteocalcin (OC) changed after 1 month and 2 months of an exercise programme, respectively. After 2 months, while BAP normalized, it was found to be sensitive to aerobic exercise and OC was found to be sensitive to anaerobic exercise.After prolonged training and competition, bone formation markers are found to change in sedentary subjects enrolled in a physical activity programme. Professional athletes show changes in bone formation markers depending on programme intensity, whereas bone resorption appears to stabilize. Crucial for long-term training, are the characteristics of exercise (e.g. weight-bearing, impact). © 2010 Adis Data Information BV. All rights reserved.


Agodi A.,University of Catania | Barchitta M.,University of Catania | Valenti G.,University of Catania | Marzagalli R.,University of Catania | And 2 more authors.
European Journal of Clinical Nutrition | Year: 2011

Background/Objectives: Folate has been recognized to ensure reproductive health and there is a growing body of epidemiological evidence suggesting that the methylenetetrahydrofolate reductase (MTHFR) 677T allele and reduced dietary folate may increase the risk of cervical cancer. The main focus of our survey was to investigate the distribution of the MTHFR C677T polymorphism in relation to women's year of birth and to assess their folate intake and folic acid supplementation. Subjects/Methods: During a 6-months period, 307 healthy women of childbearing age in Catania, Italy, were enrolled in the cross-sectional study. Folate intake was estimated by a semiquantitative food frequency questionnaire and DNA extracted from blood samples for MTHFR C677T genotyping. Results: A TT genotype frequency of 20.5% with an increase in the prevalence of the TT genotype in the cohort of women born since 1959 was shown. The prevalence of inadequate folate intake was 51.5%, significantly higher in non-pregnant women (83.4%) than in pregnant ones (12.3%) with a decrease during the three trimesters of pregnancy (from 25.7 to 5.0%; P=0.013). The use of folic acid supplements improved during the three trimester of pregnancy (from 71.4 to 95.0%; P=0.001). Conclusions: Healthy young women may have higher folate needs due to increasing prevalence of the T allele and reduced folate intake compared with older groups. However, clinicians should be cautious when recommending supplements to women in late pregnancy due to the possible implications in the pregnancy outcome. © 2011 Macmillan Publishers Limited All rights reserved.


Infusino I.,University of Milan | Infusino I.,Laboratorio Analisi | Valente C.,University of Milan | Dolci A.,University of Milan | Panteghini M.,University of Milan
Analytical and Bioanalytical Chemistry | Year: 2010

The purpose of measurement standardization is to achieve closer comparability of results obtained using different commercial systems. Regarding serum protein immunoassays, a reference preparation (BCR-470) was released in 1993 and adopted by manufacturers across the world to value-assign their assay calibrators for routine methods to reduce method-dependent variation. Moving from nephelometric (Beckman Immage 800) to turbidimetric determination (Roche Cobas c 501) of seven serum proteins, we preliminarily checked the comparability of results between the two systems. The study was performed according to the CLSI EP9-A protocol on 30 fresh sera, tested on each system in duplicate, and subdivided on two different days, without recalibration and using manufacturers' control materials to validate the runs. Both manufacturers' package inserts provide statements that kit calibrators are traceable to BCR-470. Suggested reference intervals are also the same. Although a fairly good correlation was observed (r=0.955), the comparison of ceruloplasmin methods produced evidence of highly significant proportional (regression slope, 0.572) and constant bias (intercept, 0.05 g/L). Absolute and percentage mean differences were -0.11 g/L (95% confidence interval (CI) -0.13 to -0.10 g/L) and -39.1% (CI -43.1 to -35.2%), respectively. No other evaluated proteins showed similar problems. Lacking a ceruloplasmin reference method, it is impossible to demonstrate that one of the two assays produces true ceruloplasmin values. The problem is, however, that results coming from the two assays are clearly not comparable. This may be either due to a lack of commutability of the reference material with biological samples in the evaluated assays or to calibration problems by manufacturers in one of the stages of the calibration hierarchy. © 2009 Springer-Verlag.


Tozzoli R.,Laboratorio Of Patologia Clinica | Bonaguri C.,Laboratorio Diagnostica Ematochimica | Melegari A.,Laboratorio Of Tossicologia E Diagnostica Avanzata | Antico A.,Laboratorio Analisi | And 2 more authors.
Clinical Chemistry and Laboratory Medicine | Year: 2013

The methods for detecting and measuring autoantibodies have evolved markedly in recent years, encompassing three generations of analytical technologies. Many different immunoassay methods have been developed and used for research and laboratory practice purposes, from the early conventional (or monoplex) analytical methods able to detect single autoantibodies to the more recent multiplex platforms that can quantify tens of molecules. Although it has been in use for over 50 years, indirect immunofluorescence remains the standard method for research on many types of autoantibodies, due to its characteristics of diagnostic sensitivity and also to recent technological innovations which permit it a greater level of automation and standardization. The recent multiplex immunometric methods, with varying levels of automation, present characteristics of higher diagnostic accuracy, but are not yet widely diffused in autoimmunology laboratories due to the limited number of autoantibodies that are detectable, and due to the high cost of reagents and systems. Technological advancement in autoimmunology continues to evolve rapidly, and in the coming years new proteomic techniques will be able to radically change the approach to diagnostics and possibly also clinical treatment of autoimmune diseases. The scope of this review is to update the state of the art of technologies and methods for the measurement of autoantibodies, with special reference to innovations in indirect immunofluorescence and in multiple proteomic methods.


Banfi G.,University of Milan | Lundby C.,University of Zürich | Robach P.,Ecole Nationale de Ski e dAlpinisme | Lippi G.,Laboratorio Analisi
European Journal of Applied Physiology | Year: 2011

The influence of training and competition workloads is crucial for evaluation of longitudinal haematological data in athletes. There are only a few papers on the variation of haematological parameters during long-lasting periods and, especially, during an entire competitive season. We summarized that some haematological parameters can be influenced by long-term training and competition periods. Haemoglobin (Hb) and haematocrit (Ht) are decreased during the more intense periods of training, throughout the season. In different sport disciplines, the decline of Hb ranges from 3 to 8% during the competition season, while the range of reticulocytes (Ret%) varies from 5 to 21%. Reticulocytes are also decreased after long periods of training and competitions, but their variation is not necessarily associated with that of Hb. The qualitative variations (trend of modifications) of haematological parameters are roughly independent of the sport discipline, but quantitatively (amount of modifications) dependent on sport discipline. The modifications are more evident in cycling, running, swimming than they are in football and rugby. The variations of haematological parameters within the same sport discipline are qualitatively concordant and quantitatively different among separate but consecutive competitive seasons. These findings are described in aerobic and team sports sportsmen. The definition of reliable reference ranges in sportsmen would only be possible by following the best laboratory practices. For antidoping purposes more studies investigating haematological modifications during the season are advisable. © 2010 Springer-Verlag.


Bizzaro N.,Laboratorio Of Patologia Clinica | Antico A.,Laboratorio Analisi | Platzgummer S.,Laboratorio Centrale | Tonutti E.,Immunopatologia e Allergologia | And 5 more authors.
Autoimmunity Reviews | Year: 2014

Background: Indirect immunofluorescence (IIF) plays an important role in immunological assays for detecting and measuring autoantibodies. However, the method is burdened by some unfavorable features: the need for expert morphologists, the subjectivity of interpretation, and a low degree of standardization and automation. Following the recent statement by the American College of Rheumatology that the IIF technique should be considered as the standard screening method for the detection of anti-nuclear antibodies (ANA), the biomedical industry has developed technological solutions which might significantly improve automation of the procedure, not only in the preparation of substrates and slides, but also in microscope reading. Methods: We collected 104 ANA-positive sera from patients with a confirmed clinical diagnosis of autoimmune disease and 40 ANA-negative sera from healthy blood donors. One aliquot of each serum, without information about pattern and titer, was sent to six laboratories of our group, where the sera were tested with the IIF manual method provided by each of the six manufacturers of automatic systems. Assignment of result (pos/neg), of pattern and titer was made by consensus at a meeting attended by all members of the research team. Result was assigned if consensus for pos/neg was reached by at least four of six certifiers, while for the pattern and for the titer, the value observed with higher frequency (mode) was adopted. Seventeen ANA-positive sera and six ANA-negative sera were excluded. Therefore, the study with the following automatic instrumentation was conducted on 92 ANA-positive sera and on 34 ANA-negative sera: Aklides, EUROPattern, G-Sight (I-Sight-IFA), Helios, Image Navigator, and Nova View. Analytical imprecision was measured in five aliquots of the same serum, randomly added to the sample series. Results: Overall sensitivity of the six automated systems was 96.7% and overall specificity was 89.2%. Most false negatives were recorded for cytoplasmic patterns, whereas among nuclear patterns those with a low level of fluorescence (i.e., multiple nuclear dots, midbody, nuclear rim) were sometimes missed.The intensity values of the light signal of various instruments showed a good correlation with the titer obtained by manual reading (Spearman's rho between 0.672 and 0.839; P. <. 0.0001 for all the systems). Imprecision ranged from 1.99% to 25.2% and, for all the systems, it was lower than that obtained by the manual IIF test (39.1%). The accuracy of pattern recognition, which is for now restricted to the most typical patterns (homogeneous, speckled, nucleolar, centromere, multiple nuclear dots and cytoplasmic) was limited, ranging from 52% to 79%. Conclusions: This study, which is the first to compare the diagnostic accuracy of six systems for automated ANA-IIF reading on the same series of sera, showed that all systems are able to perform very well the task for which they were created. Indeed, cumulative automatic discrimination between positive and negative samples had 95% accuracy. All the manufacturers are actively continuing the development of new and more sophisticated software for a better definition in automatic recognition of patterns and light signal conversion in end-point titer. In the future, this may avert the need for serum dilution for titration, which will be a great advantage in economic terms and time-saving. © 2013 Elsevier B.V.


Righetti G.,Laboratorio Analisi | Graziani M.S.,Laboratorio Analisi
Biochimica Clinica | Year: 2011

The IgD monoclonal gammopathy is a rare event and its recognition and management are quite important because the condition is potentially life-threatening. This paper reports on six cases of IgD monoclonal components with emphasis on diagnostic procedures to be carried out in the protein laboratory in order to correctly identify this plasma cell disorder. The usual tests for monoclonal gammopathies are not fully adequate to accurately diagnose this gammopathy type and special procedures should be undertaken.


Grillo C.,Laboratorio Analisi | Patrucco G.,Laboratorio Analisi
Biochimica Clinica | Year: 2010

The glomerular filtration rate (GFR) is considered the best index of kidney function, but cannot easily be measured in clinical practice. Serum creatininebased equations, such as the Modification of Diet in Renal Disease study (MDRD) equation, are, therefore, routinely used to estimate GFR. To improve the accuracy of GFR estimate clinical laboratories should use creatinine assays traceable to the isotope dilution-mass spectrometry (IDMS) reference method. This paper illustrates the results of a regional survey on the creatinine standardization and reporting of GFR estimate. The survey shows the need for an improvement in reporting and interpreting serum creatinine and GFR calculation.

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