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This study investigated the biological effects on circulating monocytes after challenge with SARS-CoV-2 recombinant spike protein. Whole blood collected from seven ostensibly healthy healthcare workers was incubated for 15 min with 2 and 20 ng/mL final concentration of recombinant spike protein of Ancestral, Alpha, Delta, and Omicron variants. Samples were analyzed with Sysmex XN and DI-60 analyzers. Cellular complexity (i.e., the presence of granules, vacuoles and other cytoplasmic inclusions) increased in all samples challenged with the recombinant spike protein of the Ancestral, Alpha, and Delta variants, but not in those containing Omicron. The cellular content of nucleic acids was constantly decreased in most samples, achieving statistical significance in those containing 20 ng/mL of Alpha and Delta recombinant spike proteins. The heterogeneity of monocyte volumes significantly increased in all samples, achieving statistical significance in those containing 20 ng/mL of recombinant spike protein of the Ancestral, Alpha and Delta variants. The monocyte morphological abnormalities after spike protein challenge included dysmorphia, granulation, intense vacuolization, platelet phagocytosis, development of aberrant nuclei, and cytoplasmic extrusions. The SARS-CoV-2 spike protein triggers important monocyte morphological abnormalities, more evident in cells challenged with recombinant spike protein of the more clinically severe Alpha and Delta variants.

Chronic anaemia in advanced liver disease is a frequent finding. The aim was to explore the clinical impact of spur cell anaemia, a rare entity typically associated with end-stage of the disease. One-hundred and nineteen patients (73.9% males) with liver cirrhosis of any etiology were included. Patients with bone marrow diseases, nutrients deficiencies and hepatocellular carcinoma were excluded. In all patients, a blood sample was collected to check for the presence of spur cells on blood smear. A complete blood biochemical panel was recorded together with Child–Pugh (CP) score and Model for End-Stage Liver Disease (MELD) score. For each patients, clinically relevant events, such as acute-on-chronic liver failure (ACLF) and 1 year liver-related mortality, were registered. Patients were then grouped according to the percentage of spur cells at smear (> 5%, 1–5%, 5% spur cells, only 2 had evidence of haemolysis. In patients with > 5% spur cells, haemoglobin and albumin were lower compared with the other sub-group, while MELD score, CP score, International Normalized Ratio, ferritin, creatinine and unconjugated bilirubin were higher. Patients with more spur cells were more decompensated and developed more frequently ACLF. ACLF and liver-related mortality were significantly and independently associated with the presence of > 5% spur cells but not with baseline severe anaemia. Cirrhotic patients have a fairly high prevalence of spur cells, not always associated with severe haemolytic anaemia. The presence of spur red cells is per se associated with a worse prognosis and, therefore, should be always evaluated to prioritize patients for intensive management and eventually liver transplantation.

AimsOptical microscopic (OM) evaluation of peripheral blood (PB) cells is still a crucial step of the laboratory haematological workflow. The morphological cell analysis is time-consuming and expensive and it requires skilled operator. To address these challenges, automated image-processing systems, as digital morphology (DM), were developed in the last few years. The aim of this multicentre study, performed according to international guidelines, is to verify the analytical performance of DM compared with manual OM, the reference method.MethodsFour hundred and ninety PB samples were evaluated. For each sample, two May Grunwald-stained and Giemsa-stained smears were performed and the morphological evaluation of cells was analysed with both DM and OM. In addition, the assessment times of both methods were recorded.ResultsComparison of DM versus OM methods was assessed with Passing-Bablok and Deming fit regression analysis: slopes ranged between 0.17 for atypical, reactive lymphocytes and plasma cells (LY(AT)) and 1.24 for basophils, and the intercepts ranged between −0.09 for blasts and 0.40 for LY(AT). The Bland-Altman bias ranged between −6.5% for eosinophils and 21.8% for meta-myemielocytes. The diagnostic agreement between the two methods was 0.98. The mean of assessment times were 150 s and 250 s for DM and OM, respectively.ConclusionDM shows excellent performance. Approximately only 1.6% of PB smears need the OM revision, giving advantages in terms of efficiency, standardisation and assessment time of morphological analysis of the cells. The findings of this study may provide useful information regarding the use of DM to improve the haematological workflow.

Introduction Recent automated hematology analyzers (HAs) can identify and report nucleated red blood cells (NRBC) count as a separate population out of white blood cells (WBC). The aim of this study was to investigate the analytical performances of NRBC enumeration on five top of the range HAs. Methods We evaluated the within-run and between-day precision, limit of blank (LoB), limit of detection (LoD), and limit of quantitation (LoQ) of XE-2100 and XN-module (Sysmex), ADVIA 2120i (Siemens), BC-6800 (Mindray), and UniCel DxH 800 (Beckman Coulter). Automated NRBC counts were also compared with optical microscopy (OM). Results The limits of detection for NRBC of the BC-6800, XN-module, XE-2100, UniCel DxH 800, and ADVIA 2120i are 0.035×109/L, 0.019×109/L, 0.067×109/L, 0.038×109/L, and 0.167×109/L, respectively. Our data indicated excellent performance in terms of precision. The agreement with OM was excellent for BC-6800, XN-module, and XE-2100 (Bias 0.023, 0.019, and 0.033×109/L, respectively). ADVIA 2120i displayed a significant constant error and UniCel DxH 800 both proportional and small constant error. Conclusion Regards to NRBC counting, the performances shown by BC-6800, XN-module, and XE-2100 are excellent also a low count, ADVIA 2120i and UniCel DxH 800 need to be improved.

Immune Thrombocitopenic Purpura (ITP) is an autoimmune disease characterized by antibody-mediated platelet destruction and variable reduced platelet production. Besides antibody-mediated platelet destruction, new pathogenic mechanisms have been reported to be involved in reducing platelet count. Among these, desialylation is one of the most recent and innovative mechanisms that has been found to be implied, at least in part, in non-antibody mediated platelet clearance. Common Variable Immunodeficiency (CVID) is the most common Primary Immunodeficiency seen in clinical practice. About 25-30% of CVID patients are affected by autoimmune manifestation, among which ITP is the most common. Little is know about pathophysiological mechanisms that lead to ITP in CVID. Given the poor antibody production typical of CVID patients, we aimed at verifying whether platelet desialylation could be responsible for CVID associated thrombocytopenia. According to our results, we may suggest that in CVID patients, ITP is due to a decreased bone marrow platelets production, rather than an increased peripheral platelet destruction, which is more common in patients with primary ITP. An increased platelet desialylation does not appear to be implicated in the thrombocytopenia secondary to CVID, while it is implicated in the pathogenesis of primary ITP. Nevertheless an intriguing aspect has emerged from this study: regardless the presence of thrombocytopenia, the majority of CVID patients present a double platelet population as far as desialylation concerns, whilst no one of the healthy donors and of the patients with primary ITP shows a similar characteristic.

Supplemental Digital Content is available in the text., Objective: Pulmonary thrombosis is observed in severe acute respiratory syndrome coronavirus 2 pneumonia. Aim was to investigate whether subpopulations of platelets were programmed to procoagulant and inflammatory activities in coronavirus disease 2019 (COVID-19) patients with pneumonia, without comorbidities predisposing to thromboembolism. Approach and Results: Overall, 37 patients and 28 healthy subjects were studied. Platelet-leukocyte aggregates, platelet-derived microvesicles, the expression of P-selectin, and active fibrinogen receptor on platelets were quantified by flow cytometry. The profile of 45 cytokines, chemokines, and growth factors released by platelets was defined by immunoassay. The contribution of platelets to coagulation factor activity was selectively measured. Numerous platelet-monocyte (mean±SE, 67.9±4.9%, n=17 versus 19.4±3.0%, n=22; P

Background The osmotic fragility test (OFT), conventionally used for assisting the diagnosis of many erythrocyte disorders, is a manual and time-consuming analysis not daily performed in many medical laboratories. This study was aimed at defining the stability of whole blood samples used for assessing erythrocyte osmotic resistance. Methods Twenty-one consecutive routine whole blood samples collected into 5.4 mg K2EDTA were tested immediately after collection (day 0) and at different time intervals afterward (day 1, 2, 3, 4, 7, 10 and 14) after storage at 4 °C. The OFT was performed with the Osmored Monotest (1.3% glycerol; Eurospital, Trieste, Italy). Results at the different time points were compared with those obtained at day 0 and with the reference change value (i.e., 33%). Results The median value of both hyperosmolar and hyposmolar resistance increased from baseline, reaching statistical significance at day 7 for hyperosmolar resistance and at day 1 for hyposmolar resistance, respectively. The median relative increase of hemolysis percentage values become greater than the reference change value at day 3 for hyposmolar resistance, while this limit was never overcome for hyperosmolar resistance. A significant inverse association was found between the mean increase in hyperosmolar resistance and the baseline value of hyperosmolar resistance (r = −0.92), mean corpuscular volume (MCV; r = −0.46) or mean corpuscular hemoglobin (MCH; r = −0.44), as well as between the mean increase in hyposmolar resistance and the baseline value of hyposmolar resistance (r = −0.86), or patient age (r = −0.56). Conclusions The sample stability seems critical for the OFT. Whole blood specimens should not be stored refrigerated at 4 °C for >2 days before testing.

Background Ethylenediaminetetraacetic acid (EDTA)-dependent pseudothrombocytopenia is a rare phenomenon. Spurious pseudothrombocytopenia has also been described in other circumstances, while artifactual platelet count in whole blood samples anticoagulated with sodium citrate is an exceptional occurrence. Case report In this study, we describe the case of a 44-year-old ostensibly healthy woman who attended the local outpatient clinic for routine laboratory testing, including platelet count in EDTA and sodium citrate, for suspected artifactual pseudothrombocytopenia previously identified in another center. The results of hematological testing on both specimens were essentially normal, except for mild anemia. Nevertheless, the platelet number was 425 × 109/L in K2EDTA and 266 × 109/L (293 × 109/L after correcting for sample dilution) in sodium citrate, respectively. Microscopic revision of blood smears revealed the presence of platelet aggregates and satellitism only in the sodium citrate specimen. Conclusion Unlike previous occasional reports of concomitant EDTA- and sodium citrate-dependent pseudothrombocytopenia, we first describe a paradigmatic case of artifactual platelet count attributable to platelet clumping and satellitism, exclusively developing in blood anticoagulated with sodium citrate.

BACKGROUND: Successful peripheral blood stem cell (PBSC) collection depends on optimal timing of apheresis, as usually determined by flow cytometry CD34-positive ((+)) cell count in peripheral blood (PB). Since this method is costly and labour-intensive, we evaluated the use of the Hematopoietic Progenitor Cell count programme on a Sysmex(®) XN haematologic analyser (XN-HPC) as a rapid and inexpensive alternative for predicting CD34(+) cell count in PB samples. MATERIALS AND METHODS: Haematopoietic progenitor cell and CD34(+) cell counts were compared using 273 PB samples collected from 78 healthy donors and 72 patients who underwent PBSC transplantation. We assessed the effectiveness of the XN-HPC in safely predicting pre-harvest CD34(+) counts. The most efficient cut-off values of XNHPC were identified. We also evaluated the imprecision (coefficient of variation, CV) and functional sensitivity. RESULTS: Imprecision of the XN-HPC count was

Background The aims of this study were to compare the diagnostic accuracy of blood smear review criteria, by means of three different panel rules, those proposed by: the International Consensus Group for Hematology [41-ICGH rules], the Italian Survey [IS rules] and the Working Group on Hematology-SIBioC (WGH) consensus rules (WGH rules). Methods This study is based on 2707 peripheral blood (PB) samples referred for routine hematological testing to the WGH-associated laboratories displaced all over the Italian territory. The PB samples were processed on seven different hematology analyzers (HAs): Advia 2120i, XE-2100, BC-6800, ABX Pentra, XN-1000, Cell-DYN Sapphire, and DxH800, respectively. All the results provided by the HAs were analyzed through the application of three different blood smear review criteria: that is, the 41-ICGH, IS, and WGH rules. Finally, data were compared with those obtained by optical microscopy (OM), as the current gold standard. Results The overall the agreement OM classification with ICGH, IS, and WGH panel rules is 0.83, 0.83, and 0.85, respectively. The false negatives are 2.1%, 3.0%, and 2.9%, while false positives are 15.1%, 13.7%, and 11.7%, respectively. All the seven HAs showed variable interinstrument performance, as three different criteria for OM review were adopted on each of them from time to time. Conclusion These results presented show that the customization of validation rules is necessary for enhancing the quality of hematological testing and optimizing workflow.

Background Acute myocardial infarction (AMI; ICD9-CM 410*) is a leading cause of morbidity and mortality all over the world, and its community surveillance is essential to monitor variation in the occurrence of the disease. Between the late 1990s and the early 2000s more sensitive and specific biomarkers of myocardial necrosis (ie, troponins) were introduced and new diagnostic criteria, emphasising the role of biomarkers, have been developed for clinical and epidemiological purposes. Methods Tosc-AMI is a population-based registry based on the record linkage between hospital and mortality databases; it provides trends of coronary events in Tuscany, Italy. Two random samples of patients admitted to hospital in 2003 were validated according to the American Heart Association (AHA; 2003) and the Multinational MONItoring of trends and determinants in CArdiovascular disease (MONICA) (1983) criteria. Sample 1 (380 cases) was represented by patients admitted to hospital for AMI and sample 2 (380 cases) for other coronary diagnosis. Results Tosc-AMI attack rates increased from the period 1997 to 2005 (men: +17%; women: +30%) and then they decreased in the following 2 years (men: −8%; women: −13%). The rise of AMI hospital admissions was due to cases with ICD9-CM code 410.7 (largely representing non-ST elevation MI). According to the AHA criteria, 94.6% events of sample 1 and 29.8% events of sample 2 fulfilled the most extensive criteria for definite, probable or possible AMI. As expected, the more updated AHA definition identified as definite AMI an additional 33.3% when compared to the MONICA criteria (86.0% vs 52.7%). Conclusions The study suggests an influence of the new diagnostic criteria on the rising AMI trend observed in the early 2000s, an increase of less severe cases and a decreasing trend of forms with a more extended myocardial damage.

Peroxiredoxin-2 (Prx2), a typical 2-cysteine (Cys) peroxiredoxin, is a key anti-oxidant system in both normal and pathological erythropoiesis characterized by oxidative stress such as b-thalassemia. We recently showed that the absence of Prx2 worsens ß-thalassemic erythropoiesis and related iron-overload (Matte A et al. Antioxid Redox Signal, 2015). Here, we studied the effects of iron overload in a mouse model genetically lacking Prx2 (Prx2-/-). Two months old female wild-type (WT) and Prx2-/- pure bred mice were fed with a diet containing 2.5% carbonyl-iron compared to standard diet treated mice. We evaluated hematologic parameters, red cell indices and reticulocyte count in both mouse strains at baseline and at 30, 49, 60 and 90 days of treatment with carbonyl-iron. We observed a rapid drop in Hct and reticulocyte count in Prx2-/- mice compared to wild-type mice between 30-49 days of iron supplementation with the appearance of severe hyporegenerative anemia at 60 days of treatment in Prx2-/- characterized by a significant reduction in CD44+TER119+Fsc high cells. This was associated with marked increases in apoptotic Prx2-/- orthochromatic erythroblast compared to either baseline values or WT treated mice. In sorted erythroid precursors from iron overload WT mice, Prx2 expression was significantly increased compared to WT under standard diet. We observed a modulation of Erythropherrone (Erfe) expression during erythropoiesis with upregulation of Erfe in WT orthochromatic erythroblast compared to Prx2-/- erythroblasts. In liver from Prx2-/- mice exposed to iron-overload, we found liver iron content similar to WT mice but Pearls stain analysis showed differential iron distribution in cellular components of liver. While iron accumulated in hepatocytes and Kuppfer cells in Prx2-/- mice, iron-deposits were present only in hepatocytes of WT liver. Oxyblot analysis and liver MDA levels were significantly higher in Prx2-/- mice indicating that the absence of Prx2 promotes a severe liver oxidative stress. In WT liver, Prx2 expression was marked increased in a time depend way during iron supplementation, indicating that Prx2 is part of an adaptive cellular response to iron overload. This is in agreement with increased levels of ferritin-H in Prx2-/- mice compared to WT mice. Hepcidin (HAMP) expression was markedly increased in iron-overload WT mice compared to untreated control group, while no major changes were observed in Prx2-/- mice. Tfr2 expression was significantly increased only in livers of iron-overload WT mice, whereas phospho smad 1-5 was significantly increased in both mouse strains in response to iron-overload. The activation of the signaling pathway through Erk-1/2 only in iron-overload WT mice but not in Prx2-/- mice is most likely related to severe oxidative stress in Prx2-/- resulting in switching off of the Erks pathway. Importantly, administration of PEP1-Prx2 fusion protein rescue almost completely the hematologic phenotype with modulation of Erk signaling pathway towards Tfr2 and the smad system, validating our hypothesis of a role for Erk signaling for the observed phenoytpes. Our data highlight Prx2 as novel factor involved in iron homestasis through the control of oxidative stress modulating signaling pathway towards hepcidin expression. Disclosures No relevant conflicts of interest to declare.

Background: The number of elderly persons with cardiovascular disease (CVD) is rapidly increasing, mainly due to worldwide aging population. All CVD risk scores consider age as risk factor but most of them assume that the etiological role of other established risk factors is similar at all ages. This analyses aimed to assess the predictive role of cardiovascular risk factors in elderly persons. Methods: We considered 10,979 men and 16,533 women aged 35-74 years from cohorts in North, Centre and South of Italy with 10 years median follow-up time for fatal and non fatal coronary and cerebrovascular events (819 in men and 467 in women) validated according to MONICA criteria. In all cohorts risk factors were assessed by using standardized procedures. Two age groups were defined: adult 35-54 years of age (6,353 men and 10,358 women) and elderly 55-74 years (4,626 men and 6,175 men). Cox models were implemented including age, systolic blood pressure (SBP), total (TC) and HDL cholesterol (HDL-C), smoking, diabetes (DM - fasting blood glucose >=126 mg/dl or under glucose treatment) and hypertension treatment (HT). For continuous variables, hazard ratios (HR) were reported at 1 standard deviation higher level and for dichotomized variables, yes vs no. Results: For adult men SBP resulted having a stronger impact on CVD risk than for elderly even if not statistically significant: adult HR 1.32, 95% confidence interval 1.18-1.48, vs elderly HR 1.25, 1.15-1.36; similar results for TC (HR 1.47 vs 1.25), smoking (HR 1.91, vs 1.52), DM (HR 1.76 vs 1.56), HT (HR 2.30 vs 1.25). HDL-C resulted not significantly associated with CVD risk in elderly men. Among adult women a stronger impact was found than in elderly for SBP (HR 1.59 vs 1.26), HDL-C (0.65 vs 0.84), smoking (2.43 vs 1.99), and HT (2.34 vs 1.45); the opposite for TC (1.19 vs 1.27). DM resulted not significantly associated with CVD risk in adult women. Age resulted having a stronger impact in elderly persons both in men and women (HR 1.22, 1.07-1.39, vs 1.44, 1.29-1.61; 1.45, 1.17-1.80, vs 1.63, 1.42-1.86 respectively). Conclusions: SBP and HT resulted having a pronounced lower impact on CVD risk in elderly than in adult as well as HDL-C for men; age shows a stronger impact in elderly than in adult. Further analyses are recommended to study the reliability of risk scores based on age-specific risk models, evaluating the introduction of different risk factors and considering survival probability derived from data in elderly population.

The preanalytical phase is considered the most vulnerable phase in biopreservation, biobanking, and laboratory diagnostics. Accurate mixing after blood collection is claimed to be important and recommended by the manufacturers.To evaluate whether it is really necessary to mix the primary blood tubes immediately after blood collection by means of evacuated tube systems.Blood from 300 outpatients was equally and randomly divided into three groups: G1, sodium citrate vacuum tubes; G2, lithium heparin vacuum tubes; and G3, K2EDTA vacuum tubes. All vacuum tubes were processed using three different procedures. Procedure 1: Gold Standard (P1): All specimens mixed gently and carefully by inverting five times as recommended; Procedure 2: Rest time (P2): All specimens remained 5 min in the upright position, followed by gentle careful mixing by inverting five times; Procedure 3: No mix (P3): All specimens were left in upright position without mixing afterwards. The influence of the primary mixing tube procedure was evaluated for clinical chemistry, hematology, and coagulation parameters by paired t-test. The bias from the mixing procedure was also compared with quality specifications derived from biological variation.Significant differences (p0.017) were found for: i) red blood cell count and hematocrit when P1 was compared with P2; ii) alanine aminotransferase and erythrocyte sedimentation rate when P1 was compared with P3; iii) red blood cell count, hematocrit, and hemolysis index when P2 was compared with P3. Surprisingly, clinically significant differences were found only for sodium when P1 was compared with P2, and P1 was compared with P3. No fibrin filaments or microclots were observed in any samples.Primary blood tubes mixing after collection with evacuated tube system appears to be unnecessary.

Heat-killed vegetative forms of Bacillus subtilis were found to impair considerably the capacity of human T-lymphocytes to secrete interleukin-2 (IL-2) and to proliferate (in terms of [3H]thymidine incorporation) after phytohaemagglutinin (PHA) stimulation. B. subtilis was also found to interfere with T-cell proliferation induced by concanavalin A (Con A) and the recall antigen tetanus toxoid (TT). The suppressive activity was dependent on bacterial concentration, and was not ascribed to mitogen, medium-nutrient absorption or cell killing. Moreover, B. subtilis did not interfere with mitogen-induced IL-2 receptor expression on the T-cell surface. On the other hand, B. subtilis did not interfere with T-cell proliferation induced by phorbol myristate acetate (PMA) and ionomycin stimulation. All data obtained suggest the binding of B. subtilis subcomponents to- or very close to-the T-cell receptor (TCR). Identification and purification of the basic structure(s) or component(s) of B. subtilis with TCR antagonist activity in vitro will help to exploit different aspects of T-cell activity and development, and possibly, will provide a means of specific control or modification of the immune response.

Background: From 2008 to 2012 an Health Examination Survey (HES)-Osservatorio Epidemiologico Cardiovascolare (OEC) has been implemented in Italy with the aim of assessing cardiovascular (CV) risk factors, prevalence of risk conditions and CV diseases for the Italian adult population. Methods: Random samples of general population stratified by age and sex were examined in all Italian regions (participation rate 56%). Risk factor were collected using standardized procedures and methods; biochemical tests were assayed in a central laboratory; a questionnaire investigates behaviours and CVD history; a ECG read in Minnesota code was used to define previous myocardial infarction. Comparisons between men and women were assessed using t-test for means and chi-squared test for prevalence. Results: Data of 4371 men and 4339 women ages 35-79 years were analysed. Majority of risk factors mean levels resulted higher in men than in women: systolic blood pressure (SBP) was 134 mmHg and 129 mmHg (p=140 mmHg or DBP>=90 mmHg or in treatment (p=200 mg/dl or in treatment for (p2 ) (p=30 kg/m 2 ) resulted similar in men and women: 25% and 27% respectively (p=0.0818). Prevalence of myocardial infarction was 2.1% in men and 0.7% in women (p Conclusions: At present obesity and smoking are still a priority in public health. In combination with other information sources, the OEC can contribute greatly to plan community actions and health services at national and regional level.

Background: Health Interview Survey (HIS) collects only self-reported height and weight to estimate obesity prevalence. Because of self-reporting, estimated obesity prevalence could be too low. From 2008 to 2012 an Health Examination Survey/Osservatorio Epidemiologico Cardiovascolare (HES/OEC) has been implemented in Italy with the aim of assessing cardiovascular (CV) risk factors, prevalence of risk conditions and CV diseases for the Italian adult population. Differences in measured and self-reported main anthropometric measures are described. Methods: Random samples of general population stratified by age and sex were examined in all Italian regions (participation rate 56%). Self-reported height and weight were requested to participants just after the welcome at the screening center. Height and weight were then measured using standardized procedures and methods. BMI was categorized in 3 groups: normal (BMI=30 kg/m2). Comparisons between measured and reported height, weight, and calculated BMI in men and women were assessed using t-test for means and chi-squared test for prevalence for paired samples. Results: Data from 2,583 men and 2,575 women aged 35-79 years were analysed. In men, measured height was lower than self-reported (-2.1 cm.), measured weight resulted higher than self-reported (+0.3 kg.); as a consequence BMI calculated on measured data resulted higher than that based on self-reported data (+0.8 kg/m2). Prevalence of obesity was higher for measured data than for self-reported (+7.1%). In men, difference between measured and self-reported height resulted higher in obese category than in the overweight or normal groups (-2.6 cm.), as well as for weight (+1.6 kg.). Similar results were registered in women: measured height was lower than self-reported (-3.6 cm.), measured weight resulted higher than self-reported (+0.8 kg.); as a consequence BMI calculated on measured data resulted higher than that based on self-reported data (+1.6 kg/m2). Prevalence of obesity was higher for measured data than for self-reported (+9.2%). Difference between measured and self-reported height resulted higher in obese category than in the overweight or normal groups (-5.0 cm.), as well as for weight (1.8 kg.). Both in men and women differences increased by age-group. Conclusions: Our results confirm how important is to produce standardised measures, also to adjust and correct self-reported height and weight, in order to have a reliable picture of obesity in the population. At present obesity is still a priority in public health and is one of the main indicator to plan community actions for cardiovascular disease prevention in the population.

Background: Despite cardiovascular diseases (CVD) are the leading causes of death and hospitalization in nearly all countries in Europe, there are paucity, weak quality and comparability of data. The periodic Health Examination Survey (HES) represents in Italy the major source of information on CVD at national level thanks to the adoption of standardized methodologies throughout the country. The aim is to present differences in trends of CVD prevalence by gender in the Italian adult population from 1998 to 2008. Methods: Randomized population samples stratified by age and sex were examined. In 1998, 9,712 men and women aged 35–74 years were enrolled in all Italian regions; ongoing screening started in 2008 is enrolling 9,020 persons. A standardized questionnaire investigates cardiovascular disease and pharmacological treatments. The anamnesis is positive when clinical diagnosis has been made by a physician. Electrocardiograms read in Minnesota code are also performed and used to define previous myocardial infarction. Results: Data are from 12 regions in the North, Center, and South of Italy: 3,704 men and women in 1998 and 3,479 persons in 2008 are compared. In men, prevalence of cerebrovascular events decreases from 1.5% (95% confidence interval: 1.0-1.9%) in 1998 to 0.6 (0.3–0.9%) in 2008; in women, prevalence results stable: 0.9% in 1998 (0.5–1.3%) and 0.7% in 2008 (0.4–1.1%). Prevalence of myocardial infarction results stable both in men and women: in 1998, 1.9% in men (1.4–2.4%) and 0.6% in women (0.3–0.9%), and in 2008, 2.0% (1.4-2.7%) and 0.6% (0.3-0.9%) respectively; angina pectoris results stable as well: in 1998, 2.2% in men (1.7-2.8%) and 3.9% in women (3.1-4.6%), and in 2008, 3.2% (2.5-4.0%) and 4.5% (3.6-5.4%) respectively. Prevalence of by-pass or angioplasty surgery increases in men: 2.4% (1.8-3.0%) in 1998, and 4.5% (3.6-5.4%) in 2008; in women increasing is not significant: from 0.2% (0.0-0.4%) in 1998 to 0.7% (0.3-1.1%) in 2008. Stratifying data by geographical area (North, Center, South and Island) all comparisons become not significant; in men only, cerebrovascular decreasing and by-pass or angioplasty surgery increasing trends tend to be confirmed in all three areas. Conclusions: Trends between 1998 and 2008 decrease in cerebrovascular events and increase in prevalence of by-pass or angioplasty surgery, especially in men. Stable prevalence resulted for myocardial infarction and angina pectoris. Cerebrovascular decreasing trend is in line with smoking prevalence and mean of blood pressure decreasing in the country. Further analyses considering data from all Italian regions are needed to confirm results.

Background: The Italian National Prevention Plan 2005–2008 included a 10-year cardiovascular risk assessment (10-CR) for the general population aged 35–69 years using the CUORE-Project risk score. GPs were encouraged to collect data on risk factors, perform a 10-CR and send these data to the Cardiovascular Risk Observatory (CRO). Aim: The aim of this study is to show updated data of the ongoing surveillance system of the 10-CR in the Italian adult population, a first step to implement primary preventive actions at individual level. Methods: A training plan for GPs was launched by the Ministry of Health. Data were collected using the cuore.exe software, easily and freely downloadable by GPs from the CUORE-Project website ( www.cuore.iss.it ). The CRO provides a web-platform to analyze and compare data on 10-CR and risk factors at both regional and national level. In the subgroup of persons examined at least twice a year, variations in continuous risk factors mean levels and categorical risk factors prevalences between baseline and follow-up and their 95% confidence intervals (C.I.) were calculated using methods for matched pair samples. Results: By October 2011, more than 3,000 GPs downloaded cuore.exe ; 146,322 CR assessments on 137,773 persons were sent to CRO. CR mean was 3.0% in women, 8.4% in men; 30% of men and 65% of women were at lower risk (CR Conclusions: Data demonstrate that 10-CR assessment can be an effective first step to implement preventive actions in primary care. The individual risk score is becoming a useful tool for GPs to assess their patients’ CR and promote primary prevention by focusing attention on the adoption of healthy lifestyles. These encouraging data can be used to support health policy decision processes.

The Italian national prevention plan 2005-2008 included 10-year cardiovascular risk (10-CR) assessment of the general population aged 35-69 years using the CUORE project risk score. General practitioners (GPs) were encouraged to collect data on risk factors and 10-CR and to contribute to the Cardiovascular Risk Observatory (CRO). The aim is to demonstrate feasibility and effectiveness of 10-CR assessment as a first step to implement primary preventive actions at the individual level.Data were collected using CUORE.EXE software, easily and freely downloadable by GPs from the CUORE project website (www.cuore.iss.it). CRO provides a web platform to analyse and compare data on 10-CR and risk factors at regional and national levels with the aim of supporting health policy decision processes.From January 2007 to May 2010, 2,858 GPs downloaded cuore.exe; 139,269 CR assessments on 117,345 persons were sent to CRO. CR mean was 3.0% in women, 8.3% in men; 30% of men and 65% of women were at lower risk (CR 3%), 9.2% of men and 0.4% of women were at high risk (CR ≥ 20%). Among those with at least two risk assessments (n = 5,948), 8% (95% CI 7-9%) shifted to a lower risk class after 1 year. Systolic blood pressure mean levels decreased by 1.6 mmHg (95% CI 1.2-2.1 mmHg), diastolic blood pressure by 0.9 mmHg (95% CI 0.5-1.3 mmHg), total cholesterol by 5.6 mg/dl (95% CI 4.3-6.8 mg/dl), and smokers prevalence by 3.5% (95% CI 2.5-4.6%); high-density lipoprotein cholesterol increased in women by 1 mg/dl (95% CI 0.5-1.4 mg/dl).Data demonstrate that 10-CR assessment can be the first step to implement preventive actions in primary care.

The Italian National Prevention plan includes 10-year cardiovascular risk (CR) assessment of the Italian general population aged 35-69 years using the CUORE Project risk score. A national training program for general practitioners (GPs) was launched by the Ministry of Health in 2003. GPs were encouraged to collect data on risk factors and risk assessment and to contribute to the CUORE Project Cardiovascular Risk Observatory (CRO). The aim of this analysis is to demonstrate the feasibility and effectiveness of risk assessment in primary care.The cuore.exe software, free of charge for GPs and easily downloadable from the CUORE Project web site (www.cuore.iss.it), is the frame for the GP data collection. The CRO provides a platform to analyze data collected on risk assessment and risk factors, and compare results at regional and national level in order to support health policy makers in their decision process.From January 2007 to April 2009, 2858 GPs have downloaded the cuore.exe software; 102,113 risk assessments were sent to the CRO based on risk factors profile of 87,556 persons (3617 persons had more than 1 risk assessment). Mean level of CR was 3.1% in women and 8.4% in men; 30% of men and 65% of women were at low risk (CR3%), 9% of men and 0.4% of women were found at high risk (CRor = 20%). Among those with at least 2 risk assessments, 8% shifted to a lower class of risk after 1 year. Mean level of systolic and diastolic blood pressure decreased by about 1% in 1 year; total cholesterol more than 2%, and prevalence of smokers decreased by about 3% in the second risk assessment.These data demonstrate that risk assessment can be included as a first step of prevention in primary care. The CUORE Project individual score is expected to become an important tool for GPs to assess their patients' CR, to promote primary prevention, and to focus attention to healthy lifestyle adoption.

Background Also in Mediterranean countries, which are considered a low risk population for cardiovascular disease (CVD), the increase in body mass index (BMI) has become a public health priority. To evaluate the feasibility of a CVD and obesity surveillance network, forty General Practitioners (GPs) were engaged to perform a screening to assess obesity, cardiovascular risk, lifestyle habits and medication use. Methods A total of 1,046 women and 1,044 men aged 35–74 years were randomly selected from GPs' lists stratifying by age decade and gender. Anthropometric and blood pressure measurements were performed by GPs using standardized methodologies. BMI was computed and categorized in normal weight (BMI 18.5–24.9 kg/m2), overweight (BMI 25.0–29.9 kg/m2) and obese (BMI ≥ 30 kg/m2). Food frequency (per day: fruits and vegetables; per week: meat, cheese, fish, pulses, chocolate, fried food, sweet, wholemeal food, rotisserie food and sugar drink) and physical activity (at work and during leisure time) were investigated through a questionnaire. CVD risk was assessed using the Italian CUORE Project risk function. Results The percentage of missing values was very low. Prevalence of overweight was 34% in women and 50% in men; prevalence of obesity was 23% in both men and women. Level of physical activity was mostly low or very low. BMI was inversely associated with consumption of pulses, rotisserie food, chocolate, sweets and physical activity during leisure time and directly associated with consumption of meat. Mean value of total cardiovascular risk was 4% in women and 11% in men. One percent of women and 16% of men were at high cardiovascular risk (≥ 20% in 10 years). Normal weight persons were four times more likely to be at low risk than obese persons. Conclusion This study demonstrated the feasibility of a surveillance network of GPs in Italy focusing on obesity and other CVD risk factors. It also provided information on lifestyle habits, such as diet and physical activity.

Over the last decades a change in physical activity habits of the Italian population has been observed. The aim of this study was to compare occupational and leisure-time physical activity, to assess the relationship between physical activity and cardiovascular risk factors and the variation in cardiovascular risk profile measured 20 years apart in the Italian population.In two populations aged 35-69 years (MATISS 1984-1987: 2041 men, 2424 women; OEC 1998-2002: 4214 men, 4206 women) gender-specific levels of occupational (occupational physical activity [OPA]-light, OPA-moderate, OPA-heavy) and leisure-time (leisure-time physical activity [LTPA]-low, LTPA-moderate, LTPA-high) physical activity were compared. Relationship between physical activity and cardiovascular risk factors was evaluated in the two samples using logistic regression models adjusted for age, heart rate, educational level and other possible confounding variables in men and women separately. The 10-year cardiovascular risk was assessed in the two periods and in different physical activity strata using the Progetto CUORE risk score.Comparing the two studies, prevalence of OPA-low had decreased, whereas the prevalence of LTPA-low had increased. With increasing LTPA blood pressure, heart rate and glycemia decreased, whereas high-density lipoprotein-cholesterol increased with a positive effect on obesity, smoking habit and cardiovascular risk.Over the last 20 years, physical activity apparently changed: currently men and women dedicate more time to physical activity during leisure-time, and are less active at work. More active people have lower levels of major cardiovascular risk factors. Societal efforts are needed to increase physical activity levels in the population.

The aim of this study was to assess the 10-year cardiovascular risk categories using risk chart, recently set up by the National Institute of Health in the population examined by the Cardiovascular Epidemiologic Observatory.3745 men and 3664 women aged 40-69 years were classified into five risk categories (5 %; 5-10%; 10-15%; 15-20%;or = 20%) taking into account age, smoking habit, history of diabetes, systolic blood pressure, serum cholesterol and excluding those already under treatment for hypertension and hypercholesterolaemia or experienced a previous major cardiovascular event (1937 persons: 955 men, 982 women).Proportion of people estimated at risk in 10 yearsor = 20% is minimal in the youngest age range, increases in adulthood, duplicates in smokers and is higher in diabetics. In non-diabetic men that proportion varies between 3.4% in non-smokers and 5.6% in smokers. All women at risk are already under specific treatment.Cardiovascular Epidemiologic Observatory data allowed to assess the expected proportion of individuals at risk in 10 yearsor = 20%. Besides attention to high-risk individuals, preventive measures supporting a healthier lifestyle in the general population must be adopted, considering that it will produce the greatest number of events.

Type 2 diabetes is the most frequent form of diabetes in the adult population and is associated with an increasing risk of cardiovascular diseases. The objective of this study was to describe the prevalence and the state of control in an Italian population sample examined within the Osservatorio Epidemiologico Cardiovascolare study.The sample of this study consisted in 8972 subjects, men and women aged 35-74 years. A fasting capillary blood glucoseor = 126 mg/dl or being on antidiabetic treatment were the criteria used to define diabetes, while a fasting capillary blood glucose140 mg/dl was considered to represent effective treatment (controlled diabetes) in treated patients.Diabetes was present in 8.4% of men and 6% in women. The prevalence of glucose intolerance was 8.2% in men and 4.3% in women. Only 50.7% of diabetic men and 56.5% of diabetic women knew about their disease; among these people, 21% was on treatment with oral antidiabetics and/or insulin. Such a therapy was effective in 39.5% of the treated cases.The prevalence of diabetes as well as glucose intolerance was greater in men and in the regions of the Center and South of Italy. The state of control of diabetes cannot be considered satisfactory; women and older age groups, when prevention strategies are less important, appear to be under better care.

Objectives To verify if hand-grip performance in older men is a predictor of disability. Design Population-based prospective study. Setting A sample from the Italian rural cohorts of the FINE study (Finland, Italy, Netherlands Elderly), representative of the general population of elderly men surveyed in 1991 and 1995. Participants 140 men aged 71-91 years who reported no disability in performing activities of daily living (ADLs), instrumental activity of daily living (IADLs) and mobility activities at baseline examination and provided information on their functional status at follow-up 4 years later. Measurements Disability was defined as needing help in performing ADLs, IADLs and mobility. Hand-grip strength was evaluated at baseline by a mechanical dynamometer. Results After adjusting for potential confounding variables, a lower concentration of high-density lipoprotein cholesterol was the only factor predicting disability in men aged 76 years or younger and only reduced hand-grip strength predicted incident disability in men 77 years or older. Conclusion Poor hand strength as measured by hand-grip is a predictor of disability in older people. The hand-grip test is an easy and inexpensive screening tool to identify elderly people at risk of disability.

A 20-year follow-up study for all causes of death has been conducted on two groups of men aged 40-59 at entry in two rural communities of northern and central Italy on a total of 1,712 subjects representing 98.9% of defined demographic samples. Personal characteristics or risk factors usually studied for coronary heart disease (CHD), as measured at entry, have been related to the risk of dying in 20 years (600 cases). By means of multivariate analysis, 11 out of 33 considered characteristics were shown to be significant predictors of any cause of death. These were mean blood pressure, age, arm circumference (protective), early death of parents, cigarette smoking, forced expiratory volume (protective), CHD, arcus senilis, vital capacity (protective), xanthelasma, and serum cholesterol. Discrimination between cases and noncases was satisfactory with about 40% of deaths in the upper quintile of the estimated distribution of risk and less than 7% in the lowest quintile (relative risk = 6.20).

A total of 1712 men aged 40 to 59 years in two rural cohorts of northern and central Italy have been followed up for 25 years after an entry examination in 1960. Forty one individual characteristics have been considered as possible predictors of death in the next 25 years. After exclusion of 55 men with life threatening diseases (cardiovascular and cancer) and of 161 men because of missing measurements, 1495 men have been analysed for relation between entry factors and subsequent death (n = 670). Twelve factors eventually emerged as powerful predictors of future death: in hierarchical order, age, blood pressure, forced expiratory volume, cigarette smoking, xanthelasma, mother life-status, arm circumference, father life-status, shoulder-pelvis ratio, vital capacity, arcus senilis, and serum cholesterol. Discrimination as provided by logistic modelling placed 19.6% of all cases in the upper decile of the estimated risk, 36.8% in the upper quintile, 2.5% in the lowest decile, and 7.1% in the lowest quintile. Out of those located in the lowest decile of risk, 11.4% died within 25 years while the corresponding percentage in the upper decile was 87.3%. Use of the Cox model yielded slightly better coefficients than logistic function.

Two cohorts of men aged 49-59 at entry, representing cluster samples of two rural areas in Northern and Central Italy, for a total of 1712 subjects have been followed-up for 20 years within an epidemiological study originally designed for cardiovascular disease. After 20 years, only 41 men have been judged to have remained substancially healthy throughout the observation period, i.e. free from a number of major diseases. Univariate and multivariate analyses trying to predict the maintenance of health status showed that among 21 selected characteristics only the following one had a significant power: age, cigarette smoking (adverse effect) and vital capacity (favourable effect). A minor role was also played by the body mass index (adverse effect) and forced expiratory volume (favourable effect). Those who did not remain healthy exibited a greater increase in blood pressure and body mass index.

A pool of two Italian rural population samples made up of 1,712 men aged 40-59 at entry was studied in 1960 and than followed up for 25 years. The multivariate analysis of the first major coronary event using the Cox model showed, in men aged 40-59, 45-64 and 50-69, the significant predictive role of age, systolic blood pressure, serum cholesterol and cigarette smoking, but not of body mass index, without marked differences attributable to the aging process. Changes in systolic blood pressure, serum cholesterol and cigarette smoking occurring between year 0, 5 and 10 of follow-up, as defined by two different indicators, increased significantly the predictability of coronary events occurring between years 10 and 25 of follow-up when added to the model including the baseline factors. It is inferred that increases and decreases (even of relative nature) of the three major risk factors around the entry levels are associated with higher and lower levels of coronary risk in the 15 years after the changes have occurred.

The present investigation was aimed at evaluating the incidence and prediction of a first major cerebrovascular (fatal or nonfatal) event. The study population included the two Italian rural samples of the Seven Countries Study (namely, Montegiorgio and Crevalcore), accounting for a total of 1,712 men aged 40-59 years at entry and followed up for mortality and morbidity for 25 years. A number of individual variables measured at baseline, at the fifth year, and at the tenth year of follow-up and possibly related to cerebrovascular events were considered. Of the 1,709 subjects free from major cerebrovascular events at entry 171 developed a first major cerebrovascular event, but for the multivariate Cox model analysis only 1,572 subjects and 152 events were employed due to some exclusions for missing data. Systolic blood pressure, indexes of respiratory function (protective), and physical activity at work (protective) demonstrated significant predictive roles for all ages and all lengths of follow-up considered. Other factors (presence of arrhythmias, presence of arcus senilis, and skinfold thickness [protective]), significantly contributed to the prediction, but in only some models. Time-related changes in systolic blood pressure significantly improved the prediction of cerebrovascular events. The multivariate prediction performed in this report allowed the validation of three risk factors (systolic blood pressure, respiratory function indexes, and physical activity at work) whose predictive powers remain stable with aging. The need for further studies specifically aimed at discriminating hemorrhagic from thrombotic events is suggested.