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2. SARS-CoV-2 antibody responses before and after a third dose of the BNT162b2 vaccine in Italian healthcare workers aged ≤60 years: One year of surveillance

Author

Franzese, M., Coppola, L., Silva, R., Santini, Stefano Angelo, Cinquanta, L., Ottomano, C., Salvatore, M., Incoronato, M., Santini S. A. (ORCID:0000-0003-1956-5899), Franzese, M., Coppola, L., Silva, R., Santini, Stefano Angelo, Cinquanta, L., Ottomano, C., Salvatore, M., Incoronato, M., and Santini S. A. (ORCID:0000-0003-1956-5899)

Abstract

This study monitored the anti-spike-receptor-binding domain (RBD) and neutralizing antibodies induced by the Pfizer/BioNTech mRNA BNT162b2 vaccine in a cohort of 163 healthcare workers aged ≤60 years. We have taken advantage of two study groups, both of whom received the first two doses in the same time window, but Group 1 (54 HCWs) received the third dose 2 months before Group 2 (68 HCWs) did. The cohorts were monitored from the 12th day after the first vaccine dose up to 1 month after the third vaccine dose for a total of eight time points and about 1 year of surveillance (T1 = 12 days after the first dose; T2 = 10 days after the second dose; T3 = 1 month after the second dose; T4 = 3 months after the second dose; T5 = 4 months after the second dose; T6 = 5 months after the second dose; T7 = 7 months after the second dose; T8 = 1 month after the third dose for Group 1; T8* = 9 months after the second dose for Group 2; T9 = 1 month after the third dose for Group 2). The mean value of anti-spike antibodies decreased faster over time, but at T7, its decline was significantly slowed (T7 vs. T8*). After the third dose, the anti-spike titer rose about 34-fold (T7 vs. T8 and T8* vs. T9) and the booster improved the anti-spike titer by about three times compared with that of the second dose (T3 vs. T8 and T3 vs. T9), and no difference was noted between the two groups. The neutralizing titer was evaluated at T3, T7, T8, and T9. Anti-spike and neutralizing antibodies were found to be strongly correlated (r2 = 0.980; p < 0.001). At T3, 70% of the participants had a neutralizing antibody titer >91% of total anti-spike antibodies that increased to 90% after the third dose (T8 and T9). However, when the anti-spike titer reached its lowest value (T7), the neutralizing antibody levels decreased even further, representing only 44% of total anti-spike antibodies (p < 0.0001). Our findings show that the third vaccine dose improves the humoral response, but the wane of the a

Published
2022

12. Ronal and metabolic effects of insulin lispro in type 2 diabetic sublects with overt nephropathy

Author

Ruggenenti, P, Flores, C, Aros, C, Ene-Iordache, B, Trevisan, R, Ottomano, C, Remuzzi, G, Ruggenenti P, Flores C, Aros C, Ene-Iordache B, Trevisan R, Ottomano C, Remuzzi G, Ruggenenti, P, Flores, C, Aros, C, Ene-Iordache, B, Trevisan, R, Ottomano, C, Remuzzi, G, Ruggenenti P, Flores C, Aros C, Ene-Iordache B, Trevisan R, Ottomano C, and Remuzzi G

Abstract

OBJECTIVE - To assess whether the insulin analog lispro may antagonize the renal effects of IGF-1, a mediator of glomerular hyperfiltration involved in the progression of diabetic and nondiabetic chronic nephropathies. RESEARCH DESIGN AND METHODS - in a randomized crossover study, we compared the renal and metabolic responses to regular or lispro insulin (0.1 units/kg body wt) administered after a euglycemic clamp and 5 and 30 min before a standard meal to 11 type 2 diabetic patients with macroalbuminuria. RESULTS - Two- and four-hour postprandial changes (vs. preprandial euglycemia) in glomerular filtration rate (GFR) followed a significantly different trend (5.8 +/- 5.0 vs. -6.3 +/- 4.7, P < 0.05; and 11.0 +/- 6.8 vs. 0.7 +/- 5.1%, P < 0.05) after regular insulin and lispro injection, respectively. After lispro, postprandial GFR changes were negatively correlated (r = -0.48, P = 0.0001) with plasma insulin concentration. After regular insulin, renal plasma flow increased in parallel with a decrease in renal vascular resistances. Both changes were fully prevented by lispro. Postprandial blood glucose maximum concentration (278 +/- 16 vs. 240 +/- 16 mg/dl, P < 0.01) and area under the curve (79,381 +/- 19,237 vs. 72,810 +/- 16,2 11 mg/dl per min, P < 0,05) were significantly lower after insulin lispro than after regular insulin injection, respectively, despite comparable postprandial insulin profiles. Changes in total and gluconeogenic amino acids followed a similar trend. Changes in blood glucose and plasma amino acids did not correlate with concomitant changes in GFR. CONCLUSIONS - in overt nephropathy of type 2 diabetes, lispro prevents glomerular hyperfiltration and offsets the renal effects of meal or meal-associated hyperglycemia by mechanisms possibly related to IGF-1 antagonism.

Published
2003

14. Evaluation of nucleated red blood cell count by Sysmex XE-2100 in patients with thalassaemia or sickle cell anaemia and in neonates

Author

Buoro, S, Vavassori, M, Pipitone, S, Benegiamo, A, Lochis, E, Fumagalli, S, Falanga, A, Marchetti, M, Crippa, A, Ottomano, C, Lippi, G, Buoro, S, Vavassori, M, Pipitone, S, Benegiamo, A, Lochis, E, Fumagalli, S, Falanga, A, Marchetti, M, Crippa, A, Ottomano, C, and Lippi, G

Abstract

Background. Current haematology analysers have variable sensitivity and accuracy for counting nucleated red blood cells in samples with low values and in all those conditions characterised by altered sensitivity of red blood cells to the lysing process, such as in beta-thalassaemia or sickle-cell diseases and in neonates. The aim of our study was to evaluate the performance of the automated analyser XE-2100 at counting nucleated red blood cells in the above-mentioned three categories of subjects with potentially altered red blood cell lysis sensitivity and yet a need for accurate nucleated red blood cell counts. Materials and methods. We measured nucleated red blood cell count by XE-2100 in peripheral blood samples of 187 subjects comprising 55 patients with beta-thalassaemia (40 major and 15 traits), 26 sickle-cell patients, 56 neonates and 50 normal subject. Results were compared with those obtained by optical microscopy. Agreement between average values of the two methods was estimated by means of Pearson's correlation and bias analysis, whereas diagnostic accuracy was estimated by analysis of receiver operating characteristic curves. Results. The comparison between the two methods showed a Pearson's correlation of 0.99 (95% CI; 0.98-0.99; p<0.001) and bias of -0.61 (95% CI, -1.5-0.3). The area under the curve of the nucleated red blood cell count in all samples was 0.98 (95% CI, 0.96-1.00; p<0.001). Sub-analysis revealed an area under curve of 0.99 (95% CI, 0.98-1.00; p<0.001) for patients with thalassaemia, 0.94 (95% CI, 0.85-1.00; p<0.001) for patients with sickle cell anaemia, and 1.00 (95% CI, 1.0-1.0) for neonates. Discussion. XE-2100 has excellent performance for nucleated red blood cell counting, especially in critical populations such as patients with haemoglobinopathies and neonates.

Published
2015

21. Analytical comparison between two hematological analyzer systems: CAL-8000 vs. XN-9000.

Author

Buoro, S., Mecca, T., Seghezzi, M., Manenti, B., Azzarà, G., Dominoni, P., Crippa, A., Ottomano, C., and Lippi, G.

Subjects
ERYTHROCYTES, AUTOMATION, BLOOD testing, BLOOD platelets, BIOELECTRIC impedance, LEUCOCYTES, MEDICAL technology, REFERENCE values, REGRESSION analysis, RETICULOCYTES, DATA analysis software, AUTOANALYZERS, PLATELET count
Abstract

Introduction This study was aimed to compare the analytical performance of traditional and new parameters and morphological flags of CAL-8000 and XN-9000. The automated differential leukocyte count ( DIFF profile) and morphological flags were compared with optical microscopy ( OM). Methods A total of 1025 peripheral blood samples, collected in K3 EDTA tubes, were analyzed by CAL-8000, by XN-9000, and by OM. Within-run imprecision was performed in low cellularity samples. The comparison was made using Spearman's correlation, Passing-Bablok regression, Bland-Altman bias, and Cohen's K test. Results Within-run imprecision in low cellularity samples yielded reproducible data between the instruments (imprecision was higher than 10% on samples with platelet count <21 × 109/L using impedance technology). Passing-Bablok regression ( CAL-8000 vs. XN-9000) yielded slopes ranging between 0.2 to 1.16 and intercepts from −6.54 to 21.63. The bias for leukocytes parameters ranged from −1.8% to −82.2%, the red blood cell parameters from −2.9% to 3.1%, platelets parameters from −27.8% to 26%, and reticulocyte parameters from −115.3% to 4.5%. The comparison of morphological flags yielded a K value always <0.55. The DIFF profile vs. OM had a Passing-Bablok regression with slopes ranging between 0.34 to 1.00 and intercepts from −0.01% to 0.11 and bias ranging from −42.9% to 2.6% for XN-9000 parameters and from −2.7% to 35.0% for CAL-8000 parameters. The comparison of morphological flags showed a K value ranging from 0.35 to 0.77 for XN-9000 and from 0.17 to 0.54 for CAL-8000. Conclusion Differences exist between the two analyzers, especially in the generation of morphology flags, thus emphasizing the need of pursuing a major degree of harmonization and/or adopting instrument-specific reference ranges. [ABSTRACT FROM AUTHOR]

Published
2017
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46. Evaluation of nucleated red blood cell count by Sysmex XE-2100 in patients with thalassaemia or sickle cell anaemia and in neonates

Author

Buoro, Sabrina, Vavassori, Mauro, Pipitone, Silvia, Benegiamo, Anna, Lochis, Eleonora, Fumagalli, Sabina, Falanga, Anna, Marchetti, Marina, Crippa, Alberto, Ottomano, Cosimo, Lippi, Giuseppe, Buoro, S, Vavassori, M, Pipitone, S, Benegiamo, A, Lochis, E, Fumagalli, S, Falanga, A, Marchetti, M, Crippa, A, Ottomano, C, and Lippi, G

Subjects
Adult, Male, analysers, Adolescent, Erythroblasts, nucleated red blood cell, analysers, haematology, Anemia, Sickle Cell, Sensitivity and Specificity, Young Adult, nucleated red blood cells, XE-2100, thalassaemia, sickle cell disease, neonates, Humans, False Positive Reactions, Fluorometry, Child, False Negative Reactions, Observer Variation, beta-Thalassemia, Infant, Newborn, Infant, Middle Aged, ROC Curve, Area Under Curve, Child, Preschool, haematology, Erythrocyte Count, nucleated red blood cell, Original Article, Female
Abstract

Background. Current haematology analysers have variable sensitivity and accuracy for counting nucleated red blood cells in samples with low values and in all those conditions characterised by altered sensitivity of red blood cells to the lysing process, such as in beta-thalassaemia or sickle-cell diseases and in neonates. The aim of our study was to evaluate the performance of the automated analyser XE-2100 at counting nucleated red blood cells in the above-mentioned three categories of subjects with potentially altered red blood cell lysis sensitivity and yet a need for accurate nucleated red blood cell counts. Materials and methods. We measured nucleated red blood cell count by XE-2100 in peripheral blood samples of 187 subjects comprising 55 patients with beta-thalassaemia (40 major and 15 traits), 26 sickle-cell patients, 56 neonates and 50 normal subject. Results were compared with those obtained by optical microscopy. Agreement between average values of the two methods was estimated by means of Pearson's correlation and bias analysis, whereas diagnostic accuracy was estimated by analysis of receiver operating characteristic curves. Results. The comparison between the two methods showed a Pearson's correlation of 0.99 (95% CI; 0.98-0.99; p

Published
2015

47. Renal and Metabolic Effects of Insulin Lispro in Type 2 Diabetic Subjects With Overt Nephropathy

Author

Roberto Trevisan, Piero Ruggenenti, Giuseppe Remuzzi, Claudio Aros, Claudio Flores, Bogdan Ene-Iordache, Cosimo Ottomano, Ruggenenti, P, Flores, C, Aros, C, Ene-Iordache, B, Trevisan, R, Ottomano, C, and Remuzzi, G

Subjects
Adult, Blood Glucose, Male, medicine.medical_specialty, Endocrinology, Diabetes and Metabolism, Insulin analog, Renal function, Kidney, Renal Circulation, Internal medicine, Internal Medicine, medicine, Albuminuria, Humans, Hypoglycemic Agents, Insulin, Insulin lispro, Diabetic Nephropathies, Aged, Advanced and Specialized Nursing, insulin analog lispro, glomerular hyperfiltration, tyoe 2 diabetes, macroalbuminuria, Cross-Over Studies, Insulin Lispro, business.industry, Middle Aged, Glucose clamp technique, Postprandial Period, Endocrinology, Postprandial, Renal blood flow, Multivariate Analysis, Glucose Clamp Technique, Regular insulin, Female, Vascular Resistance, business, Glomerular hyperfiltration, Glomerular Filtration Rate, medicine.drug
Abstract

OBJECTIVE—To assess whether the insulin analog lispro may antagonize the renal effects of IGF-1, a mediator of glomerular hyperfiltration involved in the progression of diabetic and nondiabetic chronic nephropathies. RESEARCH DESIGN AND METHODS—In a randomized crossover study, we compared the renal and metabolic responses to regular or lispro insulin (0.1 units/kg body wt) administered after a euglycemic clamp and 5 and 30 min before a standard meal to 11 type 2 diabetic patients with macroalbuminuria. RESULTS—Two- and four-hour postprandial changes (vs. preprandial euglycemia) in glomerular filtration rate (GFR) followed a significantly different trend (5.8 ± 5.0 vs. −6.3 ± 4.7, P < 0.05; and 11.0 ± 6.8 vs. 0.7 ± 5.1%, P < 0.05) after regular insulin and lispro injection, respectively. After lispro, postprandial GFR changes were negatively correlated (r = −0.48, P = 0.0001) with plasma insulin concentration. After regular insulin, renal plasma flow increased in parallel with a decrease in renal vascular resistances. Both changes were fully prevented by lispro. Postprandial blood glucose maximum concentration (278 ± 16 vs. 240 ± 16 mg/dl, P < 0.01) and area under the curve (79,381 ± 19,237 vs. 72,810 ± 16,211 mg/dl per min, P < 0,05) were significantly lower after insulin lispro than after regular insulin injection, respectively, despite comparable postprandial insulin profiles. Changes in total and gluconeogenic amino acids followed a similar trend. Changes in blood glucose and plasma amino acids did not correlate with concomitant changes in GFR. CONCLUSIONS—In overt nephropathy of type 2 diabetes, lispro prevents glomerular hyperfiltration and offsets the renal effects of meal or meal-associated hyperglycemia by mechanisms possibly related to IGF-1 antagonism.

Published
2003

48. Analytical evaluation of Sysmex UF-1000i for flow cytometric analysis of peritoneal fluid

Author

Maria Grazia Lucà, Sabrina Buoro, Paola Dominoni, Patrizia Filisetti, Cosimo Ottomano, Giuseppe Lippi, Rosanna Gustinetti, Stefano Fagiuoli, Maria Grazia Alessio, Buoro, S, Gustinetti, R, Dominoni, P, Alessio, M, Filisetti, P, Lucà, M, Fagiuoli, S, Ottomano, C, and Lippi, G

Subjects
Pathology, medicine.medical_specialty, Clinical Biochemistry, Cell Count, Analytical evaluation, flow cytometry, peritoneal fluid, Sensitivity and Specificity, Flow cytometry, Peritoneal fluid, Nucleated cell, White blood cell, medicine, Ascitic Fluid, Humans, Evaluation, Chromatography, medicine.diagnostic_test, business.industry, Analytical performance, Reproducibility of Results, General Medicine, Flow Cytometry, medicine.anatomical_structure, Regression Analysis, business
Abstract

Objective: To evaluate analytical performance of Sysmex UF-1000i for peritoneal fluid analysis. Methods: Functional sensitivity, imprecision, linearity and comparison studies were performed on peritoneal fluids. Results: Total imprecision was 1.6-4.7%, functional sensitivity 27/μL for white blood cell (WBC) and 32/μL for total nucleated cell (TNC) count. Linearity was excellent up to 983. cell/μL, carry-over < 0.2%, correlation with manual microscopy always greater than 0.992. Conclusion: The instrument exhibited optimal performance at the conventional WBC diagnostic thresholds.

Published
2012

49. IgA anti-actin antibodies ELISA in coeliac disease: A multicentre study

Author

Ignazio Brusca, Giuseppe Ambrosiano, Maria Barrale, Saverio Teresi, S. M. La Chiusa, Aurelio Sonzogni, L. Di Prima, Giuseppe Iacono, A. D’Angelo, C. Scalici, Giuseppe Pirrone, Antonio Carroccio, M. G. Alessio, B. Cefalù, C. Ottomano, Carroccio, A., Brusca, I., Iacono, G., Alessio, M., Sonzogni, A., DI PRIMA, L., Barrale, M., Ottomano, C., Ambrosiano, G., Teresi, S., D'Angelo, A., Pirrone, G., Cefalu', A., Scalici, C., and LA CHIUSA SM

Subjects
Adult, Male, Pathology, medicine.medical_specialty, Settore MED/09 - Medicina Interna, Adolescent, Enzyme-Linked Immunosorbent Assay, Serum iga, Disease, Commercial kit, Sensitivity and Specificity, Coeliac disease, IgA anti-actin antibodies, coeliac disease, multicentre study, Intestinal mucosa, Humans, Medicine, Intestinal Mucosa, Villous atrophy, Child, Aged, Autoantibodies, Hepatology, biology, business.industry, Gastroenterology, Infant, nutritional and metabolic diseases, Middle Aged, IgA anti-actin antibodie, medicine.disease, Actins, Immunoglobulin A, Celiac Disease, Intestinal histology, Child, Preschool, biology.protein, Female, Antibody, business, Biomarkers
Abstract

Previous studies have demonstrated that serum anti-actin antibodies are a reliable marker of intestinal damage severity in coeliac disease.To validate in a multicentre study the clinical usefulness of serum IgA anti-actin antibody ELISA and its possible use in monitoring intestinal mucosa lesions during gluten-free diet.Four centres recruited 205 newly diagnosed coeliac disease patients with villous atrophy, 80 healthy controls and 81 "disease" controls. Twelve coeliac disease patients on gluten-free diet but with persistent symptoms underwent serum IgA anti-actin antibody assay and intestinal histology evaluation. IgA anti-actin antibody ELISA was performed with a commercial kit. All coeliac disease patients underwent intestinal histology study.IgA anti-actin antibodies showed a sensitivity of 80% and a specificity of 85% in the diagnosis of coeliac disease patients with villous atrophy. The area under the receiving operator curve for anti-actin antibodies was 0.873 [95% C.I. 0.805-0.899]. Serum anti-actin antibodies values were significantly higher in coeliac disease patients than in healthy or "disease" controls (P0.0001). Serum anti-actin antibodies were positive in 41 of the 60 coeliac disease patients with mild intestinal histology lesions (69%) and in 123 of the 145 with severe lesions (85.3%) (P0.05). There was a significant inverse correlation between anti-actin antibody values and the villi/crypts ratio (r=-0.423; P0.0001). In the 12 coeliac disease patients on gluten-free diet who underwent re-evaluation as they were persistently symptomatic, intestinal histology showed three cases with persistent villous atrophy: all of these were positive for serum anti-actin antibodies ELISA, whereas both serum anti-tTG and EmAs were negative. The other nine patients showed normal intestinal villi and were negative for serum anti-actin antibodies.Anti-actin antibodies are a reliable marker of severe intestinal mucosa damage in coeliac disease patients and a simple ELISA technique offers an accurate method for their determination. These antibodies seem to be a very reliable marker of persistent intestinal damage in coeliac disease patients.

Published
2007

50. Evaluation of residual humoral immune response against SARS-CoV-2 by a surrogate virus neutralization test (sVNT) 9 months after BNT162b2 primary vaccination.

Author

Pezzati L, Milazzo L, Carrozzo G, Kullmann C, Oreni L, Beltrami M, Caronni S, Lai A, Caberlotto L, Ottomano C, Antinori S, and Ridolfo AL

Subjects
Humans, Immunity, Humoral, BNT162 Vaccine, COVID-19 Vaccines, Cross-Sectional Studies, Neutralization Tests, Antibodies, Neutralizing, Immunoglobulin G, Antibodies, Viral, Vaccination, SARS-CoV-2, COVID-19 prevention & control
Abstract

The humoral response to SARS-CoV-2 vaccination has shown to be temporary, although may be more prolonged in vaccinated individuals with a history of natural infection. We aimed to study the residual humoral response and the correlation between anti-Receptor Binding Domain (RBD) IgG levels and antibody neutralizing capacity in a population of health care workers (HCWs) after 9 months from COVID-19 vaccination. In this cross-sectional study, plasma samples were screened for anti-RBD IgG using a quantitative method. The neutralizing capacity for each sample was estimated by means of a surrogate virus neutralizing test (sVNT) and results expressed as the percentage of inhibition (%IH) of the interaction between RBD and the angiotensin-converting enzyme. Samples of 274 HCWs (227 SARS-CoV-2 naïve and 47 SARS-CoV-2 experienced) were tested. The median level of anti-RBD IgG was significantly higher in SARS-CoV-2 experienced than in naïve HCWs: 2673.2 AU/mL versus 610.9 AU/mL, respectively (p <0.001). Samples of SARS-CoV-2 experienced subjects also showed higher neutralizing capacity as compared to naïve subjects: median %IH = 81.20% versus 38.55%, respectively; p <0.001. A quantitative correlation between anti-RBD Ab and inhibition activity levels was observed (Spearman's rho = 0.89, p <0.001): the optimal cut-off correlating with high neutralization was estimated to be 1236.1 AU/mL (sensitivity 96.8%, specificity 91.9%; AUC 0.979). Anti-SARS-CoV-2 hybrid immunity elicited by a combination of vaccination and infection confers higher anti-RBD IgG levels and higher neutralizing capacity than vaccination alone, likely providing better protection against COVID-19., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Japanese Society of Chemotherapy, Japanese Association for Infectious Diseases, and Japanese Society for Infection Prevention and Control. Published by Elsevier Ltd. All rights reserved.)

Published
2023
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