75 results on '"Salvianti F"'
Search Results
2. Microbial calcium carbonate precipitation for reinforcement of monumental stones
- Author
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Barabesi, C., primary, Salvianti, F., additional, Mastromei, G., additional, and Perito, B., additional
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- 2017
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3. Early Detection of Fungal Plant Pathogens by Real-Time Quantitative PCR: The Case of Diplodia sapinea on Pine
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Luchi N., Santini A., Salvianti F., and Pinzani P.
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fungi ,food and beverages ,Fungal pathogens Latent infections qPCR Diplodia shoot blight Conifers - Abstract
This chapter reports the use of real-time quantitative PCR to detect Diplodia sapinea, a fungal plant pathogen that causes shoot tip dieback and tree mortality on pine trees. This molecular approach represents a reliable and sensitive tool to detect fungal pathogens in DNA extracted from plant tissues and its use can be also recommended to study fungal behavior in host tissues by quantifying fungal growth in the latent phase, when symptoms in the host are not present yet.
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- 2020
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4. Circulating BRAFV600E in the Diagnosis and Follow-Up of Differentiated Papillary Thyroid Carcinoma
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Pupilli, C., Pinzani, P., Salvianti, F., Fibbi, B., Rossi, M., Petrone, L., Perigli, G., De Feo, M. L., Vezzosi, V., Pazzagli, M., Orlando, C., and Forti, G.
- Published
- 2013
5. miR-20a-5p expression as a potential non-invasive diagnostic biomarker in patients with non-obstructive azoospermia
- Author
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Cito, G., primary, Pinzani, P., additional, Fucci, R., additional, Picone, R., additional, Salvianti, F., additional, Giachini, C., additional, Falcone, P., additional, Micelli, E., additional, Cocci, A., additional, Verrienti, P., additional, Minervini, A., additional, Carini, M., additional, Coccia, M.E., additional, and Natali, A., additional
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- 2020
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6. 671P Prospective assessment of AR splice variant and PSMA detection on circulating tumour cells of mCRPC patients: Preliminary results of PRIMERA trial (NCT04188275)
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Francolini, G., primary, Salvestrini, V., additional, Loi, M., additional, Mangoni, M., additional, Detti, B., additional, Pinzani, P., additional, Salvianti, F., additional, Desideri, I., additional, Aquilano, M., additional, Mariotti, M., additional, Garlatti, P., additional, Stocchi, G., additional, Ciccone, L.P., additional, Lucidi, S., additional, Salvatore, G., additional, Sottili, M., additional, Meattini, I., additional, and Livi, L., additional
- Published
- 2020
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7. Corrigendum to “The pre-analytical phase of liquid biopsy” [New Biotechnol. 55, 25 March 2020, 19–29]
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Salvianti, F., primary, Gelmini, S., additional, Costanza, F., additional, Mancini, I., additional, Sonnati, G., additional, Simi, L., additional, Pazzagli, M., additional, and Pinzani, P., additional
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- 2020
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8. Evidenza di cellule tumorali circolanti in pazienti con carcinoma surrenalico: studio preliminare monocentrico
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Pinzani, P., Scatena, C., Salvianti, F., Corsini, E., Canu, L., Poli, G., Paglierani, M., Piccini, V., Pazzagli, M., Nesi, G., Mannelli, M., Luconi, M., and Arvat, Emanuela
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- 2013
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9. SC10 - miR-20a-5p expression as a potential non-invasive diagnostic biomarker in patients with non-obstructive azoospermia
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Cito, G., Pinzani, P., Fucci, R., Picone, R., Salvianti, F., Giachini, C., Falcone, P., Micelli, E., Cocci, A., Verrienti, P., Minervini, A., Carini, M., Coccia, M.E., and Natali, A.
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- 2020
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10. CirculatingBRAFV600Ein the Diagnosis and Follow-Up of Differentiated Papillary Thyroid Carcinoma
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Pupilli, C., primary, Pinzani, P., additional, Salvianti, F., additional, Fibbi, B., additional, Rossi, M., additional, Petrone, L., additional, Perigli, G., additional, De Feo, M. L., additional, Vezzosi, V., additional, Pazzagli, M., additional, Orlando, C., additional, and Forti, G., additional
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- 2013
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11. Molecular Analysis of Single Circulating Tumor Cells (CTCS) Isolated from Metastatic Breast Cancer (MBC) Patients (PTS)
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Pestrin, M., primary, Galardi, F., additional, Salvianti, F., additional, De Luca, F., additional, Bessi, S., additional, Capaccioli, G., additional, Di Leo, A., additional, Giannini, A., additional, Pinzani, P., additional, and Pazzagli, M., additional
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- 2013
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12. Transplantation: basic science
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Cantaluppi, V., primary, De Lena, M., additional, Beltramo, S., additional, Ferrario, S., additional, Dellepiane, S., additional, Figliolini, F., additional, Bruno, S., additional, Biancone, L., additional, Segoloni, G. P., additional, Tetta, C., additional, Camussi, G., additional, Prasad, N., additional, Jaisawal, A., additional, Yadav, B., additional, Agarwal, V., additional, Tripathi, D., additional, Nunez-Lozano, R., additional, Quiros, Y., additional, Sanchez-Gonzalez, P., additional, Perez de Obanos, M. P., additional, Ruiz, J., additional, Lopez-Hernandez, F. J., additional, Lopez-Novoa, J. M., additional, Yang, J. W., additional, Kim, J. S., additional, Lee, J. Y., additional, Park, H. C., additional, Han, B. G., additional, Choi, S. O., additional, Matsuyama, M., additional, Yoshimura, R., additional, Hayama, T., additional, Chargui, J., additional, Touraine, J.-L., additional, Yoshimura, N., additional, Zanazzi, M., additional, Carta, P., additional, Caroti, L., additional, Antognoli, G., additional, Pinzani, P., additional, Salvianti, F., additional, Villari, D., additional, Minetti, E., additional, Genina, A., additional, Ismail, W., additional, Soliman, A., additional, Ucar, H., additional, Akbas, H. S., additional, Yilmaz, V. T., additional, Aktas, A., additional, Suleymanlar, G., additional, Yucel, G., additional, Cappuccilli, M. L., additional, La Manna, G., additional, Capelli, I., additional, Baraldi, O., additional, Cuna, V., additional, Battaglino, G., additional, Todeschini, P., additional, Feliciangeli, G., additional, Scolari, M. P., additional, Stefoni, S., additional, Loiacono, E., additional, Votta, B., additional, Amore, A., additional, Ranghino, A., additional, Camilla, R., additional, Peruzzi, L., additional, Donadio, M. E., additional, Serriello, I., additional, Gallo, R., additional, Puccinelli, M. P., additional, Coppo, R., additional, Sahin, G., additional, Meltem Akay, O., additional, Uslu, S., additional, Bal, C., additional, Ugur Yalcin, A., additional, Gulbas, Z., additional, and George, J., additional
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- 2013
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13. Atypical Spitzoid melanocytic tumors: A morphological, mutational, and FISH analysis.
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Massi D, Cesinaro AM, Tomasini C, Paglierani M, Bettelli S, Dal Maso L, Simi L, Salvianti F, Pinzani P, Orlando C, De Giorgi V, Lukic S, Maiorana A, Santucci M, and Canzonieri V
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- 2011
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14. Prospective evaluation of RASSF1A cell-free DNA as a biomarker of pre-eclampsia
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Laura Cremonesi, Luca Valsecchi, Francesca Salvianti, Mario Pazzagli, Silvia Galbiati, Maddalena Smid, Annalisa Inversetti, Massimo Candiani, Maurizio Ferrari, Pamela Pinzani, Salvianti, F, Inversetti, A, Smid, M, Valsecchi, L, Candiani, Massimo, Pazzagli, M, Cremonesi, L, Ferrari, Maurizio, Pinzani, P, and Galbiati, S.
- Subjects
Adult ,medicine.medical_specialty ,RASSF1A cell-free DNA ,Preeclampsia ,Pre-Eclampsia ,Predictive Value of Tests ,Pregnancy ,medicine ,Humans ,Promoter Regions, Genetic ,Fetus ,Eclampsia ,Predictive marker ,business.industry ,Obstetrics ,Tumor Suppressor Proteins ,Obstetrics and Gynecology ,Gestational age ,Biomarker, Maternal plasma ,DNA ,DNA Methylation ,medicine.disease ,Epidemiologic Studies ,Reproductive Medicine ,Cell-free fetal DNA ,Immunology ,Gestation ,Biomarker (medicine) ,Female ,business ,Biomarkers ,Developmental Biology - Abstract
Introduction This study aims to quantify total and fetal cell-free DNA (cfDNA) in maternal plasma at different gestational ages and to assess whether this could represent a reliable predictive marker of pre-eclampsia (PE) before clinical onset. Methods We performed a qPCR assay to compare the cfDNA concentration of hypermethylated and unmethylated RASSF1A promoter gene sequences in maternal plasma among 3 groups of pregnant women. These included 17 women with overt PE, 33 women at risk for the disease subsequently differentiated into 9 who developed PE and 24 who did not, and 73 controls. All women at risk were consecutively sampled throughout the whole gestation. Results Both total and fetal cfDNA had a good diagnostic performance in distinguishing patients with overt PE from healthy controls. When comparing women at risk who developed PE to women at risk who did not, the predictive capability was satisfactory at a gestational age ranging from 17 to 30 weeks. This allowed establishing within this time interval a cut-off value of 735 GE/ml for total cfDNA (87.5% sensitivity and 70.0% specificity), and a cut-off value of 7.49 GE/ml for fetal cfDNA (100% sensitivity and 50% specificity). cfDNA levels turned positive several weeks before the onset of the disease: from 2 to 18 weeks for total cfDNA and from 8 to 17 weeks for fetal cfDNA. Discussion The simultaneous use of total and fetal cfDNA would allow an accurate monitoring and prevention of PE development thus suggesting that RASSF1A could represent a potential biomarker of PE.
- Published
- 2015
15. Atypical Spitz tumors in patients younger than 18 years
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Angela Rita Sementa, Claudio Gambini, Paola Collini, Maria Elena Errico, Milena Paglierani, Francesca Salvianti, Rebecca Senetta, Gabrina Tragni, Carlo Tomasini, Franco Rongioletti, Vittoria Donofrio, Maria Cristina Montesco, Andrea Ferrari, Daniela Massi, Renata Boldrini, Massi, D, Tomasini, C, Senetta, R, Paglierani, M, Salvianti, F, Errico, Me, Donofrio, V, Collini, P, Tragni, G, Sementa, Ar, Rongioletti, F, Boldrini, R, Ferrari, A, Gambini, C, and Montesco, Mc
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Male ,medicine.medical_specialty ,Pathology ,Skin Neoplasms ,Adolescent ,medicine.medical_treatment ,skin neoplasms ,Sentinel lymph node ,Dermatology ,Epithelioid and Spindle Cell ,preschool ,male ,nevus epithelioid and spindle cell ,Nevus, Epithelioid and Spindle Cell ,Medicine ,Humans ,In patient ,humans ,Child ,Preschool ,fluorescence in situ hybridization ,Nevus ,Retrospective Studies ,child ,medicine.diagnostic_test ,business.industry ,Melanoma ,atypical Spitz tumor ,pediatric age ,sentinel lymph nodevSpitz nevus ,Child, Preschool ,Female ,Infant ,Retrospective cohort study ,Pediatric age ,medicine.disease ,Spitz nevus ,infant ,retrospective studies ,female ,adolescent ,Lymphadenectomy ,business ,Fluorescence in situ hybridization - Abstract
Background Diagnosis and proper management of atypical Spitz tumors in pediatric age are still controversial. Objective We sought to investigate the clinicopathological and molecular features of atypical Spitz tumors in patients aged 18 years or younger. Methods We performed a retrospective clinicopathological and fluorescence in situ hybridization study on 50 pediatric atypical Spitz tumors. Results Parameters that were significantly correlated with a diagnosis of atypical Spitz tumors over Spitz nevus included asymmetry, level IV/V, lack of maturation, solid growth, nuclear pleomorphism, high nuclear-cytoplasmic ratio, atypical and deep mitoses, and more than 6 mitoses/mm 2 . In the atypical Spitz tumors group, a significantly higher mitotic rate was observed in prepuberal age ( P = .04). The 4-probe fluorescence in situ hybridization melanoma assay did not discriminate atypical Spitz tumors from Spitz nevi. Heterozygous 9p21 loss was found in 3 of 37 cases and homozygous 9p21 loss in 2 of 37 cases. Only 1 child experienced a fatal outcome, showing genetic abnormalities by melanoma fluorescence in situ hybridization probe and a heterozygous 9p21 deletion. Limitations The limited number of adverse outcomes did not allow the prognostic analysis of single morphologic features. Conclusion Pediatric atypical Spitz tumors are associated with minimal lethal potential. Atypical Spitz tumors require complete excision and careful follow-up while our data do not support any clinical benefit for the sentinel lymph node biopsy procedure and completion lymphadenectomy.
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- 2015
16. Atypical Spitzoid melanocytic tumors:A morphological, mutational, and FISH analysis
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Antonio Maiorana, Milena Paglierani, Silvana Lukic, Marco Santucci, Vincenzo Canzonieri, Daniela Massi, Francesca Salvianti, Luigino Dal Maso, Carlo Tomasini, Anna Maria Cesinaro, Vincenzo De Giorgi, Claudio Orlando, Lisa Simi, Pamela Pinzani, Stefania Bettelli, Massi, D, Cesinaro, Am, Tomasini, C, Paglierani, M, Bettelli, S, Dal Maso, L, Simi, L, Salvianti, F, Pinzani, P, Orlando, C, De Giorgi, V, Lukic, S, Maiorana, A, Santucci, M, and Canzonieri, V
- Subjects
Adult ,Male ,Proto-Oncogene Proteins B-raf ,Pathology ,medicine.medical_specialty ,Skin Neoplasms ,atypical Spitz nevus, atypical Spitzoid tumor, BRAFV600E, fluorescence in situ hybridization ,Adolescent ,Sentinel lymph node ,Lymph node biopsy ,Dermoscopy ,Dermatology ,Gene mutation ,Ether ,Young Adult ,BRAFV600E ,Formaldehyde ,Nevus, Epithelioid and Spindle Cell ,Biopsy ,medicine ,Humans ,Child ,Lymph node ,fluorescence in situ hybridization ,In Situ Hybridization, Fluorescence ,Acetic Acid ,Chromatography ,Ethanol ,medicine.diagnostic_test ,business.industry ,Micrometastasis ,Infant ,atypical Spitzoid tumor ,Middle Aged ,atypical Spitz nevus ,Prognosis ,Immunohistochemistry ,Genes, ras ,medicine.anatomical_structure ,Child, Preschool ,Female ,business ,Fluorescence in situ hybridization ,Comparative genomic hybridization - Abstract
Background: Identification of the clinical behavior of atypical Spitzoid tumors with conflicting histopathologic features remains controversial. Objective: We sought to assess whether molecular findings may be helpful in the diagnostic and prognostic assessment of atypical Spitzoid tumors. Methods: A total of 38 controversial, atypical Spitzoid lesions (1 mm in thickness) were analyzed for clinicopathological features, chromosomal alterations by fluorescence in situ hybridization (FISH) analysis (RREB1/MYB/CCND1/CEP6), BRAF(V600E) mutation by allele-specific real-time polymerase chain reaction confirmed by sequencing, and H-RAS gene mutation by direct sequencing. Results: Atypical Spitzoid lesions developed in 21 female and 17 male patients (mean age 22 years). Nine patients underwent sentinel lymph node biopsy and a sentinel lymph node micrometastasis was detected in 4 of these 9 cases. Four additional patients, who did not receive a sentinel lymph node biopsy, experienced bulky lymph node metastases and one experienced visceral metastases and death. Lesions from patients with lymph node involvement showed more deep mitoses (P < .01), less inflammation = .05), and more plasma cells (P = .04). FISH analysis demonstrated the presence of chromosomal alterations in 6 of 25 cases. Correlation with follow-up data showed that the only case with fatal outcome showed multiple chromosomal alterations by FISH analysis. BRAF(V600E) mutation was detected in 12 of 16 cases (75%) and H-RAS mutation on exon 3 was found in 3 of 11 cases (27%). Limitations: Our results require validation in a larger series with longer follow-up information. Conclusions: FISH assay may be of help in the prognostic evaluation of atypical Spitzoid tumors. Diagnostic significance of BRAF(V600E) and H-RAS mutations in this setting remains unclear. (J Am Acad Dermatol 2011;64:919-35)
- Published
- 2011
17. Early changes in circulating tumor DNA (ctDNA) predict treatment response in metastatic KRAS-mutated colorectal cancer (mCRC) patients.
- Author
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Lavacchi D, Gelmini S, Calabri A, Rossi G, Simi L, Caliman E, Mancini I, Salvianti F, Petroni G, Guidolin A, Scolari F, Messerini L, Pillozzi S, Pinzani P, and Antonuzzo L
- Abstract
The detection of RAS mutations and co-mutations in liquid biopsy offers a novel paradigm for the dynamic management of metastatic colorectal cancer (mCRC) patients. Expanding the results of the prospective OMITERC (OMIcs application from solid to liquid biopsy for a personalized ThERapy of Cancer) project, we collected blood samples at specific time points from patients who received a first-line chemotherapy (CT) for KRAS-mutated mCRC. CTC quantification was performed by CellSearch® system. Libraries from cfDNA were prepared using the Oncomine™ Colon cfDNA Assay to detect tumour-derived DNA in cfDNA. The analysis involved >240 hotspots in 14 genes. Twenty patients with KRAS-mutated mCRC treated at the Medical Oncology Unit of Careggi University Hospital were prospectively enrolled. Nine patients had available data for longitudinal monitoring of cfDNA. After 6 weeks of first-line CT an increase of KRAS-mutated clone was reported in the only patient who did not obtain disease control, while all patients with decrease of KRAS clones obtained disease control. Overall, in patients with a short (<9 months) progression-free survival (PFS) we registered, at 6 weeks, an increase in cfDNA levels and in KRAS mutations or other co-mutations, i.e. PIK3CA, FBXW7, GNAS, and TP53. In selected cases, co-mutations were able to better anticipate radiological progressive disease (PD) than the increase of KRAS-mutated clones. In conclusion, our study confirms plasma ctDNA as a crucial tool for anticipating PD at an early time point and highlights the value of a comprehensive assessment of clonal dynamics to improve the management of patients with mCRC., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Lorenzo Antonuzzo reports financial support was provided by 10.13039/501100009888Tuscany Region. Lorenzo Anronuzzo reports a relationship with 10.13039/501100009888Tuscany Region that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)
- Published
- 2023
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18. Safe and Successful Surgical Outcome in Persons with Hemophilia A with and without Inhibitors Treated with Emicizumab: A Large, Single Center, Real-World Experience.
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Castaman G, Linari S, Pieri L, Carulli C, Prosperi P, Tonelli P, Demartis F, Fjerza R, Attanasio M, Coppo M, and Salvianti F
- Abstract
Emicizumab is a humanized recombinant bispecific antibody, bridging together activated factor IX (FIXa) and factor X (FX), thus mimicking the activity of FVIII in vivo. Emicizumab is designed for long-term prophylaxis in patients with severe hemophilia A with and without inhibitors. This approach provides constant protection, with significant reduction in bleeding rate and improved quality of life. However, protection provided by emicizumab is not absolute, and clotting factor concentrates (FVIII, rFVIIa, aPCC) may be necessary for post-traumatic bleeding or surgery, with a potential thrombotic risk or difficulty in preventing bleeding. Real world evidence is still scanty, especially for managing major surgery. In this study, 75 surgeries were managed in 28 patients (27 major procedures in 15 patients and 48 minor procedures in 20 patients. In 17 patients without inhibitors, 30 minor surgeries were carried out by using FVIII in 5, with only a bleeding event, which was successfully treated with FVIII concentrate. Six major surgeries were uneventfully performed with FVIII concentrate. Eleven PWHA and high-titer inhibitors underwent 39 surgical procedures (18 minor and 21 major surgeries). Minor surgeries were mostly performed without prophylaxis with rFVIIa, with only a single bleeding complication. All 21 major surgeries were covered with a homogeneous protocol using rFVIIa. In four instances, bleeding complications occurred, treated with rFVIIa. Of them, a single patient only failed to respond and died because of an uncontrollable bleeding from a large ruptured retroperitoneal pseudotumor. Surgery in patients with emicizumab can be safely carried out with the use of appropriate replacement therapy protocols.
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- 2023
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19. Prospective assessment of AR splice variant and multi-biomarker expression on circulating tumor cells of mCRPC patients undergoing androgen receptor targeted agents: interim analysis of PRIMERA trial (NCT04188275).
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Francolini G, Loi M, Ciccone LP, Detti B, Di Cataldo V, Pinzani P, Salvianti F, Salvatore G, Sottili M, Santini C, Frosini G, Visani L, Burchini L, Mattioli C, Allegra AG, Valzano M, Cerbai C, Aquilano M, Salvestrini V, Desideri I, Mangoni M, Meattini I, and Livi L
- Subjects
- Biomarkers, Tumor genetics, Humans, Male, Prospective Studies, Prostate-Specific Antigen, Receptors, Androgen genetics, Receptors, Androgen metabolism, Treatment Outcome, Antineoplastic Agents therapeutic use, Neoplastic Cells, Circulating pathology, Prostatic Neoplasms, Castration-Resistant drug therapy, Prostatic Neoplasms, Castration-Resistant genetics
- Abstract
Circulating tumor cells detection and ARV7 expression are associated with worse clinical outcomes in metastatic Castration-Resistant Prostate Cancer (mCRPC) undergoing Androgen Receptor Targeted Agents. ARFL, PSMA and PSA may help to refine prognostic models. In our institution, a prospective observational trial testing CTC detection in mCPRC undergoing I line ARTA therapy terminated the planned enrollment in 2020. Here, we present a pre-planned interim analysis with 18 months of median follow-up. RT-qPCR was used to determine the CTC expression of PSA, PSMA, AR and ARV7 before starting ARTA. PSA-drop, Progression-Free and Overall Survival (PFS and OS) and their correlation with CTC detection were reported. Forty-four patients were included. CTC were detected in 43.2% of patients, of whom 8.94% expressed PSA, 15.78% showed ARV7, 63.15% and 73.68% displayed ARFL and PSMA, respectively. Biochemical response was significantly improved in CTC + vs CTC- patients, with median PSA-drop of 18.5 vs 2.5 ng/ml (p = 0.03). After a median follow-up of 18 months, 50% of patients progressed. PFS was significantly longer in CTC- patients (NR vs 16 months). Eight (18.2%) patients died, a non-significant trend in terms of OS was detected in favor of CTC- patients (NR vs 29 months, p = 0.05). AR, PSA and PSMA expression in CTC + had no significant impact on PSA-drop, PFS or OS. PRIMERA-trial confirmed the CTC detection predictive importance in mCRPC patients., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
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- 2022
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20. Circulating tumour cells and cell-free DNA as a prognostic factor in metastatic colorectal cancer: the OMITERC prospective study.
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Salvianti F, Gelmini S, Mancini I, Pazzagli M, Pillozzi S, Giommoni E, Brugia M, Di Costanzo F, Galardi F, De Luca F, Castiglione F, Messerini L, Pinzani P, and Antonuzzo L
- Subjects
- Aged, Aged, 80 and over, Colorectal Neoplasms genetics, Disease Progression, Female, High-Throughput Nucleotide Sequencing, Humans, Longitudinal Studies, Male, Middle Aged, Mutation, Neoplasm Metastasis, Prognosis, Prospective Studies, Biomarkers, Tumor genetics, Cell-Free Nucleic Acids genetics, Colorectal Neoplasms pathology, Neoplastic Cells, Circulating pathology, Proto-Oncogene Proteins p21(ras) genetics, Sequence Analysis, DNA methods
- Abstract
Background: Within the OMITERC prospective study (OMIcs application from solid to liquid biopsy for a personalised ThERapy of Cancer), we explored the prognostic role of liquid biopsy encompassing cell-free DNA (cfDNA) and circulating tumour cells (CTCs) in KRAS mutated metastatic colorectal cancer (mCRC)., Methods: We defined a workflow including pre-analytical and analytical procedures collecting blood before therapy and every 3 months until disease progression (PD). CTCs were counted by CellSearch® and isolated by DEPArray™. NGS sequencing of CTCs and cfDNA was performed using a panel of cancer/CRC related genes respectively., Results: KRAS mutational status was mostly concordant between tumour tissues and liquid biopsy. The percentage of cfDNA samples with mutations in CRC driver genes was in line with literature. In longitudinal monitoring circulating biomarkers anticipated or overlapped conventional diagnostic tools in predicting PD. The presence of CTCs at baseline was confirmed a negative prognostic marker., Conclusions: Cell-free DNA and CTCs are readily available candidates for clinical application in mCRC. While CTCs demonstrated a prognostic significance at baseline, cfDNA was confirmed an easily accessible material for monitoring the mutational status of the tumour over time. Moreover, in the longitudinal study, the two markers emerged as complementary in assessing disease progression.
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- 2021
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21. Updates on liquid biopsy: current trends and future perspectives for clinical application in solid tumors.
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Pinzani P, D'Argenio V, Del Re M, Pellegrini C, Cucchiara F, Salvianti F, and Galbiati S
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- Biomarkers, Tumor, Humans, Liquid Biopsy, Circulating Tumor DNA, MicroRNAs genetics, Neoplastic Cells, Circulating
- Abstract
Despite advances in screening and therapeutics cancer continues to be one of the major causes of morbidity and mortality worldwide. The molecular profile of tumor is routinely assessed by surgical or bioptic samples, however, genotyping of tissue has inherent limitations: it represents a single snapshot in time and it is subjected to spatial selection bias owing to tumor heterogeneity. Liquid biopsy has emerged as a novel, non-invasive opportunity of detecting and monitoring cancer in several body fluids instead of tumor tissue. Circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), RNA (mRNA and microRNA), microvesicles, including exosomes and tumor "educated platelets" were recently identified as a source of genomic information in cancer patients which could reflect all subclones present in primary and metastatic lesions allowing sequential monitoring of disease evolution. In this review, we summarize the currently available information concerning liquid biopsy in breast cancer, colon cancer, lung cancer and melanoma. These promising issues still need to be standardized and harmonized across laboratories, before fully adopting liquid biopsy approaches into clinical practice., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)
- Published
- 2021
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22. Prognostic and Monitoring Value of Circulating Tumor Cells in Adrenocortical Carcinoma: A Preliminary Monocentric Study.
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Cantini G, Canu L, Armignacco R, Salvianti F, De Filpo G, Ercolino T, Nesi G, Maggi M, Mannelli M, Pinzani P, and Luconi M
- Abstract
Adrenocortical carcinoma (ACC), a rare and aggressive neoplasia, presents poor prognosis when metastatic at diagnosis and limited therapies are available. Specific and sensitive markers for early diagnosis and a monitoring system of therapy and tumor evolution are urgently needed. The liquid biopsy represents a source of tumor material within a minimally invasive blood draw that allows the recovery of circulating tumor cells (CTCs). CTCs have been recently shown to be detectable in ACC. In the present paper, we evaluated the prognostic value of CTCs obtained by size-filtration in a small pilot cohort of 19 ACC patients. We found CTCs in 68% of pre-surgery and in 38% of post-surgery blood samples. In addition, CTC clusters (CTMs) and cancer associated macrophages (CAMLs) were detectable in some ACC patients. The median number of CTCs significantly decreased after the mass removal. Finally, stratifying patients in high and low pre-surgery CTC number groups, assuming the 75th percentile CTC value as cut-off, CTCs significantly predicted patients' overall survival (log rank = 0.005), also in a multivariate analysis adjusted for age and tumor stage. In conclusion, though preliminary and performed in a small cohort of patients, our study suggests that CTC number may represent a promising marker for prognosis and disease monitoring in ACC.
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- 2020
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23. Blood plasma miR-20a-5p expression as a potential non-invasive diagnostic biomarker of male infertility: A pilot study.
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Cito G, Coccia ME, Salvianti F, Fucci R, Picone R, Giachini C, Cocci A, Falcone P, Micelli E, Verrienti P, Minervini A, Carini M, Pinzani P, and Natali A
- Subjects
- Adult, Azoospermia pathology, Female, Humans, Pilot Projects, Azoospermia blood, Azoospermia diagnosis, Biomarkers blood, MicroRNAs blood
- Abstract
Background: Recently, alterations in miRNAs expression profile in semen have been linked to damaged spermatogenesis, suggesting miRNAs could be used as potential infertility biomarkers. In previous animal studies, miR-20a-5p was found to be down-expressed in low motile spermatozoa, implying its potential target of genes associated with cell apoptosis., Objective: To investigate miR-20a-5p expression in blood plasma of patients suffering from non-obstructive azoospermia (NOA), compared to normozoospermic controls., Materials and Methods: Between January 2018 and December 2019, from 52 infertile couples eligible for the study, 24 couples were finally enrolled in this monocentric observational prospective pilot study. Patients were included into two groups: Group 1 comprised men with NOA (n = 14) and Group 2 fertile men partners of women with female tubal factor infertility (n = 10). All NOA patients underwent testicular sperm extraction. The expression of circulating miR-20a-5p in plasma samples was assessed by RT-qPCR. A relative quantification strategy was adopted using the 2
-ΔCq method to calculate the target miR-20a-5p expression with respect to miR-16-5p as endogenous control., Results: Median blood plasma miR-20a-5p was significantly higher in patients affected by NOA (0.16 2-ΔCt , range: 0.05-0.79 2-ΔCt ) than in fertile controls (0.06 2-ΔCt , range: 0.04-0.10 2-ΔCt ), P < .001. MiR-20a-5p was positively correlated with follicle-stimulating hormone (FSH) (rrho = -0.490, P = .015) and luteinizing hormone (LH) (rrho = -0.462, P = .023), and negatively correlated with serum total testosterone (TT) (rrho = -0.534, P = .007) and right and left testicular size (rrho = -0.473, P = .020 and rrho = -0.471, P = .020, respectively). Successful sperm retrieval (SR) rate was 50.0%. Median value of miR-20a-5p did not differ significantly among patients with successful SR and those with negative SR. Testicular histological examination showed: hypospermatogenesis in 6/14 (42.8%), maturation arrest in 4/14 (28.6%), sertoli cell-only syndrome in 4/14 (28.6%). No significant differences in miR-20a-5p were found between histopathological patterns (P > .05)., Conclusions: MiR-20a-5p could represent a novel non-invasive diagnostic biomarker of male infertility., (© 2020 American Society of Andrology and European Academy of Andrology.)- Published
- 2020
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24. The pre-analytical phase of the liquid biopsy.
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Salvianti F, Gelmini S, Costanza F, Mancini I, Sonnati G, Simi L, Pazzagli M, and Pinzani P
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- Animals, Body Fluids metabolism, Cell-Free Nucleic Acids analysis, Exosomes metabolism, Humans, Neoplastic Cells, Circulating pathology, Liquid Biopsy methods, Pre-Analytical Phase methods
- Abstract
The term 'liquid biopsy', introduced in 2013 in reference to the analysis of circulating tumour cells (CTCs) in cancer patients, was extended to cell-free nucleic acids (cfNAs) circulating in blood and other body fluids. CTCs and cfNAs are now considered diagnostic and prognostic markers, used as surrogate materials for the molecular characterisation of solid tumours, in particular for research on tumour-specific or actionable somatic mutations. Molecular characterisation of cfNAs and CTCs (especially at the single cell level) is technically challenging, requiring highly sensitive and specific methods and/or multi-step processes. The analysis of the liquid biopsy relies on a plethora of methods whose standardisation cannot be accomplished without disclosing criticisms related to the pre-analytical phase. Thus, pre-analytical factors potentially influencing downstream cellular and molecular analyses must be considered in order to translate the liquid biopsy approach into clinical practice. The present review summarises the most recent reports in this field, discussing the main pre-analytical aspects related to CTCs, cfNAs and exosomes in blood samples for liquid biopsy analysis. A short discussion on non-blood liquid biopsy samples is also included., (Copyright © 2019 Elsevier B.V. All rights reserved.)
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- 2020
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25. Detection and Characterization of Circulating Tumor Cells by Quantitative Real-Time PCR.
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Salvianti F, Costanza F, Sonnati G, and Pinzani P
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- Biomarkers, Tumor genetics, DNA Mutational Analysis, Humans, Liquid Biopsy methods, Melanoma blood, Melanoma genetics, Melanoma pathology, Monophenol Monooxygenase genetics, RNA, Messenger genetics, Biomarkers, Tumor isolation & purification, Melanoma diagnosis, Neoplastic Cells, Circulating pathology, RNA, Messenger isolation & purification, Real-Time Polymerase Chain Reaction methods
- Abstract
We propose two different approaches involving the use of quantitative real-time PCR for the detection or analysis of circulating tumor cells. In one case cells are indirectly identified through the expression of a marker mRNA, while in the other one cells are enriched by size prior to be submitted to mutational analysis for a specific target. Both methods have been successfully applied to the study of circulating melanoma cells.
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- 2020
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26. Early Detection of Fungal Plant Pathogens by Real-Time Quantitative PCR: The Case of Diplodia sapinea on Pine.
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Luchi N, Santini A, Salvianti F, and Pinzani P
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- Ascomycota genetics, DNA, Fungal isolation & purification, Plant Diseases microbiology, Reproducibility of Results, Sensitivity and Specificity, Trees, Ascomycota isolation & purification, Pinus microbiology, Plant Diseases prevention & control, Real-Time Polymerase Chain Reaction methods
- Abstract
This chapter reports the use of real-time quantitative PCR to detect Diplodia sapinea, a fungal plant pathogen that causes shoot tip dieback and tree mortality on pine trees. This molecular approach represents a reliable and sensitive tool to detect fungal pathogens in DNA extracted from plant tissues and its use can be also recommended to study fungal behavior in host tissues by quantifying fungal growth in the latent phase, when symptoms in the host are not present yet.
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- 2020
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27. Quantitative Polymerase Chain Reaction Detection of Microchimerism in Female Transplant Renal Recipients.
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Villari D, Salvianti F, Zanazzi M, Martini A, Spatafora P, Caroassai Grisanti S, Sebastianelli A, Nicita G, Serni S, and Pinzani P
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- Adult, Aged, Cadaver, Female, Humans, Male, Middle Aged, Prospective Studies, Time Factors, Treatment Outcome, Chimerism, Kidney Transplantation, Polymerase Chain Reaction
- Abstract
Introduction: Microchimerism (MC) is the presence of a small amount of foreign cells or DNA within a person's circulation or tissues. It has been identified also in recipients of solid organ transplants where it seems to be critical for the development and maintenance of immunological tolerance. Nevertheless, natural and/or iatrogenic MC can be acquired prior to transplantation, through pregnancy and/or blood transfusion., Objective: The aim of this study was to detect the presence of MC in women after renal transplantation from male cadaveric donors and its relationship with graft outcomes., Methods: We studied by qPCR the presence of the DYS14 gene sequence of the Y chromosome in 12 females who received a kidney graft from a male donor before transplantation (T0), after 15 days (T1) and 1 year of transplantation (T2). We found the sequence in all recipients after renal transplantation., Results: All the women were negative for this sequence prior to transplantation (T0). Mean (SD) Y-related DNA quantity was 0.80 (0.69) ng/mL plasma and 0.15 (0.26) ng/mL plasma at T1 and T2, respectively. No acute rejection was observed, and mean (SD) estimated Cr clearance was 68.8 (16.9) mL/min within 1 year from transplantation., Conclusions: Presence of MC was associated with good kidney graft outcomes after 1 year of transplantation, but further studies will be needed to investigate the relationship between clinical outcomes and the development of MC in renal transplant recipient., (© 2020 S. Karger AG, Basel.)
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- 2020
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28. Analytical Evaluation of an NGS Testing Method for Routine Molecular Diagnostics on Melanoma Formalin-Fixed, Paraffin-Embedded Tumor-Derived DNA.
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Mancini I, Simi L, Salvianti F, Castiglione F, Sonnati G, and Pinzani P
- Abstract
Next Generation Sequencing (NGS) is a promising tool for the improvement of tumor molecular profiling in view of the identification of a personalized treatment in oncologic patients. To verify the potentiality of a targeted NGS (Ion AmpliSeq™ Cancer Hotspot Panel v2), selected melanoma samples ( n = 21) were retrospectively analyzed on S5 platform in order to compare NGS performance with the conventional techniques adopted in our routine clinical setting (Sequenom MassARRAY system, Sanger sequencing, allele-specific real-time PCR). The capability in the identification of rare and low-frequency mutations in the main genes involved in melanoma ( BRAF and NRAS genes) was verified and integrated with the results deriving from other oncogenes and tumor suppressor genes. The analytical evaluation was carried out by the analysis of DNA derived from control cell lines and FFPE (Formalin-Fixed, Paraffin-Embedded) samples to verify that the achieved resolution of uncommon mutations and low-frequency variants was suitable to meet the technical and clinical requests. Our results demonstrate that the amplicon-based NGS approach can reach the sensitivity proper of the allele-specific assays together with the high specificity of a sequencing method. An overall concordance among the tested methods was observed in the identification of classical and uncommon mutations. The assessment of the quality parameters and the comparison with the orthogonal methods suggest that the NGS method could be implemented in the clinical setting for melanoma molecular characterization.
- Published
- 2019
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29. Evaluation of the liquid biopsy for the detection of BRAFV600E mutation in metastatic melanoma patients.
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Salvianti F, Massi D, De Giorgi V, Gori A, Pazzagli M, and Pinzani P
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- Amino Acid Substitution, Biomarkers, Tumor genetics, Circulating Tumor DNA isolation & purification, DNA Mutational Analysis methods, Feasibility Studies, Glutamic Acid genetics, Humans, Liquid Biopsy methods, Melanoma blood, Melanoma pathology, Mutation, Real-Time Polymerase Chain Reaction, Skin Neoplasms blood, Skin Neoplasms pathology, Valine genetics, Biomarkers, Tumor blood, Circulating Tumor DNA blood, Melanoma diagnosis, Neoplastic Cells, Circulating pathology, Proto-Oncogene Proteins B-raf genetics, Skin Neoplasms diagnosis
- Abstract
Background: Circulating tumor cells (CTCs) and circulating cell free DNA (ccfDNA) represent a liquid biopsy of a tumor allowing real time disease monitoring especially in advanced stages of cancer, but their analysis is technically challenging., Objective: We aimed to demonstrate the feasibility of two different technical approaches to detect the BRAFV600E mutation in the liquid biopsy of 20 metastatic melanoma patients by using both the enriched CTC fraction and circulating ccfDNA from the same blood sample., Methods: We detected CTCs by a filtration method in 20 metastatic melanoma patients and detected the BRAFV600E variant on CTCs and ccfDNA by an allele-specific qPCR assay; the mutated samples were confirmed by ICE-COLD PCR followed by Sanger sequencing., Results: We found CTCs in 70% of the samples, and identified the BRAFV600E variant on CTCs. We correlated the results with those obtained on ccfDNA from the same blood draw. We found some discordant results between CTCs and ccfDNA., Conclusions: Our results underline the importance of investigating both CTCs and ccfDNA in a liquid biopsy approach to melanoma patients.
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- 2019
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30. The diagnostic potential of mutation detection from single circulating tumor cells in cancer patients.
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Salvianti F and Pinzani P
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- DNA Mutational Analysis standards, Genomics methods, Genomics standards, Humans, Liquid Biopsy, Sensitivity and Specificity, Single-Cell Analysis standards, Whole Genome Sequencing, Biomarkers, Tumor, DNA Mutational Analysis methods, Mutation, Neoplasms diagnosis, Neoplasms genetics, Neoplastic Cells, Circulating pathology, Single-Cell Analysis methods
- Abstract
Introduction: Circulating tumor cells (CTCs) have gained importance in the oncology field as biomarkers of tumor development. The most relevant observation that emerged from the recent studies on CTCs is their heterogeneity, which can be investigated by new technologies for single cell analysis. Areas covered: This review considers the most recent advances (limited to the last two years) in the mutational analysis of single CTCs with a critical point of view on the technical challenges still to be faced and the steps needed to reach a standardization of the procedures able to translate these new approaches into clinical practice. Expert commentary: CTCs represent a surrogate tumor sample obtained by a minimally invasive procedure allowing the serial monitoring of the patient during the follow-up period or after treatment. Notwithstanding that, the analysis of CTCs is not so widespread; in fact, a limited number of centers can be equipped and possess the expertise for the development of workflows able to identify, enrich and isolate CTCs from blood. Moreover, the lack of standardized procedures and guidelines limits the study of CTCs to 'research use only' approaches.
- Published
- 2017
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31. Circulating tumor cells and microemboli can differentiate malignant and benign pulmonary lesions.
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Mascalchi M, Maddau C, Sali L, Bertelli E, Salvianti F, Zuccherelli S, Matucci M, Borgheresi A, Raspanti C, Lanzetta M, Falchini M, Mazza E, Vella A, Luconi M, Pinzani P, and Pazzagli M
- Abstract
The presence of circulating tumor cells (CTC) or microemboli (CTM) in the peripheral blood can theoretically anticipate malignancy of solid lesions in a variety of organs. We aimed to preliminarily assess this capability in patients with pulmonary lesions of suspected malignant nature. We used a cell-size filtration method (ScreenCell) and cytomorphometric criteria to detect CTC/CTM in a 3 mL sample of peripheral blood that was taken just before diagnostic percutaneous CT-guided fine needle aspiration (FNA) or core biopsy of the suspicious lung lesion. At least one CTC/CTM was found in 47 of 67 (70%) patients with final diagnoses of lung malignancy and in none of 8 patients with benign pulmonary nodules. In particular they were detected in 38 (69%) of 55 primary lung cancers and in 9 (75%) of 12 lung metastases from extra-pulmonary cancers. Sensitivity of CTC/CTM presence for malignancy was 70.1% (95%CI: 56.9-83.1%), specificity 100%, positive predictive value 100% and negative predictive value 28.6% (95%CI: 11.9-45.3%). Remarkably, the presence of CTC/CTM anticipated the diagnosis of primary lung cancer in 3 of 5 patients with non-diagnostic or inconclusive results of FNA or core biopsy, whereas CTC/CTM were not observed in 1 patient with sarcoidosis and 1 with amarthocondroma. These results suggest that presently, due to the low sensitivity, the search of CTC/CTM cannot replace CT guided percutaneous FNA or core biopsy in the diagnostic work-up of patients with suspicious malignant lung lesions. However, the high specificity may as yet indicate a role in cases with non-diagnostic or inconclusive FNA or core biopsy results that warrants to be further investigated., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.
- Published
- 2017
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32. New insights in the clinical and translational relevance of miR483-5p in adrenocortical cancer.
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Salvianti F, Canu L, Poli G, Armignacco R, Scatena C, Cantini G, Di Franco A, Gelmini S, Ercolino T, Pazzagli M, Nesi G, Mannelli M, Pinzani P, and Luconi M
- Abstract
Adrenocortical cancer (ACC) is a rare aggressive malignancy. Recent ACC integrated genomics analysis contributed to redefine the risk groups on molecular basis, including tumor microRNAs (miRs), detectable also in the bloodstream. We developed a quantitative real-time (RT) assay for the measurement of miR483 and miR483-5p absolute levels in plasma samples. miR483/miR483-5p levels were evaluated in plasma samples of 27 patients with ACC before surgery and at follow-up. Statistically significant differences in miR483-5p and miR483 levels were found between stage 1/2 and stage 3/4 ACCs in pre-surgery and post-surgery samples. ROC curve analysis of miR483-5p levels gave a prediction of the clinical stage (accuracy 0.917±0.084), with the best cut-off value of 0.221 ng/ml, prognosticating overall and recurrence-free survival. In a multivariate Cox analysis (HR 16.2, 95%CI[1.39-188.6, P<0.026]), miR483-5p was the only variable that significantly predicted recurrence, but not overall survival. In addition, miR483 and miR483-5p levels correlated with the number of circulating tumor cells (CTCs) detected in the same blood samples, independently of the timing of sampling. In conclusion, we demonstrated that miR483-5p absolute plasma levels in ACC patients are powerful molecular markers that may help in the follow-up of patients after surgery and chemotherapy, and contribute to more accurately classify and predict tumor progression., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest
- Published
- 2017
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33. Integrity and Quantity of Total Cell-Free DNA in the Diagnosis of Thyroid Cancer: Correlation with Cytological Classification.
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Salvianti F, Giuliani C, Petrone L, Mancini I, Vezzosi V, Pupilli C, and Pinzani P
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- Adolescent, Adult, Aged, Aged, 80 and over, Case-Control Studies, DNA Fragmentation, Female, Humans, Liquid Biopsy, Male, Middle Aged, ROC Curve, Reproducibility of Results, Sensitivity and Specificity, Thyroid Neoplasms pathology, Thyroid Neoplasms surgery, Young Adult, Biomarkers, Tumor, Cell-Free Nucleic Acids, DNA, Neoplasm, Thyroid Neoplasms diagnosis, Thyroid Neoplasms genetics
- Abstract
Cell-free DNA (cfDNA) quantity and quality in plasma has been investigated as a non-invasive biomarker in cancer. Previous studies have demonstrated increased cfDNA amount and length in different types of cancer with respect to healthy controls. The present study aims to test the hypothesis that the presence of longer DNA strands circulating in plasma can be considered a biomarker for tumor presence in thyroid cancer. We adopted a quantitative real-time PCR (qPCR) approach based on the quantification of two amplicons of different length (67 and 180 bp respectively) to evaluate the integrity index 180/67. Cell-free DNA quantity and integrity were higher in patients affected by nodular thyroid diseases than in healthy controls. Importantly, cfDNA integrity index was higher in patients with cytological diagnosis of thyroid carcinoma (Thy4/Thy5) than in subjects with benign nodules (Thy2). Therefore, cfDNA integrity index 180/67 is a suitable parameter for monitoring cfDNA fragmentation in thyroid cancer patients and a promising circulating biomarker in the diagnosis of thyroid nodules., Competing Interests: The authors declare no conflict of interest.
- Published
- 2017
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34. Mutational analysis of single circulating tumor cells by next generation sequencing in metastatic breast cancer.
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De Luca F, Rotunno G, Salvianti F, Galardi F, Pestrin M, Gabellini S, Simi L, Mancini I, Vannucchi AM, Pazzagli M, Di Leo A, and Pinzani P
- Subjects
- Breast Neoplasms secondary, Female, Humans, Prognosis, Single-Cell Analysis, Survival Rate, Tumor Suppressor Protein p53 genetics, Biomarkers, Tumor genetics, Breast Neoplasms genetics, DNA Mutational Analysis methods, High-Throughput Nucleotide Sequencing methods, Mutation, Neoplastic Cells, Circulating pathology
- Abstract
Circulating Tumor Cells (CTCs) represent a "liquid biopsy" of the tumor potentially allowing real-time monitoring of cancer biology and therapies in individual patients.The purpose of the study was to explore the applicability of a protocol for the molecular characterization of single CTCs by Next Generation Sequencing (NGS) in order to investigate cell heterogeneity and provide a tool for a personalized medicine approach.CTCs were enriched and enumerated by CellSearch in blood from four metastatic breast cancer patients and singularly isolated by DEPArray. Upon whole genome amplification 3-5 single CTCs per patient were analyzed by NGS for 50 cancer-related genes.We found 51 sequence variants in 25 genes. We observed inter- and intra-patient heterogeneity in the mutational status of CTCs.The highest number of somatic deleterious mutations was found in the gene TP53, whose mutation is associated with adverse prognosis in breast cancer.The discordance between the mutational status of the primary tumor and CTCs observed in 3 patients suggests that, in advanced stages of cancer, CTC characteristics are more closely linked to the dynamic modifications of the disease status.In one patient the mutational profiles of CTCs before and during treatment shared only few sequence variants.This study supports the applicability of a non-invasive approach based on the liquid biopsy in metastatic breast cancer patients which, in perspective, should allow investigating the clonal evolution of the tumor for the development of new therapeutic strategies in precision medicine., Competing Interests: The authors declare no potential conflicts of interest.
- Published
- 2016
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35. Tumor-Related Methylated Cell-Free DNA and Circulating Tumor Cells in Melanoma.
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Salvianti F, Orlando C, Massi D, De Giorgi V, Grazzini M, Pazzagli M, and Pinzani P
- Abstract
Solid tumor release into the circulation cell-free DNA (cfDNA) and circulating tumor cells (CTCs) which represent promising biomarkers for cancer diagnosis. Circulating tumor DNA may be studied in plasma from cancer patients by detecting tumor specific alterations, such as genetic or epigenetic modifications. Ras association domain family 1 isoform A (RASSF1A) is a tumor suppressor gene silenced by promoter hypermethylation in a variety of human cancers including melanoma. The aim of the present study was to assess the diagnostic performance of a tumor-related methylated cfDNA marker in melanoma patients and to compare this parameter with the presence of CTCs. RASSF1A promoter methylation was quantified in cfDNA by qPCR in a consecutive series of 84 melanoma patients and 68 healthy controls. In a subset of 68 cases, the presence of CTCs was assessed by a filtration method (Isolation by Size of Epithelial Tumor Cells, ISET) as well as by an indirect method based on the detection of tyrosinase mRNA by RT-qPCR. The distribution of RASSF1A methylated cfDNA was investigated in cases and controls and the predictive capability of this parameter was assessed by means of the area under the ROC curve (AUC). The percentage of cases with methylated RASSF1A promoter in cfDNA was significantly higher in each class of melanoma patients (in situ, invasive and metastatic) than in healthy subjects (Pearson chi-squared test, p < 0.001). The concentration of RASSF1A methylated cfDNA in the subjects with a detectable quantity of methylated alleles was significantly higher in melanoma patients than in controls. The biomarker showed a good predictive capability (in terms of AUC) in discriminating between melanoma patients and healthy controls. This epigenetic marker associated to cfDNA did not show a significant correlation with the presence of CTCs, but, when the two parameters are jointly considered, we obtain a higher sensitivity of the detection of positive cases in invasive and metastatic melanomas. Our data suggest that cell-free tumor DNA and CTCs represent two complementary aspects of the liquid biopsy which may improve the diagnosis and the clinical management of melanoma patients.
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- 2016
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36. Prevalence and number of circulating tumour cells and microemboli at diagnosis of advanced NSCLC.
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Mascalchi M, Falchini M, Maddau C, Salvianti F, Nistri M, Bertelli E, Sali L, Zuccherelli S, Vella A, Matucci M, Voltolini L, Pegna AL, Luconi M, Pinzani P, and Pazzagli M
- Subjects
- Adenocarcinoma metabolism, Aged, Aged, 80 and over, Biopsy, Fine-Needle, Carcinoma, Non-Small-Cell Lung metabolism, Carcinoma, Squamous Cell metabolism, Female, Follow-Up Studies, Humans, Lung Neoplasms metabolism, Lymphatic Metastasis, Male, Middle Aged, Neoplasm Staging, Prognosis, Vimentin metabolism, Adenocarcinoma secondary, Biomarkers, Tumor blood, Carcinoma, Non-Small-Cell Lung secondary, Carcinoma, Squamous Cell secondary, Lung Neoplasms pathology, Neoplastic Cells, Circulating pathology
- Abstract
Purpose: Timing and magnitude of blood release of circulating tumour cells (CTC) and circulating tumour microemboli (CTM) from primary solid cancers are uncertain. We investigated prevalence and number of CTC and CTM at diagnosis of advanced non-small cell lung cancer (NSCLC)., Methods: Twenty-eight consecutive patients with suspected stage III-IV lung cancer gave consent to provide 15 mL of peripheral blood soon before diagnostic CT-guided fine-needle aspiration biopsy (FNAB). CTC and CTM (clusters of ≥3 CTC) were isolated by cell size filtration (ScreenCell), identified and counted by cytopathologists using morphometric criteria and (in 6 cases) immunostained for vimentin., Results: FNAB demonstrated NSCLC in 26 cases. At least one CTC/3 mL blood (mean 6.8 ± 3.7) was detected in 17 (65 %) and one CTM (mean 4.5 ± 3.3) in 15 (58 %) of 26 NSCLC cases. No correlation between number of CTC or CTM and tumour type or stage was observed. Neoplastic cells from both FNA and CTC/CTM were positive for vimentin but heterogeneously., Conclusions: CTC can be detected in two-thirds and CTM in more than half of patients with advanced NSCLC at diagnosis. Reasons underlying lack of CTC and CTM in some advanced lung cancers deserve further investigations.
- Published
- 2016
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37. Single circulating tumor cell sequencing as an advanced tool in cancer management.
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Salvianti F, Pazzagli M, and Pinzani P
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- Biopsy, Humans, Mutation, Neoplasm Metastasis, Neoplasms genetics, Neoplasms pathology, Neoplastic Cells, Circulating metabolism, Neoplasms blood, Neoplastic Cells, Circulating pathology, Sequence Analysis, DNA, Single-Cell Analysis
- Abstract
Circulating tumor cells (CTCs) shed by the primary tumor and metastases are considered a real-time 'liquid biopsy', reflecting the disease complexity that evolves during progression, showing in its late stages different genetic, epigenetic and expression features. Consequently, heterogeneity and development of characteristic features upon disease progression are the two main goals that emerging technologies should account for in view of a clinical application. Single-cell analysis, now possible due to technological advances, may help elucidate tumor heterogeneity at the CTC level. This review focuses on the necessary steps for the analysis of CTCs at the single-cell level. A concise overview is given on the alternative methods referring in particular to studies on the mutational status of single CTCs from cancer patients.
- Published
- 2016
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38. Feasibility of a workflow for the molecular characterization of single cells by next generation sequencing.
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Salvianti F, Rotunno G, Galardi F, De Luca F, Pestrin M, Vannucchi AM, Di Leo A, Pazzagli M, and Pinzani P
- Abstract
The purpose of the study was to explore the feasibility of a protocol for the isolation and molecular characterization of single circulating tumor cells (CTCs) from cancer patients using a single-cell next generation sequencing (NGS) approach. To reach this goal we used as a model an artificial sample obtained by spiking a breast cancer cell line (MDA-MB-231) into the blood of a healthy donor. Tumor cells were enriched and enumerated by CellSearch(®) and subsequently isolated by DEPArray™ to obtain single or pooled pure samples to be submitted to the analysis of the mutational status of multiple genes involved in cancer. Upon whole genome amplification, samples were analysed by NGS on the Ion Torrent PGM™ system (Life Technologies) using the Ion AmpliSeq™ Cancer Hotspot Panel v2 (Life Technologies), designed to investigate genomic "hot spot" regions of 50 oncogenes and tumor suppressor genes. We successfully sequenced five single cells, a pool of 5 cells and DNA from a cellular pellet of the same cell line with a mean depth of the sequencing reaction ranging from 1581 to 3479 reads. We found 27 sequence variants in 18 genes, 15 of which already reported in the COSMIC or dbSNP databases. We confirmed the presence of two somatic mutations, in the BRAF and TP53 gene, which had been already reported for this cells line, but also found new mutations and single nucleotide polymorphisms. Three variants were common to all the analysed samples, while 18 were present only in a single cell suggesting a high heterogeneity within the same cell line. This paper presents an optimized workflow for the molecular characterization of multiple genes in single cells by NGS. The described pipeline can be easily transferred to the study of single CTCs from oncologic patients.
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- 2015
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39. Prospective evaluation of RASSF1A cell-free DNA as a biomarker of pre-eclampsia.
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Salvianti F, Inversetti A, Smid M, Valsecchi L, Candiani M, Pazzagli M, Cremonesi L, Ferrari M, Pinzani P, and Galbiati S
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- Adult, Biomarkers blood, DNA blood, DNA chemistry, DNA Methylation, Epidemiologic Studies, Female, Humans, Predictive Value of Tests, Pregnancy, Promoter Regions, Genetic, Pre-Eclampsia blood, Tumor Suppressor Proteins genetics
- Abstract
Introduction: This study aims to quantify total and fetal cell-free DNA (cfDNA) in maternal plasma at different gestational ages and to assess whether this could represent a reliable predictive marker of pre-eclampsia (PE) before clinical onset., Methods: We performed a qPCR assay to compare the cfDNA concentration of hypermethylated and unmethylated RASSF1A promoter gene sequences in maternal plasma among 3 groups of pregnant women. These included 17 women with overt PE, 33 women at risk for the disease subsequently differentiated into 9 who developed PE and 24 who did not, and 73 controls. All women at risk were consecutively sampled throughout the whole gestation., Results: Both total and fetal cfDNA had a good diagnostic performance in distinguishing patients with overt PE from healthy controls. When comparing women at risk who developed PE to women at risk who did not, the predictive capability was satisfactory at a gestational age ranging from 17 to 30 weeks. This allowed establishing within this time interval a cut-off value of 735 GE/ml for total cfDNA (87.5% sensitivity and 70.0% specificity), and a cut-off value of 7.49 GE/ml for fetal cfDNA (100% sensitivity and 50% specificity). cfDNA levels turned positive several weeks before the onset of the disease: from 2 to 18 weeks for total cfDNA and from 8 to 17 weeks for fetal cfDNA., Discussion: The simultaneous use of total and fetal cfDNA would allow an accurate monitoring and prevention of PE development thus suggesting that RASSF1A could represent a potential biomarker of PE., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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40. Heterogeneity of PIK3CA mutational status at the single cell level in circulating tumor cells from metastatic breast cancer patients.
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Pestrin M, Salvianti F, Galardi F, De Luca F, Turner N, Malorni L, Pazzagli M, Di Leo A, and Pinzani P
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- Adult, Aged, Base Sequence, Breast Neoplasms pathology, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases, DNA Mutational Analysis, Female, Humans, Middle Aged, Molecular Sequence Data, Neoplasm Metastasis, Reproducibility of Results, Breast Neoplasms enzymology, Breast Neoplasms genetics, Genetic Heterogeneity, Neoplastic Cells, Circulating pathology, Phosphatidylinositol 3-Kinases genetics, Single-Cell Analysis
- Abstract
Circulating Tumor Cells (CTCs) represent a "liquid biopsy of the tumor" which might allow real-time monitoring of cancer biology and therapies in individual patients. CTCs are extremely rare in the blood stream and their analysis is technically challenging. The CellSearch(®) system provides the enumeration of CTCs with prognostic significance in patients with metastatic breast cancer (mBC), but it does not allow their molecular characterization, which might be useful to identify therapeutically relevant targets for individualized treatment. Combining the CellSearch(®) and DEPArray™ technologies allows the recovery of single CTCs as a pure sample for molecular analysis. The purpose of the study was to investigate the heterogeneity of PIK3CA mutational status within single CTCs isolated from individual mBC patients. CTCs were enriched and enumerated by CellSearch(®) in blood samples collected from 39 mBC patients. In 20 out of 39 patients enriched samples with ≥5 CTCs were sorted using DEParray™ to isolate single CTCs or pools of CTCs to be submitted to Whole Genome Amplification (WGA) before sequencing analysis. In 18 out of 20 patients, it was possible to perform PIK3CA sequencing on exons 9 and 20. Twelve subjects were wild type (wt) for the PIK3CA gene. PIK3CA status could also be assessed in pools of CTCs in seven of these patients, with consistent wt status found. Six patients (33%) had a PIK3CA mutation identified. In 2 of the six patients, molecular heterogeneity was detected when mutational analysis was performed on more than one single CTC, including one patient with loss of heterozygosity on both single and pooled CTCs, and one patient with three different PIK3CA variants on single CTCs but PIK3CA wt status on pooled CTC samples. In six out of the 18 cases PIK3CA status was also evaluable on a primary tumor sample. In one of the six cases a discordance in PIK3CA status between the primary (wild-type) and the matched CTC (exon 20 mutation) was observed. This study demonstrates the feasibility of a non-invasive approach based on the liquid biopsy in mBC patients. Moreover, our data suggest the importance of characterizing CTCs at the single cell level in order to investigate the molecular heterogeneity within cells from the same patient., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2015
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41. Atypical Spitz tumors in patients younger than 18 years.
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Massi D, Tomasini C, Senetta R, Paglierani M, Salvianti F, Errico ME, Donofrio V, Collini P, Tragni G, Sementa AR, Rongioletti F, Boldrini R, Ferrari A, Gambini C, and Montesco MC
- Subjects
- Adolescent, Child, Child, Preschool, Female, Humans, Infant, Male, Retrospective Studies, Nevus, Epithelioid and Spindle Cell pathology, Skin Neoplasms pathology
- Abstract
Background: Diagnosis and proper management of atypical Spitz tumors in pediatric age are still controversial., Objective: We sought to investigate the clinicopathological and molecular features of atypical Spitz tumors in patients aged 18 years or younger., Methods: We performed a retrospective clinicopathological and fluorescence in situ hybridization study on 50 pediatric atypical Spitz tumors., Results: Parameters that were significantly correlated with a diagnosis of atypical Spitz tumors over Spitz nevus included asymmetry, level IV/V, lack of maturation, solid growth, nuclear pleomorphism, high nuclear-cytoplasmic ratio, atypical and deep mitoses, and more than 6 mitoses/mm(2). In the atypical Spitz tumors group, a significantly higher mitotic rate was observed in prepuberal age (P = .04). The 4-probe fluorescence in situ hybridization melanoma assay did not discriminate atypical Spitz tumors from Spitz nevi. Heterozygous 9p21 loss was found in 3 of 37 cases and homozygous 9p21 loss in 2 of 37 cases. Only 1 child experienced a fatal outcome, showing genetic abnormalities by melanoma fluorescence in situ hybridization probe and a heterozygous 9p21 deletion., Limitations: The limited number of adverse outcomes did not allow the prognostic analysis of single morphologic features., Conclusion: Pediatric atypical Spitz tumors are associated with minimal lethal potential. Atypical Spitz tumors require complete excision and careful follow-up while our data do not support any clinical benefit for the sentinel lymph node biopsy procedure and completion lymphadenectomy., (Copyright © 2014 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.)
- Published
- 2015
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42. Synchronous occurrence of medullary and papillary carcinoma of the thyroid in a patient with cutaneous melanoma: determination of BRAFV600E in peripheral blood and tissues. Report of a case and review of the literature.
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Fibbi B, Pinzani P, Salvianti F, Rossi M, Petrone L, De Feo ML, Panconesi R, Vezzosi V, Bianchi S, Simontacchi G, Mangoni M, Pertici M, Forti G, and Pupilli C
- Subjects
- Alleles, Carcinoma, Medullary genetics, Carcinoma, Papillary genetics, DNA Mutational Analysis, Humans, Male, Middle Aged, Mutation, Neoplasms, Multiple Primary genetics, Proto-Oncogene Mas, Thyroid Neoplasms genetics, Carcinoma, Medullary pathology, Carcinoma, Papillary pathology, Neoplasms, Multiple Primary pathology, Proto-Oncogene Proteins B-raf genetics, Thyroid Gland pathology, Thyroid Neoplasms pathology
- Abstract
The purpose of this study is to describe a case of concurrent medullary and papillary thyroid carcinoma (MTC and PTC) and cutaneous melanoma and to analyze BRAF(V600E) mutation in plasma and tissues. We report the clinical history and the laboratory, imaging, and histopathological findings of a 47-year-old man affected by multinodular goiter. BRAF(V600E)-mutated DNA was quantified in plasma samples and in cancer sections by quantitative real-time polymerase chain reaction (qPCR). At ultrasound examination, the dominant right nodule of the thyroid was weakly hyperechoic and hypervascularized, while the left one was hypoechoic without internal vascularization. Regional lymphadenomegalia was not detected. Basal plasma calcitonin was elevated, and the patient underwent total thyroidectomy and resection of central cervical lymph nodes. Histopathological examination identified two distinct foci of MTC and PTC and micrometastasis of well-differentiated carcinoma in one of the six resected lymph nodes. RET proto-oncogene germline mutations were not detected. Cutaneous melanoma of the thorax was subsequently diagnosed. BRAF(V600E) tissue DNA was detected in PTC and melanoma but not in MTC. The cell-free plasma percentage of BRAF(V600E) DNA was detected in pre-thyroidectomy peripheral blood and was drastically reduced after cancer treatments. This study confirms the occurrence of synchronous MTC and PTC and is the first evidence of the co-existence of melanoma and distinct thyroid cancers of different origin. BRAF(V600E) allele was detected in PTC and melanoma but not in MTC tissues. BRAF(V600E) molecular quantification in pre- and post-treatment blood supports our previous data, suggesting its possible role in diagnosis and follow-up of BRAF-positive tumors.
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- 2014
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43. Circulating tumor cells detection and counting in uveal melanomas by a filtration-based method.
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Mazzini C, Pinzani P, Salvianti F, Scatena C, Paglierani M, Ucci F, Pazzagli M, and Massi D
- Abstract
Uveal melanoma is one of the most deadly diseases in ophthalmology for which markers able to predict the appearance of metastasis are needed. The study investigates the role of circulating tumor cells (CTC) as a prognostic factor in this disease. We report the detection of circulating tumor cells by Isolation by Size of Epithelial Tumor cells (ISET) in a cohort of 31 uveal melanoma patients: we identified single CTCs or clusters of cells in 17 patients, while the control population, subjects with choroidal nevi, showed no CTC in peripheral blood. The presence of CTCs did not correlate with any clinical and pathological parameter, such as tumor larger basal diameter (LBD), tumor height and TNM. By stratifying patients in groups on the basis of the number of CTC (lower or higher than 10 CTC per 10 mL blood) and the presence of CTC clusters we found a significant difference in LBD (p = 0.019), Tumor height (p = 0.048), disease-free and overall survival (p < 0.05). In conclusion, we confirm the role of CTC as a negative prognostic marker in uveal melanoma patients after a long follow-up period. Further characterization of CTC will help understanding uveal melanoma metastasization and improve patient management.
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- 2014
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44. Circulating cell-free DNA in cancer.
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Pinzani P, Salvianti F, Orlando C, and Pazzagli M
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- Alleles, Blood Specimen Collection, Cell-Free System metabolism, DNA, Neoplasm genetics, Humans, Mutation, DNA, Neoplasm blood, Neoplasms blood, Polymerase Chain Reaction methods
- Abstract
This papers deals with the preanalytical and analytical phase of cell-free DNA analysis, highlighting some criticism on sample collection and extraction. We describe a method to accurately quantify total cfDNA in plasma and our particular approach to the measurement of tumor deriving cfDNA.
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- 2014
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45. BRAF and KIT somatic mutations are present in amelanotic melanoma.
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Massi D, Pinzani P, Simi L, Salvianti F, De Giorgi V, Pizzichetta MA, Mirri F, Steffan A, Orlando C, Santucci M, and Canzonieri V
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- Adult, Aged, Aged, 80 and over, DNA Mutational Analysis methods, Female, Gene Frequency, Genetic Predisposition to Disease, Humans, Italy, Male, Melanoma, Amelanotic enzymology, Melanoma, Amelanotic pathology, Middle Aged, Phenotype, Real-Time Polymerase Chain Reaction, Retrospective Studies, Risk Factors, Skin Neoplasms enzymology, Skin Neoplasms pathology, Biomarkers, Tumor genetics, Melanoma, Amelanotic genetics, Mutation, Proto-Oncogene Proteins B-raf genetics, Proto-Oncogene Proteins c-kit genetics, Skin Neoplasms genetics
- Abstract
The genotypic profile of rare amelanotic melanomas (AMs) has been poorly investigated, thus preventing either an accurate identification as a distinctive melanoma subtype or therapy stratification. Here, we investigated the presence of the BRAF(V600E) mutation by real-time quantitative PCR and KIT mutations (exons 11 and 17) by sequencing analysis in 33 AMs. AMs included 'truly' amelanotic lesions (n = 19), with no melanin pigmentation upon dermoscopic inspection and hypomelanotic lesions (n = 14), by definition partially pigmented lesions showing a melanin pigmentation area of less than 25% of the total surface area. The frequency of the BRAF(V600E) mutation was 70.3% in the 33 cases, a percentage that increased to 89% when only the subgroup of thin melanomas (≤ 1 mm in thickness, n = 9) was considered. KIT mutations were found in 12.1% of AMs, all of which developed in nonacral sites. The identification of a relatively high frequency of BRAF(V600E) and KIT mutations in AMs may have important consequences for implementation of the novel targeted therapies now available to treat this life-threatening disease.
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- 2013
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46. Detection of circulating tumor cells in patients with adrenocortical carcinoma: a monocentric preliminary study.
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Pinzani P, Scatena C, Salvianti F, Corsini E, Canu L, Poli G, Paglierani M, Piccini V, Pazzagli M, Nesi G, Mannelli M, and Luconi M
- Subjects
- Adult, Disease Progression, Female, Humans, Male, Middle Aged, Prognosis, Adrenal Cortex Neoplasms pathology, Adrenocortical Carcinoma pathology, Neoplastic Cells, Circulating pathology
- Abstract
Context: Adrenocortical carcinoma (ACC) is a rare malignancy, the prognosis of which is mainly dependent on stage at diagnosis. The identification of disease-associated markers for early diagnosis and drug monitoring is mandatory. Circulating tumor cells (CTCs) are released into the bloodstream from primary tumor/metastasis. CTC detection in blood samples may have enormous potential for assisting in the diagnosis of malignancy, estimating prognosis, and monitoring the disease., Objective: The aim of the study was to investigate the presence of CTCs in blood samples of patients with ACC or benign adrenocortical adenoma (ACA)., Setting: We conducted the study at a university hospital., Intervention: CTC analysis was performed in blood samples from 14 ACC patients and 10 ACA patients. CTCs were isolated on the basis of cell size by filtration through ScreenCell devices, followed by identification according to validated morphometric criteria and immunocytochemistry., Main Outcome Measure: We measured the difference in CTC detection between ACC and ACA., Results: CTCs were detected in all ACC samples, but not in ACA samples. Immunocytochemistry confirmed the adrenocortical origin. When ACC patients were stratified according to the median value of tumor diameter and metastatic condition, a statistically significant difference was found in the number of CTCs detected after surgery. A significant correlation between the number of CTCs in postsurgical samples and clinical parameters was found for tumor diameter alone., Conclusions: Our findings provide the first evidence for adrenocortical tumors that CTCs may represent a useful marker to support differential diagnosis between ACC and ACA. The correlation with some clinical parameters suggests a possible relevance of CTC analysis for prognosis and noninvasive monitoring of disease progression and drug response.
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- 2013
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47. Cell-free fetal DNA in maternal circulation after chorionic villous sampling.
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Di Tommaso M, Seravalli V, Salvianti F, Bussani C, Pasquini L, Cordisco A, and Pinzani P
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- DNA Methylation, Female, Fetal Blood chemistry, Fetomaternal Transfusion blood, Fetomaternal Transfusion etiology, Gestational Age, Humans, Maternal Age, Parity, Placenta chemistry, Pregnancy, Promoter Regions, Genetic genetics, Real-Time Polymerase Chain Reaction, Tumor Suppressor Proteins genetics, Chorionic Villi Sampling adverse effects, DNA blood
- Abstract
Objective: This study aims to estimate whether chorionic villous sampling (CVS) causes a significant increase of cell-free fetal DNA (cffDNA) in maternal circulation., Method: Fifty pregnant women with singleton pregnancy were recruited prior to CVS. Maternal peripheral blood was collected before and after CVS. A methylation-sensitive restriction enzyme digestion was used to select the placental-derived hypermethylated promoter of the RASSF1A gene in maternal plasma, thus differentiating cffDNA from mother's cell-free DNA (cfDNA), where the RASSF1A gene is normally hypomethylated. Total cfDNA and cffDNA amounts were compared before and after CVS in each patient. Data were compared using the Student t-test., Results: No significant difference before and after CVS was found between the following: (i) total cfDNA concentration in plasma (p = 0.695); (ii) cffDNA concentration in plasma (p = 0.612); and (iii) percentage of fetal DNA in plasma (p = 0.835). After dividing the cases on the basis of the sex of the fetus, maternal age, gestational age, number of pregnancies, position of the placenta, and presence of trisomy of the fetus, no difference in fetal and total DNA concentrations before and after CVS was observed., Conclusion: The CVS does not seem to significantly disrupt the maternal-placental interface, as no significant increase of cffDNA in maternal plasma following CVS was observed., (© 2013 John Wiley & Sons, Ltd.)
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- 2013
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48. Sustained proliferation and resistance to apoptosis after a cytotoxic insult are early alterations in rat colon carcinogenesis.
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Femia AP, Salvianti F, Luceri C, Dolara P, Salvadori M, Pinzani P, and Caderni G
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- 1,2-Dimethylhydrazine pharmacology, Animals, Aspirin pharmacology, Biomarkers, Tumor, Caspase 3 metabolism, Cell Proliferation, Colonic Neoplasms genetics, Dinoprostone blood, Interleukin-1beta blood, Intestinal Mucosa metabolism, Male, Microtubule-Associated Proteins genetics, Mucins metabolism, Precancerous Conditions genetics, Precancerous Conditions metabolism, Rats, Rats, Inbred F344, Survivin, bcl-2 Homologous Antagonist-Killer Protein metabolism, Apoptosis drug effects, Cell Transformation, Neoplastic pathology, Colonic Neoplasms metabolism, Colonic Neoplasms pathology, Intestinal Mucosa pathology, Microtubule-Associated Proteins metabolism, Precancerous Conditions pathology
- Abstract
To study the early alterations in carcinogenesis, we determined apoptosis and proliferation in rat mucin depleted foci (MDF), precancerous lesions in the colon under basal conditions and 24 h after treatment with 1,2-dimethylhydrazine (DMH), which induces apoptosis in the colon. Spontaneous apoptosis in MDF was higher than in normal mucosa (Apoptotic Index was 1.61 ± 0.30 and 0.21 ± 0.02 in MDF and normal mucosa, respectively, mean ± SE, p < 0.05). DMH (30 and 75 mg/kg) increased apoptosis in both normal mucosa and MDF (up to 20 times higher compared to basal levels in normal mucosa, but only two times in MDF). MDF had a higher and deregulated pattern of proliferation along the crypt compared to normal mucosa. After DMH, proliferation in normal mucosa was significantly depressed, but it did not vary in MDF. Survivin-Birc5 regulating apoptosis and proliferation was significantly over-expressed (RT-qPCR and immunohistochemistry experiments) in MDF vs. normal mucosa, but did not vary in response to DMH. The expression of the pro-apoptotic protein Bak did not vary in normal mucosa and MDF. Since inflammation is present in MDF, which may hamper apoptosis, we studied the effect of pre-treatment with aspirin (600 ppm in the diet for 10 days). No significant effects of aspirin were observed. In conclusion, MDF had a higher spontaneous apoptosis and proliferation coupled with a reduced response to apoptotic stimuli from cytotoxic compounds. Survivin over-expression in MDF indicates that this is an early event in colon carcinogenesis and suggests that down-regulation of Survivin may represent a strategy for cancer prevention., (Copyright © 2011 UICC.)
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- 2012
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49. Androgen receptor (AR) expression in prostate cancer and progression of the tumor: Lessons from cell lines, animal models and human specimens.
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Tamburrino L, Salvianti F, Marchiani S, Pinzani P, Nesi G, Serni S, Forti G, and Baldi E
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- Animals, Cell Line, Tumor, Disease Models, Animal, ErbB Receptors metabolism, Humans, Male, Neoplasms, Hormone-Dependent therapy, Orchiectomy, Prostatic Neoplasms pathology, Prostatic Neoplasms therapy, Receptors, Androgen genetics, Neoplasms, Hormone-Dependent metabolism, Neoplasms, Hormone-Dependent pathology, Prostatic Neoplasms metabolism, Receptors, Androgen metabolism
- Abstract
Prostate cancer (PC) is among the most frequent causes of death for cancer in men in western countries. In about 30% of cases, the disease is very aggressive rapidly leading to a metastatic disease. In these cases, prostatectomy is not possible and the patient is usually directed to androgen deprivation therapy (ADT) which is only palliative as a castration resistant PC (CRPC) usually develops within 2-3 years of treatment. At present there are no prognostic markers of PC progression. The role of the androgen receptor (AR) in initiation and development of PC is well established and documented. In particular, it is now recognized that androgens actions are mediated by an integration of classical (genomic) and non-classical (extragenomic) activity of AR. The picture about AR and PC become less clear when CRPC is considered. Indeed, the role of AR in the progression of PC and in CRPC is controversial. Results of studies on the role of AR in the progression of PC in cell lines, xenografts, animal models and even clinical specimens are conflicting reflecting the high heterogeneity of PC. Recent evidence in AR conditional KO in mouse models of PC shows possible contrasting roles of AR depending on its location in the two (epithelial or stromal) compartments of PC. Here, we review this evidence and report preliminary data of a study performed in microdissected areas of epithelia and stromal compartments of human PC., (Copyright © 2012 Elsevier Inc. All rights reserved.)
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- 2012
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50. Marie Ménard apples with high polyphenol content and a low-fat diet reduce 1,2-dimethylhydrazine-induced colon carcinogenesis in rats: effects on inflammation and apoptosis.
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Femia AP, Luceri C, Bianchini F, Salvadori M, Salvianti F, Pinzani P, Dolara P, Calorini L, and Caderni G
- Subjects
- 1,2-Dimethylhydrazine adverse effects, Animals, Anti-Infective Agents pharmacology, Apoptosis drug effects, Colitis pathology, Colon drug effects, Colon metabolism, Colonic Neoplasms chemically induced, Gene Expression Regulation drug effects, Interleukin-1beta genetics, Interleukin-6 genetics, Male, Precancerous Conditions chemically induced, Precancerous Conditions pathology, Precancerous Conditions prevention & control, Proliferating Cell Nuclear Antigen genetics, Rats, Rats, Inbred F344, Tumor Necrosis Factor-alpha genetics, Colitis drug therapy, Colonic Neoplasms pathology, Colonic Neoplasms prevention & control, Diet, Fat-Restricted, Malus chemistry, Polyphenols pharmacology
- Abstract
Inflammation may increase cancer risk, therefore, we studied whether polyphenol-rich Marie Ménard (MM) apples with reported anti-inflammatory activity prevent 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats and, likewise whether high-fat (HF) diet promoting carcinogenesis, may affect inflammation. DMH-induced rats were fed for 15 weeks with: an HF diet (23% corn oil w/w); an HF diet containing 7.6% w/w lyophilized MM (apple diet (AD)); a low-fat (LF) diet and an HF diet containing piroxicam (PXC) (0.01% w/w) as control. Mucin depleted foci (MDF), precancerous lesions in the colon, were dramatically reduced in the AD, LF, and PXC groups compared with the HF. Peritoneal macrophage activation, an index of systemic inflammation, was significantly decreased in the AD, LF, and PXC groups. TNF-α, iNOS, IL-1β, IL-6 m-RNA expression in the colon, as well as CD68 cells and plasmatic PGE2 were lower in the AD, but not in the LF group. Apoptosis in the MDF of both the AD and LF-fed rats was significantly higher than in HF rats. In conclusion, AD has a strong chemopreventive effect, reducing inflammation, and increasing apoptosis, while the chemopreventive effect of the LF diet seems mediated mainly by increased apoptosis in MDF., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2012
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