Luca REGGIANI BONETTI - personale UniMoRe

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Luca REGGIANI BONETTI

Professore Associato
Dipartimento di Scienze Mediche e Chirurgiche Materno-Infantili e dell'Adulto

Pubblicazioni

2024 - Dermoscopic, Histological, Confocal Microscopy Correlation of Atypical-Dysplastic Melanocytic Nevi [Articolo su rivista]
Cantisani, Carmen; Ambrosio, Luca; Annessi, Emanuele; Longo, Caterina; Farnetani, Francesca; Pezzini, Claudia; Condorelli, Alessandra; Annessi, Giorgio; Bonetti, Luca Reggiani; Guida, Stefania; Cota, Carlo; Tammaro, Antonella; Chello, Camilla; Pellacani, Giovanni
abstract

Introduction: The term "atypical melanocytic nevus" (AMN) is used as a synonym for dysplastic nevus (DN) in clinical practice. Although the criteria for diagnosis of AMN/DN by the Agency for Research on Cancer helps to differentiate AMN/DN from common acquired nevi, they do not have high degrees of specificity, as they are similar to those used for the diagnosis of melanoma. Objectives: In this retrospective study we evaluated the correlation and diagnostic concordance of dermoscopy, confocal microscopy, and histological examination in 50 AMN. Methods: A graded scale was used to compare histological examination with dermoscopy and confocal microscopy. Low magnification histological images of only the central part of lesions were examined. This allowed histological diagnoses based almost exclusively on architectural criteria instead of simultaneously architectural and cytological, as in the global histological examination. Results: Our data demonstrate that the diagnostic accuracy of dermoscopy and confocal microscopy diagnosis of the clinical aspects of AMN/DN as nevi or melanomas tends to be equivalent, being fair for nevi and excellent for melanomas. The total percentage of AMN suggested that the accuracy of confocal microscopy in the diagnosis of melanoma (86.7%) is greater than that of dermoscopy (73.3%). Conclusions: This study demonstrated that diagnostic assessments of AMN/DN by dermoscopy and confocal microscopy are accurate and often coincide with those of histological examination and that their combined use helps to better manage and monitor these patients by facilitating early detection of melanomas and reducing unnecessary excisions of benign melanocytic lesions.

2023 - Artificial intelligence evaluation of confocal microscope prostate images: our preliminary experience [Articolo su rivista]
Bianchi, G.; Puliatti, S.; Rodriguez, N.; Micali, S.; Bertoni, L.; Reggiani Bonetti, L.; Caramaschi, S.; Bolelli, F.; Pinamonti, M.; Rozze, D.; Grana, C.
abstract

2023 - CD271 activation prevents low to high-risk progression of cutaneous squamous cell carcinoma and improves therapy outcomes [Articolo su rivista]
Quadri, Marika; Tiso, Natascia; Musmeci, Francesco; I Morasso, Maria; R Brooks, Stephen; REGGIANI BONETTI, Luca; Panini, Rossana; Lotti, Roberta; Marconi, Alessandra; Pincelli, Carlo; Palazzo, Elisabetta
abstract

2023 - Clinical Significance of Molecular Subtypes in Western Advanced Gastric Cancer: A Real-World Multicenter Experience [Articolo su rivista]
Salati, Massimiliano; Ghidini, Michele; Paccagnella, Matteo; Reggiani Bonetti, Luca; Bocconi, Alessandro; Spallanzani, Andrea; Gelsomino, Fabio; Barbin, Francesca; Garrone, Ornella; Daniele, Bruno; Dominici, Massimo; Facciorusso, Antonio; Petrillo, Angelica
abstract

: In recent years, the molecular subtyping of gastric cancer has led to the identification of novel clinically relevant biomarkers as well as promising therapeutic targets. In parallel, the advent of checkpoint inhibitors has expanded treatment options beyond conventional chemotherapy. Compelling evidence has shown unprecedented efficacy results for anti-PD1-based therapies in the molecular subgroups of dMMR/MSI-h, EBV+ and PD-L1 CPS+ patients, to the point that these are granted approval for gastric cancer adenocarcinoma (AGC) in several countries. Despite this, cytotoxic chemotherapy remains the only treatment choice for the considerable proportion of biomarkers-negative patients. In this context, little is known about the association between subtypes-defining biomarkers (HER2, MMR/MSI, PD-L1, and EBV) and the efficacy of standard chemotherapy in non-Asian AGC. Here, we aimed to investigate the prevalence, the clinic-pathologic features, and the impact on treatment outcome of clinical molecular subtypes in a new-diagnosed Western cohort of AGC.

2023 - Comment on: Reconstructive techniques following low anterior resection for carcinoma of the rectum: meta-analysis [Articolo su rivista]
Manenti, A.; Caramaschi, S.; Farinetti, A.; Manco, G.; Bonetti, L. R.
abstract

2023 - Digital real-time microscopy of ex-vivo tissues: A novel strategy to control surgical accuracy [Articolo su rivista]
Sighinolfi, Maria Chiara; Cimadamore, Alessia; Cassani, Alessandra; Assumma, Simone; Sarchi, Luca; Filippi, Beatrice; Turri, Filippo; Reggiani Bonetti, Luca; Maiorana, Antonino; Eissa, Ahmed; Micali, Salvatore; Montironi, Rodolfo; Rocco, Bernardo
abstract

Introduction: Ex-vivo FCM is a novel digital optical technique that provides images of fresh tissues in a real-time fashion with magnification to subcellular details of a flattened unprocessed sample. Digital images are hematoxylin-eosin-like and can be shared and interpreted remotely. In urology, FCM has been successfully applied for prostate tissue interpretation, either during biopsy and radical prostatectomy. Possible applications of FCM may reflect those of frozen section analysis and can be extended to all fields in which the intra-operative microscopical control is advisable. Materials and methods: This is an investigative prospective case series that aims to explore FCM feasibility in novel surgical settings and provide a depiction of FCM digital images in those fields. The definite purpose is to check the accuracy of surgical specimen during the following interventions: (a) trans-urethral resection of bladder tumors, to confirm the presence of muscular layer; (b) biopsy of a retroperitoneal mass, to check for the location and quality of cores; (c) training in robotic radical prostatectomy, to control surgical margins after a nerve sparing performed by a trainee. To this aim, we collected FCM images during seven surgical procedures. FCM findings were compared to those from the final histopathological analysis and the agreement was assessed. Results: In all cases, FCM digital images were obtained in the OR. FCM was able to confirm the presence of muscular layer in TURB specimen, the presence of lymphomatous tissue, surgical margins at prostate specimen. FCM intra-operative interpretation was consistent with final histopathology in all cases. Conclusions: Ex vivo FCM may represent a novel approach to control the quality of specimens, likely to tailor surgical strategy in a real-time fashion. Moreover, digitalization represents a step toward the implementation of telepathology in clinical practice.

2023 - Endoscopic Submucosal Dissection for Subepithelial Tumor Treatment in the Upper Digestive Tract: A Western, Multicenter Study [Articolo su rivista]
Manta, Raffaele; Zito, Francesco Paolo; Pugliese, Francesco; Caruso, Angelo; Mangiafico, Santi; D’Alessandro, Alessandra; Castellani, Danilo; Germani, Ugo; Mutignani, Massimiliano; Conigliaro, Rita Luisa; Bonetti, Luca Reggiani; Matsuda, Takahisa; De Francesco, Vincenzo; Zullo, Angelo; Galloro, Giuseppe
abstract

Background/aims: Endoscopic submucosal dissection (ESD) has been proposed for removal of gastrointestinal subepithelial tumors (GI-SETs), but data are still scanty. This study aimed to report a case series from a western country. Patients and methods: Data of patients with upper GI-SETs suitable for ESD removal observed in 4 centers were retrospectively reviewed. Before endoscopic procedure, the lesion was characterized by endosonographic evaluation, histology, and CT scan. The en bloc resection and the R0 resection rates were calculated, as well as incidence of complications, and the 1-year follow-up was reported. Results: Data of 84 patients with esophageal (N = 13), gastric (N = 61), and duodenal (N = 10) GI-SETs were collected. The mean diameter of lesions was 26 mm (range: 12-110 mm). There were 17 gastrointestinal stromal tumors, 12 neuroendocrine tumors, 35 leiomyomas, 18 lipomas, and 2 hamartomas. En bloc and R0 resection were achieved in 83 (98.8%) and in 80 (95.2%) patients, respectively. Overall, a complication occurred in 11 (13.1%) patients, including bleeding (N = 7) and perforation (N = 4). Endoscopic approach was successful in all bleedings, but 1 patient who required radiological embolization, and in 2 perforations, while surgery was performed in the other patients. Overall, a surgical approach was eventually needed in 5 (5.9%), including 3 in whom R0 resection failed and 2 with perforation. Conclusions: Our study found that ESD may be an effective and safe alternative to surgical intervention for both benign and localized malignant GI-SETs.

2023 - Idiopathic pulmonary fibrosis and intestinal disorders: An observational study [Articolo su rivista]
Stefania, C.; Angela, B.; Stefania, C.; FABRETTI COSTANTINO, Antonio; Andrea, A. -S.; Gianrocco, M.; Tiziana, S.; Luca, R. B.
abstract

Background and aim: Idiopathic pulmonary fibrosis (IPF) is a chronic respiratory disease characterized by a progressive decline in lung function and a specific histopathologic pattern defined as usual interstitial pneumonia. Early diagnosis and new therapeutic protocols have contributed to a reduction in disease progression. Thus, some patients may develop extrapulmonary diseases including malignancies and chronic pathologies. The aim of this study was to investigate the frequency of intestinal disorders such as polyps, colorectal carcinoma (CRC), and chronic inflammatory bowel disease (IBD) in patients with IPF. Methods: From the database of 189 patients with IPF (148 males, 78.3 %; 41 females, 21.7 %) residing in the district of Modena, we identified 44 patients (36 males, 81.8 %; 8 females, 18.2 %) with a histologically confirmed intestinal disease. Results: Intestinal polyps were detected in 41 cases (93.2 %), of which 4 were associated with CRC and 1 with IBD; 1 patient had only CRC (2.3 %), and 2 patients had only IBD (4.5 %). Both males and females developed bowel disease, but males seemed to have a higher number of polyps and high-grade adenomas with a predisposition to malignant transformation. Conclusions: As patients with IPF may present with intestinal diseases that can evolve into malignancies in some cases, they should undergo appropriate follow-up and targeted colorectal screening. Thus, colorectal pathologies should not take a back seat. These preliminary results encourage further research to select suitable patients for specific diagnostic and therapeutic procedures in order to prolong survival and improve the quality of life.

2023 - Molecular portraits of patients with intrahepatic cholangiocarcinoma who diverge as rapid progressors or long survivors on chemotherapy [Articolo su rivista]
O'Rourke, Colm J; Salati, Massimiliano; Rae, Colin; Carpino, Guido; Leslie, Holly; Pea, Antonio; Prete, Maria G; Bonetti, Luca R; Amato, Francesco; Montal, Robert; Upstill-Goddard, Rosie; Nixon, Colin; Sanchon-Sanchez, Paula; Kunderfranco, Paolo; Sia, Daniela; Gaudio, Eugenio; Overi, Diletta; Cascinu, Stefano; Hogdall, Dan; Pugh, Sian; Domingo, Enric; Primrose, John N; Bridgewater, John; Spallanzani, Andrea; Gelsomino, Fabio; Llovet, Josep M; Calvisi, Diego F; Boulter, Luke; Caputo, Francesco; Lleo, Ana; Jamieson, Nigel B; Luppi, Gabriele; Dominici, Massimo; Andersen, Jesper B; Braconi, Chiara
abstract

Objective: Cytotoxic agents are the cornerstone of treatment for patients with advanced intrahepatic cholangiocarcinoma (iCCA), despite heterogeneous benefit. We hypothesised that the pretreatment molecular profiles of diagnostic biopsies can predict patient benefit from chemotherapy and define molecular bases of innate chemoresistance. Design: We identified a cohort of advanced iCCA patients with comparable baseline characteristics who diverged as extreme outliers on chemotherapy (survival <6 m in rapid progressors, RP; survival >23 m in long survivors, LS). Diagnostic biopsies were characterised by digital pathology, then subjected to whole-transcriptome profiling of bulk and geospatially macrodissected tissue regions. Spatial transcriptomics of tumour-infiltrating myeloid cells was performed using targeted digital spatial profiling (GeoMx). Transcriptome signatures were evaluated in multiple cohorts of resected cancers. Signatures were also characterised using in vitro cell lines, in vivo mouse models and single cell RNA-sequencing data. Results: Pretreatment transcriptome profiles differentiated patients who would become RPs or LSs on chemotherapy. Biologically, this signature originated from altered tumour-myeloid dynamics, implicating tumour-induced immune tolerogenicity with poor response to chemotherapy. The central role of the liver microenviroment was confrmed by the association of the RPLS transcriptome signature with clinical outcome in iCCA but not extrahepatic CCA, and in liver metastasis from colorectal cancer, but not in the matched primary bowel tumours. Conclusions: The RPLS signature could be a novel metric of chemotherapy outcome in iCCA. Further development and validation of this transcriptomic signature is warranted to develop precision chemotherapy strategies in these settings.

2023 - Research of Prostate Cancer Urinary Diagnostic Biomarkers by Proteomics: The Noteworthy Influence of Inflammation [Articolo su rivista]
Bellei, Elisa; Caramaschi, Stefania; Giannico, Giovanna A; Monari, Emanuela; Martorana, Eugenio; Reggiani Bonetti, Luca; Bergamini, Stefania
abstract

: Nowadays, in the case of suspected prostate cancer (PCa), tissue needle biopsy remains the benchmark for diagnosis despite its invasiveness and poor tolerability, as serum prostate-specific antigen (PSA) is limited by low specificity. The aim of this proteomic study was to identify new diagnostic biomarkers in urine, an easily and non-invasively available sample, able to selectively discriminate cancer from benign prostatic hyperplasia (BPH), evaluating whether the presence of inflammation may be a confounding parameter. The analysis was performed by two-dimensional gel electrophoresis (2-DE), mass spectrometry (LC-MS/MS) and Enzyme-Linked Immunosorbent Assay (ELISA) on urine samples from PCa and BPH patients, divided into subgroups based on the presence or absence of inflammation. Significant quantitative and qualitative differences were found in the urinary proteomic profile of PCa and BPH groups. Of the nine differentially expressed proteins, only five can properly be considered potential biomarkers of PCa able to discriminate the two diseases, as they were not affected by the inflammatory process. Therefore, the proteomic research of novel and reliable urinary biomarkers of PCa should be conducted considering the presence of inflammation as a realistic interfering element, as it could hinder the detection of important protein targets.

2023 - Revolutionizing Cancer Research: The Impact of Artificial Intelligence in Digital Biobanking [Articolo su rivista]
Frascarelli, C.; Bonizzi, G.; Musico, C. R.; Mane, E.; Cassi, C.; Guerini Rocco, E.; Farina, A.; Scarpa, A.; Lawlor, R.; Reggiani Bonetti, L.; Caramaschi, S.; Eccher, A.; Marletta, S.; Fusco, N.
abstract

Background: Biobanks are vital research infrastructures aiming to collect, process, store, and distribute biological specimens along with associated data in an organized and governed manner. Exploiting diverse datasets produced by the biobanks and the downstream research from various sources and integrating bioinformatics and "omics" data has proven instrumental in advancing research such as cancer research. Biobanks offer different types of biological samples matched with rich datasets comprising clinicopathologic information. As digital pathology and artificial intelligence (AI) have entered the precision medicine arena, biobanks are progressively transitioning from mere biorepositories to integrated computational databanks. Consequently, the application of AI and machine learning on these biobank datasets holds huge potential to profoundly impact cancer research. Methods: In this paper, we explore how AI and machine learning can respond to the digital evolution of biobanks with flexibility, solutions, and effective services. We look at the different data that ranges from specimen-related data, including digital images, patient health records and downstream genetic/genomic data and resulting "Big Data" and the analytic approaches used for analysis. Results: These cutting-edge technologies can address the challenges faced by translational and clinical research, enhancing their capabilities in data management, analysis, and interpretation. By leveraging AI, biobanks can unlock valuable insights from their vast repositories, enabling the identification of novel biomarkers, prediction of treatment responses, and ultimately facilitating the development of personalized cancer therapies. Conclusions: The integration of biobanking with AI has the potential not only to expand the current understanding of cancer biology but also to pave the way for more precise, patient-centric healthcare strategies.

2022 - Biallelic PMS2 Mutations in a Family with Uncommon Clinical and Molecular Features [Articolo su rivista]
Pedroni, Monica; Ponz de Leon, Maurizio; Reggiani Bonetti, Luca; Rossi, Giuseppina; Viel, Alessandra; Urso, Emanuele Damiano Luca; Roncucci, Luca
abstract

: We describe a patient with constitutional mismatch repair-deficiency (CMMR-D) in whom the syndrome started at age 10 with the development of multiple adenomas in the large bowel. In the successive 25 years, four malignancies developed in different organs (rectum, ileum, duodenum, and lymphoid tissue). The patient had biallelic constitutional pathogenic variants in the PMS2 gene. We speculate that besides the PMS2 genotype, alterations of other genes might have contributed to the development of the complex phenotype. In the nuclear family, both parents carried different PMS2 germline mutations. They appeared in good clinical condition and did not develop polyps or cancer. The index case had a brother who died at age three of lymphoblastic leukemia, and a sister who was affected by sarcoidosis. Tumor tissue showed diffuse DNA microsatellite instability. A complete absence of immunoreactivity was observed for the PMS2 protein both in the tumors and normal tissues. Next-generation sequencing and multiple ligation-dependent probe amplification analyses revealed biallelic PMS2 germline pathogenic variants in the proband (genotype c.[137G>T];[(2174+1_2175-1)_(*160_?)del]), and one of the two variants was present in both parents-c.137G>T in the father and c.(2174+1-2175-1)_(*160_?)del in the mother-as well as c.137G>T in the sister. Moreover, Class 3 variants of MSH2 (c.1787A>G), APC (c.1589T>C), and CHEK2 (c.331G>T) genes were also detected in the proband. In conclusion, the recognition of CMMR-D may sometimes be difficult; however, the possible role of constitutional alterations of other genes in the development of the full-blown phenotype should be investigated in more detail.

2022 - Colon cancer in a 12-year-old girl with hypertriglyceridemia [Articolo su rivista]
Pedroni, Monica; Leon, Maurizio Ponz de; Bonetti, Luca Reggiani; Viel, Alessandra; Noto, Davide; Nascimbeni, Fabio; Sena, Paola; Roncucci, Luca
abstract

2022 - CtDNA-guided rechallenge with anti-EGFR therapy in RASwt metastatic colorectal cancer: Evidence from clinical practice [Articolo su rivista]
D’Onofrio, Raffaella; Caputo, Francesco; Prampolini, Francesco; Spallanzani, Andrea; Gelsomino, Fabio; Bettelli, Stefania; Manfredini, Samantha; Reggiani Bonetti, Luca; Carotenuto, Pietro; Bocconi, Alessandro; Dominici, Massimo; Luppi, Gabriele; Salati, Massimiliano
abstract

Aim: To apply extended ctDNA-based RAS genotyping to clinical criteria for improving the selection of patients eligible for anti-EGFR-based rechallenge in a real-world setting. Methods: ctDNA testing was prospectively applied to RASwt mCRC progressed after a first-line anti-EGFR-containing regimen and at least one other line. The primary endpoint was the objective response rate. Results: Among ten enrolled patients, the anti-EGFR rechallenge resulted in an objective response rate and disease control rate of 70% and 90%. The median progression-free survival was 11.3 months and overall survival was not reached. Compared with a historical cohort retreated with anti-EGFR agents based on clinical criteria, the ctDNA-driven approach resulted in a higher chance of achieving an objective response and longer survival. Conclusions: Blood-based RASwt status may enrich metastatic colorectal cancer more likely to benefit from anti-EGFR-based rechallenge. RAS genotyping in ctDNA represents a feasible, fast, and cost-effective tool to be implemented in the clinic for advancing precision medicine.

2022 - Development and Multicentre Validation of the Modena Score to Predict Survival in Advanced Biliary Cancers Undergoing Second-Line Chemotherapy [Articolo su rivista]
Salati, M.; Marcheselli, L.; Messina, C.; Merz, V.; Messina, M.; Carotenuto, P.; Caputo, F.; Gelsomino, F.; Spallanzani, A.; Bonetti, L. R.; Caramaschi, S.; Luppi, G.; Dominici, M.; Ghidini, M.
abstract

Background: The role of second-line chemotherapy in advanced biliary cancers (ABCs) has only recently been established in phase III randomized trial and the optimal selection of patients most likely to benefit from it remains challenging. Methods: A cohort of 98 ABC treated second-line chemotherapy was used as a developmental dataset to identify covariates independently associated with overall survival (OS). Kaplan–Meier analysis was used to investigate the association between variables and OS and those retaining statistically significance were combined in a multiplexed score. Results: The following pretreatment variables were independently associated with OS: ECOG PS > 0, peritoneal disease, LDH > 430 UI/L, albumin <3.5 gr/dL, gamma-GT >100 UI/L, sodium <140 mEq/L, absolute lymphocyte count <1000/mmc, and PFS to first-line <6 months. Based on these results, a scoring system was developed that identified three subgroups with statistically different OS: low-risk (mOS 18 months), intermediate-risk (mOS 9.4 months) and high-risk (mOS 2.9 months) (p < 0.001). The prognostic model was both internally and externally validated in a multicentre cohort of 120 ABCs. Conclusion: The Modena score is a multiplexed scoring system capable of accurately risk-stratified ABCs treated with second-line chemotherapy. Based on its reproducibility, usability and generalizability, it has the potential for assisting therapeutic decision-making in the clinic and risk-stratification in future trials.

2022 - Eczema: Features on FCM, Digital H&E, and Corresponding Conventional H&E [Capitolo/Saggio]
Bertoni, L.; Reggiani, C.; Azzoni, P.; Reggiani, Bonetti; Pellacani, G
abstract

2022 - Effects of Energy Drink Acute Assumption in Gastrointestinal Tract of Rats [Articolo su rivista]
Nasi, Milena; De Gaetano, Anna; Carnevale, Gianluca; Bertoni, Laura; Selleri, Valentina; Zanini, Giada; Pisciotta, Alessandra; Caramaschi, Stefania; Reggiani Bonetti, Luca; Farinetti, Alberto; Cossarizza, Andrea; Pinti, Marcello; Manenti, Antonio; Mattioli, Anna Vittoria
abstract

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2022 - Expression of Autophagic and Inflammatory Markers in Normal Mucosa of Individuals with Colorectal Adenomas: A Cross Sectional Study among Italian Outpatients Undergoing Colonoscopy [Articolo su rivista]
Sena, Paola; Mancini, Stefano; Pedroni, Monica; Reggiani Bonetti, Luca; Carnevale, Gianluca; Roncucci, Luca
abstract

: Colorectal cancer (CRC) ranks among the three most common cancers in terms of both cancer incidence and cancer-related deaths in Western industrialized countries. Lifetime risk of colorectal cancer may reach 6% of the population living in developed countries. In the current era of personalized medicine, CRC is no longer considered as a single entity. In more recent years many studies have described the distinct differences in epidemiology, pathogenesis, genetic and epigenetic alterations, molecular pathways and outcome depending on the anatomical site. The aim of our study is to assess in a multidimensional model the association between metabolic status and inflammatory and autophagic changes in the normal colorectal mucosa classified as right-sided, left-sided and rectum, and the presence of adenomas. One hundred and sixteen patients undergoing colonoscopy were recruited and underwent a complete serum lipid profile, immunofluorescence analysis of colonic biopsies for MAPLC3 and myeloperoxidase expression, matched with clinical and anthropometric characteristics. Presence of adenomas correlated with cholesterol (total and LDL) levels, IL-6 levels, and MAPLC3 tissue expression, especially in the right colon. In conclusion, serum IL-6 amount and autophagic markers could be good predictors of the presence of colorectal adenomas.

2022 - Features of Lichen Planus and Psoriasis on Ex Vivo Confocal Imaging and Histopathologic Correlation [Capitolo/Saggio]
Bertoni, L.; Reggiani, C.; Azzoni, P.; Bonetti, L. R.; Pellacani, G
abstract

2022 - Fluorescence Confocal Microscope for Identification of Discoid Lupus Erythematosus Histologic Features [Capitolo/Saggio]
Bertoni, L.; Reggiani, C.; Azzoni, P.; Caramaschi, S.; Bonetti, L. R.; Pellacani, G
abstract

2022 - Leg-type cutaneous B-cell lymphoma: The description of a rare and aggressive skin tumor with D-OCT imaging [Articolo su rivista]
Chello, C.; Caramaschi, S.; Sticchi, A.; Naselli, A.; Pezzini, C.; Farnetani, F.; Reggiani Bonetti, L.; Ciardo, S.; Di Matteo, E.; Pellacani, G.; Manfredini, M.
abstract

2022 - New digital confocal laser microscopy may boost real-time evaluation of endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) from solid pancreatic lesions: Data from an international multicenter study [Articolo su rivista]
Amendoeira, Isabel; Arcidiacono, Paolo Giorgio; Barizzi, Jessica; Capitanio, Arrigo; Cuatrecasas, Miriam; Di Matteo, Francesco Maria; Doglioni, Claudio; Fukushima, Noriyoshi; Fulciniti, Franco; Ginès, Angels; Giovannini, Marc; Zaibo, Li; Lopes, Joanne; Lujan, Giovanni; Parisi, Alice; Poizat, Flora; Reggiani Bonetti, Luca; Stigliano, Serena; Taffon, Chiara; Verri, Martina; Crescenzi, Anna
abstract

Background: Pancreatic cancer is an aggressive malignancy and a leading cause of cancer death worldwide; its lethality is partly linked to the difficulty of early diagnosis. Modern devices for endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) were recently developed to improve targeting and sampling of small lesions, but innovative technologies for microscopic assessment are still lacking. Ex vivo fluorescence confocal laser microscopy (FCM) is a new digital tool for real-time microscopic assessment of fresh unfixed biological specimens, avoiding conventional histological slide preparation and potentially being highly appealing for EUS-FNB specimens. Methods: This study evaluated the possible role of FCM for immediate evaluation of pancreatic specimens from EUS-FNB. It involved comparison of the interobserver agreement between the new method and standard histological analysis during international multicenter sharing of digital images. Digital images from 25 cases of EUS-FNB obtained with real-time FCM technology and 25 paired digital whole-slide images from permanent conventional paraffin sections were observed by 10 pathologists from different Institutions in Europe, Japan, and the United States, in a blinded manner. The study evaluated 500 observations regarding adequacy, morphological clues, diagnostic categories, and final diagnosis. Findings: Statistical analysis showed substantial equivalence in the interobserver agreement among pathologists using the two techniques. There was also good inter-test agreement in determining sample adequacy and when assigning a diagnostic category. Among morphological features, nuclear enlargement was the most reproducible clue, with very good inter-test agreement. Interpretation: Findings in this study are from international multicenter digital sharing and are published here for the first time. Considering the advantages of FCM digital diagnostics in terms of reduced time and unaltered sample maintenance, the ex vivo confocal laser microscopy may effectively improve traditional EUS-FNB diagnostics, with significant implications for planning modern diagnostic workflow for pancreatic tumors. Funding: This study was not supported by any funding source.

2022 - Preclinical Validation of a Semi-Autonomous Robot for Transperineal Prostate Biopsy [Articolo su rivista]
Maris, B.; Fiazza, M. -C.; De Piccoli, M.; Tenga, C.; Palladino, L.; Puliatti, S.; Iseppi, A.; Ferrari, R.; Piro, A.; Bonetti, L. R.; Ligabue, G.; Tafuri, A.; Micali, S.; Fiorini, P.
abstract

Prostate biopsy is a manual procedure carried out mostly under ultrasound (US) guidance to confirm the presence of cancer. The standard biopsy is random and includes at least 12 insertions; targeted biopsy makes use of dedicated hardware and software, but is still performed manually. We present here the pre-clinical validation of PROST, a robot primarily designed to automate targeted transperineal biopsy. The overall validation of the system was performed on cadavers, while some features, such as image segmentation, were tested on human tissue. PROST is designed to minimize human error by introducing some autonomy in the execution of key steps of the procedure, i.e., target selection, image fusion and needle positioning. The protocol was approved by the ethics committee; 10 cadavers were included in the study. We envision that PROST has the potential to increase the detection of clinically significant prostate cancer, to simplify the procedure, to reduce human errors and to shorten training time. The use of a robot for the biopsy of the prostate will create the possibility to include also a treatment, such as focal ablation, to be delivered through the same system.

2022 - Role of poorly differentiated cluster in gastric cancer: is it a new prognosis factor? [Articolo su rivista]
Sorrentino, L; De Ruvo, N; Serra, F; Salati, M; Ricciardolo, A A; Bonetti, L R; Gelmini, R
abstract

Poorly differentiated Clusters (PDCs) of tumor cells composed of more than five elements have been recently described in gastrointestinal cancers and correlate with a worse prognosis. Our study aims to investigate PDC occurrence in a series of patients with gastric cancer and correlate it with lymph node status and clinical outcome.

2022 - Statins increase pathological response in locally advanced rectal cancer treated with chemoradiation: a multicenter experience. [Articolo su rivista]
Caputo, F; Santini, C; Casadei-Gardini, A; Cerma, K; Bardasi, C; Garajovà, I; Lattanzi, E; Passardi, A; Rapposelli, Ig; Spallanzani, A; Salati, M; Bonetti, Lr; Gelmini, R; Meduri, B; Piccoli, M; Pecchi, A; Benatti, S; Piacentini, F; Dominici, M; Luppi, G; Gelsomino, F.
abstract

Aims: To investigate the influence of various concomitant medications on outcomes in patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiation. Materials & methods: The authors retrospectively identified 246 patients from 2003 to 2018, collecting demographic and clinicopathological data of interest. Odds ratio (OR) was used to assess the association between concomitant drugs and outcomes. Results: The authors found an association between statins and a Dworak regression grade of 3-4 (OR = 8.78; p = 0.01). Furthermore, statins were significantly associated with more frequent chemoradiation-related toxicity (OR = 2.39; p = 0.0098) and chemotherapy dose reduction or discontinuation (OR = 2.26; p = 0.03). Conclusion: Despite higher frequency of radiotherapy and chemotherapy interruption or dose reduction, the concomitant use of statins during neoadjuvant chemoradiation proved to be associated with better tumor regression.

2021 - Alternative splicing of NF-YA promotes prostate cancer aggressiveness and represents a new molecular marker for clinical stratification of patients [Articolo su rivista]
Belluti, Silvia; Semeghini, Valentina; Rigillo, Giovanna; Ronzio, Mirko; Benati, Daniela; Torricelli, Federica; Reggiani Bonetti, Luca; Carnevale, Gianluca; Grisendi, Giulia; Ciarrocchi, Alessia; Dominici, Massimo; Recchia, Alessandra; Dolfini, Diletta; Imbriano, Carol
abstract

Approaches based on expression signatures of prostate cancer (PCa) have been proposed to predict patient outcomes and response to treatments. The transcription factor NF-Y participates to the progression from benign epithelium to both localized and metastatic PCa and is associated with aggressive transcriptional profile. The gene encoding for NF-YA, the DNA-binding subunit of NF-Y, produces two alternatively spliced transcripts, NF-YAs and NF-YAl. Bioinformatic analyses pointed at NF-YA splicing as a key transcriptional signature to discriminate between different tumor molecular subtypes. In this study, we aimed to determine the pathophysiological role of NF-YA splice variants in PCa and their association with aggressive subtypes.

2021 - Alternative splicing of NF-YA promotes prostate cancer aggressiveness and represents a new molecular marker for clinical stratification of patients [Poster]
Belluti, Silvia; Semeghini, Valentina; Rigillo, Giovanna; Ronzio, Mirko; Benati, Daniela; Torricelli, Federica; REGGIANI BONETTI, Luca; Carnevale, Gianluca; Grisendi, Giulia; Ciarrocchi, Alessia; Dominici, Massimo; Recchia, Alessandra; Dolfini, Diletta; Imbriano, Carol
abstract

2021 - Current and future perspectives of digital microscopy with fluorescence confocal microscope for prostate tissue interpretation: A narrative review [Articolo su rivista]
Rocco, B.; Cimadamore, A.; Sarchi, L.; Bonetti, L. R.; Bertoni, L.; Azzoni, P.; Assumma, S.; Turri, F.; Bozzini, G.; Eissa, A.; Micali, S.; Bianchi, G.; Maiorana, A.; Montironi, R.; Pellacani, G.; Sighinolfi, M. C.
abstract

Fluorescence confocal microscopy (FCM) is an optical imaging technique providing digital microscopical images of fresh tissue in a real time fashion, without conventional processing. FCM has been widely applied in several fields of dermatology, including the detection of basal cell carcinoma and of cutaneous inflammatory diseases. The aim of the paper is to provide an overview of FCM applications in the field of prostate tissue interpretation and prostate cancer (PCa) detection. A Literature search (PubMed & Web of Science) was performed to identify articles concerned with the clinical and surgical applications of FCM in prostatic and periprostatic tissues interpretation. Overall, six articles were identified. All articles investigated the level of agreement between FCM and conventional histopathological analysis (hematoxylin-eosin, HE) for the discrimination between normal and PCa tissues. An investigative article on prostate samples retrieved from radical prostatectomy (RP) specimens and an atlas of FCM digital images from the same series were found. Two prospective clinical trials, comparing FCM and HE, pointed out a “substantial” to “almost perfect” discriminative performance of FCM for the diagnosis of PCa on prostate biopsy core. Finally, two studies investigated the intra-operative role of FCM during RP for the control of surgical dissection. In this setting, FCM could be used to analyse samples retrieved from suspicious peri-prostatic areas; FCM has also been tested for an en-face evaluation of flat slices obtained from the systematic sampling of the posterolateral aspects of the prostate, in a NeuroSAFE-like approach. Generally, FCM provides digital microscopical images of fresh tissue in a real time fashion, without requiring conventional processing. Currently, available studies confirmed a high concordance with conventional pathology for the detection of PCa. Further studies are required to validate the technology, to evaluate ISUP score attribution and to implement the fields of application of FCM for the treatment of prostate diseases.

2021 - Cutaneous metastasis from colorectal cancer: Making light on an unusual and misdiagnosed event [Articolo su rivista]
Parente, P.; Ciardiello, D.; Reggiani Bonetti, L.; Famiglietti, V.; Cazzato, G.; Caramaschi, S.; Attino, V.; Urbano, D.; Di Maggio, G.; Ingravallo, G.
abstract

Cutaneous metastasis from solid tumors is a rare event and usually represents a late occurrence in the natural history of an advanced visceral malignancy. Rarely, cutaneous metastasis has been described in colorectal cancer patients. The most frequent cutaneous site of colorectal metastasis is the surgical scar in the abdomen following the removal of the primary malignancy, followed by the extremities, perineum, head, neck, and penis. Metastases to the thigh and back of the trunk are anecdotical. Dermatological diagnosis of cutaneous metastasis can be quite complex, especially in unusual sites, such as in the facial skin or thorax and in cases of single cutaneous lesions since metastasis from colorectal cancer is not usually the first clinical hypothesis, leading to misdiagnosis. To date, due to the rarity of cutaneous metastasis from colorectal cancer, little evidence, most of which is based on case reports and very small case series, is currently available. Therefore, a better understanding of the clinic-pathological characteristics of this unusual metastatic site represents an unmet clinical need. We present a large series of 29 cutaneous metastases from colorectal cancer with particular concerns regarding anatomic localization and the time of onset with respect to primitive colorectal cancer and visceral metastases.

2021 - Cytoproliferative activity in colorectal poorly differentiated clusters: Biological significance in tumor setting [Articolo su rivista]
Caramaschi, S.; Mangogna, A.; Salviato, T.; Ammendola, S.; Barresi, V.; Manco, G.; Canu, P. G.; Zanelli, G.; Bonetti, L. R.
abstract

Poorly differentiated clusters (PDCs) have gained a significant prognostic role in colorectal carcinomas (CRCs) being associated to high risk of lymph node metastasis, shorter survival time and poor prognosis. The knowledge in PDC biology is not completely clear.

2021 - Diagnostic interobserver variability in Crohn's disease- and ulcerative colitis-associated dysplasia: a multicenter digital survey from the IG-IBD Pathologists Group [Articolo su rivista]
Leoncini, G.; Donato, F.; Reggiani Bonetti, L.; Salviato, T.; Cadei, M.; Daperno, M.; Principi, M. B.; Armuzzi, A.; Caprioli, F.; Canavese, G.; Villanacci, V.
abstract

BACKGROUND: Crohn's disease (CD) and ulcerative colitis, two forms of inflammatory bowel disease (IBD), are chronic and relapsing conditions of the gastrointestinal tract both characterized by long lasting chronic inflammation and increased risk of dysplasia and colorectal cancer (CRC). The aim of our study was to evaluate the interobserver agreement about IBD-associated dysplasia among pathologists belonging to the Italian Group for Inflammatory Bowel Diseases (IG-IBD P). METHODS: The present multicenter survey was performed using telepathology, supported by an open source E-learning platform. Biopsy specimens from 30 colonoscopies and from 20 patients were included. The glass slides of any case, including clinical and endoscopic data, were digitalized and uploaded on the E-learning platform. All the digital slides were grouped in 54 diagnostic "blocks". Blinded histopathological evaluation on all the digital slides was performed by 20 gastrointestinal pathologists. Closed-ended questions about (1) the occurrence of IBD; (2) the classification of IBD (as UC or CD); (3) the presence of active versus quiescent disease; (4) the presence of dysplasia; (5) the possible association of dysplasia with the sites of disease (dysplasia-associated lesion or mass-DALM vs adenoma-like mass-ALM); (6) the grading of dysplasia according to the ECCO guidelines (negative, indefinite, low grade, high grade categories) and (7) the presence of associated serrated features, were proposed in each case. Inter-observer agreement was evaluated by mean agreement percentage and kappa statistic, when suitable. RESULTS: The diagnosis of IBD was confirmed in 19 of 20 patients, 17 of 19 being classified as UC, 2 as CD. The mean interobserver agreement percentages about (1) the evidence of IBD, (2) the presence of either UC or CD and (3) the activity grading resulted to be 80%, 69% and 86%, respectively. Dysplasia was detected in 8/20 patients, with moderate agreement between pathologists (mean 72%, k 0.48). Particularly, low grade dysplasia was found in 13 biopsies (combined k 0.38), whereas high grade dysplasia in 8 (combined k 0.47). When the endoscopic and histopathological data were combined, features consistent with DALM were found in 6 of 20 patients with low grade dysplasia and those consistent with ALM in 2 patients with low grade dysplasia in a single biopsy (mean agreement: 86%). An associated serrated pattern was discovered in 4 patients (7 biopsies). CONCLUSIONS: Our study showed moderate interobserver agreement about the histopathological detection and classification of IBD-associated dysplasia. Further efforts should be undertaken to integrate the histopathological data with both the ancillary tests and molecular investigations.

2021 - Digital Frozen Sections with Fluorescence Confocal Microscopy During Robot-assisted Radical Prostatectomy: Surgical Technique [Articolo su rivista]
Rocco, B.; Sarchi, L.; Assumma, S.; Cimadamore, A.; Montironi, R.; Reggiani Bonetti, L.; Turri, F.; De Carne, C.; Puliatti, S.; Maiorana, A.; Pellacani, G.; Micali, S.; Bianchi, G.; Sighinolfi, M. C.
abstract

Background: Robot-assisted radical prostatectomy (RARP) involves a tradeoff between oncological control and functional outcomes. Intraoperative control of surgical margins (SMs) may help in ensuring the safety of the dissection. Fluorescence confocal microscopy (FCM) is an effective method for interpretation of prostate tissue and provides digital images with an appearance similar to hematoxylin-eosin staining. Objective: To describe an alternative technique to NeuroSAFE for intraoperative evaluation of neurovascular-adjacent margins shaved from ex vivo specimens using FCM analysis. Design, setting, and participants: This was a prospective study of 24 patients undergoing RARP with intraoperative FCM control of margin status. Surgical procedure: After surgical dissection, SMs are sectioned from the fresh prostate using the Mohs technique (shaving): three slices from the apex and the right and left posterolateral aspects are obtained. Digital images of the shavings are immediately acquired via FCM and shared with a remote pathologist. In the case of a positive SM, a focal secondary resection of the bundle can be performed owing to the ability of FCM to locate a region of interest on the flat sample. Measurements: The primary outcome measure was the rate of negative margins at neurovascular-adjacent sites. Oncological and functional outcomes for patients with 1 yr of follow-up are also reported. Results and limitations: All patients had negative SMs in shavings from neurovascular-adjacent areas at final histopathology; four underwent a secondary resection with final conversion to negative SM status. Nine of ten patients with 1-yr follow-up are free of biochemical recurrence (prostate-specific antigen persistence in one pN1 case), nine are fully continent, and four of the five with preoperative potency have recovered their sexual function. Conclusions: Digital frozen sections with FCM during RARP may represent an alternative to NeuroSAFE for possible optimization of functional outcomes without compromising oncological safety. Patient summary: We developed a technique to ensure complete removal of cancer tissue during surgical removal of the prostate. Tissue specimens are examined via digital microscopy in real time during the operation. This allows the surgeon to remove more tissue if cancer is detected at the margins of a specimen, while avoiding unnecessary removal of healthy tissue.

2021 - Endoscopic submucosal dissection for visible dysplasia treatment in ulcerative colitis patients: Cases series and systematic review of literature [Articolo su rivista]
Manta, R.; Zullo, A.; Telesca, D. A.; Castellani, D.; Germani, U.; Reggiani Bonetti, L.; Conigliaro, R.; Galloro, G.
abstract

Background and Aims: Ulcerative colitis [UC] patients are at an increased risk of developing colorectal cancer due to chronic inflammation. Endoscopic submucosal dissection [ESD] allows removal of non-invasive neoplastic lesions in the colon, but few data are available on its efficacy in UC patients. Methods: Data from consecutive UC patients diagnosed with visible dysplastic lesions in the colon who underwent ESD were evaluated. The en bloc removal, R0 resection and complication rates were calculated. Local recurrence and metachronous lesions during follow-up were identified. A systematic review of the literature with pooled data analysis was performed. Results: A total of 53 UC patients [age: 65 years; range 30-74; M/F: 31/22] underwent ESD. The en bloc resection rate was 100%, and the R0 resection rate was 96.2%. Bleeding occurred in seven [13.2%] patients, and perforation in three [5.6%] cases, all treated at endoscopy. No recurrence was observed, but two metachronous lesions were detected. Data from six other studies [three Asian and three European] were available. By pooling data, en bloc resection was successful in 88.4% (95% confidence interval [CI] = 83.5-92) of 216 lesions and in 91.8% [95% CI = 87.3-94.8] of 208 patients. R0 resection was achieved in 169 ESDs, equivalent to a 78.2% [95% CI = 72.3-83.2] rate for lesions and 81.3% [95% CI = 75.4-86] rate for patients. No difference between European and Asian series was noted. Conclusions: This pooled data analysis indicated that ESD is a suitable tool for safely and properly removing non-invasive neoplastic lesions on colonic mucosa of selected UC patients.

2021 - Histopathology of IBD colitis. A practical approach from the pathologists of the Italian group for the study of the gastrointestinal tract (GIPAD) [Articolo su rivista]
Villanacci, V.; Reggiani Bonetti, L.; Salviato, T.; Leoncini, G.; Cadei, M.; Albarello, L.; Caputo, A.; Aquilano, M. C.; Battista, S.; Parente, P.
abstract

Inflammatory bowel diseases (IBDs) are lifelong disorders in which an interaction between genetic and environmental factors is involved. IBDs include two entities: Crohn’s disease (CD) and ulcerative colitis (UC); these can be adequately diagnosed and distinguished with a correct methodological approach based on communicating exhaustive clinical, endoscopic and laboratory information to the pathologist and performing adequate bioptic sampling and precise morphological signs including crypt architecture, distribution of inflammation and granulomas, when present. IBD needs to be distinguished from non-IBD colitis, mostly at its onset. Moreover, IBDs are associated with an increased risk of developing colorectal adenocarcinoma. In daily pathological practice, correct diagnosis of IBD and its subclassification as well as a correct detection of dysplasia is imperative to establish the best therapeutic approach.

2021 - Histopathology of non-IBD Colitis. A practical approach from the Italian group for the study of the gastrointestinal tract (GIPAD) [Articolo su rivista]
Villanacci, V.; Reggiani Bonetti, L.; Leoncini, G.; Parente, P.; Cadei, M.; Albarello, L.; Mandelli, G.; Caputo, A.
abstract

Non-IBD colitides (NIBDC) are intestinal diseases clinically and endoscopically overlapping with Inflammatory Bowel Diseases (IBD), sometimes with a similar histological picture. NIBDC include entities such as infectious colitis, ischemic colitis, pseudomembranous colitis, eosinophilic colitis, autoimmune enterocolitis, segmental colitis associated with diverticulosis, drug-induced colitis, radiation-induced colitis, diversion colitis, and microscopic colitis, this last including two entities: collagenous and lymphocytic colitis. The knowledge of the most useful histological features and the main clinical data for each entity is mandatory in daily clinical practice, for correct pathological diagnosis and clinical management.

2021 - Histopathology of non-IBD colitis practical recommendations from pathologists of IG-IBD Group [Articolo su rivista]
Reggiani Bonetti, L.; Leoncini, G.; Daperno, M.; Principi, M. B.; Baronchelli, C.; Manenti, S.; Caprioli, F.; Armuzzi, A.; Caputo, A.; Parente, P.; Cadei, M.; Villanacci, V.
abstract

Pathologists are often called upon to diagnose colitides that differ from the two main forms of inflammatory bowel disease (IBD). These non-IBD colitides include infectious colitis, microscopic colitis, ischemic colitis, eosinophilic colitis, autoimmune enterocolitis, segmental colitis associated with diverticulosis, drug-induced colitis, radiation colitis and diversion colitis. The diagnosis of these different disease entities relies on the histopathological examination of endoscopic biopsies of the gastrointestinal tract. This paper reviews the main histomorphological characteristics of the various Non-IBD colitides.

2021 - Identification of Sclerostin as a Putative New Myokine Involved in the Muscle-to-Bone Crosstalk [Articolo su rivista]
Magarò, Maria Sara; Bertacchini, Jessika; Florio, Francesca; Zavatti, Manuela; Potì, Francesco; Cavani, Francesco; Amore, Emanuela; De Santis, Ilaria; Bevilacqua, Alessandro; Reggiani Bonetti, Luca; Torricelli, Pietro; Maurel, Delphine B.; Biressi, Stefano; Palumbo, Carla
abstract

Bone and muscle have been recognized as endocrine organs since they produce and secrete “hormone-like factors” that can mutually influence each other and other tissues, giving rise to a “bone–muscle crosstalk”. In our study, we made use of myogenic (C2C12 cells) and osteogenic (2T3 cells) cell lines to investigate the effects of muscle cell-produced factors on the maturation process of osteoblasts. We found that the myogenic medium has inhibitory effects on bone cell differentiation and we identified sclerostin as one of the myokines produced by muscle cells. Sclerostin is a secreted glycoprotein reportedly expressed by bone/cartilage cells and is considered a negative regulator of bone growth due to its role as an antagonist of the Wnt/β-catenin pathway. Given the inhibitory role of sclerostin in bone, we analyzed its expression by muscle cells and how it affects bone formation and homeostasis. Firstly, we characterized and quantified sclerostin synthesis by a myoblast cell line (C2C12) and by murine primary muscle cells by Western blotting, real-time PCR, immunofluorescence, and ELISA assay. Next, we investigated in vivo production of sclerostin in distinct muscle groups with different metabolic and mechanical loading characteristics. This analysis was done in mice of different ages (6 weeks, 5 and 18 months after birth) and revealed that sclerostin expression is dynamically modulated in a muscle-specific way during the lifespan. Finally, we transiently expressed sclerostin in the hind limb muscles of young mice (2 weeks of age) via in vivo electro-transfer of a plasmid containing the SOST gene in order to investigate the effects of muscle-specific overproduction of the protein. Our data disclosed an inhibitory role of the muscular sclerostin on the bones adjacent to the electroporated muscles. This observation suggests that sclerostin released by skeletal muscle might synergistically interact with osseous sclerostin and potentiate negative regulation of osteogenesis possibly by acting in a paracrine/local fashion. Our data point out a role for muscle as a new source of sclerostin.

2021 - Prognostic Role of Mismatch Repair Status, Histotype and High-Risk Pathologic Features in Stage II Small Bowel Adenocarcinomas [Articolo su rivista]
Vanoli, A.; Grillo, F.; Guerini, C.; Neri, G.; Arpa, G.; Klersy, C.; Nesi, G.; Giuffrida, P.; Sampietro, G.; Ardizzone, S.; Fociani, P.; Fiocca, R.; Latella, G.; Sessa, F.; D'Errico, A.; Malvi, D.; Mescoli, C.; Rugge, M.; Ferrero, S.; Poggioli, G.; Rizzello, F.; Macciomei, M. C.; Santini, D.; Volta, U.; De Giorgio, R.; Caio, G.; Calabro, A.; Ciacci, C.; D'Armiento, M.; Rizzo, A.; Solina, G.; Martino, M.; Tonelli, F.; Villanacci, V.; Cannizzaro, R.; Canzonieri, V.; Florena, A. M.; Biancone, L.; Monteleone, G.; Caronna, R.; Ciardi, A.; Elli, L.; Caprioli, F.; Vecchi, M.; D'Inca, R.; Zingone, F.; D'Odorico, A.; Lenti, M. V.; Oreggia, B.; Reggiani Bonetti, L.; Giannone, A. G.; Orlandi, A.; Barresi, V.; Ciccocioppo, R.; Amodeo, G.; Biletta, E.; Luinetti, O.; Pedrazzoli, P.; Pietrabissa, A.; Corazza, G. R.; Solcia, E.; Paulli, M.; Di Sabatino, A.
abstract

Background: Small bowel adenocarcinoma is a relatively rare cancer, often diagnosed in an advanced stage. In localized and resectable disease, surgery alone or in combination with adjuvant chemotherapy is the mainstay of treatment. In the recently published National Comprehensive Cancer Network Clinical Practice guidelines, criteria for selecting patients with stage II small bowel adenocarcinoma to receive adjuvant chemotherapy are provided, and they are mainly extrapolated from studies on colorectal cancer. Patients and Methods: In the present study, we aimed to verify whether mismatch repair deficiency phenotype, high-risk pathologic features (including T4, positive resection margins and a low number of lymph nodes harvested), as well as tumor histologic subtype, were associated with cancer-specific survival in 66 stage II non-ampullary small bowel adenocarcinoma patients, collected through the Small Bowel Cancer Italian Consortium. A central histopathology review was performed. Mismatch repair deficiency was tested by immunohistochemistry for MLH1, MSH2, MSH6 and PMS2, and confirmed by polymerase chain reaction for microsatellite instability. Results: We identified mismatch repair deficiency, glandular/medullary histologic subtype, and celiac disease as significant predictors of favorable cancer-specific survival using univariable analysis with retained significance in bivariable models adjusted for pT stage. Among the high-risk features, only T4 showed a significant association with an increased risk of death; however, its prognostic value was not independent of mismatch repair status. Conclusions: Mismatch repair protein expression, histologic subtype, association with celiac disease, and, in the mismatch repair proficient subset only, T stage, may help identify patients who may benefit from adjuvant chemotherapy. Graphic Abstract: [Figure not available: see fulltext.]

2021 - TRAIL receptors are expressed in both malignant and stromal cells in pancreatic ductal adenocarcinoma [Articolo su rivista]
Dall'Ora, Massimiliano; Rovesti, Giulia; Reggiani Bonetti, Luca; Casari, Giulia; Banchelli, Federico; Fabbiani, Luca; Veronesi, Elena; Petrachi, Tiziana; Magistri, Paolo; Di Benedetto, Fabrizio; Spallanzani, Andrea; Chiavelli, Chiara; Spano, Maria Carlotta; Maiorana, Antonino; Dominici, Massimo; Grisendi, Giulia
abstract

: This study assesses the expression of all TNF-related apoptosis-inducing ligand (TRAIL) receptors in pancreatic ductal adenocarcinoma (PDAC) tumor tissue. We aimed to include TRAIL receptor expression as an inclusion parameter in a future clinical study using a TRAIL-based therapy approach for PDAC patients. Considering the emerging influence of PDAC desmoplastic stroma on the efficacy of anti-PDAC therapies, this analysis was extended to tumor stromal cells. Additionally, we performed PDAC stroma characterization. Our retrospective cohort study (N=50) included patients with histologically confirmed PDAC who underwent surgery. The expression of TRAIL receptors (DR4, DR5, DcR1, DcR2, and OPG) in tumor and stromal cells was evaluated by immunohistochemistry (IHC). The amount of tumor stroma was assessed by anti-vimentin IHC and Mallory's trichrome staining. The prognostic impact was determined by the univariate Cox proportional hazards regression model. An extensive expression of functional receptors DR4 and DR5 and a variable expression of decoy receptors were detected in PDAC tumor and stromal cells. Functional receptors were detected also in metastatic tumor and stromal cells. A poor prognosis was associated with low or absent expression of decoy receptors in tumor cells of primary PDAC. After assessing that almost 80% of tumor mass was composed of stroma, we correlated a cellular-dense stroma in primary PDAC with reduced relapse-free survival. We demonstrated that TRAIL functional receptors are widely expressed in PDAC, representing a promising target for TRAIL-based therapies. Further, we demonstrated that a low expression of DcR1 and the absence of OPG in tumor cells, as well as a cellular-dense tumor stroma, could negatively impact the prognosis of PDAC patients.

2021 - The Interplay between HGF/c-met Axis and Nox4 in BRAF Mutated Melanoma [Articolo su rivista]
Beretti, Francesca; Farnetani, Francesca; Reggiani Bonetti, Luca; Fabbiani, Luca; Zavatti, Manuela; Maiorana, Antonino; Pellacani, Giovanni; Maraldi, Tullia
abstract

Melanoma is the leading cause of death due to cutaneous malignancy and its incidence is on the rise. Several signaling pathways, including receptor tyrosine kinases, have a role in the development and progression of melanocytic lesions and malignant melanoma. Among those, the hepatocyte growth factor (HGF)/c-met axis is emerging as a critical player because it can play a role in drug resistance. Indeed, 50% of melanoma patients present BRAF mutations, however, all responders develop resistance to the inhibitors typically within one year of treatment. Interestingly, BRAF inhibitors induce reactive oxygen species (ROS) in melanoma cells, therefore, the aim of this study was to investigate a possible interplay between HGF/c-met and ROS sources, such as NADPH oxidases (Nox).

2021 - The evolving role of fgfr2 inhibitors in intrahepatic cholangiocarcinoma: From molecular biology to clinical targeting [Articolo su rivista]
Salati, M.; Caputo, F.; Baldessari, C.; Carotenuto, P.; Messina, M.; Caramaschi, S.; Dominici, M.; Bonetti, L. R.
abstract

Intrahepatic cholangiocarcinoma (iCCA) is an anatomically and biologically distinct entity with a rising incidence and a poor prognosis on conventional treatments. Surgery followed by adjuvant chemotherapy is a potentially curative option in resectable cases, while palliative-intent chemotherapy is the standard-of-care in the advanced setting. Technological advances through massive parallel sequencing have enabled a deeper under-standing of disease biology with the identification of several druggable molecular vulnerabilities in nearly 50% of cases. Among them, gene fusions involving the fibroblast growth factor receptor 2 (FGFR2) are the most therapeutically exploited so far with a number of Phase II clinical trials investigating FGFR2 inhibitors showing unprecedented efficacy results in this molecular subgroup. Over the last year, these efforts have culminated in the US FDA-approval of pemigatinib and infigratinib, the first two oral selective FGFR2 targeted agents for previously treated, locally advanced or metastatic iCCA driven by FGFR2 fusion or rearrangements. While first-line Phase III trials are currently underway to test these targeted approach against standard-of-care chemotherapy, translational studies are trying to better understand primary and secondary resistance mechanisms in order to optimize FGFR2 blockade in iCCA. In this article, we extensively reviewed the current evidence on the biological rationale, as well as preclinical and clinical development of FGFR inhibitors in iCCA.

2020 - An intraoperative study with ex vivo fluorescence confocal microscopy: diagnostic accuracy of the three visualization modalities [Articolo su rivista]
Reggiani, C.; Pellacani, G.; Reggiani Bonetti, L.; Zanelli, G.; Azzoni, P.; Chester, J.; Kaleci, S.; Ferrari, B.; Bellini, P.; Longo, C.; Bertoni, L.; Magnoni, C.
abstract

Ex-vivo fluorescence confocal microscopy (FCM) is an imaging technique that captures real time digital images of freshly excised tissue with quasi histopathological resolution. FCM reduces time to image acquisition compared to hematoxylin and eosin (H&E) frozen section analysis.

2020 - Celiac disease: histology-differential diagnosis-complications. A practical approach [Articolo su rivista]
Villanacci, V.; Vanoli, A.; Leoncini, G.; Arpa, G.; Salviato, T.; Bonetti, L. R.; Baronchelli, C.; Saragoni, L.; Parente, P.
abstract

Celiac disease is a multi-factorial chronic inflammatory intestinal disease, characterized by malabsorption resulting from mucosal injury after ingestion of wheat gluten or related rye and barley proteins. Inappropriate T-cell-mediated immune response against ingested gluten in genetically predisposed people, leads to characteristic histological lesions, as villous atrophy and intraepithelial lymphocytosis. Nevertheless, celiac disease is a comprehensive diagnosis with clinical, serological and genetic characteristics integrated with histological features. Biopsy of duodenal mucosa remains the gold standard in the diagnosis of celiac disease with the recognition of the spectrum of histological changes and classification of mucosa damage based on updated Corazza-Villanacci system. Appropriate differential diagnosis evaluation and clinical context also for the diagnosis of complications is, moreover, needed for correct histological features interpretation and clinical management.

2020 - Common bile duct lesions - how cholangioscopy helps rule out intraductal papillary neoplasms of the bile duct: A case report [Articolo su rivista]
Cocca, Silvia; Grande, Giuseppe; Reggiani Bonetti, Luca; Magistri, Paolo; Di Sandro, Stefano; Di Benedetto, Fabrizio; Conigliaro, Rita; Bertani, Helga
abstract

BACKGROUNDIntraductal papillary neoplasm of the bile duct (IPNB) is a rare variant of bile duct tumors, characterized by an exophytic growth exhibiting a papillary mass within the bile duct lumen and it can be localized anywhere along the biliary tree, with morphological variations and occasional invasion.CASE SUMMARYWe present a patient with obstructive jaundice who was diagnosed with IPNB using cholangioscopy during endoscopic retrograde cholangio-pancreatography. Using the SpyGlass DS II technology, we were able to define tumor extension and obtain targeted Spy-byte biopsies. After multidisciplinary evaluation, the patient was scheduled for surgical resection of the tumor, which was radically removed.CONCLUSIONCholangioscopy appears to be crucial for the rapid and clear diagnosis of lesions in the bile duct to achieve radical surgical resection.

2020 - Correction: PD-L1 in small bowel adenocarcinoma is associated with etiology and tumor-infiltrating lymphocytes, in addition to microsatellite instability (Modern Pathology, (2020), 33, 7, (1398-1409), 10.1038/s41379-020-0497-0) [Articolo su rivista]
Giuffrida, P.; Arpa, G.; Grillo, F.; Klersy, C.; Sampietro, G.; Ardizzone, S.; Fociani, P.; Fiocca, R.; Latella, G.; Sessa, F.; D'Errico, A.; Malvi, D.; Mescoli, C.; Rugge, M.; Nesi, G.; Ferrero, S.; Furlan, D.; Poggioli, G.; Rizzello, F.; Macciomei, M. C.; Santini, D.; Volta, U.; De Giorgio, R.; Caio, G.; Calabro, A.; Ciacci, C.; D'Armiento, M.; Rizzo, A.; Solina, G.; Martino, M.; Tonelli, F.; Villanacci, V.; Cannizzaro, R.; Canzonieri, V.; Florena, A. M.; Biancone, L.; Monteleone, G.; Caronna, R.; Ciardi, A.; Elli, L.; Caprioli, F.; Vecchi, M.; D'Inca, R.; Zingone, F.; D'Odorico, A.; Lenti, M. V.; Oreggia, B.; Bonetti, L. R.; Astegiano, M.; Biletta, E.; Cantoro, L.; Giannone, A. G.; Orlandi, A.; Papi, C.; Perfetti, V.; Quaquarini, E.; Sandri, G.; Silano, M.; Usai, P.; Barresi, V.; Ciccocioppo, R.; Luinetti, O.; Pedrazzoli, P.; Pietrabissa, A.; Viglio, A.; Paulli, M.; Corazza, G. R.; Solcia, E.; Vanoli, A.; Di Sabatino, A.
abstract

Author ‘Marco Vincenzo Lenti’ was listed in the original article as given name: ‘Marco’, family name: ‘Vincenzo Lenti’. However, this should be listed as given name: ‘Marco Vincenzo’, family name: ‘Lenti’. This has been addressed by means of this correction article as well as an update to the original article.

2020 - Digital Biopsy with Fluorescence Confocal Microscope for Effective Real-time Diagnosis of Prostate Cancer: A Prospective, Comparative Study [Articolo su rivista]
Rocco, Bernardo; Sighinolfi, Maria Chiara; Sandri, Marco; Spandri, Valentina; Cimadamore, Alessia; Volavsek, Metka; Mazzucchelli, Roberta; Lopez-Beltran, Antonio; Eissa, Ahmed; Bertoni, Laura; Azzoni, Paola; Reggiani Bonetti, Luca; Maiorana, Antonino; Puliatti, Stefano; Micali, Salvatore; Paterlini, Maurizio; Iseppi, Andrea; Rocco, Francesco; Pellacani, Giovanni; Chester, Johanna; Bianchi, Giampaolo; Montironi, Rodolfo
abstract

A microscopic analysis of tissue is the gold standard for cancer detection. Hematoxylin-eosin (HE) for the reporting of prostate biopsy (PB) is conventionally based on fixation, processing, acquisition of glass slides, and analysis with an analog microscope by a local pathologist. Digitalization and real-time remote access to images could enhance the reporting process, and form the basis of artificial intelligence and machine learning. Fluorescence confocal microscopy (FCM), a novel optical technology, enables immediate digital image acquisition in an almost HE-like resolution without requiring conventional processing.

2020 - Digital frozen section of the prostate surface during radical prostatectomy: a novel approach to evaluate surgical margins [Articolo su rivista]
Rocco, B.; Sighinolfi, M. C.; Cimadamore, A.; Reggiani Bonetti, L.; Bertoni, L.; Puliatti, S.; Eissa, A.; Spandri, V.; Azzoni, P.; Dinneen, E.; Shaw, G.; Nathan, S.; Micali, S.; Bianchi, G.; Maiorana, A.; Pellacani, G.; Montironi, R.
abstract

2020 - En-bloc resection of bladder tumors (ERBT) for pathological staging: the value of lateral margins analysis [Abstract in Rivista]
Eissa, Ahmed; Zoeir, Ahmed; Ciarlariello, Silvia; Sarchi, Luca; Sighinolfi, Maria Chiara; Ghaith, Ahmed; Puliatti, Stefano; Inzillo, Raffaele; Reggiani Bonetti, Luca; Rizzo, Mino; Rocco, Bernardo; Micali, Salvatore
abstract

In endoscopic resection of colorectal tumours, the pathological assessment of the lateral margins is a strong predictor of tumor recurrence after resection. The aims of the current study are to evaluate the value of the peritumoral margins assessment in ERBT on tumor recurrence.

2020 - Ex vivo fluorescence confocal microscopy: prostatic and periprostatic tissues atlas and evaluation of the learning curve [Articolo su rivista]
Bertoni, L.; Puliatti, S.; Reggiani Bonetti, L.; Maiorana, A.; Eissa, A.; Azzoni, P.; Bevilacqua, L.; Spandri, V.; Kaleci, S.; Zoeir, A.; Sighinolfi, M. C.; Micali, S.; Bianchi, G.; Pellacani, G.; Rocco, B.; Montironi, R.
abstract

Ex vivo fluorescence confocal microscopy (FCM) is an optical technology that provides fast H&E-like images of freshly excised tissues, and it has been mainly used for “real-time” pathological examination of dermatological malignancies. It has also shown to be a promising tool for fast pathological examination of prostatic tissues. We aim to create an atlas for FCM images of prostatic and periprostatic tissues to facilitate the interpretation of these images. Furthermore, we aimed to evaluate the learning curve of images interpretation of this new technology. Eighty fresh and unprepared biopsies obtained from radical prostatectomy specimens were evaluated using the FCM VivaScope® 2500 M-G4 (Mavig GmbH, Munich, Germany; Caliber I.D.; Rochester NY, USA) by two pathologists. Images of FCM with the corresponding H&E are illustrated to create the atlas. Furthermore, the two pathologists were asked to re-evaluate the 80 specimens after 90 days interval in order to assess the learning curve of images’ interpretation of FCM. FCM was able to differentiate between different types of prostatic and periprostatic tissues including benign prostatic glands, benign prostatic hyperplasia, high-grade intraepithelial neoplasm, and prostatic adenocarcinoma. As regards the learning curve, FCM demonstrated a short learning curve. We created an atlas that can serve as the base for urologists and pathologists for learning and interpreting FCM images of prostatic and periprostatic tissues. Furthermore, FCM images is easily interpretable; however, further studies are required to explore the potential applications of this new technology in prostate cancer diagnosis and management.

2020 - Histopathology of inflammatory bowel disease — Position statement of the Pathologists of the Italian Group for the Study of Inflammatory Bowel Disease (IG-IBD) and Italian Group of Gastrointestinal Pathologists (GIPAD-SIAPEC) [Articolo su rivista]
Villanacci, V.; Reggiani Bonetti, L.; Caprioli, F.; Saragoni, L.; Salviato, T.; Mescoli, C.; Canavese, G.; Manenti, S.; Spada, E.; Baron, L.; Leoncini, G.; Cadei, M.; Battista, S.; Armuzzi, A.
abstract

Diagnosis of the inflammatory bowel diseases ulcerative colitis (UC) and Crohn's disease (CD) relies mainly on the histopathological examination of endoscopic biopsies of the gastrointestinal tract. To facilitate the accurate diagnosis of these two conditions, this paper addresses key issues on the: (A) gastrointestinal biopsy procedure, (B) histomorphological characteristics of UC and CD, and (C) diagnosis of dysplasia. The 13 statements presented here represent the consensus of two groups of Italian pathologists (IG-IBD and GIPAD).

2020 - Laparoscopic treatment of Meckel's diverticulum perforation caused by a chicken bone. A case report [Articolo su rivista]
Manco, Gianrocco; Caramaschi, Stefania; Malagoli, Marzio; Bonetti, Luca Reggiani; Rossi, Aldo
abstract

We describe an uncommon case of a Meckel's diverticulum perforation by a chicken bone in a patient with symptoms of acute appendicitis. Meckel's diverticulum is the most common congenital abnormality of the gastrointestinal tract. The incidence is approximately 2% of the population. Most patients are asymptomatic, only 4-16% presenting complications, including bleeding, obstruction, and diverticulitis. The perforation due to a foreign body is a very infrequent complication and may have a bad prognosis in case of a delayed diagnosis. Only in a few cases, a careful evaluation of the CT scan leads to correct preoperative diagnosis. Definitive treatment is surgical intervention and should not be delayed in patients with peritonitis. Laparoscopy is a safe diagnostic and therapeutic tool to treat complicated Meckel's diverticulum. In our case, a stapled laparoscopic diverticulectomy has been performed with an excellent outcome. KEY WORDS: Laparoscopy, Meckel's diverticulum, Perforation.

2020 - Laparoscopic treatment of Meckel's diverticulum perforation caused by a chicken bone. A case report [Articolo su rivista]
Manco, G.; Caramaschi, S.; Malagoli, M.; Bonetti, L. R.; Rossi, A.
abstract

2020 - Microscopic imaging of Inflammatory Bowel Disease (IBD) and Non-IBD Colitis on digital slides: The Italian Group-IBD Pathologists experience [Articolo su rivista]
Salviato, T.; Bonetti, L. R.; Mangogna, A.; Leoncini, G.; Cadei, M.; Caprioli, F.; Armuzzi, A.; Daperno, M.; Villanacci, V.
abstract

Background: The aim of the study is to report the experience of the pathologists of the Italian Group for the Study of Inflammatory Bowel Disease (IBD) (group formed by pathologists with various experience) on the morphological assessment of digital slides pertaining to IBD and Non-IBD colitis underlining the necessity to implement this tool in daily routine and its utility to share opinions on difficult cases. Materials and Methods: Forty-eight histological slides stained with haematoxylin and eosin obtained from ileo-colorectal endoscopic biopsies were digitized using Menarini D-Sight 2.0 system, uploaded onto a website platform and shared among 40 pathologists participating in the study. Information regarding the site of biopsy was disclosed; clinical data were blinded. Each participant was committed to write a comment on microscopic features purposing diagnostic opinion. One month after the last uploaded case, a form was sent to each participant to evaluate the personal experience on digital slide sharing. Results: Sixteen pathologists out of 40 (40%) had consistently accessed to the site,9/40 (22%) commented on all slides, a diagnostic opinion was rendered in 8 slides. Most common critical issues were: A) poor internet connection resulting in ineffective evaluation of the digital slides, B) time-consuming cases raising difficult diagnostic interpretation, C) lack of clinical history. Overall, 24 participants (60%) found the forum valuable for practical training and educational purposes. Conclusions: Sharing scanned slides circulating within a dedicated forum is an effective educational tool in both IBDs and Non-IBDs colitis. Although our results demonstrated a substantial compliance of the participants, their limited participation was an objective shortcoming. Hence, further efforts are needed to encourage this potentially rewarding practice among the pathologist community.

2020 - Nodular skin lesions: correlation of reflectance confocal microscopy and optical coherence tomography features [Articolo su rivista]
Garbarino, F.; Migliorati, S.; Farnetani, F.; De Pace, B.; Ciardo, S.; Manfredini, M.; Reggiani Bonetti, L.; Kaleci, S.; Chester, J.; Pellacani, G.
abstract

Background: Nodular lesions have common clinical appearance but different prognoses. Differential diagnosis between melanoma (MM), basal cell carcinoma (BCC) and dermal naevus (DN) poses a challenge in clinical practice. Reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) are promising non-invasive imaging techniques, potentially able to decrease redundant biopsies. RCM allows in vivo visualization of skin down to the papillary dermis at almost histological resolution, while OCT, particularly dynamic OCT (D-OCT), provides images deeper within the dermis and reveals the vascular pattern. Objectives: To identify correlating features observed with RCM and OCT associated with the different nodular lesion diagnoses. Methods: We retrospectively assessed 68 nodular lesions (30 MM, 20 BCC and 18 DN) with RCM and subsequently OCT. At the end of the study, evaluations were matched with histopathological diagnosis and statistical analysis was performed. Results: In MM, 57% (17/30) evidenced both cerebriform nests at RCM and icicle-shaped structures at OCT, with higher average Breslow index. In 80% of BCCs with basaloid islands at RCM, OCT showed ovoid structures. More than half of DN (56%) showed hyporeflective nests at OCT and either dense nests or dense and sparse nests at RCM. Conclusions: The combined use of RCM and OCT offers a better understanding of the morphological architecture of nodular lesions, correlating RCM parameters with OCT and vice versa, assisting in turn with early differential diagnosis of malignant and benign nodular lesions. The correlation between icicle-shaped structures and cerebriform nests in MM and their association with Breslow index requires future research.

2020 - PD-1 blockade in deficient mismatch repair mixed adenoneuroendocrine carcinoma of the stomach: new hope for an orphan disease [Articolo su rivista]
Ricco, B.; Salati, M.; Reggiani Bonetti, L.; Dominici, M.; Luppi, G.
abstract

Introduction: Mixed adenoneuroendocrine carcinoma (MANEC) is an uncommon and aggressive tumor arising throughout the entire gastrointestinal tract. Treatment options are limited, and survival is dismal with conventional therapies. Case description: We present the case of a 66-year-old man who was diagnosed with a locally advanced MANEC of the gastroesophageal junction. He was treated with perioperative chemotherapy and total gastrectomy. After anastomotic tumor recurrence was detected, he underwent three systemic regimens, including chemoradiotherapy. The patient was then tested for mismatch repair (MMR) protein status, revealing defective expression of MLH1 and PMS2 proteins. Treatment with anti–programmed death 1 (PD-1) receptor monoclonal antibody pembrolizumab was started and radiologic response is ongoing after 11 months. Clinical benefit was evident, and no immune therapy–related side effect was detected. Conclusions: To our knowledge, this is the first report of a heavily pretreated and rapidly progressing deficient MMR gastroesophageal MANEC experiencing a durable benefit from anti-PD1 treatment.

2020 - PD-L1 in small bowel adenocarcinoma is associated with etiology and tumor-infiltrating lymphocytes, in addition to microsatellite instability [Articolo su rivista]
Giuffrida, P.; Arpa, G.; Grillo, F.; Klersy, C.; Sampietro, G.; Ardizzone, S.; Fociani, P.; Fiocca, R.; Latella, G.; Sessa, F.; D'Errico, A.; Malvi, D.; Mescoli, C.; Rugge, M.; Nesi, G.; Ferrero, S.; Furlan, D.; Poggioli, G.; Rizzello, F.; Macciomei, M. C.; Santini, D.; Volta, U.; De Giorgio, R.; Caio, G.; Calabro, A.; Ciacci, C.; D'Armiento, M.; Rizzo, A.; Solina, G.; Martino, M.; Tonelli, F.; Villanacci, V.; Cannizzaro, R.; Canzonieri, V.; Florena, A. M.; Biancone, L.; Monteleone, G.; Caronna, R.; Ciardi, A.; Elli, L.; Caprioli, F.; Vecchi, M.; D'Inca, R.; Zingone, F.; D'Odorico, A.; Lenti, M. V.; Oreggia, B.; Reggiani Bonetti, L.; Astegiano, M.; Biletta, E.; Cantoro, L.; Giannone, A. G.; Orlandi, A.; Papi, C.; Perfetti, V.; Quaquarini, E.; Sandri, G.; Silano, M.; Usai, P.; Barresi, V.; Ciccocioppo, R.; Luinetti, O.; Pedrazzoli, P.; Pietrabissa, A.; Viglio, A.; Paulli, M.; Corazza, G. R.; Solcia, E.; Vanoli, A.; Di Sabatino, A.
abstract

Small bowel adenocarcinomas (SBAs) are often associated with poor prognosis and have limited therapeutic options. Programmed cell death protein-1 (PD-1)/programmed cell death ligand 1 (PD-L1) pathway blockade is an effective treatment in many microsatellite instability-high (MSI-H) solid tumors. We aimed at investigating PD-L1 and PD-1 expression in non-hereditary, non-ampullary SBAs, associated with celiac disease (CeD), Crohn’s disease (CrD), or sporadic, recruited through the Small Bowel Cancer Italian Consortium. We assessed PD-L1 and PD-1 by immunohistochemistry in a series of 121 surgically resected SBAs, including 34 CeD-SBAs, 49 CrD-SBAs, and 38 sporadic SBAs. PD-L1 and PD-1 expression was correlated with several clinico-pathological features, such as the etiology, microsatellite instability status, and tumor-infiltrating lymphocyte (TIL) density. The prevalence of PD-L1 positivity according to combined positive score (CPS) was 26% in the whole cohort of SBAs, with significantly (p = 0.001) higher percentage (35%) in both CeD-SBAs and CrD-SBAs in comparison with sporadic SBAs (5%). CPS ≥ 1 SBAs were significantly (p = 0.013) more frequent in MSI-H cases (41%) than in non-MSI-H ones (18%); however, 15 CPS ≥ 1 microsatellite stable SBAs were also identified. CPS ≥ 1 SBAs showed higher TIL and PD-1+ immune cell density, more frequently medullary histotype, as well as a better outcome in comparison with CPS < 1 cases. This study demonstrates an increased proportion of PD-L1+ cases in both CeD-SBAs and CrD-SBAs in comparison with sporadic SBAs. In addition, the identification of a subset of PD-L1+ microsatellite stable SBAs supports the need to ascertain additional biomarkers of response to immune checkpoint inhibitors along with MSI-H.

2020 - Positive surgical margin during radical prostatectomy: overview of sampling methods for frozen sections and techniques for the secondary resection of the neurovascular bundles [Articolo su rivista]
Sighinolfi, M. C.; Eissa, A.; Spandri, V.; Puliatti, S.; Micali, S.; Reggiani Bonetti, L.; Bertoni, L.; Bianchi, G.; Rocco, B.
abstract

Objective: The aim of the paper is to provide an overview of intraoperative sampling methods for frozen section (FS) analysis and of surgical techniques for a secondary neurovascular bundle (NVB) resection, as the method of surgical margin (SM) sampling and the management of a positive SM (PSM) at the nerve-sparing (NS) area are under evaluated issues. FS analysis during radical prostatectomy (RP) can help to tailor the plane of dissection based on cancer extension and thus extend the indications for NS surgery. Evidence Acquisition: We performed a PubMed/Medical Literature Analysis and Retrieval System Online (MEDLINE), Web of Science, Cochrane Library, and Elton B. Stephens Co. (EBSCO)host search to include articles published in the last decade, evaluating FS analysis in the NS area and surgical attempts to convert a PSM to a negative status. Evidence Synthesis: Overall, 19 papers met our inclusion criteria. The ways to collect samples for FS analysis included: systematic (analysing the whole posterolateral aspect of the prostate specimen, i.e., neurovascular structure-adjacent frozen-section examination [NeuroSAFE]); magnetic resonance imaging (MRI)-guided (biopsies from MRI-suspicious areas, retrieved by the surgeon in a cognitive way); and random biopsies from the soft periprostatic tissues. Techniques to address a PSM in the NS area included: full resection of the spared NVB, from its caudal to cranial aspect, often including the rectolateral part of the Denonvilliers’ fascia; partial resection of the NVB, in cases where sampling attempts to localise a PSM; incremental approach, meaning a partial or full resection that extends until no prostate tissue is found in the soft periprostatic environment. Conclusions: There is no homogeneity in prostate sampling for FS analysis, although most recent evidence is moving toward a systematic sampling of the entire NS area. The management of a PSM is variable and can be affected by the sampling strategy (difficult localisation of the persisting tumour at the NVB). The difficult identification of the exact soft tissue location contiguous to a PSM could be considered as the critical point of FS analysis and of spared-NVB management.

2020 - Prediction of Benefit from Checkpoint Inhibitors in Mismatch Repair Deficient Metastatic Colorectal Cancer: Role of Tumor Infiltrating Lymphocytes [Articolo su rivista]
Loupakis, F.; Depetris, I.; Biason, P.; Intini, R.; Prete, A. A.; Leone, F.; Lombardi, P.; Filippi, R.; Spallanzani, A.; Cascinu, S.; Bonetti, L. R.; Maddalena, G.; Valeri, N.; Sottoriva, A.; Zapata, L.; Salmaso, R.; Munari, G.; Rugge, M.; Dei Tos, A. P.; Golovato, J.; Sanborn, J. Z.; Nguyen, A.; Schirripa, M.; Zagonel, V.; Lonardi, S.; Fassan, M.
abstract

Background: Immunotherapy with immune checkpoint inhibitors (ICIs) is highly effective in microsatellite instability–high (MSI-H) metastatic colorectal cancer (mCRC); however, specific predictive biomarkers are lacking. Patients and Methods: Data and samples from 85 patients with MSI-H mCRC treated with ICIs were gathered. Tumor infiltrating lymphocytes (TILs) and tumor mutational burden (TMB) were analyzed in an exploratory cohort of “super” responders and “clearly” refractory patients; TILs were then evaluated in the whole cohort of patients. Primary objectives were the correlation between the number of TILs and TMB and their role as biomarkers of ICI efficacy. Main endpoints included response rate (RR), progression-free survival (PFS), and overall survival (OS). Results: In the exploratory cohort, an increasing number of TILs correlated to higher TMB (Pearson's test, p =.0429). In the whole cohort, median number of TILs was 3.6 in responders compared with 1.8 in nonresponders (Mann-Whitney test, p =.0448). RR was 70.6% in patients with high number of TILs (TILs-H) compared with 42.9% in patients with low number of TILs (odds ratio = 3.20, p =.0291). Survival outcomes differed significantly in favor of TILs-H (PFS: hazard ratio [HR] = 0.42, p =.0278; OS: HR = 0.41, p =.0463). Conclusion: A significant correlation between higher TMB and increased number of TILs was shown. A significantly higher activity and better PFS and OS with ICI in MSI-H mCRC were reported in cases with high number of TILs, thus supporting further studies of TIL count as predictive biomarker of ICI efficacy. Implications for Practice: Microsatellite instability is the result of mismatch repair protein deficiency, caused by germline mutations or somatic modifications in mismatch repair genes. In metastatic colorectal cancer (mCRC), immunotherapy (with immune checkpoint inhibitors [ICIs]) demonstrated remarkable clinical benefit in microsatellite instability–high (MSI-H) patients. ICI primary resistance has been observed in approximately 25% of patients with MSI-H mCRC, underlining the need for predictive biomarkers. In this study, tumor mutational burden (TMB) and tumor infiltrating lymphocyte (TIL) analyses were performed in an exploratory cohort of patients with MSI-H mCRC treated with ICIs, demonstrating a significant correlation between higher TMB and increased number of TILs. Results also demonstrated a significant correlation between high number of TILs and clinical responses and survival benefit in a large data set of patients with MSI-H mCRC treated with ICI. TMB and TILs could represent predictive biomarkers of ICI efficacy in MSI-H mCRC and should be incorporated in future trials testing checkpoint inhibitors in colorectal cancer.

2020 - Primary laparoscopic approach to repair perforated peptic ulcer. A retrospective cohort study [Articolo su rivista]
Manco, G.; Caramaschi, S.; Rolando, G.; Malagoli, M.; Zanelli, G.; Reggiani Bonetti, L.; Rossi, A.
abstract

BACKGROUND: Perforated peptic ulcer is a serious complication of peptic ulcer disease and carries high risk for morbidity and mortality. Although the incidence of peptic ulcer disease has decreased in recent decades, the percentage of patients with perforated peptic ulcer requiring emergency surgery remains constant. The use of laparoscopic management as a first choice for the treatment of the perforation is growing but is not routine in many centers. METHODS: Clinical and surgical data on 42 patients underwent surgical treatment for perforated peptic ulcer from January 2012 to December 2016 were collected. Laparoscopic repair of the perforation with a three-port technique was made in all cases. The Boey scoring system was used to predict the prognosis. RESULTS: All patients underwent suture-closure of the ulcer, and omental patch through laparoscopy without conversion to open surgery. Duodenal leakages occurred in 3 patients (7.1%), then treated with a conservative approach and resolved on the 10th postoperative day. Two patients (4.7%) had deep space infections in the first week after surgery, therefore subdiaphragmatic and pelvic abscess were drained by ultrasound guidance. Four patients (9.5%) died up to 30-day postsurgery due to progression of multisystem organ failure in absence of leakages or infections. All these patients were American Society of Anesthesiologists Classification >III and Boey Score 3. CONCLUSIONS: Our data show that a primary laparoscopic approach in patients with peptic ulcer perforation is associated with postoperative advantages and acceptable rates of morbidity and mortality, essentially related to high Boey Score. Therefore, we suggest that the surgical repair of PPU could be always started laparoscopically.

2020 - Real-time assessment of surgical margins during radical prostatectomy: a novel approach that uses fluorescence confocal microscopy for the evaluation of peri-prostatic soft tissue [Articolo su rivista]
Rocco, B.; Sighinolfi, M. C.; Bertoni, L.; Spandri, V.; Puliatti, S.; Eissa, A.; Reggiani Bonetti, L.; Azzoni, P.; Sandri, M.; De Carne, C.; Turri, F.; Cimadamore, A.; Montironi, R.; Maiorana, A.; Micali, S.; Bianchi, G.; Pellacani, G.
abstract

2020 - Rectal-cancer radiotherapy damages the perineal muscle floor [Articolo su rivista]
Reggiani Bonetti, L.; Manenti, A.; Gallo, G.; Domati, F.
abstract

2020 - Scleral ossicles: Angiogenic scaffolds, a novel biomaterial for regenerative medicine applications [Articolo su rivista]
Checchi, M.; Bertacchini, J.; Cavani, F.; Magaro, M. S.; Reggiani Bonetti, L.; Pugliese, G. R.; Tamma, R.; Ribatti, D.; Maurel, D. B.; Palumbo, C.
abstract

Given the current prolonged life expectancy, various pathologies affect increasingly the aging subjects. Regarding the musculoskeletal apparatus, bone fragility induces more susceptibility to fractures, often not accompanied by good ability of self-repairing, in particular when critical-size defects (CSD) occur. Currently orthopedic surgery makes use of allografting and autografting which, however, have limitations due to the scarce amount of tissue that can be taken from the donor, the possibility of disease transmission and donor site morbidity. The need to develop new solutions has pushed the field of tissue engineering (TE) research to study new scaffolds to be functionalized in order to obtain constructs capable of promoting tissue regeneration and achieve stable bone recovery over time. This investigation focuses on the most important aspect related to bone tissue regeneration: the angiogenic properties of the scaffold to be used. As an innovative solution, scleral ossicles (SOs), previously characterized as natural, biocompatible and spontaneously decellularized scaffolds used for bone repair, were tested for angiogenic potential and biocompatibility. To reach this purpose, in ovo Chorioallantoic Membrane Assay (CAM) was firstly used to test the angiogenic potential; secondly, in vivo subcutaneous implantation of SOs (in a rat model) was performed in order to assess the biocompatibility and the inflammatory response. Finally, thanks to the analysis of mass spectrometry (LCMSQE), the putative proteins responsible for the SO angiogenic properties were identified. Thus, a novel natural biomaterial is proposed, which is (i) able to induce an angiogenic response in vivo by subcutaneous implantation in a non-immunodeficient animal model, (ii) which does not induce any inflammatory response, and (iii) is useful for regenerative medicine application for the healing of bone CSD.

2020 - Two-Stage Turnbull-Cutait Pull-Through Coloanal Anastomosis for Low Rectal Cancers [Articolo su rivista]
Manenti, A.; Reggiani Bonetti, L.; Manco, G.
abstract

2020 - “Real-time” Assessment of Surgical Margins During Radical Prostatectomy: State-of-the-Art [Articolo su rivista]
Eissa, A.; Zoeir, A.; Sighinolfi, M. C.; Puliatti, S.; Bevilacqua, L.; Del Prete, C.; Bertoni, L.; Azzoni, P.; Reggiani Bonetti, L.; Micali, S.; Bianchi, G.; Rocco, B.
abstract

Histopathologic examination of the pathologic specimens using hematoxylin & eosin stains represents the backbone of the modern pathology. It is time-consuming; thus, “real-time” assessment of prostatic and periprostatic tissue has gained special interest in the diagnosis and management of prostate cancer. The current study focuses on the review of the different available techniques for “real-time” evaluation of surgical margins during radical prostatectomy (RP). We performed a comprehensive search of the Medline database to identify all the articles discussing “real-time” or intraoperative assessment of surgical margins during RP. Several filters were applied to the search to include only English articles performed on human subjects and published between January 2000 and March 2019. The search revealed several options for pathologic assessment of surgical margins including intraoperative frozen sections, confocal laser endomicroscopy, optical spectroscopy, photodynamic diagnosis, optical coherence tomography, multiphoton microscopy, structured illumination microscopy, 3D augmented reality, and ex vivo fluorescence confocal microscope. Frozen section represents the gold standard technique for real-time pathologic examinations of surgical margins during RP; however, several other options showed promising results in the initial clinical trials, and considering the rapid development in the field of molecular and cellular imaging, some of these options may serve as an alternative to frozen section.

2019 - Class 1, 2, and 3 BRAF-mutated metastatic colorectal cancer: A detailed clinical, pathologic, and molecular characterization [Articolo su rivista]
Schirripa, M.; Biason, P.; Lonardi, S.; Pella, N.; Simona Pino, M.; Urbano, F.; Antoniotti, C.; Cremolini, C.; Corallo, S.; Pietrantonio, F.; Gelsomino, F.; Cascinu, S.; Orlandi, A.; Munari, G.; Malapelle, U.; Saggio, S.; Fontanini, G.; Rugge, M.; Mescoli, C.; Lazzi, S.; Bonetti, L. R.; Lanza, G.; Dei Tos, A. P.; De Maglio, G.; Martini, M.; Bergamo, F.; Zagonel, V.; Loupakis, F.; Fassan, M.
abstract

Purpose: BRAF mutations are grouped in activating RASindependent signaling as monomers (class 1-V600E) or as dimers (class 2-codons 597/601), and RAS-dependent with impaired kinase activity (class 3-codons 594/596). Although clinical, pathologic, and molecular features of V600EBRAFmutated metastatic colorectal cancer (mCRC) are well known, limited data are available from the two other classes. Experimental Design: Data from 117 patients with BRAF (92 class 1, 12 class 2, and 13 class 3)-mutated mCRC were collected. A total of 540 BRAF wt mCRCs were included as control. IHC profiling was performed to determine the consensus molecular subtypes (CMS), cytokeratin 7/20 profiles, tumor-infiltrating lymphocyte infiltration, and BM1/BM2 categorization. Overall survival (OS) and progression-free survival were evaluated by Kaplan-Meier and log-rank test. Results: Class 3 BRAF-mutated mCRC was more frequently left sided (P = 0.0028), pN0 (P = 0.0159), and without peritoneal metastases (P = 0.0176) compared with class 1, whereas class 2 cases were similar to class 1. Hazard ratio for OS, as compared with BRAF wt, was 2.38 [95% confidence interval (CI), 1.61-3.54] for class 1, 1.90 (95% CI, 0.85-4.26) for class 2, and 0.93 (95% CI, 0.51-1.69) for class 3 (P < 0.0001). Class 2 and 3 tumors were all assigned to CMS2-3. A higher median CD3/CD8-positive lymphocyte infiltration was observed in BRAF-mutated class 2 (P = 0.033) compared with class 3 cases. Conclusions: For the first time, different clinical and pathologic features and outcome data were reported according to the three BRAF mutation classes in mCRC. Specific targeted treatment strategies should be identified in the near future for such patients.

2019 - Development and validation of confocal endomicroscopy diagnostic criteria for low-grade dysplasia in barrett’s esophagus [Articolo su rivista]
Di Pietro, M.; Bertani, H.; O'Donovan, M.; Santos, P.; Alastal, H.; Phillips, R.; Ortiz-Fernandez-sordo, J.; Iacucci, M.; Modolell, I.; Bonetti, L. R.; Ragunath, K.; Wernisch, L.
abstract

OBJECTIVES: Low-grade dysplasia (LGD) in Barrett's esophagus (BE) is generally inconspicuous on conventional and magnified endoscopy. Probe-based confocal laser endomicroscopy (pCLE) provides insight into gastro-intestinal mucosa at cellular resolution. We aimed to identify endomicroscopic features and develop pCLE diagnostic criteria for BE-related LGD. METHODS: This was a retrospective study on pCLE videos generated in 2 prospective studies. In phase I, 2 investigators assessed 30 videos to identify LGD endomicroscopic features, which were then validated in an independent video set (n = 25). Criteria with average accuracy >80% and interobserver agreement κ > 0.4 were taken forward. In phase II, 6 endoscopists evaluated the criteria in an independent video set (n = 57). The area under receiver operating characteristic curve was constructed to find the best cutoff. Sensitivity, specificity, interobserver, and intraobserver agreements were calculated. RESULTS: In phase I, 6 out of 8 criteria achieved the agreement and accuracy thresholds (i) dark nonround glands, (ii) irregular gland shape, (iii) lack of goblet cells, (iv) sharp cutoff of darkness, (v) variable cell size, and (vi) cellular stratification. The best cutoff for LGD diagnosis was 3 out of 6 positive criteria. In phase II, the diagnostic criteria had a sensitivity and specificity for LGD of 81.9% and 74.6%, respectively, with an area under receiver operating characteristic of 0.888. The interobserver agreement was substantial (κ = 0.654), and the mean intraobserver agreement was moderate (κ = 0.590). CONCLUSIONS: We have generated and validated pCLE criteria for LGD in BE. Using these criteria, pCLE diagnosis of LGD is reproducible and has a substantial interobserver agreement.

2019 - Development of a Desmocollin-3 Active Mouse Model Recapitulating Human Atypical Pemphigus [Articolo su rivista]
Lotti, Roberta; Atene, Claudio Giacinto; Marconi, Alessandra; Di Rocco, Giulia; Reggiani Bonetti, L; Zanocco Marani, Tommaso; Pincelli, Carlo
abstract

Pemphigus vulgaris (PV) is a life-threatening mucocutaneous autoimmune blistering disease. It is often associated with autoantibodies to the desmosomal adhesion proteins Desmoglein 3 (DSG3) and Desmoglein 1 (DSG1). Recently, auto-antigens, such as desmocollins and others have been described in PV and in atypical pemphigus forms such as Pemphigus Herpetiformis (PH), Pemphigus Vegetans (PVeg), and Paraneoplastic Pemphigus (PP). Desmocollins belong to a cadherin subfamily that provides structure to the desmosomes and play an important role in cell-to-cell adhesion. In order to verify the pathogenic activity of anti-Desmocollin 3 (DSC3) antibodies, we developed an active disease model of pemphigus expressing anti-DSC3 autoantibodies or antiDSC3 and anti-DSG3 antibodies. This approach included the adoptive transfer of DSC3 and/or DSG3 lymphocytes to Rag2(-/-) immunodeficient mice that express DSC3 and DSG3. Our results show that the presence of anti-DSC3 auto-antibodies is sufficient to determine the appearance of a pathological phenotype relatable to pemphigus, but with features not completely super-imposable to those observed in the DSG3 active model, suggesting that the DSC3 active model might mimic the atypical pemphigus. Moreover, the presence of both anti-DSC3 and anti-DSG3 antibodies determines a more severe phenotype and a slower response to prednisolone. In conclusion, we have developed an adult DSC3 pemphigus mouse model that differs from the DSG3 model and supports the concept that antigens other than desmogleins may be responsible for different phenotypes in human pemphigus.

2019 - Ex-vivo Fluorescence Confocal Microscopy: The First Application For Real-Time Pathologic Examination of Prostatic Tissue [Articolo su rivista]
Puliatti, Stefano; Bertoni, Laura; Pirola, Giacomo Maria; Azzoni, Paola; Bevilacqua, Luigi; Eissa, Ahmed; Elsherbiny, Ahmed; Sighinolfi, Maria Chiara; Chester, Johanna; Rocco, Bernardo; Micali, Salvatore; Bagni, Ilaria; Reggiani Bonetti, Luca; Maiorana, Antonino; Malvehy, Josep; Longo, Caterina; Montironi, Rodolfo; Bianchi, Giampaolo; Pellacani, Giovanni
abstract

Prostate cancer "real time" intra-operative pathological examination currently utilizes frozen section, despite the many inherent limitations of this procedure. Ex vivo Fluorescence Confocal Microscopy (FCM) is a novel technology, which provides fast microscopic fluorescence and reflectance tissue imaging. FCM has been proven beneficial in colorectal, breast, thyroid and skin lesions. However, it was never applied in urology. The aim of our study is to report the first application of FCM in the urologic field assessing its diagnostic accuracy for non-neoplastic and cancerous prostate tissue (prostatic adenocarcinoma) compared to the gold standard histopathological diagnoses.

2019 - External validation of a novel side-specific, multiparametric magnetic resonance imaging-based nomogram for the prediction of extracapsular extension of prostate cancer: preliminary outcomes on a series diagnosed with multiparametric magnetic resonance imaging-targeted plus systematic saturation biopsy [Articolo su rivista]
Sighinolfi, Maria Chiara; Sandri, Marco; Torricelli, Pietro; Ligabue, Guido; Fiocchi, Federica; Scialpi, Michele; Eissa, Ahmed; Reggiani Bonetti, Luca; Puliatti, Stefano; Bianchi, Giampaolo; Rocco, Bernardo
abstract

no abstract

2019 - Lynch syndrome with exclusive skin involvement: time to consider a molecular definition? [Articolo su rivista]
Vaisfeld, A.; Calicchia, M.; Pomponi, M. G.; Lucci-Cordisco, E.; Reggiani Bonetti, L.; Genuardi, M.
abstract

Muir–Torre syndrome (MTS) is clinically characterized by the occurrence of skin, usually sebaceous, and visceral tumors in the same individual. The most common underlying mechanism is a constitutional defect of the mismatch repair (MMR) genes that cause Lynch syndrome (LS). Herewithin we report on a 76 years-old male patient heterozygous for a pathogenic MSH2 missense substitution who presented with a striking cutaneous phenotype in the absence of typical LS visceral tumors. The patient developed 20 skin tumors, including sebaceous adenomas/carcinomas and keratoacanthomas. Two skin tumors showed immunohistochemical loss of MSH2 and MSH6 expression. There was no apparent family history of neoplasia. Based on the variable involvement of the skin and internal organs, we suggest that the definition of tumor associations that are often observed as variants of inherited tumor syndromes, such as MTS, should be guided by the underlying molecular bases. In addition, the presence of multiple sebaceous tumors, especially if showing MMR deficiency, appears to be a very strong indicator of a constitutional MMR gene defect. The reasons underlying the high phenotypic variability of cutaneous phenotypes associated with constitutional MMR defects are yet to be determined.

2019 - Massive juvenile polyposis of the stomach in a family with SMAD4 gene mutation [Articolo su rivista]
de Leon, M. P.; Pedroni, M.; Viel, A.; Luppi, C.; Conigliaro, R.; Domati, F.; Rossi, G.; Bonetti, L. R.
abstract

Relatively little is known on the genotype-phenotype correlations between SMAD4 gene mutations, juvenile polyposis of the intestine and Hereditary Hemorrhagic Teleangectasia. We describe a family in which the proband (a 46-year old woman) had massive polyposis of the stomach—leading to surgery—with high-grade dysplasia at histology. Molecular analysis was carried out using Next Generation sequencing techniques with Miseq Illumina Platforms and a minimal coverage of 40 reads. In the proband, the analysis showed the presence of a truncating mutation in the SMAD4 gene (c.1213dupC, a variant previously associated with juvenile polyposis and Hereditary Hemorrhagic Teleangectasia). The same mutation was detected in two other members of the family (father and brother of the proband), who showed massive polypoid involvement of the stomach at gastroscopy. By taking the family history, subtle evidence of Hereditary Teleangectasia was found (nasal bleeding and arterovenous malformations) in the three gene carriers. Colonoscopy showed polyp occurrence in all three affected members with SMAD4 mutation, with prevalence of adenomatous lesions in one (father), of hamartomas in the brother, and of a mix of histological types in the proband. The main features of the family can be summarized as follows: (A) In hereditary juvenile polyposis, lesions of different histology can be detected at colonoscopy; (B) In the gene carriers of SMAD4 mutations, lesions of the stomach require careful surveillance and, when necessary, surgical interventions; (C) Signs and symptoms of Hereditary Hemorrhagic Teleangectasia should be suspected (and searched) in individuals with SMAD4 constitutional mutations.

2019 - Melanoma types by in vivo reflectance confocal microscopy correlated with protein and molecular genetic alterations: A pilot study [Articolo su rivista]
Beretti, F.; Bertoni, L.; Farnetani, F.; Pellegrini, C.; Gorelli, G.; Cesinaro, A. M.; Reggiani Bonetti, L.; Di Nardo, L.; Kaleci, S.; Chester, J.; Longo, C.; Massi, D.; Fargnoli, M. C.; Pellacani, G.
abstract

Cutaneous melanoma (CM) is one of the most prevalent skin cancers, which lacks both a prognostic marker and a specific and lasting treatment, due to the complexity of the disease and heterogeneity of patients. Reflectance confocal microscopy (RCM) in vivo analysis is a versatile approach offering immediate morphological information, enabling the identification of four primary cutaneous RCM CM types. Whether RCM CM types are associated with a specific protein and molecular genetic profiles at the tissue level remains unclear. The current pilot study was designed to identify potential correlations between RCM CM types and specific biological characteristics, combining immunohistochemistry (IHC) and molecular analyses. Eighty primary CMs evaluated at patient bedside with RCM (type 1 [19, 24%], type 2 [12, 15%], type 3 [7, 9%] and type 4 [42, 52%]) were retrospectively evaluated by IHC stains (CD271, CD20, CD31, cyclin D1), fluorescence in situ hybridization FISH for MYC gain and CDKN2A loss and molecular analysis for somatic mutations (BRAF, NRAS and KIT). RCM CM types correlated with markers of stemness property, density of intra-tumoral lymphocytic B infiltrate and cyclin D1 expression, while no significant association was found with blood vessel density nor molecular findings. RCM CM types show a different marker profile expression, suggestive of a progression and an increase in aggressiveness, according to RCM morphologies.

2019 - Morphological classification of melanoma metastasis with reflectance confocal microscopy [Articolo su rivista]
Farnetani, F; Manfredini, M; Longhitano, S; Chester, J; Shaniko, K; Cinotti, E; Mazzoni, L; Venturini, M; Manganoni, A; Longo, C; Reggiani-Bonetti, L; Giannetti, L; Rubegni, P; Calzavara-Pinton, P; Stanganelli, I; Perrot, J L; Pellacani, G
abstract

Cutaneous malignant melanoma metastases differential diagnosis is challenging, as clinical and dermoscopic features can simulate primary melanoma or other benign or malignant skin neoplasms, and in-vivo reflectance confocal microscopy could assist. Our aim was to identify specific reflectance confocal microscopy features for cutaneous malignant melanoma metastases, and epidermal and dermal involvement.

2019 - Morphology and molecular features of rare colorectal carcinoma histotypes [Articolo su rivista]
Remo, A.; Fassan, M.; Vanoli, A.; Bonetti, L. R.; Barresi, V.; Tatangelo, F.; Gafa, R.; Giordano, G.; Pancione, M.; Grillo, F.; Mastracci, L.
abstract

Several histopathological variants of colorectal carcinoma can be distinguished, some associated with specific molecular profiles. However, in routine practice, ninety/ninety-five percent of all large bowel tumors are diagnosed as conventional adenocarcinoma, even though they are a heterogeneous group including rare histotypes, which are often under-recognized. Indeed, colorectal cancer exhibits differences in incidence, location of tumor, pathogenesis, molecular pathways and outcome depending on histotype. The aim is therefore to review the morphological and molecular features of these rare variants of intestinal carcinomas which may hold the key to differences in prognosis and treatment.

2019 - NUT midline carcinoma: Current concepts and future perspectives of a novel tumour entity [Articolo su rivista]
Salati, M.; Baldessari, C.; Bonetti, L. R.; Messina, C.; Merz, V.; Cerbelli, B.; Botticelli, A.
abstract

NMC is a recently recognized cancer type hallmarked by chromosomal translocation involving the NUT gene, a catastrophic event leading to fusion oncoprotein responsible for malignant transformation and tumor progression. The aggressiveness of disease together with a poor response to conventional treatment make NMC one of the most lethal cancer. Moreover, although until recently NMC has been poorly understood and largely neglected, the number of reported cases is steadily rising. Recently, in addition to its pathogenetic and diagnostic role, NUT-fusion oncoprotein has been shown to be amenable to targeted inhibition using BET inhibitors. Future clinical trials are warranted with the aim of investigate the incorporation of targeted agent into multimodal therapeutic strategy. Since new promising NUT-targeting drugs are emerging that may affect the clinical course, the correct and prompt recognition of NMC is key to improve patients' outcome.

2019 - Poorly differentiated clusters (PDC) in colorectal cancer: Does their localization in tumor matter? [Articolo su rivista]
Bertoni, Laura; Barresi, Valeria; REGGIANI BONETTI, Luca; Caramaschi, Stefania; Mangogna, Alessandro; Lionti, Simona; Azzoni, Paola; Carnevale, Gianluca; Pisciotta, Alessandra; Salviato, Tiziana
abstract

2019 - The A.L.A.N. score identifies prognostic classes in advanced biliary cancer patients receiving first-line chemotherapy [Articolo su rivista]
Salati, M.; Caputo, F.; Cunningham, D.; Marcheselli, L.; Spallanzani, A.; Rimini, M.; Gelsomino, F.; Reggiani Bonetti, L.; Andrikou, K.; Rovinelli, F.; Smyth, E.; Baratelli, C.; Kouvelakis, K.; Kalaitzaki, R.; Gillbanks, A.; Michalarea, V.; Cascinu, S.; Braconi, C.
abstract

Background: Chemotherapy is the mainstay treatment for advanced biliary cancer (ABC). Best supportive care and clinical trials are currently alternative options. The identification of a prognostic score that can be widely applied to daily practice has the potential to better inform clinical management of ABC patients. Methods: A cohort of 123 ABC patients undergoing first-line chemotherapy was used as an exploratory cohort to define the prognostic value of laboratory tests routinely performed in clinical practice. Kaplan–Meier analysis was used to investigate the association between the variables and overall survival (OS). Those variables that were statistically significant at the multivariate analysis were combined in a multiplex score. Performance of the novel prognostic score was confirmed in a validation cohort of 60 ABC patients. Results: Baseline actual neutrophil count, lymphocytes-monocytes ratio, neutrophil-lymphocytes ratio and albumin (A.L.A.N.) correlated with OS at the multivariate analysis in the exploratory cohort. When combined in the multiplex, A.L.A.N. score was able to identify three classes of ABC patients with significantly different OS (high-risk: median OS, 5 months; intermediate-risk: median OS, 12 months and low-risk: median OS, 22 months; p:<0.001). The score performed well in the different subtypes of ABC and was independent of stage, performance status and chemotherapy regimen. The performance of the A.L.A.N. score was confirmed in a validation cohort of cholangiocarcinoma patients (high-risk: median OS, 4.3 months; intermediate-risk: median OS 9.3 months, low-risk: median OS 13 months; p:0.005). Conclusions: The A.L.A.N score can be derived by variables routinely recorded in clinical practice and can provide prognostic assessment of ABC patients considered for first-line treatment.

2019 - The histopathologic report of surgically resected colorectal liver metastases: What is clinically relevant? [Articolo su rivista]
Barresi, V.; Fioravanzo, A.; Pecori, S.; Tomezzoli, A.; Reggiani Bonetti, L.
abstract

Colorectal carcinoma (CRC) is one of the most common malignancies and a major cause of cancer-related death worldwide. The liver is the most frequent site of metastatic spread, so that about half of the patients with CRC have or develop liver metastases (LM) during the clinical course of the disease. Colorectal LM can potentially be cured by surgery, but most patients still experience disease progression and recurrence after the surgical treatment. Prediction of a patient's post-surgical clinical course is mainly based on clinical parameters or the histopathological features of the primary tumor, while little attention is given to the pathological characteristics of the LM. In this paper, we review the prognostic relevance of the gross and microscopic pathological features observed in surgically resected LM and propose which information should be included in the histopathological report to guide surgeons and oncologists for the subsequent therapeutic management.

2018 - Clinical impact and prognostic role of KRAS/BRAF/PIK3CA mutations in stage I colorectal cancer [Articolo su rivista]
Bonetti, L. R.; Barresi, V.; Maiorana, A.; Manfredini, S.; Caprera, C.; Bettelli, S.
abstract

Stage I colorectal carcinoma has excellent prognosis, with 5-year survival rate up to 95%. The occurrence of lymphovascular invasion, tumor budding, high number of PDC, or lymph node micrometastases is associated with tumor progression. The aim of this study was to evaluate the mutational status of 62 stage I colorectal carcinomas (CRC) (taken from 37 patients surviving more than five years since the initial diagnosis and from 25 patients who died of disease) and to correlate it with histopathological features and the clinical outcome. Mutations of KRAS, NRAS, BRAF, and PIK3CA genes were analyzed through Myriapod Colon Status Kit, using the high-throughput genotyping platform Sequenom MassARRAY System. Mutations in those genes were found in 31 cases (50%) and mainly in those with poor prognosis. The most frequent mutations occurred at codons 12 and 13 of the KRAS gene (40% of cases). We found concomitant PIK3CA mutations in 5 cases (8%). The presence of PIK3CA mutations was mainly observed in tumors with poor prognosis and with unfavorable histopathological prognostic features. High PDC grade (P = 0 0112), the presence of tumor budding (P = 0 0334), LVI (P < 0 0001), KRAS mutations (P = 0 0228), PIK3CA mutations (P = 0 0214), multiple genetic mutations in KRAS and PIK3CA genes (P = 0 039), and nodal micrometastases (P < 0 0001) were significant prognostic variables for CSS. The presence of LVI was the only independent and statistically significant prognostic variable for CSS in our cohort of pTNM stage I CRCs. The analysis of KRAS/PIK3CA mutational status may be used to identify patients with stage I CRC at high risk of bad outcome and who may need additional treatments, including biological therapies.

2018 - Energy drinks and obesity: Preliminary results from a preclinical study [Articolo su rivista]
Mattioli, Anna Vittoria; Manenti, Antonio; Bonetti, Luca Reggiani; Farinetti, Alberto
abstract

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2018 - Ex vivo fluorescence confocal microscopy for intraoperative, real-time diagnosis of cutaneous inflammatory diseases: A preliminary study [Articolo su rivista]
Bertoni, L; Azzoni, P; Reggiani, C; Pisciotta, A; Carnevale, G; Chester, J; Kaleci, S; Reggiani Bonetti, L; Cesinaro, Am; Longo, C; Pellacani, G.
abstract

Ex vivo fluorescence confocal microscopy (FCM) is an innovative imaging tool that can be used intraoperatively to obtain real-time images of untreated excised tissue with almost histologic resolution. As inflammatory diseases often share overlapping clinical features, histopathology evaluation is required for dubious cases, delaying definitive diagnoses, and therefore therapy. This study identifies key-features at ex vivo FCM for differential diagnoses of cutaneous inflammatory diseases, in particular, psoriasis, eczema, lichen planus and discoid lupus erythematosus. Retrospective ex vivo FCM and histological evaluations with relevant diagnoses were correlated with prospectively reported histopathologic diagnoses, to evaluate agreement and the level of expertise required for correct diagnoses. We demonstrated that ex vivo FCM enabled the distinction of the main inflammatory features in most cases, providing a substantial concordance to histopathologic diagnoses. Moreover, ex vivo FCM and histological evaluations reached a substantial agreement with histopathologic diagnoses both for all raters and for each operator. After a yet to be defined learning curve, these preliminary results suggest that dermatologists may be able to satisfactorily interpret ex vivo FCM images for correct real-time diagnoses. Despite some limitations mainly related to the equipment of FCM with a single objective lens, our study suggests that ex vivo FCM seems a promising tool in assisting diagnoses of cutaneous inflammatory lesions, with a level of accuracy quite close to that offered by histopathology. This is the first study to investigate ex vivo FCM application in cutaneous inflammatory lesions, and to evaluate the diagnostic capability of this technology.

2018 - Histology of the mesorectal lymphatics explains aspects of rectal cancer [Articolo su rivista]
Reggiani-Bonetti, L.; Barresi, V.; Manenti, A.; Domati, F.; Farinetti, A.
abstract

2018 - Histopathological variables in liver metastases of patients with stage IV colorectal cancer: potential prognostic relevance of poorly differentiated clusters [Articolo su rivista]
Lionti, S.; Reggiani Bonetti, L.; Bettelli, S.; Spallanzani, A.; Gelsomino, F.; Barresi, V.
abstract

The prognosis of patients with colorectal liver metastases (LMs) is mostly established on clinical variables or on the anatomic extent of colorectal cancer (CRC). Histopathological factors of LMs which may actually reflect the biological aggressiveness of the tumor are not routinely considered to define the risk of worse clinical outcome in those patients. The number of poorly differentiated clusters (PDCs) of neoplastic cells in primary CRC is associated with metastatic risk and bad prognosis, but PDC presence in LMs has been barely analyzed thus far. We assessed PDC presence in the histological slides of surgically resected and synchronous LMs in 63 patients with CRC who had been not submitted to any neoadjuvant treatments. Then, we analyzed its association with patients’ cancer-specific survival (CSS) or progression-free survival. The presence of PDCs (P =.016) and PDC localization at tumor edge of LMs (P =.0004) were significantly associated with shorter CSS. PDC presence at the periphery of LMs and positive resection margin were independent prognostic variables for CSS. PDC localization at the tumor edge of LMs was a significant (P =.0079) and independent prognosticator of shorter progression-free survival. Our data suggest that PDC presence and peripheral localization in LMs may be relevant to predict outcome and useful for clinical decision making in patients with colorectal synchronous LMs.

2018 - Optimal processing of ESD specimens to avoid pathological artifacts [Articolo su rivista]
Reggiani Bonetti, L.; Manta, R.; Manno, M.; Conigliaro, R.; Missale, G.; Bassotti, G.; Villanacci, V.
abstract

Background: En bloc endoscopic submucosal dissection (ESD) has been recently introduced as a treatment for precancerous/neoplastic gastrointestinal conditions. The aim of the present study was histological assessment of en bloc ESD specimens. Methods: Fifty-three ESD specimens were positioned over a cellulose acetate support (40 specimens; 12 from the upper gastrointestinal tract and 28 from the lower gastrointestinal tract) or pinned with nails on polystyrene or cork (13 specimens; 7 from the upper gastrointestinal tract and 6 from the lower gastrointestinal tract). We cut consecutive 2 mm-thick sections stained with hematoxylin and eosin. From the first and the last sections, we obtained a second slide, after a 180° rotation and re-embedding. The quality of ESD samples was scored as inadequate, suboptimal and adequate, based on the amount of crushing, shearing and stretching artifacts that were scored from 0 (absent) to 2 (diffuse or maximum). From the sum of these we obtained a global artifact score (GAS). Results: Removed lesions were: adenocarcinoma (5 cases), neuroendocrine tumor (NET) G1 (1 case), premalignant conditions, including adenomatous polyps (41 cases) and hyperplastic lesions (6 cases). A positive deep surgical margin was found in 8/53 cases (15%): high- and low-grade dysplastic glands were detected in 5 cases, low-grade adenocarcinoma in 2, and NET cells in 1. Dysplastic glands were detected in the lateral surgical margins of 12 ESD specimens (23%). Among the ESD specimens positioned on the cellulose acetate support, apart from the modifications due to electrocoagulation, 2 (5%) showed shearing modifications. In the group of ESD specimens fixed with nails, 5 (38%) showed shearing, 10 (77%) crushing artifacts, 11 (85%) stretching and 11 (85%) multiple holes caused by the nails. On the basis of these data all histological specimens from ESD on cellulose acetate were adequate (GAS 0–1).However, in the group of ESD fixed with nails, 1 was adequate (GAS 0), 11 suboptimal (GAS 2–5) and 1 inadequate (GAS 6). Conclusions: Specific devices including cellulose support and adequate sampling blocks can be helpful to perform accurate histological assessment of ESD specimens after en bloc ESD for precancerous/neoplastic gastrointestinal lesions, with complete analysis of the status of the margins and the entirely en bloc evaluation of the lesion.

2018 - Poorly differentiated clusters in colorectal liver metastases: Prognostic significance in synchronous and metachronous metastases [Articolo su rivista]
Barresi, V.; Lionti, S.; Bonetti, L. R.
abstract

2018 - Radiation therapy damages external anal sphincter and perineal muscle floor [Articolo su rivista]
Domati, F.; Bonetti, L. R.; Barresi, V.; Zizzo, M.; Manenti, A.
abstract

2018 - Reinterpreting dermoscopic pigment network with reflectance confocal microscopy for identification of melanoma-specific features [Articolo su rivista]
De Pace, B.; Farnetani, F.; Losi, A.; Ciardo, S.; De Carvalho, N.; Cesinaro, A. M.; Reggiani Bonetti, L.; Chester, J.; Kaleci, S.; Del Duca, E.; Nisticò, S. P.; Longo, C.; Pellacani, G.
abstract

Background: Pigment network is an important dermoscopic feature for melanocytic lesions, but alterations in grid line thickness are also observed in melanomas. Objective: To investigate features of thick, thin and mixed pigment networks at dermoscopy and their respective features at reflectance confocal microscopy (RCM) for differential diagnosis, correlated with histology. Methods: All melanocytic lesions with histological diagnosis, evaluated between January 2010 and May 2014, were enrolled and classified according to dermoscopy evaluation of the pigment networks: thin, thick and mixed. Results: Thin network in melanoma was characterized by a honeycombed pattern (P < 0.001), dendritic cells (P < 0.001), atypical ringed pattern (P = 0.035) and structureless area (P = 0.012), whereas round cells (P < 0.001), dendritic cells (P < 0.001) and atypical meshwork pattern (<0.001) characterized thick network in melanoma. Mixed network type in melanoma shared honeycombed (P = 0.049) and typical ringed patterns (P = 0.045) in the thin area and round cells (P < 0.001) and atypical meshwork pattern (P < 0.001) in the thick area. Thin network in nevi was characterized by cobblestone (P < 0.001) and typical ringed patterns (P = 0.035), whereas thick network in nevi showed a typical meshwork pattern (P < 0.001). Mixed nevi shared the same features and patterns, but more frequently with inflammatory infiltrate (P = 0.047). Conclusion: Differential diagnosis between melanocytic lesions (nevi or melanoma) in thin, thick and mixed pigment networks observed at dermoscopy can be assisted by RCM to improve diagnostic accuracy.

2018 - Risk of colorectal polyps and of malignancies in asymptomatic carriers of mutations in the main DNA mismatch repair genes [Articolo su rivista]
Ponz de Leon, Maurizio; Pedroni, Monica; Pezzi, Annalisa; Sulce, Blerta; Roncucci, Luca; Domati, Federica; Rossi, Giuseppina; Reggiani Bonetti, Luca
abstract

Objective: Mutation carriers (Mut+) in DNA mismatch repair genes are predisposed to cancer of various organs and to adenomatous polyps; however, they may remain asymptomatic and cancer or polyp-free for several years. We purposed to analyse the clinical follow-up of individuals carrying constitutional mutations in the MLH1, MSH2 or MSH6 genes who were unaffected by benign polyps or malignant tumours at diagnosis. Material and Methods: Mut + subjects (n.81) were members of Lynch syndromes in whom mutations were detected between 1993 and 2015; all were asymptomatic at diagnosis. They were informed of the cancer risk and surveillance was suggested. As controls, 113 nongene carriers (Mutâ) in the same Lynch families were identified. Results: About one-fourth of the mutation carriers developed polyps, mostly adenomas; polyps were less (12%, p <.05) in Mut â subjects, and hyperplastic lesions were the prevalent histology. More polyps were detected in MLH1 vs. MSH2 mutation carriers. In Mut+, 21 malignant tumours developed in 14 carriers vs. 4 tumours in 3 patients among Mutâ (p <.001). Tumours were mostly of the Lynch spectrum; however, three glioblastomas were developed, together with neoplasms of various organs (duodenum, thyroid, skin, lung and cervix). Mean age of tumour occurrence was 43.0 years in Mut + vs. 53.0 among Mutâ. Conclusions: Cancer developed more often in Mut+, with no consistent difference between MLH1 and MSH2 carriers. More polyps (mostly adenomas) were detected in MLH1 carriers. The majority (13 of 21) of malignant tumours occurred in organs for which there is no recommended surveillance, and were lethal in three patients.

2017 - A Dose-finding and Biomarker Evaluation Phase Ib Study of Everolimus in Association With 5-Fluorouracil and Pelvic Radiotherapy as Neoadjuvant Treatment of Locally Advanced Rectal Cancer (E-LARC Study) [Articolo su rivista]
Gelsomino, Fabio; Bertolini, Federica; Luppi, Gabriele; Spallanzani, Andrea; Pettorelli, Elisa; Reggiani Bonetti, Luca; Meduri, Bruno; Manco, Gianrocco; Conte, Pierfranco; Cascinu, Stefano
abstract

Micro-Abstract Neoadjuvant chemoradiation represents the mainstay of the treatment of locally advanced rectal cancer (LARC). However, despite substantial improvements in previous years, patients still develop relapses and eventually die of metastatic disease. In the present phase Ib dose-escalation trial, everolimus added to standard chemoradiation did not seem to worsen toxicity. The activity of this combination warrants further evaluation in larger clinical trials. Background During the past 20 years, considerable improvement has occurred in the treatment of patients with locally advanced rectal cancer (LARC). With the introduction of multimodal treatment, refinements in preclinical staging and improvements in surgical skills, local relapse is no longer the major problem for patients with LARC. However, many patients die of metastatic disease. The present phase Ib study aimed to establish the maximum tolerated dose of everolimus combined with 5-fluorouracil and radiotherapy in patients with LARC. Patients and Methods Patients were sequentially assigned to 4 cohorts with an increasing dose of everolimus, starting from 14 days before 5-fluorouracil and radiotherapy and continuing throughout concomitant treatment. The secondary endpoints were the Dworak tumor regression grade, pathologic complete response rate, neoadjuvant rectal score, biomarker assessment (phosphorylated mTOR [mammalian target of rapamycin] protein and phosphorylated-p70S6K protein). Results At the time of this report, 12 patients had been treated, and no dose-limiting toxicity was recorded. The most frequently reported acute toxicities were rectal tenesmus, skin rash, diarrhea, and dysuria. All 12 patients underwent curative R0 resection. Two patients had Dworak tumor regression grade 4 (pathologic complete response). No everolimus-related postoperative complications were observed. No relationship was found between biomarker expression and the clinicopathologic outcomes. Conclusion Although the addition of everolimus did not appear to worsen the toxicity of chemoradiation in patients with LARC, evaluation of its activity deserves further investigation in larger clinical trials.

2017 - Analysis of KRAS, NRAS, PIK3CA, and BRAF mutational profile in poorly differentiated clusters of KRAS-mutated colon cancer [Articolo su rivista]
Reggiani Bonetti, Luca; Barresi, Valeria; Bettelli, Stefania; Caprera, Cecilia; Manfredini, Samantha; Maiorana, Antonio
abstract

Recently, a grading system based on the counting of poorly differentiated clusters (PDCs) of neoplastic cells was shown to be a strong predictor of nodal metastases and negative prognosis in colon cancer (CC). In this study, we assessed and compared the mutational status of KRAS, NRAS, and PIK3CA in PDCs and corresponding main tumor tissue of 25 CCs with KRAS mutations. For each tumor, PDC and main tumor tissue were distinctly analyzed by using laser microdissection and mass spectrometry. In 3 CCs, the main tumor tissue had also PIK3CA mutations (C420R: 1; E545K: 1; H1047R: 1), and in 1, it showed NRAS mutation (codon 12). In 20 cases, PDCs had the same biomolecular profile as the main tumor, but in 5, they had different biomolecular profiles. In detail, PDCs had KRAS wild type in 2 cases and additional PIK3CA mutations (E542K: 1; H1047Y: 1; E545Q: 1) in 3. All 3 cases with additional PIK3CA mutations in PDCs had nodal metastases, high pathological TNM stage, and lymphatic invasion. In 1 of 3 cases, additional PIK3CA mutation detected in PDC, but not in the main tumor, was also found in the corresponding nodal metastases. Our findings show for the first time that heterogeneous biomolecular profile previously observed in CC may depend on different histologic aspects of the lesion. Because PDCs may represent the tumor cells with the highest potential to metastatize, their molecular status may be relevant for the prediction of response to targeted therapies.

2017 - Attenuated polyposis of the large bowel: a morphologic and molecular approach [Articolo su rivista]
PONZ DE LEON, Maurizio; Pedroni, Monica; Roncucci, Luca; Domati, Federica; Rossi, Giuseppina; Magnani, Giulia; Pezzi, Annalisa; Fante, Rossella; Bonetti, Luca Reggiani
abstract

Attenuated polyposis could be defined as a variant of familial adenomatous polyposis (FAP) in which synchronous polyps of the large bowel range between 10 and 99. We analysed all cases of attenuated polyposis observed over the last 30 years with the objectives: (A) to classify the disease according to different type and proportion of polyps; (B) To ascertain the contribution of APC and MutYH genes; (C) to discover features which could arise the suspicion of mutations; (D) To obtain indications for management and follow-up. 84 individuals in 82 families were studied. Polyps were classified into four groups as adenoma, hyperplastic, other serrated lesions or others; APC and MutYH mutations were assessed. Mean age at diagnosis was 54 ± 14 years in men and 48 ± 13 in women (P = 0.005). Polyps were more numerous in women (37 ± 26 vs 29 ± 22). Sixty % of patients underwent bowel resection, mainly for cancer; the remaining were managed through endoscopy. A total of 2586 polyps were detected at diagnostic endoscopy: 2026 (80 %) were removed and analysed. Adenomas were diagnosed in 1445 (70 %), hyperplastic polyps in 541 (26 %), other serrated lesions in 61 (2.9 %). Adenomas and hyperplastic lesions were detected in the majority of patients. In 68 patients (81 %) in whom studies were executed, APC mutations were found in 8 and MutYH mutations in 10. Genetic variants were more frequent in women (12 vs 6, P = 0.039). Taking into consideration the prevalent (>50 %) histology and presence of mutations, patients could be subdivided into four groups: (1) APC mutated polyposis (AFAP), when adenomas were >50 % and APC mutations detected (no. 8, 10 %); (2) MutYH mutated polyposis (MAP), adenomas >50 % and biallelic MutYH mutations (no. 10, 12 %); (1) attenuated polyposis without detectable mutations, prevalence of adenomas, 48 cases (57 %); (1) hyperplastic-serrated polyposis, with prevalence (>50 %) of hyperplastic/other serrated lesions and no constitutional mutation (no. 18, 21 %). Aggregation of tumors, cancer in probands, distribution of polyps and other clinical characteristics showed no difference among the four groups. In conclusions, AFAP and MAP, the polyposis labeled by constitutional mutations, represented about 25 % of all attenuated polyposis. Mutation-associated cases showed an earlier age of onset of polyps and were more frequent in the female sex.

2017 - Cytohistological and immunohistochemical characteristics of spindle-shaped mesenchymal neoplasms occurring in the gastrointestinal tract [Articolo su rivista]
Ieni, Antonio; Barresi, Valeria; Reggiani Bonetti, Luca; Branca, Giovanni; Caruso, Rosario Alberto; Tuccari, Giovanni
abstract

The purpose of the present review is to analyze the cytohistological and immunohistochemical characteristics of spindle-shaped mesenchymal gastrointestinal neoplams (MGNs), a group of unusual neoplastic conditions with different biological behavior. These tumors exhibit clinical pictures strictly related to the site of origin and dimensions, even if they appear generally with an intramural localization. This latter point may suggest an useful application of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA), mainly followed by the cell-block procedure (CBP) in the differential diagnostic approach. First of all, we discuss the most common entity of MGNs represented by gastrointestinal stromal tumors (GISTs), analyzing the morphologic characteristics and stressing the strength of immunohistochemical algorithm for diagnostic purposes. Successively, we have reported the less common group of spindle-shaped MGNs comprehensive of those arising elsewhere the soft tissues, such as leiomyomas, leiomyosarcomas, schwannomas, inflammatory myofibroblastic tumor and intra-abdominal desmoid fibromatosis. Finally, very uncommon spindle-shaped MGNs, like clear cell, follicular dendritic cell, undifferentiated pleomorphic and radiation-induced sarcomas as well as spindle cell dedifferentiated liposarcomas, have been briefly mentioned.

2017 - Do pathological variables have prognostic significance in rectal adenocarcinoma treated with neoadjuvant chemoradiotherapy and surgery? [Articolo su rivista]
Bonetti, Luca Reggiani; Lionti, Simona; Domati, Federica; Barresi, Valeria
abstract

AIM To clarify which factors may influence pathological tumor response and affect clinical outcomes in patients with locally advanced rectal carcinoma treated with neo-adjuvant chemoradiotherapy and surgery. METHODS Tumor regression grade (TRG) according to the Dworak system and yTNM stage were assessed and correlated with pre-treatment clinico-pathological variables in 215 clinically locally advanced (cTNM stage â¡U and â¡V) rectal carcinomas. Prognostic value of all pathological and clinical factors on disease free survival (DFS) and cancer specific survival (CSS) was analyzed by Kaplan Meier and Cox-regression analyses. RESULTS cN+ status, mucinous histotype or poor differentiation in the pre-treatment biopsy were significantly associated with lower pathological response (low Dworak grade and TNM remaining unchanged/upstaging). Cases showing acellular mucin pools in surgical specimens all had unremarkable clinical courses with no deaths or recurrences during follow-up. Dworak grade had prognostic significance for DFS and CSS. However, compared to the 5-tiered system, a simplified twotiered grading system, in which grades 0, 1 and 2 were grouped as absent/partial regression and grades 3 and 4 were grouped as total/subtotal regression, was more reproducible and prognostically informative. The twotiered Dworak system, yN stage, craniocaudal extension of the tumor and radial margin status were significant independent prognostic variables. CONCLUSION Our data suggest that caution should be applied in using a conservative approach in rectal carcinomas with cN+ status, extensive/lower involvement of the rectum and mucinous histotype or poor differentiation. Although Dworak TRG is prognostically significant, a simplified two-tiered system could be preferable. Finally, cases with acellular mucin pools should be carefully evaluated to definitely exclude residual mucinous carcinoma.

2017 - Does stem cell therapy induce myocardial neoangiogenesis? Histological evaluation in an ischemia/reperfusion animal model [Articolo su rivista]
Pennella, Sonia; Bonetti, L. R.; Migaldi, Mario; Manenti, Antonio; Lonardi, Roberto; Giuliani, Enrico; Barbieri, Alberto; Farinetti, Alberto; Mattioli, Anna Vittoria
abstract

Background: In an experimental model in the rabbit, a myocardial ischemia-reperfusion injury was obtained. Subsequently, the effects of homologous bone marrow stem cell (BMSC) administration were studied. Methods: In 21 New Zealand adult rabbits, ischemia/reperfusion damage was induced by temporary occlusion of the anterior descending coronary artery. Homologous BMSCs were isolated, cultured and re-suspended for injection at the level of the ischemic zone. We evaluated the proangiogenetic effect of intramyocardial injections of BMSC at the peri-infarcted area. Histological evaluations were made after 20 days from the surgical procedure. Results: In rabbits treated with intramyocardial BMSC administration, we demonstrated histologically capillary neoangiogenesis, without signs of tissue immunological reaction or of generation of new myocardial cells. On the contrary, only minimal neovascular supply was detected in rabbits treated with intravenous administration of BMSC. Only typical signs of ischemic myocardium injury were observed in the control group. Conclusion: These observations suggest that the effect of direct BMSC administration in ischemic myocardium could promote a capillary neoangiogenesis, which helps to prevent ischemic myocardial damage.

2017 - Does topical hemostatic agent (FlosealÂ®) have a long-term adverse effect on erectile function recovery after nerve-sparing robot-assisted radical prostatectomy? [Articolo su rivista]
Martorana, Eugenio; Rocco, Bernardo; Kaleci, Shaniko; Pirola, Giacomo Maria; Bevilacqua, Luigi; Bonetti, Luca Reggiani; Puliatti, Stefano; Micali, Salvatore; Bianchi, Giampaolo
abstract

Objectives: To investigate the long-term effects of FlosealÂ® on erectile function recovery (EFR) after nerve-sparing robot-assisted radical prostatectomy (RALP). Methods: We prospectively collected results of the self-administered International Index Erectile Function Questionnaire 1â5 and 15 (IIEF 1â5 and 15) of 532 consecutive patients who underwent RALP for prostate cancer in our institution between October 2007 and December 2015. Patients were divided into two groups according to FlosealÂ® application after prostatectomy. They were enrolled according to the following criteria: (a) bilateral nerve-sparing procedure; (b) preoperative IIEFÂ â¥Â 17; adherence to our erectile rehabilitation protocol; (c) 1-year follow-up. Outcomes were measured as mean IIEF score, EFR (IIEFÂ 25). Results: A total of 120 patients were enrolled. Group A included 40 consecutive patients who received traditional hemostasis, and Group B included 80 consecutive patients in which FlosealÂ® was additionally used. No differences were observed in terms of preoperative mean IIEF score (pÂ =Â 0.65). Group B patients showed a trend toward a higher mean IIEF score 3Â months after surgery (pÂ =Â 0.06) but no differences in terms of EFR (pÂ =Â 1.000). Long-term results (6, 9, 12Â months after surgery) showed a significantly and progressively higher mean IIEF score (pÂ =Â 0.04, 0.003, 0.003) and EFR (pÂ =Â 0.043, 0.027, 0.004) in Group A patients. Comparison between the groups in terms of severe, moderate, mild and no ED becomes significant at 9 and 12Â months (pÂ =Â 0.002, 0.006). Conclusion: The results of our study suggest that local use of FlosealÂ® worsens the long-term erectile function recovery in patients selected for nerve-sparing RALP.

2017 - Heavily calcified gastrointestinal stromal tumors: Pathophysiology and implications of a rare clinicopathologic entity [Articolo su rivista]
Salati, M.; Orsi, G.; Bonetti, L. R.; Di Benedetto, F.; Longo, G.; Cascinu, S.
abstract

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract, and are characterized by a broad spectrum of clinical, histological and molecular features at presentation. Although focal and scattered calcifications are not uncommon within the primary tumor mass, heavy calcification within a GIST is rarely described in the literature and the clinical-biological meaning of this feature remains unclear. Cases with such an atypical presentation are challenging and may be associated with diagnostic pitfalls. Herein, we report a gastric GIST with the unusual presentation of prominent calcifications that was identified incidentally on imaging during a post-trauma diagnostic work-up. The patient underwent laparoscopic surgery with a radical resection of the mass, which was subsequently characterized by histological analysis as spindle-shaped tumor cells, positive for CD117/c-KIT, CD34 and DOG1, and with calcified areas. Given the intermediate risk of recurrence, no adjuvant therapy was recommended and the patient underwent regular follow-up for 22 mo, with no evidence of relapse. Our case can be considered of interest because of the rarity of clinical presentation and the uniquely large size of the GIST at diagnosis (longest diameter exceeding 9 cm). In closing, we discuss the pathophysiology and clinical implications of calcifications in GISTs by reviewing the most up-to-date relevant literature.

2017 - Histological grading based on poorly differentiated clusters is predictive of tumour response and clinical outcome in rectal carcinoma treated with neoadjuvant chemoradiotherapy [Articolo su rivista]
Reggiani Bonetti, Luca; Lionti, Simona; Domati, Federica; Pagliani, Giuliana; Mattioli, Elisabetta; Barresi, Valeria
abstract

Aims: The clinical outcome of patients with locally advanced rectal cancer who undergo neoadjuvant chemoradiotherapy (CRT) is influenced by the tumour response to treatment, which is reflected by tumour regression grade and post-treatment (y) TNM stage. Little is known about the prognostic value of pretreatment histopathological features of the tumour that may be useful to discriminate potential non-responders and to design tailored therapeutic strategies. In this study, we aimed to investigate the prognostic role of poorly differentiated clusters (PDCs) of neoplastic cells in pretreatment biopsies of patients with rectal cancer treated with neoadjuvant CRT. Methods and results: Grading based on PDC counting was retrospectively applied to 204 pretreatment endoscopic biopsies of rectal carcinomas from patients treated with neoadjuvant CRT and surgery. Interobserver agreement in the assessment of PDC grade was good. High PDC grade was significantly associated with high yT stage (PÂ =Â 0.044), yM+ status (PÂ =Â 0.0004), and unchanged TNM stage or TNM upstaging (PÂ =Â 0.032). In addition, high PDC grade was a significant and independent prognostic factor for cancer-specific survival. Conclusions: PDC grade may be assessed in preoperative biopsies of rectal cancer with good reproducibility. High PDC grade in a pretreatment tumour is significantly associated with a poor response to therapy. Hence, we suggest that PDC grading might be used as a significant predictive and prognostic factor in patients with locally advanced rectal cancer who are treated with neoadjuvant CRT, and to identify high-risk patients who need surgery and adjuvant chemotherapy.

2017 - Interpathologist concordance in the histological diagnosis of focal prostatic atrophy lesions, acute and chronic prostatitis, PIN, and prostate cancer [Articolo su rivista]
Giunchi, Francesca; Jordahl, Kristina; Bollito, Enrico; Colecchia, Maurizio; Patriarca, Carlo; D’Errico, Antonietta; Vasuri, Francesco; Malvi, Deborah; Fornari, Alessandro; Bonetti, Luca Reggiani; Corti, Barbara; Papotti, Mauro; DeGiuli, Paolo; Loda, Massimo; Montironi, Rodolfo; Fiorentino, Michelangelo; Rider, Jennifer R.
abstract

Epidemiological and biological evidence indicates a causal relationship between the presence of proliferative atrophic lesions and the development of prostatic intraepithelial neoplasia (PIN) and prostate cancer. The presence of inflammatory and atrophic lesions of the prostate is widely underestimated and they are not generally mentioned in pathology reports. We performed a histopathological concordance study among eight genitourinary specialists and seven generalist pathologists, using 116 histological slides of prostate lesions, including proliferative atrophic lesions, PIN, and cancer. The overall agreement between all possible pairs of reviewers was 80% for prostate cancer, 67% for PIN, and 49% for proliferative atrophic lesions. When using as gold standard the assessment of a single genitourinary pathologist, the mean agreement percentage increased to 97% for prostate cancer, 92% for PIN, and 72% for proliferative atrophic lesions.

2017 - Lesion volume predicts prostate cancer risk and aggressiveness: validation of its value alone and matched with prostate imaging reporting and data system score [Articolo su rivista]
Martorana, Eugenio; Pirola, Giacomo Maria; Scialpi, Michele; Micali, Salvatore; Iseppi, Andrea; Bonetti, Luca Reggiani; Kaleci, Shaniko; Torricelli, Pietro; Bianchi, Giampaolo
abstract

Objective: To demonstrate the association between magnetic resonance imaging (MRI) estimated lesion volume (LV), prostate cancer detection and tumour clinical significance, evaluating this variable alone and matched with Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) score. Patients and methods: We retrospectively analysed 157 consecutive patients, with at least one prior negative systematic prostatic biopsy, who underwent transperineal prostate MRI/ultrasonography fusion-targeted biopsy between January 2014 and February 2016. Suspicious lesions were delineated using a ‘region of interest’ and the system calculated prostate volume and LV. Patients were divided in groups considering LV (≤0.5, 0.5–1, ≥1 mL) and PI-RADS score (1–5). We considered clinically significant prostate cancer as all cancers with a Gleason score of ≥3 + 4 as suggested by PI-RADS v2. A direct comparison between MRI estimated LV (MRI LV) and histological tumour volume (HTV) was done in 23 patients who underwent radical prostatectomy during the study period. Differences between MRI LV and HTV were assessed using the paired sample t-test. MRI LV and HTV concordance was verified using a Bland–Altman plot. The chi-squared test and logistic and ordinal regression models were used to evaluate difference in frequencies. Results: The MRI LV and PI-RADS score were associated both with prostate cancer detection (both P < 0.001) and with significant prostate cancer detection (P < 0.001 and P = 0.008, respectively). When the two variables were matched, increasing LV increased the risk within each PI-RADS group. Prostate cancer detection was 1.4-times higher for LVs of 0.5–1 mL and 1.8-times higher for LVs of ≥1 mL; significant prostate cancer detection was 2.6-times for LVs of 0.5–1 mL and 4-times for LVs of ≥1 mL. There was a positive correlation between MRI LV and HTV (r = 0.9876, P < 0.001). Finally, Bland–Altman analysis showed that MRI LV was underestimated by 4.2% compared to HTV. Study limitations include its monocentric and retrospective design and the limited cohort. Conclusions: This study demonstrates that PI-RADS score and the MRI LV, independently and in combination, are associated with prostate cancer detection and with tumour clinical significance.

2017 - Mesocolic micro-skip metastasis [Articolo su rivista]
REGGIANI BONETTI, Luca; Manenti, Antonio; Domati, Federica; Farinetti, Alberto
abstract

The presence of micro skip metastases point out the interest of complete mesocolon excision.

2017 - Poorly Differentiated Clusters: Clinical Impact in Colorectal Cancer [Articolo su rivista]
Barresi, Valeria; Reggiani Bonetti, Luca; Ieni, Antonio; Caruso, Rosario Alberto; Tuccari, Giovanni
abstract

Colorectal cancer (CRC) is one of the most common malignancies worldwide and it still represents a major cause of cancer-related death. Postsurgical Tumor Node Metastasis (pTNM) stage is the main prognostic factor in CRC and it currently drives therapeutic protocols after surgery. However, CRC outcome is not always predictable on the basis of pTNM stage. Hence, there is the need to find additional prognostic factors that may allow to predict the clinical course of CRC regardless of pTNM stage, so that patients may receive the most appropriate treatment for their tumor. In recent years, the prognostic value of a novel grading system based on the counting of poorly differentiated clusters (PDCs) of neoplastic cells in tumor tissue was documented in CRC. In this article, we review the clinical relevance of grading based on PDC counting in CRC. In view of its high prognostic value and reproducibility, PDC grading could be introduced in routine histopathological report of CRC and used for therapeutic decision making. Although there is initial evidence that PDC may be correlated with biomolecular profile of CRC, whether their presence is associated with response to targeted therapies is still unknown and deserves further investigation.

2017 - Prognostic significance of CDX2 immunoexpression in poorly differentiated clusters of colorectal carcinoma [Articolo su rivista]
Reggiani Bonetti, Luca; Lionti, Simona; Vitarelli, Enrica; Barresi, Valeria
abstract

CDX2 is a transcription factor that acts as a tumor suppressor in colorectal cancer (CRC). Its loss triggers metastatic process and tumor progression; however, its prognostic role in patients with CRC is still controversial. Poorly differentiated clusters (PDCs) are aggregates of neoplastic cells which likely have high metastatic potential in CRC. In this study, we analyzed and compared CDX2 expression in PDC (CDX2-PDC) and corresponding main tumor (CDX2 main tumor) in 42 CRCs showing at least 10 PDC (PDC G3). Five of 42 CRCs (12%) were classified as CDX2 main tumor negative (4/5 were also PDC-CDX2 negative); all had tumor recurrence and died of CRC. Twenty nine of 42 cases were CDX2-PDC negative. Among CRC CDX2 main tumor positive, 15 had recurrences and 13 died from CRC; 13 and 11 of them, respectively, were CDX2-PDC negative. By assigning one point to CDX2 main tumor or CDX2-PDC positivity, we assessed CDX2-staining score for each case. Twelve cases had CDX2-staining score 2 (CDX2 positive in main tumor and PDC); 26 had score 1 (CDX2 positive in main tumor or PDC), and 4 had CDX2 score 0 (CDX2 negative in main tumor and PDC). In our patients, CDX2-staining score had higher prognostic value compared to CDX2 main tumor or CDX2-PDC alone. In addition, it represented a significant and independent prognostic variable for disease-free survival (DFS) and cancer-specific survival (CSS). Our findings suggest that, although loss of CDX2 in the main tumor identifies high-risk patients with high specificity, CDX2-PDC should also be considered in CDX2 main tumor positive cases to predict prognosis.

2017 - Small bowel carcinomas in celiac or Crohn's disease: Distinctive histophenotypic, molecular and histogenetic patterns [Articolo su rivista]
Vanoli, A.; Di Sabatino, A.; Martino, M.; Klersy, C.; Grillo, F.; Mescoli, C.; Nesi, G.; Volta, U.; Fornino, D.; Luinetti, O.; Fociani, P.; Villanacci, V.; D'armiento, F. P.; Cannizzaro, R.; Latella, G.; Ciacci, C.; Biancone, L.; Paulli, M.; Sessa, F.; Rugge, M.; Fiocca, R.; Corazza, G. R.; Solcia, E.; Ardizzone, S.; Astegiano, M.; Caio, G.; Calabro, A.; Canzonieri, V.; Cerutti, R.; Ciardi, A.; Coppola, L.; D'inca, R.; Elli, L.; Ferrero, S.; Florena, A. M.; Furlan, D.; Giannone, A. G.; Giuffrida, P.; Macciomei, M. C.; Maccioni, A.; Monteleone, G.; Migliora, P.; Orlandi, A.; Papi, C.; Perfetti, V.; Bonetti, L. R.; Rizzo, A.; Salemme, M.; Sandri, G.; Sampietro, G.; Santeusanio, G.; Santini, D.; Silano, M.; Solina, G.; Tonelli, F.; Trapani, D.; Usai, P.
abstract

Non-familial small bowel carcinomas are relatively rare and have a poor prognosis. Two small bowel carcinoma subsets may arise in distinct immune-inflammatory diseases (celiac disease and Crohn's disease) and have been recently suggested to differ in prognosis, celiac disease-associated carcinoma cases showing a better outcome, possibly due to their higher DNA microsatellite instability and tumor-infiltrating T lymphocytes. In this study, we investigated the histological structure (glandular vs diffuse/poorly cohesive, mixed or solid), cell phenotype (intestinal vs gastric/pancreatobiliary duct type) and Wnt signaling activation (β-catenin and/or SOX-9 nuclear expression) in a series of 26 celiac disease-associated small bowel carcinoma, 25 Crohn's disease-associated small bowel carcinoma and 25 sporadic small bowel carcinoma cases, searching for new prognostic parameters. In addition, non-tumor mucosa of celiac and Crohn's disease patients was investigated for epithelial precursor changes (hyperplastic, metaplastic or dysplastic) to help clarify carcinoma histogenesis. When compared with non-glandular structure and non-intestinal phenotype, both glandular structure and intestinal phenotype were associated with a more favorable outcome at univariable or stage- and microsatellite instability/tumor-infiltrating lymphocyte-inclusive multivariable analysis. The prognostic power of histological structure was independent of the clinical groups while the non-intestinal phenotype, associated with poor outcome, was dominant among Crohn's disease-associated carcinoma. Both nuclear β-catenin and SOX-9 were preferably expressed among celiac disease-associated carcinomas; however, they were devoid, per se, of prognostic value. We obtained findings supporting an origin of celiac disease-associated carcinoma in SOX-9-positive immature hyperplastic crypts, partly through flat β-catenin-positive dysplasia, and of Crohn's disease-associated carcinoma in a metaplastic (gastric and/or pancreatobiliary-type) mucosa, often through dysplastic polypoid growths of metaplastic phenotype. In conclusion, despite their common origin in a chronically inflamed mucosa, celiac disease-associated and Crohn's disease-associated small bowel carcinomas differ substantially in histological structure, phenotype, microsatellite instability/tumor-infiltrating lymphocyte status, Wnt pathway activation, mucosal precursor lesions and prognosis.

2017 - Small bowel carcinomas in coeliac or Crohn's disease: Clinico-pathological, molecular, and prognostic features. A study from the small bowel cancer Italian consortium [Articolo su rivista]
Vanoli, Alessandro; Di Sabatino, Antonio; Furlan, Daniela; Klersy, Catherine; Grillo, Federica; Fiocca, Roberto; Mescoli, Claudia; Rugge, Massimo; Nesi, Gabriella; Fociani, Paolo; Sampietro, Gianluca; Ardizzone, Sandro; Luinetti, Ombretta; Calabrò, Antonio; Tonelli, Francesco; Volta, Umberto; Santini, Donatella; Caio, Giacomo; Giuffrida, Paolo; Elli, Luca; Ferrero, Stefano; Latella, Giovanni; Ciardi, Antonio; Caronna, Roberto; Solina, Gaspare; Rizzo, Aroldo; Ciacci, Carolina; D'Armiento, Francesco P.; Salemme, Marianna; Villanacci, Vincenzo; Cannizzaro, Renato; Canzonieri, Vincenzo; Bonetti, Luca Reggiani; Biancone, Livia; Monteleone, Giovanni; Orlandi, Augusto; Santeusanio, Giuseppe; Macciomei, Maria C.; D'Incà, Renata; Perfetti, Vittorio; Sandri, Giancarlo; Silano, Marco; Florena, Ada M.; Giannone, Antonino G.; Papi, Claudio; Coppola, Luigi; Usai, Paolo; Maccioni, Antonio; Astegiano, Marco; Migliora, Paola; Manca, Rachele; Martino, Michele; Trapani, Davide; Cerutti, Roberta; Alberizzi, Paola; Riboni, Roberta; Sessa, Fausto; Paulli, Marco; Solcia, Enrico; Corazza, Gino R.
abstract

Background and Aims: An increased risk of small bowel carcinoma [SBC] has been reported in coeliac disease [CD] and Crohn's disease [CrD]. We explored clinico-pathological, molecular, and prognostic features of CD-associated SBC [CD-SBC] and CrD-associated SBC [CrD-SBC] in comparison with sporadic SBC [spo-SBC]. Methods: A total of 76 patients undergoing surgical resection for non-familial SBC [26 CD-SBC, 25 CrD-SBC, 25 spo-SBC] were retrospectively enrolled to investigate patients' survival and histological and molecular features including microsatellite instability [MSI] and KRAS/NRAS, BRAF, PIK3CA, TP53, HER2 gene alterations. Results: CD-SBC showed a significantly better sex-, age-, and stage-adjusted overall and cancerspecific survival than CrD-SBC, whereas no significant difference was found between spo-SBC and either CD-SBC or CrD-SBC. CD-SBC exhibited a significantly higher rate of MSI and median tumour-infiltrating lymphocytes [TIL] than CrD-SBC and spo-SBC. Among the whole SBC series, both MSI-which was the result of MLH1 promoter methylation in all but one cases-and high TIL density were associated with improved survival at univariable and stage-inclusive multivariable analysis. However, only TILs retained prognostic power when clinical subgroups were added to the multivariable model. KRAS mutation and HER2 amplification were detected in 30% and 7% of cases, respectively, without prognostic implications. Conclusions: In comparison with CrD-SBC, CD-SBC patients harbour MSI and high TILs more frequently and show better outcome. This seems mainly due to their higher TIL density, which at multivariable analysis showed an independent prognostic value. MSI/TIL status, KRAS mutations and HER2 amplification might help in stratifying patients for targeted anti-cancer therapy.

2017 - Solid pseudopapillary tumor: a new tumor entity in the testis? [Articolo su rivista]
Mengoli, Maria Cecilia; Bonetti, Luca Reggiani; Intersimone, Donatella; Fedeli, Franco; Rossi, Giulio
abstract

We greatly appreciated the case by Michal et al [1] reporting on a pancreatic analogue solid pseudopapillary tumor (SPT) of the paratesticular location. Analogously, we recently dealt with 2 cases arising in the right testis of a 60-year-old man and in the left testis of a 56-year-old man. The patients have an uneventful medical history, and the testicular nodules (1.5 and 3.2 cm, respectively) were incidentally disclosed during an ultrasound echosonography performed for hydrocele. Routine laboratory test results and serum tumor markers were unremarkable.

2017 - The miR-196b miRNA inhibits the GATA6 intestinal transcription factor and is upregulated in colon cancer patients [Articolo su rivista]
Fantini, Sebastian; Salsi, Valentina; Reggiani, Luca; Maiorana, Antonino; Zappavigna, Vincenzo
abstract

Objective: To explore the possible misexpression of the microRNA miR-196b in colorectal cancer (CRC) and its role in controlling the expression of GATA6, a putative target gene crucial to intestinal cell homeostasis and tumorigenesis. Design: The expression of miR-196b was analysed by qRT-PCR in surgical resection samples from a cohort of sporadic colon cancer patients. Manipulations of miR-196b expression were performed to demonstrate its inhibition of GATA6 protein levels. Results: We found that miR-196b is significantly upregulated in pre-treatment surgical resection samples from a cohort of sporadic colon cancer patients. The upregulation of miR-196b correlates with less severe clinicopathological characteristics, such as early tumor stage and absence of lymph node metastases. We show that in CRC cells, miR-196b targets the mRNA of GATA6, a transcription factor involved in the homeostasis and differentiation of intestinal epithelial cells, and a positive regulator of the Wnt/β-catenin pathway. We moreover found that the increase of miR-196b correlates with a reduced GATA6 protein expression in colon cancer patients. Conclusion: Our results establish miR-196b as a post-transcriptional inhibitor of GATA6 in CRC cells, implicating miR-196b function in gene regulatory pathways crucial to intestinal cell homeostasis and tumorigenesis. Our results furthermore suggest a role of miR-196b expression in CRC, as an antagonist of GATA6 function in tumor cells, thus providing the basis for a potential targeting strategy for the treatment of CRC.

2017 - Total mesocolon examination in colon cancer [Articolo su rivista]
Reggiani Bonetti, Luca; Domati, Federica; Manenti, Antonio; Farinetti, Alberto
abstract

We read with interest the article by Bertelsen et al1 concerning the mesocolic lymph node excision. We think that, in the case of colon cancers, a complete histological examination of the entire mesocolon allows for the discovery of important features of its secondary pathology. Following this policy in 400 consecutive cases of T2 and T3 colon cancers, we observed in 6 cases (1.5%) microskip secondary lesions in lymphatic structures at the mesenteric root, in proximity with the lumboaortic stations. This agrees with the particular arrangement of the lymphatic network of the mesocolon, where the capillaries are predominant and through numerous interconnections open different and unexpected ways to lymph drainage, under the action of different and variable external forces.2 In 2 cases, the secondary microskip metastases were placed in newly formed lymphatic capillaries, suggesting that the neolymphangiogenesis, induced by the tumor, favors implantation of metastases.3 Interestingly, in 2 patients the microskip metastases appeared to be located inside mesocolic lymphatic aggregates, where lymphocytes CD20+ B prevail on the CD4+ T-helper and CD8+ T-cytotoxic cells. These structures, already present in the normal mesocolon and in the greater omentum, lack proper capsule and follicular organization.4,5 In patients with colon cancer, they can increase in number and show a progressive evolution toward proper lymph nodes; we interpret this as a sign of immunological reaction against the tumor.

2017 - Toward optimal processing of endoscopic submucosal dissection specimens [Articolo su rivista]
Villanacci, Vincenzo; Bassotti, Gabrio; Bonetti, Luca Reggiani; Trecca, Antonello; Cadei, Moris
abstract

Abstract non available.

2016 - Clinical significance of pelvic lymph node status in prostate cancer: review of 1690 cases [Articolo su rivista]
Maccio, Livia; Barresi, Valeria; Domati, Federica; Martorana, Eugenio; Cesinaro, Anna Maria; Migaldi, Mario; Iachetta, Francesco; Ieni, Antonio; Bonetti, Luca Reggiani
abstract

To assess whether any relationship exists between the number of histologically examined lymph nodes and the detection of metastases in pelvic lymph node dissection (PLND) specimens taken from patients with radical prostatectomy (RP) for prostatic adenocarcinoma. 1690 cases of RP with PNLD were included in the study; 54Â % of the patients were submitted to extended PLND (ePLND). KaplanâMeier curves confirm the negative prognostic significance of nodal metastases on the overall patientsâ survival (PÂ 

2016 - Determination of Human Chorionic Gonadotropin in Postmortem Samples in Ectopic Pregnancies [Articolo su rivista]
Palmiere, Cristian; del Mar Lesta, Maria; Fanton, Laurent; Ventura, Francesco; Bonsignore, Alessandro; Reggiani Bonetti, Luca
abstract

Increased human chorionic gonadotropin levels (HCG) can be detected in femoral blood, bile, and vitreous humor collected during autopsy of pregnant women using a standard kit designed for living patients. In the study herein, the concentrations of HCG were measured in postmortem serum, vitreous, bile, cerebrospinal, and pericardial fluids in 4 cases of fatal ectopic pregnancy and 40 controls using a quantitative electrochemiluminescence immunoassay designed for living patients. No false-negative cases were identified in any of the analyzed samples in any of the ectopic pregnancy cases. No correlations were found between total HCG levels in postmortem serum and the other tested specimens. The results of this study would suggest that higher HCG in bile, vitreous, pericardial, and cerebrospinal fluids may confirm the existence of ectopic pregnancy and therefore identify other situations in which this hormone is increased, although gestational age cannot be reliably estimated using these values.

2016 - Erratum to: Prognostic relevance of microsatellite instability in pT3N0M0 colon cancer: a population-based study(Intern Emerg Med, Doi:10.1007/s11739-015-1285-6) [Articolo su rivista]
Iachetta, F.; Domati, F.; Reggiani Bonetti, L.; Barresi, V.; Magnani, G.; Marcheselli, L.; Cirilli, C.; Pedroni, M.
abstract

2016 - HOXB13 as an immunohistochemical marker of prostatic origin in metastatic tumors [Articolo su rivista]
Barresi, Valeria; Ieni, Antonio; Cardia, Roberta; Licata, Luana; Vitarelli, Enrica; Reggiani Bonetti, Luca; Tuccari, Giovanni
abstract

HOBX13 is a transcription factor expressed in the normal prostatic glands and overexpressed in prostate cancer. Recent studies suggested that HOXB13 represents a prostate-specific marker in the differential diagnosis between prostatic and urothelial carcinoma. The aim of this study was to analyze and compare the diagnostic value of HOXB13 and prostate-specific antigen (PSA) immunoexpression for the detection of prostatic origin in metastatic tumours. PSA and HOXB13 immunohistochemical expression was assessed in 50 metastatic tumors, including 15 metastases from prostatic adenocarcinoma, 11 from lung adenocarcinoma, 12 from urinary bladder urothelial carcinoma, 11 from colorectal carcinoma, and in 1 from hepatocellular carcinoma. Strong staining for HOXB13 was observed in >75% of neoplastic cells in 15/15 (100%) metastases from prostate cancer. Weak staining in <25% of cells was found in 2/12 (17%) metastases from urothelial carcinoma. PSA immunostaining was detected only in 8 (53%) cases of prostatic origin. The sensitivity and specificity for metastatic prostate cancer were 100% and 94% for HOXB13 and 53% and 100% for PSA. Due to its high sensitivity and specificity, HOXB13 may be included in the pool of prostate-specific markers in metastases showing absent or weak staining for PSA before excluding prostatic derivation.

2016 - HOXB13 expression in metastatic prostate cancer [Articolo su rivista]
Barresi, Valeria; Ieni, Antonio; Reggiani Bonetti, Luca; Tuccari, Giovanni
abstract

Not available

2016 - HR-MAS NMR CONTRIBUTION TO THE COMPREHENSION OF METABOLIC DISORDERS IN INFLAMMATORY AND NEOPLASTIC HYPERPROLIFERATIVE DISEASE [Abstract in Atti di Convegno]
Righi, V.; Schenetti, Luisa; Valentini, A.; REGGIANI BONETTI, Luca; Mucci, Adele
abstract

HR-MAS NMR spectroscopy allows the detailed metabolic analysis of whole biopsy samples, of normal and alterated tissues, to be performed. The NMR spectra constitute a “fingerprint” of the whole metabolome. Correlations between some metabolites and proliferative markers allow gaining insight into the relationship between cellular proliferation and the metabolic changes associated with the presence and tumour aggressiveness. Because metabolites represent the downstream expression of genome, transcriptome and proteome, they can closely reflect the phenotype of an organism at a specific time. Analyzing metabolic differences between normal and perturbed pathways could also provide insight into disease prognosis and diagnosis. Metabolomics could reveal novel cancer biomarkers that might expand our current understanding of this multi-factorial disease. The clinical role of proton NMR in the evaluation of brain tumours, as well as breast and prostate cancer, has been under investigation for several years.[1] During last years our interest was focused on metabolism of colonrectal and brain tissues by using the HR-MAS NMR technique. Some results coming for our research on glioma and linfoma brain tumours and the metabolic correlation existing between colorectal cancer and polyps will be presented.

2016 - Histologic prognostic markers in stage IIA colorectal cancer: A comparative study [Articolo su rivista]
Barresi, Valeria; Reggiani Bonetti, Luca; Ieni, Antonio; Branca, Giovanni; Tuccari, Giovanni
abstract

Objective: pTNM stage IIA colorectal cancer (CRC) is not currently submitted to any adjuvant treatment due to its good prognosis. Nevertheless, a percentage of cases unexpectedly recur. The aim of this study was to assess and compare the prognostic value and inter-observer agreement of a novel histological grading system based on the counting of poorly differentiated clusters (PDC) of cancer cells and that of conventional histological grade, lymphatic, venous and perineural invasion (LVI, VI, PNI), tumour budding (TB) and tumor border configuration in stage IIA CRC.Materials and methods: the afore mentioned histological parameters were assessed in 82 stage IIA CRCs. Inter-observer agreement and correlation with tumour relapse were analyzed by using Fleiss-Cohens weighted K statistics, Fisher exact test and Chi-squared test. The Mantel-Cox log-rank test was applied to assess the strength of association with disease-free interval (DFI).Results: inter-observer agreement was very good/good in the assessment of PDC presence and grade, while it was moderate at best in the evaluation of the other parameters. The presence of PDC, high PDC grade, LVI and TB were significantly associated with disease progression (p < 0.0001; p = 0.0012; p = 0.0308; p = 0.0002) and shorter DFI (p = 0.0001; p < 0.0001; p = 0.0129; p = 0.0008). PDC presence (p < 0.0001) and TB (p = 0.012) were independent prognostic factors in multivariate analysis.Conclusions: our findings suggest that the assessment of PDC may be useful to stratify patients with stage IIA CRC for recurrence risk, and to identify high risk patients who could benefit from adjuvant chemotherapy.

2016 - Loss of alpha-dystroglycan expression in cutaneous melanocytic lesions [Articolo su rivista]
Garcovich, S; Migaldi, Mario; REGGIANI BONETTI, Luca; Capizzi, R; Massimo, L; Boninsegna, A; Arena, V; Cufino, V; Scannone, D; Sgambato, A.
abstract

The dystroglycan complex (DG), originally characterized in muscle and in genetic muscle diseases, links the epithelial cell cytoskeleton to the basement membrane.[1] DG has been shown to be involved in skin morphogenesis and in epithelial carcinogenesis, modulating cell differentiation and adhesion, assembly of the epithelial basement membrane and interactions with the extracellular matrix (ECM).[2] DG comprises an alpha (α-DG) and a beta (β-DG)-subunits, with the α-DG subunit being the extracellular, functional part of the complex and binding several ECM components. Loss of α-DG function has been reported in several epithelial- and neural-derived tumours and correlated with tumour grading, progression and clinical outcome.[3] In the skin, DG is localized at the dermo-epidermal junction and is produced by epidermal keratinocytes and dermal fibroblasts.[4] Tissue expression of the α-DG subunit has not been previously characterized in cutaneous melanocytic nevi and in malignant melanoma. In this study, expression of the DG complex was analysed by Western-blot in cell extracts from human normal melanocytes (hMel) and three melanoma cell lines (BLM, M14, IF6). Furthermore, tissue expression of α-DG was assessed by immunohistochemistry in a panel of archival melanocytic lesions, including melanocytic nevi (n = 20), cutaneous melanoma (n = 51) and melanoma metastases (n = 53). A polyclonal antibody to alpha-DG (clone VIA4-1) from Upstate Biotechnology and a monoclonal antibody to β-DG (clone 43DAG/8D5) from Novocastra were used for the analyses. β-DG molecule was detectable in both melanocytes and melanoma cells displaying a higher expression in IF6 melanoma cells compared to normal melanocytes. On the other hand, a decreased expression of α-DG was observed in all three melanoma cell lines compared to normal melanocytes (Fig. 1). At the tissue level, in benign melanocytic nevi α-DG was expressed by basal melanocytes at the dermo-epidermal junction and by epithelioid melanocytes, organized in nests, in the superficial dermis (Fig. 2a,b). In primary cutaneous melanoma, α-DG expression appeared to be lost in almost all cases. Loss of α-DG expression in melanoma was observed at all Clark invasion levels, both in the epidermis and dermo-epidermal junction as well as in the superficial to deep dermis (Fig. 2c–e). In melanoma metastases, expression of α-DG was evident in 20.8% of cases, being mainly focal and limited to spindle-cell like melanoma cells (Fig. 2f). Previous studies have only characterized the in-vitro expression of DG subunits by melanocytes and melanoma cells, as well as tissue expression of β-DG in melanocytic nevi.[5, 6] We report a differential pattern of α-DG expression between benign and malignant melanocytic tumours, suggesting a peculiar role of the DG complex in melanocyte biology and melanomagenesis. In benign melanocytic nevi, α-DG displayed a consistent expression pattern at the dermo-epidermal junction, where interactions with basal keratinocytes and ECM components are prominent in the epidermal melanin unit. In primary cutaneous melanoma, a loss of α-DG tissue expression was observed across all stages of progression. This finding is compatible with the loss of anchorage to the basement membrane in the early intraepidermal proliferation of transformed melanocytes due to an altered expression of ECM binding proteins.[7] Loss of the α-DG subunit in melanoma is likely to be caused by several post-translational mechanisms, such as an altered glycosylation pattern and/or proteolytic degradation of the membrane complex.[8] Loss of α-DG function results in a disruption of cell-to-ECM interactions and might promote tumour invasion and metastasis. However, the observed expression of α-DG on cells with spindle-cell like morphology in melanoma metastasis suggests that its restoration could favour the implant of cells in metastatic sites, as previously reported in other canc

2016 - Management and therapeutic response of a prostate ductal adenocarcinoma: a still unknown tumour? [Articolo su rivista]
Martorana, Eugenio; Micali, Salvatore; Pirola, Giacomo Maria; REGGIANI BONETTI, Luca; Clo', Vera; Ghaith, A; Bianchi, Giampaolo
abstract

Ductal adenocarcinoma is a rare subtype of prostate cancer with a worse prognosis.Histologically, it is characterized by the presence of tall, pseudostratified columnar epithelium with abundant cytoplasm organized in a papillary or cribriform-papillary pattern. Several clinical differences distinguish this subtype of prostate cancer by the conventional acinar adenocarcinoma: exophytic growth into the prostatic urethra, different clinical presentation, different sites of metastasis and more aggressiveness. The rarity of this tumour forced to base our knowledge on small case series or on individual case reports, and does not help to establish appropriate guidelines. Therefore, the diagnosis of this tumour masks clinical implications that are still not well-understood.We report the case of a 69-year-old Caucasian man with a diagnosis of pure prostate ductal adenocarcinoma that early developed multiple metastases after radical prostatectomy. The patient started hormonal therapy with a fast biochemical and radiologic (positron emission tomography-computed tomography, PET-CT) hormonal escape. Therefore, we took the decision to perform chemotherapy with Taxotere along with prednisolone with a relative stability of prostate-specific antigen (PSA) level, but a new PET-CT scan showed a further progression of the disease. Finally, the patient underwent therapy with Abiraterone acetate that did not stop the cancer progression.No therapeutic options available showed a good control of disease progression. PSA proved to be a poor marker while, on the contrary, PET-CT scan has proved to be particularly useful in the management of the disease progression. More efforts are required to add new knowledge about this tumour and assess what is known until now.

2016 - New lymphatic structures in colon cancer [Articolo su rivista]
Manenti, Antonio; Pavesi, Erica; Farinetti, Alberto; Reggiani Bonetti, Luca
abstract

Non available

2016 - Nonalcoholic steatohepatitis heralding olmesartan-induced sprue-like enteropathy [Articolo su rivista]
Dolci, Marco; Nascimbeni, Fabio; Romagnoli, Dante; REGGIANI BONETTI, Luca; Guaraldi, Giovanni; Mascia, Maria Teresa; Lonardo, Amedeo
abstract

not available

2016 - Poorly differentiated clusters (PDC) in colorectal cancer: What is and ought to be known. [Articolo su rivista]
Reggiani Bonetti, Luca; Barresi, Valeria; Bettelli, Stefania; Domati, Federica; Palmiere, Cristian
abstract

Background: The counting of poorly differentiated clusters of 5 or more cancer cells lacking a gland-like structure in a tumor mass has recently been identified among the histological features predictive of poor prognosis in colorectal cancer. Main body: Poorly differentiated clusters can easily be recognized in the histological sections of colorectal cancer routinely stained with haematoxylin and eosin. Despite some limitations related to specimen fragmentation, counting can also be assessed in endoscopic biopsies. Based on the number of poorly differentiated clusters that appear under a microscopic field of a Ã20 objective lens (i.e., a microscopic field with a major axis of 1 mm), colorectal cancer can be graded into malignancies as follows: tumors with <5 clusters as grade 1, tumors with 5 to 9 clusters as grade 2, and tumors with â¥10 clusters as grade 3. High poorly differentiated cluster counts are significantly associated with peri-neural and lympho-vascular invasion, the presence of nodal metastases or micrometastases, as well as shorter overall and progression free survival to colorectal cancer. Conclusion: The morphological aspects and clinical relevance of poorly differentiated clusters counting in colorectal cancer are discussed in this review.

2016 - Postmortem diagnosis of anaphylaxis in presence of decompositional changes [Articolo su rivista]
Radheshi, Erjon; REGGIANI BONETTI, Luca; Confortini, Annalisa; Silingardi, Enrico; Palmiere, Cristian
abstract

Eosinophil and activated mast cell identification in the spleen combined with mast cell tryptase determination in postmortem serum may diagnose fatal anaphylaxis with a high degree of certainty. Mast cell tryptase measurement and significance in corpses with decompositional changes remains however an issue of controversy. Analogously, immunohistochemistry in corpses with decompositional changes may be influenced by several mechanisms, including protein alteration, antigen diffusion and unspecific antibody binding to disrupted protein structures. The authors present an autopsy case involving a 55-year-old woman who unintentionally received clarithromycin. Due to difficult in administrative procedures, the postmortem examination was performed 96 h after death. Mast cell tryptase was measured in postmortem serum from femoral, aortic and right heart blood. The obtained results were consistent with mast cell activation. Histochemistry (Pagoda Red) and immunohistochemistry (anti-tryptase antibodies) allowed splenic eosinophils and mast cells to be detected. Based on the results of all postmortem investigations, the hypothesis of anaphylaxis following accidental clarithromycin administration was formulated.

2016 - Prognostic relevance of histopathological features in signet ring cell carcinoma of the colorectum [Articolo su rivista]
Barresi, Valeria; Reggiani Bonetti, Luca; Domati, Federica; Baron, Luigi
abstract

Signet ring cell carcinoma (SRCC) is a rare variant of colorectal cancer (CRC), by definition composed of at least 50Â % of neoplastic cells showing signet ring cell morphology. Colorectal SRCC is mainly characterized by aggressive clinical behavior, high pTNM stage and microsatellite instability (MSI). We assessed the prognostic value of several histopathological parameters (histological grade, venous invasion, lymphovascular invasion, MSI, mucin content, tumour budding, pTNM stage) in terms of disease free survival (DFS) and cancer specific survival (CSS) in a series of 32 SRCCs. We confirm that pTNM stage at diagnosis is relevant for predicting DFS and CSS in SRCC. In addition, we show on haematoxylin and eosin or immunohistochemically stained (CD34, podoplanin) sections that venous invasion and lymphovascular invasion are significantly associated with shorter DFS and CSS in SRCC. Notably, venous invasion assed by immunohistochemistry had the highest risk ratio and proved to be the only independent prognostic variable. Finally, we show that histological grade, as assessed on the percentage of formed glands, has prognostic relevance in SRCC as high-grade tumours (<50Â % of glands) had significantly shorter CSS compared to low-grade tumours. This remained an independent variable at multivariate analysis. If our findings are confirmed in further studies, venous invasion as assessed by immunohistochemistry and histological Tgrade might be added to guidelines for SRCC reporting as significant prognostic factors.

2016 - Prognostic relevance of microsatellite instability in pT3N0M0 colon cancer: a population-based study [Articolo su rivista]
Iachetta, Francesco; Domati, Federica; Reggiani-Bonetti, Luca; Barresi, Valeria; Magnani, Giulia; Marcheselli, Luigi; Cirilli, Claudia; Pedroni, Monica
abstract

Although surgery alone represents a curative approach for patients with pT3N0M0 colon cancer, about 15â20Â % of these patients develop a relapse of disease. Microsatellite instability (MSI) is one of the most important molecular markers in colorectal cancer. The aim of this study was to investigate the prognostic relevance of MSI in all pT3N0M0 tumors recorded in the Cancer Registry of the Province of Modenaâ(Northern Italy) within the 2002â2006 period in patients who showed a relapse of disease during the 5-year period of follow-up (59 cases). They were compared to 59 controls similar in clinical and pathological features but with good prognosis. None of the subjects received adjuvant chemotherapy. MSI status was tested using BAT25, BAT26, NR24, and CAT25 fluorescent-labeled mononucleotide markers. The overall prevalence of MSI was 12.7Â % (15 of 118 cases). MSI was detected mainly in mucinous adenocarcinoma (pÂ 

2016 - Prognostic value of the density of Wilms tumour protein 1 (WT-1) positive microvessels in stage IIA colorectal cancer [Articolo su rivista]
Barresi, Valeria; Bonetti, Luca Reggiani; Branca, Giovanni; Vitarelli, Enrica; Ieni, Antonio; Tuccari, Giovanni
abstract

Stage IIA colorectal cancer (CRC) displays 5-year survival rate around 85%; hence it is not currently submitted to any adjuvant treatment. However a percentage of stage IIA CRCs undergoes disease progression and would benefit from additional therapies. In the aim to investigate whether Wilms Tumor-1 (WT-1) expression might represent a prognostic factor in stage IIA CRC, we analyzed its immunohistochemical expression in 90 stage IIA CRCs divided into two groups according to the evidence of disease progression. While WT-1 staining in the neoplastic cells was not significantly associated with any of the clinico-pathological parameters, the density of intra-tumoral microvessels positive for WT-1 (WT-1 MVD) was significantly higher in stage IIA CRCs characterized by disease progression compared to non-recurring tumours and it was significantly and independently associated with shorter disease-free survival. This study is the first to demonstrate that WT-1 MVD may be useful to discriminate high risk patients with stage IIA CRCs and who may benefit from adjuvant treatment. WT-1 expression in the tumor vessels, but not in the vessels of normal colorectal mucosa, suggests its possible role in tumor neo-angiogenesis and it may represent a target for novel anti-angiogenic therapies in stage IIA CRC at high risk of progression.

2016 - Rare metastatic sites of renal cell carcinoma: urethra and spermatic cord [Articolo su rivista]
Pirola, Giacomo M.; Martorana, Eugenio; Fidanza, Francesco A.; Bonetti, Luca Reggiani; Puliatti, Stefano; Bonora, Alessandra; Micali, Salvatore; Bianchi, Giampaolo
abstract

INTRODUCTION: About 20% of patients with a diagnosis of primary renal cell carcinoma (RCC) present directly with metastatic disease. The aim of the present manuscript is to describe, for the first time a RCC metastasis located to the distal female urethra and to present an update on metastatic locations to the male spermatic cord.MATERIALS AND METHODS: We report two cases of rare RCC metastases. The first concerns a 92-year-old female patient who came to our attention for recurrent urethral bleeding, which was initially believed to be secondary to urethral mucosal ectropion. Pathology demonstrated a RCC metastasis. The second concerns a 67-year-old male patient with a previous history of RCC who came to our attention for the finding of palpable, mobile and indolent right inguinal lump. Given the past history of malignancy, it was excised and revealed to be a RCC metastasis.DISCUSSION: In the first case, the pathologic specimen allowed the detection of an unknown renal tumor, whereas in the second, the previous neoplastic history of the patient has led clinicians to focus on a possible neoplastic recurrence, perform a correct excision of the node, and begin an early systemic therapy.CONCLUSIONS: These cases are emblematic of possible unexpected RCC metastasis. These findings should be taken into account in order to clarify the differential diagnosis and to address these patients to a correct therapeutic course.

2016 - The lymphatic anatomy of the mesorectum helps to understand rectal cancer recurrence [Articolo su rivista]
Manenti, Antonio; Bonetti, Luca Reggiani; Farinetti, Alberto; Sani, Francesco
abstract

Not available

2016 - Therapeutic and recreational methadone cardiotoxicity [Articolo su rivista]
Lusetti, M; Licata, Manuela; Silingardi, Enrico; Reggiani Bonetti, L; Palmiere, C.
abstract

Several classes of drugs have been associated with an increased risk of cardiovascular disease and occurrence of arrhythmias potentially involved in sudden deaths in chronic users even at therapeutic doses. The study presented herein focuses on pathological changes involving the heart possibly due to methadone use. 60 cases were included in the study in total and were divided into three groups (therapeutic methadone users: 20 cases, recreational methadone users: 20 cases, and sudden death group in subjects who had never taken methadone: 20 cases). Autopsies, histology, biochemistry and toxicology were performed in all cases. Macroscopic and microscopic investigation results in therapeutic methadone users were similar to those observed in sudden, unexpected deaths in non-methadone users. In recreational methadone consumers, macroscopic and microscopic examination of the heart failed to provide results consistent with acute or chronic myocardial or coronary damage, thereby corroborating the hypothesis of death most likely following respiratory depression.

2015 - A comparative analysis between laparoscopy and open colectomy: assessment of perioperative and oncological outcomes [Articolo su rivista]
Farinetti, Alberto; Bonetti, Luca Reggiani; Migaldi, Mario; Mattioli, Anna Vittoria; Pennella, Sonia; Muratori, Simone; Rossi, Aldo
abstract

AIM: Aim of the study was to compare two groups of patients affected by colorectal adenocarcinoma that underwent to open colectomy and laparoscopic colectomy respectively, highlighting the advantage and problems. MATERIAL OF STUDY: This is a retrospective analysis. Between January 2003 and December 2006, 54 patients who underwent colectomy were recruited. Of these, 26 patients underwent open colectomy, and 28 laparoscopy. RESULTS: For open colectomy the average duration of surgery was 177.9 minutes (surgical time) and 280.4 minutes (time of operating room) with a minimum of 110 and a maximum of 360 minutes, with significant differences according to type of surgery performed and the patient’s clinical history. For laparoscopy the average duration was 293 minutes, (range 135 - 520), with significant differences depending on the portion of the intestinal tract removed. DISCUSSION: The comparison of two different surgical techniques, laparoscopic and open colectomy revealed some differences. The duration of the resection was greater for laparoscopy when compared to the traditional technique. CONCLUSIONS: Both approaches are technically feasible, safe and oncologically correct. Laparoscopic technique shows a much more favorable outcome in terms of pain, absence of extensive scarring, the incidence of incisional hernias and hospital stay -surgery compared with surgery laparotomy.

2015 - Aberrant DNA methylation profiles of inherited and sporadic colorectal cancer [Articolo su rivista]
Sahnane, Nora; Magnoli, Francesca; Bernasconi, Barbara; Tibiletti, Maria Grazia; Romualdi, Chiara; Pedroni, Monica; De Leon, Maurizio Ponz; Magnani, Giulia; Reggiani-Bonetti, Luca; Bertario, Lucio; Signoroni, Stefano; Capella, Carlo; Sessa, Fausto; Furlan, Daniela
abstract

Background: Aberrant DNA methylation has been widely investigated in sporadic colorectal carcinomas (CRCs), and extensive work has been performed to characterize different methylation profiles of CRC. Less information is available about the role of epigenetics in hereditary CRC and about the possible clinical use of epigenetic biomarkers in CRC, regardless of the etiopathogenesis. Long interspersed nucleotide element 1 (LINE-1) hypomethylation and gene-specific hypermethylation of 38 promoters were analyzed in multicenter series of 220 CRCs including 71 Lynch (Lynch colorectal cancer with microsatellite instability (LS-MSI)), 23 CRCs of patients under 40 years in which the main inherited CRC syndromes had been excluded (early-onset colorectal cancer with microsatellite stability (EO-MSS)), and 126 sporadic CRCs, comprising 28 cases with microsatellite instability (S-MSI) and 98 that were microsatellite stable (S-MSS). All tumor methylation patterns were integrated with clinicopathological and genetic characteristics, namely chromosomal instability (CIN), TP53 loss, BRAF, and KRAS mutations. Results: LS-MSI mainly showed absence of extensive DNA hypo-and hypermethylation. LINE-1 hypomethylation was observed in a subset of LS-MSI that were associated with the worse prognosis. Genetically, they commonly displayed G:A transition in the KRAS gene and absence of a CIN phenotype and of TP53 loss. S-MSI exhibited a specific epigenetic profile showing low rates of LINE-1 hypomethylation and extensive gene hypermethylation. S-MSI were mainly characterized by MLH1 methylation, BRAF mutation, and absence of a CIN phenotype and of TP53 loss. By contrast, S-MSS showed a high frequency of LINE-1 hypomethylation and of CIN, and they were associated with a worse prognosis. EO-MSS were a genetically and epigenetically heterogeneous group of CRCs. Like LS-MSI, some EO-MSS displayed low rates of DNA hypo-or hypermethylation and frequent G:A transitions in the KRAS gene, suggesting that a genetic syndrome might still be unrevealed in these patients. By contrast, some EO-MSS showed similar features to those observed in S-MSS, such as LINE-1 hypomethylation, CIN, and TP53 deletion. In all four classes, hypermethylation of ESR1, GATA5, and WT1 was very common. Conclusions: Aberrant DNA methylation analysis allows the identification of different subsets of CRCs. This study confirms the potential utility of methylation tests for early detection of CRC and suggests that LINE-1 hypomethylation may be a useful prognostic marker in both sporadic and inherited CRCs.

2015 - Advantages of single-puncture transperineal saturation biopsy of prostate: analysis of outcomes in 125 patients using our scheme [Articolo su rivista]
Martorana, Eugenio; Micali, Salvatore; Ghaith, Ahmed; REGGIANI BONETTI, Luca; Sighinolfi, Maria Chiara; Galli, Riccardo; Paterlini, Maurizio; Bianchi, Giampaolo
abstract

PURPOSE: Stereotactic biopsy has improved prostate cancer detection. Although the new approach is superior, standard procedure is still useful in a cohort of patients in whom MRI is not available. The standard saturation biopsy technique is still debatable. We describe our technique and analyze its outcomes. MATERIALS AND METHODS: One hundred twenty-five patients underwent saturation biopsy through a single transperineal access. Mean age was 64.73 year, mean PSA was 9.49 ng/ml, mean PSA density was 0.184, and mean prostate volume was 57.95 g. All patients underwent at least one previous prostatic biopsy: 24.8% of cases had diagnosis of atypical small acinar proliferation, 39.2% of cases had multifocal high-grade prostatic intraepithelial neoplasia, and 36% of cases had inflammation or benign prostatic hyperplasia. RESULTS: The detection rate was 38.4%. Prostate cancer occurred in 61.3% of patients with previous ASAP (p < 0.007). Cancer detection rate decreased with increasing number of previous biopsy and with increasing prostate volume (p < 0.001) and increased with increasing PSA density (p = 0.03). No major complications were reported. CONCLUSION: The traditional saturation biopsy may be useful when targeted biopsy cannot be used. Our technique is accurate for cancer detection. It can offer some advantages in comparison with other approaches.

2015 - Assessment of freezing effects and diagnostic potential of BioBank healthy and neoplastic breast tissues through HR-MAS NMR spectroscopy [Articolo su rivista]
Righi, Valeria; Schenetti, Luisa; Maiorana, Antonino; Libertini, Emanuela; Bettelli, Stefania Raffaella; Reggiani Bonetti, Luca; Mucci, Adele
abstract

HR-MAS NMR spectroscopy was employed to monitor the metabolic profiles of Modena BioBank breast samples over 1 year of freezing at -80 C. The study includes 22 adult female patients living in Modena and its hinterland, who underwent total mastectomy or quadrantectomy in 2011–2012. Variations occur, especially affecting phosphocholine (PC) and choline. This is not a trivial finding, since many studies base the distinction between neoplastic and healthy tissues or the assessment of tumor grade on the analysis of choline metabolites. Despite the changes observed, we established that the diagnostic power of the HR-MAS NMR spectra of frozen samples is preserved, at least as far as the distinction between neoplastic and healthy samples is concerned. Lactate (Lac), PC, phosphoethanolamine (PE), taurine (Tau), myo-inositol (Myo) and glucose (Glc) are biomarkers that can be used to distinguish healthy from neoplastic tissues, whereas some metabolite ratios, such as Lac ? PE ? Tau/Glc ? Myo, seem to have even higher discrimination potential.

2015 - Cardiac Toxicity in Selective Serotonin Reuptake Inhibitor Users [Articolo su rivista]
Lusetti, M; Licata, Manuela; Silingardi, Enrico; Reggiani Bonetti, L; Palmiere, C.
abstract

Several classes of recreational and prescription drugs have been associated with an increased risk of cardiovascular disease and the occurrence of arrhythmias, which may be involved in sudden deaths in chronic users even at therapeutic doses. The study presented herein focuses on pathological changes involving the heart, which may be caused by selective serotonin reuptake inhibitor use and their possible role in the occurrence of sudden cardiac death. A total of 40 cases were included in the study and were divided evenly into 2 groups: 20 cases of patients treated with selective serotonin reuptake inhibitors and 20 cases of sudden deaths involving patients void of any drug treatment. The first group included 16 patients treated with citalopram and 4 with sertraline. Autopsies, histology, biochemistry, and toxicology were performed in all cases. Pathological changes in selective serotonin reuptake inhibitor users consisted of various degrees of interstitial and perivascular fibrosis as well as a small degree of perineural fibrosis within the myocardium of the left ventricle. Within the limits of the small number of investigated cases, the results of this study seem to confirm former observations on this topic, suggesting that selective serotonin reuptake inhibitors may play a potential, causative role in the pathogenesis of sudden deaths in chronic users even at therapeutic concentrations.

2015 - Cerebrospinal fluid PCR analysis and biochemistry in bodies with severe decomposition [Articolo su rivista]
Palmiere, Cristian; Vanhaebost, Jessica; Ventura, Francesco; Bonsignore, Alessandro; Bonetti, Luca Reggiani
abstract

The aim of this study was to assess whether Neisseria meningitidis, Listeria monocytogenes, Streptococcus pneumoniae and Haemophilus influenzae can be identified using the polymerase chain reaction technique in the cerebrospinal fluid of severely decomposed bodies with known, noninfectious causes of death or whether postmortem changes can lead to false positive results and thus erroneous diagnostic information. Biochemical investigations, postmortem bacteriology and real-time polymerase chain reaction analysis in cerebrospinal fluid were performed in a series of medico-legal autopsies that included noninfectious causes of death with decomposition, bacterial meningitis without decomposition, bacterial meningitis with decomposition, low respiratory tract infections with decomposition and abdominal infections with decomposition. In noninfectious causes of death with decomposition, postmortem investigations failed to reveal results consistent with generalized inflammation or bacterial infections at the time of death. Real-time polymerase chain reaction analysis in cerebrospinal fluid did not identify the studied bacteria in any of these cases. The results of this study highlight the usefulness of molecular approaches in bacteriology as well as the use of alternative biological samples in postmortem biochemistry in order to obtain suitable information even in corpses with severe decompositional changes.

2015 - Correlation between microsatellite instability status and grading assessed by the counting of poorly differentiated clusters in colorectal cancer [Articolo su rivista]
Barresi, Valeria; Bonetti, Luca Reggiani
abstract

Not available

2015 - Histological grading in colorectal cancer: New insights and perspectives [Articolo su rivista]
Barresi, Valeria; Bonetti, Luca Reggiani; Leni, Antonio; Caruso, Rosario Alberto; Tuccari, Giovanni
abstract

Poor histological differentiation is currently considered among the adverse histopathological factors associated with unfavourable clinical course of colorectal carcinoma (CRC). At present, the histological grade of CRC is assessed based on the percentage of glandular differentiation in the tumor according to the World Health Organization (WHO) criteria. However the prognostic value of the WHO grading system is limited by significant inter-observer variability in its assessment. In addition, the prognostic significance of WHO grading seems to depend on the microsatellite instability (MSI) status of the tumor. Finally, this grading does not apply to rarer histotypes of colorectal adenocarcinomas, such as the micropapillary, medullary, mucinous and signet ring cell variants. Recently a novel grading system based on the counting of clusters of five or more cells lacking a glandular structure (poorly differentiated clusters) and set in the tumor stroma or at invasive edge has been proposed in CRC. There is evidence that grading based on poorly differentiated clusters (PDC) is more reproducible and has more robust prognostic significance compared to WHO grading in CRC. In the present review we discuss the morphological features, criteria for the assessment, prognostic significance and correlation with biomolecular profiles of grading based on PDC counting in CRC.

2015 - Human cytomegalovirus induced pseudotumor of upper gastrointestinal tract mucosa: Effects of long-term chronic disease? [Articolo su rivista]
Reggiani Bonetti, Luca; Barresi, Valeria; Bertani, Angela; Maccio, Livia; Palmiere, Cristian
abstract

Human cytomegalovirus-induced lesions resembling malignancies have been described in the gastrointestinal tract and include ulcerated or exophytic large masses. The aim of this study was to review the cases registered in the databases of two academic hospitals and formulate a hypothesis concerning the pathogenic mechanisms responsible for cytomegalovirus-induced pseudotumor development. All the diagnoses of human cytomegalovirus infections of the upper gastrointestinal tract recorded from 1991 to 2013 were reviewed. Cases of mucosal alterations misdiagnosed endoscopically as malignancies were selected. Large ulcers occurring in the stomach (three cases) and an irregular exophytic mass at the gastro-jejunal anastomosis were misdiagnosed endoscopically as malignancies (4 cases out of 53). Histologically, all lesions reflected hyperplastic mucosal changes with a prevalence of epithelial and stroma infected cells, without signs of cell atypia. The hypothesis presented is that the development of human cytomegalovirus-induced pseudotumors may be the morphological expression of chronic mucosa damage underlying long-term infection.

2015 - KRAS, NRAS, BRAF mutations and high counts of poorly differentiated clusters of neoplastic cells in colorectal cancer: Observational analysis of 175 cases [Articolo su rivista]
Barresi, Valeria; Bonetti, Luca Reggiani; Bettelli, Stefania
abstract

A novel grading system based on the counting of poorly differentiated clusters (PDC) of neoplastic cells at the invasive margin and in the tumour stroma was recently introduced among the histological parameters predictive of adverse clinical outcome in colorectal cancer (CRC). The aim of this study was to correlate the histological grade based on PDC and the mutational status of KRAS, NRAS and BRAF genes in 175 consecutive CRCs. The highest PDC count under the objective lens of a x20 microscopic field in each tumour was considered for grading assessment, so that PDC counts <5, 5-9 and â¥10 PDCs were defined grade 1, grade 2 and grade 3, respectively. Hotspots mutations were identified using the MassArray platform. Overall, there were 42 (24%) mutated tumours. Mutational status was significantly associated with high pT stage (p = 0.0021), advanced pTNM stage (p = 0.0018), nodal metastases (p = 0.006), tumour budding (p = 0.022) and high PDC grade (p = 0.0022). KRAS mutations were significantly associated with PDC grade (p = 0.0379), while BRAF mutations were associated with PDC-G3 although statistical significance was not reached. No significant associations were found between NRAS and PDC. The significant association between mutated KRAS and PDC grade suggests that KRAS mutations may be involved in the formation of PDC.

2015 - Malignant metastasizing solitary fibrous tumors of the liver: A report of three cases [Articolo su rivista]
Maccio, Livia; Bonetti, Luca Reggiani; Siopis, Elena; Palmiere, Cristian
abstract

Solitary fibrous tumors are rare neoplasms of mesenchymal origin that have been reported in various other extrathoracic sites, including the liver. We present a case series of three malignant solitary fibrous tumors of the liver, occurring in two women 74 and 80 years old and one 65-year-old man. No clinical features were predictive of malignancy except the large sizes and synchronous presence of lung metastases in two of the three cases. Histological examinations revealed the presence of high pleomorphic cellularity with nuclear atypia, necrosis and high mitotic ratios. All patients died of disease progression.

2015 - Molecular features and methylation status in early onset (< 40 years) colorectal cancer: a population based, case-control study [Articolo su rivista]
Magnani, Giulia; Furlan, Daniela; Sahnane, Nora; Reggiani Bonetti, Luca; Domati, Federica; Pedroni, Monica
abstract

Colorectal cancer is usually considered a disease of the elderly. However, a small fraction of patients develops colorectal cancer earlier. The aim of our study was to define the frequency of known hereditary colorectal syndromes and to characterise genetic and epigenetic features of early nonhereditary tumors. Thirty-three patients ≤40 years with diagnosis of colorectal cancer and 41 patients with disease at >60 years of age were investigated for MSI, Mismatch Repair proteins expression, KRAS and BRAF mutations, hypermethylation, and LINE-1 hypomethylation. Detection of germline mutations was performed in Mismatch Repair, APC and MUTYH genes. Early onset colorectal cancer showed a high incidence of hereditary forms (18%). KRAS mutations were detected in 36% of early nonhereditary tumors. Early onset colorectal cancer disclosed an average number of methylated genes significantly lower when compared to the controls (). Finally both of the two groups were highly methylated in ESR1, GATA5, and WT1 genes and were similar for LINE-1 hypomethylation. The genetic make-up of carcinomas differs from young to elderly patients. Early onset tumors showed more frequently a constitutional defective of Mismatch Repair System and a minor number of methylated genes. Hypermethylation of ESR1, GATA5, and WT1 genes suggests possible markers in the earlier diagnosis of colorectal tumorigenesis.

2015 - Pathological changes in anabolic androgenic steroid users [Articolo su rivista]
Lusetti, M; Licata, Manuela; Silingardi, Enrico; Reggiani Bonetti, L; Palmiere, C.
abstract

Several classes of recreational and prescription drugs have additional effects on the heart and vasculature, which may significantly contribute to morbidity and mortality in chronic users. The study presented herein focuses on pathological changes involving the heart possibly due to anabolic androgenic steroid use. The role these hormones may play in their occurrence of sudden cardiac death is also investigated. 98 medico-legal cases including 6 anabolic androgenic steroid users were retrospectively reviewed. Autopsies, histology, immunohistochemistry, biochemistry and toxicology were performed in all cases. Pathological changes consisted of various degrees of interstitial and perivascular fibrosis as well as fibroadipous metaplasia and perineural fibrosis within the myocardium of the left ventricle. Within the limits of the small number of investigated cases, our results appear to confirm former observations on this topic and suggest anabolic androgenic steroid's potential causative role in the pathogenesis of sudden cardiac deaths in chronic users

2015 - Prognostic significance of grading based on the counting of poorly differentiated clusters in colorectal mucinous adenocarcinoma [Articolo su rivista]
Barresi, Valeria; Reggiani Bonetti, Luca; Ieni, Antonio; Domati, Federica; Tuccari, Giovanni
abstract

Mucinous adenocarcinoma (MAC) of the colon and rectum is a histological entity with still indefinite prognostic significance. Although it was previously designated as poorly differentiated by convention, the most recent World Health Organization guidelines indicate that the level of maturation of the epithelium determines differentiation in MAC and that microsatellite instability status should be taken into account for its histological grading. Nonetheless, precise criteria for grading are not provided, and the prognostic value of histological grading in MAC still remains unclear. In the present study we aimed to investigate the prognostic value of a grading system based on the counting of poorly differentiated clusters (PDC) of neoplastic cells in 108 colorectal MACs and to compare its reproducibility and significance with those of a grading system based on glandular differentiation. We found that PDC grade was more reproducible and significantly associated with disease progression (P =.0089) as well as with death from colorectal cancer (P =.0035) in our MACs, as compared to the grade based on glandular differentiation, which was not associated with any of the clinicopathologic variables. Moreover, PDC grade emerged as a significant, independent prognostic factor of recurrence-free survival (P =.0198) and cancer-specific survival (P =.0293) in MAC. Interestingly, the prognostic value of PDC grade was unaltered following incorporation of mismatch repair system status in grading. In conclusion, we demonstrated for the first time that PDC grading is feasible, reproducible, and prognostically relevant in MAC, which may support its use in routine practice.

2015 - ROBOTIC DISTAL PANCREATECTOMY FOR PANCREATIC LEIOMYOSARCOMA [Articolo su rivista]
Tarantino, G; Ballarin, R; Bonetti, L R; Di Benedetto, F
abstract

Objective: Primary leiomyosarcoma of the pancreas (PLMS) is an extremely rare tumour. Herein we report the first case of robotic distal pancreatectomy (DP) ever performed so far. Patiens and Methods: A 73-year-old woman was referred to our Institution after a diagnosis of pancreatic neoplasm was made. She underwent robotic assisted DP. Results: The patient was discharged to home in the fourth post-operative day. Immunohistochemical staining was positive for smooth muscle markers, with negative expression of epithelial and neural markers, thus confirming the diagnosis of high grade PLMS. Conclusions: Robotic DP combined good oncological results together with the advantages of minimally access surgery.

2015 - Radiotherapy-induced mesorectum alterations: histological evaluation of 90 consecutive cases. [Articolo su rivista]
REGGIANI BONETTI, Luca; Domati, Federica; Farinetti, Alberto; Migaldi, Mario; Manenti, Antonio
abstract

Abstract Objective. In order to identify the radiotherapy-induced histological modifications in the mesorectum, we reviewed the surgical specimens of 90 rectal resections comprehensive of the total mesorectal excision (23 cases radiologically classified as cT2N0M0 and 67 as cT3N0M0). All patients were preoperative treated with radiotherapy: 20 with 50 Gy, 20 with 20 Gy and 50 Gy irradiation associated to FOLFOX scheme chemotherapy. Material and methods. Routine hematoxylin and eosin stained serial slides at 5 mm of intervals were obtained from surgical specimens and included the tumor site and the adjacent irradiated mucosa, the submucosa and the muscular layers of the rectal wall and the mesorectal adipose tissue, completely removed until to the mesorectal fascia. Ten subjects (eight cT2N0M0 and two cT3N0M0), who did not received preoperative oncological treatments were adopted as controls. Results. Histologically, examination revealed fibrosis of the adipose tissue in 86 cases (95%), vascular damage including vasculities and fibrotic thickening wall of arteries and veins in 46 cases (51%), sclero-hyalinosis of lymph nodes with pericapsular fibrosis in 22 cases (23%) and perineural deposition of fibrosis in 12 (13%). These findings were ubiquitously observed in the whole mesorectum. Fibrosis of the adipose tissue and vasculitis were mainly associated to the combination of 50 Gy radiations plus chemotherapy (p < 0.05). Conclusion. The detection of histopathological alterations in the mesorectum can give reason of the well-known postoperative complications and long-term sequels.

2015 - The importance of inflammation in the search of prostate cancer biomarkers [Poster]
Bergamini, S; Bellei, E; Monari, E; Cuoghi, A; Reggiani Bonetti, L; Borelli, F; Sighinolfi, C; Bianchi, G; Ozben, T; Tomasi, A
abstract

The importance of inflammation in the search of prostate cancer biomarkers by proteomics.

2015 - The perception of health-related quality of life in colon cancer patients during chemotherapy: differences between men and women [Articolo su rivista]
Domati, Federica; Luppi, Gabriele; Reggiani-Bonetti, Luca; Zironi, Sandra; Depenni, Roberta; Fontana, Annalisa; Gelsomino, Fabio; de Leon, Maurizio Ponz
abstract

There is a need for more information on the quality of life (QoL) in patients undergoing chemotherapy. We wanted to investigate the perception of health status in colon cancer patients before, 3 and 6Â months after chemotherapy. A secondary purpose was to assess the different perceptions of QoL between men and women during and after adjuvant or palliative therapy. We investigated 100 patients throughout chemotherapy for colon cancer. Data were collected through the SF-36 questionnaire. The score of all variables analyzed in the study group was lower than in the control group, which indicates a lower performance status, more marked in the female sex. Patients were then subdivided by the state of disease (localized or metastatic) and the variables, were evaluated before, 3 and 6Â months after therapy. In patients treated with adjuvant treatment, there was a worsening of the performance status, followed by an increase after 6Â months. We found that after 3Â months of therapy, affected male patients perceived more limitations in carrying out their work, other daily activities and social relationships, owing to both their emotional state and their physical health. In metastatic patients the values of the eight variables decreased dramatically after 6Â months, indicating a worsening of the QoL. In patients who received adjuvant treatment there was a certain worsening of the health status at 3Â months, followed by a general improvement after 6Â months. This improvement was not observed in patients undergoing palliative therapy. Several differences were observed between men and women in performance status after treatment.

2014 - ASSESSMENT OF FREEZING EFFECTS AND DIAGNOSTIC POTENTIAL OF BIOBANK HEALTHY AND NEOPLASTIC BREAST TISSUES THROUGH HR-MAS NMR SPECTROSCOPY [Abstract in Atti di Convegno]
Mucci, Adele; Schenetti, Luisa; Maiorana, Antonino; Bettelli, Stefania Raffaella; REGGIANI BONETTI, Luca; V., Righi
abstract

HR-MAS NMR spectroscopy was employed to monitor the metabolic profiles of Modena BioBank breast samples over one year of freezing at -80 °C. The study includes 22 adult female patients living in Modena and its hinterland, who underwent total mastectomy or quadrantectomy in 2011 - 2012. Variations occur, especially affecting phosphocholine and choline. This is not a trivial finding, since many studies base the distinction between neoplastic and healthy tissues or the assessment of tumor grade on the analysis of choline metabolites [1,2]. Despite the changes observed, we established that the diagnostic power of the HR-MAS NMR spectra of frozen samples is preserved, at least as far as the distinction between neoplastic and healthy samples is concerned. Lactate, phosphocholine, phosphoethanolamine, taurine, myo-inositol and glucose are biomarkers that can be used to distinguish healthy from neoplastic tissues, whereas some metabolite ratios, such as Lac+PE+Tau/Glc+Myo, seem to have even higher discrimination potential. Fig. 1. Scores plot of PCA on neoplastic (crosses) vs healthy (triangles) samples. Loadings profiles (right) of PC2 and PC3. References [1]

2014 - An unusual case of familial adenomatous polyposis with very early symptom occurrence [Articolo su rivista]
PONZ DE LEON, Maurizio; Bianchini, Maria Anastasia; REGGIANI BONETTI, Luca; Pedroni, Monica; Di Gregorio, Carmela; Merighi, Alberto; Rossi, Giuseppina; Magnani, Giulia; Domati, Federica; Cacciari, Alfredo
abstract

We report the clinical case of a patient who showed an "accelerated" form of polyposis, with development of major lesions within the first decade of life. The patient belongs to a familial adenomatous polyposis family-already described in 2001-featured by profuse polyposis at an early age of onset and desmoid tumors in the majority of affected individuals (of both sexes). The family was characterized by an uncommon mutation of the APC gene (c.4391_4700del310insCACCTACTGCTGAAA, previously defined as c.4394ins15del310) consisting in a large deletion of 310 bp at codon 1,464 with duplication of the breakpoint leading to a stop codon at position 1,575. The proband was affected by desmoids tumors at the age of 3 years. In the same year (2004) numerous polyps in the large bowel and a hepatoblastoma developed. After several months new desmoids appeared in the surgical scar. In 2010, at age 9, the patient was operated of total colectomy and endorectal pull-through of the small intestine owing to profuse colorectal adenomatosis. New desmoids developed in 2011 and 2012, and required chemotherapy. Further analysis of the APC gene in the proband revealed several polymorphisms. One of these (c.398A>G) had not been previously reported, nor was present in two other affected members of the family. The clinical case, and the practical implications for therapy, are discussed according to the most recent theories of colorectal cancer development. Long-term treatment with Cox-2 inhibitors might represent a good option for this patient.

2014 - Clinical outcome of low- and high-risk malignant colorectal polyps: results of a population-based study and meta-analysis of the available literature [Articolo su rivista]
Di Gregorio, Carmela; Bonetti, Luca Reggiani; DE GAETANI, Carmela; Pedroni, Monica; Kaleci, Shaniko; PONZ DE LEON, Maurizio
abstract

Some histological features of malignant polyps have been used to classify patients into low- and high-risk groups. This study proposed to evaluate the impact of this classification on the clinical outcome of patients with malignant polyps. Through the Colorectal Cancer Registry, 105 patients with endoscopically removed malignant polyps were selected. The presence of one of the following histological features defined malignant polyps as high-risk: infiltrated resection-margin, poorly differentiated carcinoma, lymphatic/vascular invasion and tumour budding and depth of submucosal invasion. Available literature was reviewed by applying a similar classification. Most of the malignant polyps were pedunculated and were localized in the left colon. Fifty-five malignant polyps were classified as low-risk lesions and 50 as high-risk. None of the patients at low-risk died of colorectal cancer. Of the patients at high-risk, three died of cancer; all three cases showed lymphatic/vascular invasion. Review of the literature reveals that an unfavourable clinical outcome is significantly more prevalent in the high-risk compared with the low-risk group (p > 0.005). Moreover, all histological risk factors show a specific predictive value of clinical adverse outcome. Our study and the pooled data analysis confirmed the usefulness of the subdivision into low- and high-risk malignant polyps for management of patients with endoscopically removed colorectal carcinoma.

2014 - Discordance rate of her2 status in primary gastric carcinomas and synchronous lymph node metastases: A multicenter retrospective analysis [Articolo su rivista]
Ieni, Antonio; Barresi, Valeria; Caltabiano, Rosario; Caleo, Alessia; Bonetti, Luca Reggiani; Lanzafame, Salvatore; Zeppa, Pio; Caruso, Rosario Alberto; Tuccari, Giovanni
abstract

Background: The assessment of human epidermal growth factor receptor 2 (HER2) gene amplification is essential in order to identify those patients affected by advanced gastric cancer who may benefit from Trastuzumab targeted therapy. Materials and Methods: With the aim to investigate the concordance rate in HER2 status between primary gastric carcinoma (GC) and synchronous lymphnode metastases, we investigated HER2 status in a cohort of 108 surgical formalin-fixed paraffin-embedded specimens of GC and matched synchronous metastatic lymph nodes collected from three different units of Anatomic Pathology in southern of Italy. Fleiss-Cohen weighted k statistics were used to assess the concordance rate of HER2 status. Results: HER2 amplification was observed in 17% of primary GCs and the overall concordance rate with corresponding nodal metastases was 90.74%. Changes in HER2 status between primary GC and matched synchronous metastases were evidenced in 10 (9.26%) cases. Of these, 6 cases were HER2 amplified in the primary GC and not amplified in the metastases, while 4 were HER2 not amplified in the primary tumour and amplified in the lymph node metastases. Conclusions: Although at present the simultaneous determination of HER2 in advanced gastric cancer and corresponding metastatic lymph nodes is not mandatory, the possibility that the synchronous metastases of GC have a different HER2 status from that of the primary tumour is of remarkable significance; Indeed this may have influence on the therapeutic management and prognosis of the patients.

2014 - Double heterozygosity for BRCA1 and hMLH1 gene mutations in a 46-year-old woman with five primary tumors [Articolo su rivista]
Pedroni, Monica; DI GREGORIO, Carmela; Cortesi, Laura; REGGIANI BONETTI, Luca; Magnani, Giulia; Simone, Maria Luisa; Medici, Veronica; PRIORE OLIVA, Claudio; Marino, Marco; PONZ DE LEON, Maurizio
abstract

Germline mutations in BRCA1 and BRCA2 genes predispose to hereditary breast cancer, whereas carriers of mutations in any of the mismatch repair genes (MMR; hMLH1, hMSH2, hMSH6, hPMS2) are highly susceptible to Lynch syndrome. In the present study, we describe a woman affected by unilateral breast cancer at the age of 35 years. After 4 years, during the follow-up she developed synchronous (and asymptomatic) endometrial cancer, ovarian carcinoma and renal clear cell carcinoma. After 7 years (at age 46), the patient developed an infiltrating carcinoma of the contralateral breast and died in a few months of metastatic disease. Initial investigations led to the detection of a constitutional mutation in the BRCA1 gene. The extended genealogical tree disclosed a suspected history of colorectal carcinoma in the maternal branch. Endometrial cancer of the proband was investigated for microsatellite instability (MSI) and immunohistochemical expression of MLH1, MSH2 and MSH6 proteins. An high MSI status and lack of expression of MLH1 protein were detected. hMLH1 gene sequencing revealed the presence of a constitutional mutation, which was also found in the mother of the proband. Loss of the wild-type hMLH1 allele was detected in both breast tumors, thus suggesting that the MMR defect contributed to the development of the breast cancer.

2014 - Expression of CD133 Correlates with Tumor Stage, Lymph Node Metastasis and Recurrence in Oral Squamous Cell Carcinoma [Articolo su rivista]
Reggiani Bonetti, L; Migaldi, Mario; Boninsegna, A; Fanali, C; Farina, M; Chiarini, Luigi; Anesi, Alexandre; Cittadini, A; Leocata, P; Maccio, L; Sgambato, A.
abstract

Objective: CD133 is a pentaspan membrane protein expressed by the so-called cancer stem cells (CSC) in the majority of human cancers. The aim of this study was to analyze CD133 expression levels in specimens of oral cancer and to evaluate its relation with disease evolution. Methods: Expression of the CD133 protein was evaluated by immunostaining in a series of oral squamous cell carcinoma (OSCC) and its relation with traditional prognostic indicators and the clinical outcome of patients was analyzed. Results: CD133 expression was highly variable amongst different samples with a median percentage of positive cells of 5 (range 0 - 80; mean = 11) and CD133 staining was not detectable in tumor cells in 29 (43%) cases. No correlation was observed with age at diagnosis, gender and grading while a significant correlation was observed with tumor stage. Kaplan-Meier curves of disease-free survival displayed a significant separation between the negative and positive groups of patients (p = 0.001 by log-rank test) but CD133 staining did not confirm to be an independent predictor of clinical outcome in a multivariate analyses. Conclusion: Expression of CD133 was detectable in the majority of OSCC samples and correlated significantly with tumor stage and the clinical outcome of patients in terms of disease-free survival. Further studies are warranted on a larger series of cases to elucidate the role of CD133 in the development and progression of OSCC and its suitability as a prognostic biomarker.

2014 - Histologic grading based on counting poorly differentiated clusters in preoperative biopsy predicts nodal involvement and pTNM stage in colorectal cancer patients [Articolo su rivista]
Barresi, Valeria; Bonetti, Luca Reggiani; Ieni, Antonio; Branca, Giovanni; Baron, Luigi; Tuccari, Giovanni
abstract

Histologic grading is commonly assessed in colorectal cancer preoperative biopsies. Nevertheless, its clinical impact is limited by low interobserver reproducibility and poor concordance with grading found in the final resection specimen. In the present study, we aimed to investigate the reproducibility, accuracy, and predictive value on lymph node status or pTNM stage of a novel grading system based on the number of poorly differentiated clusters in colorectal cancer preoperative endoscopic biopsies. Grading based on counting poorly differentiated clusters was assessed in 163 colorectal cancer endoscopic biopsies and corresponding surgical specimens. With this system, 152 biopsies could be graded with good interobserver agreement (Îº = 0.735). In comparison with the surgical specimens, 75% of colorectal cancers were correctly graded in the biopsy, and 81% of poorly differentiated colorectal cancers were identified at initial biopsy. High poorly differentiated clusters grade in the biopsy was significantly associated with nodal metastasis, high pTNM stage (P <.0001), or histologic features suggestive of more aggressive behavior (tumor budding, perineural invasion, vascular invasion, and infiltrating tumor border) in the surgical specimen. Furthermore, this system identified colorectal cancer with nodal involvement or high pTNM stage with a 78% positive predictive value and 71% and 69% negative predictive values, respectively. Our findings suggest that a grading system based on the quantification of poorly differentiated clusters is feasible in most colorectal cancer endoscopic biopsies. In view of its good reproducibility, accuracy, and predictive value on the anatomical extent of the disease, it may be taken into account for decision-making in colorectal cancer treatment. Â© 2014 Elsevier Inc. All rights reserved.

2014 - Incidence, clinical features and possible etiology of early onset (≤40 years) colorectal neoplasms. [Articolo su rivista]
Domati, Federica; Maffei, Stefania; Kaleci, Shaniko; Di Gregorio, C; Pedroni, Monica; Roncucci, Luca; Benatti, Piero; Magnani, Giulia; Marcheselli, Luigi; Bonetti, Lr; Mariani, Francesco; Alberti, Am; Rossi, V; PONZ DE LEON, Maurizio
abstract

The aim of the study was to investigate the clinical features, including survival, of patients with colorectal malignancies developed at a very early age (≤40 years), together with possible factors involved in the pathogenesis of these rare neoplasms. The study took advantage of the existence of a specialized colorectal cancer Registry active from 1984. 57 patients met the criteria of early onset cancer; main epidemiological data, morphology, stage, familial aggregation, possible role of inheritance and survival were analyzed. Despite the relevant increase over time of all registered patients, joiningpoint analysis of crude incidence rate of early onset colorectal neoplasms revealed a certain stability of these tumors (EAPC: 2.4, CI 14-22) with a constant prevalence of the male sex. Stage at diagnosis did not show significant variations between early onset and maturity onset colorectal neoplasms. Hereditary as well as familial cases were significantly (P < 0.005 and 0.03) more frequent among patients with early onset tumors, although in the majority of them no specific etiological factor could be identified. Survival was more favorable in patients with early onset tumors, though this had to be attributed to the higher presence of some histological types in early onset cases. Survival was significantly more favorable for patients of all ages registered in the last decade. Incidence of early onset colorectal cancer was relatively stable between 1984 and 2008. A male preponderance was evident through the registration period. Hereditary and familial cases were significantly more frequent among early onset case. A well defined etiology could be observed in 16% of the cases (versus 2-3% in older individuals). Five-year survival showed a significant improvement over time.

2014 - Inflammation: an important parameter in the search of prostate cancer biomarkers [Articolo su rivista]
Bergamini, Stefania; Bellei, Elisa; REGGIANI BONETTI, Luca; Monari, Emanuela; Cuoghi, Aurora; Francesco, Borelli; Sighinolfi, Maria Chiara; Bianchi, Giampaolo; T., Ozben; Tomasi, Aldo
abstract

Background A more specific and early diagnostics for prostate cancer (PCa) is highly desirable. In this study, being inflammation the focus of our effort, serum protein profiles were analyzed in order to investigate if this parameter could interfere with the search of discriminating proteins between PCa and benign prostatic hyperplasia (BPH). Methods Patients with clinical suspect of PCa and candidates for trans-rectal ultrasound guided prostate biopsy (TRUS) were enrolled. Histological specimens were examined in order to grade and classify the tumor, identify BPH and detect inflammation. Surface Enhanced Laser Desorption/Ionization-Time of Flight-Mass Spectrometry (SELDI-ToF-MS) and two-dimensional gel electrophoresis (2-DE) coupled with Liquid Chromatography-MS/MS (LC-MS/MS) were used to analyze immuno-depleted serum samples from patients with PCa and BPH. Results The comparison between PCa (with and without inflammation) and BPH (with and without inflammation) serum samples by SELDI-ToF-MS analysis did not show differences in protein expression, while changes were only observed when the concomitant presence of inflammation was taken into consideration. In fact, when samples with histological sign of inflammation were excluded, 20 significantly different protein peaks were detected. Subsequent comparisons (PCa with inflammation vs PCa without inflammation, and BPH with inflammation vs BPH without inflammation) showed that 16 proteins appeared to be modified in the presence of inflammation, while 4 protein peaks were not modified. With 2-DE analysis, comparing PCa without inflammation vs PCa with inflammation, and BPH without inflammation vs the same condition in the presence of inflammation, were identified 29 and 25 differentially expressed protein spots, respectively. Excluding samples with inflammation the comparison between PCa vs BPH showed 9 unique PCa proteins, 4 of which overlapped with those previously identified in the presence of inflammation, while other 2 were new proteins, not identified in our previous comparisons. Conclusions The present study indicates that inflammation might be a confounding parameter during the proteomic research of candidate biomarkers of PCa. These results indicate that some possible biomarker-candidate proteins are strongly influenced by the presence of inflammation, hence only a well-selected protein pattern should be considered for potential marker of PCa.

2014 - Italian cancer figures, report 2014: Prevalence and cure of cancer in Italy [Articolo su rivista]
Adamo, Ms; Alessi, D; Aletta, P; Amodio, R; Andreone, S; Angelin, T; Anghinoni, E; Annulli, Ml; Arciprete, C; Artioli, Me; Autelitano, M; Baili, P; Balducci, C; Baracco, M; Baracco, S; Battisti, W; Bella, F; Bellatalla, C; Bellini, A; Belluardo, C; Benatti, P; Benedetto, G; Benfatto, L; Bernazza, E; Bianconi, F; Biavati, P; Bidoli, E; Birri, S; Bizzoco, S; Bonelli, L; Bonini, A; Borciani, E; Bordini, M; Bovo, E; Bozzani, F; Braghiroli, B; Brucculeri, Ma; Brunori, V; Bucalo, G; Bucchi, L; Bugliarello, E; Bulatko, A; Busco, S; Busso, P; Buzzoni, C; Calabrese, A; Calabretta, L; Caldarella, A; Candela, G; Cannone, G; Canu, L; Caparelli, M; Capocaccia, R; Cappelletti, M; Caprara, L; Carboni, D; Carletti, N; Caroli, S; Cascio, Ma; Cascone, G; Casella, C; Castaing, M; Cavalieri d'Oro, L; Cecconami, L; Celesia, Mv; Cena, T; Cercato, Mc; Cesaraccio, R; Chiesa, R; Cirilli, C; Cocchioni, M; Codazzi, T; Cogno, R; Colamartini, A; Colanino Ziino, A; Cometti, I; Contiero, P; Contrino, Ml; Corbinelli, A; Cordaro, C; Corti, M; Costa, A; Costarelli, D; Coviello, V; Crapanzano, G; Cremone, L; Crocetti, E; Cuccaro, F; Curatella, S; Cusimano, R; D'Alò, D; Dal Cappello, T; Dal Cin, A; Dal Maso, L; Davini, C; De Dottori, M; De Angelis, R; De Santis, E; De Valiere, E; Dei Tos, Ap; Demurtas, G; Devigili, E; Di Felice, E; di Grazia, L; Di Gregorio, C; di Norcia, R; Di Prima, A; Dinaro, Y; Distefano, R; Doa, N; Domati, F; Fabiano, S; Facchinelli, G; Falcini, F; Falk, M; Fanetti, Ac; Fattoruso, S; Federico, Massimo; Ferrari, F; Ferrari, L; Ferretti, S; Fidelbo, M; Filipazzi, L; Fiore, Ar; Fiori, G; Foca, F; Forgiarini, O; Foschi, R; Francisci, S; Frasca, G; Frassoldi, E; Fusco, M; Fusco, M; Gada, D; Garrone, E; Gasparotti, C; Gatta, G; Gatti, L; Gaudiano, C; Gennaro, V; Gentilini, Ma; Gerevini, C; Ghilardi, S; Ghisleni, S; Giacomin, A; Giavazzi, L; Gigli, A; Gilardi, F; Giorgetti, S; Giorgi Rossi, P; Giubelli, C; Giuliani, O; Giurdanella, Mc; Gola, G; Goldoni, Ca; Golizia, Mg; Greco, A; Guarda, L; Guttadauro, A; Guzzinati, S; Iachetta, F; Iannelli, A; Ieni, A; Intrieri, T; Kaleci, S; La Rosa, F; Lando, C; Lavecchia, Am; Lazzarato, F; Le Rose, L; Leone, A; Leone, R; Lonati, F; Lucchi, S; Luminari, Stefano; Macci, L; Macerata, V; Madeddu, A; Maffei, S; Maghini, A; Magnani, C; Magnani, G; Magoni, M; Mallone, S; Mameli, G; Mancini, S; Mancuso, P; Mangone, L; Manneschi, G; Mannino, R; Mannino, S; Marani, E; Marchesi, C; Mariani, F; Martorana, C; Marzola, L; Maspero, S; Maule, M; Mazzei, A; Mazzoleni, G; Mazzucco, G; Melcarne, A; Merletti, F; Merlo, E; Michiara, M; Migliari, E; Minerba, S; Minicuzzi, A; Mizzi, M; Monetti, D; Morana, G; Moroni, E; Mosso, Ml; Muni, A; Mura, F; Natali, M; Negrino, L; Nemcova, L; Nicita, C; Ocello, C; Pala, F; Palumbo, M; Panciroli, E; Panico, M; Pannozzo, F; Pascucci, C; Pasolini, A; Pastore, G; Patriarca, S; Pedroni, M; Perrotta, C; Pesce, P; Petrinelli, Am; Petrucci, C; Pezzarossi, A; Pezzuto, L; Piffer, S; Pinon, M; Pinto, A; Pintori, N; Pirani, M; Pirino, D; Pironi, V; Ponz de Leon, M; Prandi, R; Prazzoli, R; Puleio, M; Puppo, A; Quarta, F; Quattrocchi, M; Ramazzotti, V; Rashid, I; Ravaioli, A; Ravazzolo, B; Ravegnani, M; Reggiani Bonetti, L; Ricci, P; Rinaldi, E; Rizzello, R; Rognoni, M; Rollo, Pc; Roncaglia, F; Roncucci, Luca; Rosano, A; Rossi, F; Rossi, G; Rossi, M; Rossi, S; Rossini, S; Rosso, S; Rudisi, G; Ruggeri, Mg; Russo, Ag; Russo, M; Sacchettini, C; Sacchetto, L; Sacco, G; Sacerdote, C; Salvatore, S; Salvi, O; Sampietro, G; Santucci, C; Scheibel, M; Sciacca, S; Sciacchitano, C; Sciacchitano, S; Scuderi, T; Sechi, O; Seghini, P; Senatore, G; Serafini, G; Serraino, D; Sgargi, P; Sini, Gm; Sobrato, I; Soddu, M; Solimene, C; Spano, F; Spata, E; Sperduti, I; Spinosa, S; Staiti, R; Stocco, C; Stracci, F; Sunseri, R; Sutera Sardo, A; Tagliabue, G; Tamburo, L; Tamburrino, S; Taranto, V; Terracini, B; Tisano, F; Tittarelli, A; Tognazzo, S; Torrisi, A; Torrisi, A; Traina, A;
abstract

This Report intends to estimate the total number of people still alive in 2010 after cancer diagnosis in Italy, regardless of the time since diagnosis, and to project these estimates to 2015. This study is also aimed to estimate the number of already cured cancer patients, whose mortality rates have become undistinguishable from that of the general population of the same age and sex.

2014 - Measurement of Î²-tryptase in postmortem serum in cardiac deaths [Articolo su rivista]
Palmiere, Cristian; Comment, Lionel; Vilarino, Raquel; Mangin, Patrice; Reggiani Bonetti, Luca
abstract

Mast cells are well known for their role in hypersensitivity reactions. However, there is increasing evidence that they might also participate in both developing and weakening atherosclerotic plaques, potentially causing plaque instability. Some clinical studies have therefore postulated the existence of relationships between blood Î²-tryptase levels and acute coronary syndromes. In this study, we investigated postmortem serum Î²-tryptase levels in a series of 90 autopsy cases with various degrees of coronary atherosclerosis that had undergone medico-legal investigations. Î²-tryptase concentrations in these cases were compared to levels observed in 6 fatal anaphylaxis cases following contrast material administration. Postmortem serum Î²-tryptase concentrations in the anaphylactic deaths ranged from 146 to 979 ng/ml. In 9 out of 90 cases of cardiac deaths, Î²-tryptase levels were higher than clinical reference values of 11.4 ng/ml and ranged from 21 to 65 ng/ml. These results indicate that increased postmortem serum Î²-tryptase levels can be observed, though not systematically, in cardiac deaths with varying degrees of coronary atherosclerosis disease, thereby suggesting that mast cell activation in this disease cannot be ascertained by postmortem serum Î²-tryptase measurements. Â© 2014 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

2014 - Measurement of Î²-tryptase in postmortem serum, pericardial fluid, urine and vitreous humor in the forensic setting [Articolo su rivista]
Comment, Lionel; Reggiani Bonetti, Luca; Mangin, Patrice; Palmiere, Cristian
abstract

In the realm of forensic pathology, Î²-tryptase measurement for diagnostic purposes is performed in postmortem serum obtained from femoral blood. This may be partially or completely unavailable in some specific cases, such as infant autopsies and severely damaged bodies. The aim of this study was to investigate the usefulness of determining Î²-tryptase levels for diagnostic purposes in alternative biological samples. Urine, vitreous humor and pericardial fluid were selected and measured in 94 subjects including: fatal anaphylaxis following contrast material administration (6 cases), hypothermia (10 cases), diabetic ketoacidosis (10 cases), gunshot suicide (10 cases), heroin injection-related deaths (18 cases), trauma (10 cases), sudden death with minimal coronary atherosclerosis (10 cases), severe coronary atherosclerosis without myocardial infarction (10 cases) and severe coronary atherosclerosis with myocardial infarction (10 cases). Postmortem serum and pericardial fluid Î²-tryptase levels higher than the clinical reference value (11.4 ng/ml) were systematically identified in fatal anaphylaxis following contrast material administration and 6 cases unrelated to anaphylaxis. Î²-tryptase concentrations in urine and vitreous humor were lower than the clinical reference value in all cases included in this study. Determination of Î²-tryptase in pericardial fluid appears to be a possible alternative to postmortem serum in the early postmortem period when femoral blood cannot be collected during autopsy and biochemical investigations are required to objectify increased Î²-tryptase levels. Â© 2014 Elsevier Ireland Ltd.

2014 - Poorly differentiated clusters (PDCs) as a novel histological predictor of nodal metastases in pT1 colorectal cancer [Articolo su rivista]
Barresi, Valeria; Branca, Giovanni; Ieni, Antonio; Reggiani Bonetti, Luca; Baron, Luigi; Mondello, Stefania; Tuccari, Giovanni
abstract

The practical use of histological factors such as submucosal (SM) invasion depth, poor differentiation, presence of lymphovascular invasion (LVI) and tumour budding to establish the risk of nodal dissemination in pT1 colorectal cancer (CRC) is limited by their low standardization and high inter-observer variability. It was recently suggested that the presence in CRC histological sections of poorly differentiated clusters (PDCs), defined as â¥5 cancer cells with no gland formation, may predict the metastatic potential of CRC. In addition, PDC assessment was shown to be more reproducible than the evaluation of the other aforementioned histological predictors. Hence, in this study, we investigated and compared the predictive value of PDC and other histological parameters on the risk of nodal involvement in pT1 CRC. The presence of PDC, SM invasion depth â¥1,000 Î¼m and LVI was significantly associated with N+ status in pT1 CRC (P<0.0001). Among these parameters, SM invasion depth had the highest sensitivity to identify N+ pT1 CRC but with the lowest specificity. When the analysis was restricted to CRCs with SM invasion depth â¥1,000 Î¼m, the presence of PDC was the only independent risk factor for nodal metastases and allowed the identification of 87.5 % of N+ cancers. In conclusion, in this study, we demonstrate that the presence of PDC is associated with the metastatic potential of pT1 CRC. The combination of this parameter with SM invasion depth may allow identifying most of the pT1 CRC with nodal metastases. Â© 2014 Springer-Verlag.

2014 - Postmortem findings in bone cement implantation syndrome-related deaths [Articolo su rivista]
De Froidmont, Sébastien; Bonetti, Luca Reggiani; Villaverde, Raquel Vilariño; Del Mar Lesta, Maria; Palmiere, Cristian
abstract

Several mechanisms have been postulated as potentially involved in life-threatening complications during cemented surgery. In this study, we evaluated the role of anaphylaxis and pulmonary fat embolism in the pathophysiology of bone cement implantation syndrome in a series of fatal cases that underwent medicolegal investigations. Postmortem findings in these cases were compared with those obtained from individuals who died after other injuries and/or interventions and in which activated mast cells and pulmonary fat embolism were involved in the pathogenesis of death. Fifty subjects were selected including 6 individuals who had undergone cemented total hip arthroplasty and died intraoperatively, 32 subjects who died shortly after being involved in traffic accidents, 8 individuals who died shortly after the injection of contrast material, and 4 subjects who had undergone orthopedic surgery and died postoperatively. Massive pulmonary fat embolism was determined to be the cause of death in all the 6 subjects who died intraoperatively as well as the main cause of death in traffic-road victims with rapid respiratory function deterioration. Mast cell activation was identified exclusively in the group of subjects who died shortly after contrast material administration. Massive pulmonary fat embolism appears to be the most important factor responsible for severe cardiorespiratory function deterioration during cemented arthroplasty. Cardiac comorbidities can also significantly influence the severity of intraoperative complications, thus corroborating the hypothesis of a multifactorial model in the pathogenesis of bone cement implantation syndrome. Copyright Â© 2014 by Lippincott Williams & Wilkins.

2014 - Risk factors in fatal cases of anaphylaxis due to contrast media: A forensic evaluation [Articolo su rivista]
Palmiere, Cristian; Bonetti, Luca Reggiani
abstract

Background: Fatalities following contrast medium exposure are extremely rare in clinical routine, though they may occur as an exception. Some may fall under the purview of the inquiring authorities and forensic pathologists due to their inhospital occurrence. The purpose of this study was to assess the risk factors for anaphylaxis due to contrast medium administration that can be identified in fatal cases.Methods: Fatalities occurred during the course of clinical investigations with contrast media described in the literature and fatal reactions to contrast agents that had undergone forensic investigations in our medicolegal center were reviewed with respect to patient characteristics, administered contrast medium, performed biochemical investigations and potential risk factors identified based on clinical history and medical records.Results: Biochemical investigations into the fatal cases examined in our facility revealed increased mast cell tryptase, total IgE and activated mast cells in all subjects. Data obtained from the literature and our own investigations indicated that in only a minority of the fatal cases had there been previous exposure to contrast compounds, while most cases of severe anaphylaxis involved patients who apparently reacted on initial exposure.Conclusions: Most fatal cases failed to present any identifiable predisposing conditions out of those traditionally considered as risk factors for an anaphylactic reaction to contrast compounds in the medical histories of the patients. Comprehensive clinical histories and thorough reviews of medical data, along with exhaustive forensic investigations, provide information that is relevant in order to better appreciate the interwoven relationships linking all factors potentially involved in the pathogenesis of fatal anaphylaxis to contrast media.

2014 - Splenic hypereosinophilia in anaphylaxis-related death: different assessments depending on different types of allergens? [Articolo su rivista]
Reggiani Bonetti, Luca; Maccio, Livia; Trani, Nicoletta; Radheshi, Erjon; Palmiere, Cristian
abstract

The aim of this study was to evaluate splenic eosinophil and mast cell accumulation using pagoda red stain in a series of anaphylaxis-related deaths that underwent medico-legal investigations. Our goal was to assess whether fatal reactions to insect stings, intramuscularly administered antibiotics and intravenously injected contrast media are responsible for specific patterns of eosinophil and mast cell accumulation. Two study groups were prospectively formed, an anaphylaxis-related death group and a control group. Autopsy, histology (haematoxylin-eosin stain, pagoda red stain and immunohistochemistry using anti-tryptase antibodies), toxicology and postmortem biochemistry (beta-tryptase, total IgE and specific IgE) were performed in all cases. All tested parameters (spleen weight, beta-tryptase and total IgE levels as well as eosinophil, mast cell and degranulated mast cell numbers in the spleen) were significantly higher in the anaphylaxis-related death group. No statistically significant differences were observed among the various groups (intramuscular antibiotic injection, intravenous contrast medium administration and stinging insects) in any combination, suggesting that mast cell and eosinophil accumulation in the spleen during anaphylaxis does not have any specific pattern related to the triggering allergen. Despite a lower sensitivity than immunohistochemical staining in discriminating eosinophil and mast cells, pagoda red stain allowed these cells to be identified and could therefore be proposed as a low-cost, first-line diagnostic procedure in those situations where immunohistochemistry is not systematically performed or cannot be carried out.

2014 - Validation of prostate cancer biomarkers and inflammation: a proteomic study [Abstract in Atti di Convegno]
Bergamini, S; Bellei, E; Monari, E; Cuoghi, A; Reggiani Bonetti, L; Borelli, F; Sighinolfi M., C; Bianchi, G; Ozben, T; Tomasi, A
abstract

Validation of prostate cancer biomarkers and inflammation by a proteomic approach

2013 - Evaluation of the effects of cryopreservation on Modena Biobank tissue samples through HR-MAS NMR [Abstract in Atti di Convegno]
Valeria, Righi; Schenetti, Luisa; Maiorana, Antonino; Bettelli, Stefania Raffaella; REGGIANI BONETTI, Luca; Mucci, Adele
abstract

The application of high-resolution magic angle spinning (HR-MAS) NMR spectroscopy to the analysis of intact tissue biopsies and surgery samples dates back to 1997. It represents, at present, an actively investigated field, and it is used to obtain the metabolic fingerprints of tissues. It has been shown to differentiate between normal and neoplastic tissue in the case of breast, brain, kidney, colon cancer and other malignancies in the upper gastrointestinal tract. Nevertheless, it is difficult to run HR-MAS analysis immediately after surgical resection or biopsy, and samples are currently frozen in liquid nitrogen and then stored to -80 °C. Some studies reporting the effect of sample ischaemia, spinning time and measurement temperature on the metabolic profile of tissues can be found.1 Nevertheless, another important issue to be addressed is the effect of cryopreservation time on the metabolic profile of tissues. Crypreservation represents a more and more common practice, due to the widespread of tissue BioBanks. To gain a deeper insight into metabolic changes that occur during the storage, samples of Modena BioBank were analyzed through HR-MAS NMR after 1, 6 and 12 months from collection. The results of this study were discussed.

2013 - Incidence trend of malignant polyps through the data of a specialized colorectal cancer registry: clinical features and effect of screening. [Articolo su rivista]
REGGIANI BONETTI, Luca; DI GREGORIO, Carmela; Pedroni, Monica; Domati, Federica; Barresi, V; Marcheselli, L; PONZ DE LEON, Maurizio
abstract

OBJECTIVES: The purposes of the study are to describe the incidence trend of malignant polyp of large bowel over a 25-year period in the District of Modena and to assess the effect of an organized colorectal cancer screening program. MATERIAL AND METHODS: Through the data of a specialized colorectal cancer Registry, we evaluate the clinical and pathological features of the polyps. Trend analysis was assessed with the Joinpoint Regression Program. RESULTS: A total of 172 patients with malignant polyps were diagnosed throughout the study (3.5% of 4.835 registered patients); their overall frequency during the registration period increased from zero cases in the initial years (1984-85) to 57 cases in the past 3 years (2006-2008). Crude incidence rate passed from 0.37 in 1986-89 to 10.2 in 2006. Joinpoint trend analysis of crude rates showed a significant increase of incidence during the study period, with percent of annual variation ranging between 38.6% (95% CI 12.5-70.7) and 7.3% (95% CI 2.6-12.1). During the screening period (2005-2008, the past 4 years of registration) there was a significant increase of sessile polyps (p < 0.001), while other clinical and morphological features, including the number of low- and high-risk malignant polyps, remained unchanged. The surgery (after polypectomy) tended to raise both in low- and high-risk subgroups. CONCLUSION: The incidence of malignant polyps increased significantly from the initial to the most recent periods of colorectal cancer registration. Screening was associated with changes in gross morphology of polyps and with an increased use of the surgery after endoscopic polypectomy.

2013 - Italian cancer figures, report 2013: Multiple tumours [Articolo su rivista]
Adamo, Ms; Alessi, D; Aletta, P; Amodio, R; Andreone, S; Angelin, T; Anghinoni, E; Annulli, Ml; Antonini, S; Artioli, Me; Autelitano, M; Balducci, C; Balottari, P; Baracco, M; Battisti, W; Bella, F; Bellatalla, C; Belluardo, C; Benatti, Piero; Benedetto, G; Benfatto, L; Bernazza, E; Bianconi, F; Biavati, P; Bidoli, E; Birri, S; Bizzoco, S; Bonelli, L; Bonini, A; Borciani, E; Bovo, E; Bozzani, F; Bozzeda, A; Braghiroli, B; Brucculeri, Ma; Brunori, V; Bucalo, G; Bucchi, L; Bugliarello, E; Bulatko, A; Busco, S; Busso, P; Buzzoni, C; Calabretta, L; Caldarella, A; Candela, G; Canu, L; Cappelletti, M; Caprara, L; Carboni, D; Carletti, N; Caroli, S; Carone, S; Cascio, Ma; Cascone, G; Casella, C; Castaing, M; Cecconami, L; Celesia, Mv; Cena, T; Cercato, Mc; Cesaraccio, R; Chiesa, R; Cirilli, C; Civaschi, A; Cocchioni, M; Codazzi, T; Cogno, R; Colamartini, A; Colanino Ziino, A; Cometti, I; Contiero, P; Contrino, Ml; Corbinelli, A; Cordaro, C; Corti, M; Costa, A; Costarelli, D; Cremone, L; Crocetti, E; Curatella, S; Cusimano, R; D'Alò, D; D'Angelo, S; Dal Cappello, T; Dal Cin, A; Dal Maso, L; Dall'Acqua, M; Dalsasso, F; Davini, C; De Dottori, M; De Maria, V; De Santis, E; De Valiere, E; Dei Tos, Ap; Demurtas, G; Devigli, E; Di Felice, E; di Grazia, L; Di Gregorio, C; Di Prima, A; Distefano, R; Doa, N; Domati, F; Fabiano, S; Facchinelli, G; Falcini, F; Falk, M; Fanetti, Ac; Fattoruso, S; Federico, Massimo; Ferrari, L; Ferretti, S; Fidelbo, M; Filipazzi, L; Fiore, Ar; Fiori, G; Foca, F; Forgiarini, O; Frasca, G; Frassoldi, E; Frizza, J; Fusco, M; Fusco, M; Gada, D; Garrone, E; Gasparotti, C; Gatti, L; Gaudiano, C; Gennaro, V; Gentilini, M; Gerevini, C; Ghilardi, S; Ghisleni, S; Giacomin, A; Giavazzi, L; Gilardi, F; Giorgetti, S; Giubelli, C; Giuliani, O; Giurdanella, Mc; Gola, G; Goldoni, Ca; Golizia, Mg; Grandi, L; Greco, A; Guarda, L; Guttadauro, A; Guzzinati, S; Iachetta, F; Iannelli, A; Ieni, A; Intrieri, T; Kaleci, S; La Rosa, F; Lando, C; Lavecchia, Am; Lazzarato, F; Leone, A; Leone, R; Lonati, F; Lottero, B; Lucchi, S; Luminari, Stefano; Macci, L; Macerata, V; Madeddu, A; Maffei, S; Maghini, A; Magnani, C; Magnani, G; Magoni, M; Mameli, G; Mancini, S; Mancuso, P; Mangone, L; Manneschi, G; Mannino, R; Mannino, S; Marani, E; Mariani, F; Martorana, C; Marzola, L; Maspero, S; Maule, M; Mazzei, A; Mazzoleni, G; Mazzucco, G; Melcarne, A; Merletti, F; Michiara, M; Migliari, E; Minerba, S; Minicuzzi, A; Mizzi, M; Monetti, D; Morana, G; Moroni, E; Mosso, Ml; Muni, A; Mura, F; Natali, M; Nemcova, L; Nicita, C; Ocello, C; Paci, E; Pala, F; Palumbo, M; Panico, M; Pannozzo, F; Pascucci, C; Pastore, G; Patriarca, S; Pedroni, Monica; Pellegri, C; Perrotta, C; Pesce, P; Petrinelli, Am; Petrucci, C; Pezzarossi, A; Piffer, S; Pintori, N; Pirani, M; Pirino, D; Pironi, V; PONZ DE LEON, Maurizio; Prandi, R; Prazzoli, R; Preto, L; Puleio, M; Puppo, A; Quaglia, A; Quarta, F; Quattrocchi, M; Raho, Am; Ramazzotti, V; Rashid, I; Ravaioli, A; Ravazzolo, B; Ravegnani, M; Reggiani Bonetti, L; Ribaudo, M; Rinaldi, E; Ricci, P; Rizzello, R; Rollo, Pc; Roncucci, Luca; Rosano, A; Rossi, F; Rossi, G; Rossi, M; Rossini, S; Rosso, S; Rudisi, G; Ruggeri, Mg; Russo, Ag; Russo, M; Sacchettini, C; Sacco, G; Sacerdote, C; Salvatore, S; Salvi, O; Sampietro, G; Sandrini, M; Santucci, C; Scheibel, M; Schiacchitano, S; Sciacca, S; Sciacchitano, C; Scuderi, T; Sechi, O; Seghini, P; Senatore, G; Serafini, G; Serraino, D; Sgargi, P; Sigona, A; Sini, Gm; Sobrato, I; Soddu, M; Solimene, C; Spano, F; Spata, E; Sperduti, I; Staiti, R; Stocco, C; Stracci, F; Sunseri, R; Sardo, As; Tagliabue, G; Tamburo, L; Tamburrino, S; Tanzarella, M; Terracini, B; Tessandori, R; Tisano, F; Tittarelli, A; Tognazzo, S; Torrisi, A; Torrisi, A; Traina, A; Trapani, C; Tschugguel, B; Tumino, R; Usala, M; Vacirca, S; Valerio, O; Valla, K; Varvarà, M; Vasquez, E; Vassante, B; Vattiato, R; Vercelli, M; Vercellino, Pc; Vicentini, M; Villa, M; Vitale, F; Vital
abstract

OBJECTIVES: This collaborative study, based on data collected by the network of Italian association of cancer registries (AIRTUM), provides updated estimates on the incidence risk of multiple primary cancer (MP). The objective is to highlight and quantify the bidirectional associations between different oncological diseases. The quantification of the excess or decreased risk of further cancers in cancer patients, in comparison with the general population, may contribute to understand the aetiology of cancer and to address clinical follow-up. MATERIAL AND METHODS: Data herein presented were provided by AIRTUM population-based cancer registries, which cover nowadays 48% of the Italian population. This monograph utilizes the AIRTUM database (December 2012), considering all malignant cancer cases diagnosed between 1976 and 2010. All cases are coded according to ICD-O-3. Non-melanoma skin cancer cases, cases based on death certificate only, cases based on autopsy only, and cases with follow-up time equal to zero were excluded. To define multiple primaries, IARC-IACR rules were adopted (http://www.iacr.com.fr/MPrules_july2004.pdf). Data were subjected to standard quality control procedures (described in the AIRTUM data management protocol) and specific quality control checks defined for the present study. A cohort of cancer patients was followed over time from first cancer diagnosis until the date of second cancer diagnosis, death, or the end of follow-up, to evaluate whether the number of observed second cancer cases was greater than expected. Person years at risk (PY) were computed by first cancer site, geographic area (North, Centre, South and Islands), attained age, and attained calendar-year group. All second cancers diagnosed in the cohort's patients were included in the observed numbers of cases. The expected number of cancer cases was computed multiplying the accumulated PY by the expected rates, calculated from the AIRTUM database stratified by cancer site, geographic area, age, and calendar-year group. The Standardized Incidence Ratio (SIR) was calculated as the ratio of observed to expected cancer cases. The Excess Absolute Risk (EAR) beyond the expected amount were calculated subtracting the expected number of subsequent cancers from the observed number of cancer cases; the difference was then divided by the PY and the number of cancer cases in excess (or deficit) was expressed per 1,000 PY. Confidence intervals were stated at 95%. The two months (60 days) after first cancer diagnosis were defined as "synchronicity period", and in the main analysis observed and expected cases during this period were excluded. It was estimated the excess risk in the period after first diagnosis (≥ 0 months), excluding the synchronicity period (≥ 2 months), and during the following periods: 2-11, 12-59, 60-119 and 120 months after diagnosis. First-cancer-site-and-gender-specific sheets are presented, reporting both SIRs and EARs. RESULTS: For 5,979,338 person-years a cohort of 1,635,060 cancer patients (880,361 males and 754,699 females) diagnosed between 1976 and 2010 was followed. The mean follow-up length was 14 years. Overall, 85,399 metachronous (latency ≥2 months) cancers were observed, while 77,813 were expected during the study period: SIR: 1.10 (95%CI 1.09-1.10), EAR: 1.32 x 1,000 person-years (95%CI 1.19 - 1.46). The SIR was 1.08 (95%CI 1.08-1.09) for men (54,518 observed and 50,260 expected) and 1.12 (95%CI 1.11-1.13) for women (30,881/27,553), and the EAR 1.61 (95%CI 1.37-1.84) and 1.08 x 1,000 person-years (95%CI 0.93-1.24), respectively.Moreover, during the first two months after first cancer diagnosis (synchronous period) 14,807 cancers were observed while 3,536 were expected (SIR: 4.16; 95%CI 4.09-4.22); the SIR was 4.08 (95%CI 4.00-4.16) for men and 4.32 (95%CI 4.20-4.45) for women.The mean age of patients at first cancer diagnosis was 67.0 years among males and

2013 - Lack of evidence for an association between seminoma and human papillomavirus infection using GP5+/GP6+ consensus primers. [Articolo su rivista]
Bertazzoni, G; Sgambato, A; Migaldi, Mario; Grottola, Antonella; Sabbatini, Am; Nanni, N; Farinetti, Alberto; Iachetta, F; Giacobazzi, E; Pecorari, M; Bonetti, L. R.
abstract

Testicular germ cell tumors account for about 1% of all cancers. The incidence of these tumors is increasing and they represent the most common solid malignancies of young men aged 15– 40 years with seminoma being one of the most common histotype. Pathogenesis of testicular germ cell tumors remains unknown and, although cryptorchidism is considered the main risk factor, there is evidence of an association with environmental and genetic risk factors. Human papillomaviruses (HPV) are a family of DNA viruses and represent a major risk factor for cervical cancer. In addition, they have been associated with other human non-malignant and malignant diseases, including breast and head and neck cancer. HPV sequences have been detected throughout the male lower genitourinary tract as well as in seminal fluid and an increased testicular tumorigenesis has been reported in HPV transgenic mice. Aim of this study was to evaluate the potential involvement of HPV in human testicular tumorigenesis. Realtime PCR employing GP5þ/GP6þ consensus HPV primers was used to examine the presence of HPV sequences in a subset of human seminoma (n ¼ 61) and normal testicles (n ¼ 23). None of the specimens tested displayed the presence of HPV DNA. These findings do not support an association between HPV and human seminoma and warrant further studies to assess definitively the role of these viruses in human testicular tumorigenesis.

2013 - Lymph node evaluation in stage IIA colorectal cancer and its impact on patient prognosis: A population-based study [Articolo su rivista]
Iachetta, Francesco; REGGIANI BONETTI, Luca; Marcheselli, Luigi; DI GREGORIO, Carmela; Cirilli, C.; Messinese, S.; Cervo, Gian Luca; Postiglione, Raffaella; DI EMIDIO, Katia; Pedroni, Monica; Longinotti, E.; Federico, Massimo; PONZ DE LEON, Maurizio
abstract

Background. The analysis of regional lymph nodes is particularly relevant in patients with stage II colorectal cancer, in whom the role of adjuvant chemotherapy remains unclear. The aim of this study was to assess the relationship between number of examined lymph nodes and survival in patients with stage IIA (pT3N0M0) colorectal cancer, and to determine the optimal number of lymph nodes that should be examined. Methods. The study group included all the surgically-treated colorectal cancer patients in stage IIA (n = 657) who were identified through the population-based Cancer Registry of the Province of Modena (Northern Italy), during the period 2002-2006. Results. The median number of harvested lymph nodes was 19 (range 1-68). Considering, as a reference point, patients with 12 or less lymph nodes, subjects with n ≥ 20 lymph nodes examined showed, in univariate analysis, a significantly higher cancer specific (p = 0.01) and relapse-free survival (p = 0.003). The results were confirmed by multivariate analysis (Cox model). Conclusion. The result suggests that colorectal cancer patients in stage IIA with n ≥ 20 lymph nodes examined exhibit better survival when compared with subjects in whom fewer lymph nodes were examined. The number of 20 lymph nodes is the essential requirement for an oncologic resection of the large bowel.

2013 - Sodium/iodide symporter is expressed in the majority of seminomas and embryonal testicular carcinomas [Articolo su rivista]
Micali, Salvatore; Maggisano, V.; Cesinaro, A.; Celano, M.; Territo, A.; Reggiani Bonetti, L.; Sponziello, M.; Migaldi, Mario; Navarra, M.; Bianchi, Giampaolo; Filetti, S.; Russo, D.
abstract

Testicular cancer is the most frequent cancer in young men. The large majority of patients has a good prognosis, but in a small group of tumours the current treatments are not effective. Radioiodine is routinely used in the treatment of thyroid cancer and is currently investigated as a potential therapeutic tool even for extra-thyroid tumours able to concentrate this radioisotope. Expression of Na+/I- symporter (NIS), the glycoprotein responsible for iodide transport, has been demonstrated in normal testicular tissue. In this study, we analyzed NIS expression in a large series of testicular carcinomas. Our retrospective series included 107 patients operated for testicular tumours: 98 typical seminomas, 6 embryonal carcinomas, 1 mixed embryonal-choriocarcinoma and 2 Leydig cells tumours. Expression and regulation of NIS mRNA and protein levels were also investigated in human embryonal testicular carcinoma cells (NTERA) by real time RT-PCR and western blotting respectively. Immunohistochemical analysis showed presence of NIS in the large majority of seminomas (90/98) and embryonal carcinomas (5/7) of the testis, but not in Leydig cell carcinomas. Expression of NIS protein was significantly associated to the lymphovascular invasion. In NTERA cells treated with the histone deacetylase inhibitors SAHA and valproic acid, a significant increase of NIS mRNA (about 60 and 30 fold vs control, p<0.001 and p<0.01 respectively) and protein levels, resulting in enhanced ability to uptake radioiodine, was observed. Finally, NIS expression in testicular tumours with the more aggressive behavior is of interest for the potential use of targeting NIS to deliver radioiodine in malignant cells.

2013 - The influence of inflammation in the search of discriminatory biomarkers for prostate cancer: a proteomic study [Abstract in Rivista]
Bergamini, Stefania; REGGIANI BONETTI, Luca; Monari, Emanuela; Bellei, Elisa; Cuoghi, Aurora; Majorana, Antonino; Ozben, Tomis; Micali, Salvatore; Sighinolfi, Maria Chiara; Tomasi, Aldo; Bianchi, Giampaolo
abstract

background: Despite the improvements in clinical and surgical practice, prostate cancer (PCa) remains one of the most widespread cancer in male. The serum marker currently used for the diagnosis of PCa is the prostate-specific antigen (PSA), but its increase does not discriminate benign prostatic hyperplasia (BPH) from PCa. In our study, we investigated the serum protein expression of BPH compared to PCa, in order to identify by Surface Enhanced Laser Desorption/Ionization - Time of Flight - Mass Spectrometry (SELDI-ToF-MS) analysis distinctive protein profiles able to unquestionably discriminate patients with a benign prostate condition from those with a malignant situation. Moreover, we considered these conditions focusing on the co-existence of inflammation. Methods: Patients with clinical suspect of PCa (PSA elevation and/or palpable mass at digital rectal exploration) and candidates for trans-rectal ultrasound guided prostate biopsy were enrolled. The analysis of protein profile of 30 patients with PCa and 30 subjects with BPH was carried out. All histological specimens were examined in order to graduate and classify the tumor and to recognize the BPH condition and presence of inflammation, that was classed in chronic and acute and then graduated in mild, moderate and severe. Serum was depleted of the 6 high-abundance proteins by immunoaffinity chromatography prior to SELDI-ToF-MS analysis. Results: The comparison between protein spectra from PCa and BPH considering the inflammation parameter and excluding samples with moderate and/or severe inflammation, identified 17 differentially expressed protein peaks using H50 ProteinChip Array.The analysis of protein profile in presence of inflammation showed different protein peaks in the two groups, some of which overlapped with those found also in the comparison between PCa and BPH in absence of inflammation. Conclusions: The inflammation seems to lead a crucial contribution in the protein profile assessments of these conditions. On the basis of our results, we believe that certain different protein peaks could be reasonably associated to inflammation rather than to cancer. Therefore, inflammation might be a confounding parameter in the search of specific biomarkers to discriminate PCa from BPH.

2012 - Colorectal carcinoma grading by quantifying poorly differentiated cell clusters is more reproducible and provides more robust prognostic information than conventional grading. [Articolo su rivista]
Barresi, V; Reggiani Bonetti, L; Branca, G; Di Gregorio, C; PONZ DE LEON, Maurizio; Tuccari, G.
abstract

The most widely used system to define the histological grade of colorectal carcinoma (CRC) is based on the degree of gland formation. This system suffers from significant interobserver variability which may limit its prognostic value and consequently better standardized criteria for the assessment of histological grading of CRC are needed. The present study aims to evaluate and to compare, in a cohort of postsurgical pTNM stage I CRC, conventional histological grading, and a novel grading system based on the number of poorly differentiated clusters of neoplastic cells, in terms of interobserver reproducibility, prognostic significance on progression-free survival, and association with other clinicopathological characteristics. Grading with both systems was performed by two pathologists independently and blinded to the clinicopathological data. Interobserver agreement was higher when grade was assessed by counting poorly differentiated clusters than by assessing the relative proportion of the glandular component. Contrary to conventional grading, the novel system provided significant prognostic information in terms of progression-free survival and was significantly associated with budding, invasive growth, lymphatic invasion, and occult nodal metastases of CRC. In conclusion, our findings suggest that a tumor grading system based on the number of poorly differentiated clusters is more reproducible and provides better prognostic stratification of pTNM stage I CRC patients than conventional grading.

2012 - Diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) immunoexpression in follicular-patterned lesions of the thyroid gland [Articolo su rivista]
Barresi, Valeria; Vitarelli, Enrica; Bonetti, Luca Reggiani; Tuccari, Giovanni; Barresi, Gaetano
abstract

Neutrophil gelatinase-associated lipocalin (NGAL) is a protein which participates in iron trafficking and which is involved in cancerogenesis and cancer progression. Since its over-expression has been documented in thyroid malignancies in comparison to thyroid normal gland, in the present study, we aimed to determine whether the evaluation of NGAL immunoexpression may be of help in the differential diagnosis of follicular-patterned thyroid lesions. Our additional aim was to test the possible interference of endogenous biotin on the immunohistochemical findings. Thus, all the immunohistochemical procedures, carried out with labeled streptavidin biotin method, were doubly performed, with or without the preliminary inhibition of endogenous biotin. No NGAL staining was found in the normal thyroid gland nor in the nodular colloid goiters or in Hashimoto's thyroiditis. NGAL expression appeared to be significantly more frequent in the malignant tumors in comparison to benign ones (P<0.000001). Even more, NGAL expression appeared to be specific (specificity 93%) for carcinoma and represented a sensitive method (sensitivity 84%), with high negative (88%) and positive (94%) predictive values, as well as high diagnostic accuracy (88%), in the identification of follicular-patterned thyroid malignant tumors. The specificity, positive predictive value and diagnostic accuracy lowered when biotin was not preliminary inhibited, due to the presence of false positives among benign HÃ¼rthle cell tumors. In conclusion, the immunohistochemical detection of NGAL may be helpful in the differential diagnosis between malignant and benign follicular-patterned lesions of the thyroid. The use of biotin free system or the preliminary biotin inhibition is warranted for the detection of NGAL in thyroid samples, especially when dealing with HÃ¼rthle cell tumors. Â© 2012 Springer-Verlag.

2012 - Immunohistochemical assessment of lymphovascular invasion in stage I colorectal carcinoma: prognostic relevance and correlation with nodal micrometastases. [Articolo su rivista]
Barresi, V; REGGIANI BONETTI, Luca; Vitarelli, E; Di Gregorio, C; PONZ DE LEON, Maurizio; Barresi, G.
abstract

Several studies have suggested that the presence of occult nodal metastases (micrometastases) is related to adverse clinical course in stage I colorectal carcinoma. Herein we analyzed the correlation between nodal micrometastases and lymphovascular invasion (LVI) or lymphatic vessel density (LVD) in a series of stage I colorectal carcinomas; the cohort included cases characterized or not characterized by disease progression during the follow-up. In these cases, LVI and LVD were evidenced through the immunohistochemical detection of the specific marker for lymphatic vessels, D2-40. LVI was significantly more frequent in colorectal carcinomas characterized by the presence of micrometastases (P<0.0001), high peritumoral LVD (P<0.0001), and disease progression (P<0.0001). The analysis for progression risk indicated that nodal micrometastases and LVI were significant, negative, independent prognostic parameters associated with shorter disease-free survival of stage I colorectal cancer (P=0.0001; P=0.0242). In conclusion, in this study we demonstrated for the first time that LVI is significantly associated with nodal occult metastases in stage I colorectal carcinoma. In the light of its significant, independent, prognostic value in this neoplasia, the detection of LVI may represent a faster and cheaper tool compared with the time-consuming evaluation of micrometastases to select high-risk patients who may benefit from adjuvant systemic treatment. Furthermore, the assessment of LVI may be applied to establish the likelihood of nodal involvement from carcinomas treated with conservative local excision techniques, which provide no regional nodes for histologic examination.

2012 - Increased expression of CD133 and reduced dystroglycan expression are strong predictors of poor outcome in colon cancer patients. [Articolo su rivista]
Coco, C; Zannoni, Gf; Caredda, E; Sioletic, S; Boninsegna, A; Migaldi, Mario; Rizzo, G; Bonetti, Lr; Genovese, G; Stigliano, E; Cittadini, A; Sgambato, A.
abstract

Background Expression levels of CD133, a cancer stem cell marker, and of the alpha-subunit of the dystroglycan (alpha-DG) complex, have been previously reported to be altered in colorectal cancers. Methods Expression levels of CD133 and alpha-DG were assessed by immunohistochemistry in a series of colon cancers and their prognostic significance was evaluated. Results Scattered cells positive for CD133 were rarely detected at the bases of the crypts in normal colonic mucosa while in cancer cells the median percentage of positive cells was 5% (range 0--80). A significant correlation was observed with pT parameter and tumor stage but not with tumor grade and N status. Recurrence and death from disease were significantly more frequent in CD133-high expressing tumors and Kaplan-Meier curves showed a significant separation between high vs low expressor groups for both disease-free (p = 0.002) and overall (p = 0.008) survival. Expression of alpha-DG was reduced in a significant fraction of tumors but low alpha-DG staining did not correlated with any of the classical clinical-pathological parameters. Recurrence and death from the disease were significantly more frequent in alpha-DG-low expressing tumors and Kaplan-Meier curves showed a significant separation between for both disease-free (p = 0.02) and overall (p = 0.02) survival. Increased expression of CD133, but not loss of alpha-DG, confirmed to be an independent prognostic parameters at a multivariate analysis associated with an increased risk of recurrence (RR = 2.4; p = 0.002) and death (RR = 2.3; p = 0.003). Conclusions Loss of alpha-DG and increased CD133 expression are frequent events in human colon cancer and evaluation of CD133 expression could help to identify high-risk colon cancer patients.

2012 - Increased expression of CD133 is a strong predictor of poor outcome in stage I colorectal cancer patients [Articolo su rivista]
REGGIANI BONETTI, Luca; Migaldi, Mario; Caredda, E; Boninsegna, A; PONZ DE LEON, Maurizio; Di Gregorio, C; Barresi, V; Scannone, D; Danese, S; Cittadini, A; Sgambato, A.
abstract

Objective. Stage I colorectal carcinomas display a highly variable behavior which is not accurately predicted by the available prognostic markers. CD133 is considered a useful marker to identify the so-called cancer stem cells in colorectal cancers (CRCs) and its expression has been shown to have prognostic significance in CRC patients. This study aimed to verify whether immunohistochemical evaluation of CD133 might correlate with the progression risk of stage I CRC patients. Material and methods. Expression levels of the CD133 molecule were analyzed and compared in two series of stage I surgically resected CRC patients showing disease progression and death for the disease and patients with no evidence of disease progression after at least 6 years after surgery. Results. A positive staining for CD133 was detected in 52% of the cases with poor prognosis and only in 9% of the group with good prognosis, and this difference was highly significant (p < 0.001). A significant correlation was detected between CD133 expression and histological parameters, such as tumor budding, vascular invasion, and presence of lymph node micrometastases but not tumor grading, gender, and age. Disease-free survival and cancer-specific survival of CD133 negative tumors were significantly longer compared to positive cases. In multivariate analyses, CD133 staining confirmed to be a predictor of shorter survival independent from vascular invasion but not from lymph nodes micrometastases. Conclusions. These findings demonstrate that CD133 immunostaining is a useful predictor of high risk progression in stage I CRC patients and might help to identify patients eligible for adjuvant chemotherapy.

2012 - Well-differentiated papillary mesothelioma of the epididymis in a man with recurrent haematospermia. [Articolo su rivista]
Bonetti, L. R.; Schirosi, Laura; Sartori, Giuliana; M., Lupi; Maiorana, Antonino
abstract

We present a case of well-differentiated papillary mesothelioma of the epididymis occurring in a 60-year-old man who came to urologic consult after recurrent episodes of haematospermia. The patient denied pain, fever and trauma in genitals. Local examination revealed indolent swelling at the right testicle and ecography localised a well-circumscribed nodule at the epididymis tail, measuring 2 cm in greater diameter, with associated haemorrhagic hydrocele. A nodulectomy was performed and the patient is alive with no evidence of disease 17 months following surgery.

2011 - An unusual case of signet ring cell adenocarcinoma of the prostate [Articolo su rivista]
REGGIANI BONETTI, Luca; Lupi, Massimo; Stauder, E.; Bergamini, Stefania; Scuri, M.; Maiorana, Antonino
abstract

We report the case of a 70-year-old man with symptoms of urinary obstruction and haematuria, with histological diagnosis of primary signet-ring cell carcinoma of the prostate. Almost 90% of the tumour cells contained characteristic intracytoplasmic vacuoles that positively stained with diastase-digested PAS, Alcian blue and mucicarmine. The positive immunostaining for PSA and PSAP confirmed the prostatic origin of the tumour. Although the patient received hormonal therapy, the disease progressed and the patient died 11 months after surgery.

2011 - Can prostatitis to be a confounding parameter in prostatic proteomic profile designation? [Abstract in Rivista]
Bergamini, Stefania; REGGIANI BONETTI, Luca; Monari, Emanuela; Bellei, Elisa; Maiorana, Antonino; Ozben, T.; Tomasi, Aldo; Micali, Salvatore; Bianchi, Giampaolo
abstract

N/A

2011 - Correlation between CD133 expression and mgmt status in recurrences melanoma [Abstract in Rivista]
Migaldi, Mario; Reggiani Bonetti, L.; Cesinaro, A. M.; Maiorana, Antonino; Farinetti, Alberto; Bettelli, S.; Sgambato, A.
abstract

...

2011 - Cytomegalovirus infection of the upper gastrointestinal tract: a clinical and pathological study of 30 cases. [Articolo su rivista]
Reggiani Bonetti, L.; Losi, L.; Di Gregorio, C.; Bertani, Angela; Merighi, A.; Bettelli, S.; Scuri, M.; Maiorana, Antonino
abstract

OBJECTIVE:The study reviews the endoscopic and histological features of human cytomegalovirus (HCMV) infections of the upper gastrointestinal (UGI) tract.MATERIALS AND METHODS:Clinical histories, endoscopic findings and bioptic specimens of 30 cases of HCMV infection of the UGI tract, diagnosed in a University Hospital in a 10-year period, were reviewed. In all cases, viral inclusion bodies were detected in routine histopathological sections and the diagnosis was confirmed with immunohistochemistry.RESULTS:Six patients were HIV+, whereas four had received organ transplantations, one was affected by common variable immunodeficiency and four had a recent history of malignancy. No other pathologic condition was evidenced in the remaining 15 cases. Mucosal alterations were endoscopically observed in the stomach (19 cases), esophagus (9), cardias (6) and duodenum (1), and multiple organs being synchronously affected in five patients (3 HIV+, 2 with history of malignancy). The antropyloric area was the most frequently affected site. Single ulcers were detected in 11 cases and multiple ulcers in 8, whereas mucosal thickenings (in the form of localized thickenings, polyps or rugal hypertrophy) were present in 13 patients. Thickenings of the mucosa were detected only in the stomach. At histology, necrotic material and granulation tissue were associated with moderate or marked lympho-plasmacytic infiltrate and foveolar hyperplasia in ulcerative lesions, whereas lesions labeled as mucosal thickenings showed mild or moderate chronic inflammatory infiltrate and foveolar hyperplasia.CONCLUSIONS:Endoscopic manifestations of UGI tract involvement in HCMV infection are not specific, varying from erythematous mucosa to ulcers to mucosal thickenings.

2011 - Expression of CD133 protein correlates with lymhp node involvement and recurrence in oral squamous cell carcinoma patients [Abstract in Rivista]
Sgambato, A.; Reggiani Bonetti, L.; Maiorana, Antonino; Farinetti, Alberto; Lupi, M.; Migaldi, Mario
abstract

...

2011 - Human papillomavirus and seminoma [Abstract in Rivista]
Reggiani Bonetti, L.; Migaldi, Mario; Pecorari, M.; Bertazzoni, G.; Farinetti, Alberto; Sabatini, A. M.; Nanni, N.; Grottola, Antonella; Sgambato, A.
abstract

...

2011 - Lymph node micrometastasis and survival of patients with Stage I (Dukes' A) colorectal carcinoma. [Articolo su rivista]
REGGIANI BONETTI, Luca; Di Gregorio, C; DE GAETANI, Carmela; Pezzi, A; Barresi, G; Barresi, V; Roncucci, Luca; PONZ DE LEON, Maurizio
abstract

Objective. Although patients with Stage I colorectal cancer show an excellent prognosis, a few of them die of metastatic disease. In this subgroup of individuals, the search of occult metastasis might reveal that early dissemination of tumor cells could be the cause of cancer progression. Material and methods. Through a Cancer Registry, we selected all patients with Stage I disease who died of metastatic tumor; a total of 32 patients were identified and in 25 of them paraffin-embedded material was available. The group was matched to 70 Stage I patients with favorable prognosis (controls). In cases and controls resected lymph nodes were cut, and micrometastases were searched using pan-cytokeratin antibodies. Results. Micrometastases were detected in 18 of 25 (72%) Stage I patients who died of the disease, while they were almost absent among controls (1 of 70, p < 0.001 by χ2 test). Vascular invasion and tumor budding were more frequent among Stage I patients with an unfavorable prognosis than in controls. By regression analyses, micrometastases (HR 12.3, CI 4.8–32) and vascular invasion (HR 3.5, CI 1.4–8.5) maintained an independent association with prognosis (cancer-specific survival). Conclusion. Micrometastasis in the lymph nodes can be revealed in the majority of patients with early colorectal cancer who die of tumor progression, while they appear extremely rare in Stage I individuals with good prognosis. The selection of patients through histology (vascular invasion) and search of occult metastatic cells might represent a way to identify individuals who might benefit from adjuvant chemotherapy.

2011 - Lymphatic vessel density and its prognostic value in stage I colorectal carcinoma. [Articolo su rivista]
Barresi, V; Reggiani Bonetti, L; Di Gregorio, C; PONZ DE LEON, Maurizio; Barresi, G.
abstract

AIMS:The assessment of lymphatic vessel density (LVD) has been suggested as a tool to determine the metastatic risk of neoplasias. On this premise, the authors aimed to verify whether progression risk of stage I colorectal cancer may be related to LVD. The authors also evaluated and correlated vascular endothelial growth factor (VEGF)-A expression with LVD revealed in the same cases in order to investigate its potential lymphangiogenic role in the early stage colorectal cancer.METHODS:LVD and VEGF immunoexpression were analysed and compared in series of 29 stage I surgically resected colorectal carcinomas obtained from patients showing disease progression and in a cohort of 23 stage I colorectal cancers from patients with no evidence of disease progression. The prognostic value of LVD and of VEGF expression on the progression-free survival to colorectal cancer was investigated.RESULTS:A high density of peritumoural lymphatics (P-LVD) was significantly associated with high VEGF expression and disease progression. Moreover, high P-LVD and high VEGF expression were significant negative prognostic parameters associated with a shorter disease-free interval in stage I colorectal cancer.CONCLUSIONS:If our findings are further confirmed in other studies, the assessment of P-LVD on surgical specimens might be used as a tool to identify patients with stage I colorectal cancer at higher risk of progression in order to submit them to adjuvant therapies. Since P-LVD seems to show a VEGF-A mediated regulation in stage I colorectal cancer, therapies targeting this factor might be exploited to reduce lymphangiogenesis and the progression risk of this neoplasia.

2011 - Morphological analysis of three extrathoracic bronchogenic cysts simulating neoplasms [Articolo su rivista]
Lupi, M.; Reggiani Bonetti, L.; Stauder, E.; Bettelli, S.; Maiorana, A.
abstract

Bronchogenic cyst is a congenital anomaly of the primitive foregut. Unusual occurrences in extrathoracic sites have been described in the literature, some of which may clinically simulate neoplasms. We report three additional cases arising near the left adrenal gland, left ovary and gastric wall. Pathological findings are discussed, together with a review of the literature.

2011 - Neutrophil gelatinase-associated lipocalin (NGAL) and matrix metalloproteinase-9 (MMP-9) prognostic value in stage I colorectal carcinoma. [Articolo su rivista]
Barresi, V; Reggiani Bonetti, L; Di Gregorio, C; Vitarelli, E; PONZ DE LEON, Maurizio; Barresi, G.
abstract

The expression of neutrophil gelatinase-associated lipocalin (NGAL) has been suggested to behave likea negative prognostic marker in stage I colorectal carcinoma. In the aim of clarifying whether its associationwith adverse outcome may descend from NGAL’s ability to regulate matrix metallo-proteinase-9(MMP-9), we analyzed the correlation, prognostic value, and association with neo-angiogenesis of NGALand MMP-9 immunohistochemical expression in a series of stage I colorectal carcinomas. A variableNGAL immunoexpression was demonstrated in 17 of the 48 analyzed cases with a significantly higherfrequency of positive cases among patients showing disease progression. NGAL expression was also positivelycorrelated with VEGF expression detected in the same cases. MMP-9 immunostaining was presentin the cytoplasm of the neoplastic cells in 30 cases; no significant correlations were evidenced with NGALexpression, as well as with the various clinico-pathological parameters or with progression of the colorectalcarcinomas. By contrast, NGAL expression was confirmed as a significant independent negativeprognostic marker related to a shorter disease-free survival in stage I colorectal carcinoma. Our preliminaryresults suggest that the association of NGAL with poor outcome might be independent from MMP-9regulation, thus highlighting its prognostic value in this neoplasia. If our findings are confirmed in furtheranalyses, NGAL assessment might be used in order to select those patients with a higher progression riskand to submit them to adjuvant therapies useful to prevent adverse outcome.

2011 - Neutrophil gelatinase-associated lipocalin: A new prognostic marker in stage i colorectal carcinoma? [Articolo su rivista]
Barresi, Valeria; Di Gregorio, Carmela; Reggiani-Bonetti, Luca; Ieni, Antonio; Ponz-De Leon, Maurizio; Barresi, Gaetano
abstract

TNM stage I colorectal cancer is commonly characterized by a good prognosis, with 5-year survival of around 80% to 90%. Nonetheless, disease progression occurs in a percentage of cases, although the causes of an adverse clinical course still remain to be clarified. In the present study, we analyzed and compared the immunohistochemical expression of neutrophil gelatinase-associated lipocalin, an iron-binding protein, which is involved in colorectal cancer progression, in series a of 29 surgically resected colorectal carcinomas obtained from patients who died of the disease and in a cohort of 22 colorectal cancers from patients alive 5 years after the initial diagnosis. The prognostic value of neutrophil gelatinase-associated lipocalin expression on the overall survival to colorectal cancer was investigated. Variable neutrophil gelatinase-associated lipocalin immunoexpression was demonstrated in 23 of the 51 analyzed cases, with a significantly higher frequency of positive cases among patients who died of the disease. Moreover, neutrophil gelatinase-associated lipocalin expression appeared to be a significant independent negative prognostic marker related to shorter overall survival in stage I colorectal carcinoma. If our findings are confirmed in further analyses, neutrophil gelatinase-associated lipocalin assessment might be used to select patients with a higher risk of progression and to find adjuvant therapies for the prevention of adverse outcomes. Â© 2011 Elsevier Inc.

2010 - Atypical lymphoid proliferation in capillary hemangioma: A finding related to bacterial infection? [Articolo su rivista]
Cesinaro, Anna M.; Reggiani Bonetti, L
abstract

not available

2010 - Cerebrospinal fluid cytology in a case of primary diffuse leptomeningeal and pineal melanocytic lesion, with histological confirmation [Articolo su rivista]
Zunarelli, Elena; Bettelli, Stefania; Reggiani-Bonetti, Luca; Bertolini, Federica; Falasca, Angelo
abstract

Non available

2010 - EGFR polysomy in squamous cell carcinoma of the thyroid. Report of two cases and review of the literature [Articolo su rivista]
Bonetti, L. R.; M., Lupi; M., Trani; N., Trani; Sartori, Giuliana; Schirosi, Laura; Bettelli, Stefania Raffaella; G., Zanelli; L., Maccio; Maiorana, Antonino
abstract

AIMS AND BACKGROUND:Primary squamous cell carcinoma of the thyroid gland (PSCCT) is an uncommon malignancy characterized by a poor prognosis. A radical surgical approach combined with radiotherapy or chemotherapy is the generally accepted treatment for this tumor. The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor modulating the cell proliferation and biological progression of many human epithelial tumors. The EGFR overexpression in PSCCT suggests an additional therapeutic option for the treatment of this tumor.METHODS AND STUDY DESIGN:The clinicopathological features and immunohistochemical profiles of two cases of primary squamous cell carcinoma of the thyroid in a 66-year-old and an 83-year-old woman are presented. EGFR status was valued in both cases.RESULTS:Overexpression of EGFR protein was detected in 50% and 75% of the tumor cell membranes. EGRF gene polysomy was detected in both tumors.CONCLUSIONS:Pharmaceuticals targeting EGFR may help to provide the rationale for an additional, novel therapeutic option for this rare tumor, especially when other therapeutic options have been exhausted.

2010 - Liver injury by experimental portal bacteremia: histogenetic recovery study in the rat [Working paper]
Manenti, A.; Trenti, L.; Reggiani Bonetti, L.; Zaffe, Davide; Botticelli, A.
abstract

Abstract - To study the histogenetic recovery of hepatic lesions due to portal bacteremia, a complication of some clinical conditions, an experimental animal model had developed. Portal bacteremia was performed in 8-week rats and the morphological recovery of liver was histologically checked 1 to 6 days after bacteria inoculation. The major injuries, such as acute inflammatory exudate of the portobiliary spaces, piecemeal necrosis of muralium, micro-abscesses and areas of hepatocyte necrosis of the liver parenchyma, and thrombosis in the centrolobular vein were recorded 1 day after inoculation. Minimal signs of vacuolar degeneration, steatosis, necrosis areas, vessel congestion and focal hemosiderosis together with a small hepatocyte proliferative activity was instead appreciable with longer time. The results seem to suggest a role of vascular structures and Kupffer cells in the morphological repair. This experimental model could serve to understand better similar clinical hepatology conditions, such as portal bacteremia.Informazioni utili

2010 - Stage I colorectal carcinoma: VEGF immunohistochemical expression, microvessel density, and their correlation with clinical outcome. [Articolo su rivista]
Barresi, V; Di Gregorio, C; Reggiani Bonetti, L; PONZ DE LEON, Maurizio; Barresi, G; Vitarelli, E.
abstract

Tumor-node-metastasis (TNM) stage I colorectal cancer is commonly characterized by a good prognosis, with 5-year survival around 80-90%; nonetheless, it undergoes disease progression in a percentage of cases, although the causes of adverse clinical course still remain to be elucidated. In the present study, we analyzed and compared the immunohistochemical expression of the pro-angiogenic vascular endothelial growth factor (VEGF) as well as the microvessel density (MVD) in a series of 27 surgically resected colorectal carcinomas obtained from patients deceased because of disease progression and in a cohort of 25 colorectal cancers from patients still alive with no evidence of disease progression 5 years after the initial diagnosis. The prognostic value of VEGF expression and of MVD on the overall survival to colorectal cancer was investigated. A variable VEGF immunoexpression was demonstrated in all the analyzed cases. High VEGF expression was significantly more frequent among patients deceased of the disease. These patients also displayed significantly higher MVD counts in their cancer in comparison to the patients alive after 5 years from surgery. Moreover, both high VEGF expression and MVD appeared as significant negative prognostic markers related to a shorter overall survival to stage I colorectal carcinoma, with VEGF representing an independent variable at multivariate analysis. VEGF assessment might be used in order to select those patients with a higher progression risk and to submit them to adjuvant therapies useful to prevent adverse outcome.

2010 - The role of fetal autopsy and placental examination in the causes of fetal death: a retrospective study of 132 cases of stillbirths. [Articolo su rivista]
Bonetti, L. R.; P., Ferrari; N., Trani; L., Maccio; S., Laura; S., Giuliana; Facchinetti, Fabio; Rivasi, Francesco
abstract

PURPOSE: To investigate the most plausible cause of stillbirth by evaluating clinical records and postmortem examination findings including placental analysis. METHODS: A retrospective cohort study concerning 132 stillbirths from 124 pregnancies occurred in the Mother-Infant Department of the University Hospital of Modena, Italy, from January 2000 to December 2004. Collected data were reviewed and classified according to the Gardosi ReCoDe system. RESULTS: A reasonable cause of fetal death was identified in 99/124 pregnancies (79.84\%). No associated relevant factors were disclosed in 25 fetuses (20.16\%) classified as unexplained stillbirths. A succeeding scrupulous analysis of the placenta and an accurate clinical record review were useful to detect other conditions in 82 cases, including 5 cases of unexplained stillbirth. The major relevant conditions associated to stillbirths were feto-placental infection especially in the early fetal gestation age, under the 24th week of gestation, and placental insufficiency occurred both in early and late gestation age fetuses and mainly associated with a IUGR (<10th customized percentile). The main frequent secondary conditions were represented by placental anomalies including cluster of avascular villi with stromal fibrosis associated to thrombosis in minor and/or major vessel(s). Through the further analysis of the placenta, we were able to reduce the unexplained stillbirth rate from 20.16 to 15\%. CONCLUSION: Accurate fetal autopsy and placental examination related to meticulous clinical collecting data are requisites in the valuation of stillbirth and could play an important role in reduction of unexplained stillbirth rate.

2009 - Congenital tracheal atresia in newborn: case report and review of the literature [Articolo su rivista]
Lupi, M.; L. R., Bonetti; N., Trani; Maccio, Livia; Maiorana, Antonino
abstract

Tracheal atresia is an uncommon congenital malformation with a high mortality rate. Clinical symptoms occur suddenly after birth. The diagnosis is suspected in any infant in whom improved ventilation is obtained despite aggressive attempts at resuscitation. We describe a small gestational week 34 male newborn affected by tracheal atresia without esophageal fistula with associated fetal growth restriction, ascites and polyhydramnios. Post mortem examination revealed a diffuse cyanotic status, abdominal ascites and a low birth weight. A 3 cm tract of trachea was documented that distantly ended in a blind pouch and without tracheoesophageal fistulae and enlarged bulky lungs connected to each other by a common thin-walled bronchus. Histological examination showed a normal conformed larynx and scratchily cartilaginous disks in the proximal tract of the short trachea. Vascular space referred to small arteries and veins, thin bands of fibrous tissue and adipose tissue were detected under the blind pouch. Lung distal airspaces were lined by premature cubic epithelium separated by a broad poorly vascularized interstitium. A striking interstitial and alveolar edema was remarkable.

2009 - Expression of estrogen receptor in hemangioma of the uterine cervix: Reports of three cases and review of the literature [Articolo su rivista]
Reggiani Bonetti, Luca; Boselli, F.; Lupi, M.; Bettelli, S.; Schirosi, L.; Bigiani, N.; Sartori, G.; Rivasi, Francesco
abstract

The occurrence of hemangioma in the female genital tract, particularly in uterine cervix, is rare. The majority of them show asymptomatic behavior. Surgical excision remains curative in most of the cases. Conservative therapies such as sclerosing agents, cryotherapy, and CO(2) laser excision may be alternatively applied. We present three cases of hemangiomas of the cervix in asymptomatic women, diagnosed as cavernous hemangioma in two cases and capillary hemangioma in one. All tumors were immunoreactive for CD31, CD34, factor-VIII-related antigen. Focal expression of estrogen receptors was detected. No positivity was obtained with progesterone receptor antibodies. The presence of estrogen receptor in the endothelial cells of the hemangioma of the cervix suggests a direct role of this hormone in the hemangioma development. A possible target therapy is discussed.

2009 - Giant genital cavernous haemangioma: case description and surgical management. [Articolo su rivista]
Sighinolfi, Maria Chiara; Bonetti, L. R.; DE STEFANI, Stefano; M., Pinelli; Micali, Salvatore; Maiorana, Antonino; L., Schirosi; G., Sartori; DE GAETANI, Carmela; Bianchi, Giampaolo
abstract

Giant genital haemangiomas are rare occurrences. Once properly diagnosed, they should be managed by surgery with wide and deep margins. We present a clinical case and provide suggestions for diagnosis and treatment of this unusual pathology.

2009 - Ovarian teratoma predominantly consisting of thymic tissue: Case report and review of the literature [Articolo su rivista]
Rossi, Giulio; Reggiani-Bonetti, Luca; Ventura, Vito; De Martis, Stefano; Catani, Augusto
abstract

Non available

2009 - Rapidly-growing hemangioma of the testicle clinically simulating an aggressive neoplasm. A case report [Articolo su rivista]
Bonetti, L. R.; Schirosi, Laura; Sartori, Giuliana; Lupi, Massimo; DE GAETANI, Carmela; N., Trani; Maiorana, Antonino
abstract

We present a case of intraparenchymatous capillary-type hemangioma of the testicle in an adult. The patient was a 37-year-old man who showed a rapidly enlarging and palpable mass in left testicle. Radical orchiectomy was performed, and histological examination revealed an unencapsulated lobulated tumour with wide hemorrhagic portions. To our knowledge, the occurrence of rapid enlargement in a testicular hemangioma has not been previously reported, which might be explained by the development of intra-tumoural haemorrhage.

2009 - Relative role of APC and MUTYH mutations in the pathogenesis of familial adenomatous polyposis. [Articolo su rivista]
Pezzi, A; Roncucci, Luca; Benatti, Piero; Sassatelli, R; Varesco, L; Di Gregorio, C; Venesio, T; Pedroni, Monica; Maffei, S; Reggiani Bonetti, L; Borsi, E; Ferrari, M; Martella, P; Rossi, G; PONZ DE LEON, Maurizio
abstract

OBJECTIVE: Familial adenomatous polyposis (FAP) is an interesting model for the study of colorectal tumour. Two genes contribute to the FAP phenotype - APC and MUTYH - but their relative role is still undefined. The objective of this study was to evaluate the contribution of the two genes to the pathogenesis of FAP by means of a series of FAP families. MATERIAL AND METHODS: Sixty-one unrelated families with a diagnosis of FAP and a total of 187 affected individuals were evaluated. After extracting DNA, APC and MUTYH genes were sequenced. RESULTS: In the whole series of patients, colectomy with ileorectal anastomosis was the most frequent surgery, although the number of patients treated by total proctocolectomy and ileoanal anastomosis was increasing. Duodenal and jejunal-ileal adenomas were present in more than half of the patients. Constitutional mutations were detected in 37 of the 45 families (82.2%); there were 33 families with APC and 4 with MUTYH alterations. Age at onset of polyposis and age at surgery were 10-15 years delayed for carriers of MUTYH mutations; cancer at diagnosis was frequent, and extracolonic manifestations were diagnosed in the majority of MUTYH-positive families. MUTYH-associated polyposis showed the horizontal transmission expected for recessive inheritance (at variance with the dominant pattern seen with APC mutations). CONCLUSIONS: At least two genes are associated with the FAP phenotype. APC mutations account for the majority of cases, while MUTYH mutations can be observed in 10% of patients. There are few but definite differences between APC- and MUTYH-associated FAP, such as age at diagnosis and pattern of transmission.

2008 - Hyperplastic and inflammatory pathology of the thymus [Capitolo/Saggio]
Maiorana, Antonino; REGGIANI BONETTI, L.
abstract

n/a

2008 - Immediate anaphylactic death following antibiotics injection: Splenic eosinophilia easily revealed by pagoda red stain [Articolo su rivista]
Trani, Nicoletta; Bonetti, Luca Reggiani; Gualandri, Giorgio; Barbolini, Giuseppe
abstract

The comparison of 4 cases of immediate anaphylactic death following the intramuscular injection of antibiotics (different types of penicillin or cephalosporins) with 4 cases of immediate non-anaphylactic death (induced by different causes) recognized splenic eosinophilia, as the main feature for the differential diagnosis, in agreement with isolated previous studies. The use of a stain (pagoda red) little known and seldom employed in Pathology and in Forensic Medicine, showed the concomitant massive presence (immunohistochemically confirmed) of mast cells and degranulated mast cells, the latter mainly located in splenic sinuses. The whole of our findings led us to consider the spleen as the possible shock organ in man. Waiting for further judgment on our hypothesis, we seized the opportunity to remark the employment of the pagoda red stain when the contemporary demonstration of eosinophils, mast cells and degranulated mast cells is required. Â© 2008 Elsevier Ireland Ltd. All rights reserved.

2008 - Well-differentiated giant scrotal liposarcoma: case presentation and management [Articolo su rivista]
Sighinolfi, M. C.; DE STEFANI, Stefano; Micali, Salvatore; G., Saredi; F., Fidanza; A., Dotti; L., Reggiani Bonetti; C., De Gaetani; Bianchi, Giampaolo
abstract

Scrotal liposarcoma is an uncommon disease, usually found after the fifth decade. We describe the case of a well-differentiated scrotal liposarcoma associated with a considerable inflammatory reaction, treated with surgical ablation.

2006 - Perinatal aspergillosis: a case with fatal outcome. [Articolo su rivista]
Rivasi, Francesco; Cesinaro, Am; Curatola, C; REGGIANI BONETTI, Luca
abstract

Aspergillosis is an uncommon perinatal infection diagnosed with increasing frequency in recent years. We report a premature infant who required both nutrition and ventilation artificially assisted and developed a disseminated invasive nosocomial infection from Aspergillus flavus. Autopsy revealed marked hypotrophy of the thymus and multisystem invasive aspergillosis chiefly involving the vascular and alimentary systems and also the respiratory tract, the central nervous system, and the skin. From what we know, this is the first case of the literature with a misleading initial clinical presentation involving the alimentary tract (hepatomegaly, ingravescent cholestatic icterus) and evolving in intestinal occlusion.

2003 - Regression of hypertrophy influences ventricular performance in patients treated with telmisartan [Abstract in Atti di Convegno]
Mattioli, Av; Bonatti, S; Bonetti, L; Zennaro, M; Monopoli, D; Mattioli, G
abstract

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Università degli studi di Modena e Reggio Emilia

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