Nuova ricerca

Mario MIGALDI

Ricercatore Universitario
Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con interesse Trapiantologico, Oncologico e di Medicina Rigenerativa

Home | Curriculum(pdf) | Didattica |


Pubblicazioni

2023 - Changes in coffee and caffeine intake during the pandemic in women smokers and non-smokers: a future challenge for cardiovascular prevention [Articolo su rivista]
Coppi, Francesca; Migaldi, Mario; Stefanelli, Claudio; Farinetti, Alberto; Mattioli, Anna Vittoria
abstract

Background: The recent pandemic has led to major lifestyle changes, especially in women, changes that will impact cardiovascular risk. The aim of the present observational study was to evaluate changes occurred during pandemic in coffee and caffeine intake in a group of adult women and compare changes in smoking versus non-smoking women. Methods: A web questionnaire was sent through a online survey platform to a group of unselected adult women. The consumption of coffee and caffeine were investigated in 2 groups of women by comparing smokers and non-smokers. Results: A total of 435 adult women (256 non-smokers and 179 smokers) answer to all questions. Smokers increase the number of cigarette/days (mean + 3.4 cig/day). Coffee intake was significantly increase in smokers compared to non-smokers (3.1+1.0 versus 1.5+0.6 cups/day p<0.01). In smokers, self-perception of increase stress was related to increased coffee intake (r = 0.84; p <0.001), increased sugar- rich foods (r=0.81; p<0.001), increased chocolate rich snacks (r=0.72; p<0.01), increased sitting time (r=0.79; p<0.01). Conclusions: These preliminary data must suggest to undertake social campaigns aimed at encouraging a return to a healthy lifestyle that certainly includes a healthy diet but also the suspension of smoking. These observational results need further evaluation with prospective studies in order to quantify the effects of pandemic-induced changes in lifestyle on cardiovascular risk in women.

2022 - Cutaneous Melanoma Systematic Diagnostic Workflows and Integrated Reflectance Confocal Microscopy Assessed with a Retrospective, Comparative Longitudinal (2009–2018) Study [Articolo su rivista]
Pellacani, G.; Farnetani, F.; Chester, J.; Kaleci, S.; Ciardo, S.; Bassoli, S.; Casari, A.; Longo, C.; Manfredini, M.; Cesinaro, A. M.; Giusti, F.; Iacuzio, A.; Migaldi, M.
abstract

Background: The increasing global burden of melanoma demands efficient health services. Accurate early melanoma diagnosis improves prognosis. Objectives: To assess melanoma prevention strategies and a systematic diagnostic-therapeutical workflow (improved patient access and high-performance technology integration) and estimate cost savings. Methods: Retrospective analysis of epidemiological data of an entire province over a 10-year period of all excised lesions suspicious for melanoma (melanoma or benign), registered according to excision location: reference hospital (DP) or other (NDP). A systematic diagnostic-therapeutical workflow, including direct patient access, primary care physician education and high-performance technology (reflectance confocal microscopy (RCM)) integration, was implemented. Impact was assessed with the number of lesions needed to excise (NNE). Results: From 40,832 suspicious lesions excised, 7.5% (n = 3054) were melanoma. There was a 279% increase in the number of melanomas excised (n = 203 (2009) to n = 567 (2018)). Identification precision improved more than 100% (5.1% in 2009 to 12.0% in 2018). After RCM implementation, NNE decreased almost 3-fold at DP and by half at NDP. Overall NNE for DP was significantly lower (NNE = 8) than for NDP (NNE = 20), p < 0.001. Cost savings amounted to EUR 1,476,392.00. Conclusions: Melanoma prevention strategies combined with systematic.

2021 - Subclinical Atherosclerosis at Peripheral Arteries in Obese Individuals. [Articolo su rivista]
Farinetti, A; Castaniere, I; Clini, E; Migaldi, M; Gelmini, R; Scaringi Raspagliesi, F; Ara, N; Serra, F; Spatafora, F; Genazzani, A; Mattioli, Av.
abstract

Evidence on relationship between obesity and peripheral arterial disease (PAD) are controversial. The aim of the present study is to evaluate the presence of subclinical atherosclerosis at all level of the explorable vascular segments with a systematic method in a selected population of young obese submitted to a comprehensive rehabilitation course. A group of 50 consecutive morbidly obese (BMI>30) was included. All patients underwent Doppler evaluation including intima media thickness (IMT) and presence/absence of plaques. We found that vessels in the upper segment of the body demonstrate the presence of thickening and/or plaques at the level of the carotid segments but not of the subclavian arteries. The IMT of the right Common Carotid Artery (CCA) (1.49 + 1.38 versus 0.62 + 0.23; p=0.037) and of the left CCA (1.66 + 1.89 versus 0.45 + 0.26; p=0.034) was greater in patients. Vessels of the lower segment demonstrate the presence of thickening and/or plaques at the iliac but not at femoral level. The control group did not present vessel thickening at any level. In conclusions asymptomatic vascular damage may be present in different segment of peripheral vessels, thus suggesting an early risk for developing an overt vascular disease over time in obese.

2020 - Atypical fibroxanthoma and pleomorphic dermal sarcoma: A reappraisal [Articolo su rivista]
Cesinaro, A M; Gallo, G; Tramontozzi, S; Migaldi, M
abstract

Atypical fibroxanthoma (AFX) and pleomorphic dermal sarcoma (PDS) share clinical, pathological, immunohistochemical and molecular features, though PDS is associated with a more aggressive behavior.

2020 - Expression of calretinin in odontogenic keratocysts and basal cell carcinomas: A study of sporadic and Gorlin-Goltz syndrome-related cases. [Articolo su rivista]
Cesinaro, Am; Burtini, G; Maiorana, A; Rossi, G; Migaldi, M.
abstract

Gorlin-Goltz syndrome (GGS), is an autosomal dominant inherited disorder related to germline mutation of PTCH1 gene, characterised by the presence of multiple developmental anomalies and tumours, mainly basal cell carcinomas (BCC) and odontogenic keratocysts (OKC). We analysed and compared the expression of calretinin in 16 sporadic OKCs, from 15 patients, and 12 syndromic OKCs from 11 patients; in 19 BCC's and 2 cutaneous keratocysts (CKC) belonging to 4 GGS patients, 15 sporadic BCCs and 3 steatocystomas (SC). Calretinin was negative in 10 of 12 syndromic OKCs, focally positive (<5% of cells) in 2; six sporadic OKCs were negative, 6 focally and 4 diffusely positive (p = .02, cases focally and diffusely positive vs. cases negative). All BCCs of 3 GGS patients were negative, the fourth patient presented two BCCs negative and 5 focally or diffusely positive; 7 sporadic BCCs were negative and 8 focally positive (p = NS). Two CKCs resulted negative in one GGS patient; 2 sporadic SCs were positive, and a third was negative. PTCH1 mutations produce an altered PTCH protein and an aberrant activation of Sonic hedgehog (SHH) pathway, leading to tumoral proliferation. It has been demonstrated that treatment of human foetal radial glia cells with SHH reduces, whereas the blockage of SHH increases calretinin expression. We found a lower expression of calretinin in syndromic OKCs compared to sporadic cases. Although calretinin's value in differential diagnosis between sporadic and syndromic tumours appears not crucial, our results shed light on the possible link between SHH dysfunction and calretinin expression in GGS-related tumours.

2019 - Subclinical atherosclerosis at peripheral arteries in obese subjects [Abstract in Rivista]
Farinetti, Alberto; Clini, Enrico; Migaldi, Mario; Gelmini, Roberta; Serra, Francesco; Spatafora, Francesco; Scaringi Raspagliesi, Flavia; Ara, Nicoletta
abstract

2019 - THE CLINICAL AND FORENSIC ROLE OF CITOLOGY IN PTA AND PT1 BLADDER CANCER MONITORING. CASE STUDY REVISION FOR THE PERIOD 2008 – 2017. [Articolo su rivista]
9., Francesco Raschellà; Marella Gian, Luca; Potenza, Saverio; Migaldi, Mario; Caggiano, Bartolo; Marsella, Luigi; Tavone, Alessandro; Iacuzio, Antonio
abstract

Introduction: The Authors describe the results of a retrospective study that analyzes importance of a proper bladder cancer monitoring, comparing the use of the different methods available, both in terms of diagnostic delay and in terms of legal medical repercussions. Materials and methods: Using the database of the Pathological Anatomy Department of the Modena Polyclinic, we have isolated a series of 238 patients with histological diagnosis of bladder urothelial carcinoma in pTa and pT1 stages with an observational minimum time interval after first diagnosis of at least 5 years. The observational statistical analysis of the data stored was made through a statistical software (SPSS report 11.00 USA). Results: The results of the present study show how cytological screening, performed constantly with urine tests during early-stage monitoring of bladder tumors, can be a valid tool for the timely diagnosis of tumor stage evolution. Indeed positivity of the cytological examination can direct to a rapid diagnostic and therapeutic re-planning. Conclusion: It would be desirable to standardize the best screening strategies about bladder cancer. With a correct standardization, a valid reference could be obtained both from a clinical point of view, and for a correct legal medical evaluation in term of diagnostic delay and, consequently, reduction in the chance of survival.

2018 - Cardiac involvement in systemic sclerosis: identification of high-risk patient profiles in different patterns of clinical presentation [Articolo su rivista]
Coppi, Francesca; Giuggioli, Dilia; Spinella, Amelia; Colaci, Michele; Lumetti, Federica; Farinetti, Alberto; Migaldi, Mario; Rossi, Rosario; Ferri, Clodoveo; Boriani, Giuseppe; Mattioli, Anna Vittoria
abstract

Systemic sclerosis (SSc) is a chronic connective tissue disease characterized by widespread microvascular damage, dysregulation of fibroblasts with collagen overproduction and excessive fibrosis of the skin and internal organs, as well as complex immune system abnormalitie….

2018 - Coffee in hypertensive women with asymptomatic peripheral arterial disease: a potential nutraceutical effect [Articolo su rivista]
Mattioli, Anna V; Migaldi, Mario; Farinetti, Alberto
abstract

Several studies suggest that coffee is associated with a lower risk of atherosclerosis.1,2 Coffee contains polyphenol antioxidants, which have been hypothesized to act against free oxygen radicals and lipid peroxidation and to improve endothelial function.1–3 Coffee is part of the Mediterranean diet, one of the healthiest diets. The characteristics.....

2018 - Diffuse Idiopathic Pulmonary Neuroendocrine Cell Hyperplasia (DIPNECH) Syndrome and Carcinoid Tumors With/Without NECH: A Clinicopathologic, Radiologic, and Immunomolecular Comparison Study. [Articolo su rivista]
Mengoli, Mc; Rossi, G; Cavazza, A; Franco, R; Marino, Fz; Migaldi, M; Gnetti, L; Silini, Em; Ampollini, L; Tiseo, M; Lococo, F; Fournel, L; Spagnolo, P; Cottin, V; Colby, Tv.
abstract

The diagnostic criteria of diffuse idiopathic pulmonary neuroendocrine cell hyperplasia (DIPNECH) are not well defined, and DIPNECH can be mistaken for carcinoids associated with neuroendocrine cell hyperplasia (NECH). In this study, we compared clinical, radiologic, histologic, immunohistochemical, and molecular features of DIPNECH and isolated carcinoids with/without NECH. The study population included 151 cases (77 female patients and 74 male patients), 19 with DIPNECH and 132 with carcinoids with/without NECH. None of the cases displayed molecular alterations or anaplastic lymphoma kinase expression. Compared with individuals with carcinoids with/without NECH, patients with DIPNECH were more likely to be female individuals (P<0.0001), nonsmokers (P=0.021), and symptomatic, and to have an obstructive/mixed respiratory defect, peripheral location of the lesions, and air trapping (P<0.0001) on chest computed tomography, and constrictive bronchiolitis on histology (P<0.0001). Among immunohistochemical markers, DIPNECH was associated with higher expression of thyroid transcription factor-1, CD10, and gastrin-releasing peptide/bombesin-like peptide (P<0.0001). Yet, when a purely histopathologic definition of DIPNECH was applied, 40% of isolated carcinoids also met the diagnostic criteria for DIPNECH, even in the absence of symptoms and/or radiologic abnormalities. Therefore, as DIPNECH represents a distinct clinical syndrome, we suggest the term DIPNECH be limited to cases presenting with respiratory symptoms, functional and/or radiologic abnormalities, and constrictive bronchiolitis on histology.

2018 - Fruit and vegetables in hypertensive women with asymptomatic peripheral arterial disease [Articolo su rivista]
Mattioli, Av; Coppi, F; Migaldi, M; Farinetti, A.
abstract

Background and aims: Fruit and vegetables are considered a very healthy diet useful in the prevention of cardiovascular disease. The present study aims to evaluate intake of fruit and vegetables in hypertensive women and its correlation with asymptomatic atherosclerosis. Methods and results: A group of 237 women with hypertension was evaluated. Fruit and vegetables consumption were assessed by a self-administered food frequency validated questionnaire completed by an interviewer administered 24 h diet recall. They all underwent ABI. ABI measurement observed that fruit consumption was inversely associated with pre-clinical atherosclerosis suggesting a protective effect, moreover this association was stronger for vegetables. Increasing intake of vegetables was associated with a lower risk of asymptomatic PAD. Conclusions: Women with a high intake of fruit and vegetables showed less instrumental sign of preclinical peripheral atherosclerosis. Can be suggests that fruit and vegetables play an important role in prevention of atherosclerosis in pre-menopausal women.

2018 - Impact of Dermoscopy and Reflectance Confocal Microscopy on the Histopathologic Diagnosis of Lentigo Maligna/Lentigo Maligna Melanoma [Articolo su rivista]
Mataca, E; Migaldi, M; Cesinaro, Am.
abstract

BACKGROUND: Equivocal pigmented lesions of the head are usually biopsied to avoid inappropriate treatment. Clinical approach has evolved from simple visual examination to sophisticated techniques for selecting the biopsy sites. OBJECTIVE: This study aimed to retrospectively evaluate the efficiency of dermoscopy (DE) and reflectance confocal microscopy (RCM) in sampling a histopathologically representative focus of lentigo maligna/lentigo maligna melanoma. METHODS: Punch biopsies and surgical excisions of 72 patients, 37 men and 35 women (median age 70.6 years, range 39-90 years), affected by lentigo maligna/lentigo maligna melanoma of the head, sent from a single dermatology clinic, were reviewed for the presence of 5 histopathologic criteria: atypical junctional melanocytes, increased junctional melanocytes, follicular colonization, pagetoid spread and melanocytic junctional nests, plus other minor features. Forty-two patients were biopsied under DE and 30 under RCM guidance. RESULTS: Accuracy of the 2 techniques in sampling a representative tissue overlapped in most cases, although RCM selected sites to biopsy with more histopathologic criteria, in particular pagetoid spread and melanocytic nests. Interestingly, with RCM, inflammation and melanophages were observed more in biopsy than in excision. False positive cases were not registered. CONCLUSION: Compared with the sampling at naked eye, our results show that DE and RCM help selecting the most appropriate areas for biopsies, thus allowing not only more robust histopathologic diagnoses, but also a more accurate microstaging of tumor.

2018 - Physical activity in premenopausal women with asymptomatic peripheral arterial disease [Articolo su rivista]
Mattioli, Av; Coppi, F; Migaldi, M; Farinetti, A
abstract

...

2018 - The diagnostic delay of oral carcinoma [Articolo su rivista]
Marella Gian, Luca; Raschellà, Francesco; Solinas, Matteo; Mutolo, Pietro; Potenza, Saverio; Milano, Filippo; Mauriello, Silvestro; Caggiano, Bartolo; Rondinelli, Pierfrancesco°; Anesi, Alexandre°°; Migaldi, Mario°
abstract

This paper describes the results of a retrospective study that analyzed the extent and role of diagnostic delays on the development and prognosis of oral cancer. We consulted the digital archives of the Anatomy and Pathology Department of the University Hospital of Modena and Reggio Emilia for the period from 2000 to 2016, to identify all patients with oral cavity lesions according to the SNOMED coding system. In total, 645 reports of squamous cell carcinoma of the oral cavity were retrieved. Data collected from the reports was supplemented with clinical information, with particular reference to the time of onset of the first signs and/or symptoms and the time elapsed between biopsy and definitive histological diagnosis following surgery. The average delay of patients from onset of signs and/or symptoms and seeking medical care was 112 days, or about 4 months. A longer delay was found for male with respect to female patients (151 days versus 82 days respectively; p < 0.015). An average delay of 40 days was observed between the first biopsy and the postoperative histological diagnosis. Results indicate that diagnostic delays occur frequently in oral tumours and can due to both the patient's wait-and-see conduct upon appearance of the first signs and/or symptoms, and to the organizational and communicative deficiencies among the different medical specialties. In light of these results, we make the recommendation to organize information campaigns through the Public Health Departments and specific screening programs, and to introduce an operational protocol for the prevention and early diagnosis of oral cancer, involving general medicine practitioners and dentists as the main promoters.

2017 - Analysis of a panel of druggable gene mutations and of ALK and PD-L1 expression in a series of thymic epithelial tumors (TETs). [Articolo su rivista]
Tiseo, M; Damato, Angela; Longo, Luigia; Barbieri, F; Bertolini, F; Stefani, Alessandro; Migaldi, M; Gnetti, L; Camisa, R; Bordi, P; Buti, S; Rossi, G
abstract

Introduction: Thymic epithelial tumors (TETs) are rare neoplasms with different prognosis lacking consis-tent molecular alterations possibly leading to targeted therapy. We collected a consecutive series of TETsaimed at investigating the mutational status of druggable genes (EGFR, c-KIT, KRAS, BRAF, PDGFR-alphaand −beta, HER2 and c-MET) and the expression of ALK and PD-L1.Patients and methods: One hundred twelve consecutive cases of TETs and relative clinico-pathologic fea-tures were collected. Immunohistochemical expression of ALK (clone D5F3) and PD-L1 (clone E1L3N),molecular analysis of EGFR (exons 18–21), c-KIT (exons 9,11,13,14,17), KRAS (exon 2), BRAF (exon 15),PDGFR-alpha (exon 12) and -beta (exons 12, 14, 18), HER-2 (exons 19 and 20) and c-MET (exons 14,17, 18, 19) mutations were performed. Immuno-molecular results were then statistically matched withclinico-pathologic characteristics.Results: Patients were male in 54% of cases, with a median age of 61 years (range 19–83) and affectedmainly by thymoma (78%) in stage II (45%). At molecular analysis, there were 4 c-KIT mutations (occurringin exon 11 V559A, L576P, Y553N and exon 17 D820E) in thymic carcinomas (type C), but not in other tumortypes (p = 0.003). No mutations were detected in other genes and none case was ALK positive. Twenty-nine (26%) cases were PD-L1 positive (65% of thymic carcinomas and 18% of thymomas). High PD-L1expression was statistically associated with WHO classification stage type C (p &lt; 0.001) and Masaoka stageIII–IV disease (p = 0.007). In univariate analysis, WHO classification type C, advanced Masaoka stage andabsence of myasthenia, but not PD-L1 expressions were correlated with worse survival; at multivariateanalysis, only WHO type C confirmed its negative prognostic role.Conclusion: A subset of TETs as thymic carcinomas can harbor c-KIT mutations and elevated PD-L1 expres-sion that could represent targets of potential therapeutic use.

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 - Relationship between Mediterranean diet and asymptomatic peripheral arterial disease in a population of pre-menopausal women [Articolo su rivista]
Mattioli, Anna Vittoria; Coppi, Francesca; Migaldi, Mario; Scicchitano, P.; Ciccone, M. M.; Farinetti, Alberto
abstract

...

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&nbsp;% of the patients were submitted to extended PLND (ePLND). Kaplan–Meier curves confirm the negative prognostic significance of nodal metastases on the overall patients’ survival (PÂ&nbsp;

2016 - Histological and immunohistochemical analysis of meatbased food preparations [Articolo su rivista]
Migaldi, Mario; Giulio, Rossi; Alessandro, Sgambato; Farinetti, Alberto; Mattioli, Anna Vittoria
abstract

This study aimed to verify the possibility to use standard morphologically-based techniques to assess the tissue composition of typical meat-based food preparations. Twenty-six different types of meat-based products were randomly selected and histologically analyzed. A variety of tissues were identified including connective tissue, blood vessels, peripheral nerve, adipose tissue, glandular tissue, skin, bone and cartilage and plant material. Glial fibrillary acidic protein staining was also used to evaluate the presence of brain tissue. The results obtained suggest that most of the analyzed products display a more complex composition than the one suggested by package label. These findings confirms the suitability of standard morphological techniques to examine the histology of meat-based food preparations and suggest their use for the control and quality certification of these types of products

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

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 - Laparoscopic surgery for colorectal cancer is not associated with an increase in the circulating levels of several inflammation-related factors [Articolo su rivista]
Crucitti, A; Corbi, M; Tomaiuolo, Pm; Fanali, C; Mazzari, A; Lucchetti, D; Migaldi, Mario; Sgambato, A.
abstract

It has been hypothesized that inflammatory response triggered by surgery might induce the release of molecules that could promote proliferation, invasion and metastasis of surviving cancer cells. To test this hypothesis, the levels of multiple inflammation-related circulating factors were analyzed in patients undergoing surgery for colorectal cancer. A Luminex xMAP system was used to simultaneously assess levels of IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, FGF, eotaxin, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α and VEGF in 20 colorectal cancer patients and 10 age-matched non-neoplastic patients. In cancer patients analyses were performed at baseline (before surgery) and at different time points (up to 30 days) following laparoscopic surgery. Significantly higher levels of IL-1β, IL-7, IL-8, G-CSF, IFN-γ and TNF-α were detected in colorectal cancer patients compared to controls at baseline. In colorectal cancer patients, circulating levels decreased progressively following surgery and after day 30 post-surgery were no longer different from controls. These findings suggest that expression levels of several cytokines are higher in colorectal cancer patients compared to control subjects and no significant increase in several inflammation-related circulating factors is observed following laparoscopic surgery for cancer. Confirmation and validation in a different and larger cohort of patients are warranted.

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 &lt; 0.05). Conclusion. The detection of histopathological alterations in the mesorectum can give reason of the well-known postoperative complications and long-term sequels.

2014 - Endobronchial metastasis: an epidemiologic and clinicopathologic study of 174 consecutive cases [Articolo su rivista]
Marchioni, Alessandro; Lasagni, Anna; Busca, Annalisa; Cavazza, Alberto; Agostini, Lorenzo; Migaldi, Mario; Corradini, Paolo; Rossi, Giulio
abstract

PURPOSE: Endobronchial metastases from extrapulmonary solid tumors are a rare event and currently available epidemiological and clinico-pathological data mainly derive from anecdotal case reports. METHODS: A series of 174 consecutive cases of endobronchial metastases from extrathoracic solid tumors were collected over a period of 18 years. Immunohistochemistry was performed in 115 cases. Complete imaging features were available in 81 patients, and analysis of the latency period between primitive tumor diagnosis and occurrence of endobronchial metastasis was obtained. RESULTS: Among all bronchoscopic examinations performed in the same period for malignancy, a mean of 5.6 cases per year consisted of endobronchial metastases (range 2-17 cases), with a statistically significant increase when comparing the periods 1992-2000 (65 cases, 37%) and 2001-2009 (109 cases, 63%) (p = 0.05). Overall, 4% of endobronchial biopsies for suspected malignancy disclosed an endobronchial metastasis from extrapulmonary tumor. Breast (52 cases, 30%), colorectal (42 cases, 24%), renal (14%), gastric (6%) and prostate (4.5%) cancers and melanoma (4.5%) were the most common metastatic neoplasms presenting as endobronchial mass. One-hundred fifty-four cases were identified after the primitive tumor diagnosis (metachronous cases, 89%), 11 cases were simultaneously evidenced in extrapulmonary and endobronchial sites (synchronous cases, 6%), while 9 occult metastatic cases (5%) first presented as endobronchial mass (anachronous cases). Overall, mean latency from extrapulmonary tumor diagnosis and endobronchial metastasis was 136 months (range, 1-300 months). The most frequent symptoms were dyspnea (23%), cough (15%) and haemoptysis (12%), while 26% of patients were totally asymptomatic. At radiology, 53% presented as multiple pulmonary nodules, while other cases presented as hilar and mediastinal mass, single peripheral nodule, atelectasis or pleural effusion. CONCLUSIONS: Endobronchial metastases from extrapulmonary tumors account for about 4% of all bronchoscopic biopsies performed for suspected malignancy and in 5% of the cases the metastasis is the first manifestation of the neoplasm.

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 - Large cell carcinoma of the lung: clinically oriented classification integrating immunohistochemistry and molecular biology. [Articolo su rivista]
Rossi, G; Mengoli, Mc; Cavazza, Alessia; Nicoli, D; Barbareschi, M; Cantaloni, C; Papotti, M; Tironi, A; Graziano, P; Paci, Massimiliano; Stefani, Alessandro; Migaldi, Mario; Sartori, Giuliana; Pelosi, G.
abstract

This study aimed at challenging pulmonary large cell carcinoma (LLC) as tumor entity and defining different subgroups according to immunohistochemical and molecular features. Expression of markers specific for glandular (TTF-1, napsin A, cytokeratin 7), squamous cell (p40, p63, cytokeratins 5/6, desmocollin-3), and neuroendocrine (chromogranin, synaptophysin, CD56) differentiation was studied in 121 LCC across their entire histological spectrum also using direct sequencing for epidermal growth factor receptor (EGFR) and v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations and FISH analysis for ALK gene translocation. Survival was not investigated. All 47 large cell neuroendocrine carcinomas demonstrated a true neuroendocrine cell lineage, whereas all 24 basaloid and both 2 lymphoepithelioma-like carcinomas showed squamous cell markers. Eighteen out of 22 clear cell carcinomas had glandular differentiation, with KRAS mutations being present in 39 % of cases, whereas squamous cell differentiation was present in four cases. Eighteen out of 20 large cell carcinomas, not otherwise specified, had glandular differentiation upon immunohistochemistry, with an exon 21 L858R EGFR mutation in one (5 %) tumor, an exon 2 KRAS mutation in eight (40 %) tumors, and an ALK translocation in one (5 %) tumor, whereas two tumors positive for CK7 and CK5/6 and negative for all other markers were considered adenocarcinoma. All six LCC of rhabdoid type expressed TTF-1 and/or CK7, three of which also harbored KRAS mutations. When positive and negative immunohistochemical staining for these markers was combined, three subsets of LCC emerged exhibiting glandular, squamous, and neuroendocrine differentiation. Molecular alterations were restricted to tumors classified as adenocarcinoma. Stratifying LCC into specific categories using immunohistochemistry and molecular analysis may significantly impact on the choice of therapy.

2014 - Lung cancer histologic and immunohistochemical heterogeneity in the era of molecular therapies: analysis of 172 consecutive surgically resected, entirely sampled pulmonary carcinomas [Articolo su rivista]
Cadioli, Annamaria; Rossi, Giulio; Costantini, Matteo; Cavazza, Alberto; Migaldi, Mario; Colby, Thomas V.
abstract

On the basis of seminal studies in the 1980s, appreciable histologic heterogeneity, ranging from 45% to 70% of cases, may be encountered in lung cancer. However, the epidemiologic and histologic landscape of lung cancer in the last 3 decades has radically changed. In this study, 172 consecutive surgically resected primary lung carcinomas evaluated from 2010 to 2012 were entirely sampled and examined according to current histologic classifications. In 129 cases, a positive preoperative biopsy was also available. Major histologic heterogeneity (a single tumor showing at least 2 different histologic types) and minor histologic heterogeneity (a single tumor showing just 1 histologic type but at least 2 different growth patterns) were evaluated in all cases. Immunohistochemical heterogeneity (ie, "aberrant" staining) was also assessed using a panel of markers of adenocarcinoma (TTF-1, napsin, and CK7), squamous cell carcinoma (p63, CK5/6), and neuroendocrine differentiation (chromogranin and synaptophysin), both on positive biopsies and surgical specimens. Overall, major and minor histologic heterogeneity on resections were disclosed in 4% (7 cases) and 50.6% (87 cases), respectively, whereas just 1 case of minor heterogeneity (pleomorphic carcinoma) was observed on biopsies. Minor heterogeneity was limited to adenocarcinomas (82.6%, 81/98 cases) and sarcomatoid carcinomas (6 pleomorphic types among 8 sarcomatoid carcinomas). Immunohistochemical heterogeneity was recorded in 22.6% of the cases, with expression of p63 and CK5/6 in a subset of adenocarcinomas (25 cases, 25.5%), CK7 in 17.4% of squamous cell carcinomas, and synaptophysin in 6 cases of non-neuroendocrine tumors (4%, 6/155). The high rate of adenocarcinomas, accounting for 57% (98 cases) of 172 consecutively resected lung cancers, reflects the new scenario of thoracic oncology and may explain the significant lower rate of major histologic heterogeneity (4%) and the higher frequency of different architectural patterns (minor heterogeneity) that we found in lung cancer compared with previous studies.

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 - Serratia marcescens in a neonatal intensive care unit: two long-term multiclone outbreaks in a 10-year observational study [Articolo su rivista]
Casolari, Chiara; Pecorari, Monica; Della Casa, Elisa; Cattani, Silvia; Venturelli, Claudia; Fabio, Giuliana; Tagliazucchi, Sara; Serpini, Giulia Fregni; Migaldi, Mario; Marchegiano, Patrizia; Rumpianesi, Fabio; Ferrari, Fabrizio
abstract

We investigated two consecutive Serratia marcescens (S. marcescens) outbreaks which occurred in a neonatal intensive care unit (NICU) of a tertiary level hospital in North Italy in a period of 10 years (January 2003-December 2012). Risk factors associated with S. marcescens acquisition were evaluated by a retrospective case-control study. A total of 21,011 clinical samples was examined: S. marcescens occurred in 127 neonates: 43 developed infection and 3 died. Seven clusters were recorded due to 12 unrelated clones which persisted for years in the ward, although no environmental source was found. The main epidemic clone A sustaining the first cluster in 2003 reappeared in 2010 as an extended spectrum ?-lactamase (ESBL)-producing strain and supporting the second epidemic. Birth weight, gestational age, use of invasive devices and length of stay in the ward were significantly related to S. marcescens acquisition. The opening of a new ward for non-intensive care-requiring neonates, strict adherence to alcoholic hand disinfection, the timely identification and isolation of infected and colonized neonates assisted in containing the epidemics. Genotyping was effective in tracing the evolution and dynamics of the clones demonstrating their long-term persistence in the ward.

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&lt;0.001 and p&lt;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 role of molecular analyses in the diagnosis and treatment of non-small-cell lung carcinomas [Articolo su rivista]
Rossi, Giulio; Graziano, Paolo; Leone, Alvaro; Migaldi, Mario; Califano, Raffaele
abstract

Non-small-cell lung cancer (NSCLC) subtyping has recently been a key factor in determining patient management with novel drugs. In addition, the identification of distinct oncogenic driver mutations frequently associated with NSCLC histotype and coupled to the clinical responses to targeted therapies have revolutionized the impact of histologic type and molecular biomarkers in lung cancer. Several molecular alterations involving different genes (EGFR, KRAS, ALK, BRAF, and HER2) seem to have a remarkable predilection for adenocarcinoma and specific inhibitors of EGFR and ALK are now available for patients with adenocarcinoma harboring the relevant gene alterations. The efficacy of histology-based and molecular-targeted therapies had a deep impact in (1) re-defining classification of lung cancer (particularly adenocarcinomas) and (2) routine clinical practice of pathologists involved in optimization of handling of tissue samples in order to guarantee NSCLC subtyping with the help of immunohistochemistry and adequately preserve tumor cells for molecular analysis. In agreement with the modern multidisciplinary approach to lung cancer, we reviewed here the diagnostic and predictive value of molecular biomarkers according to the clinical, pathologic, and molecular biologist viewpoints.

2013 - The usefulness of Moynihan questionnaire in the evaluation of knowledge on healthy diet of patients undergoing cardiology rehabilitation [Articolo su rivista]
Rosi, C; Pennella, Sonia; Fantuzzi, Anna Laura; Pedrazzi, P; Passalacqua, M; Gavioli, M; Migaldi, Mario; Farinetti, Alberto; Mattioli, Anna Vittoria
abstract

BACKGROUND: The aim of study was to test the usefulness of Moynihan questionnaire in the evaluation of knowledge on healthy diet of patients undergoing cardiology rehabilitation. METHODS: We enrolled 51 patients (pts): 41 men and 10 women, mean age 67.97 +/-11.2 years. The case study included: 21 pts that underwent coronary bypass surgery, 16 pts replaced plastic tube, 14 pts had surgery for the other reasons. All pts underwent nutritional investigation by a dietitian. Anthropometric and biochemical parameters were detected and, by the end, the Moynihan questionnaire was administrated. Pts underwent nutritional coaching, and questionnaire and dietary assessment were rechecked after 3 months. RESULTS: At baseline, the mean Questionnaire score was 22.4 +/- 3.2 points, decreased to 20.6 +/-3.1 points after 3 months (p&lt;0.05). A detailed analysis of the questions showed that the major informations gaps were related to consumption of fruits and vegetables, consumption of fat and salt. In addition pts have acquired more general knowledge about food composition. CONCLUSIONS: The Moynihan questionnaire is an useful instrument of evaluation of dietary knowledge even in selected patients population. In the present study involving patients after cardiac surgery the main difficulties were related to high age of pts, the low cultural level and, mainly, to the post-surgery stress. However, an increase of correct answers as well as an increased knowledge about food composition were detected after educational intervention performed by the dietitian.

2012 - Activating c-KIT mutations in a subset of thymic carcinoma and response to different c-KIT inhibitors. [Articolo su rivista]
Schirosi, L; Nannini, N; Nicoli, D; Cavazza, A; Valli, R; Buti, S; Garagnani, Lorella; Sartori, G; Calabrese, F; Marchetti, A; Buttitta, F; Felicioni, L; Migaldi, Mario; Rea, F; Di Chiara, F; Mengoli, Mc; Rossi, G.
abstract

To analyze a multi-institutional series of type C thymic carcinomas (TCs) (including neuroendocrine tumors), focusing on the expression and mutations of c-KIT. Immunohistochemical expression of c-KIT/CD117, p63, CD5 and neuroendocrine markers, as well as mutational analysis of c-KIT exons 9, 11, 13, 14, 17 by direct sequencing of 48 cases of TCs. Immunohistochemical and molecular data were statistically crossed with clinicopathological features. Overall, 29 tumors (60%) expressed CD117, 69% were positive for CD5 and 85% (41 cases) for p63. Neuroendocrine markers stained all six atypical carcinoids and five poorly-differentiated thymic squamous cell carcinomas. Overall, six CD117-positive cases (12.5%) showed c-KIT mutation. No mutation was detected in CD117-negative tumors and carcinoids. All the mutations were found in poorly-differentiated thymic squamous cell carcinomas expressing CD117, CD5, p63 and lacking neuroendocrine markers (6 of 12 cases with these features). Mutations involved exon 11 (four cases: V559A, L576P, Y553N, W557R), exon 9 (E490K) and exon 17 (D820E).Conclusions: All TCs need an immunohistochemical screening with CD117, while c-KIT mutation analysis is mandatory only in CD117-positive cases, particularly when coexpressing CD5 and p63, lacking neuroendocrine differentiation. The finding of c-KIT mutation can predict efficacy with different c-KIT inhibitors.

2012 - Adherence to guidelines in Aspirin Administration in Real-Life Patients with Low Cardiovascular risk [Abstract in Rivista]
Pennella, Sonia; Farinetti, Alberto; Migaldi, Mario; Mattioli, Anna Vittoria
abstract

adherence to aspirin

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 &lt; 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 - Low prevalence of human papillomavirus infection in the healthy oral mucosa of a Northern Italian population. [Articolo su rivista]
Migaldi, Mario; Pecorari, M; Forbicini, G; Nanni, N; Grottola, Antonella; Grandi, T; Delle Donne, G; Leocata, P; Trovato, D; Sgambato, A.
abstract

Oral cancer is the sixth most common malignancy in developed countries, representing almost 3% of malignant tumors. Tobacco use and alcohol consumption are well-established risk factors. However, the observation that most patients with oral cancer have not been exposed to these risk factors suggests that additional causes may promote oral carcinogenesis. A link has been suggested between human papillomavirus (HPV) and oral cavity cancer but the significance of HPV contribution to oral carcinogenesis as well as the prevalence of HPV infection in normal oral cavity mucosa remains debated. Methods:  In this study, the prevalence of oral HPV infection was evaluated in 81 randomly selected Northern Italian subjects with clinically normal oral mucosa using a nested PCR on DNA extracted by oral smears. Results and conclusions:  No HPV-related lesions were detectable in any of the smears analyzed by cytological approach. nPCR identified HPV DNA in only one (1.2%) of the specimens obtained from clinically healthy oral mucosa and subsequent characterization assigned the positive case to HPV type 90. These data suggest that the incidence of HPV infection in the healthy population might be very low and that other risk factors are likely responsible to promote oral carcinogenesis.

2012 - MUC5AC, cytokeratin 20 and HER2 expression and K-RAS mutations within mucinogenic growth in congenital pulmonary airway malformations [Articolo su rivista]
Rossi, G; Gasser, B; Sartori, G; Migaldi, Mario; Costantini, M; Mengoli, Mc; Piccioli, S; Cavazza, A; Rivasi, Francesco
abstract

Histopathology MUC5AC, cytokeratin 20 and HER2 expression and K-RAS mutations within mucinogenic growth in congenital pulmonary airway malformations Aims:  To analyse the expression of several mucins (MUC1, MUC2, MUC3, MUC5AC and MUC6), epidermal growth factor receptor (EGFR), v-erb-b2 erythroblastic leukaemia viral oncogene homologue 2 (HER2), thyroid transcription factor-1 (TTF-1), caudal type homeobox 2 (CDX2) and cytokeratin 20 (CK20), and the presence of mutations of EGFR, KRAS and HER2 in congenital pulmonary airway malformations (CPAM). Methods and results:  Forty-one cases of CPAM and six pulmonary sequestrations were included. TTF-1 expression was observed in all cases but was not seen in mucinogenic growths in CPAM. CDX2 expression was completely negative. MUC1 expression was noted in 12 (29%) CPAM and in 33% sequestrations. MUC5AC was noted in only five cases (26%) by immunohistochemistry and was found in the mucinogenic proliferations of type 1 CPAM. No immunolabelling was noted for the other mucins. EGFR was expressed variably in almost all cases, while HER2 and CK20 was seen exclusively in the mucinogenic proliferations. All mucinous growths were characterized by KRAS mutations. No EGFR and HER2 gene alterations were identified. Conclusions:  KRAS mutations and MUC5AC, CK20 and HER2 expression was seen in all mucinogenic proliferations, supporting the neoplastic nature of these cytologically bland growths. These findings emphasize the importance of complete surgical resection of such lesions.

2012 - Prognostic significance of MGMT gene promoter methylation in differently treated metastatic melanomas [Articolo su rivista]
A. M., Cesinaro; Sartori, Giuliana; Migaldi, Mario; L., Schirosi; Pellacani, Giovanni; G., Collina; Maiorana, Antonino
abstract

Aims: The methylation status of the MGMT gene promoter,considered of prognostic significance by enhancing chemosensitivityto alkylating drugs in gliomas and melanomas, wasevaluated in a series of primary melanomas and metastasesof patients treated with different therapies, to identify anycorrelation with the patients’ outcome or response to differenttherapeutic regimens.Methods: Twenty-nine primary melanomas and 74 metastases,collected from 52 patients, were assessed forMGMT gene promoter methylation using a standardmethylation specific PCR-based method. All materials wereformalin fixed and paraffin embedded.Results: One of 29 primary melanomas (3.4%) and 22 of74 metastases (29.7%) showed MGMT gene promotermethylation. MGMT methylation was more frequent invisceral (17/40, 42.5%) than in cutaneous/lymph nodemetastases (5/34, 14.7%) ( p¼0.019). Both disease free(DFS) and overall survival (OS) were significantly longerin patients with methylated metastases ( p¼0.009 andp¼0.007, respectively). No correlations were found amongmethylation, therapeutic regimens and DFS or OS.Conclusions: MGMTmethylation appears to be a late event inthe biological history of melanoma and is more frequentlyseen in visceral metastases. The MGMT gene promotermethylation in metastatic disease is associated with longersurvival, irrespective of therapy. Thus it could be considereda prognostic factor in metastatic melanoma.

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 - 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 - INI1 immunohistochemical expression in glioblastoma: correlation with MGMT gene promoter methylation status and patient survival [Articolo su rivista]
E., Zunarelli; N., Bigiani; Sartori, Giuliana; Migaldi, Mario; A., Sgambato; Maiorana, Antonino
abstract

Aims: INI1 expression and its correlation with MGMT gene promoter methylation status and follow-up was investigated in 77 surgically removed glioblastomas then treated with radiotherapy (RT) or RT plus temozolomide (TMZ).Methods: INI1 was determined by immunohistochemistry and MGMT by methylation-specific PCR.Results: INI1 was expressed in 83.1% of cases. The median overall survival (OS) was 13.6 months in INI1+ tumours and 7.2 months in INI1- tumours. 31.3% of patients with INI1+ tumours were alive compared with 15.4% of patients with INI1- tumours. MGMT methylation was detected in 31.2% of cases. OS was significantly different between patients with methylated tumours and un-methylated tumours (p < 0.04), and between patients with RT+ TMZ and RT alone (p < 0.001). Considering both treatment and MGMT, the difference in OS was significant (p < 0.002). The difference in OS according to MGMT and INI1 was significant (p < 0.04). The longest median OS was recorded among methylated and INI1+ tumours. Among un-methylated tumours, the median OS was 11.1 months in INI1+ and 6.5 months in INI1- tumours. No patients were alive with un-methylated and INI1- tumours.Conclusions: Loss of INI1 immunohistochemical expression in glioblastoma may be indicating an underlying molecular aberration accounting for the more aggressive clinical behaviour.

2011 - Urotensin II receptor predicts the clinical outcome of prostate cancer patients and is involved in the regulation of motility of prostate adenocarcinoma cells. [Articolo su rivista]
Grieco, P; Franco, R; Bozzuto, G; Toccacieli, L; Sgambato, A; Marra, M; Zappavigna, S; Migaldi, Mario; Rossi, G; Striano, S; Marra, L; Gallo, L; Cittadini, A; Botti, G; Novellino, E; Molinari, A; Budillon, A; Caraglia, M.
abstract

Urotensin II (UT-II) is a potent vasoconstrictor peptide and its receptor (UTR) was correlated with human cortico-adrenal carcinoma proliferation. In this study, we have evaluated the correlation between UTR expression and prognosis of human prostate adenocarcinoma and the involvement of this receptor in the regulation of biological properties on both in vivo and in vitro models. UTR mRNA and protein, evaluated by real-time PCR and Western blotting, respectively, were expressed at high levels only in androgen-dependent LNCaP cells. In order to investigate UTR changes occurring in human prostate tumorigenesis, we have also evaluated the expression of UTR in vivo in 195 human prostate tissue samples. UTR was always expressed at low intensity in hyperplastic tissues and at high intensity in well-differentiated carcinomas (Gleason 2-3). Moreover, we have evaluated the effects of an antagonist of UTR, urantide on migration and invasion of LNCaP cells. Urantide induced a dose-dependent decrease of motility and invasion of LNCaP cells whose characteristic ameboid movement seems to be advantageous for their malignancy. These effects were paralleled by down-regulating the autophosphorylation of focal adhesion kinase and the integrin surface expression on LNCaP cells. The effects on cell motility and invasion were likely due to the inhibition of RhoA activity induced by both urantide and shRNA UTR. These data suggest that UTR can be considered a prognostic marker in human prostate adenocarcinoma patients.

2010 - Alterations of 9p21 analysed by FISH and MLPA distinguish atypical spitzoid melanocytic tumours from conventional Spitz's nevi but do not predict their biological behaviour. [Articolo su rivista]
Cesinaro, Am; Schirosi, L; Bettelli, S; Migaldi, Mario; Maiorana, Antonino
abstract

Alterations of 9p21 analysed by FISH and MLPA distinguish atypical spitzoid melanocytic tumours from conventional Spitz's nevi but do not predict their biological behaviour Aim:  Histopathological criteria alone cannot predict the biological behaviour of spitzoid melanocytic tumours. The aim of this study was to investigate whether 9p21 status influence the prognosis of the spitzoid melanocytic tumours, peculiar lesions whose biological behaviour cannot be predicted by histopathological criteria alone. Methods and results:  Twenty-eight atypical spitzoid tumours, 12 conventional Spitz's nevi and one congenital Spitz's nevus were studied by fluorescent in-situ hybridization (FISH) and multiple ligation-dependent probe amplification (MLPA) for the presence of 9p21 deletion. The 28 patients were aged 3-56 years (mean 32, median 35), and follow-up ranged between 4 and 156 months (mean 51, median 48). Eight patients (28.5%) experienced lymph node metastasis (three cases with macrometastasis and five with micrometastasis). Of those with macrometastasis, two are alive after 159 and 26 months, whereas a third developed widespread metastases and died after 26 months. All of the other patients are alive. Statistically, the thickness (P = 0.01) and the diameter (P = 0.009) of the lesions significantly correlated with metastasis. Deletion of 9p21 by FISH analysis was observed in eight spitzoid tumours (28.5%), and MLPA demonstrated alterations of 9p21, particularly deletion of CDKN2A, in the same lesions, whereas all Spitz's nevi, except the congenital one, were of unaltered 9p21 status (P < 0.0001). Deletion of 9p21/CDKN2A did not correlate with the presence of metastasis. Conclusion:  Alterations at 9p21 locus are significantly more frequent in spitzoid tumours than in Spitz's nevi, but do not predict their biological behaviour.

2010 - Expression of alpha-dystroglycan correlates with tumour grade and predicts survival in oral squamous cell carcinoma. [Articolo su rivista]
Sgambato, A; Caredda, E; Leocata, P; Rossi, G; Boninsegna, A; Vitale, A; Grandi, T; Cittadini, A; Migaldi, Mario
abstract

Dystroglycan (DG) is a non-integrin adhesion molecule connecting the extracellular matrix to the actin cytoskeleton. Decreased expression of DG has been reported in several human cancers and related to tumour aggressiveness. Expression of the alpha-DG subunit was evaluated by immunostaining in a series of oral squamous cell carcinoma (OSCC) and its relation with traditional prognostic indicators and with the clinical outcome of the patients was evaluated. Alpha-DG expression was easily detected in normal epithelium with a mean percentage of positive cells >80% but was undetectable in a significant fraction (59%) of OSCC. Loss of alpha-DG staining correlated with higher tumour grade (p = 0.04) and stage (p = 0.01), with nodal involvement (p = 0.001) and with an increased risk of recurrence (p = 0.002) and death (p = 0.004) in a univariate analysis, but it was not confirmed as an independent predictor of clinical outcome in a multivariate analysis. Loss of alpha-DG expression, which corresponds to loss of a functional DG complex, is a frequent event in human OSCC. Further studies are warranted on the role of this molecule in the entire multistep process of oral squamous tumorigenesis.

2010 - Loss of nuclear p27(kip1) and alpha-dystroglycan is a frequent event and is a strong predictor of poor outcome in renal cell carcinoma. [Articolo su rivista]
Sgambato, A; Camerini, A; Genovese, G; De Luca, F; Viacava, P; Migaldi, Mario; Boninsegna, A; Cecchi, M; Sepich, Ca; Rossi, G; Arena, V; Cittadini, A; Amoroso, D.
abstract

Expression levels of p27(kip1), a negative regulator of the G1 phase of the cell cycle, and 8-hydroxydeoxyguanosine (8-OHdG), a marker of oxidative DNA damage, were assessed by immunostaining in a series of renal cell carcinomas (RCCs) and their prognostic significance was evaluated. Expression of p27(kip1) as well as of the alpha-subunit of the dystroglycan (DG) complex, previously reported to be altered in RCC, was also evaluated by western blot analysis. Nuclear expression of p27(kip1) was reduced in a significant fraction of tumors and low p27(kip1) staining correlated with higher tumor grade (P < 0.01). Recurrence and death from clear cell RCCs were significantly more frequent in p27(kip1)-low expressing tumors and Kaplan-Meier curves showed a significant separation between high vs low expressor groups for both disease-free (P = 0.011) and overall (P = 0.002) survival. Low nuclear expression of p27(kip1) as well as loss of alpha-DG were confirmed to be independent prognostic parameters at a multivariate analysis and the simultaneous loss of both molecules defined a "high-risk" group of patients with increased risk of recurrence (RR = 28.7; P = 0.01) and death (RR = 12.9; P = 0.03). No significant correlation with clinical or pathological parameters was found for 8-OHdG staining. Western blot analyses suggested a post-translational mechanism for the loss of alpha-DG expression and demonstrated that cytoplasmic dislocation of the protein contributes to the loss of active nuclear p27(kip1). Loss of nuclear p27(kip1) is a frequent event in human RCCs and is a powerful predictor of poor outcome which, in combination with low DG expression, could help to identify high-risk patients with clear cell RCC.

2009 - Cyclin E correlates with manganese superoxide dismutase expression and predicts survival in early breast cancer patients receiving adjuvant epirubicin-based chemotherapy [Articolo su rivista]
Sgambato, A; Camerini, A; Collecchi, P; Graziani, C; Bevilacqua, G; Capodanno, A; Migaldi, Mario; Masciullo, V; Scambia, G; Rossi, G; Cittadini, A; Amoroso, D.
abstract

Anthracycline-based chemotherapy represents a milestone in the treatment of breast cancer. We previously demonstrated in an in vitro model that cyclin E overexpression is associated with increased expression of manganese superoxide dismutase (MnSOD) and resistance to doxorubicin. In the present study, immunohistochemical expression of cyclin E and MnSOD was evaluated in 134 early breast cancer patients receiving adjuvant epirubicin-based chemotherapy regimens containing epirubicin. Both parameters were correlated with the available clinicopathological parameters and with the outcome of patients. Overexpression of cyclin E and MnSOD was detected in 46 (34.3%) and 56 (41.8%) patients, respectively, and expression levels of the two proteins were related. Disease-free and alive patients displayed a lower mean percentage of cyclin E-expressing cells than relapsed and dead patients, respectively. Kaplan–Meier survival analysis demonstrated a significant separation between high versus low cyclin E-expressing tumors in terms of overall survival (P = 0.038 by log-rank). Similar results were obtained considering the subset of node-negative patients separately. No significant relationship with patient outcome was observed for MnSOD expression levels. At multivariate analysis cyclin E failed to demonstrate an independent prognostic value. In conclusion, the results of the present study support previous evidence that increased cyclin E expression is associated with higher MnSOD expression levels and poorer outcome, at least as evaluated in terms of overall survival. Further studies are warranted to evaluate the usefulness of cyclin E as a prognostic marker to identify breast cancer patients at higher risk of death from the disease when treated with adjuvant anthracycline-based therapy.

2009 - EGFR and K-ras mutations along the spectrum of pulmonary epithelial tumors of the lung and elaboration of a combined clinicopathologic and molecular scoring system to predict clinical responsiveness to egfr inhibitors [Articolo su rivista]
Sartori, G; Cavazza, A; Sgambato, A; Marchioni, A; Barbieri, F; Longo, L; Bavieri, M; Murer, B; Meschiari, E; Tamberi, S; Cadioli, A; Luppi, F; Migaldi, Mario; Rossi, G.
abstract

We tested 418 neoplasms along the whole spectrum of primary lung tumor histotypes for epidermal growth factor receptor (EGFR) and K-ras mutations. Clinicopathologic data from 154 patients undergoing treatment with EGFR tyrosine kinase inhibitors (TKIs) were retrospectively studied. A scoring system assigning a score for each positive or negative characteristic (+1, female sex, nonsmoking status, adenocarcinoma histotype, Asian ethnicity, and EGFR mutation; −1, current smoker and K-ras mutation; and 0, male sex, ex-smoker, nonadenocarcinoma histotype, and no mutations) was elaborated and tested with EGFR-TKI response.Salivary gland–type, mucin-rich, and neuroendocrine tumors do not harbor EGFR mutations. A subset of nonmucinous adenocarcinomas, not necessarily of the bronchioloalveolar type, is related to EGFR mutations. Three probability groups significantly correlating with response to EGFR-TKIs were identified. Of note, the addition of molecular results did not significantly change the predictive value obtained by the combination of clinicopathologic characteristics alone in this scoring system. K-ras mutations, significantly associated with the mucin-secreting type of adenocarcinoma, consistently predict lack of response in white patients.

2009 - Psoriasis vs allergic contact dermatitis in palms and soles: A quantitative histologic and immunohistochemical study [Articolo su rivista]
Cesinaro, Am; Nannini, N; Migaldi, Mario; Pepe, P; Maiorana, Antonino
abstract

A systematic histologic and immunohistochemical study of cases of psoriasis (PSO) and allergic contact dermatitis (ACD) in palmo-plantar skin was performed to find differences between these two diseases that usually show overlapping features in these specific sites. Skin biopsies from 42 (22 female, 20 male) patients were evaluated for several histopathologic parameters and immunohistochemistry was applied to quantify keratinocytic proliferation, the number of dendritic cells (DCs) and the phenotype of the mononuclear cell infiltrate. Regular epidermal hyperplasia and marked parakeratosis were found to be more frequent in PSO than in ACD cases, but only the first parameter reached the level of significance (p = 0.03). The number of S100 protein-positive DCs was significantly higher in ACD (p = 0.006), whereas keratinocytic proliferation, studied with Mib-1, was found to be higher in PSO than in ACD, but the difference was not statistically significant. No significant difference was detected in the number of CD4+, CD8+ and bcl2+ lymphocytes in PSO and ACD cases. In the palms and soles, the finding of irregular epidermal hyperplasia and the detection of a higher number of S100 protein-positive DCs favor the diagnosis of ACD over PSO. The differential diagnosis between PSO and ACD can be practically approached using a histopathologic parameter and a commercially available antibody.

2008 - A subset of lung adenocarcinomas and atypical adenomatous hyperplasia-associated foci are genotypically related: an EGFR, HER2, and K-ras mutational analysis. [Articolo su rivista]
Sartori, G; Cavazza, A; Bertolini, F; Longo, L; Marchioni, A; Costantini, M; Barbieri, F; Migaldi, Mario; Rossi, G.
abstract

Atypical adenomatous hyperplasia (AAH) is considered the preinvasive lesion of pulmonary adenocarcinoma, and mutations of EGFR, HER2, and K-ras are involved in the early stage of lung adenocarcinoma carcinogenesis, also predicting clinical response to anti-EGFR small molecule inhibitors. We analyzed 18 cases of primary lung adenocarcinoma with concomitant AAH foci from 13 patients for mutations of EGFR (exons 18-21), HER2 (exons 19-20), and K-ras (exon 2) by direct sequencing polymerase chain reaction. Among mutated cases, concordant mutations of EGFR or K-ras in adenocarcinoma and related AAH were observed in 5 (63%) of 8 cases. In particular, 3 of 4 adenocarcinomas with EGFR mutations (all L858R point mutations in women, never or former smokers) had a concomitant and identical mutation in AAH, and 2 of 4 adenocarcinomas with K-ras mutations (both at codon 12 in women, a never and a current smoker) showed the same mutation in concomitant AAH. All cases were wild-type for HER2. Mutations of EGFR and K-ras genes represent an early event in lung adenocarcinomagenesis, and AAH convincingly seems to be a precursor lesion in a subset of cases of adenocarcinoma.

2008 - Pleuro-pulmonary solitary fibrous tumors: A clinicopathologic, immunohistochemical, and molecular study of 88 cases confirming the prognostic value of de perrot staging system and p53 expression, and evaluating the role of c-kit, BRAF, PDGFRs (α/β), c-met, and EGFR [Articolo su rivista]
Schirosi, L; Lantuejoul, S; Cavazza, A; Murer, B; Yves Brichon, P; Migaldi, Mario; Sartori, G; Sgambato, A; Rossi, G.
abstract

Pleuro-pulmonary solitary fibrous tumor (SFT) is a relatively uncommon mesenchymal neoplasm of uncertain histogenesis, unknown molecular features, and unpredictable clinical behavior. Although complete resection is universally accepted as the most important single prognostic factor, some clinicopathologic characteristics (gross appearance, tumor size, mitotic index, tumor necrosis, hypercellularity, and pleomorphism) are related to patient outcome, and a staging system based on these parameters with practical therapeutical implications has been recently proposed by de Perrot et al. Here, 88 pleuro-pulmonary SFTs were collected and clinicopathologic characteristics including de Perrot classification, patients' follow-up, p53 expression, and several "targetable" kinases [c-kit, v-raf murine sarcoma viral oncogene homolog B1, platelet-derived growth factor receptor (PDGFR)-alpha/beta, c-met, epidermal growth factor receptor (EGFR)] were retrospectively analyzed. Fifty-two cases (59%) had at least 1 clinicopathologic feature related to malignancy, whereas mortality and recurrences occurred in 10.2% and 18.2% of the cases, respectively. de Perrot staging and high p53 expression were significantly related to the conventional clinicopathologic prognostic features as well as to overall survival (OS) and disease-free survival (DFS) (P&lt;0.001). At multivariate analysis, high p53 expression and tumor necrosis were the only parameters significantly associated with OS and DFS (P=0.017 and P=0.012, respectively). Immunohistochemical expression was frequently detected for PDGFR-alpha (97.7%), PDGFR-beta (86.5%), and hepatocyte growth factor receptor (96.6%), whereas missense mutations were only identified in 2 cases both involving PDGFR-beta (exons 18 and 20). We conclude that de Perrot stratification of SFT is a reliable prognostic indicator and merits consideration in view of its suggestions for the management of these tumors in daily practice. p53 expression may represent a valid and easy-to-test prognostic factor significantly related to OS and DFS. Although mutations of the corresponding genes are rare events in SFT, PDGFR-alpha/beta, and hepatocyte growth factor receptor tyrosine kinases should be further investigated given the availability of specific inhibitory molecules which might provide useful and novel therapeutical approaches for SFT patients.

2008 - Thymoma classification: does it matter? [Articolo su rivista]
Rossi, G; Costantini, M; Tagliavini, E; Barbieri, F; Migaldi, Mario; Casali, C.
abstract

We read with interest the Commentary on the classification of thymomas by Addis and den Bakker in the May (2008) issue of Histopathology, and we would add some further information on the controversial prognostic value of histological classification based on our recent experience with these intriguing neoplasms.

2008 - Tumor cell proliferation and microsatellite alterations in human ameloblastoma [Articolo su rivista]
Migaldi, Mario; Sartori, G; Rossi, G; Cittadini, A; Sgambato, A.
abstract

Ameloblastoma is the most common odontogenic tumor. It can exhibit a variety of histological patterns, a great infiltrative potential and a high recurrence rate. Mutations in microsatellite sequences are a hallmark of neoplastic transformation but little is known about their role in ameloblastoma development. In this study DNA was extracted from laser-microdissected samples of 24 ameloblastomas and was analyzed for the status of 22 microsatellite loci. The occurrence and the pattern of microsatellite alterations, in form of loss or length variation, was evaluated and correlated with the Ki67 labeling index and with other clinicopathologic parameters. The prognostic significance of these alterations was also evaluated. High Ki67 expression was significantly associated with a shorter disease-free survival (p=0.003 by log-rank test). Alterations of at least one of the selected loci was observed in all (100%) the ameloblastomas analyzed with a mean of 4 altered microsatellites for each tumor. The microsatellites most frequently altered were D9S747 and D11S488 (42%). All the other loci analyzed were altered in less than 40% of cases and some of them (D3S1312, D3S1300, IFNA, D9S164, D13S176 and TP53) did not show alterations in any of the ameloblastomas analyzed. No relationship was observed between the occurrence of microsatellite alterations and other parameters, such as patients age and gender, tumor size, localization and histotype. The occurrence of microsatellite alterations was more frequent in tumors displaying a high Ki67 labeling index (p=0.03) and in a univariate analysis was predictor of an increased risk of disease recurrence (p=0.039 by log-rank test). These findings demonstrate that microsatellite alterations are frequent event in ameloblastomas. They also suggest that evaluation of tumor cells proliferative activity and microsatellite alterations may be helpful to stratify ameloblastomas prognostically and to predict the clinical behavior of these tumors.

2007 - Dystroglycan expression is reduced during prostate tumorigenesis and is regulated by androgens in prostate cancer cells. [Articolo su rivista]
A., Sgambato; B., DE PAOLA; Migaldi, Mario; M., DI SALVATORE; A., Rettino; G., Rossi; B., Faraglia; A., Boninsegna; Maiorana, Antonino; A., Cittadini
abstract

Prostate cancer, the most frequently diagnosed cancer in Western men, can display a high variability in term of clinical aggressiveness and prognosis and none of the available markers is able to accurately predict its clinical course. Dystroglycan (DG), a non-integrin adhesion molecule, is a complex formed by two subunits, alpha- and beta-DG, which bind to extracellular matrix molecules and cytoskeleton, respectively. DG expression is frequently reduced in human cancers and has been related to tumor grade and aggressiveness. This study investigated the role of DG in human prostate tumorigenesis and its suitability as a prognostic marker. The expression level of extracellular alpha-DG subunit was frequently reduced in human prostate cancer cell lines and primary tumors and the percentage of positive tumor cells was significantly further decreased in vivo following androgen ablation therapy (median = 1%) compared to pre-treatment samples (median = 28%). A significant relationship was observed between alpha-DG staining on the post-treatment samples and tumor recurrence. A dose- and time-dependent decrease of DG expression also occurred in human prostate cancer cells following treatment with the anti-androgen flutamide. Stable expression of an exogenous DG cDNA in the LNCaP human prostate carcinoma cell line resulted in a marked inhibition of both anchorage-dependent and independent growth and of the in vivo tumorigenicity. These findings confirm and extend previous evidence that disturbances in the function of the DG complex might contribute to the definition of the malignant behavior of prostate cancer cells and suggest that androgens might regulate DG expression in these cells.

2007 - Expression of dystroglycan correlates with tumor grade and predicts survival in renal cell carcinoma [Articolo su rivista]
Sgambato, A; Camerini, A; Amoroso, D; Genovese, G; De Luca, F; Cecchi, M; Migaldi, Mario; Rettino, A; Valsuani, C; Tartarelli, G; Donati, S; Siclari, O; Rossi, G; Cittadini, A.
abstract

The dystroglycan (DG) complex is a transmembrane glycoprotein that forms a continuous link from the extracellular matrix to the actin cytoskeleton. Deregulated expression of DG has been reported in a variety of human malignancies and related to tumor aggressiveness. In this study expression of the DG subunit was evaluated by immunostaining in a series of renal epithelial cancers and its relation with traditional prognostic indicators and with the clinical outcome of the patients was evaluated. alphaDG expression was undetectable in a significant fraction of tumors (54%). In renal cell carcinomas (RCC) loss of alpha-DG staining correlated with higher tumor grade (p = 0.02) but not with tumor stage nor tumor size. In clear cell RCC patients loss of alphaDG staining correlated with an increased risk of recurrence (p = 0.002 by log-rank test) and death (p = 0.004) also when patients with lower grade or stage tumors were analyzed separately. In a multivariate analysis loss of DG staining confirmed to be and independent predictor of shorter disease-free (p = 0.001; RR = 4.9) and overall (p = 0.009; RR = 4.9) survival stronger than tumor grade and size. These findings demonstrate that loss of alphaDG expression, which correspond to loss of a functional DG complex, is a frequent event in human renal tumorigenesis and is an independent predictor of early recurrence and death for patients with clear cell RCC.

2007 - Unusual concurrent detection by polymerase chain reaction of Bartonella henselae and parvovirus b19 in an immunocompetent child with erythema nodosum and hepatic granulomatous disease [Articolo su rivista]
Casolari, Chiara; M., Pecorari; W., Gennari; N., Nanni; Migaldi, Mario; N., Guaraldi; S., Tagliazucchi; Bertoli, G.; Portolani, Marinella; Balli, Fiorella; Fabio, Giuliana; Sabbatini, Anna Maria Teresa; Alu', Milena; F., Rumpianesi
abstract

We report an unusual case of documented Bartonella henselae genotype I from hepatic tissue in an Italian immunocompetent girl presenting with erythema nodosum and hepatic granulomata. Polymerase chain reaction (PCR) was performed on biopsied liver sample to confirm the etiologic role of B. henselae and to identify the genetic variant of this organism. A PCR on the same liver biopsy for parvovirus B19 was also positive, but the clinical meaning of this was not clear.

2006 - Aberrant expression of alpha-dystroglycan in cervical and vulvar cancer [Articolo su rivista]
A., Sgambato; E., Tarquini; F., Resci; B., De Paola; B., Faraglia; A., Camerini; A., Rettino; Migaldi, Mario; A., Cittadini; G. F., Zannoni
abstract

Objectives. Cervical and vulvar cancers develop through well-defined precursor lesions but their exact pathogenesis is still unknown. The dystroglycan complex is a transmembrane glycoprotein that forms a continuous link from the extracellular matrix to the actin cytoskeleton. Deregulated expression of dystroglycan has been reported in human malignancies and related to tumor differentiation and aggressiveness. In this study, expression of dystroglycan was evaluated in the multistep cervical and vulvar tumorigenesis. Methods. Expression of the dystroglycan complex was evaluated by immunostaining in lesions representing different stages of vulvar and cervical tumorigenesis using a monoclonal antibody which recognizes carbohydratic epitopes on the alpha-dystroglycan subunit. Results. alpha-dystroglycan was constantly detected in normal cervical epithelium with a mean percentage of positive cells higher than 80%. A progressive significant reduction in the mean percentage of positive cells was observed in low (67%) and high grade SIL (14%) and in invasive carcinomas (2.6%) of the cervix. In cancers, no differences were observed in terms of percentage of positive cells when cases were stratified according with either tumor grade or stage. A progressive significant reduction in the mean percentage of positive cells was also observed from normal vulvar epithelium (90%) to VIN1 (66%), VIN2 (28%) and invasive vulvar carcinomas (22%). No significant decrease in the alpha-dystroglycan staining was observed in squamous cell hyperplasia lesions (85%) while lichen sclerosus displayed a percentage of positive cells (47%) significantly lower than normal epithelium. Conclusions. Detection of alpha-dystroglycan is frequently lost in human cervical and vulvar tumorigenesis and further studies are warranted to verify whether evaluation of this molecule might serve as marker of risk progression of preneoplastic lesions and to better understand its significance in terms of cancer development. (c) 2006 Elsevier Inc. All rights reserved.

2006 - Cell proliferation in breast cancer is a major determinant of clinical outcome in node-positive but not in node-negative patients [Articolo su rivista]
Trere, D; Ceccarelli, C; Migaldi, Mario; Santini, D; Taffurelli, M; Tosti, E; Chieco, P; Derenzini, M.
abstract

The growth rate of a tumor cell population depends on two major factors: the percentage of proliferating cells (cell growth fraction) and the rapidity of their duplication (cell proliferation rate). The authors evaluated the prognostic and predictive value of both kinetics parameters in a large series of breast cancer patients (n = 504). The cell growth fraction was determined by MIB-1 immunostaining, the cell proliferation rate by AgNOR analysis. Ki-67 LI (labeling index) and AgNOR area were significantly associated with histotype, histologic grade, tumor size, estrogen/progesterone receptor status, patient age, and lymph node involvement (P < 0.005). In the entire series of patients, both kinetics variables were significantly and independently associated with the clinical outcome, but their prognostic relevance was quite different when node-negative and node-positive patients were considered separately. Although in node-positive patients Ki-67 LI and AgNOR area were the unique independent predictors of disease-free and overall survival, they were excluded by the multivariate Cox model in node-negative patients, where only tumor size and estrogen receptor status retained a significant P-value. These results show that in breast carcinoma the cell growth fraction and the cell proliferation rate have a different prognostic impact with respect to the lymph node status and are major determinants of clinical outcome in node-positive patients only. Within this subgroup, the rapidity of cell proliferation as assessed by AgNOR analysis also served as a sensitive predictor of the response to adjuvant treatments.

2006 - Expression of the CDK inhibitor p27(kip1) and oxidative DNA damage in non-neoplastic and neoplastic vulvar epithelial lesions [Articolo su rivista]
G. F., Zannoni; B., Faraglia; E., Tarquini; A., Camerini; K., Vrijens; Migaldi, Mario; A., Cittadini; A., Sgambato
abstract

Vulvar cancer represents an important medical problem worldwide whose incidence is increasing at an alarming rate in young females. Several factors have been linked to vulvar cancer development, but its exact pathogenesis remains to be determined. Vulvar tumorigenesis proceeds through intermediate dysplastic lesions, known as vulvar intraepithelial neoplasias, frequently associated with non-neoplastic epithelial disorders of the vulva, such as lichen sclerosus and squamous cell hyperplasia. In this study, the expression of the CDK inhibitor p27(Kip1) and the extent of endogenous oxidative DNA damage were evaluated in vulvar specimens, including normal tissues, lichen sclerosus, squamous cell hyperplasia, vulvar intraepithelial neoplasias and invasive squamous cell carcinomas. We found that p27(Kip1) was constantly expressed in normal vulvar epithelium cells while a progressive significant reduction in the percentage of p27(Kip1)-positive cells was observed in vulvar intraepithelial neoplasias (77%) and in invasive carcinomas (64%). Mean percentage of positive cells in invasive carcinomas, but not in vulvar intraepithelial neoplasias, was also significantly lower than squamous cell hyperplasia lesions (78%) while lichen sclerosus displayed a percentage of positive cells (45%) significantly lower than both vulvar intraepithelial neoplasias and invasive carcinomas. 8-hydroxydeoxyguanosine (8-OHdG) is considered a sensitive biomarker for oxidative stress. We observed a progressive significant increase in the levels of 8-OHdG and in the percentage of positive cells from normal vulvar epithelium to vulvar intraepithelial neoplasias (25%) and to invasive carcinomas (64%). Squamous cell hyperplasia displayed an intermediate percentage of positive cells comparable to vulvar intraepithelial neoplasias 2 but significantly higher than vulvar intraepithelial neoplasias 1 and lower than invasive carcinomas. Lichen sclerosus staining was significantly lower than carcinomas but higher than vulvar intraepithelial neoplasias and squamous cell hyperplasia. These results demonstrate that expression of p27(Kip1) is downregulated while oxidative DNA damage increases from early non-neoplastic epithelial alterations through vulvar intraepithelial neoplasias to invasive vulvar carcinomas. Thus, both parameters might play an important role in the development of this cancer and their study might contribute to our understanding of human vulvar carcinogenesis.

2005 - Does HPV play a role in the etiopathogenesis of ameloblastoma? An immunohistochemical, in situ hybridization and polymerase chain reaction study of 18 cases using laser capture microdissection [Articolo su rivista]
Migaldi, Mario; Pecorari, M.; Rossi, G.; Maiorana, Antonino; Bettelli, S.; Tamassia, Mg; DE GAETANI, Carmela; Leocata, P.; Portolani, Marinella
abstract

Ameloblastomas are epithelial tumors of odontogenic origin, biologically characterized by local recurrence. Among different etiologic factors, HPV infection has been recently postulated to be somehow involved in ameloblastoma etiopathogenesis. To address this issue, we studied 18 ameloblastomas by means of immunohistochemistry, in situ hybridization (conventional and amplified), polymerase chain reaction and nested-polymerase chain reaction analyses using laser capture microdissection in order to detect the occurrence of HPV in this setting. No evidence of HPV infection was detected by morphological examination, immunohistochemistry, in situ hybridization and conventional polymerase chain reaction, while nested-polymerase chain reaction showed a weak positive band in two cases. However, the subsequent restriction enzyme analysis carried out from the nested-polymerase chain reaction amplification products of these two samples excluded the presence of HPV subtypes 16, 18, 31, 33, 35, 52, and 58. The search for HPV 6 and 11 in the same specimens was also negative. In conclusion, our data do not support an etiopathogenetic evidence for HPV in ameloblastoma.

2005 - PDGFR expression in differential diagnosis between KIT-negative gastrointestinal stromal tumours and other primary soft-tissue tumours of the gastrointestinal tract [Articolo su rivista]
G., Rossi; R., Valli; F., Bertolini; A., Marchioni; A., Cavazza; C., Mucciarini; Migaldi, Mario; Federico, Massimo; G. P., Trentini; A., Sgambato
abstract

Aims: To investigate the value of platelet-derived growth factor receptors (PDGFRs) by immunohistochemistry in discriminating KIT-negative gastrointestinal stromal tumours (GISTs) from other soft-tissue neoplasms of the digestive tract. Methods and results: One-hundred and sixty-seven primary gastrointestinal mesenchymal tumours (125 GISTs, 15 intra-abdominal desmoids, 12 leiomyomas, eight leiomyosarcomas, three schwannomas, two solitary fibrous tumours, and one case each of inflammatory pseudotumour and fibroid polyp) were reclassified based on morphology and on the immunohistochemical panel recommended by the National Institutes of Health consensus on GIST. All cases were then tested with antibodies specific for PDGFR alpha and beta. Of 125 GISTs, 117 were KIT-positive (93.6%) and eight KIT-negative (6.4%). All the KIT-positive GISTs were negative for both PDGFRs, while all the eight KIT-negative GISTs expressed PDGFR-alpha, with two of them also coexpressing PDGFR-beta. Among the 42 non-GIST tumours, only a small percentage (26.6%) of desmoids immunostained for PDGFR-alpha, two of them coexpressing PDGFR-beta. Conclusions: Immunostaining with PDGFR-alpha is a helpful marker in discriminating between KIT-negative GISTs and other gastrointestinal mesenchymal lesions: all KIT-negative GISTs were positive for PDFGR-alpha, while none of the other gastrointestinal mesenchymal tumours analysed, except a small subset of desmoids, was reactive with anti-PDGFRs. These preliminary data demonstrate the suitability of commercially available antibodies to detect immunohistochemically the mutually exclusive expression of KIT and PDGFR-alpha previously reported in GISTs by molecular biological techniques. Since PDGFR exists in the form of a homodimer (alpha alpha, beta beta) or heterodimer (alpha beta) and two of the KIT-negative GISTs coexpressed both PDGFR isoforms, further investigations are required to elucidate the role of PDGFR-beta in GISTs.

2005 - Post-mortem diagnosis of encephalitis in a 75-year-old man associated with human herpesvirus-6 variant A [Articolo su rivista]
Portolani, Marinella; Tamassia, Mg; Gennari, W.; Pecorari, M.; Beretti, Francesca; Alu', M.; Maiorana, Antonino; Migaldi, Mario
abstract

Abstract An HHV-6 variant A infection is described in a 75 year-old man in association with meningoencephalitis identified at autopsy. The patient presented with fever and anorexia, then he developed altered consciousness, motor weakness, progressive lethargy, and coma, and died 21 days after hospital admission. Histopathological examination showed perivascular lymphocytic infiltrates in the central nervous system (CNS). Serum and cerebral spinal fluid (CSF) samples drawn from the patient were tested for viruses by a nested polymerase chain reaction (nPCR). HHV-6 primers A and C [Aubin et al., [1991]: J Clin Microb 29: 367-372] and HS6AE and HS6AF from [Dewhurst et al. ([1993]): J Clin Microb 31: 416-418] disclosed a 750 bp genomic product of HHV-6 in both types of biological samples. Restricted site analysis showed that the HHV-6 DNA amplified belonged to the variant A of the virus. Short sequences of HHV-6 DNA could also be detected in the DNA extracted from formalin-fixed, paraffin-embedded sections of CNS tissues by use of one (GM5 and GM6) of three pairs of HHV-6 primers that were selected. Immunohistochemical examination of brain sections, employing a specific monoclonal antibody directed against the HHV-6 gp 102 protein, detected the viral antigen in neurons and glial cells. J. Med. Virol. 77:244-248, 2005. © 2005 Wiley-Liss, Inc.

2005 - Prevalence and prognostic significance of microsatellite alterations in young patients with bladder cancer [Articolo su rivista]
Migaldi, Mario; G., Sartori; G., Rossi; L., Garagnani; B., Faraglia; DE GAETANI, Carmela; A., Cittadini; Gp, Trentini; A., Sgambato
abstract

Mutations in microsatellite sequences are a hallmark of neoplastic transformation and have been reported in the majority of human cancers. Conflicting results have been reported on the role of microsatellite alterations in bladder tumorigenesis and it has been suggested that they might be mainly involved in the development of bladder cancers in young patients. In this study, DNA was extracted from laser-microdissected samples of 51 superficial papillary bladder urothelial carcinomas arising in young patients and was analyzed for the status of 19 microsatellite loci previously reported to be associated with bladder tumorigenesis. The occurrence and the pattern of microsatellite alterations, in form of loss or length variation, was evaluated and correlated with other clinicopathologic and molecular markers. The prognostic significance of these alterations was also evaluated. Loss of heterozygosity at one or more loci was detected in all 51 tumors analyzed. Length variation in at least one locus was observed in 48 (94%) of the cases. The microsatellite that was more frequently altered was D11S488 (69%), followed by D9S162 (61%), D3S3050 (55%), D3S1300 (51%) and D4S243 (51%), all the remaining being altered in less than 50% of cases. The occurrence of microsatellite alterations was not associated with tumor grade nor with tumor stage, the expression of p53, cyclin D1 or the cyclin-dependent kinase-inhibitor p27(Kip1) while it was significantly more frequent in tumors with increased expression of the proliferation marker MIB-1 (P=0.003). The occurrence of alterations at the analyzed loci was associated with a reduced risk of tumor recurrence (P=0.04 by log-rank test) and disease progression (P=0.02) in a univariate analysis. These findings demonstrate that microsatellite alterations are frequent and early events and might have a prognostic significance in bladder cancers arising at young age.

2005 - Role of chemotherapy and the receptor tyrosine kinases KIT, PDGFR alpha, PDGFR beta, and met in large-cell neuroendocrine carcinoma of the lung [Articolo su rivista]
Rossi, Giulio; A., Cavazza; A., Marchioni; L., Longo; Migaldi, Mario; G., Sartori; N., Bigiani; L., Schirosi; C., Casali; Morandi, Uliano; N., Facciolongo; Maiorana, Antonino; M., Bavieri; Fabbri, Leonardo; E., Brambilla
abstract

Purpose Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a relatively uncommon, high-grade neuroendocrine tumor sharing several features with small-cell lung carcinoma (SCLC) but currently considered as a variant of non-SCLC and accordingly treated with poor results. Little is known about the optimal therapy of LCNEC and the possible therapeutic molecular targets. Patients and Methods We reviewed 83 patients with pure pulmonary LCNEC. to investigate their clinicopathologic features, therapeutic strategy, and immunohistorchemical expression and the mutational status of the receptor tyrosine kinases (RTKs) KIT, PDGFR alpha, PDGFR beta, and Met. Results LCNEC histology predicted a dismal outcome (overall median survival, 17 months) even in stage I patients (5-year survival rate, 33%). LCNEC strongly expressed RTKs (KIT in 62.7% of patients, PDGFRa in 60.2%, PDGFR beta in 81.9%, and Met in 47%), but no mutations were detected in the exons encoding for the relevant juxtamembrane domains. Tumor stage and size (>= 3 cm) and Met expression were significantly correlated with survival. At univariate and multivariate analysis, SCLC-based chemotherapy (platinum-etoposide) was the most important variable correlating with survival, both in the adjuvant and metastatic settings (P < .0001). Conclusion Pulmonary LCNEC represents an aggressive tumor requiring multimodal treatment even for resectable stage I disease, and LCNEC seems to respond to adjuvant platinum-etoposide-based chemotherapy, Patients who received this therapy had the best survival rate. Despite our failure in finding mutational events in the tested RTKs, the strong expression of KIT, PDGFR alpha, PDGFR beta, and Met in tumor cells suggests an important role of these RTKs in LCNEC, and these RTKs seem to be attractive therapeutic targets.

2005 - Spitz nevus is relatively frequent in adults - A clinico-pathologic study of 247 cases related to patient's age [Articolo su rivista]
Am, Cesinaro; M., Foroni; P., Sighinolfi; Migaldi, Mario; Gp, Trentini
abstract

Spitz nevus is a clinico-pathologic entity that can cause diagnostic concern, particularly in adults. Many studies have been performed to establish reliable histologic criteria, in the attempt to differentiate this lesion from melanoma. A series of 247 Spitz nevi, 6 of which were formerly classified as melanomas, were reviewed for clinical and histopathological parameters. Patients older than 20 comprised 66% of cases, with a predominance of women. The lower extremity was more affected in females of any age, whereas the trunk was more frequently involved in men over 40. Histopathologic examination showed the following differences among Spitz nevi related to age: acanthosis, parakeratosis, pagetoid infiltration, and Kamino bodies were more frequent in young people, whereas multinucleated melanocytes were more frequent in adults. The latter also had lesions that were less pigmented, with less maturation and more desmoplasia. At a mean follow-up of 94 months (range 52-172), recurrence at the site of biopsy or metastases were absent. In our study, a greater proportion of Spitz nevi occurred in adults than in previous series. Moreover, the relative incidence of Spitz nevus compared with melanoma in our population was higher than in other studies. Histopathologic criteria elaborated to diagnose Spitz nevus, applied to our cases, appeared reliable, allowing a correct diagnosis, even in adults.

2004 - CDX-2 homeobox gene and MUC2 expression identifies true mucinous (so-called colloid) carcinoma of the lung and justifies its indolent outcome [Abstract in Rivista]
L., Longo; Marchioni, Alessandro; B., Murer; A., Cavazza; Losi, Lorena; Natali, Pamela; M., Bavieri; Migaldi, Mario; G., Rossi
abstract

CDX-2/MUC2 coordinated expression may be helpful in distinguishing mucinous carcinoma from mucinous bronchioloalveolar carcinoma, a lung cancer frequently associated with recurrences and multicentricity.

2004 - Primary mucinous (so-called colloid) carcinomas of the lung - A clinicopathologic and immunohistochemical study with special reference to CDX-2 homeobox gene and MUC2 expression [Articolo su rivista]
Rossi, G; Murer, B; Cavazza, A; Losi, Lorena; Natali, P; Marchioni, A; Migaldi, Mario; Capitanio, G; Brambilla, E.
abstract

Herein we describe the clinicopathologic and immunohistochemical features of 13 primary mucinous (colloid) carcinomas (MCs) of the lung, an uncommon and controversial tumor. The patients, 7 males and 6 females, ranged in age from 50 to 79 years (mean, 64.5 years). All the tumors presented as a peripheral solitary nodule with gelatinous cut-surface and well circumscribed but lacking a complete fibrous wall. The size ranged from 1 to 5.5 cm. Microscopically, they consisted of neoplastic elements floating in large mucin pools and focally lining the alveolar spaces. Eleven cases were predominantly composed of tall, columnar goblet cells (goblet cell-type MC), while 2 consisted of signet-ring tumor cells (signet-ring cell-type MC). Five tumors were incidentally discovered by chest radiographs, while the others were symptomatic. All patients underwent complete surgical resection (six lobectomies and seven wedge resections). Postoperative chemotherapy was performed in 3 cases. Overall, the median follow-up was 26 months (mean 33 months; range 9-95 months). All patients with goblet cell-type MC were alive and well, while the 2 patients with signet-ring cell-type MC died of disease. Immunohistochemically, all the 11 goblet cell-type MCs were strongly stained with CDX-2 and MUC2, 8 reacted with TTF-1, 6 with cytokeratin 20 (CK20), 9 with cytokeratin 7 (CK7), and 2 with MUC-5AC. Conversely, the two signet-ring cell-type MCs were stained with TTF-1, CK7, and MUC5AC but were negative for CDX-2, MUC2, and CK20. Surfactant apoprotein-A (SP-A) was positive in four goblet cell-type and one signet-ring cell-type MC. When compared with 10 mucinous bronchioloalveolar carcinomas (m-BAC), the latter reacted with CK7, CK20, MUC5AC, TTF-1, SP-A, CDX-2, and MUC2 in 100%, 90%, 100%, 30%, 10%, 0%, and 0% of the cases, respectively. In summary, MC of the lung represents an entity with two distinct clinicopathologic and immunophenotypic variants: 1) the goblet cell-type, presenting a more indolent clinical behavior and frequently co-expressing markers of intestinal and pulmonary differentiation; and 2) the more aggressive signet-ring cell-type, which retains only markers of pulmonary origin. On morphologic and immunohistochemical grounds, MCs are easily distinguishable from m-BAC. Since goblet cell-type MC strongly stains with CDX2, MUC2, and CK20, differential diagnosis with metastatic colorectal carcinoma is very challenging and requires appropriate clinical correlation.

2004 - Superficial papillary urothelial carcinomas in young and elderly patients: a comparative study. [Articolo su rivista]
Migaldi, Mario; Rossi, Giulio; Maiorana, Antonino; Sartori, G; Ferrari, P; DE GAETANI, C; Cittadini, A; Trentini, Gp; Sgambato, A.
abstract

OBJECTIVE: To compare the clinicopathological and immunohistochemical findings of superficial papillary transitional cell carcinomas in "young" and "elderly" patients, as the natural history and prognosis of bladder tumours in young patients remains a matter of debate. PATIENTS AND METHODS: Tumours from 50 patients with superficial urothelial tumours of the bladder diagnosed before 45 years old ("young" group, follow-up 25-119 months) were compared with 90 similar tumours developed in patients aged >55 years ("elderly", follow-up 24-102 months). All the patients had a transurethral resection with curative intent, and none had received any therapy before surgery. After surgery only patients diagnosed with pT1 tumours were treated by intravesical bacille Calmette-Guérin (BCG) instillations; all received intravesical BCG if there was a recurrence. The clinicopathological variables, recurrence and disease-free interval to recurrence were assessed. Proliferative activity (MIB-1) and expression of cell-cycle regulation proteins cyclin D1, p53 and p27(kip1) were detected by immunohistochemistry in the tumours of both groups. RESULTS There were statistically significant differences in tumour grade, stage and occurrence between the "young" and "elderly" groups. The 'young' group had a longer disease-free interval to recurrence. Among the immunohistochemical markers analysed, only MIB-1 and cyclin D1 were associated with an increased risk of recurrence in the "young" group (P < 0.04 and <0.01, respectively) in a univariate analysis. CONCLUSIONS Superficial papillary urothelial tumours of the bladder in "young" patients had a better prognosis than those in the "elderly" group, showing a lower grade and stage at diagnosis, and a lower recurrence rate. Proliferative activity and cyclin D1 expression levels were of prognostic significance for the risk of recurrence in these patients.

2004 - The prognostic role of c-kit protein expression in resected large cell neuroendocrine carcinoma of the lung. [Articolo su rivista]
C., Casali; Stefani, Alessandro; G., Rossi; Migaldi, Mario; S., Bettelli; A., Parise; Morandi, Uliano
abstract

Large cell neuroendocrine carcinoma (LCNEC) is a high-grade neuroendocrine tumor of the lung that shares some clinicopathologic and molecular features with small cell lung carcinoma (SCLC). Optimal treatment has not yet been standardized and significant prognostic factors are lacking. Because c-kit protein overexpression has been recently reported as a negative prognostic factor in SCLC we investigated its expression and prognostic value in a series of LCNEC.Resected LCNEC fulfilling the morphologic criteria of the 1999 World Health Organization classification of lung tumors and showing neuroendocrine differentiation by appropriate immunohistochemical markers were retrospectively reviewed. Immunostaining for c-kit protein expression was performed using the polyclonal antibody CD117. Clinical and pathologic characteristic were reported and analyzed and a survival study was performed.Thirty-three patients underwent radical resection. Thirty-one were male (94\%) and 32 were smokers (97\%). Ten (30.3\%), 11 (33.3\%), 5 (15.2\%), and 7 (21.2\%) were at stage IA, IB, IIB, and IIIA respectively. Overall 1-, 3-, and 5-year survival rates were respectively 79\%, 58\%, and 51\%. Survival analysis showed no differences for any of the clinicopathological features except for CD117 immunostaining: 1-year and 3-year survival rates were respectively 91\% and 82\% for CD117-negative LCNEC, and 72\% and 44\% for CD117-positive ones (p = 0.046). Positivity of CD117 was significantly related to recurrence rate: 60\% versus 23\% for CD117 positive and negative LCNEC respectively (p = 0.037).Radical resection of large cell neuroendocrine carcinoma achieves poor outcomes. The c-kit protein is frequently expressed in this neoplasia and its expression represents a negative prognostic factor. This immunohistochemical marker may represent the basic rationale to select LCNEC for novel targeted therapy.

2004 - TTF-1, cytokeratin 7, 34 beta E12, and CD56/NCAM immunostaining in the subclassification of large cell carcinomas of the lung [Articolo su rivista]
G., Rossi; A., Marchioni; M., Milani; R., Scotti; M., Foroni; A. M., Cesinaro; L., Longo; Migaldi, Mario; A., Cavazza
abstract

We selected a 4-stain immunopanel including thyroid transcription factor (TTF)-1, cytokeratin (CK) 7, 34betaE12, and CD56/neural cell adhesion molecule (NCAM) to subclassify a series of 45 pulmonary large cell carcinomas (LCCs) on bronchial biopsy. All cases consisted of a large tumor cell proliferation with abundant cytoplasm, vesicular nuclei, and prominent nucleoli. Immunohistochemically, 27 tumors (60%) were subclassified as adenocarcinoma (TTF-1+/CK7+, 24; CK7+ only, 3), 10 (22%) as squamous cell carcinoma (34betaE12+ only), and 4(9%) as LCC with neuroendocrine differentiation (CD56+, variably stained with TTF-1 and CK7, 34betaE12-). In 4 cases, the tumors coexpressed CK7 and 34betaE12 (3 cases) or were completely unstained (1 case). Surgically resected tumors matched exactly with the corresponding original biopsy specimens in 21 of 23 cases; consistent CD56 expression was a reliable marker in confirming a diagnosis of large cell neuroendocrine carcinoma even on biopsy. Our results suggest that the proposed 4-stain set of commercially available markers might help subclassify LCC even in small biopsy material, validating expression-profiling studies aimed at lung cancer classification and permitting more consistent patient enrollment for trials with targeted treatments.

2003 - Apoptosis in prostate carcinoma before and after neoadjuvant therapy with LH-RH analog [Articolo su rivista]
S., Bencini; Migaldi, Mario; G., Castagnetti; Pincelli, Carlo; P., Ferrari; DE GAETANI, Carmela; G. P., Trentini
abstract

This study investigated apoptosis in prostate cancer before and after neoadjuvant treatment with LH-RH analog, demonstrating that this therapy induced high AI and bax and survivin overexpression; thus acting as a proapoptotic agent in prostate cancer. A statistically significant correlation was found between high AI and overexpression of bax protein after therapy. It is probable that this kind of therapy is deeply implicated in promoting the apoptotic intrinsic pathway.

2003 - Dystroglycan expression is frequently reduced in human breast and colon cancers and is associated with tumor progression [Articolo su rivista]
A., Sgambato; Migaldi, Mario; M., Montanari; A., Camerini; A., Brancaccio; G., Rossi; R., Cangiano; Trentini, Giampaolo; C., Losasso; G., Capelli; A., Cittadini
abstract

Dystroglycan (DG) is an adhesion molecule responsible for crucial interactions between extracellular matrix and cytoplasmic compartment. It is formed by two subunits, alpha-DG (extracellular) and beta-DG (transmembrane), that bind to laminin in the matrix and dystrophin in the cytoskeleton, respectively. in this study we evaluated by Western blot analysis the expression of DG in a series of human cancer cell lines of various histogenetic origin and in a series of human primary colon and breast cancers. Decreased expression of DG was observed in most of the cell lines and in both types of tumors and correlated with higher tumor grade and stage. Analysis of the mRNA levels suggested that expression of DG protein is likely regulated at a posttranscriptional level. Evaluation of alpha-DG expression by immunostaining; in a series of archival cases of primary breast carcinomas confirmed that alpha-DG expression is lost in a significant fraction of tumors (66%). Loss of DG staining correlated with higher tumor stage (P = 0.022), positivity for p53 (P = 0.033), and high proliferation index (P = 0.045). A significant correlation Was also observed between loss of alpha-DG and overall survival (P = 0.013 by log-rank test) in an univariate analysis. These data indicate that DG expression is frequently lost in human malignancies and suggest that this glycoprotein might play an important role in human tumor development and progression.

2003 - Kit expression in small cell carcinomas of the lung: Effects of chemotherapy [Articolo su rivista]
G., Rossi; A., Cavazza; A., Marchioni; Migaldi, Mario; M., Bavieri; N., Facciolongo; S., Petruzzelli; L., Longo; S., Tamberi; L., Crino
abstract

A significant number of small cell lung carcinomas shows overexpression of the proto-oncogene c-kit product, a tyrosine kinase known as Kit or CD117. This molecular pathway seems somewhat implicated in promoting the neoplastic growth of small cell lung carcinoma. The current pharmacological availability of its selective inhibitor, together with the promising clinical results in the management of CD117-posidve neoplasms such as advanced gastrointestinal stromal tumors, aroused great interest among oncologists in also adopting this therapeutic strategy in other CD117-positive tumors. We evaluated a series of 27 small cell lung carcinomas, comparing the expression of CD 117 of the primary naive tumor (before first-line chemotherapy) with the expression of the same neoplasm after postchemotherapy relapse. All the patients underwent similar chemotherapeutic regimens (cisplatin/carboplatin plus etoposide). At diagnosis, 21 of 27 cases (78%) showed strong immunoreactivity for CD117. Among these 21 originally positive tumors, CD 117 remained overexpressed in 10 after relapse (48%), whereas the other 11 cases became negative. No originally CD117-negative small cell carcinomas displayed immunoreactivity after chemotherapy. CD117 expression was not statistically correlated with overall survival, occurrence of chemoresistance, or clinical response to chemotherapy. We also evaluated CD117 expression in a series of 46 surgically resected non-small cell lung carcinomas (8 squamous cell carcinomas, 10 adenocarcinomas, 5 pleomorphic carcinomas, 10 typical and 3 atypical carcinoids, and 10 large cell neuroendocrine carcinomas). Apart from small cell carcinomas, CD117 overexpression was observed in 6 of 10 large cell neuroendocrine carcinomas, whereas all the other histotypes resulted unstained. We speculate that loss of CD117 expression after chemotherapy in a high proportion of SCLC indicates that in this tumor, Kit unlikely represents the product of a constitutive mutation, as instead shown in gastrointestinal stromal tumors. Keeping this finding in mind, oncologists could re-test CD117 expression in relapsing small cell lung carcinomas in order to establish the best candidates for enrollment in ongoing clinical trials with Kit inhibitors. Practically speaking, CD 117 may be helpful in discriminating between pulmonary high-grade neuroendocrine tumors and other histotypes, but pathologists should be aware that treated small cell lung carcinomas may remain unstained in a not insignificant number of cases.

2003 - Pulmonary carcinomas with pleomorphic sarcomatoid or sarcomatous elements. A clinicopathologic and immunohistochemical study of 75 cases [Articolo su rivista]
Rossi, G; Cavazza, A; Sturm, N; Migaldi, Mario; Faccalongo, N; Longo, G; Maiorana, Antonino; Brambilla, E.
abstract

We collected 75 primary pulmonary carcinomas with pleomorphic, sarcomatoid, or sarcomatous elements to better define their clinical, histologic, and immunohistochemical profile. The patient's age ranged from 42 to 81 years (mean 65 years), and the male-to-female ratio was 9.7:1. Sixty-nine patients (92%) were smokers. Cough and hemoptysis were the most frequent presenting symptoms. Fifty-nine patients (65%) died of disease: only stage significantly predicts overall survival (p = 0.0273). Microscopically, based on the WHO criteria, 58 cases were classified as pleomorphic carcinoma (51 with an epithelial component, 7 composed exclusively of spindle and giant cells), 10 as spindle cell carcinoma, 3 as giant cell carcinoma, 3 as carcinosarcoma, and 1 as pulmonary blastoma. Immunohistochemically, in the tumors composed exclusively of spindle and/or giant cells, thyroid transcription factor-1 (TTF-1) and cytokeratin 7 were positive in 55% and 70% of the cases, respectively, whereas surfactant protein-A was always negative. In pleomorphic carcinomas with an epithelial component, cytokeratin 7, TTF-1, and surfactant protein-A were positive in the sarcomatoid component in 62.7%, 43.1%, and 5.9% of the cases, respectively, whereas they were always negative in the sarcomatous part of carcinosarcomas and blastoma. In the epithelial component of pleomorphic carcinomas, cytokeratin 7, TTF-1, and surfactant protein-A were positive in 76.4%, 58.8%, and 39.2% of the cases, respectively, whereas the same antibodies did not react with the epithelial component of carcinosarcomas; in the case of blastoma, the epithelial part of the tumor was positive for cytokeratin 7 and TTF-1, whereas it was negative for surfactant protein-A. Cytokeratin 20 was always negative. In our opinion, this study: 1) supports the metaplastic histogenetic theory for this group of tumors; 2) shows that cytokeratin 7 and TTF-1, but not surfactant protein-A, are useful immunohistochemical markers in this setting; 3) confirms that stage is at the moment the only significant prognostic parameter, as in conventional non-small cell lung carcinomas; and 4) shows that this group of tumors has a worse prognosis than conventional non-small cell lung carcinoma at surgically curable stages I, justifying their segregation as an independent histologic type in the WHO classification.

2002 - Cyclin D1 expression in papillary superficial bladder cancer: Its association with other cell cycle-associated proteins, cell proliferation and clinical outcome [Articolo su rivista]
A., Sgambato; Migaldi, Mario; B., Faraglia; G., De Aloysio; P., Ferrari; R., Ardito; DE GAETANI, Carmela; G., Capelli; A., Cittadini; Trentini, Giampaolo
abstract

Cyclin D1 contributes to regulate G1 progression by forming a complex with different cyclin-dependent kinases. It has oncogenic properties and is frequently overexpressed in several human tumor types. In our study, expression of cyclin D1 and Ki67, a proliferation marker, was evaluated by Immunohistochemistry in human papillary superficial (pTa-pT1) bladder cancers and was correlated with p27(Kip1), p21(waf1) and c-erbB-2 expression, with p53 gene status and protein expression, ploidy and cancer progression. Cyclin D1 expression was neither associated with tumor stage nor with tumor grade but high cyclin D1 expression (greater than or equal to25% positive nuclei) was significantly associated with p53 gene mutation (p = 0.012), low P21(Waf1) (p = 0.015) and high p27(KiP1) (p = 0.016) protein expression. Ki67 expression was not associated with tumor stage but a high proliferation index (greater than or equal to10% positive nuclei) was significantly associated with high tumor grade (p = 0.001) and with DNA aneuploidy (p = 0.005). There was no significant difference in proliferative activity between high and low cyclin D1 expressor tumors. Patients whose tumors showed high expression of cyclin D1 displayed a significantly longer disease-free survival (p &lt; 0.001 by log-rank test). Increased Ki67 expression was significantly associated with shorter disease-free survival (p = 0.003). Both cyclin D1 (p = 0.027; RR = 1.898) and Ki67 (p = 0.047; RR = 1.932) protein expressions were Independent predictors of reduced disease-free survival on a multivariate analysis that also included p27 Kip 1 expression and tumor stage. The simultaneous presence of low cyclin D1, low p27(Kip1) and high Ki67 expression defined a high-risk group of patients who displayed a significantly increased risk of recurrence (p &lt; 0.0001). These results suggest that evaluation of cell cycle-associated markers can help to identify high-risk patients and may affect the management of patients with papillary superficial bladder cancer.

2002 - Expression of p27(kip1) in basal cell carcinomas and trichoepitheliomas [Articolo su rivista]
Am, Cesinaro; Migaldi, Mario; S., Corrado; Maiorana, Antonino
abstract

Immunohistochemical analysis was used to evaluate p27(kip1) expression in normal hair follicles and in a series of 39 basal cell carcinomas (BCC) (13 of superficial type, 7 infiltrating, 7 morphea-like, 12 nodular) and 20 trichoepitheliomas (TE) (9 of classic type, 9 immature, 2 desmoplastic). The labeling index (LI) was derived semi automatically by means of a computer-assisted cellular image analyzer, and statistical analysis was carried out using the Student t test. A positive reaction for p27(kip1) was detected in the hair germ papillae, in supramatrical cells, and in the inner pilar sheath, whereas matrical cells and the outer pilar sheath were negative. All BCC and TE cases showed a positive immunoreaction for p27(kip1), but the staining pattern was different in the two groups of lesions, being patchy with focal peripheral accentuation in TE and more diffusely dispersed in BCC. The quantitative study showed lower p27(kip1) expression in BCC (LI = 27.51 +/- 12.55) than in TE (LI 45.27 +/- 20.27) (P &lt; 0.0001). Statistically significant differences were also observed between TE subgroups and nodular or infiltrating BCC subtypes. The Occurrence of a wide overlap of LI values hampers the practical application of a p27(kip1) LI it, the differential diagnosis between BCC and TE in difficult cases, however.

2001 - P27Kip1 expression and survival in NO gastric carcinoma. [Articolo su rivista]
Migaldi, Mario; Zunarelli, E.; Sgambato, A.; Leocata, P.; Ventura, L.; DE GAETANI, Carmela
abstract

p27Kip1, a cyclin-dependent kinase inhibitor, is considered to be a tumor suppressor gene. Absent or reduced expression of the p27Kip1 protein has been reported being a negative prognostic marker in primary lung, breast, colon, bladder, and prostate carcinomas. p27Kip1 protein expression was evaluated in a series of 96 gastric carcinomas with no lymph node involvement (NO) to verify any impact on the clinical outcome. The analysis also considered the classic clinico-pathological parameters, such as age, sex, and depth of tumor invasion (pT). The most widely used classification systems for gastric carcinoma were adopted. The expression of p27pKip1 was related neither to the pT category nor to tumor histology. Kaplan-Meier analysis documented a significant impact of an advanced pT category (p < 0.0001) and p27Kip1-reduced expression (p < 0.0002) on survival. Multivariate analysis confirmed that the reduced p27Kip1 protein expression was a strong independent predictor of poor outcome, ranking second to the pT category only (p < 0.006 and p < 0.004 respectively). As reported for other neoplasms, the expression of p27Kip1 appears to be associated with the clinical outcome of gastric carcinoma.

2000 - Apolipoprotein E polymorphism and breast carcinoma: correlation with cell proliferation indices and clinical outcome [Articolo su rivista]
E., Zunarelli; Jar, Nicoll; Migaldi, Mario; Gp, Trentini
abstract

There is preliminary evidence that polymorphism of apolipoprotein E (apoE, protein; APOE, gene), one of the key regulatory proteins in cholesterol metabolism, influences the pathobiology of carcinoma of the colon, prostate and breast and also primary tumours of the brain. This study was designed to determine whether APOE polymorphism is related to variation in the rate of tumour cell proliferation and clinical outcome in carcinoma of the breast. One hundred and eleven infiltrating ductal carcinomas, for which follow up data were available, were included in the study. Estrogen and progesterone receptor status (ER, PR) cell proliferation index (MIB-1) and APOE genotypes were determined from paraffin-embedded tissue by standard methods. Positive correlations were found between grade and tumour size, grade and presence of metastasis, grade and MIB-1 expression, as well as between ER and PR. Survival correlated inversely with tumour size and the presence of positive lymph nodes. Both steroid receptors correlated inversely with MIB-1 expression. PR positive status also correlated inversely with high histological grade and presence of lymph node metastases. APOE allele frequencies resembled those of the general population. No significant associations were found between possession of either APOE epsilon2 or epsilon4 alleles and the parameters investigated. Although there is evidence to suggest that APOE epsilon4 may predispose to the development of carcinoma of the breast our data do not support the hypothesis that APOE genotype influences the rate of tumour cell proliferation or the clinical course.

2000 - Expression of p53 and bcl-2 in clinically localized prostate cancer before and after neo-adjuvant hormonal therapy [Articolo su rivista]
Cesinaro, Am; Migaldi, Mario; Ferrari, G; Castagnetti, G; Dotti, A; DE GAETANI, C; Ferrari, P; Trentini, G. P.
abstract

The prognostic significance of p53 and bcl-2 expression in prostate carcinoma is currently under investigation. The aim of the present study was to analyze their expression in diagnostic biopsies and in prostatectomies performed after neo-adjuvant hormonal therapy to investigate their role in hormone resistance. One hundred and six patients with advanced prostate carcinoma were treated for 3 months with LHRH analogues before radical surgery. The expression of p53 and bcl-2 was analyzed by immunohistochemistry in all cases of prostatectomy and in available biopsies obtained before treatment, and was correlated with clinicopathologic parameters and follow-up. A significant increase in p53 expression was found following hormonal therapy, whereas no changes were observed in the expression of bcl-2. The increase in p53 did not correlate with the presence of therapy-induced morphological changes in prostate cancers, but it did correlate significantly with histologic grade and pathologic stage, biochemical progression of the disease, and short overall survival. At multivariate analysis, only grade and stage proved to be independent predictors of shorter survival. There were no correlations between bcl-2 and clinicopathologic variables whether in biopsies or in prostatectomies. The unfavorable clinical course associated with p53-positive carcinomas suggests that neo-adjuvant hormonal therapy may cause the selection of minor p53 mutated clones, rather than the induction of wild-type p53. In any case, the enhanced expression of p53 could label hormone-resistant cancers for further adjuvant therapy.

2000 - Loss of p21(Waf1) expression is a strong predictor of reduced survival in primary superficial bladder cancers [Articolo su rivista]
Migaldi, Mario; A., Sgambato; L., Garagnani; R., Ardito; P., Ferrari; DE GAETANI, Carmela; A., Cittadini; Gp, Trentini
abstract

P21(Waf1) is a downstream effector of p53 and belongs to the Cip1/Kip1 family of cyclin-dependent kinase inhibitors. Thus, it is a potential tumor suppressor gene and likely plays an important role in tumor development. Moreover, reduced expression of p21(Waf1) has been reported to have prognostic value in several human malignancies. In this study, we evaluated the prognostic value of p21(Waf1) in bladder cancer compared with other clinicopathological features and with p27(Kip1) and p53 expression, A total of 96 superficial (pTa-1) human bladder carcinomas were immunohistochemically stained for p21(Waf1) protein expression. Positive p21(Waf1) staining (greater than or equal to 5% positive nuclei) was observed in 68 of the 96 (71%) tumors. p21(Waf1) expression was neither associated with tumor stage (P = 0.9) nor with tumor grade (P = 0.18) but was significantly associated with both p53 protein expression (greater than or equal to 20% positive nuclei; P = 0.007) and with p53 gene mutations (P = 0.017). A significant correlation was also observed between positivity for p21(Waf1) and high (&gt;50% positive cells) p27(Kip1) expression (P = 0.04), With regard to prognosis, patients whose tumors showed absence of p21(Waf1) staining displayed a significantly shorter overall survival (P = 0.01 by log-rank test). However, p21(Waf1) expression did not correlate with disease-free survival (P = 0.15 by log-rank test). On a multivariate analysis that also included p53 and p27(Kip1) expression, negative p21(Waf1) staining was an independent predictor of reduced overall survival (P = 0.004; relative risk, 5.32), stronger than age and tumor stage. These data indicate that expression of p21(Waf1) protein strongly correlates with survival and might represent a useful prognostic marker in primary superficial bladder carcinomas.

2000 - Loss of p27(Kip1) expression is a strong independent prognostic factor of reduced survival in NO gastric carcinomas [Articolo su rivista]
A., Sgambato; Migaldi, Mario; P., Leocata; L., Ventura; M., Criscuolo; C., Di Giacomo; G., Capelli; A., Cittadini; DE GAETANI, Carmela
abstract

BACKGROUND. p27(Kip1) is a cyclin-dependent kinase inhibitor and is a potential tumor suppressor gene. Reduced expression of p27(Kip1) is a powerful negative prognostic marker in primary lung, breast, colon, bladder, and prostate carcinomas. In the current study, the prognostic value of p27(Kip1) in gastric cancer was evaluated and compared with other histopathologic parameters and p53 expression. METHODS. p27(Kip1) and p53 protein expression were determined by immunohistochemistry in 96 gastric carcinomas. The tumors were from a low incidence population and were selected for the absence of lymph node involvement. RESULTS. Reduced expression of p27(Kip1) (less than or equal to 50% positive cells) and nuclear p53 accumulation (> 30% positive cells) were observed in 67 (69.8%) and 9 (9%) tumors, respectively, and were not related to either the pT category or tumor histology. Kaplan-Meier analyses revealed a significant impact on survival by p27(Kip1) (P = 0.0001 by log rank test), p53 (P < 0.0001) expression, and the pT category (P < 0.0001). On multivariate analysis, reduced p27(Kip1) protein expression was the strongest independent predictor of reduced survival (P = 0.005; relative risk = 3.348) out weighing the pT category (P = 0.010; relative risk = 2.257) and p53 overexpression (P = 0.016; relative risk = 2.618). CONCLUSIONS. These data indicated that immunohistochemical detection of p27(Kip1) could help to identify gastric carcinoma patients who are at high risk of death, even in the absence of lymph node involvement, and who might benefit from an adjuvant treatment following surgery. Cancer 2000;89:2247-57.

2000 - Polimorfismo dell'apolipoproteina E nel carcinoma mammario: correlazione con i parametri clinico-patologici e prognosi. [Monografia/Trattato scientifico]
E., Zunarelli; Jar, Nicoll; Migaldi, Mario; Trentini, Giampaolo
abstract

There is preliminary evidence that polymorphism of apolipoprotein E (apoE, protein; APOE, gene), one of the key regulatory proteins in cholesterol metabolism, influences the pathobiology of carcinoma of the colon, prostate and breast and also primary tumours of the brain. This study was designed to determine whether APOE polymorphism is related to variation in the rate of tumour cell proliferation and clinical outcome in carcinoma of the breast. One hundred and eleven infiltrating ductal carcinomas, for which follow up data were available, were included in the study. Estrogen and progesterone receptor status (ER, PR) cell proliferation index (MIB- 1) and APOE genotypes were determined from paraffin-embedded tissue by standard methods. Positive correlations were found between grade and tumour size, grade and presence of metastasis, grade and MIB-1 expression, as well as between ER and PR. Survival correlated inversely with tumour size and the presence of positive lymph nodes. Both steroid receptors correlated inversely with MIB- 1 expression. PR positive status also correlated inversely with high histological grade and presence of lymph node metastases. APOE allele frequencies resembled those of the general population. No significant associations were found between possession of either APOE epsilon2 or epsilon4 alleles and the parameters investigated. Although there is evidence to suggest that APOE epsilon4 may predispose to the development of carcinoma of the breast our data do not support the hypothesis that APOE genotype influences the rate of tumour cell proliferation or the clinical course.

2000 - p120 expression provides a reliable indication of the rapidity of cell duplication in cancer cells independently of tumour origin [Articolo su rivista]
D., Trere; Migaldi, Mario; L., Montanaro; A., Pession; M., Derenzini
abstract

p120 is a nucleolar protein that has been immunocytochemically detected in rapidly proliferating cells of a variety of human malignancies. In the present study, the relationship between p120 expression and the rapidity of cell proliferation was evaluated in 48 human tumours of different origins. The cell proliferation rate of cancer cells was determined by quantitative analysis of AgNOR proteins. p120 immunostaining and AgNOR protein quantity,were measured by image cytometry and a highly significant correlation was found between the two variables, as evaluated by linear regression analysis (r = 0.98, p < 0.0001), The relationship between p120 expression and the rapidity of cell duplication was also studied in vitro, in six human cancer cell lines derived from different tumour types, characterized by various doubling times (ranging from 20 to 77 h), p120 expression was determined on western blots using specific anti-p120 monoclonal antibodies. Densitometric analysis revealed a highly significant inverse correlation between the integrated optical density values of the chemoluminescence bands at 120 kD and the cell line doubling times (r = - 0.93; p = 0.007), The same result was obtained bl situ by correlating p120 immunostaining of the cytological preparations obtained from the six cancer cell lines with the corresponding doubling time (r = -0.98, p < 0.0001), These results indicate that in cancer cells, the quantitative expression of p120 is directly related to the rapidity of cell duplication, independently of the tumour origin. Copyright

1999 - Correlation between high intake of glycyrrhizin and myolysis of the papillary muscles: An experimental in vivo study [Articolo su rivista]
Rossi, Tiziana; Fano, Rita Adriana; M., Castelli; M., Malagoli; Ai, Ruberto; Baggio, Giosuè Gabriele; R., Zennaro; Migaldi, Mario; Barbolini, Giuseppe
abstract

The ingestion of large quantities of glycyrrhizin, whether as a drug or st sweetener, is known, in susceptible subjects, to induce a syndrome similar to hypermineralcorticoidism, with bouts of hypertension, hypokaliaemia and rabdomyolysis, sometimes associated with severe renal failure and hypokaliaemia-induced arrythmias. Glycyrrhizin is also known to isomerize into the glycyrrhetic (or glycyrrhetinic) acids 18 alpha- and 18 beta-. In previous works, we reported that these metabolites cause bouts of hypertension and reduction in diuresis at low doses in the rat. In particular, the alpha isomer causes significant elimination of the calcium ion in the urine. The present findings confirm that 18 alpha-glycyrrhetic acid is more toxic than either glycyrrhizin or the beta isomer. Histopathological study of tissue samples taken from rats treated with the alpha isomer also reveal selective damage to the myocardium with oedema, myolysis, apoptosis and blistering of the sarcoplasm. These effects begin to appear in the course of subchronic treatment, they manifest themselves in acute treatment and correlate closely with the electrocardiographic changes recorded in rats acutely treated with 18 alpha-glycyrrhetic acid.

1999 - Detection of human papillomavirus DNA in urinary bladder carcinoma by in situ hybridization [Articolo su rivista]
DE GAETANI, Carmela; G., Ferrari; E., Righi; S., Bettelli; Migaldi, Mario; P., Ferrari; Trentini, Giampaolo
abstract

To investigate the sensitivity of an in situ hybridisation system to detect human papillomavirus (HPV) infection in transitional cell bladder cancer and to evaluate the advantages of analysing multiple biopsies; to examine the correlation between HPV tumour infection detected by in situ hybridisation and the presence of serum anti-HPV antibodies detected by enzyme linked immunosorbent assay (ELISA); and to relate the presence of viral infection to grade, stage, and follow up in cases of bladder cancer. METHODS: The in situ hybridisation technique was used with broad spectrum and type specific (6/11, 16/18, 31/33/35) probes against HPV DNA in formalin fixed, paraffin embedded tissues from 43 cases of bladder cancer. The results were analysed for the presence and type of papillomavirus and correlated with clinicopathological variables. RESULTS: The presence of HPV DNA was identified by the in situ hybridisation technique in 17 of 43 cases of bladder cancer; 12 of these were serum antibody positive and 10 had had multiple biopsies. Fifteen of the cases that were negative for HPV DNA by in situ hybridisation had positive serum serology when tested by ELISA. In 14 cases, the HPV was either types 16/18 or types 31/33/35, both of which carry high oncogenic risk. The stage (p < 0.05) and grade (NS) of the tumour and the outcome on follow up (p < 0.05) were correlated with the presence of HPV infection. CONCLUSIONS: ELISA is not useful in identifying patients with HPV positive bladder cancer, but the use of several probes and multiple biopsies increases the detection rate of HPV in neoplastic tissues. The association between tumour virus infection and high grade/high stage tumours and worse outcome suggests that HPV infection of neoplastic tissue has a negative effect on the behaviour and evolution of transitional cell bladder carcinoma.

1999 - Loss of p27(KIP1) expression correlates with tumor grade and with reduced disease-free survival in primary superficial bladder cancers [Articolo su rivista]
A., Sgambato; Migaldi, Mario; B., Faraglia; L., Garagnani; Trentini, Giampaolo; DE GAETANI, Carmela; P., Ferrari; G., Capelli; A., Cittadini; G., Romano
abstract

p27(Kip1) is a member of the Cip1/Kip1 family of cyclin-dependent kinase inhibitors and is a potential tumor suppressor gene. We previously reported a deregulated expression of p27(Kip1) in a series of human cancer cell lints and in primary breast and colon cancers. Moreover, p27(Kip1) has been reported as an important prognostic factor in primary lung, breast, colon, and prostate cancers. In this study, we evaluated the prognostic value of p27(Kip1) in a series of 96 superficial (pTa-1) human bladder carcinomas. High ( >50 % positive cells), moderate (25-50 %), and low (<25 %) p27(Kip1) staining was observed in 39 (41%), 19 (20%), and 38 (39%) of the 96 primary superficial bladder cancers, respectively. No significant association was found between the expression level of p27(Kip1) and tumor stage. Decreased p27(Kip1) staining correlated with higher tumor grade (P = 0.001). Interestingly, a significant association was observed between increased expression of p27(Kip1) and positivity for p53 (>20% positive cells; P < 0.001), A significant correlation was also observed between low expression of p217(Kip1) and decreased disease-free survival (P = 0.0003 by log-rank test) and overall survival (P = 0.01 by log-rank test), Furthermore, on multivariate analysis, low p27(Kip1) protein expression was an independent predictor of reduced disease-free survival (P = 0.018; relative risk = 1.95) second only to tumor stage, These data indicate that p27(Kip1) protein is frequently expressed at low level in poorly differentiated tumors and suggest that this protein might represent a useful prognostic marker for disease recurrence and overall survival in superficial bladder carcinomas.

1999 - Nucleolar protein p120 expression in oral carcinoma [Articolo su rivista]
L., Ventura; Migaldi, Mario; M., Criscuolo; M., Castelli; Barbolini, Giuseppe; A., Ranieri; G., Bifaretti; P., Uboldi; P., Leocata
abstract

Nucleolar protein p120 is a proliferation-associated antigen expressed by cells in early G1 phase, identified by the monoclonal antibody FB-2. Its expression has been evaluated in breast and prostate cancer; and proved to be significantly correlated with other prognostic parameters. In oral pathology, p120 protein content is able to distinguish between nonneoplastic and malignant lesions. In the present study the immunohistochemical expression of p120 protein was evaluated in fifty cases of oral squamous carcinoma and compared with histological grading, pTNM staging, DNA ploidy status and follow-lip of the patients, in order to establish its prognostic value. p120 mean area was significantly correlated to all these parameters, apart from lymph node involvement, indicating the strong predictive potential of this marker. In conclusion, quantitative immunohistochemical analysis of p120 protei;vl represents an easy and reliable method for the assessment of clinical outcome and the definition of risk groups in oral carcinoma.

1999 - The asialoglycoprotein receptor in human hepatocellular carcinomas: its expression on proliferating cells [Articolo su rivista]
D., Trere; L., Fiume; L. B., De Giorgi; G., Di Stefano; Migaldi, Mario; M., Derenzini
abstract

The expression of the asialoglycoprotein receptor (ASGP-R) on human hepatocellular carcinoma (HCG) cells might be exploited to reduce the extrahepatic toxicity of DNA synthesis inhibitors by their conjugation with galactosyl-terminating peptides. In the present study we first assessed the frequency of ASGP-R expression in 60 HCCs. Secondly, we investigated whether the receptor was maintained on the plasma membranes of DNA synthesizing cancer cells. Needle biopsies of HCC were evaluated. Diagnosis and grading of HCC were performed on routine haematoxylin and eosin-stained-sections according to Edmondson and Steiner (1953). Thirty-five tumours were grade I and II and were classified as well differentiated, while 25 tumors were grade III and IV and were classified as poorly differentiated. Sections from formalin-fixed, paraffin-embedded samples were incubated, after antigen retrieval,with an anti-ASGP-R monoclonal antibody revealed by secondary biotinylated antibody and streptavidin-biotin-peroxidase-diaminobenzidine reaction. A clear immunolabelling of plasma membranes of HCG cells was observed in 28 out of 35 (80%) well differentiated (grade I and II) and in five out of 25 (20%) poorly differentiated (grade III and IV) HCGs. The presence of the ASGP-R on the surface of DNA synthesizing cancer cells was also investigated after in vitro bromodeoxyuridine (BrdU) labelling of HCC samples by immunohistochemical visualization of both the ASGP-R and incorporated BrdU on the same section. The results obtained clearly demonstrated that DNA synthesizing cancer cells expressed the ASGP-R on their surface. The presence of ASGP-R on cell plasma membrane in the majority of differentiated HCCs and its maintenance on proliferating cells encourages studies in order to restrict the action of the inhibitors of DNA synthesis of HCC cells by their conjugation with galactosyl-terminating carriers internalized through this receptor.

1998 - AIDS splenomegaly and related iron problems [Articolo su rivista]
Ansovini, R; Barbolini, G; Migaldi, Mario; Botticelli, L; De Rienzo, B.
abstract

Spleens collected from 85 consecutive autopsies of AIDS patients (mean age 37 years) were studied. Splenomegaly, observed in 59 cases (69.4%), does not statistically correlate with life style and blood transfusions. Eleven very large spleens (over 890 g) were associated with opportunistic infections (i.e.: mycobacteria, true fungi and rochalimaea). The histological pattern was characterized by marked lymphoid depletion of the white pulp and--in 67 cases (89.4%)--packing of the pulp cords by macrophages engulfed of brown pigment which was strongly positive to the Perls reaction for ferric iron. The contemporary presence of Perls positive and p24 immunoreactive material was diffusely observed in the cytoplasm of splenic macrophages, also positive to the alkaline tetrazolium reaction. The same was observed in the cytoplasm of monocytes/macrophages of lung and brain (in 5 out of 5 patients with splenomegaly > 600 g, randomly selected). We believe that our findings deal with the formation of a haemoglobin--p24 complex and are in keeping with recent data which suggest the formation of disulphide bonds between viral proteins and haemoglobin.

1998 - p120 and AgNOR nucleolar protein expression - A comparison with nuclear proliferation markers in oral pathology [Articolo su rivista]
Migaldi, Mario; M., Criscuolo; E., Zunarelli; L., Lo Bianco; A. M., Martinelli; Barbolini, Giuseppe
abstract

To find a better method for predicting the biological behavior of certain oral cavity lesions, the expression of nucleolar protein p120 and nucleolar organizer region counts (AgNOR) was compared with that of nuclear proliferation markers MIB-1 and PCNA in 10 cases of keratotic epithelial hyperplasia (KEH), 10 cases of epithelial dysplasia (ED), and 15 cases of squamous cell carcinoma (SCC). Significant differences in p120 and AgNOR mean area values and PCNA labeling index (LI) were recorded between KEH and ED, as well as ED and SCC (Student-Neumann-Keuls test). All markers significantly differed between SCC grades I and III. Significant differences were also noted in AgNOR mean area values between grade I and II SCC and in p120 mean area values. MIB-1 and PCNA LI differed significantly when grade II and III SCC were compared (SNK test). There were significant correlations between p 120 and AgNOR (Pearson correlation coefficients) and between both of them and the proliferative indexes. AgNOR correlated with tumor grade, stage, and lymph node status (Spearman correlation coefficients), suggesting a prognostic role for that marker.

1997 - Immunohistochemical detection of nucleolar protein p120 in paraffin-embedded tissues [Articolo su rivista]
Migaldi, Mario; Barbolini, Giuseppe; D., Trere; M., Criscuolo; A. M., Martinelli; E., Zunarelli
abstract

The monoclonal antibody FB-2 recognizes the antigen p120-kDa protein (p120), associated with the nucleolar matrix. p120 has originally been reported as expressed and detectable in malignant and non-neoplastic proliferating cells, but not in most normal resting tissues and benign tumours. In the present study, a reliable immunostaining method was used to detect p120 on formalin-fixed, paraffin wax-embedded tissue, testing it on 148 samples from different neoplastic and non-neoplastic tissues from different organs (breast, colon, lung, prostate, bladder, lymph nodes, skin, tongue and liver). The immunostaining was performed after the application of a specific antigen-unmasking protocol based on six consecutive cycles of microwave oven heating. Under these retrieval conditions, p120 antigen was clearly detectable, not only in hyperplastic and malignant cells, but also in stromal and normal non-proliferating cells of all the tissues evaluated. Our results show that the nucleolar protein p120 can be detected by routine immunohistochemistry in formalin-fixed, paraffin-embedded tissue and is expressed in all nucleated cells under any biological condition.

1997 - Relationship between quantitative expression of protein p120 and proliferative activity in cancer cells [Articolo su rivista]
Trerè, D; Montanaro, Laura; Migaldi, Mario; Pession, A; Derenzini, M.
abstract

In questo articolo, il nostro gruppo tra i primi a livello europeo indirizza la ricerca verso la p120, proteina nucleolare con notevoli potenzialità sulla cinetica di crescita tumorale.

1996 - Bulletin of breast disease: results of the 1990-1994 quinquennium [Articolo su rivista]
Criscuolo, M; Migaldi, Mario; Trentini, G. P.
abstract

All mammary lesions diagnosed at the Institute of Pathological Anatomy of the University of Modena have been systematically filed since 1990 and reported in a bulletin, which is issued twice a year and delivered to health operators. So far, 5.188 cases of breast lesions, comprising 1.999 non-neoplastic pathologies, 1.040 benign tumors, 1.943 primary malignant neoplasms and 206 recurrences, have been filed. During the quinquennium 1990-1994, a progressive numerical reduction in diagnoses of non-neoplastic lesions coupled to an increase of benign tumors has been observed, whereas the number of primary malignant tumors has not changed. In particular, a statistically significant increase in diagnoses of carcinoma-in-situ and of fibrocystic disease associated with moderate-risk lesions (atypical hyperplasias) has been detected, whereas the number of cases of single fibrocystic disease has decreased. This reduction, however, is not significant. A slight increase of breast carcinomas smaller than 1 cm and 2 cm, coupled to a decrease of those exhibiting dimensions between 2 and 5 cm, has been found. The collection and systematic analysis of cases of mammary lesions appears to represent a useful tool to study the incidence of different breast pathologies in the general populations. It can also be viewed as a simple way to test the reliability of diagnostic methods used for selection of surgical cases.

1996 - The changing histopathology of primitive lung cancer. [Articolo su rivista]
Migaldi, Mario; Zunarelli, E; Criscuolo, M; Lauriola, P; De Girolamo, G; Barbolini, G.
abstract

his study was performed on 1710 lung epithelial malignant tumours, diagnosed in the biennia 1963/1964 (42 cases), 1973/ 1974 (293), 1983/1984 (637) and 1993/1994 (738). It was aimed at evaluating whether, over a time span that long, the distribution of the histopathological patterns could have changed. The neoplasms were classified according to the WHO criteria (1981). From our data, a striking increase becomes apparent for all histotypes. Squamous cell carcinoma holds the leading position both among males and females, even though its rate of increase tends to slow down among males. Analogous trend is observed for all the other histotypes but adenocarcinoma. Among females, the rate of increase continues to accelerate for all histotypes but large cell carcinoma. The male to female ratio is ranging from 2:6 for adenocarcinomas to 12:1 for squamous cell carcinomas. In conclusion, impressive changes seem to have occurred in the frequency of the different lung oncotypes over this time span, especially for adenocarcinoma (increasing trend) and large cell carcinoma (decreasing trend). It also seems worth underlying that the marked increase among females could make lung carcinoma the number one cause of cancer-related deaths in this sex.

1995 - Argyrophilic nucleolar organizer regions and bromodeoxyuridine and h3-thymidine labelling indices in colorectal cancer [Articolo su rivista]
Losi, Lorena; C., Di Gregorio; R., Fante; Migaldi, Mario; Roncucci, Luca; Pedroni, Monica; PONZ DE LEON, Maurizio; G. P., Trentini
abstract

The count of argyrophilic nucleolar organizer regions (AgNORs) has been proposed as a useful method for evaluating cell replication in human tumours. The current study was undertaken to compare AgNOR values in colorectal cancers with two better established methods for investigating cell proliferation such as bromodeoxyuridine (BrdUrd) and (3)[H]-thymidine ((3)[H]dT) labelling indices (LIs). Because some concern still exists regarding accuracy and reproducibility of AgNOR quantifying methods, we carried out a control study by independently repeating the same measurements (number, area and area per silver-stained NOR particle) in two centres with different operators and computer-assisted image analysers on 40 colorectal carcinomas. AgNOR values recorded in the two centres were strictly correlated (r = 0.75; P &lt; 0.001 for number; r = 0.62, P &lt; 0.01 for area; r = 0.63, P &lt; 0.001 for area per silver-stained NOR particle) and the range of values were almost identical, Then, AgNOR values were compared with BrdUrd and (3)[H]dT LIs, respectively obtained by in vivo incorporation and in vitro incubation in the same series of colorectal carcinomas. No correlation was found between AgNOR values and BrdUrd or (3)[H]dT LIs. BrdUrd and (3)[H]dT LIs were instead reciprocally significantly correlated, No evident correlation was seen between LIs or AgNOR values and clinico-pathological parameters of the tumour. In conclusion, in colorectal neoplasms, AgNOR values did not appear to relate with more direct parameters of cell proliferation. It follows that AgNOR reliability as a biomarker of cell proliferation remains questionable.

1995 - HIGHER REPRODUCIBILITY OF MORPHOMETRIC ANALYSIS OVER THE COUNTING METHOD FOR INTERPHASE AGNOR QUANTIFICATION [Articolo su rivista]
Trere, D; Migaldi, Mario; Trentini, Gp
abstract

In a series of 40 breast carcinomas, the reproducibility of two different methods for interphase AgNOR quantification was evaluated. Two operators independently defined on each slide the interphase AgNOR quantity both by measuring the area of the silver-stained structures using image cytometry and counting the AgNOR number directly at the microscope. The correlation between the values obtained by the two observers was statistically significant, but the correlation coefficient between AgNOR areas (r = 0.79; P < 0.001) was greater than that between AgNOR numbers (r = 0.38; P = 0.014). On the other hand, when interphase AgNOR area and number values obtained by each observer were compared, no significant correlation was found. This study has demonstrated that the two different methods for interphase AgNOR quantification are not comparable, and that morphometric analysis is more objective and reproducible than the counting method.

1995 - Metastatic breast carcinoma presenting as urinoma. [Articolo su rivista]
Zunarelli, E; Pollastri, Ca; Calo, M; Migaldi, Mario; Galizia, G; Criscuolo, M.
abstract

A rare case of breast carcinoma metastatising to ureter and presenting as urinoma is described. Reviewing the literature, only few cases of urinoma due to metastasis from breast carcinoma have to date been reported. The diagnostic role of immunohistochemical study in defining the disease is discussed.

1995 - Quantitative and qualitative AgNORs rates of prostate cancer on needle core biopsies: a multicentric study. [Articolo su rivista]
Botticelli, Ar; Marandola, P; Jallous, H; Vicini, D; Migaldi, Mario; Speroni, A; Mirando, P.
abstract

From 1992 to 1993, the Scarpa Foundation Center of Pavia (SFCP) with 12 associated Italian Urological Units selected 40 cases of prostate diseases discovered on needle core biopsies, 5 of which were benign hyperplasias (BPH) in patients without clinical and morphological evidence of cancer and 35 prostate cancers (PRC) classified according to Gleason's histological grades (GLG) of PRC malignancy. Serum prostatic specific antigen (PSA) values were tested before clinical urological examination or biopsy or surgery. In all groups, AgNORs scores/nucleus were obtained by semi- and -automatic computerized image analysis and also by qualitative subjective counts of three observers on light microscopy. Our results pointed out a good correlation between PSA levels, GLG of PRC malignancy and AgNORs scores. The quantitative method showed an average number of AgNORS dots per nucleus between 2 and 3 in well differentiated PRC and higher than 3 in moderately differentiated and undifferentiated PRC, and exhibited more sensitivity over GLG3 than the qualitative investigation. The qualitative subjective count of AgNORs dots/nucleus seemed to be more reliable in differentiating the AgNORs scores of BPH (average of 1.81 dots/nucleus) from very well differentiated PRC with GLG1 + 2(average of 2.25 dots/nucleus) than quantitative analysis, which showed the same average value in both groups (2.11 dots/nucleus). For these reasons, also on needle biopsies of benign and malignant prostate diseases the subjective AgNORs count may aid the histological diagnostic judgement of malignancy, by avoiding misleading diagnoses of microscopic pictures of BPH cancer look likes and a predictive histologic malignant factor, in identifying PRC with low or high progression.

1994 - ONTOGENY OF THE CIRCADIAN-RHYTHM IN MEDIAL BASAL HYPOTHALAMIC BETA-ENDORPHIN CONTENT IN FEMALE RAT [Articolo su rivista]
Criscuolo, M; Degaetani, C; Ficarra, G; Nappi, Re; Migaldi, Mario; Petraglia, F; Genazzani, Ar; Trentini, Gp
abstract

The present study evaluated the possible role of estrogens in generating the circadian rhythm of medial basal hypothalamus content at the time of puberty in female rats. Accordingly, changes in medial basal hypothalamus beta-endorphin (beta-EP) content were investigated in female rats, before and at puberty. Groups of intact or ovariectomized rats were studied after estradiol-benzoate or placebo treatment. The results showed that circadian rhythm of beta-EP content of medial basal hypothalamus is absent in prepubertal rats, while it appears at puberty, associated to a significant increase of beta-EP concentration. The primary involvement of steroids in generating this circadian rhythm was supported by the finding that estradiol-benzoate treatment caused a precocious appearance of beta-EP hypothalamic diurnal changes in prepubertal rats. Moreover, estradiol-benzoate replacement restored the loss of beta-EP nocturnal increase induced by ovariectomy in pubertal animals. Therefore, these data support the significant role of estrogen in inducing the circadian rhythm of beta-EP content in medial basal hypothalamus at the time of puberty in female rats.

1994 - Prognostic significance of nucleolar organizer regions in recurrent pleomorphic adenomas of salivary glands. [Articolo su rivista]
Criscuolo, M; Migaldi, Mario; Collina, G; Cesinaro, Am; Galetti, R; Lo Bianco, F; Trentini, G. P.
abstract

Pleomorphic adenoma (PA) is the commonest tumor of salivary glands. It is considered a benign tumor which may recur and rarely metastatize. There are no reliable histological criteria for predicting recurrences of PA. In this paper, Ag-NOR expression, evaluated by means a computer assisted image analysis system, has been tested on 11 cases of PA, their recurrences, 10 non recurrent PA and 5 cases of carcinoma ex PA. Mean AgNOR count, mean AgNOR area and mean AgNOR area/particle were determined in each group. AgNOR count and area did not show any significant differences between recurrent and non recurrent PA. On the contrary, AgNOR area/particle seems to be a useful tool in predicting the clinical behaviour of this lesion.

1994 - Salivary gland tumors: revision of 391 cases according to the new WHO classification. [Articolo su rivista]
Cesinaro, Am; Criscuolo, M; Collina, G; Galetti, R; Migaldi, Mario; Lo Bianco, F.
abstract

We reviewed 391 cases of salivary gland tumors collected in a 13 year period at the Institute of Pathological Anatomy of the University of Modena, according to the new WHO classification (1991). A diagnosis of pleomorphic adenoma was changed in adenoid cystic carcinoma. Within malignant tumors, 16 cases were reclassified, and new entities, such as polymorphous low-grade adenocarcinoma, papillary cystoadenocarcinoma and salivary duct carcinoma were introduced. Mucoepidermoid carcinomas were subdivised in low-grade and high-grade tumors, but at the follow-up histological grade did not correlate with clinical behaviour of the lesions.

1993 - PROGNOSTIC-SIGNIFICANCE OF NUCLEOLAR ORGANIZER REGIONS IN OVARIAN EPITHELIAL TUMORS [Articolo su rivista]
Criscuolo, M; Martinelli, Am; Migaldi, Mario; Zunarelli, E; Bergamaschi, M; Falchi, Anna Maria; Degaetani, C.
abstract

Recent studies have shown that the silver-stained nucleolar organizer region (AgNOR) score is related to the cell growth rate in several neoplasms. In the work presented here, we tested the AgNOR technique in 79 ovarian epithelial tumors (13 benign, 10 borderline, 56 malignant) to evaluate the diagnostic potential of AgNOR count in distinguishing between ovarian borderline tumors and carcinomas and to assess its prognostic value in carcinomas. Ovarian carcinomas exhibited higher mean AgNOR values than borderline and benign tumors, but statistically significant differences were found only in the serous type. Statistical analysis showed a positive correlation of higher AgNOR score, advanced tumor stage, and adverse prognosis. On the contrary, low AgNOR counts identified stages I and II carcinomas with disease-free follow-up. These results suggest that the AgNOR count may improve the prognostic evaluation of ovarian epithelial tumors by representing a reliable indicator of survival.

1993 - PROLIFERATING CELL NUCLEAR ANTIGEN CYCLIN IN INCIDENTAL CARCINOMA OF THE PROSTATE [Articolo su rivista]
Botticelli, Ar; Criscuolo, M; Martinelli, Am; Botticelli, L; Filoni, A; Migaldi, Mario
abstract

Monoclonal antibody to proliferating cell nuclear antigen (PCNA) has been used to identify the growth fraction in ten cases of benign prostatic hyperplasia (BPH), in 20 prostatic microcarcinomas (PMC) and in 30 cases of infiltrating prostatic carcinoma (PC). Ten year follow-up was available on all cases by means of clinical, serological, radiological and echographic examinations. The percentage of PCNA-staining nuclei was independently counted by two observers. Statistical analysis showed significant differences between PCNA/cyclin score of BPH and PMC without recurrences with respect to those of PMC with progression and of PC. PCNA immunostaining may represent a reliable method for assessing cellular proliferative activity. It may be used as a more powerful diagnostic hallmark of PMC than patterns of non-malignant microglandular proliferation and is also a useful additional test for assigning histological grades to PMC and PC. Statistical analysis indicated that PCNA/cyclin index was an independent significant prognostic indicator of predicting malignant progression (P less-than-or-equal-to 0.01) and survival rates (P less-than-or-equal-to 0.05) of PC and PMC (> 5 mm diameter).

1992 - Melatonin treatment delays reproductive aging of female rat via the opiatergic system. [Articolo su rivista]
Trentini, Gp; Genazzani, Ar; Criscuolo, M; Petraglia, F; De Gaetani, C; Ficarra, G; Bidzinska, B; Migaldi, Mario; Genazzani, Alessandro
abstract

In female rat age-related reproductive decline is accompanied by progressive impairment of the neuroendocrine mechanisms that regulate LH secretion. The biosynthetic activity of the pineal gland is markedly depressed and the nocturnal secretion of melatonin decreases significantly. The aim of the present study was to evaluate whether the nocturnal administration of melatonin via the drinking water (0.4 micrograms/ml) throughout the course of aging from 14 to 24 months of age could (1) influence the age-related changes that occur in basal serum levels of LH and in the LH response to GnRH or to naloxone stimulation at 16, 18 and 20 months of age, and (2) delay the onset of the postreproductive constant estrous-anovulatory state as evaluated by the daily recording of vaginal smears and by occurrence of polyfollicular ovaries at 24 months of age. Our results demonstrate that melatonin replacement delays the increase in LH serum levels and the decrease in LH response to GnRH that occur in 18-month-old control animals. Furthermore, they show that melatonin treatment prevents the loss of LH response to naloxone manifested in control rats between 16 and 20 months of age. Melatonin also appears to prevent the progressive increase in the monthly occurrence of estrus phases as well as to decrease the number of rats with polyfollicular ovaries at 24 months of age in comparison to control animals. These results suggest that the age-related decrease in circulating melatonin during the night may contribute to the reproductive decline of aging, and that this effect may involve the central opioid system.

1991 - Pathological findings in perinatal autopsies [Capitolo/Saggio]
Criscuolo, M; Migaldi, Mario; Botticelli, Annibale Renzo; Losi, Lorena
abstract

The study of perinatal mortality rates has recently be entered by pathologists, because of their capacity to discover the true cause of death at necroscopy.

1989 - Immunohistologic analysis of mycobacterial antigens by monoclonal antibodies in tuberculosis and mycobacteriosis. [Articolo su rivista]
Barbolini, G; Bisetti, A; Colizzi, V; Damiani, G; Migaldi, Mario; Vismara, D.
abstract

Four monoclonal antibodies (MoAbs), 60.15, 61.3, 105.10, and 2.16, directed to different proteins of Mycobacterium tuberculosis were used by an indirect avidin-biotin complex peroxidase-antiperoxidase method to detect mycobacterial antigens in lung, lymph node, and joint tissue specimens of tuberculous patients. Using MoAb 60.15, which recognizes a broad range of cross-reactive mycobacterial proteins with a molecular mass of 28 kilodaltons (kD), scattered materials (mycobacterial in origin) were observed, many of which were located within phagocyte cytoplasm. With MoAb 61.3, which reacts with a 35 kD protein present in M tuberculosis, Mycobacterium africanum, and Mycobacterium bovis, many clumped particles similar in size and shape to acid-fast bacilli were observed within the phagocyte cytoplasm (lung tissue) and positive macrophages with lysosomes were distributed throughout the cytoplasm (bronchoalveolar lavage). The specificity of this MoAb (61.3) was confirmed by the negative staining of positive lymph node specimens obtained from a patient infected with Mycobacterium kansasii. MoAbs 105.10 and 2.16 bind to the cross-reactive 65 kD heat shock protein that is present in mycobacteria and stain scattered particles and dark clumps of bacilli within the phagocyte cytoplasm. On the basis of this study, immunohistochemical detection of mycobacterial antigens appears to be useful in establishing the mycobacterial etiology of caseating granulomas and in avoiding the false-negative results obtained by traditional staining methods.