M Castagna

Università di Pisa, Pisa, Tuscany, Italy

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Publications (44)152.82 Total impact

  • A Failli · A Legitimo · G Orsini · M Castagna · R Spisni · P Miccoli · R Consolini ·
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    ABSTRACT: Adjuvant therapy in colorectal cancer has evolved to become the standard of care, whereas the tumor capability of activating effective mechanisms of defence against both chemical and physical cytotoxic agents represents a serious obstacle to the successful therapy of human tumors. Therefore, the possibility to have an assay useful to measure the drug sensitivity of tumor cells has a great importance. A number of cytotoxicity assays are currently available, each of them using a specific approach to detect different aspects of cell viability, such as cell integrity, proliferation and metabolic functions. The purpose of this study is to compare, under identical experimental conditions, three common cytotoxicity assays (ATP-lite, MTT and CCK-8 assays) in the assessment of the anti-proliferative effects of 5-fluorouracil (5-FU) and oxaliplatin (OHP) on three colon cancer cell lines (WiDr, SW620 and HT-29). Regarding 5-FU, the three assays were found to be significantly correlated with a moderate or high correlation coefficient, whereas in the case of OHP we found different outcomes among the assays. Our study demonstrates that the CCK-8 is the most sensitive assay for detecting changes of cell viability, suggesting that the viability measured in cells after drug exposure depends on several parameters like the drug used, the biological characteristics of the target cell and the specific approach employed by the method to detect distinct cell growth and metabolic functions.
    Journal of biological regulators and homeostatic agents 03/2013; 27(1):275-284. · 2.04 Impact Factor
  • A Iannelli · C Martini · M Cosottini · M Castagna · F Bogazzi · L Muscatello ·

    Child s Nervous System 11/2011; 28(2):297-303. DOI:10.1007/s00381-011-1626-3 · 1.11 Impact Factor
  • A Failli · R Consolini · A Legitimo · G Orsini · A Romanini · R Spisni · M Castagna · P Miccoli ·
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    ABSTRACT: Adjuvant therapy has evolved to become the standard care of colon cancer, but the tumor capability of activating effective mechanisms of defence against both chemical and physical cytotoxic agents represents a serious obstacle to the successful therapy. Furthermore, the possibility to have an assay useful to measure the drug sensitivity of tumor cells could be of a great importance. As primary human colon cancer cultures from fresh tumor are technically difficult to obtain, experiments with human cancer cell lines remain essential to explore new adjuvant chemotherapy drugs, to investigate the individual responsiveness to the known agents, and particularly to clarify how these chemotherapeutic agents could be used in maximizing outcomes. In the present study we evaluate the cytotoxic effects of 5-fluorouracil (5-FU) and oxaliplatin (OHP) and of their pharmacological interaction in three human colon cancer cell lines (WiDr, HT-29 and SW620), by using an ATP luminescence assay (ATPlite; Perkin Elmer), displaying high sensitivity, linearity and reproducibility. Cell cycle, apoptosis and CD44 expression were investigated with flow cytometry. Our results show that the drug combinations inhibited the cell growth more than each drug alone in all colorectal cancer cell lines. Interestingly, the sequential exposure of OHP and 5-FU resulted in the most cytotoxic effect in all colon cancer cell lines, when compared to the simultaneous one. Our results focus on the powerful cytotoxic effect of 5-FU-OHP combination, when used in sequential exposure, suggesting interesting implications for a rational use of 5-FU, OHP combination in colon-rectal cancer therapy.
    Journal of biological regulators and homeostatic agents 10/2011; 25(4):575-88. · 2.04 Impact Factor

  • Neuropathology and Applied Neurobiology 02/2008; 34(5):564-8. DOI:10.1111/j.1365-2990.2007.00934.x · 3.93 Impact Factor
  • S Romoli · A Abbruzzese · M Castagna · F Becherini · G Parenti ·
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    ABSTRACT: Subependymoma was first described by Scheinker in 1945; it frequently occurs in the ventricles and rarely in the spinal canal representing 0.7% of all central nervous system tumours. Most of these intraventricular tumours are subclinical entities, remaining of small size and discovered at autopsy with 0.4%incidence. We report a case of subependymoma with a completely exophytic growth from the foramen of Luscka: only a similar one has been described in the literature but with a lesser cysternal involvement. Neuroradiological and anatomopathological features of subependymoma are discussed.
    Journal of neurosurgical sciences 07/2007; 51(2):81-4. · 1.16 Impact Factor
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    ABSTRACT: Prostaglandins regulate various functions throughout the gastrointestinal system. Their biosynthesis depends on cyclooxygenase isoforms, named COX-1 and COX-2. The initial hypothesis that COX-2 is an inducible enzyme has been challenged and its constitutive expression in the stomach has been established. In this study, an immunohistochemical analysis was performed to evaluate the distribution and cellular localization of COX-2 in normal human colon. Colonic surgical specimens were processed for COX-2, protein HuC/HuD, neurofilament, S-100 protein and CD117/c-kit immunodetection. COX-2 protein was found to be constitutively expressed in the colonic wall: detectable amounts were localized in mucosal, submucosal and muscular layers, mainly in the neuromuscular compartment. In particular, COX-2 was expressed in muscularis mucosae, submucosal ganglia, longitudinal muscle layer and myenteric ganglia, the neurons of which displayed different degrees of immunostaining. Intramuscular interstitial cells of Cajal, regarded as important sites for the regulation of enteric neuromuscular activity, were also partly COX-2 immunoreactive. This study provides a detailed mapping of COX-2 expression in human colon, and allows better understanding of the roles played by this isoenzyme in gut physiology.
    Neurogastroenterology and Motility 09/2006; 18(8):654-62. DOI:10.1111/j.1365-2982.2006.00795.x · 3.59 Impact Factor
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    ABSTRACT: This study describes a case of autochthonous neurocysticercosis in a non endemic region where a differential diagnosis with more frequent single parenchymal lesions must be carried out. The patient presenting generalized seizures and coma status under the suspicion of cerebral neoplasia was admitted to the neurosurgery division. MRI showed the presence of an ovoidal cystic lesion in right-frontotemporal region. After right fronto-temporal osteoplastic bone flap elevation and usual dural tacking and opening, trans-scissural subaracnoid access was made possible by microsurgical strumentation in right Sylvian fissure. A cystic, translucid lesion was identified in the deep Sylvian fissure, involving the distal segment of the fissure and the frontal lobe, the cystic lesion was removed surgically. Pathological examination showed a typical picture of neurocysticercosis. After both surgical and chemotherapeutic treatment with albendazole the outcome was successful. The diagnosis of neurocysticercosis should be taken into account in the presence of generalized seizures possibly due to single parenchymal lesions, even in non-endemic regions for this parasitic infection.
    Pathologica 09/2006; 98(4):229-31.
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    ABSTRACT: This study was designed to determine the debated prognostic significance of reverse transcriptase-polymerase chain reaction (RT-PCR) positivity in melanoma patients' sentinel lymph node (SLN) negative by conventional histopathology (PATH). Patients with primary stage I-II cutaneous melanoma underwent radioguided sentinel lymphadenectomy. Their SLNs were assessed for tyrosinase (Tyr) and melanoma antigens recognized by T-cells (MART-1) mRNA expression using RT-PCR, in parallel with hematoxylin and eosin staining and immunohistochemistry. Tyr and MART-1 expression in the SLNs were correlated with PATH assay results, standard prognostic factors, time to progression and overall survival. Twenty-three of the 124 patients (18.5%) had positive SLNs by both PATH and RT-PCR (PATH+/PCR+). Sixteen patients (13%) were negative by PATH and positive by RT-PCR (PATH-/PCR+). Eighty-five patients (68.5%) had SLNs that were negative by both PATH and RT-PCR (PATH-/PCR-). At a median follow-up of 30 months, recurrence rates among the three cohorts were statistically different (PATH+/PCR+, 60%; PATH-/PCR+, 31%; PATH-/PCR-, 9.4%). Seven of 23 (30%) and two of 16 (12.5%) patients died in the PATH+/PCR+ and PATH-/PCR+ SLN groups, respectively, whereas no patient died in the PATH-/PCR- SLN group. RT-PCR is more sensitive than PATH to detect SLN metastases and it is a reliable predictor of disease relapse in stage I-II melanoma patients.
    Annals of Oncology 12/2005; 16(11):1832-40. DOI:10.1093/annonc/mdi372 · 7.04 Impact Factor
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    ABSTRACT: In this study we report the presence of peripheral benzodiazepine receptors (PBRs) in human parathyroid glands and describe the effect of their benzodiazepine type ligands on parathyroid cell function. PBR binding features in normal parathyroid tissue were characterized and compared to parathyroid adenoma, using a specific and selective ligand for PBR, [3H] 1-(2-chlorophenyl)-N-methyl-N-(1-methyl-propyl)-3-isoquinoline-carboxamide ([3H]PK11195). Affinity and density of [3H]PK11195 binding sites in homogenate membrane preparations from adenomatous and normal tissues were determined. Parathyroid adenoma showed a statistically significant 2.2 fold increase of [3H]PK11195 binding sites, while the affinity remained unchanged. Our results represent the first evidence of PBRs in parathyroid glands and suggest for them a role in influencing PTH release. A clear trend of PBR up-regulation in parathyroid adenoma was also found.
    Journal of endocrinological investigation 11/2004; 27(9):826-31. DOI:10.1007/BF03346276 · 1.45 Impact Factor

  • Annual Meeting, Society of Nuclear Medicine, USA; 05/2002
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    ABSTRACT: In the 1990s, the introduction of the Guglielmi detachable coil (GDC) system in clinical practice was followed by extensive clinical use of this endovascular device in the treatment of brain aneurysms. This technology is based on electrothrombosis and electrolytic detachment of platinum coils. Despite the extensive use of this treatment technique, the role of electrothrombosis has not been fully investigated and clarified. An in vitro electron microscopic study of human blood was performed to elucidate the role that electrothrombosis might play in triggering the biologic response of thrombosis of the aneurysmal sac. Human blood from five patients was used to fill plastic containers in which GDCs had been deposited. These five patients had subarachnoid hemorrhage and were similar in age and clinical presentation. Electron microscopic studies were performed on GDCs that had been electrically charged and on GDCs that had not. All electron microscopic studies revealed that the electrically charged GDCs were covered by blood elements and fibrin adherent to the surface of the coil. Noncharged GDCs did not have deposits or adhesions of these blood constituents. These findings demonstrated that passage of electric current through the GDC induces attraction of blood constituents. This attraction may trigger a thrombotic reaction on the surface of the coil. The greater the time of current application, the more pronounced the cellular reaction and the deposition of fibrin and blood cells on the GDC.
    American Journal of Neuroradiology 11/2001; 22(9):1757-60. · 3.59 Impact Factor
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    ABSTRACT: Thyroid hormone is essential for maintaining normal neurological functions both during development and in adult life. Type III-iodothyronine deiodinase (D3) degrades thyroid hormones by converting thyroxine and 3,5,3'-triiodothyroinine (T3) to inactive metabolites. A regional expression of D3 activity has been observed in the human central nervous system (CNS), and a critical role for D3 has been suggested in the regulation of local T3 content in concert with other enzymes. This study was undertaken to further characterize D3 activity in human CNS and to understand its role in the local regulation of T3 content. Autoptic specimens from various areas of human CNS were obtained 6--27 h postmortem from 14 donors who died from cardiovascular accident, neoplastic disease or infectious disease. D3 was determined by measuring the conversion of T3 to 3,3'-diiodothyronine. The T3 content was measured by radioimmunoassay in ethanol extracts, using a specific antiserum. High levels of D3 activity were observed in hippocampus and temporal cortex, lower levels being found in the thalamus, hypothalamus, midbrain cerebellum, parietal and frontal cortex, and brain stem. An inverse relationship between D3 activity and T3 content in these areas was demonstrated. We have concluded that D3 contributes to the local regulation of T3 content in the human CNS.
    European Journal of Endocrinology 07/2001; 144(6):577-83. DOI:10.1530/eje.0.1440577 · 4.07 Impact Factor
  • S Pistolesi · M Castagna · R Vannozzi · R Cantini · R Pingitore ·
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    ABSTRACT: A case of neuroglial tumor in a 18-year-old man is presented. The neoplasm was composed by two cell types. One type showed features typical of neuronal cells, while the other resembled glial cells. The diagnosis was confirmed by immunohistochemistry results.
    Pathologica 03/2001; 93(1):57-60.
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    ABSTRACT: Carcinoma of the rectal colon begins as a small neoplastic polyp which gradually increases in size and, after passing through various degrees of dysplasia, develops into an overtly malignant carcinoma. Clinical experience suggests that patients may be divided into subgroups based on the aggressivity of the tumour. The genetic mutations associated with colorectal cancer have been studied and it is known that the genes primarily responsible for biological changes in the tumour cell, in the early stages, are APC, hMSH2, k-ras2 and, in particular, p53. Indeed, the mutation at the level of gene p53 has been recognized as the most common mutation in tumour cells. The aim of this study was investigate the role of p53 and CD34 in colorectal cancer. We studied p53 positivity using immunohistological methods and compared our results with the site, stage (using the TNM system) and histological grade of the tumour. We evaluated CD34 positivity using the same methods in order to detect and quantity the presence of angiogenesis in colorectal cancer. P53 was found to be markedly raised in the T3 stage of colorectal cancer, while its expression was decreased in stage T2 and stage T1 carcinomas and it was not detectable in adenomas. These results suggest a close correlation between the tumour stage and the expression of p53. An analogous correlation was found between CD34 expression and angiogenesis. The overexpression of p53 in epithelial cells and raised angiogenesis (as reflected in CD34 levels) in stromal cells could represent useful prognostic factors in the management of colorectal cancer.
    Annali italiani di chirurgia 01/2001; 72(3):323-7. · 0.60 Impact Factor
  • B Loggini · I Rinaldi · R Pingitore · R Cristofani · M Castagna · P Barachini ·
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    ABSTRACT: Thickness and level of invasion are the main morphological elements for an approximate but not sufficiently sensitive prognostic evaluation of cutaneous melanomas. By using immunohistochemical methods it is possible to detect biological markers related to prognosis. We have studied p53, PCNA, Bcl-2 and P-gp expression in 49 primary cutaneous melanomas. We used the immunophosphatase APAAP immunohistochemical method. The percentage of labeled cells (according to four classes of positivity: <5%; 5-25%; 25-50%; >50%) and the localization of immunoreactivity were expressed for each marker. Statistical analysis was performed to determine the correlations between markers and level or thickness of melanomas. We found a good correlation between p53 expression and melanoma thickness (P <0.005), PCNA and P-gp expression. No relationship was observed between Bcl-2 expression and the different variables considered or other markers. Our data seem to indicate an unfavorable prognostic role of higher nuclear p53 expression. However, we believe that our results need to be integrated with patients' clinical follow-up and with the study of the expression of these markers in benign melanocytic lesions to gain more accurate information about their prognostic significance.
    Tumori 01/2001; 87(3):179-86. · 1.27 Impact Factor
  • S Pistolesi · M Nuti · M Castagna · R Pingitore ·
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    ABSTRACT: The B72.3, MM1.80 and 4.63 monoclonal antibodies directed against tumor-associated antigens react in human breast also with metaplastic, preneoplastic and metastatic cells, whereas they do not react with normal adult mammary tissue. The aim of our study was to point out the expression of these antigens during mammary gland development, since tumor-associated antigens are known to represent antigens of differentiation. Fifty female fetal breasts between 20 and 40 weeks of gestational age were studied. Mammary tissue was identified only in 15 cases. B72.3, MM1.80 and 4.36 monoclonal antibodies reacted with epithelial antigens and maintained the same location and intensity in the various gestational ages. Immunoreactivity was weak, cytoplasmic and widespread for the 4.36 monoclonal antibody, intense and cytoplasmatic in a large number of cells for the B72.3 monoclonal antibody, and intense and luminal for the MM1.80 monoclonal antibody. Such data further support the hypothesis that the normal process of development and differentiation can occur during tumor progression processes. Identification of these new oncofetal markers could offer a new perspective able to recognize the different phases of neoplastic progression and could be useful for prevention.
    Tumori 11/2000; 87(4):252-5. · 1.27 Impact Factor
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    ABSTRACT: The presence of lymph node metastases is the best prognostic factor for predicting relapse or survival in melanoma patients. It has been demonstrated that melanoma metastases spread through the first lymph node(s) draining the tumor (sentinel lymph node, SN) to the lymphatic system and that detection of melanoma cells in peripheral blood directly correlates with prognosis in melanoma. To identify lymph node metastases and circulating melanocytes, we developed a single-step reverse transcriptase-polymerase chain reaction assay (RT-PCR) for detection of two melanoma-specific markers: the tyrosinase gene, which encodes an enzyme associated with melanin synthesis, and melanoma antigen-related T-cells, which are present in tumor infiltrating T-lymphocytes. This method detects two tumor cells in a background of 10(7) lymphocytes. Thirty patients with stage I-IV cutaneous melanoma entered the study. Blood samples were taken preoperatively, one month after excision of the primary melanoma lesion and the SN or total lymphadenectomy, and before the start of chemotherapy and every three months thereafter in metastatic patients. SNs were collected from 22 patients, bisected and analyzed by RT-PCR and routine pathological and immunohistochemical tests. The preliminary results indicate that RT-PCR for melanoma markers is a sensitive and valuable method for the detection of micrometastases and for early diagnosis and staging of melanoma.
    Tumori 01/2000; 86(4):336-8. · 1.27 Impact Factor
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    ABSTRACT: Histologic findings after Guglielmi detachable coils endovascular embolisation have been studied in experimental aneurysms. Few reports describe histopathologic reactions to platinum coils in humans. In this report we describe gross, light microscopic pathology and scanning electron microscopy study of a ruptured basilar tip artery aneurysm in a patient who died 16 hours following coiling.
    Interventional Neuroradiology 09/1999; 5(3):257-60. · 0.78 Impact Factor
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    Rosalba Ciranni · Maura Castagna · Gino Fornaciari ·
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    ABSTRACT: Goiter is still a frequent pathological condition of the thyroid gland. Goiter consists of an enlargement of the gland caused by several physiopathologic events, the most important of which is deficient intake of iodine. A series of eighteenth-century mummies housed in the church of Santa Maria della Grazia in Comiso includes one individual showing a pathological condition, with a very large swelling in the antero-inferior region of the neck. X-rays showed small scattered radiopaque foci. Routine histology showed a large number of circular follicles immersed in fibrous tissue. Finally, immunohistochemistry revealed a strong reactivity for thyroglobulin. The data confirmed the nature of the tissue as thyroid, and the macroscopic diagnosis is that of thyroid goiter.
    American Journal of Physical Anthropology 04/1999; 108(4):427-32. DOI:10.1002/(SICI)1096-8644(199904)108:4<427::AID-AJPA4>3.0.CO;2-7 · 2.38 Impact Factor

  • Melanoma Research 06/1997; 7(Supplement 1):S120. DOI:10.1097/00008390-199706001-00418 · 2.28 Impact Factor

Publication Stats

282 Citations
152.82 Total Impact Points


  • 1988-2011
    • Università di Pisa
      • Department of Clinical and Experimental Medicine
      Pisa, Tuscany, Italy
  • 2001
    • Università degli studi di Parma
      Parma, Emilia-Romagna, Italy