PubMed Health⌕ Search

Biomedical subjects

Stefano Calvieri

Publications and source records attributed to Stefano Calvieri.

12 recordsLinked to original sources

Management of skin ulcers in a patient with mycosis fungoides.

We present a patient with a cutaneous T-cell lymphoma/mycosis fungoides (CTCL/MF) followed for more than 10 years. After several different aggressive treatments to control progression of CTCL/MF, the patient developed several ulcerated tumors on the abdomen and limbs. Specific systemic antibiotic therapy failed to treat skin infection. While treating the stage III CTCL with polychemotherapy, we used an active colloidal hydrogel topically to manage wound healing and to treat and prevent potential sources of sepsis. After 11 weeks of treatment we observed complete cicatrization of ulcerated tumors. We reported on this case to describe the importance of a correct management of skin ulcers in immunosuppressed patients in order to avoid possible systemic spread of infection which represents the major cause of death in these patients.

Adult↗

Complete remission of brain metastases in three patients with stage IV melanoma treated with BOLD and G-CSF.

BACKGROUND: Brain metastases are the most life-threatening among the secondary localizations of melanoma for their unresponsiveness to the surgical, radiotherapeutic and/or chemotherapeutic treatments. METHODS: Accidentally, we observed a complete response (CR) in a patient undergoing chemotherapy with bleomycin, vincristine or Oncovin, CCNU or lomustine, dacarbazine (BOLD) regimen for metastatic melanoma including brain metastases, who was also treated with G-CSF to manage a concomitant leukopenia. After this observation, seven more patients with stage IV melanoma with brain metastases were treated with BOLD regimen repeated every 6 weeks with administration of G-CSF in the intervals. RESULTS: Three patients presented CR (37.5%). Two patients stopped the treatment after two courses for evident progressive disease (25%). The other three patients showed stable disease (SD: 37.5%). Median duration of SD was 24 weeks. Among the eight patients, six (75%) achieved clinical benefit. Median time to progression was 8.5 months (range 0-74+ months). Median survival was 12.5 months (range 4-74+ months). Two patients are still alive and disease-free after 74 and 57 months, respectively. CONCLUSION: We believe that the brilliant CR, the long duration of the disease-free intervals and the long survival in at least three of eight patients should encourage further research on BOLD with G-CSF for the treatment of advanced melanoma.

Adult↗

Cutaneous implantation metastasis of cholangiocarcinoma after percutaneous transhepatic biliary drainage.

Percutaneous transhepatic biliary decompression is a preoperative surgical adjunct in patients with obstructive jaundice that has been in use since 1973. It is recommended that this procedure be adopted for both palliative treatment in unresectable patients and as a preoperative means of lowering serum bilirubin in patients with potentially resectable malignancies of the pancreas or biliary tract. Metastatic tumor seeding along the transhepatic biliary catheter is an unusual complication resulting from this procedure but there have been a few cases reported in the literature. Below is a report on a 59-year-old woman in whom the percutaneous transhepatic catheter drainage of the biliary tree, performed before surgical resection of a cholangiocarcinoma, caused cutaneous tumor implantation at the catheter site 3 months later. The clinical aspect was morphea-like and histopathologic examination revealed typical features of a dermal metastasis of adenocarcinoma. Immunohistochemistry revealed cytoplasmic positivity for cytokeratin 7-19, specific for the biliary tract epithelium. A review of the literature available led us to conclude that port-site metastasis in patients with obstructive jaundice treated with percutaneous transhepatic biliary decompression was an unusual but possible complication. In fact, many catheter-tract metastatic deposits in the liver parenchyma, detected at autopsy or on operation, are mistakenly identified as hematogenous or lymphatic metastasis and are not attributed to a catheter-related process. We also report on this case because of the atypical morphea-like aspect of the skin metastasis.

Bile Duct Neoplasms↗

Fluorescence in situ hybridization analysis of allelic losses involving the long arm of chromosome 17 in NF1-associated neurofibromas.

Neurofibromatosis type 1 (NF1) is a common autosomal dominant condition associated with germline mutations of the NF1 gene located at chromosome band 17q11.2. Molecular analysis of a number of NF1-specific tumors has shown the inactivation of both NF1 alleles during tumorigenesis, supporting the tumor suppressor hypothesis for the NF1 gene. Using interphase dual-color fluorescence in situ hybridization (FISH) technique on paraffin-embedded tissues, we studied 11 plexiform, 4 cutaneous, and 6 subcutaneous neurofibromas. Cytogenetic analysis was conducted using two probes, one specific for the NF1 region (RP11-229K15) and one for the centromeric region of chromosome 17 as control. No large somatic deletions were found. Only in one of the plexiform neurofibromas loss of a whole chromosome 17 was observed. If we assume that dual-color FISH analysis is sensitive enough to detect the majority of large somatic deletions present, then other mutational mechanisms affecting the NF1 gene are probably involved in neurofibroma formation, and other tumor suppressor genes may play an important role in NF1 tumorigenesis.

Adolescent↗

Novel and recurrent mutations in the NF1 gene in Italian patients with neurofibromatosis type 1.

Neurofibromatosis type 1 (NF1) is one of the most common autosomal dominant disorders in humans, affecting 1 in 3500 individuals. NF1 is a fully penetrant exhibiting a mutation rate some 10-fold higher compared to most other disease genes. As a consequence, a high number of cases (up to 50%) are sporadic. Mutation detection is complex due to the large size of NF1 gene, the presence of pseudogenes and the great variety of lesions. In the present study we attempted to delineate the NF1 mutational spectrum in the Italian population reporting four-year experience with the direct analysis of the whole NF1 coding region in 110 unrelated subjects affected by NF1. For each patient, the whole coding sequence and all splice sites were studied for mutations, either by the protein truncation test (PTT), or, most often, by denaturing high performance liquid chromatography (DHPLC). Mutations were identified in 75 (68%) patients. Twenty-two mutations were found to be novel. The detection rate for the different methods was 7/18 (39%) for PTT, and 68/103 (66%) for DHPLC. The mutations were evenly distributed along the NF1 coding sequence. Thirty-two of the 75 unrelated NF1 patients in which germline mutations were identified (32/75, 43%) harbour 23 different recurrent mutations. Fifteen sequence variants likely to represent non-pathogenic polymorphisms were observed at the NF1 locus. Genotype-phenotype analysis was unable to detect any obvious correlation.

Chromatography, High Pressure Liquid↗

99mTc-interleukin-2 scintigraphy as a potential tool for evaluating tumor-infiltrating lymphocytes in melanoma lesions: a validation study.

UNLABELLED: Cutaneous melanoma is often characterized by the presence of tumor-infiltrating lymphocytes (TILs). The degree of such infiltration and cell activation are considered significant prognostic factors reflecting the host's immune response to the tumor; thus, patients with peritumoral infiltration may have a better prognosis and may also achieve a better response to interleukin-2 (IL2) immunotherapy. There is evidence that the expression of cluster designation (CD) 25 antigen (IL2 receptor [IL2R]) is a good marker of activity of T lymphocytes against melanoma cells. The aim of this study was to evaluate in vivo the binding of 99mTc-IL2 to lymphocytes infiltrating cutaneous melanoma and to determine whether such uptake correlates with immunologic and histologic data, thus providing useful prognostic information for IL2 therapy in patients with advanced disease. METHODS: Thirty patients with cutaneous lesions suspected of being melanoma were studied. Planar gamma-camera images over known tumor sites were acquired 1 h after the injection of 111-185 MBq of 99mTc-IL2. Tumor uptake of 99mTc-IL2 was measured as a target-to-background (T/B) radioactivity ratio. All patients underwent surgery, and histologic evaluation of the resected lesion was performed. The percentage of different peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD16, CD25) and the percentage of IL2R-positive tumor cells on histologic sections were also measured. RESULTS: At final histology, 21 lesions were found to be melanoma and 9 were classified as benign. In 15 of 21 (71%) melanomas and 2 of 9 (22%) benign cutaneous lesions, we found uptake of 99mTc-IL2. The calculated T/B ratios correlated significantly with the number of IL2R-positive TILs. CONCLUSION: 99mTc-IL2 scintigraphy provides a means of in vivo measurement of the extent of tumor infiltration of IL2R-positive cells, thereby providing valuable prognostic information for selection of patients who may benefit from IL2 immunotherapy.

Feasibility Studies↗

Survivin, bcl-2, bax, and bcl-X gene expression in sentinel lymph nodes from melanoma patients.

PURPOSE: The expression of apoptosis-related genes, such as survivin, bcl-2, bcl-X, and bax, has been evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) and by immunohistochemistry in sentinel lymph nodes (SLNs) from melanoma patients and then correlated to the outcome of patients. PATIENTS AND METHODS: Thirty-six SLNs were examined. After RNA extraction, an RT-PCR followed by Southern blot hybridization was performed to detect survivin, bcl-2, bcl-X, and bax mRNA. bcl-2, survivin, and bax gene expression was evaluated, whenever possible, also by immunohistochemistry at the protein level. RESULTS: We found a significant correlation (P <.005) between survivin expression and outcome of patients; in fact, 61.5% of patients expressing survivin gene progressed or died because of the disease, whereas 38.5% are currently disease-free. Among patients negative for survivin expression, 100% are disease-free after a median follow-up time of 52.9 months. We did not find a significant correlation between bcl-2, bax, and bcl-X gene expression and outcome of patients. In fact, these genes were found equally expressed in patients with disease progression and in disease-free patients. CONCLUSION: Our findings show a variable expression of apoptosis-related genes in SLNs of melanoma patients; more interestingly, we found that survivin expression correlates to outcome of patients in a statistically significant way, whereas the expression of other genes, such as bcl-2, bax, and bcl-X, did not seem to correlate to progression of disease. We suggest that the detection of survivin gene expression by RT-PCR in SLNs may be a useful prognostic indicator.

Adult↗

NF1 gene analysis based on DHPLC.

The high mutation rate at the NF1 locus results in a wide range of molecular abnormalities. The majority of these mutations are private and rare, generating elevated allelic diversity with a restricted number of recurrent mutations. In this study, we have assessed the efficacy of denaturing high-performance liquid chromatography (DHPLC), for detecting mutation in the NF1 gene. DHPLC is a fast and highly sensitive technique based on the detection of heteroduplexes in PCR products by ion pair reverse-phase HPLC under partially denaturing conditions. We established theoretical conditions for DHPLC analysis of all coding exons and splice junctions of the NF1 gene using the WAVEmaker software version 4.1.40 and screened for mutations a panel of 40 unrelated NF1 patients (25 sporadic and 15 familial), genetically uncharacterized. Disruptive mutations were identified in 29 individuals with an overall mutation detection rate of 72.5%. The mutations included eight deletions (exons 4b, 7, 10a, 14, 26, and 31), one insertion (exon 8), nine nonsense mutation (exons 10a, 13, 23.1, 27a, 29, 31, and 36), six missense mutations (exons 15, 16, 17, 24, and 31), four splice errors (exons 11, 14, 36, and 40) and a complex rearrangement within exon 16. Eighteen (62%) of the identified disruptive mutations are novel. Seven unclassified and three previously reported polymorphisms were also detected. None of the missense mutations identified in this study were found after screening of 150 controls. Our results suggest that DHPLC provides an accurate method for the rapid identification of NF1 mutations.

Adult↗

Prognostic significance of reverse transcriptase-polymerase chain reaction-negative sentinel nodes in malignant melanoma.

BACKGROUND: Polymerase chain reaction (PCR) is a molecular biology technique that can detect a single metastatic cell in 10(6) to 10(7) normal cells. Its use has been proposed as an additional new method for the detection of malignant melanoma nodal metastases in the sentinel lymph node (SLN) to improve the detection rate guaranteed so far by standard histology (hematoxylin and eosin; H&E) and immunohistochemistry (IHC). METHODS: Since October 1995, 137 patients with primary cutaneous melanoma (Breslow thickness,.75-4 mm) have undergone surgery for selective lymphadenectomy. To identify the SLNs, every patient had preoperative lymphoscintigraphy and a vital dye perilesional injection, followed by a gamma probe-guided operation. RESULTS: In 134 patients at least one SLN was detected, with a detection rate of 98%. Every SLN was examined by H&E and IHC (S-100 antigen and HMB-45 protein). The messenger RNA codifying for tyrosinase and MART-1 (melanoma antigen recognized by T cells) was used as the target sequence for the reverse transcriptase (RT)-PCR. The results showed 11% positive SLNs with IHC and H&E examination and 63% with RT-PCR. No recurrence was noted at follow-up in the group with RT-PCR-negative nodes (absence of false-negative cases). CONCLUSIONS: In our experience, RT-PCR SLN negativity is achieving a very favorable prognostic significance. However, RT-PCR positivity is still to be evaluated. Furthermore, results obtained with this method have been shown so far to be independent of Breslow's tumor thickness.

Antigens, Neoplasm↗