Mutation frequencies of GNAQ, GNA11, BAP1, SF3B1, EIF1AX and TERT in uveal melanoma: detection of an activating mutation in the TERT gene promoter in a single case of uveal melanoma.

BACKGROUND: Uveal melanoma is the most frequent primary tumour of the eye. It is molecularly clearly distinct from cutaneous melanoma and shows a different pattern of driver mutations. The influence of sunlight ultraviolet (UV) exposure on the aetiology of uveal melanoma is a matter of debate. The recent identification of driver mutations in the promoter of the telomerase reverse transcriptase (TERT) gene with UV-induced cytidine-to-thymidine transitions in cutaneous melanoma prompted us to investigate whether these mutations also occur in uveal melanoma. METHODS: We analysed 50 cases of uveal melanoma obtained from enucleation surgery for mutations in the genes GNAQ, GNA11, BAP1, SF3B1, EIFAX1 and TERT, measured gene expression using microarrays and analysed gene copy numbers by SNP arrays. RESULTS: We detected a TERT mutation in only one case of a 57-year-old white male patient with clinical and histopathological features typical for uveal melanoma. The tumour showed mutations in GNA11 and EIF1AX that are typical for uveal melanoma and absent from cutaneous melanoma. No mutations were detected in GNAQ, BAP1 and SF3B1 that are frequently mutated in uveal melanoma. Both copies of chromosome 3 were retained. Several tumours among which the one carrying the TERT promoter mutation showed elevated TERT expression. Consistent with previous reports, GNAQ is inversely associated with chromosome 3 monosomy and metastasis. BAP1 mutations are significantly associated with chromosome 3 monosomy but not with relapse. CONCLUSION: These data indicate that TERT mutations are rare in uveal melanoma. No conclusion can be drawn on their potential influence on tumour progression.

Second-line treatment with intravenous gemcitabine and oral etoposide in platinum-resistant advanced ovarian cancer patients: results of a phase II study.

OBJECTIVE: The outcome of advanced ovarian cancer patients has not significantly improved since the introduction of platinum. One of the major reasons for this failure is the lack of an effective second-line treatment. In this phase II trial we tested the combination of gemcitabine and etoposide in 2 different groups of patients. Group 1 consisted of patients showing disease progression or relapse within 6 months of first-line platinum-based chemotherapy. Group 2 comprised heavily pretreated patients showing progression during the last chemotherapy attempt. METHODS: Thirty-four patients were enrolled. Gemcitabine was administered at a dose of 1,000 mg/m(2) on days 1 and 8 and etoposide was administered orally at 100 mg/day on days 8-12 for 6 courses. RESULTS: Eighteen patients (52.9%) had an objective response and the median duration of the response was 10.3 months. Our chemotherapy regimen showed a low toxicity and good patient compliance. In 5 patients the treatment had to be delayed and in only 2 patients it was discontinued. CONCLUSIONS: The combination of gemcitabine and oral etoposide seems to be a safe and effective second-line treatment for platinum-resistant ovarian cancer patients. Additional data on larger series are warranted to better define the activity of this combination regimen.

Placental lesions in a case of DiGeorge sequence.

This work describes some placental alterations found in a partial form of DiGeorge sequence, namely, hypoplasia of a cord artery with internal calcification of an extensive endoluminal thrombosis, and widespread calcification of microthrombi in the arteries of the second and third order villous branches. Hypoplasia of a cord artery is a relatively rare event, and is also associated with malformations of the gastroenteric and cardiovascular system, as sometimes described in the DiGeorge sequence. Interesting placental alterations are reported and their likely physiopathologic basis and pathogenic correlation discussed in order to give a better and more comprehensive picture of the DiGeorge sequence in which the correlated placental alterations are not sufficiently known.

[Applications and advantages of a multimedia system for autopsies ]. / Applicazioni e vantaggi dei sistemi multimediali nella pratica autoptica.

This work evaluates the benefits and applications of computers and multimedia systems in post-mortem examination practice and, more in particular, in the definition of data collection protocols. We examined issues concerning the different aims of autopsy (e.g. diagnostic, scientific, educational, legal), and found that the pathologist's main duty is to acquire a large amount of data in the best possible way. However, despite the will to carry out detailed post-mortem examinations, many pathologic anatomy services face objective difficulties in doing so, especially due to understaffing, lack of time and high costs. The Institute for Pathologic Anatomy of the University of Genoa has developed software for data handling and for outcome reporting, a particularly important aspect in fetal-perinatal diagnosis. The system consists of a relational database in a client-server environment (Fourth Dimension) with two integrated parts. The first part, with unrestricted access, contains patients' personal data, including gender, age, time and date of death, hospital department of origin, person and department requiring the post-mortem examination, hour and time of autopsy, pathologist's name, and clinical diagnosis of death. Using a scanner, a copy of the autopsy application is also field, together with the patient's medical file and any diagnostic images useful to document the case history. The second part of the information system is accessible by pathologists only, and contains the autopsy report. This part is organized to balance two different needs: it allows sufficient space and freedom for autopsy description while providing guidelines for presentation of the report. The structure of the conventional autopsy protocol has been maintained, with subdivisions for all the organs and apparatuses according to topographic criteria. Before this part, a section is dedicated to external cadaver examination and anthropometric data; weight, shape, volume and texture are described for each organ, together with external and cut-surface features. A third section allows the examiner to report other observations not requested previously, while a final section is also provided for the epicrisis and for the formulation of the final diagnosis, the same as that reported in the first form. The database is coupled with an interactive system for collecting voice comments, thereby replacing the need for tape-recorders in the autopsy room. The user can recall a dictation window, dictate a text, check spelling and insert additional text. The database is also coupled to an image acquisition system, on the assumption that moving images allow a more faithful documentation of reality. Therefore, all rooms in which autopsies are carried out on fetuses or neonates have been equipped with a fixed camera linked to a monitor and a video-recorder. A PCB, used for image digitalization, recognizes up to 16,000,000 different colors. Guided by dedicated software, image files are transferred to a computer and then saved with the autoptic report. The database can be consulted and queried in two principle ways: by key words in the contents or main disease descriptions, or by individual words or phrases contained within the complete text of the reports. The present database system for autopsy reporting has proved itself useful in a pathological anatomy service. The combined presence of images and texts renders the system useful also as a research tool. By linking to a Web site dedicated to pathologic anatomy, it will be possible to display online rare cases involving diagnostic difficulties. The system offers great advantages for present and retrospective diagnostics, as well as for research and education purposes.

Standardized AgNOR analysis as a prognostic parameter in endometrial carcinoma, endometrioid type.

OBJECTIVE: To evaluate interobserver reproducibility of histologic grade in endometrial adenocarcinomas of endometrioid type (EC), to assess the relationships between nuclear grade and the amount of argyrophilic nucleolar organizer region (AgNOR) proteins and to determine the prognostic value of AgNOR proteins and the main clinicopathologic parameters. STUDY DESIGN: Architectural and nuclear grading were independently assessed by two pathologists in 64 formalin-fixed, paraffin-embedded surgical samples of EC obtained from an equal number of patients (age range, 38-84 years; mean, 63.5). Interobserver agreement was determined using the kappa statistic; discrepant cases were reviewed, and a consensus was reached. Standardized AgNOR analysis was performed according to the guidelines of the Committee on AgNOR Quantification, measuring the mean area of AgNORs per nucleus (NORA) by an image analysis system. RESULTS: The kappa values for interobserver agreement were substantial for architectural grading and moderate for nuclear grading. When NORA values were compared to the nuclear grade assessed by different observers, the most significant linear correlation (r = .713, P < .001) was found for the nuclear assessment obtained by consensus of the two pathologists. Moreover, statistical analysis allowed discrimination of architectural grade 1 from grade 2 and 3 EC. By the Kaplan-Meier method, the prognosis was worse for patients with higher NORA values (> 4.212 micron 2), while, by Cox multivariate analysis, AgNOR quantity emerged as an independent prognostic variable. CONCLUSION: Use of standardized AgNOR analysis may be an additional and objective tool in the assessment of histologic grade as well as a reliable method of determining prognosis in EC.

Morphologic and biologic studies on ten cases of verrucous carcinoma of the vulva supporting the theory of a discrete clinico-pathologic entity.

Ten cases of verrucous carcinoma (VC) of the vulva diagnosed from January 1989 to December 1996 were studied. Patient age ranged from 50 to 83 years. The following examinations were performed on buffered formalin-fixed material: 1). in situ DNA hybridization, probes HPV 6/11, 16/18, 31/35/51; and 2). a series of immunohistochemical stainings to demonstrate wild and mutant types of the p53 protein, cytokeratin expression and pattern distribution (AE1 and AE3), and proliferating pattern (MIB 1). In situ DNA hybridization analysis for human papillomavirus 6/11, 16/18, 31/35/51 was negative in all cases. Wild and mutant types of p53 protein transcribed from related oncosuppressor gene were not detected. Keratins AE1 and AE3 showed a peculiar distribution pattern, that is, AE1 was uniformly positive in the surface and intermediate layers, while it was almost negative in the basal layer which-on the contrary-was mainly positive to AE3 keratins. MIB-1 highlighted 10-40% of proliferating cells; however, in all cases, 70-80% of MIB-1 positivity was found in the basal layer of the neoplastic epithelium. These results seem to show the morphofunctional and growth characteristics of neoplastic epithelium, thus stressing that VC should be considered as a discrete entity in vulvar tumors.