Daisuke Ozaki

Diagnostic Usefulness of p16/CDKN2A FISH in Distinguishing Between Sarcomatoid Mesothelioma and Fibrous Pleuritis

The distinction between sarcomatoid mesothelioma and fibrous pleuritis is difficult based on histology, especially when the amount of tumor tissue examined via biopsy is small and immunohistochemical examination is inconclusive. We studied the usefulness of deletion of p16 with fluorescence in situ hybridization (FISH) and p16 hypermethylation with polymerase chain reaction for the diagnosis and prognosis of malignant pleural mesothelioma (MPM). We analyzed 50 MPMs, including 22 sarcomatoid mesothelioma cases and 10 fibrous pleuritis cases. We set the cutoff value of homozygous deletion pattern as 14.4% based on FISH signaling patterns using samples of fibrous pleuritis. The percentage of homozygous deletion pattern was higher than 14.4% in 55.6% of the epithelioid mesotheliomas (10/18) and in all of the sarcomatoid mesotheliomas (22/22). Methylation of p16 was observed in 7 (20.6%) of 34 informative cases. p16 FISH analysis can be a reliable test for distinguishing between sarcomatoid mesothelioma and fibrous pleuritis and a prognostic factor for MPM.

Mucoepidermoid carcinoma of the thymus

Mucoepidermoid carcinoma of the thymus is a rare carcinoma and there is little agreement about the treatment of this tumor. According to the analysis of previously reported tumors, biologic behavior of the tumor correlated with the spread of the lesion and degree of differentiation. We report a case of this tumor in a 31-year-old man. Resection of the tumor included the left upper lobe of the lung, the phrenic nerve, pericardium and disseminations in the pleura. The clinicopathological feature of this case was high-stage disease and low-grade histology. Postoperative chemotherapy and radiotherapy were performed, and the patient is alive without recurrence 14 months after surgery.

Comparative morphometric studies of benign and malignant intraductal proliferative lesions of the breast by computerized image analysis

Using computerized image analysis, we newly devised a method for automatic quantitative evaluation of nuclear arrangements in variable proliferative intraductal (cribriform) lesions of the breast, by calculating angles of longest nuclear diameter to a horizon (LNDA). Internuclear correlation in the cribriform lesions was thus examined, comparing 22 benign and 62 malignant cases. In the malignant lesions, nuclear arrangements tended to be multidirectional, probably because of vertical nuclear arrangements toward acinar lumens as reflected by the wide distribution of LNDAs. Conversely, in the benign lesions, groups of nuclei tended to arrange toward one certain direction, forming a complex streaming pattern with the distribution of LNDAs usually producing a distinctive peak. Other conventional nuclear features, such as nuclear area, nuclear perimeter, shortest nuclear diameter (SND), and leading variables were also quantitatively assessed for multivariate analysis. It was noted that nuclear arrangements could represent a favorable discriminator. Linear discriminant function could classify 66 of the total 84 cases (78.6%) as being consistent with the diagnosis of pathologists. We conclude that internuclear arrangement could be a useful discriminating variable in benign and malignant breast lesions for the further development of an automatic analytic system.

Cytologic Differential Diagnosis of Malignant Mesothelioma and Reactive Mesothelial Cells With FISH Analysis of p16

Mesothelioma patients often present with serosal effusions, which are ideal for cytopathological diagnoses. However, the morphological overlap between malignant and benign mesothelial proliferation can make a conclusive cytological diagnosis of mesothelioma elusive because immunohistochemical staining does not discriminate definitively between the two in this setting. p16 is deleted in up to 80% of pleural mesotheliomas. The aim of this study was to establish the correlation between the p16 deletion status of the cell block with that of its corresponding tumor using fluorescence in situ hybridization (FISH) analysis for individual patient tumors.

Interstitial invasion of well-differentiated hepatocellular carcinoma and subsequent tumor growth.

Pathological processes of interstitial invasion of hepatocellular carcinoma (HCC) were investigated in 125 autopsy and seven surgical cases, where selection focused on those occurring in well‐differentiated HCC. The invasive pattern of well‐differentiated HCC was characterized by the formation of streaks of tumor cell cords running along connective tissue fibers, referred to as a streak pattern. These cell cords were unaccompanied by reticulin frameworks and type IV collagen, which are consistent components of HCC parenchyma. The typical streak pattern was observed only in HCC cases and could be useful in differential diagnosis. Following progressive accumulation or proliferation of the tumor cells within the interstitium, many capillaries appeared between the cell cords while collagen fibers tended to disappear. The tumor cell cords then showed branching and were accompanied by reticulin frameworks, type IV collagen and sinusoidal blood spaces. In the setting of interstitial invasion of well‐differentiated HCC, it is suggested that interstitial tissue is converted into HCC parenchyma via the aforementioned steps.

Characterization of hyperplastic foci observed in surgical specimens of hepatocellular carcinoma

By reviewing previous surgical specimens of hepatocellular carcinoma, 17 cases with hyperplastic foci (HPF) characterized by discernible increase in nuclear densities, could be histologically selected. Nuclear densities of HPF and control hepatic parenchyma were assessed quantitatively by counting the nuclear number of hepatic cells, and proliferating cell nuclear antigen labeling index was measured. HPF occurred multifocally, confined within a lobular unit, smoothly merging into surrounding hepatic parenchyma. Nuclear densities of HPF were 1.71 times greater than those of control hepatic parenchyma. The hepatocytes of HPF also showed significantly higher proliferative activities than those of control parenchyma. In addition, noticeable structural distortions, such as focal trabecular thickening or microacinar formation of hepatocytes, were sometimes observed in HPF. However, these HPF seemed to be distinguished from minute de novo hepatocellular carcinoma (HCC) or intrahepatic HCC metastasis, because of paucity of distinctive atypical changes, and intimate correlation with neighboring hepatocytes. Several adjacent HPF were aggregated to form a much larger unit of a hyperplastic area with loss of fibrous septa of liver cirrhosis. It was suggested that grossly detectable large regenerative nodules are produced via fusion of several adjacent HPF.

Usefulness of p16/CDKN2A fluorescence in situ hybridization and BAP1 immunohistochemistry for the diagnosis of biphasic mesothelioma ☆

Malignant mesothelioma is a highly aggressive neoplasm, and the histologic subtype is one of the most reliable prognostic factors. Some biphasic mesotheliomas are difficult to distinguish from epithelioid mesotheliomas with atypical fibrous stroma. The aim of this study was to analyze p16/CDKN2A deletions in mesotheliomas by fluorescence in situ hybridization (FISH) and BAP1 immunohistochemistry to evaluate their potential role in the diagnosis of biphasic mesothelioma. We collected 38 cases of pleural mesotheliomas. The results of this study clearly distinguished 29 cases of biphasic mesothelioma from 9 cases of epithelioid mesothelioma. The proportion of biphasic mesotheliomas with homozygous deletions of p16/CDKN2A in total was 96.6% (28/29). Homozygous deletion of p16/CDKN2A was observed in 18 (94.7%) of 19 biphasic mesotheliomas with 100% concordance of the p16/CDKN2A deletion status between the epithelioid and sarcomatoid components in each case. Homozygous deletion of the p16/CDKN2A was observed in 7 (77.8%) of 9 epithelioid mesotheliomas but not in fibrous stroma. BAP1 loss was observed in 5 (38.5%) of 13 biphasic mesotheliomas and in both epithelioid and sarcomatoid components. BAP1 loss was observed in 5 (62.5%) of 8 epithelioid mesotheliomas but not in fibrous stroma. Homozygous deletion of p16/CDKN2A is common in biphasic mesotheliomas, and the analysis of only one component of mesothelioma is sufficient to show that the tumor is malignant. However, compared with histology alone, FISH analysis of the p16/CDKN2A status and BAP1 immunohistochemistry in the spindled mesothelium provide a more objective means to differentiate between biphasic mesothelioma and epithelioid mesothelioma with atypical stromal cells.

Malignant pleural mesothelioma: Clinicopathology of 16 extrapleural pneumonectomy patients with special reference to early stage features

The earliest pathological events in the development of malignant pleural mesothelioma (MPM) are not understood. The aim of the present study was to elucidate the early histopathological features of MPM. A total of 16 extrapleural MPM pneumonectomy patients were investigated. Early stage mesothelioma was arbitrarily defined as a tumor ≤5 mm in thickness regardless of the nodal status or other organ involvement. Eight of these patients (six with epithelioid, two with biphasic) had early stage mesothelioma by this definition. Macroscopically there was no visible tumor, but the parietal and visceral pleura were thickened and there was focal adhesion between them. Microscopically, the mesothelioma lesions were multifocal and discontinuous on the pleura. In extremely early cases of epithelioid mesothelioma, tumor cells were generally arrayed in a single layer, but papillary proliferation was observed elsewhere. In sarcomatoid mesothelioma, mesothelioma cells proliferated, forming multiple small polypoid nodules on the pleura. Epithelial membrane antigen was helpful to distinguish reactive from neoplastic mesothelium, but glucose transporter‐1 was not. Mesothelioma cells disseminate diffusely throughout the parietal and visceral pleura and mediastinal fat tissue before becoming visible. Stage Ia mesothelioma (neoplasm limited to the parietal pleura) would not be observed in daily practice.