Eikichi Okada

Overexpression of phosphorylated histone H3 is an indicator of poor prognosis in gastric adenocarcinoma patients.

Ki-67 immunostaining is commonly used for assessing cell proliferation, but studies of its use as a prognostic indicator have revealed discordant results in gastric cancer patients. Recently, antibodies for phosphorylated histone H3 have been used to identify dividing cells because of its precise overexpression in mitosis. The authors tested the hypothesis that phosphorylated histone H3 overexpression might be a good prognostic indicator for gastric cancer patients by conducting an immunohistochemical comparison with Ki-67 in gastric cancer samples. One hundred twenty-two surgically resected primary cases were selected and histologically categorized in accordance with Laurens classification. No correlation was found between phosphorylated histone H3 and Ki-67 regarding overexpression. However, correlations between phosphorylated histone H3 overexpression and clinicopathologic variables were noted for histologic type (intestinal type predominant in high labeling indices [LIs], defined as over the value of the 75th percentile; P<0.01), vessel invasion (positive in high LIs; P=0.05), and lymph node metastasis (positive in high LIs; P=0.04). With regard to Ki-67 overexpression, no correlation was evident with the clinicopathologic variables except histologic type (intestinal type predominant; P=0.05). By the Kaplan-Meier method with the log-rank test, cases overexpressing phosphorylated histone H3 showed a poorer prognosis than cases with low expression (P<0.01). In contrast, Ki-67 expression did not influence prognosis. Multivariate analyses indicated phosphorylated histone H3 overexpression to be an independent prognostic factor, together with lymphatic invasion and venous invasion (P<0.01). In conclusion, it seems likely that phosphorylated histone H3 plays an important role in the prognosis of gastric cancer, and its immunohistochemical investigation is useful for the prediction of prognosis in gastric cancer.

Modified and simplified western blotting protocol: Use of intermittent microwave irradiation (IMWI) and 5% skim milk to improve binding specificity

A modified western blotting protocol was developed to increase the binding specificity of antigens and antibodies, using intermittent microwave irradiation (IMWI) with seven antibodies and two cell lines. The method was based on IMWI of the blotting membrane in the immunoblotting step using 5% skim milk as the diluting buffer. For some antibodies against p53, CDK4 and cyclinE, there were no distinct differences between the IMWI(+) and IMWI(–) counterparts; but improvement over the standard protocol was noted in both. For some antibodies, such as the polyclonal antibody against tubulin and the monoclonal antibodies against β‐tubulin, cyclinA and cyclinB1 (which were otherwise difficult to obtain good results with), IMWI was extremely effective, resulting in clear, specifically binding bands and a clean background. Moreover, the times were reduced from 8 to 3 h. Both the IMWI(+) and IMWI(–) protocols can be applied as simple, rapid and highly specific detection techniques for applications with various antigens, reducing background ‘noise’ to a minimum.

CLASSIFICATION OF CARDIAC RUPTURE COMPLICATED IN MYOCARDIAL INFARCTION

Thirty of 32 cases with cardiac rupture (CR) complications in the myocardial infarction were found out of a total of 91 cases of acute myocardial infarction (AMI). The mean age of the ruptured group in females was significantly younger than that of the non‐ruptured group. Twenty‐one cases showed free wall rupture of the left ventricle, six perforation of ventricular septum and three double rupture. All cardiac ruptures occurred in cases of transmural infarction. The age of AMI was histologically estimated. Nine cases complicated within the first 24 hours of AMI showed rupture of the left anterior wall. CR in the periphery within the infarct occurred at any time during the first week after onset, and cases of the central rupture were increased in number after the 3rd day of AMI. Pathologic findings indicated that elevated wall tension was considered to be most closely related to the cause of CR. Thirty two cases of CR were classified into three types: (1) blowout type, (2) hemorrhagic dissecting type, and (3) thinning‐with‐rupture type. Hemorrhagic dissecting type was characterized by multiple endocardial ulcers and Assure canals extending from the ulcer with hemorrhage in the surrounding myocardium. Complex Assure was seen in two cases of this type.

Progeria with cardiac hypertrophy and review of 12 autopsy cases in the literature

An autopsy of an 11 years 5 months old boy with progeria is presented. Hyaluronuria, low growth rate of skin fibroblasts, decreased number of T‐cells in lymphocyte subpopulation, increase of anti DNA antibody, and antibody for microsome of thyroid gland were detected clinically. He had suffered an attack of cerebral infarct and died of congestive cardiac failure. Pathologic findings were scleroderma‐like skin atrophy, moderate arteriosclerosis of the aorta and great arteries, severely narrowed coronary sclerosis with an extensive subendo‐cardial fibrosis, a large left cerebral infarct with pin‐holed stenosis of both internal carotid arteries, cortical atrophy of thymus, and atrophic lymph nodes. The cardiac muscle fibers were slightly hypertrophied and measured 15.5 ± 2.8 m̈ in diameter. Histologic findings suggest that increasing of collagen in the connective tissue may play an important role in progeria. Further study of metabolic disturbance in the connective tissue of progeria is necessary.

High labeling indices of cdc25B is linked to progression of gastric cancers and associated with a poor prognosis.

To clarify the significance of cdc25B, which plays an important physiologic role in regulation of the G2/M check point, in progression of gastric cancer, 125 samples of paraffin-embedded gastric cancers were investigated by immunohistochemistry. In addition, 3 human gastric cancer cell lines were studied to determine the cellular localization of cdc25B by immunohistochemistry and cell fractionation followed by Western blotting. In the cell lines cdc25B was found to be present in both nuclei and cytoplasm, but predominantly in nuclei. High labeling indices of cdc25B in invasion front of gastric cancer was observed in 31 out of 125 cases (24.8%), linked to an advanced depth of cancer invasion (P=0.02), high rates of lymphatic invasion (P=0.03), and lymph node metastasis (P<0.01). Furthermore, the Kaplan-Meier method demonstrated a poor prognosis for cdc25B high labeling indices cases (P=0.02), although multivariate analysis revealed it not to be an independent factor. In conclusion, it seems likely that cdc25B is located predominantly in nuclei when overexpressed and this has some linkage with progression of gastric cancer.

CONGENITAL DILATATION OF INTRAHEPATIC BILE DUCTS ASSOCIATED WITH HEPATIC FIBROSIS and MEDULLARY SPONGE KIDNEY —RECONSTRUCTION and MICRODISSECTION STUDY

An autopsy case of a four‐year‐old female with congenital dilatation of intrahepatic bile ducts with bilateral medullary sponge kidney is presented. The liver and kidney were investigated by reconstruction with serial sections and the kidney was also studied by microdissection.

Hepatocellular carcinoma and type 2 diabetes mellitus: cytokeratin 8/18 expression in hepatocellular carcinoma and glycogen-storing hepatocytes

Cytokeratins (CKs), the intermediate filament (IF) proteins of epithelia, are sub-divided into type I (CK9-20) and type II (CK1-8) and expressed as type I/II pairs in a cell differentiation manner. In adult liver, hepatocyte IF comprise only CK8/18.[3] CK8/18 expression in normal and diseased liver has been reported, including positive expression in alcoholic steatohepatitis (ASH) and/or non-alcoholic steatohepatitis (NASH) and HCC.[3]

Hepatocellular carcinoma and type 2 diabetes mellitus: two cases highlighting changes in tumor glycogen content

This article reports two patients with hepatocellular carcinoma (HCC) and type 2 diabetes mellitus (T2DM), who showed marked changes in hepatocellular glycogen content. Periodic acid-Schiff (PAS)-positive and diastase-PAS-negative (glycogen-storing) hepatocytes were detected in both background liver parenchyma and in HCC tissues. In HCC tissues, the number of glycogen-storing cells resembling hepatocytes was considerably reduced and unevenly distributed as compared with hepatocytes in background liver. To be known, changes in hepatocellular glycogen content in T2DM patients have not been previously described. It is hypothesized that the reduction in glycogen content in both patients was likely associated with the emergence of Warburg type of glycolysis.