Shinkichi Sato

Cellular localization of insulin-degrading enzyme in rat liver using monoclonal antibodies specific for this enzyme

Although insulin-degrading enzyme (IDE) has been implicated in the intracellular degradation of insulin, the cellular localization of this enzyme is still controversial. In the present study, we have examined the cellular localization of IDE in the rat liver by three different techniques using monoclonal antibodies. First, direct immunohistochemical staining of rat liver with one of the monoclonal antibodies revealed that IDE immunoreactivity mainly exists in parenchymal cells, especially in the vicinity of the portal tract and also in the epithelium of the bile duct under light microscopy. In the electron microscopic study, IDE immunoreactivity was found in the cytoplasm near the rough endoplasmic reticulum but not in the plasma membrane, nucleus, or mitochondria. Second, immunoblotting analysis of the subcellular fraction in rat liver showed that the monoclonal antibody specifically reacted with a single polypeptide in the cytosolic fraction, of apparent Mr 110,000, which was consistent with the Mr of IDE. However, a polypeptide band corresponding to IDE could not be observed in the plasma membrane, mitochondrial, or lysosomal fraction. Third, IDE was only detectable in the cytosolic fraction by sandwich radioimmunoassay using two monoclonal antibodies. These results all suggest that IDE is a cytosolic enzyme.

Visualization, quantification and therapeutic evaluation of angiogenic vessels in cancer by synchrotron microangiography

The usefulness of a synchrotron microangiography system for depicting, quantitating and therapeutically evaluating angiogenic vessels in cancer is illustrated. In 20 mice transplanted with murine colon cancer, sequential changes in the angiogenic vessels were determined by using synchrotron microangiography, using changes in tumor volume for reference. This system allowed the depiction and quantification of angiogenic vessels in the period from one to four weeks after transplantation. The effects of antiangiogenic therapy were evaluated by using a neutralizing antibody against vascular endothelial growth factor. The neutralizing antibody partially suppressed angiogenesis and tumor growth. Synchrotron microangiography is shown to be useful for the depiction, quantification and evaluation of angiogenic vessels in cancer.

Gallbladder adenocarcinoma with human chorionic gonadotropin: a case report and review of literature

BackgroundThe case of adenocarcinoma with human chorionic gonadtropin (HCG), primary in the male gallbladder, is extremely rare. A Medline search has shown only a few similar cases reported.MethodsWe herein describe a case of primary gallbladder adenocarcinoma associated by ectopic HCG positive tumor cells in a 79-year-old male.ResultsPathological examination showed a mixture of moderately and poorly differentiated adenocarcinoma with ectopic HCG and placental alkaline phosphatase (PlAP) in tumor cells, though the increase of serum or urinary HCG secretion was not confirmed. The literatures were also reviewed.ConclusionsA case of gallbladder cancer with ectopic HCG production is quite rare in the literature, though many similar cases in other site, especially in GI tract, are reported. Embryological consideration suggests the increased frequency of similar cases more than being thought now.

Sensitivity to CPT-11 and platinum derivatives of stage III/Dukes' C colorectal cancer with occult neoplastic cells in lymph node sinuses

Among 13 patients with recurrent colorectal cancer (recurrence group: RG), immunohistochemical expression of Topo-1 was high in 4 patients (30.8%) and low in 9 patients (69.2%), while the non-recurrence group (N-RG) (n=8) showed high expression in 1 patient (12.5%) and low expression in 7 patients (87.5%) (NS). Regarding immunohistochemical expression of Bax/ERCC-1, high Bax/low ERCC-1 expression was observed in 6 patients (46.2%) from the RG and other patterns of expression were seen in 7 patients (53.8%), while high Bax/low ERCC-1 expression was observed in 4 patients (50.0%) from the N-RG and other patterns were found in 4 patients (50.0%) (NS). PCR analysis of Topo-1 expression revealed high expression in 9 patients (75.0%) from the RG (n=12) and low expression in 3 patients (25.0%), while the N-RG (n=8) showed high expression in all 8 patients (100.0%) and low expression in none (NS). With respect to ERCC-1, PCR analysis revealed high expression in 7 patients (58.3%) from the RG (n=12) and low expression in 5 patients (41.7%), while the N-RG (n=8) showed high expression in 1 patient (12.5%) and low expression in 7 patients (87.5%) (p<0.05). These results suggest that tumor sensitivity to CPT-11 and platinum derivatives is similar in stage II colorectal cancer patients with ONCs.

Predicting the recurrence/metastasis of stage I and II breast cancer without lymph node metastasis.

This study compared prediction of the recurrence of breast cancer by detection of occult neoplastic cells (ONCs) in lymph nodes or by using the criteria for a high risk of recurrence and metastasis of gastric/large bowel cancer. The subjects were 45 patients with stage II and III node-positive breast cancer. Prediction of recurrence by detection of ONCs showed a sensitivity of 78.6% (11/14), a false-negative rate of 21.4% (3/14), a specificity of 96.4% (30/31), a false-positive rate of 3.2% (1/31), and an accuracy of 87.7% in patients with stage II and III node-positive cancer. Prediction of recurrence based on positivity for at least 2 of the high-risk criteria showed a sensitivity of 92.9% (13/14), a false-negative rate of 7.1% (1/14), a specificity of 87.1% (27/31), a false-positive rate of 12.9% (4/31), and an accuracy of 90.0% in patients with stage II and III node-positive cancer. These results suggest that ONCs plus the high-risk criteria are useful for predicting recurrence/metastasis of stage II and III node-positive breast cancer during the early postoperative period with a high sensitivity and accuracy.

Use of epithelial‐specific antigen for cytological diagnosis of glandular lesions in the uterine cervix

To the Editor: It has been reported that endocervical adenocarcinoma accounts for 5–20% of all cervical carcinomas and its incidence has been increasing in recent years. Endocervical adenocarcinoma displays less radiosensitivity than squamous cell carcinoma and effective chemotherapy has not been established. Therefore, endocervical adenocarcinoma currently has a poor prognosis, although it is hoped that early detection and early treatment may result in prognostic improvement. However, the accuracy of cytological diagnosis for detecting endocervical adenocarcinoma is not always high due to problems with the cellular sampling method and with screening. Regarding cellular sampling, if the site from which the endocervical lesion cannot be observed by colposcopy, it can be difficult to obtain sufficient amounts of tumor cells. Regarding problems with screening, concomitant squamous epithelial lesions, observed in 40–60% of patients with endocervical adenocarcinoma, making it difficult to screening for adenocarcinoma cells. Additionally, the false-negative rate of cytological diagnosis for endocervical adenocarcinoma is reported to range from 12.8% to 55.0%. This data suggests that it is difficult to use cytological diagnosis for endocervical glandular lesion. It has been reported that the expression rate of epithelialspecific antigen (ESA) is high in histologically detected endocervical adenocarcinoma. The present study was performed to determine whether or not expression of ESA is specific for endocervical lesions in cytology specimens and histological specimens. The subjects of this study were a total of 61 patients, who underwent both cytological and histological diagnosis (biopsy and excised specimens) almost simultaneously at our hospital from 1990 to 2007. They included 20 patients with endocervical adenocarcinoma (EA), 10 patients with adenocarcinoma in situ (AIS), 11 patients with carcinoma in situ (CIS), and 20 patients with invasive squamous cell carcinoma (SCC). Epithelial-specific antigen (VU1D9, 1:100; Novocastra Laboratories, Newcastle upon Tyne, UK) was detected by the indirect immunohistochemical method (MAX-PO MALT; Nichirei Biosciences, Tokyo, Japan) using HE-stained and Papanicolaou-stained (Pap-stained) and decolorized specimens. When histological specimens were investigated, antigen activation was performed by pretreatment with 0.1% trypsin at 37°C for 30 min. In both histology specimens and cytology specimens, endogenous peroxidases were inactivated by incubation with 3% H2O2 for 30 min. Both the primary antibody and the secondary antibody were reacted with the specimens for 30 min at room temperature, after which color was developed with DAB. The percentage of ESA-positive cells among 500 tumor cells was calculated in both histology specimens and cytology specimens and was classified as follows: 0% = − ; ≤ 30% = 1+ ; 30–70% = 2+ ; ≥ 70% = 3+ . Evaluation of the ESA expression showed the positive reaction to the cell membrane or cytoplasm of epithelial cells. For statistical analysis, Student’s t-test was employed and P < 0.05 was considered to be statistically significant. Expression of ESA for histological findings, normal squamous cells were generally negative for ESA, although some of the basal cells were positive. Endocervical cells were mainly negative for ESA, though the basolateral membranes were partially positive (Fig. 1a). Squamous metaplasia partially showed weak expression of ESA. Expression of ESA in endocervical lesions was seen in 85.0% of EA lesions (17/20 patients) (Fig. 1b); 100.0% of AIS lesions (10/10 patients) (Fig. 1c); 18.2% of CIS lesions (2/11 patients); and 45.0% of SCC lesions (9/20 patients). Thus, expression of ESA was detected in all patients with AIS and nearly all patients with EA (sensitivity of 90.0% and specificity of 64.5%). The level of ESA expression in patients with EA was mainly 2+ (1+ in 25.0% of the patients; 2+ in 45.0%; and 3+ in 15.0%), although there were individual differences. All of the patients with AIS had ESA-positive tumors and strong expression was frequent (3+ in 70%). On the other hand, most of the patients with CIS had ESA-negative tumors (81.8%) (Fig. 1d), but 40% of the patients with SCC had ESA-positive tumors (1+ in 25.0% and 2+ in 15.0%). In cytological specimens, normal squamous cells, squamous metaplastic cells and endocervical cells were negative for ESA or slight cytoplasmic expression in endocervical cells (Fig. 2a). Expression of ESA in endocervical lesion was seen in 100.0% of EA lesions (20/20 patients) (Fig. 2b) and 100.0% of AIS lesions (10/10 patients) (Fig. 2c) while ESA was only positive in 18.2% of CIS lesions (2/11 patients) (Fig. 2d) and 30.0% of SCC lesions (6/20 patients). Thus, all of the patients with EA or AIS were ESApositive, whereas the ESA expression rate was low in squamous lesions (sensitivity of 100.0% and specificity of 74.2%). The level of ESA expression was strong in the patients with EA or AIS (3+ in 100.0% of the patients with EA and 80.0% of the patients with AIS) (Fig. 2a, b). On the other hand, weak ESA expression was seen in patients with CIS or SCC (1+ in doi:10.1111/pin.12379 bs_bs_banner

A Case of Budd-Chiari Syndrome Associated with Polycythemia Vera.

症例は43歳, 女性. 1993年9月頃より腹水の貯留, 黄疸が出現し, 1994年1月入院. 血液検査では全血球系の著増とともに著明な肝障害が認められた. 血液所見および骨髄生検より真性多曲症ならびに急性肝不全と診断した. また腹部超音波検査にて右・中・左肝静脈は血栓で充満し面流は認められず, 血栓による肝静脈の閉塞が考えられた. 入院後急速に肝不全が進行し, 1カ月の経過でDIC, 多臓器不全のため死亡. 病理解剖では肝静脈内に器質化した血栓が認められ, 肝細胞はzone 3を中心に広範な変性, 壊死を示していた. 本邦におけるBudd-Chiari症候群の多くは膜様閉塞に伴うものであり, 肝静脈血栓によるBudd-Chiari症候群の報告は少ない. 特に真性多血症に伴う肝静脈閉塞は本例を含め2例のみで極めて稀な症例と考えられる.