Yoshio Shiina

Expression of p53, MDM2 protein and Ki-67 antigen in recurrent meningiomas.

Association of p53 gene abnormalities with tumor progression and prognosis of many neoplasms has been demonstrated, but little is known about the clinical significance of p53 abnormalities in meningiomas. The significance of p53 protein expression in recurrent meningiomas and its relationships with MDM2 protein and proliferation activity were investigated by analyzing 39 meningiomas immunohistochemically. p53 protein was expressed in 11 (35%) of 31 non-recurrent and 7 (88%) of 8 recurrent meningiomas. A high frequency of p53 expression was observed in recurrent meningiomas, which tended to have a high p53 positive index (p53 PI), indicating that p53 immunoreactivity may be a marker for predicting tumor recurrence. Four recurrent meningiomas with high p53 PIs were analyzed by the polymerase chain reaction-single strand conformation polymorphism method to detect p53 gene mutations, but none were found in exons 4–8 of this gene. Fifteen (71%) of 21 MDM2-positive and 3 (17%) of 18 MDM2-negative tumors expressed p53 protein, showing that MDM2 expression was more common in meningiomas with p53 expression. p53 immunoreactivity in the absence of mutation may indicate stabilization of the wild type through interaction with the MDM2 protein. The Ki-67/MIB-1 proliferation index (MIB-1 PI) correlated well with recurrence. The p53-positive tumors had a significantly higher mean MIB-1 PI than p53-negative tumors, suggesting that wild-type p53 inactivation by the MDM2 protein may be involved in controlling the proliferative activity in meningiomas. In conclusion, immunohistochemical examination for p53 protein as well as proliferative activity may help predict the malignant potential of tumor recurrence.

Morphometrical analysis of nucleolin immunohistochemistry in meningiomas

Abstract Nucleolin (110 kDa) is a major nucleolar protein in eukaryotic cells and one of the nucleolar organizer region (NOR)-associated proteins. We studied immunohistochemically 32 cases of meningioma, using specific antisera against nucleolin, and analyzed various nucleolin parameters, such as the number of regions and the total area of nucleolin staining per nucleus. The mean number and area of nucleolin stainings per nucleus were compared with the histological malignancy and Ki-67/MIB-1 proliferation index; the correlation with parameters of silver-stained NOR (AgNOR) was also studied. The results showed that there were statistically significant differences in the mean number and area of nucleolin stainings per nucleus between meningiomas and other two groups, atypical and anaplastic meningiomas (P < 0.05), although there was no difference between atypical and anaplastic meningiomas. The mean number and area of nucleolin stainings per nucleus were correlated with the incidence of Ki-67 positivity and AgNOR area. In view of the technical problems inherent in AgNOR staining, immunohistochemistry for nucleolin may represent a more specific and reproducible means for NOR visualization and be a promising technique for assessing cell proliferation.

Standardization of fine needle aspiration cytology of the breast -- comparison of Auto Cyto Fix and conventional smears.

In Japan, there are some problems with fine needle aspiration (FNA) cytology of the breast, such as insufficient smeared cells, air‐drying artefact and excessive erythrocytes. Liquid‐based cytology has been found to solve these problems. Equipment for such preparations has been developed, but can be expensive to purchase and operate. We developed Auto Cyto Fix 1000 (ACF), which is inexpensive and automatically smears and fixes cells. The purpose of this study was to compare the various cytological features of conventional and ACF specimens. We evaluated whether the ACF method would be able to replace the conventional method. Forty‐eight FNA specimens of breast were studied. All specimens were prepared by the direct smeared (DS) and ACF methods and evaluated for unsatisfactory cell collection, air‐drying artefacts, background findings and epithelial cell findings. Although ACF specimens were prepared using the cells remaining in the needle and syringe after preparing DS specimens, the cellularity of two of the ACF specimens was better than that of the corresponding DS specimens. ACF specimens never showed air‐drying artefact. Unlike DS specimens, which have many erythrocytes in the background, erythrocytes were filtered out and the background of ACF specimens was clean. We believe that many problems attributable to conventional FNA specimen preparation have been solved in this study. Preparation using the ACF apparatus can reduce running costs and can be used to prepare FNA specimens of the breast for cytological examination as an alternative to the conventional method.

Improved accuracy of cytodiagnosis using the Kato self-collection devise: the usefulness of smear preparation in liquid-based cytology methods.

OBJECT In the present study, we compared the positive cytodiagnostic test rates with discrepancies using self-collection devices for cervical cancer screening. We made this survey to examine whether or not our self- smear preparation method using the Kato self-collection device contributed to an improved rate of detecting atypical cells compared with existing recommended preparation methods. METHODS Specimens were collected at 14 facilities handling self-collection methods, and samples were collected by a physician in 2 facilities. The chi- squared test was performed using the SPSS ver. 20 statistical software to determine the relationships between the positive cytodiagnostic rate, specimen preparation methods, and self-collection devices. RESULTS Collecting cells using the Kato self-collection device and preparing liquid-based specimens, we obtained a significantly higher rate of positive cytodiagnosis and our results were equal to those obtained with the direct method. CONCLUSIONS Taking into consideration increased needs for screening using the self-collection method in future, with even more improved test accuracy, a screening test that is acceptable to society needs to be established.

Cytological evaluation of angiogenesis in endometrial aspirates.

Cytological evaluation of angiogenesis in endometrial aspirates

Method of producing tissue sections from endometrial scrapings

Method of producing tissue sections from endometrial scrapings

Morphological study of inclusion in Chlamydia pneumoniae infected cells.

Human line (HL) 細胞を用いChlamydia pneumoniae (C. pneumoniae) 感染細胞に出現する封入体の形態学的観察を行った.その結果, 封入体の染色性はPapanicolaou染色ではヘマトキシリン好性, Giemsa染色では好塩基性を示した.封入体は感染初期で核に隣接した一側に局在し, 感染力価の高い菌液を使用した場合は同部位に複数個観察された. さらに複数個の封入体は直径約6μmまでは融合することなしに増大し, その後融合して1つの大型封入体になった. しかし, それら封入体は部分的に核を圧排するものの, 核を完全に圧排して印環型を呈するものはなかった. 封入体周囲に膜様構造はみられず, 細胞質と封入体の間に不染間隙を認めた. 封入体の形は円形が主体であったが, 崩壊過程の所見として中心部に空洞化を示すものも観察された.以上の所見より, Papanicolaou染色およびGiemsa染色において, C. pneumoniae感染による封入体の検出は可能であるが, 宿主細胞によってはその形態が異なる可能性がある点と封入体を形成する感染細胞の出現率が低い点を考慮する必要があるように思われた.

Studies related to the significance of the occurrence of clue cells in cervico-vaginal smears.

子宮腟部擦過標本8,824例を用い, 本邦婦人におけるclue cellの出現率, 年代分布, 細胞診クラス別出現率, 好中球の量および性周期との関係について検索した.その結果, clue cellは300例 (3.4%) に出現し, 年代別には10代 (15.4%), 20代 (7.2%), 30代 (2.7%), 40代 (4.5%), 50代 (2.0%), 60歳以上 (0.3%) と加齢とともに減少する傾向を認めたが, 季節的変動はみられなかった.細胞診クラス別出現率はI (2.9%), II (4.0%), III (13.5%), IV (20.0%), V (0%) とクラスIやIIに比べIIIやIVで高く, クラスIIIで異形成と診断された28例中14例 (50.0%) にhuman papilloma virus (HPV) 感染を示唆するkoilocyteが観察された.背景に出現する好中球の量は (-) が10.3%,(1+) が68.7%,(2+) が17.3%,(3+) が3.7%で炎症像を呈す症例はわずか21%であった.性周期別には月経期 (13.1%), 増殖期 (63.7%), 分泌期 (23.2%) とホルモン環境との関係が示唆された.以上の成績より, 若年婦人やクラスIIIと診断されたHPV感染者で出現頻度が高いことから, 性行動との関係が示唆された. また, clue cellの出現は, 腟の正常細菌叢の乱れを反映したもので, その数の多少に関わらず細菌性腟症の予備群として注目すべきであり, 臨床医への報告は意義あるものと考える.