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Objective Nonobstructive general angiography (NOGA) is a novel modality to detect and sample spontaneous ruptured aortic plaques (SRAPs). We aimed to establish novel methods to detect cholesterol crystals (CCs) in sampled SRAPs. Methods Blood specimens containing SRAPs were obtained from patients using NOGA. Blood was instantly frozen on a glass slide and subsequently thawed for quantitative analysis and spread onto a filter paper that was rinsed using distilled water. Qualitative analysis was performed for the rinsed water using polarized light microscopy, and the filter paper was embedded in paraffin for histologic analysis. Results The CCs were clearly observed after hemolysis using the instant freeze-thaw method. The filter paper rinse method indicated free CCs of varying shapes under polarized light microscopy without erythrocytes. On the filter paper, sampled SRAPs showed Lamé-like small particles. Histopathology revealed various atheromatous components. Conclusion A set of novel methods for detecting CCs from sampled blood was established.

Sarcomatoid carcinoma of the urinary bladder is a rare entity, whose histogenesis and biological behavior remain controversial. The cytological literature on sarcomatoid carcinoma in voided urine is very scarce. Clinically, the diagnosis of this tumor can be made by computed tomography (CT), magnetic resonance imaging (MRI), cytology, and biopsy material. In this study, cytology, histopathology, and radiological imaging were employed in order to reach a diagnosis of sarcomatoid carcinoma. CT imaging showed increased thickness of the bladder wall associated to a polypoid mass. MRI showed a 4-cm sized, broadly necked polypoid mass with calcification and ulceration at the right side of the bladder wall. T2W1 imaging showed low signal. Voided urinary cytology showed a scattered cellular presentation. The tumor cells had a high nucleo- cytoplasmic ratio, with elongated cytoplasm with faint with indistinct cytoplasm border. The nucleus was oval to round, with large and irregular nucleoli and irregular nuclear membrane. These tumor cells were positive for cytokeratin (CKAE1/AE3), vimentin, p53, carcinoembryonic antigen (CEA), α1-smooth muscle actin (SMA) by the immunoperoxidase staining. Histopathology showed spindle-shaped and clumped large tumor cells with abundant cytoplasm. Mitotic figures were frequently seen and varied from area to area (50% of the tumor cells were positive for MIB1). Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.

The diagnosis of gastrointestinal stromal tumor (GIST) is generally established on histopathologic examination of surgical specimens. Gastrointestinal stromal tumor comprises a heterogenous group of neoplasms of the gastrointestinal tract previously referred to as leiomyomas, leiomyosarcomas, or schwannomas. Gastrointestinal stromal tumor arising from anorectum is a rare instance. We report a case of GIST for the correlation of imaging and cytologic features with immunocytochemical staining. A computed tomography and magnetic resonance imaging confirmed a 2-cm tumor growing into the rectal lumen. The central portion of the tumor showed T1-weighted imaging of low signal and suspected central necrosis by the T2-weighted imaging of high signal. Imprint cytology from excised tumors showed isolated or loosely aggregated spindle cells with scanty and fibrillary cytoplasmic processes, nuclear pleomorphism, fine granular chromatin, and irregular nuclear margins. Epithelioid tumor cells showed grooves with abundant cytoplasm and several round nucleoli. Both c-kit and CD34 antigen were positive with strong and diffuse stainability in smears as well as paraffin sections by immunoperoxidase staining. We suggest that the combined use of imaging diagnosis and cytology with immunocytochemical staining are useful initial diagnosis of GIST.

The mortality rate due to rupture of aortic dissection and aortic aneurysm is approximately 90%. Acute aortic rupture can be fatal prior to hospitalization and has proven difficult to diagnose correctly or predict. The in-hospital mortality rate of ruptured aortic aneurysm ranges from 53 to 66%. Emergency surgical and endovascular treatments are the only options for ruptured aortic dissection and aortic aneurysm. No method of systematic early detection or inspection of vessel injury is available at the prevention stage. Regardless of the improvement in many imaging modalities, aortic diameter has remained a major criterion for recommending surgery in diagnosed patients. Previous reports have suggested a relationship between vulnerable plaque and atherosclerotic aortic aneurysm. Non-obstructive angioscopy is a new method for evaluating intimal injury over the whole aorta. It has been used to identify many advanced atherosclerotic plaques that were missed on traditional imaging modalities before aneurysm formation. Non-obstructive angioscopy has shown that atherosclerosis of the aorta begins before that of the coronary artery, which had been noted on autopsy "in vivo". Strong or repetitive aortic injuries might cause sudden aortic disruption. Aortic atheroma is also a risk factor of stroke and perivascular embolism. Detecting aortic vulnerable atherosclerotic plaque on non-obstructive angioscopy may not only clarify the pathogenesis of acute aortic rupture and "aortogenic" thromboemboli and atheroemboli but also play a role in the pre-emptive medicine.

Feline immunodeficiency virus (FIV) infection in cats has been reported to be a useful animal model for human AIDS studies, especially in the early stages of infection. We examined the temporal changes in provirus detection in peripheral blood mononuclear cells (PBMC) and the distribution of FIV-DNA and RNA in feline tissues by the polymerase chain reaction at 10, 35, 70 days after intravenous inoculation of FIV. Viral DNA in the PBMC was detected three to four weeks after infection and its fluctuation was demonstrated for the first time. Ten days after infection, before seroconversion, proviruses were detected only in the mesenteric lymph nodes and intestines. At 35 and 70 days after infection, after seroconversion, proviruses were detected in most lymphoid organs and the salivary glands, but the expression of FIV-RNA was limited to the thymus at 70 days after infection. These results show that FIV-RNA is transcribed from proviral DNA exclusively in the thymus at this stage. We suggest that the quantitative changes in detectable proviruses in the PBMC depend on the relation between the decrease in infected cells caused by cytolytic T lymphocytes and/or apoptosis and their increase caused by the release of a new supply of lymphocytes from the thymus.

BAL細胞診にCMV感染細胞, カリニおよび腫瘍細胞が認められたATLの1例を経験したので報告する.BAL細胞診, 剖検肺組織標本を用いて, カリニ, CMV感染細胞の形態学的および組織細胞化学的検索を行った.カリニは, メイギムザ染色で嚢子内小体が, グロコット染色で嚢子壁, 嚢子内括弧状構造が染色され, PAS, グリドリーおよび酵素抗体法では嚢子壁が鮮明に染色された.また, 電顕では厚い嚢子壁, 栄養型およびtubular expansionが観察できた.CMV感染細胞は, 粘液染色で細胞質内封入体が顆粒状に陽性を示し, 酵素抗体法, ISH法では核縁, 核内封入体および細胞質内封入体が陽性を示した.さらに電顕にてVirus particleの存在を確認した.

Primary cardiac sarcomas are rare instances and only occasionally documented in the cytologic literature. Usually, the diagnosis of these rare lesions can be made at echocardiography, aspiration biopsy cytology, cardiac biopsy, and open cardiac surgery (intraoperative diagnosis). In this study, cytologic configurations and immunohistochemistry for 3 primary cardiac sarcomas (rhabdomyosarcoma, angiosarcoma, and malignant fibrous histiocytoma) were revealed. In rhabdomyosarcoma (right ventricle), the tumor cells exhibited an anisocytotic spindle-shaped nuclei with hyperchromasia and an obscure cytoplasmic margin. Vimentin and myosin were positive throughout the cytoplasm for the tumor cells. In angiosarcoma (right atrium), small clusters of anisocytotic spindle-shaped tumor cells appeared as vascular-like structures and hemosiderin-laden macrophages in many erythrocyte-rich backgrounds. Nuclei showed round to oval shape with hyperchromasia and prominent large nucleoli. Cytoplasm was obscure and elongated. Factor VIII related antigen and CD34 were strongly positive throughout the cytoplasm for the tumor cells. In malignant fibrous histiocytoma (right ventricle), the tumor cells exhibited oval to spindle-shaped and elongated nuclei and coarse granular chromatins with hyperchromasia. The nuclear margin was thin. A few small round nucleoli appeared. Elongated obscure and foamy cytoplasm was stained pale blue. Vimentin and α 1 -antitrypsin were positive throughout the cytoplasm for the tumor cells. This study elucidated the cellular characteristics and immunohistochemistry for cardiac sarcomas using imprint smears as an aid to cytopathologic diagnosis.

悪性リンパ腫患者の気管支擦過細胞診にてcytomegalovirus感染細胞の1例を経験したので報告する.症例は56歳の主婦で, 咳および皮膚の掻痒性発疹で入院した.皮膚生検を実施し, 病理組織学的にmycosis fungoidesの診断がなされ, ただちに, vincristin, endoxan, leukerin, predoninの化学療法を実施した.胸部X線で薬剤性, 癌性リンパ管炎あるいはカリニ肺炎を疑った.気管支擦過細胞診を実施し, cytomegalovirus感染細胞を検出し, 蛍光抗体法にて細胞質に特異蛍光を認めた.さらに, in situ hybridizationにてウイルスDNAを確認し, 細胞診断学上有用であった.

胎児水腫 (胎齢27週) の腹水細胞診を実施し, 核内封入体細胞を認め, ヒトパルボウイルスの感染を疑った1例を経験した.核内封入体細胞のN/C比は大から中等度であった.核は腫大し, 直径10～15μmであった.また, 核内封入体細胞は核縁が肥厚し, ハロー形成があり, 青紫色に染まるスリガラス状の核であった。細胞質は青紫色であった.酵素抗体法 (パルボウイルスB19に対するモノクロナール抗体) により感染細胞の細胞質に特異抗原を認めた.病理解剖時の脾, 肝, 腎, 心および胎盤の組織に核内封入体細胞を認めた.細胞診が伝染性紅斑の診断に有用と思われた.

The cytologic changes in the smears of fetus ascites fluid with parvovirus B19 infections are described. Cytology revealed the ground-glass appearance of the nuclei having a perinuclear halo, and the nuclear chromatin was spread as a thin rim around the inclusion. Subsequently, monoclonal antibodies for parvovirus B19 were applied to identify the specific antigen in the Papanicolaou-destained specimens. Positive staining reaction products were found throughout the cytoplasm of cells. Similar phenomena were confirmed in paraffin-embedded tissue sections of many organs in autopsy materials. It is hoped that recognition of those virus-infected cells in the body cavity fluid of the fetus will be helpful in making a diagnosis of hydrops fetalis associated with parvovirus B19 infections of the pregnant woman. © 1995 Wiley-Liss, Inc.

Papanicolaou-stained cervicovaginal smears from six patients with herpes simplex virus (HSV) infection were destained and reprocessed by means of in situ hybridization (ISH) technique to demonstrate the presence of HSV DNA utilizing biotinylated probe. Positive results were obtained in all six cases with intense staining signal for the HSV DNA in the nuclei of cells having a ground-glass nuclear appearance as well as in multinucleated giant cells. Furthermore, a hybridization signal was also noted in smears that had been prepared as much as 3 yr previously. HSV type 2-specific antigen was confirmed in six destained smears by means of immunoperoxidase staining. Moreover, polymerase chain reaction (PCR) was also performed for four patients from Pap-destained cervicovaginal smears. Amplified HSV DNA was detected in all four cases as 106 basepair PCR products by polyacrylamide gel electrophoresis. The combined use of cytology and the ISH technique and PCR amplification was of great value for the rapid cytodiagnosis of genital infection of HSV. Diagn. Cytopathol. 2003;29:246–249. © 2003 Wiley-Liss, Inc.

During the past 9 yr, 187,529 Sternheimer-Malbin-stained urinary sediments were examined as routine urinalysis specimens from patients attending the National Cardiovascular Center in Osaka, Japan. Abnormal cells were found in 20 patients who did not have clinical diagnoses of malignancy. Malignant cytological changes in 18 patients resulted in a rate of 1 case in 6,751 patients; the two remaining specimens with abnormal cells showed polyomavirus infection. This article describes our experience in the diagnosis of malignant cells of the urinary tract through the cooperation of the clinical and cytological laboratories. Since in Japan, the rate of death for bladder cancer is similar to 1 in 6,751, this method seems to be of great use in the diagnosis of urinary tract malignancies.

Using the immunoperoxidase technique, we studied "serosal balls," which have features resembling those of cells from primary and metastatic tumors, and may thus complicate cytodiagnosis. Serosal balls were detected in 32 (18%) of 174 peritoneal washings. The balls consisted of oval clusters of cells in solid masses surrounded by flattened cells. The interior of the serosal balls was stained green with Papanicolaou method, showing the presence of homogeneous amorphous material, sometimes stained in a filamentous pattern. Almost all serosal balls were stained immunocytochemically for both keratin and vimentin. The interior was stained with antibodies against collagen types I and III. Therefore, these balls were fragments of serous membrane, and contained fibrous tissue and mesothelial cells.

Cytological findings are presented of seven cases of cardiac myxomas. Avidin-biotin-peroxidase complex (ABC) method was employed to demonstrate Ulex europaeus agglutinin-I (UEA-I) lectin in imprint smears as well as in paraffin-embedded tissue sections in cardiac myxomas. The cytology was characterized by tumor cells with polyhedral or stellate and mucinous background with lymphocytes, neutrophilles, and hemosiderin-laden machrophages. In smears as well as tissue sections, UEA-I lectin was detected throughout the cytoplasm of myxoma cells. This study established the applicability of the immunoperoxidase staining for cardiac myxoma as an aid in cytopathological diagnosis.

Papanicolaou stained bronchial brush and imprint pulmonary smears containing intranuclear and cytoplasmic inclusion bearing alveolar pneumocytes suggestive of cytomegalovirus infection were destained and reprocessed for in situ hybridization using a biotinylated probe for cytomegalovirus DNA. Two cases were processed in this way. A hybridization signal for viral DNA was noted in each case. However, no reddish brown staining reaction products were noted in any of the control samples. This simple and rapid nonradioactive detection system is a valuable supplement to routine pulmonary cytology for the definitive diagnosis of this virus infection, and this technique is also appropriate for retrospective study. © 1992 Wiley-Liss, Inc.

Papanicolaou stained smears of urinary sediment containing inclusion bearing urothelial cells suggestive of human polyomavirus infection were destained and reprocessed for in situ hybridization using a biotinylated probe for human polyomavirus DNA. Seven slides were processed in this way. A hybridization signal for viral DNA was noted in each case, even in smears that had previously been stored for 11 years. This simple and rapid non-radioactive detection system is a valuable supplement to routine urinary cytology for the definitive diagnosis of this virus infection.

Typical herpesvirus keratitis that developed in a 12-year-old boy was initially diagnosed cytologically by the Papanicolaou method demonstrating the 'ground-glass' appearance of the nuclei with multinucleated syncytial cells. Subsequently, the in situ hybridization technique was applied to identify the herpes simplex virus (HSV) DNA in the Papanicolaoudestained cellular samples. Positive hybridization was found with intense staining for the HSV DNA in the nuclei of cells having a 'ground-glass' appearance. In situ hybridization has been shown to be a useful technique for the identification of HSV in corneal scrapes, and similar studies may be carried out in cellular samples from the other body sites.

褐色細胞腫5症例の捺印細胞像, 分泌顆粒検索の目的に酵素抗体法ならびに電子顕微鏡的観察を行った.1) 褐色細胞腫の特徴所見は上皮様の結合塊の配列がみられ, 連珠状の多核細胞および相互封入像が5症例中全例に認められ, 核内空胞は5症例中3例に認められた.2) 細胞質は広く, ライトグリーンで細顆粒状に染色された.3) 細胞, 核ともに大小不同が著明であったが, クロマチンの増量がみられるものの細顆粒状で, 均等分布を示した.4) 酵素抗体法では細胞質あるいは核内空胞にendocrine granule constituents (以下, EGC) が顆粒状に陽性を示した.5) 電子顕微鏡的観察で腫瘍細胞の細胞質に電子密度の高い分泌顆粒を確認した.以上のことから抗EGC抗体を細胞診に応用することは内分泌系腫瘍のスクリーニングに有用と思われた.

髄液細胞診で悪性細胞の存在を胃原発の3症例につき証明することができた.その細胞診所見と病理組織学的所見を比較検討した.症例1の髄液細胞像は孤立散在性で, 結合性も弱く, 重積性はない.細胞質は青緑色一部空胞形成がみられ, N/C比は大である.細胞径は10～20μ, 核径は12～13μで.比較的小型である.核小体は大きく丸い.症例1については病理解剖を実施し, 胃原発巣を大彎側幽門側に認め, 8×7×2cm大のBorrmann III型の腫瘍であった.組織学的には印環型腺癌であった.電子顕微鏡像は腫瘍細胞間の接合斑はみられず, 腺管形成がみられた内腔に粘液はなく, また, 微絨毛も明瞭でなかった.症例2の髄液細胞診は症例1とほぼ同様の所見を呈した.手術時に大彎側幽門側に直径5cmのBorrmann III型の腫瘍を認めた.組織学的には中等度分化型腺癌の浸潤部位と極めて分化度の低い腫瘍細胞の2つの細胞像が観察された.症例3では, 胃角部, 食道下端にBorrmann IV型の腫瘤の存在を認めた.病理組織診断は印環型腺癌であった.髄液細胞診は症例1および2と同様の細胞像を呈した.症例2および3についてはPAS染色を実施し, 原発巣推定に有用であった.

A 80-year-old female, who had a diagnosis of hepatoma with cirrhosis and showed gradually increasing level of α-Fetoprotein, was investigated cytologically, histopathologically and electron microscopically.Tumor imprint cytology disclosed adenocarcinoma-like appearance, that is, overlapped cells, cytoplasmic vacuoles and large, round nucleolei. Histopathological findings of tumor were consisted with glandular formation and PAS positive material in their lumens. And epitherial junctional complexes and glycogen granules revealed by electron microscope.We made a diagnosis of this case as a mucin-producing hepatoma and discussed differential diagnosis between adenocarcinoma such as cholangiocellular carcinoma or hepatocellular carcinoma especially from the point of cytological details.

Myoclonus is a characteristic neurological sign of subacute sclerosing panencephalitis (SSPE). Attempts were made to induce myoclonus in a large proportion of hamsters with a cell-associated strain of SSPE virus (the Biken strain) and thereby to establish an experimental model for study of the mechanism of development of this condition. When injected intracerebrally, Biken virus induced myoclonus within two to 14 days in 8407o of the three- to nine-week-old hamsters tested. Electroencephalographic traces showed a periodic and synchronous discharge consisting of high-voltage slow waves and spikes that appeared coincidentally with myoclonus. Neurons in the cortex and thalamus of the affected animals had severely degenerated cytoplasm. Inflammatory changes, such as perivascular cuffing or infiltration of mononuclear cells, were not detected. Staining with immunoperoxidase revealed measles viral antigens in the cytoplasm and dendrites of the affected neurons. SSPE virus with the same properties as the parent virus was recovered from brain cells of sick animals by cocultivation with Vero cells.

Papanicolaou-destained imprint smears from 24 brain tumors were investigated by means of avidin-biotin-peroxidase complex method (ABC) with the use of monoclonal antibodies against glial fibrillary acidic protein (GFAP). Positive staining reaction to GFAP antibody has been demonstrated in cells from the following tumors: astrocytoma, anaplastic astrocytoma, glioblastoma multiforme, mixed glioma, and ependymoma. The reaction for GFAP was negative for the following tumors: medulloblastoma, neurilemmoma, melanoma, hemangioblastoma, and metastatic tumors. In astrocytoma, the cell bodies and processes were positive with delicate fibrillary patterns; in anaplastic astrocytoma, cytoplasm and the processes were intensively stained. In glioblastoma multiforme, the staining patterns were also mixed, and the short, thickened processes were characteristic. Use of both a smear preparation and the immunoperoxidase staining technique is of great value in diagnosis of tumors of the central nervous system.

1970年, 健康なシチメンチョウ105羽中6羽の血液から, アヒル胎児線維芽細胞を用いて, ヘルペス型ウイルスを分離した. これらのウイルスでは, マレック病ウイルスと異なり, 細胞遊離ウイルスを容易に得ることができた. 特にO1株において, その傾向は明瞭であった. 感染培養細胞には, プラークが認められ, プラークは, 多数の多核巨細胞より成っていた. 感染細胞のヘマトキシリン・エオシン染色において, ヘルペス型の核内封入体の形成が認められた. この封入体は, Feulgen 反応およびアクリジン・オレンジ染色により, DNA 陽性を示し, 3H-thymidine のオートラジオグラフィーにおいて, その中にヘルペス型配列の銀粒子が認められた. また DNA 合成阻害剤に対し感受性を示した. さらに感染細胞の超薄切片を電子顕微鏡で観察すると, ヘルペス型ウイルスの構造をもつ粒子が, 核内に多数認められた. これら二重膜粒子の直径は約140mμ, カプシドの直径は, 約100mμであった. しかし, マレック病ウイルス Biken C2 株感染細胞の場合と異なり, カプシドの内部が十字紋様を呈するものが多数認められた. また, 感染材料を燐タングステン酸によりネガティブ染色して, 電子顕微鏡で観察すると, 中空のカプソメア(推定162個)より成るヘルペス型カプシド構造が認められた. これらのウイルスは, 分離当初よリアヒルおよびウズラ胎児線維芽細胞上におけるプラーク形成が明瞭であり, 培養には多核巨細胞も出現した. アヒル, ウズラおよびニワトリ胎児線維芽細胞は, このウイルスに対して, ほぼ同一の感受性を示した. 螢光抗体法により, マレック病ウイルスと交差反応を呈したが, 同一反応ではなかった. 以上の研究により, 本ウイルスは, マレック病ウイルスと異なり, Witter らの報告したシチメンチョウ・ヘルペスウイルスと性格がほぼ一致するものと考えられる. 本研究の要旨は, 第71回(1971年4月)および第72回(1971年10月)日本獣医学会において報告した. 終わりに臨み, 電子顕微鏡による研究について, 御指導御協力下さった当研究室新居志郎助教授, 大阪医科大学内藤継也氏, ならびに本研究に際し, 種々御協力を賜わった兵庫県経済連藤田重夫部長, 小馬栄猛氏, 岸田満須夫氏, および柏原農協永井明男氏に深謝する.

A study was undertaken to evaluate the cellular findings of gastric carcinoma in cerebrospinal fluid (CSF). Immunocytochemical localization of carcinoembryonic antigen (CEA) was performed on four cases of metastatic gastric carcinoma cells in CSF samples. A positive peroxidase-antiperoxidase (PAP) reaction was obtained in all cases, with intense staining for CEA in the CSF samples as well as in the paraffin-embedded tissue sections from the primary gastric tumors. Cellular morphology and the results of immunoperoxidase staining can be studied simultaneously. We believe that the PAP method for CEA increases diagnostic accuracy of cytology in the CSF.

A case is presented in which malignant squamous cells and herpes virus infected cells were recognized concomitantly at routine cytologic examination for detection of cervical cancer. Further examinations on admission revealed Stage 1b carcinoma of the cervix and characteristic pathologic changes of herpetic infection in the tumor cell nests. Virus was isolated from the cancerous tissues in the ectocervix following radical hysterectomy, and was identified as herpes simplex virus type 2 (HSV-2). The typical HSV-2 particles were detected by electron microscopic observation on the infected FL monolayer cells with isolation. Neutralizing antibody levels against HSV-2 declined and remained low after hysterectomy. The HSV-2, isolated and identified, might be considered as "passenger" virus. The role of this isolated virus as an etiologic agent of the cervical cancer was not clear.