Fluvastatin suppresses atherosclerotic progression, mediated through its inhibitory effect on endothelial dysfunction, lipid peroxidation, and macrophage deposition.

Fluvastatin, a potent 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, exerts an inhibitory effect on intimal thickening after mechanical injury in normocholesterolemic rabbit artery at a dose not enough to elicit a known action of lipid lowering. This study was designed to determine whether atherosclerotic progression triggered by hypercholesterolemia can be inhibited by fluvastatin under conditions without its hypocholesterolemic effect. Rabbits were fed a 0.5% cholesterol diet or normal diet for 17 weeks and were treated with either fluvastatin (0.3-2 mg/kg/day, p.o.) or pravastatin (2 mg/kg/day, p.o.). Atherogenic features manifested in the cholesterol-diet group, compared with the normal-diet group; they were the increase in serum lipid peroxide level, in the intraluminal lesion area of the aorta, and in macrophage content of the aortic cross-sectional lesion area; the attenuation of endothelium-dependent relaxing response to acetylcholine in the femoral artery; and the increase in serum lipid level. Treatment with fluvastatin, but not pravastatin, inhibited the manifestation of the atherogenic features without a serum lipid-lowering effect. Thus fluvastatin is likely to reduce the risk of atherosclerotic progression, to which endothelial dysfunction, lipid peroxidation, and macrophage accumulation in the vasculature may contribute, irrespective of changes in serum lipid levels.

[Immunohistochemical study of Sertoli-stromal cell tumor; comparison between the tumor arising from the gonad of a testicular feminization syndrome bearing patient and from ovaries of non-bearing patients].

The association of Sertoli-stromal cell tumor with testicular feminization syndrome (TFS) has been elaborated in the past studies. Here, we described immunohistochemical studies on Sertoli cell tumor of the gonad in a TFS patient and compare with 2 other cases of spontaneous ovarian Sertoli-stromal tumor. [Case 1] The case was a 73 year-old Japanese patient (46XY karyotype), who had had primary amenorrhea. High level of testosterone was noted in laboratory investigation (1900 ng/ml). No ambiguous morphology of external genitalia was present, but atrophy of vagina was noted. The patient was diagnosed as TFS. A left gonadal tumor was identified histologically showing well differentiated Sertoli cell tumor. The tumor cells were positive for anti-vimentin antibody but negative for anti-keratin, EMA and p53 antibodies by immunohistochemistry. The right gonad was an immature testis. [Case 2] The case was a 33 year-old female with ovarian Sertoli-Leydig cell tumor. Immunohistochemically, positive reaction for anti-keratin and p53 antibodies were observed. [Case 3] The case was a 17 year-old female with moderately differentiated Sertoli cell tumor of the ovary. The tumor cells were positive for anti-keratin, EMA and p53 antibodies by immunohistochemistry. Difference in immunohistochemical reactions between Sertoli cell tumor in TFS and Sertoli-stromal cell tumors of the ovaries was probably due to variation in the degree of gonadal development.

Expression of apoptosis and its related protein in astrocytic tumors.

The relationship between malignant potential and apoptosis in astrocytic tumors has not been clearly defined, and further classification of astrocytic tumors is necessary. To elucidate the relationship between the histopathological grade of astrocytic tumors and apoptosis, we studied 25 cases of astrocytic tumors, comprising 10 cases of glioblastoma (GB), 7 cases of anaplastic astrocytoma (AA), and 8 cases of astrocytoma (AC). We detected apoptosis using the TdT-mediated dUTP-biotin nick-end labeling (TUNEL) method. We studied immunohistochemical expression of bcl-2 protein and p53 protein, which are apoptosis-related factors, and cell proliferative activity using Ki-67 antibody. No significant change was noted between apoptotic index and the histological grade of the tumors. In GB, apoptotic cell-rich foci were present at the pseudopalisading necrosis. No correlation between histopathological grades and expression of either p53 or bcl-2 was observed. In GB, however, poor distribution of bcl-2 was found in the areas of pseudopalisade formation. bcl-2 is one of the regulatory factors in the cell cycle and inhibits apoptosis. Expression of apoptosis had no correlation with histopathological grade. However, in GB, the distribution of apoptotic cells showed a correlation with bcl-2-poor foci. It was thought that apoptosis was one of the regulatory factors in the formation of pseudopalisading necrosis in GB.

[Usefulness of cytology applied simultaneously to frozen section at rapid intraoperative diagnosis of intracranial tumors].

We discussed usefulness of cytology for rapid intraoperative diagnosis of intracranial tumors using frozen section. Twenty five cases of intracranial tumors (6 cases of astrocytoma, 6 cases of glioblastoma multiforme, 4 cases of meningioma, 2 cases of pituitary adenoma, 1 case of ependymoma and 6 cases of metastatic carcinoma) were observed by cytological specimen, frozen section and formalin fixed-paraffin embedded section. When only frozen section were used for histopathological diagnosis, it was difficult to diagnose in a few case of astrocytic tumors by artifact in freezing process. When only cytological specimen were used for diagnosis, histological findings, for example pseudopalisading around necrosis in glioblastoma or rosette formation in ependymoma, were not identified. However, diagnostic accuracy were improved when both cytological and frozen specimen were used if adequate material were obtained for diagnosis. In conclusion, cytological approach used in addition to frozen section was useful for rapid intraoperative diagnosis.

[Adult multiple coronary aneurysms of Kawasaki's disease's sequelae; two autopsy cases].

Coronary aneurysm in Kawasaki's disease (Acute febril infantile mucocutaneous lymph node syndrome, MCLS) may cause sudden death in childhood and ischemic heart disease in adults. We encountered two adult autopsy cases of Kawasaki's disease with multiple coronary aneurysms. The first case was a 56-year-old man who hospitalized due to recurrent syncope since 51 years of age. At age 55 coronary angiography (CAG) had shown multiple aneurysms in the left and right coronary artery. In September 1991, he developed chest pain and was brought to the hospital, almost dead on arrival (DOA). The patient died later the same day despite cardiopulmonary resuscitation. Autopsy findings showed cardiomegaly (470 g) with multiple coronary aneurysms of three coronary arteries. Microscopically, intimal thickening and medial thinning were found in the aneurysmal wall with calcification and disruption of the internal elastic lamina. The second case, a 28-year-old man had been diagnosed with rheumatic fever and mitral regurgitation at 4 years of age. Coronary aneurysms were noted on CAG at 26 years of age. In April 1992, he developed fever and was admitted to a local hospital where he was diagnosed with infectious endocarditis. After his being transferred to our hospital, disturbance of consciousness suddenly developed and he died in September 1992. Autopsy findings showed cardiomegaly (430 g) with left ventricular hypertrophy and multiple coronary aneurysms in left anterior descending coronary artery and left circumflex coronary artery. The aneurysmal wall showed intimal thickening and medial thinning with multiple recanalizations of occlusive lumina and fibrous intimal thickening. The mitral valve showed mild fibrosis and calcification without valvular deformity. There was no evidence of bacterial endocarditis. Both cases were finally diagnosed as Kawasaki's disease. Ischemic heart disease or lesions related to coronary aneurysm in Kawasaki's disease may show an increased incidence in the near future. Kawasaki's disease should have been followed even in adulthood after treatment in childhood.

[Method of quantitative measurement of human platelet aggregation in clinical practice].

We examined the quantitative measurement of platelet aggregation in 20 volunteers and 89 patients with ischemic heart disease (IHD). Subjects in whom platelet aggregation was induced by an adenosine diphosphate (ADP and an epinephrine solution were clearly divided into two groups: "normal" and "accentuated". We chose the maximum aggregation time for the ADP solution and the maximum transmission rate for the epinephrine solution for the quantitative measurement of platelet aggregation. The results were as follows. The maximum aggregation time with a 1.0 microM ADP solution in the normal group of 13 volunteers was 0.71 +/- 0.12 min, (mean +/- SD), in 50 IHD patients it was 0.91 +/- 0.49 min, in the "accentuated" group of 7 volunteers it was 5.34 +/- 1.18 min, in 39 IHD patients it was 6.01 +/- 1.22 min. The maximum light transmission rate with a 0.1 microM epinephrine solution in the normal group of 14 volunteers was 8.04 +/- 4.14%, in 52 IHD patients it was 13.74 +/- 5.75%, in the "accentuated" group of 6 volunteers it was 76.33 +/- 9.91%, and in 37 IHD patients it was 77.26 +/- 13.39%. The difference between the "normal" and "accentuated" groups was statistically significant (p < 0.001), with the ADP and with the epinephrine solution. Next, we examine the effect of antiplatelet therapy (dipyridamole and aspirin) on 15 patients selected from among those with IHD who had accentuated platelet aggregation when the ADP solution was added. Their maximum aggregation time was 5.98 +/- 1.24 min, and their maximum platelet aggregation rate when the epinephrine solution was added was 78.90 +/- 11.60% (10 patients), 20.00 +/- 3.24% (5 patients). After the therapies, the maximum aggregation time in 10 patients was 0.94 +/- 0.21 min (return to normal condition), and in 5 patients it was 5.90 +/- 1.09 min (not effective). The maximum platelet aggregation rate in 10 patients was 14.11 +/- 5.88% (normal condition), and in 5 patients it was 75.00 +/- 9.51% (not effective). Finally, we examined the long term effects of various antiplatelet drugs. Most of the patients in whom platelet aggregation was accentuated got well.

Correlation of two argyrophilic nucleolar organizer region counting methods with the Ki-67 labeling index in uterine smooth muscle cell tumors.

The argyrophilic nucleolar organizer regions (AgNOR) are silver stained granules that are thought to correlate with cell proliferation activity. Two AgNOR counting methods: the mean AgNOR count (mAgNOR, the mean number of AgNOR granules in 100 cells) and the AgNOR proliferative index (pAgNOR, the percentage of cells exhibiting five or more AgNOR granules per nuclei) have been proposed. In this study, the two counting methods were applied to 58 cases of normal uterine corpus and uterine corpus tumors and were compared with the Ki-67 labeling index (Ki-67 LI) using MIB-1 monoclonal antibody and other histopathological criteria. Notable differences in the number of AgNOR and the Ki-67 LI were observed between benign and malignant smooth muscle tissue. Histopathologic features are well correlated to the proliferative activity of tumors. Although the most reliable method of predicting malignant potential cannot be determined, the methods outlined by this study are thought to be highly useful in assessing proliferative activities.

[Quality assurance of pathology laboratories: analysis of histopathological diagnosis of endoscopical biopsies from lower alimentary tract based on specimen types].

As a part of the internal quality assurance in the pathology laboratories, we focused on the diagnosis of biopsies from the lower alimentary tract, and investigated the number of independent lesions contributing the specimens, biopsy pieces, and paraffin blocks prepared in each pathological examination, from the standpoint of the types of biopsy procedures employed. In addition, we compared those of pre- and post-operative diagnoses, and examined the adequacy in the cases with repeated biopsies. Regarding the procedures, polypectomy and EMR (endoscopical mucosal resection) accounted for more than 60% of all the specimens handled during the study period. Most of the specimens taken by polypectomy or EMR were derived from neoplastic conditions, and roughly 40% of them were from malignancies including borderline lesions in a narrow sense. The examination of the surgically resected specimens confirmed a hundred percent accuracy ratio of the diagnosis on the biopsy specimens. Three cases (7.1%) out of 42 malignancies resected needed repeated biopsies in preoperative confirmation of malignancy, while the remainder (92.9%) had been diagnosed at the initial biopsy. The result disclosed that biopsies possessing both diagnostic and therapeutic value (polypectomy and EMR) are apparently increased in the field of lower alimentary tract, and the difficulty and efforts required for diagnosis are quite different and depend on the biopsy procedures employed.

[Immunohistochemical study of proliferation index of astrocytomas classified by new histopathological grading].

A new grading method of astrocytoma, proposed by Daumas-Duport et al, was based on four criteria:nuclear atypia, mitosis, necrosis and endothelial proliferation (0 criteria = Grade 1, 1 criteria = Grade 2, 2 criteria = Grade 3, 3 or 4 criteria = Grade 4). To elucidate the correlation of the classification based on four criteria, cellularity and proliferative activities, we studied proliferating index of 67 astrocytomas, using immunohistochemical staining for Ki-67 antibodies (MIB-1). A new grading method was related to proliferating index in this study, however, cellularity of tumor cells was not related to proliferating index. In this study, endothelial proliferation was invariably seen in high grade(Grade 3 and 4) astrocytomas. These tumor vessels were associated with dysfunction of blood-brain barrier and some growth factors accelerated to tumor cell proliferation. Relation between histopathological grade of astrocytoma and proliferation index was supposed, and new grading system was thought to be useful for routine pathological examination.

[An evaluation study on arbekacin for MRSA-infectious diseases including pneumonia, septicemia and others].

Availability of arbekacin (ABK) was analyzed in the chemotherapy of 24 MRSA-infected patients with symptoms of pneumonia (12), sepsis (6) and others (6). Most patients had background diseases such as malignant tumors or cerebrovascular disorders. 47% (7/15) of them were immunologically abnormal. 17 of them had been previously treated with cephems, imipenem, minocycline or fosfomycin. The ABK therapy was performed with doses ranging 50-400 mg a day, divided into 1-3 times (mostly 100 mg x 2), and for 5-24 days. (18 patients were treated between 5 and 14 days). 14 patients (58%) received combined therapy with other antibiotics (mostly with beta-lactams, 12). The clinical efficacy rate of the ABK therapy was 62% (good, 13; fair, 4; ineffective, 4; unknown, 3). The bacteriological efficacies were: eradicated, 7 (44%); decreased, 4; no change, 5; unknown, 8. Side effects were found in 3 patients (oliguria, 2; eruption due to drug, 1) and one case resulted in serious renal disorder. Abnormal laboratory data were found in 7 cases. Above results have indicated that ABK is a useful antibiotic in chemotherapy of MRSA-infections.

[Elective course "Anatomic Pathology"].

The elective course "Anatomic Pathology" consists of three domains including (1) autopsy, (2) surgical pathology (biopsy) and (3) cytology. The major objectives are to gain knowledge and master various technics to make a high quality pathomorphological diagnosis. The training duration is fundamentally 2 years, but can be shortened to 1 year. The GIOs and SBOs of each domain are listed separately followed by specific learning subjects such as special stains, immunohistochemistry, electron microscopy, microbiology and forensic pathology. The major diseases in the various systems and organs are included. The minimal requirement for each domain is also mentioned.

[Immunohistopathological analysis of estrogen receptors in human meningioma].

We studied immunohistochemically estrogen receptors (ER) in 25 meningiomas removed from 20 patients using formalin-paraffin sections. Estrogen and progesterone receptors were studies in three case of meningiomas using frozen sections, also. Only two cases show positive reactions for estrogen receptor using formalin-paraffin sections. No correlation was found between estrogen receptor state and histological type, or biological behavior except for sex. Frozen sections failed to show the positive reaction for estrogen or progesterone receptors at all. The data suggest that estrogen receptors are detected in a few of the case with meningioma. Immunohistochemical analysis for estrogen receptor do not offer the useful informations in histopathological diagnosis or estimation of biological behaviors for meningioma on routine laboratory works.

[Roles of pathology laboratories on medical care service and quality assessment system for pathological diagnosis].

In practical medicine, pathologists provide highly specialized medical information (pathological diagnosis) to clinicians. Pathological diagnosis includes histopathology, cytology, intraoperative consultation (rapid diagnosis by frozen section) and autopsy service. In particular, histopathological diagnosis of biopsy specimens, nearly always gives a qualitative diagnosis to unknown pathological process, in other words, a final diagnosis. In addition, pathological diagnosis contributes to supporting medical audit. Thus it is quite important that to establish the quality assessment system for pathological diagnosis which is hard to be judged with the value.

[A study of intraoperative rapid frozen section diagnosis focusing on accuracy and quality assessment].

Accuracy of frozen section (FS) diagnoses was investigated in a consecutive series of 1129 intraoperative consultations. In our series, the correct diagnosis including clinically not serious minor errors was made in 96.9% of the cases. Whereas the accuracy ratio of FS diagnosis for unknown pathologic process decreased to 92.2%. Among all the types of tissues, central nervous system was shown to be the most frequently handled for FS, followed by lung/bronchus, breast, liver/biliary tract, lymph node and so forth. The requesting ratio of FS, on the other hand, tended to be high in the following specialties; neurosurgery (46.5%), thoracic surgery (19.0%), general surgery, (10.0%). Deferred diagnosis with a provisional diagnosis and misinterpretations in histology typing without serious clinical problem accounted for 2.7% and 3.1% respectively. Causes of erroneous diagnoses seemed to be multifactorial, such as inappropriate sampling, diagnosis on poor quality histology sections, lack of clinical information, lack of enough experience in FS practice of pathologists, or a combination of more than two of them, though inevitable cases showing minimal cytological and structural atypia were included. Intraoperative consultation by FS diagnosis is now essential to serve a good quality medication to patients. It is thus necessary that to establish a tight peer review system and also to provide an education program with regard to practice in FS diagnosis particularly for young pathologists.

[Argyrophilic nucleolar organizer region proteins (Ag-NORs) in human central nervous tumors].

A silver colloid staining technique for identifying nucleolar organizer region-associated proteins (Ag-NORs) was applied to 17 primary gliomas and 16 meningiomas. These comprised 8 glioblastomas, one pleomorphic xanthoastrocytoma, 8 benign astrocytoma, 10 nonrecurrent meningiomas and 6 recurrent meningiomas, in which the mean number of Ag-NORs' per nuclei, were 3.37, 2.34, 2.57, 2.16 and 2.80 respectively, identifying significant differences among these groups. Ag-NORs of four giant cell glioblastomas showed low numbers of Ag-NORs, suggesting better prognosis. In spite of the time consuming and complicated counting method Ag-NORs is reproductive and useful as a tool for estimating the proliferating potential of various brain tumors, and appears to widely applicable in clinical laboratories.

[Current alteration of Helicobacter pylori in gastro-duodenal diseases].

The current alteration of the detection rate of Helicobacter pylori (H. pylori) on mucous membranes of the stomach has been surveyed and reviewed for 49 cases of gastro-duodenal diseases. The survey has been conducted for 3 years and an endoscopic examinations have been performed 2-8 times. On the first visit, the diagnosis can be made endoscopically, and H2 blockers and another agents were used as therapeutic drugs. The detection rate was 67.3% (33 cases) for positive cases and positive reaction cases, while the rate was 32.6% (16 cases) in negative cases and negative reaction cases. Among them, the most commonly observed disease was atrophic gastritis, followed by duodenal ulcer, and acute gastric mucous lesions (AGML). Peptic ulcer was not seen. After administration of H2 blockers and anti-ulcer agents, the H. pylori-detection rate was found to fluctuate according to the diseases and alterations in the morbidity period. Some cases indicated the disappearance of H. pylori in accordance with the improvement of the lesion site.

Three-dimensional reorganization of a cell line of papilla Vateri adenocarcinoma in various culture conditions.

SAV-I, a cell line derived from a well differentiated adenocarcinoma of Vater's papilla, was cultured under four different conditions using collagen gel matrices (type I collagen): 1) double-layered, 2) floating double-layered, 3) embedded, and 4) floating embedded, then observed by light and electron microscopy and immunohistochemistry. Under all four conditions, three-dimensional growth with tubules occurred. In particular, the floating double-layered condition, where the cells were cultured between two collagen gel layers, then floated onto the medium, was useful for showing cellular reorganization. The three-dimensional growth patterns observed in vitro closely resembled the in vivo growth of SAV-I cells transplanted into nude mice. Therefore, we conclude that the floating double-layered condition is useful for demonstrating the morphological characteristics of the parent cells of established cell lines, and should be advantageous for studies of the relationship between cellular morphology and function in vitro.

Primary osteosarcoma of the ovary. A case report.

A case of primary osteosarcoma arising in the left ovary of a 75-year-old female is described. The chief complaint was a sensation of lower abdominal mass. An abdominal plain film showed a large calcified mass in pelvic region, and a preoperative diagnosis of "ovarian fibroma" was made. The excised tumor was divided into 4 pieces, resembling an oyster shell. Microscopically, the tumor fragments were composed of compact bone or woven bone with surrounding atypical osteoblasts and osteoclasts. The tumor was partly composed of numerous spindle cells with malignant osteoid or atypical chondroid formation, and diagnosed as "osteosarcoma". The cystic part of the lesion was lined with a single layer of columnar cells, but the tumor contained no other germ elements or stem cells, or malignant epithelium. Therefore, it is doubtful that this tumor originated from teratoma or malignant mixed mesodermal tumor, and we conclude that this ovarian osteosarcoma arose through a neoplastic change in ovarian stromal cells. The patient died 4 months after surgery due to intra-abdominal and intrathoracic dissemination of the tumor.

[Application of crush cytology preparation for rapid diagnosis of brain tumors].

The crush cytology perform during the surgery for 22 cases with brain tumors at Surugadai Nihon University Hospital, being compared with frozen and permanent histology sections. Crush cytology, which was generally and less damaged by artefacts than frozen sections. Immunocytochemistry used crush cytology were useful tools for differential diagnosis between undifferentiated glioma and carcinoma. Crush cytology were thought to be useful using together with the frozen sections for intraoperative diagnosis.