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背景 : 後腹膜に発生する後腹膜粘液性嚢胞性腫瘍 (Retroperitoneal mucinous cystic tumor) はまれな腫瘍である. 今回, 術前の穿刺細胞診で診断が困難であった 1 例を経験したので, 術中に得られた嚢胞内溶液の細胞像を示し, 考察を加え報告する.症例 : 20 代女性. 右下腹部痛にて当院に来院し, 腹部超音波検査で右側腹部嚢胞性腫瘤が認められた. 経皮的穿刺吸引液を用いて細胞診が 3 回行われたが, 粘液様物質が吸引されたものの, 上皮性細胞は認められず, 確定診断が困難であった. 開腹手術が行われ, 術中採取された茶褐色粘液状検体を用いて細胞診標本が作製された. 細胞所見では, 粘液性背景を伴って腺細胞の集団が多数認められた. 腺細胞は高円柱状で細胞質には粘液を認め, シート状, 索状, 乳頭状に配列していた. 組織所見では, 嚢胞内面に腺上皮細胞が, 平坦にあるいは乳頭状に増生していた. 間質への浸潤はなく, 低悪性度粘液性嚢胞性腫瘍と診断された. 腫瘍の間質細胞は, 免疫染色で「卵巣間質細胞」と同じ染色性を示した.結論 : 後腹膜の嚢胞性腫瘤の穿刺細胞診で, 粘液が認められた場合は, 上皮細胞が認められなくても鑑別診断すべき疾患のなかに粘液性嚢胞性腫瘍を入れておく必要がある.

We have developed a new test for estimating the secretory capacity of parathyroid hormone (PTH) from the parathyroid gland. Sodium bicarbonate solution [8.4% (w/v); 35 ml/m(2) body surface area] was infused for 2 min, and blood samples for the determination of plasma ionized calcium, plasma PTH (intact, midregion, carboxy-terminus) and related parameters were serially obtained. In 8 healthy volunteers, the mean (+/-SE) plasma ionized calcium fell promptly and significantly (from 1.21 +/- 0.01 to 1.11 +/- 0.01 mmol/L) after the sodium bicarbonate infusion. The mean (+/-SE) plasma intact PTH increased promptly and significantly, by more than four fold (42.3 +/- 4.2 to 182.4 +/- 34.7 pg/ml), and then gradually returned to basal levels. In patients with partial hypoparathyroidism who have detectable basal plasma levels of PTH, the absolute increment in PTH levels was much less, and in the plasma obtained from patients with complete hypoparathyroidism, absolutely no response was observed. Plasma obtained from patients diagnosed with primary hyperparathyroidism (parathyroid adenoma or hyperplasia) has high basal PTH levels. The response to the sodium bicarbonate infusion in these patients was markedly blunted (less than a two-fold increase in all cases examined). No significant adverse effects were observed during the procedure. Therefore, the sodium bicarbonate infusion test is a simple and sensitive method to stimulate PTH release, and is clinically useful for evaluating parathyroid gland function.

Background: Preoperative biopsy specimens obtained by means of percutaneous transhepatic cholangioscopy are useful for planning curative resection of bile duct carcinoma in an effort to improve survival. However, tissue diagnosis is sometimes difficult. This study evaluated the usefulness of p53 immunostaining of cholangioscopic specimens for determination of tumor spread. Methods: A total of 107 biopsy specimens from 28 patients with bile duct carcinoma was selected. Before surgery, these specimens were diagnosed histopathologically (hematoxylin and eosin staining) as positive or negative for carcinoma. After definitive surgery, specimens were immunostained with anti-p53 antibody. Results: Eighteen of 28 cases (64%) were positive for p53. Among these, 86% obtained from the main carcinomatous lesion or an area of superficial spread of the carcinoma exhibited a p53 labeling index (LI) over 25% as opposed to a p53 LI under 25% for all specimens obtained from noncarcinomatous lesions. Twelve specimens from 8 cases were classified before surgery as indeterminate (hematoxylin and eosin staining). The criterion of p53 LI over 25% was applicable in 11 of the 12 specimens. Conclusion: The p53 immunostaining of biopsy specimens obtained by means of percutaneous transhepatic cholangioscopy is helpful in determining tumor spread in bile duct carcinoma. (Gastrointest Endosc 2002;56:688-95.)

Of 987 cases of gastric adenocarcinoma seen at Nagoya University School of Medicine, we found 6 rare, extremely well-differentiated advanced gastric adenocarcinomas that could not be diagnosed as malignant tumors with only H&E staining, even with repeated biopsies under preoperative endoscopy. The aim of this study was to determine whether an immunohistochemical method using p53 and Ki-67 antibody would be helpful for preoperative pathologic diagnosis. The cancer control cases were 16 cases of ordinary well-differentiated advanced gastric adenocarcinoma, while the gastritis control cases were 22 cases of Helicobacter pylori–positive chronic gastritis. The p53 labeling index and the localization of Ki-67+ cells showed that the special adenocarcinomas in biopsy specimens were distinct from the surrounding normal mucosa and chronic gastritis, but not from the cancer control cases. These methods are useful markers for preoperative pathologic diagnosis of extremely welldifferentiated gastric adenocarcinoma, which sometimes is confused with regenerative atypical glands before operation. Diagnosis of gastric adenocarcinoma has not been difficult owing to technical advances in radiography, endoscopy

The possible role of calcineurin in the attenuation of cardiac hypertrophy and fibrosis by blockade of the angiotensin II type 1 (AT 1 ) receptor was investigated in Dahl salt-sensitive (DS) rats. The effect of the calcineurin inhibitor FK506 was also studied. DS rats progressively developed severe hypertension when fed a diet containing 8% NaCl from 7 weeks of age. In addition, marked cardiac hypertrophy and fibrosis were apparent and the activity of calcineurin and its mRNA expression in the myocardium was increased in these animals at 12 weeks in comparison with age-matched Dahl salt-resistant rats. The abundance of angiotensin-converting enzyme (ACE) and transforming growth factor (TGF)-β1 mRNAs was also increased in the hearts of DS rats at 12 weeks. Treatment of DS rats with a non-antihypertensive dose of the selective AT 1 receptor blocker candesartan (1 mg/kg per day) or FK506 (0.1 mg/kg per day) from 7 to 12 weeks attenuated both calcineurin activity and its mRNA expression in the heart, as well as the development of cardiac hypertrophy and fibrosis, without affecting cardiac function. Treatment with candesartan, but not FK506, prevented the upregulation of ACE and TGF-β1 gene expression. Both candesartan and FK506 prevented the load-induced induction of fetal-type cardiac genes. These results demonstrate that AT 1 receptor blockade attenuates the development of cardiac hypertrophy and fibrosis as well as the activation of calcineurin, without an antihypertensive effect, in rats with salt-sensitive hypertension. Calcineurin may be downstream from TGF-β1 in AT 1 receptor-mediated angiotensin II signaling in vivo.

To investigate the possibility of in vivo development of decreased vancomycin susceptibility, the vancomycin susceptibilities of 12 methicillin-resistant Staphylococcus aureus (MRSA) isolates serially recovered from six patients with vancomycin therapy were tested by standard MIC determination method and population analysis. While all of the MRSA isolates were susceptible to vancomycin (MICs, 1-2 microg/ml) by standard method, population analysis showed the upward shifts indicating decreased vancomycin susceptibility among serial isolates from two patients. These bacteria with decreased vancomycin susceptibility could be selected by using vancomycin selection of pre-therapy isolates under laboratory conditions. Furthermore, the reversion phenomenon of decreased vancomycin susceptibility was confirmed after 20 serial passages of the post-therapy isolates on drug-free agar. These data suggest that in vivo isolates may develop decreased vancomycin susceptibility that is not of such magnitude to cross a breakpoint threshold. This resistance may be unstable, and appears to result from a selective or inducible process that occurs in MRSA clinical strains during vancomycin therapy.

In this report, we describe a highly unusual case of glandular malignant peripheral nerve sheath tumor presenting as a neck mass in a previously healthy 29-year-old man. Grossly, the tumor was found to arise from a swollen peripheral nerve trunk. The tumor was largely composed of spindle cells that demonstrated marked nuclear pleomorphism and numerous abnormal mitotic figures. In addition, histologically malignant glandular structures lined by simple nonciliated columnar cells with goblet cells were found clustered in the center of the tumor. Examination of the swollen peripheral nerve trunk revealed the presence of a plexiform neurofibroma. The spindle cells were positive for S100. The glands were negative for S100 but positive for keratin, epithelial membrane antigen, and neuroendocrine markers (somatostatin, chromogranin, Leu-7, and calcitonin). This patient was subsequently diagnosed as having von Recklinghausen disease and died of tumor metastasis to the lungs 34 months after the presentation. To our knowledge, only 3 similar cases have been previously described in the literature.

Background: Many different methods have been used to assay amylase activity, using nitrophenylated oligosaccharides as substrate; however, the hydrolysis steps in these methods are complex.Methods: We developed a new continuously monitoring assay for amylase activity in biological fluids, using 2-chloro-4-nitrophenyl-4-O-β-d-galactopyranosylmaltoside (GalG2CNP) as the substrate; this assay was used with anti-human salivary amylase monoclonal antibodies for specific determination of the pancreatic isoenzyme. Amylase converted GalG2CNP into β-d-galactopyranosylmaltose and 2-chloro-4-nitrophenol, which was measured at 405 nm.Results: GalG2CNP was cleaved between 2-chloro-4-nitrophenol and β-d-galactopyranosylmaltose and did not undergo transfer reactions. The within-assay CVs (n = 20) for total amylase (T-AMY) and pancreatic amylase (P-AMY) were 0.6–1.6% and 0.5–2.5%, respectively; and day-to-day CVs (n = 10) for T-AMY and P-AMY were 0.8–3.7% and 0.6–4.1%, respectively. T-AMY and P-AMY activities in serum or urine obtained by the proposed method correlated well with those determined by the 2-chloro-4-nitrophenyl 4-O-β-d-galactopyranosyl-β-maltotetraoside method or the modified IFCC method.Conclusions: This novel assay for T-AMY and P-AMY measures both activities stoichiometrically, directly, and easily, and may be suitable for routine procedures.

Vasculature was examined immunohistochemically in the villous stroma in 22 cases of complete hydatidiform mole (CHM). Only antibody CD-34 was able to detect the vasculature. Serial thin and thick sections and squashed preparations of molar villi were stained by CD-34, and sequential photographs of the vasculature were taken and projected on paper, and the vasculature was described and reconstructed. In the stroma of almost all molar villi examined, vessels were found, but their density and structure varied greatly in each molar villus even in the same CHM, and the vessels were irregular in diameter. In fully distended molar villi the vessels often became narrow, continuing like a broken line but eventually vanishing. Generally the density of the vessels was inversely proportional to the grade of the swelling of villi. The vessels sometimes passed through two adjacent molar vesicles. No significant differences in vasculature were found in molar villi at different gestational ages.

Background —Transforming growth factor-β (TGF-β) is an important regulator of vascular remodeling and is involved in the pathogenesis of atherosclerosis. A T→C transition at nucleotide 29 of the TGF-β1 gene results in a Leu→Pro substitution at amino acid 10 of the signal peptide. We have now examined a possible association of TGF-β1 genotype with myocardial infarction (MI) in a Japanese population. Methods and Results — TGF-β1 genotype was determined in 315 Japanese patients (234 men and 81 women) with MI and 591 control subjects (289 men and 302 women). We found that age, body mass index, and incidence of habitual smoking, hypertension, diabetes mellitus, and hypercholesterolemia did not differ between the 2 groups for either men or women. Multivariable logistic regression analysis, however, demonstrated the frequency of the T allele to be significantly higher in male subjects with MI than in controls ( TT + TC versus CC ; P T allele was not associated with the prevalence of MI in women. In both male MI patients and controls, the serum concentration of TGF-β1 was significantly higher in individuals with the CC genotype than in subjects with the TT or TC genotype. Conclusions —Findings suggest that the T allele at nucleotide 29 in the TGF-β1 gene is a risk factor for genetic susceptibility to MI, at least in middle-aged Japanese men.

The t(X;18)(p11.2;q11.2) translocation commonly found in synovial sarcoma (SS) results in the fusion of the SYT gene on chromosome 18 to either of two closely related genes, SSX1 and SSX2, on chromosome X. It has been suggested that patients who have SS bearing SYT-SSX1 fusion have worse prognosis than those bearing SYT-SSX2 fusion. However, little is known about the biologic basis or the relationship with the histopathologic risk factors in regard to the different fusion types. We analyzed 19 cases of SS with no metastasis at diagnosis. These tumors were classified by reverse transcription-polymerase chain reaction to SYT-SSX1 and SYT-SSX2 types. The expression of Ki-67, p27, p53, and bcl-2 and various clinicopathologic parameters including mitotic rate were compared between the two fusion types. The SYT-SSX1 type fusion was associated with high Ki-67 expression (P = .011) and high mitotic rate (P = .070). No significant differences were found between the two types as to the expression of p27, p53, and bcl-2 and other clinicopathologic parameters. The survival analysis showed that SYT-SSX1-type fusion, high Ki-67 expression, and high mitotic rate correlated with shorter metastasis-free survival. These data suggested that SYT-SSX fusion type is associated with tumor cell proliferative activity and prognosis of patients who have SS.

Between January 1992 and July 1997, 202 cases of epithelial ovarian cancer were registered and assigned randomly to a combination of cisplatin and carboplatin (PP group), or cisplatin, vinblastine and bleomycin (PVB group). We analyzed 189 patients whose clinical records were available. The PP chemotherapeutic regimen was advantageous in terms of overall survival compared to the PVB regimen until 4 years after the initial operation. However, the 5-year survival rates were almost the same in both groups. However, in stage III patients, the mean survival time in the PP group was 51.4 months and that in the PVB group was 23.3 months, and there was a statistically significant difference in the survival curves between the two groups (p = 0.0158). The 5-year survival rates were 31.1 and 20.4% in the PP and PVB groups, respectively, in stage III patients. The PP regimen was also significantly superior in patients with macroscopic residual tumor after the initial operation, and the 5-year survival rates were 25.7 and 10.1% in the PP and PVB groups, respectively (p = 0.0128). However, there was no significant difference between the two regimens in patients without macroscopic residual tumor. Cox’s proportional hazards regression analysis showed that tumor stage, presence of macroscopic residual tumor, and the chemotherapeutic regimen used were significant prognostic factors. In conclusion, the PP chemotherapeutic regimen is superior to the PVB regimen especially in the treatment of advanced ovarian cancer.

Mutation of the cardiac troponin T (cTnT) gene is a genetic determinant of familial hypertrophic cardiomyopathy (HCM). A Japanese family of 14 individuals, including 6 with HCM, was subjected to genetic and clinical assessment. Five exons of the cTnT gene were sequenced in all family members. A heterozygous or homozygous T340→A (Phe110→Ile) mutation in exon 9 of the cTnT gene was detected in 11 subjects. Morphological and functional evaluation of the left and right ventricles by echocardiography revealed that 4 of 9 individuals heterozygous for the mutant allele exhibited HCM with moderate cardiac hypertrophy. Cardiac hypertrophy and other clinical features in the 2 subjects homozygous for the mutation were more severe than were those in heterozygous individuals with HCM. Thus, the clinical features of HCM due to the Phe110→Ile mutation in the cTnT gene appear to be modified by a gene dosage effect.

Objective The purpose of the present study was to determine whether electroencephalographic (EEC) and cardiac arousal, i.e. heart rate elevation at the termination of apnea/hypopnea are related to aging. Methods The subjects were 13 middle-aged (40-60 years of age, 47.8±5.35 years) and 10 elderly (>60 years of age, 69.8±5.31 years) male patients with obstructive sleep apnea syndrome. We evaluated the number of apneas per an hour of sleep (apnea index: AI), the number of hypopneas per an hour of sleep (hypopnea index: HI), the summation of HI and AI (apnea/hypopnea index: AHI), sleep stage, the amount of time during which nocturnal oxygen saturation was decreased below 90% (oxygen desaturation time: ODT), and EEG and cardiac arousal at the termination of apnea/hypopnea using polysomnography with pulse oximetry. Results There was no significant difference in AHI, duration of apnea/hypopnea, and sleep stage distribution between the two groups. However, the ratio of apnea/hypopnea with EEG and cardiac arousal was significantly higher in middle-aged than in elderly patients. The ratio of HI to AHI was significantly lower in middle-aged than in elderly patients. In middle-aged patients, EEG and cardiac arousal were significantly correlated with AI, AHI, and ODT, whereas, in elderly patients, these parameters were not similarly correlated. Conclusion Our findings suggest that the differences in EEG and cardiac arousal, and the pattern of sleep-disordered breathing (SDB) between middle-aged and elderly patients with SDB might be a physiological phenomenon of aging.(Internal Medicine 39: 375-380, 2000)

Aminopeptidase A (AP-A), a cell-surface metallopeptidase hydrolyzing peptide with N-terminal acidic residues, has been proved to be identical to the B cell differentiation antigen BP-1 and to the kidney differentiation antigen gp160, suggesting recognition of AP-A as a differentiation-related marker on certain normal and transformed cells. AP-A has also been purified from human placenta and been shown to be localized in the trophoblasts. In the present study, we examined the expression and enzymatic activity of AP-A in human gestational choriocarcinoma, a neoplastic transformant from trophoblasts which comprises a heterogenous population of trophoblastic cells in different stages of differentiation. Flow cytometry and immunoblot analysis demonstrated that AP-A was expressed in five choriocarcinoma cell lines which were secreting low or moderate levels of human chorionic gonadotropin (hCG), while two high hCG-secreting cell lines lacked AP-A expression. The AP-A enzymatic activity correlated with cell-surface levels of AP-A and was abrogated by amastatin, an inhibitor of AP-A. Immunohistochemical analysis revealed that AP-A was present in seven of eight choriocarcinoma tissues and was localized on the cell membrane of cytotrophoblastic choriocarcinoma cells, but not on cells with syncytiotrophoblast-like features. These results demonstrate that AP-A is expressed on most choriocarcinomas and its expression is restricted to low hCG-secreting, cytotrophoblastic cells and down-regulated as a function of cell differentiation, suggesting an involvement of AP-A in the differentiation/maturation process of neoplastic trophoblasts.

Aminopeptidase A (AP-A) is a cell surface metallopeptidase which specifically cleaves the amino-terminal acidic residue from peptide substrates such as angiotensin II. AP-A is identical to the differentiation-related antigen, murine BP-1 or human kidney gp160, and is involved in regulating cell differentiation and/or neoplastic transformation of certain normal and transformed cells. We examined expression of AP-A in premalignant and malignant lesions of the uterine cervix, and investigated whether its expression was related to disease progression and neoplastic transformation. Formalin-fixed, paraffin-embedded tissue sections including 14 cervical intraepithelial neoplasms (CIN) and 23 invasive squamous cell carcinomas (SCC) were immunohistochemically evaluated. AP-A was localized in the basal cell layer in normal squamous epithelium. In CIN, AP-A expression was found on dysplastic cells, and increased with the severity of the precancerous lesions. In invasive cancer, 18 of 19 non-keratinizing-type SCCs and none of 4 keratinizing-type SCCs expressed AP-A. In addition, AP-A immunoreactivity was significantly correlated with proliferating cell nuclear antigen expression in both CIN and SCC cases. Furthermore, angiotensin II type 1 receptor was present in all AP-A-positive SCCs. These results indicate that AP-A is upregulated as the lesion progresses toward carcinoma in the cervical epithelium, and suggest that AP-A may play a regulatory role in neoplastic transformation and disease progression in cervical neoplasms and may serve as a potential tumor marker during cervical neoplasia development.

To determine the outcome of patients with endometrial endometrioid adenocarcinoma following adjuvant chemotherapy, CAP (cyclophosphamide, pirarubicin and cisplatin) and EP (etoposide and cisplatin) were assigned at random to patients with Ic or more advanced stage carcinoma, and their efficacy was compared. These patients were treated by the Tokai Endometrial Cancer Study Group (Nagoya University and related institutions) between January 1992 and June 1996. The 5-year survival rate was 88.4% in the CAP group and 95.1% in the EP group; the difference between the two groups was not significant (p = 0.3496). The disease-free survival rate was 80.3% in the CAP group and 84.8% in the EP group (nonsignificant: p = 0.4533). However, the 5-year disease-free survival rates were 95.1 and 71.0% in patients with preoperative CA125 levels

Various methods for determining calcium in body fluids have been reported. Atomic absorption spectrophotometry (AAS) (1) is a most reliable method; however, because the AAS instrument is expensive and maintenance is difficult, it is unsuitable for routine assay. Spectrophotometric methods by use of o -cresolphthalein complexone (CPC) (2)(3)(4)(5)(6), which is based on chelation, is now widely available to assay calcium in clinical laboratories. Several enzymatic methods that use porcine pancreatic α-amylase (EC 3.2.1.1) (7) or phospholipase D (PLD; EC 3.1.4.4) (8) to measure calcium have also been developed. These methods are based on the activation of the enzymes by calcium. On the other hand, Kimura et al. (9) reported a kinetic assay for calcium in serum that uses urea amidolyase (EC 6.3.4.6) based on the inhibition of the enzyme by calcium. Previously reported method for determining free calcium ions in serum using PLD required appropriate ionic strength for its reaction (8). We have established a new and stable enzymatic method for determining calcium in serum that uses PLD, stabilizing calcium status by adding a large excess of bovine albumin to the reaction system. The proposed enzymatic method for determining calcium in serum is based on the following sequence of reactions: \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{Diacetylphosphatidylcholine{+}\ H}_{\mathrm{2}}\mathrm{O}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\ \begin{array}{l}\mathrm{Phospholipase\ D}\\{\rightarrow}\\\mathrm{Calcium}\end{array}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{Choline\ {+}\ diacetylphosphatidic\ acid}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{Choline\ {+}\ 2\ O}\_{\mathrm{2}}\mathrm{\ {+}\ H}\_{\mathrm{2}}\mathrm{O\ }\ \begin{array}{l}\mathrm{Choline\ oxidase}\\{\rightarrow}\end{array}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{\ Betaine\ {+}\ 2\ H}\_{\mathrm{2}}\mathrm{O}\_{\mathrm{2}}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{2\ H}\_{\mathrm{2}}\mathrm{O}\_{\mathrm{2}}\mathrm{\ 4-aminoantipyrine\ {+}\ EMSE}\ \begin{array}{l}\mathrm{Peroxidase}\\{\rightarrow}\end{array}\] \end{document} \batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \[\mathrm{\ Quinone\ dye\ (purple)}\] \end{document} Diacetylphosphatidylcholine (DAPC) is used for the PLD reaction substrate. The reaction rate at which PLD hydrolyzes DAPC into choline and diacetylphosphatidic acid depends on the amount of calcium in serum. Choline oxidase (COD) produces betaine and H2O2 from choline and O2. Peroxidase (POD) produces quinone from H2O2 with 4-aminoantipyrine (4-AA) and N -ethyl- N -(3-methylphenyl)- N ′-succinyl-ethylenediamine (EMSE). The absorbance at 546 nm for the quinone dye is measured. This proposed method and the conventional methods were performed with the Hitachi Model 7170 automated …

Object. The enzyme DNA topoisomerase IIα (Topo IIα) was tested as a measure of cell proliferation in gliomas.Methods. Immunostaining for the Topo IIα and for the Ki-67 antigen (MIB-1 antibody) was performed in paraffin-embedded tissue sections obtained from 25 resected human gliomas. Additionally, cultured human glioma cells were subjected to simultaneous flow cytometry to determine Topo IIα and DNA content.Using flow cytometric analysis, the authors found that the Topo IIα antibody labeled cells in the S, G2, and M phases of the cell cycle and also those in some parts of the G0 and G1 phases. In histological sections, Topo IIα showed more distinct staining than MIB-1, particularly in older archival cases. The proliferative indices (PIs) based on cells staining for MIB-1 and Topo IIα correlated highly with one another (r = 0.96). The Topo IIα PI immunopositivity was seen in 4.07% of cells in the low-grade astrocytoma group, 11.97% in the anaplastic astrocytoma group, and 13.84% in the glioblastoma multiforme group, representing significant differences between low-grade astrocytoma and both anaplastic astrocytoma and glioblastoma. A Topo IIα PI less than 5% predicted longer patient survival (p = 0.003).Conclusions. Immunostaining for Topo IIα represents a useful alternative to MIB-1 as a proliferative index in human gliomas.

Background: Although nocturnal pulseoximetry is routinely performed in obstructive sleep apnea syndrome (OSAS), pulseoximetry over a 24-h period has not been studied. Hypothesis: The purpose of the study was to determine whether simultaneous 24-h oxygen desaturation and electrocardiographic (ECG) recording might be used to screen for daytime sleep sequelae in patients with OSAS. Methods: Simultaneous recording of arterial oxygen saturation (SpO 2 ) and ECG was conducted over a 24-h period in 18 male patients with OSAS (mean age 51.3 years) who were diagnosed by standard polysomnography (PSG), and in 15 age-matched healthy subjects (mean age 52.7 years) as controls to evaluate circadian variation of these parameters. The measures of heart rate variability (HRV) were calculated from 24-h ambulatory ECGs. Seventeen patients with OSAS showed excessive daytime sleepiness (EDS). We calculated the duration in which SpO 2 decreased to

We developed a novel simple method to determine serum inorganic pyrophosphatase (PPase; EC 3.6.1.1) activity using purine nucleoside phosphorylase, xanthine oxidase and peroxidase, which method is applicable to automated analysis. Good reproducibility was demonstrated, with a coefficient of variation of 2.5 to 4.0% depending on the PPase activity. Good stability after color development was also observed. The presence of ascorbic acid, bilirubin, hemoglobin or intra-lipid did not influence the results. We also investigated the relationship between the activities of alkaline phosphatase (ALP; EC 3.1.3.1) and PPase. A strong positive correlation between ALP from bone and placenta and PPase was demonstrated with a correlation coefficient of 0.932 and 0.909, respectively. In contrast, ALP from liver showed a relatively weak correlation, with a correlation coefficient of 0.839. Further, placenta showed a higher ratio of PPase activity to ALP activity than did other tissues. PPase and ALP could not be distinguished by gel filtration or by ion-exchange chromatography.

子宮頸部乳頭状扁平上皮癌はまれな疾患である. 今回われわれは, 老人保健法に基づく子宮がん検診で近医を訪れ, 発見された乳頭状扁平上皮癌の症例を経験した.細胞診標本では, 出血・壊死を伴う腫瘍性背景の中に軽度異形成から扁平上皮癌までを示唆する異型細胞が多数認められ, さらに膀胱の移行上皮癌に類似した乳頭状の異型細胞集塊が認められた.生検組織診では, 乳頭状扁平上皮癌と診断されたが, 浸潤の有無は確認できなかった. 術後の組織診では浸潤が認められた.乳頭状扁平上皮癌の症例においては, 術前の生検組織診では浸潤を確認できない例が多いが, 細胞診では浸潤の推定が可能な例が多く, 細胞診は術前の病期診断に有用である.

We compared the ability of the semiautomated BDProbeTec-SDA system, which uses the strand displacement amplification (SDA) method, with that of the Roche Amplicor-PCR system and the Septi-Chek AFB culture system to directly detect Mycobacterium tuberculosis complex (MTB) and other mycobacteria in sputum samples. A total of 530 sputum samples from 299 patients were examined in this study. Of the 530 samples, 129 were culture positive for acid-fast bacilli with the Septi-Chek AFB system; 95 for MTB, 29 for M. avium-M. intracellulare complex (MAC), and 5 for other mycobacteria. The BDProbeTec-SDA system detected 90 of the 95 samples culture positive for MTB (sensitivity, 94.7%), and the Amplicor-PCR system detected 85 of the 95 samples culture positive for MTB (sensitivity, 89.5%). The specificity of each system, based on the clinical diagnosis, was 99.8% for SDA and 100% for PCR, respectively. Among the 29 samples culture positive for MAC, the BDProbeTec-SDA system detected MAC in 24 samples (sensitivity, 82.8%), whereas the Amplicor-PCR system detected MAC in 23 samples (sensitivity, 79.3%). The specificities of the systems were 98.3 and 100%, respectively. The high degrees of sensitivity and specificity of the BDProbeTec-SDA system suggest that it should be very useful in clinical laboratories for the rapid detection of mycobacteria in sputum samples.

The association of an insertion/deletion (I/D) polymorphism in the angiotensin converting enzyme (ACE) gene or the serum activity of ACE with coronary artery disease (CAD) was investigated in Japanese men and women. The ACE genotype of 947 CAD subjects who underwent coronary angiography and of 893 control subjects was determined by polymerase chain reaction analysis. No association of the DD genotype or the D allele with CAD was observed in men or women. In a low risk group (defined by a body mass index below the median value and the absence of a history of hypertension, diabetes mellitus, and hypercholesterolemia), there was also no association between the ACE gene polymorphism and CAD. No significant difference in serum ACE activity was detected between CAD subjects and controls of all genotypes or of the same genotype, whereas a significant association was apparent between serum ACE activity and ACE genotype for both CAD subjects and controls among both men and women. These results indicate that the ACE I/D polymorphism and genotype associated variation in serum ACE activity are not risk factors for CAD in Japanese men or women.

We describe a new kinetic assay for determining urea in serum or urine with use of urease (EC 3.5.1.5) and leucine dehydrogenase (EC 1.4.1.9). The latter enzyme is suitable for the kinetic assay of NH4+ because itsKmvalue for NH4+ at pH 8.75 is large (∼500 mmol/L). Interference from endogenous NH4+ in serum or urine is obviated by subtraction of the assayed endogenous NH4+value in a sample blank. For serum, within-assay CVs (n = 10) were 0.39–0.58%; day-to-day CVs (n = 10) were 1.56–2.30%. In urine, within-assay CVs (n = 10) were 0.86–1.15%. Analytical recovery of urea (0.893–71.4 mmol/L) added to patients’ sera (urea 6.14 mmol/L) was 99.2–105.2%. The calibration curve for serum was linear through zero for urea concentrations up to 142.9 mmol/L and for urine up to 714.3 mmol/L. No influences of added ammonium ion, bilirubin, hemoglobin, ascorbic acid, or Intralipid were observed. The regression equations for this method (y) and conventional methods (x = Determiner-LUN for serum assays, Serotec UUR-R for urine) were: y = 1.016x − 0.12 mmol/L (r = 0.999, Sy|x = 0.34 mmol/L, n = 100) for sera, and y = 1.070x − 12.6 mmol/L (r = 0.998, Sy|x = 7.41 mmol/L, n = 100) for urine.

1. The aim of the present study was the evaluation of human gastric ulcer healing and intractability from the viewpoint of connective tissue components. 2. Based on investigations of clinical status, we divided 78 patients with a gastric ulcer into three groups. All patients underwent gastrectomy and group I consisted of 17 patients operated on for bleeding or perforation, group II consisted of 37 patients operated on for an accompanying gastric carcinoma and group III consisted of 24 patients operated on for ulcer intractability. 3. We evaluated, in resected specimens, angiogenesis by Azan-Mallory staining and fibroplasia by applying proliferating cell nuclear antigen (PCNA) immunostaining. The inflammatory grade and fibrosis at the ulcer base were also evaluated by haematoxylin-eosin and Azan-Mallory staining. 4. Microvessel counts in group III were significantly lower than those in group I and II (P < 0.05). The PCNA labelling index of fibroblasts in group III was significantly lower than that in groups I and II (P < 0.05). The inflammatory grade was severe and fibrosis was excessive in group III. 5. From these results, it is suggested that the reduced angiogenesis and decline in fibrous cell proliferation are important factors contributing to gastric ulcer intractability.

We have compared the ability of the Mycobacterium Growth Indicator Tube (MGIT) system, a new culture method with an oxygen-sensitive fluorescent sensor, to recover mycobacteria from sputum samples with the abilities of egg-based medium and the Septi-Chek AFB system. We have also assessed the clinical utility of the AccuProbe or the AMPLICOR-PCR assay to directly identify Mycobacterium tuberculosis complex and M. avium-M. intracellulare complex (MAC) from positive MGITs. From 382 sputum samples, 99 isolates of M. tuberculosis complex and 20 isolates of MAC were recovered. The MGIT system had the highest recovery rates for M. tuberculosis complex (97.0%) and MAC (100%), compared to recovery rates of 51.5 and 65.0%, respectively, with the egg-based medium and 81.8 and 85.0%, respectively, with the Septi-Chek AFB system. The shortest recovery times were also achieved with the MGIT system: 16.6 days for M. tuberculosis complex and 12.0 days for MAC, compared to 27.1 and 20.1 days, respectively, with the egg-based medium and 21.4 and 13.2 days, respectively, with the Septi-Chek AFB system. The AccuProbe identified 74 (77.1%) of the 96 M. tuberculosis complex-positive MGITs and 17 (85.0%) of the 20 MAC-positive vials. The AMPLICOR system correctly identified 94 (97.9%) of the 96 M. tuberculosis complex-positive MGITs and all 20 MAC-positive vials. Therefore, the MGIT system used in conjunction with the AMPLICOR system is a rapid and sensitive method for detecting and identifying M. tuberculosis complex and MAC isolates from sputum samples.

The relationship of an insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene to left-ventricular hypertrophy in individuals with essential hypertension (EH) was investigated in a large population of Japanese men and women. The ACE genotype of 762 subjects with EH (425 men and 337 women) and 1,157 healthy controls (604 men and 553 women) was determined by polymerase chain reaction analysis. The distribution of ACE genotypes did not differ significantly between patients with EH and control in both men and women. For women with EH, the DD genotype was positively associated with the thickness of the interventricular septum and inversely associated with the left ventricular end-diastolic dimension, both determined by echocardiography. In contrast, the DD genotype was not associated with any echocardiographic parameter in men with EH. These results indicate that the DD genotype is a risk factor for left-ventricular hypertrophy in Japanese women with EH, but not for Japanese men.

The goal of the present investigation is to show the clinical significance of arousal response at termination of apnea/hypopnea in patients with sleep apnea syndrome (SAS) after cerebral infarction. We polygraphically assessed "cardiac arousal," which is defined as an abrupt increase in heart rate at a termination of sleep apnea/hypopnea, and electroencephalographic (EEG) arousal. There were six elderly subjects, bedridden after cerebral infarction, with SAS aged 71-87 years (mean 72.3 years) and 11 age-matched patients with SAS aged 61-78 years (mean 62.3 years) as controls. The following sleep parameters were measured: number of apneas per hour (apnea index [AI]), number of hypopneas per hour (hypopnea index [HI]), summation of the two (apnea/hypopnea index [AHI]), and duration in which nocturnal oxygen saturation was decreased below 90% (duration of SaO2 < 90%). We calculated the ratio of apnea/hypopnea per hour with cardiac arousal to total apnea/hypopneas (XI) (% cardiac arousal [XI/AHI x 100]) and the ratio of that with EEG arousal (YI) (% EEG arousal [YI/AHI x 100]). Between the two groups, we found no significant difference in body mass index, the ratio of central apnea to total apnea/hypopnea, AHI, duration of apnea/hypopnea, lowest SaO2, and duration of SaO2 < 90%. Compared with controls, % cardiac and % EEG arousals were significantly lower in patients with cerebral infarction. In contrast, the ratio of HI to AHI was significantly higher in patients with cerebral infarction than in control subjects. Our findings indicate that cardiac and EEG arousals at termination of apnea/hypopnea are significantly suppressed in elderly patients with SAS after cerebral infarction, which may provide useful information on the pathophysiology of SAS in patients with cerebrovascular disease.

Immunohistochemical localization of the large proteoglycan, PG-M/versican, was studied in 36 breast tumours, including infiltrating ductal carcinomas, benign tumours and fibrocystic diseases. The relation between the proteoglycan and the other extracellular matrix components was also investigated. In the carcinoma tissues, the interstitial elements of the ‘specific stroma’, consisting of fibroblastic cells and fine fibrils, were reactive to antibody 2B1, which specifically recognizes the large proteoglycan, PG-M/versican. In the peripheral invasive areas of infiltrating ductal carcinoma, the most intense 2B1-positive reaction was visualized in mesenchymal tissues between carcinoma cell clumps and the surrounding tissues, where hyaluronic acid could be demonstrated histochemically. The 2B1-positive elements were not reactive to antibody 6B6, which specifically recognizes small proteoglycan. In the central sclerotic areas, where antibody 6B6 was reactive, a 2B1-positive reaction was detected only in elastosis masses, which also bound antibodies to type IV collagen and laminin, and to some extent antibody raised against chondroitin 6-sulphate proteoglycan. Elastic tissues of blood vessel walls and perivascular elements became reactive to antibody 2B1 when they were involved in carcinoma invasion. The present results have shown that PG-M/versican was localized in the proliferating interstitial tissues, in particular in hyaluronic acid-rich portions, in association with carcinoma cell growth, and also that PG-M/versican accumulated in vascular and perivascular elastic tissues involved in carcinoma invasion. The biological significance of PG-M/versican was briefly discussed

Forty-seven strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated from 47 patients with bacteremia were analyzed by chromosomal DNA digestion pattern using pulsed-field gel electrophoresis and evaluated for serological coagulase type, enterotoxin type, and toxic shock syndrome toxin-1 production. The mortality rate was significantly higher in the older patients (> or = 51 years of age) than in the younger patients (< or = 50 years of age) (50% vs. 4%, p = 0.0007). Methicillin-resistant Staphylococcus aureus strains of serological coagulase type II were more likely to be associated with mortality in older patients than were strains of the other types (p = 0.037).

The intraportal endovascular ultrasonography (IPEUS) could diagnose the second portion of the extrapancreatic nerve plexus invasion and provide precise information in operative strategy. But, the first portion was not visualized clearly owing to poor tissue penetration of the ultrasound beam, which may have reduced diagnostic accuracy. Improvement of the scanning area is expected to make intraportal endovascular US even more useful.Pancreatic cancer easily invades the retroperitoneal tissue, especially the extrapancreatic nerve plexus. We evaluated the extrapancreatic nerve plexus invasion of the pancreatic cancer with IPEUS. IPEUS was performed intraoperatively in 20 consecutive resected cases with carcinoma of the head of the pancreas.IPEUS was performed with an 8-French, 20 MHz intravascular ultrasound catheter. IPEUS visualized the inferior pancreaticoduodenal artery (IPDA) in the extrapancreatic nerve plexus. The high-echoic area around the IPDA corresponds to the second portion of the extrapancreatic nerve plexus. The sonographic criterion for detection of the extrapancreatic nerve plexus invasion is low-echoic infiltration around the IPDA.Extrapancreatic nerve plexus invasion was confirmed with resected specimens in 10 patients. The IPDA could not be visualized in two patients. In 18 patients, the diagnostic accuracy of invasion was evaluated. For diagnosis of extrapancreatic nerve plexus invasion with intraportal endovascular US, the sensitivity, specificity, and overall accuracy were 87.5, 90, and 88.7%, respectively.

To clarify whether apoptosis is involved in endometriosis, we obtained eutopic endometrial tissues along with endometriotic tissues from the uterus (adenomyosis) (n = 12) and from the ovary (n = 12) from patients undergoing gynaecological surgery. Apoptosis-induced DNA fragmentation was detected by the TdT-mediated dUTP-biotin nick-end labelling method, and immunostaining with a monoclonal antibody against the Fas, Le(y) or B-cell leukaemia/lymphoma-2 (bcl-2) was also performed using the same tissue section. Analysis showed that apoptosis was occurring in all the samples of ovarian endometriotic tissue but in only two of the 12 adenomyotic and in five of the 24 eutopic endometrial tissue samples. In none of these cases was apoptosis correlated with phases of the menstrual cycle. The expression of bcl-2 in the eutopic endometrial and adenomyotic tissues was limited to the proliferative phase, and was observed in only one of the 12 cases of ovarian endometriosis. Fas and Ley were expressed randomly across a wide range in both the eutopic and ectopic endometrial tissues. These results suggest that the features of ovarian endometriosis are different from those of adenomyosis and eutopic endometrium in terms of the involvement of apoptosis. In addition, the regulatory mechanism involved in ovarian endometriosis may differ from that in other endometrial cells.

Since the incidence of mucinous carcinoma of the ovary is relatively low, with only small numbers of cases at any institution, detailed clinicopathologic studies on the prognosis and the care of patie

OBJECTIVE: To solve the problem of diagnosing apocrine carcinoma (APCA) through distinguishing it from benign apocrine metaplasia with atypia (APMA). STUDY DESIGN: The study group consisted of five histologically confirmed cases of uncommon infiltrating apocrine carcinoma and a case of noninfiltrating apocrine carcinoma of the breast by aspiration biopsy cytology. The control group consisted of 103 cases of benign apocrine metaplasia with no atypia (APMN), 4 cases of APMA and 34 cases of common-type adenocarcinoma that were encountered in 662 breast aspirations from 1988 to 1992 at Hekinan Municipal Hospital. RESULTS: In APCA the average age of patients (65±17.7 SD) (mean ± SD) was more than 20 years older than APMA, and APCA generally measured ≥2 cm or more in diameter as compared to ≤2 cm in APMA lesions. In APCA the apocrine cells had high cellularity, with the cells occur singly and with syncytia in the background. Numerous degenerated apocrine cells and characteristic cell detritus were found. APCA was also characterized by apocrine cells with more marked nuclear abnormalities, including hyperchromasia and irregular nuclear shape, frequently with irregular nucleoli; more nuclei measure ≥12 μm in diameter than in APMA. These findings, however, were present only to a mild degree in APMA, if at all. APMA may coexist with APCA. CONCLUSION: If APMA is diagnosed, an open biopsy should be performed to distinguish it from APCA.

We examined the role of intracellular calcium release in the regulation of CRH-induced ACTH secretion using the AtT20 corticotroph cell line. We found that ruthenium red, an inhibitor of ryanodine receptor, substantially diminished the secretory response, whereas Xestospongin C, an inositol 1,4,5-triphosphate receptor antagonist, had no effect. Expression of two ryanodine receptor subtypes (RyR1 and RyR3) was confirmed by RT-PCR. We also found that caffeine, a ryanodine receptor agonist, significantly stimulated, whereas thapsigargin, which causes depletion of intracellular calcium store, markedly diminished, the ACTH release. These results suggest that ryanodine receptor-mediated calcium-induced calcium release is involved in the regulation of CRH-induced ACTH release.