Quantitative differences in telomerase activity among malignant, premalignant, and benign ovarian lesions.

Telomerase activation has been demonstrated in both cancers and some noncancerous lesions. However, few studies have determined levels of telomerase activity in these lesions. In the present study, using a recently developed stretch PCR assay, telomerase activity was quantitatively determined in a variety of ovarian lesions including 36 ovarian cancers, 5 ovarian low potential malignancy (LPM) lesions, 10 ovarian cysts, and 12 normal ovaries. Telomerase activity was normalized to control activity (100 units) in C33A cell line and given in relative units. Telomerase activity in ovarian cancer (51 +/- 7 units, mean +/- SE) was significantly higher than that in LPM lesions, ovarian cysts, and normal ovaries (7 +/- 3, 10 +/- 2, and 10 +/- 2 units, respectively; P < 0.001). Interestingly, all LPMs, ovarian cysts, and normal ovaries exhibited low telomerase activity less than 30 units, and no significant difference in level of telomerase activity was found among them. We also found a significant correlation between the level of telomerase activity and the clinical stage of ovarian cancer. Our quantitative telomerase assay thus clearly distinguished telomerase activity in ovarian cancers from that in LPM lesions and ovarian cysts. Significant levels of telomerase activation frequently occurred in cancer but rarely occurred in premalignant and benign lesions, suggesting that telomerase activation is a critical step in cancer development.

Aromatase and 17 beta-hydroxysteroid dehydrogenase type 1 in human breast carcinoma.

The in situ formation of estradiol plays an important role in the development and biological behavior of human breast cancer Aromatase and 17 beta-hydroxysteroid dehydrogenase type 1 (17 beta-HSD type 1) are two principal enzymes involved in in situ estradiol production. We evaluated the expression of aromatase and 17 beta-HSD type 1 by immunohistochemistry in 41 cases of invasive breast carcinoma (19 lobular and 22 ductal). We then examined the correlation among the expression of these enzymes, estrogen (ER) and progesterone (PR) receptor status, Ki67 labeling index of carcinoma cells, age, and the clinical stage of the patients. Marked aromatase immunoreactivity was observed in stromal cells around carcinomatous glands in 32 of 41 cases (78%), and 17 beta-HSD type 1 immunoreactivity was detected in carcinoma cells in 23 of 41 cases (56%). There was a significant correlation observed between expression of 17 beta-HSD type 1 and aromatase in invasive lobular carcinoma (P = 0.0119), but not in invasive ductal carcinoma. There was an inverse correlation between aromatase and ER status in invasive ductal carcinoma (P = 0.0213), but not in invasive lobular carcinoma. No other correlations were observed among 17 beta-HSD type 1, aromatase, PR, ER, clinical stage, age, and Ki67 labeling indexes. Aromatase and 17 beta-HSD are not always expressed simultaneously in human breast carcinoma, but their simultaneous expression is more frequent in invasive lobular carcinoma than invasive ductal carcinoma. Consequently, different mechanisms may be involved in the regulation of expression of these two enzymes in human breast carcinoma.

Elevated antibody-dependent cell-mediated cytotoxicity and its inhibition by nicotinamide in the diabetic NOD mouse.

Antibody-dependent cell-mediated cytotoxicity (ADCC) by splenic mononuclear cells was measured in female non-obese diabetic (NOD) mice and age-matched ICR mice. No significant difference in ADCC activities was observed between the two groups when all the NOD mice were pre-diabetic. ADCC activities in diabetic NOD mice were significantly higher than those in age-matched ICR mice (P less than 0.001). Nicotinamide, known to prevent the diabetes of the NOD mouse, strongly inhibited ADCC by the mononuclear cells from diabetic NOD mice. Kinetic studies revealed that the inhibition was non-competitive.

Effects of cyclopiazonic acid on contraction and intracellular Ca2+ in oesophageal striated muscle of normotensive and spontaneously hypertensive rats.

1. The effects of cyclopiazonic acid (CPA), a selective inhibitor of sarcoplasmic reticulum (SR) Ca2+-ATPase, on twitch contraction and on the resting state of tension and intracellular Ca2+ level ([Ca2+]i) of the oesophageal striated muscle of stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar Kyoto rats (WKY) were compared. 2. CPA (10 micronM) augmented the twitch contraction of oesophageal striated muscle preparations from both SHRSP and WKY, reducing the rate of relaxation (-dT/dt), and thus resulting in the prolongation of the time to 80% relaxation. The effect was significantly smaller in the SHRSP preparations. 3. In the resting state, CPA caused a sustained elevation of [Ca2+]i. The elevation was greater in the WKY preparations. Tension development accompanied by the elevation was observed in WKY preparations, but not in SHRSP preparations. 4. The sustained elevation of [Ca2+]i induced by CPA was eliminated by the removal of extracellular Ca2+. Both the elevated [Ca2+]i and tension in the preparations from WKY were reduced by flufenamic acid (100 micronM), mefenamic acid (100 micronM), lanthanum (La3+, 100 micronM), gadolinium (Gd3+, 100 micronM) and SK&F 96365 (100 micronM) but not by verapamil (10 micronM). 5. Thapsigargin (3 micronM), another SR Ca2+-ATPase inhibitor, produced similar effects on basal tension to those of CPA, although it reduced the amplitude of twitch contraction. 6. These results suggest that in the rat oesophageal striated muscle, (1) CPA extends the sequestrating time of Ca2+ into the SR, (2) CPA induces a Ca2+ influx mediated through verapamil-insensitive pathways, possibly nonselective cation channels, and (3) the mechanism of [Ca2+](i) modulation due to CPA-sensitive SR Ca2+-ATPase is deteriorated in the oesophageal striated muscle from SHRSP as compared with WKY preparations.

A phase I study of weekly docetaxel and cisplatin in advanced non-small cell lung cancer.

This phase I study was designed to determine the maximum tolerated dose (MTD) and toxicity of a weekly docetaxel (TXT) and cisplatin (CDDP) combination regimen in advanced non-small cell lung cancer (NSCLC). Patients with stage IIIB or IV NSCLC who were previously untreated were eligible. Docetaxel, at a starting dose of 20 mg m(-2) per week on days 1, 8 and 15, was combined with a fixed dose of cisplatin 80 mg m(-2) on day 1. Docetaxel was increased in 5 mg m(-2) per week steps. Chemotherapy was given in a 4-weeks cycle. Dose-limiting toxicities (DLTs) were defined as grade 3-4 leukopenia, thrombocytopenia, anemia, fever with grade 4 neutropenia and more than grade 2 non-hematologic toxicity, with the exception of nausea, vomiting, and alopecia. Omission of chemotherapy on day 8 and/or 15 was also considered DLT. Eighteen patients were enrolled in this study. Leukopenia, anemia and fatigue were the DLTs. No grade 4 toxicities were seen in any patients. The overall response rate was 44.4% (95% confidence interval, 21.5-67.4%). The recommended dose of TXT to be combined with CDDP 80 mg m(-2) on day 1 is 35 mg m(-2) per week on days 1, 8 and 15. This is a promising regimen, therefore a multicenter phase II study is now under way.

Different effects of D-glucose anomers on enhanced secretion of gastric acid in rat.

The injection of alpha-, beta-, or equilibrated (alpha: 36%, beta: 64%) D-glucose solution into the cranial side of the carotid artery decreased gastric acid output caused by insulin in rats with bilateral adrenalectomy. This effect was not reproduced after vagotomy at the cervical level. Of the 3 forms of D-glucose solution the effect of beta-D-glucose was greatest. The injection of isotonic NaCl solution, however, produced no change in acid output. These results suggest that blood beta-D-glucose may play a predominant role in activating a brain mechanism which controls gastric acid secretion via the vagus nerve.

Weekly CODE chemotherapy with recombinant human granulocyte colony-stimulating factor for relapsed or refractory small cell lung cancer.

We used cisplatin, vincristine, doxorubicin, and etoposide (CODE) plus recombinant human granulocyte colony-stimulating factor (rhG-CSF) weekly for salvage chemotherapy in relapsed or refractory small cell lung cancer (SCLC). We reviewed the medical charts of patients between January 1993 and December 1996 at the National Nishi-Gunma Hospital. Twenty patients were treated with salvage chemotherapy. The overall response rate was 55.0%. The median survival time of extensive disease patients from the start of CODE therapy was 23 weeks and the 1-year survival rate was 21.0%. Toxicities were severe, especially in myelosuppression. CODE could be selected as a salvage therapy for chemotherapy- relapsed SCLC cases.

The inhibitory effect of forskolin on antibody-dependent cell-mediated cytotoxicity using Chang liver cells as target cells.

The effect of forskolin, a unique adenylate cyclase activator, on antibody-dependent cell-mediated cytotoxicity (ADCC) was examined. ADCC was assayed using Chang liver cells as the target cells, immuned rabbit serum as the antibody and healthy human peripheral blood mononuclear cells (PBMNC) as the effector cells. Forskolin at concentrations ranging from 1 to 20 microM significantly inhibited ADCC in a dose-dependent manner. By the addition of forskolin, cyclic AMP levels did not change in Chang liver cells but increased in PBMNC. Therefore, it appears that forskolin exerted an inhibitory effect on ADCC by increasing the intracellular cyclic AMP levels in PBMNC, the effector cells.

Localization of mineralocorticoid receptor and 11 beta-hydroxysteroid dehydrogenase type II in human breast and its disorders.

Mineralocorticoid receptors have been detected in the normal human breast and breast cancers. The expression of mineralocorticoid receptor (MR) and 11 beta-hydroxysteroid dehydrogenase type II (11sHSD2), which confers specificity on MR for aldosterone, was examined by immunohistochemistry in 114 samples from normal human breast and benign and malignant breast lesions in order to study its possible biological significance. MR and 11sHSD2 were immunolocalized in the ductal epithelium in 39/40 (98%) and 36/40 cases (90%) of normal breast, 21/22 (95%) and 15/22 cases (68%) of fibrocystic changes, and 11/11 (100%) and 8/11 (73%) cases of fibroadenoma, respectively. Cases positive for 11 sHSD2 also expressed MR but the patterns of expression varied greatly among examples of normal breast and benign breast diseases. There was a significant correlation between labeling indices of MR and 11sHSD2 in normal breast (p < 0.01) and in benign breast disease (fibrocystic change (p < 0.05) and fibroadenoma (p < 0.05)). In invasive carcinomas, immunoreactivity for MR and 11sHSD2 was detected in malignant cells in 32/41(78%) and 16/41(39%) cases. Both MR and 11sHSD2 labeling indices were significantly higher in invasive ductal carcinoma (22 cases) than invasive lobular carcinoma (19 cases) (p < 0.01). There was a significant correlation between labeling indices of MR and 11sHSD2 when analyzing all infiltrating carcinomas (p < 0.01), but not when assessing invasive lobular or invasive ductal carcinomas separately. These results indicate that the 11 sHSD2 enzyme generally colocalizes with the MR in the ductal epithelial cells of human breast, which may allow aldosterone to occupy its physiological receptor, and the expression of MR and 11sHSD2 appears to be related to ductal differentiation of breast carcinomas.

Immunolocalization of cyclins D and E and cyclin dependent kinase (cdk) 2 and 4 in human breast carcinoma.

We studied the immunolocalization of cyclins D1 and E and their corresponding partner cyclin dependent kinases (cdk), cdk4 and cdk2 in 41 cases of human breast malignancy (21 invasive ductal carcinomas and 19 invasive lobular carcinomas) and examined the correlation of the labeling indexes among these cyclins, cdks, Ki67, estrogen receptor (ER) and progesterone receptor (PR). Cyclin D1 immunoreactivity was observed exclusively in the nuclei of tumor cells in 27/41 (65%) of the cases examined. Immunoreactivity for cyclin E and cdk2 was detected in all the cases and observed in the nuclei of both carcinoma and non-carcinoma cells. cdk4 immunoreactivity was detected in 39/41 (95%) cases and found in carcinoma and non-carcinoma cells. In all carcinomas examined, a significant correlation was observed only between Ki67 and cyclin D1 (p = 0.0037). However, when examining only invasive ductal carcinomas, a significant correlation was detected between Ki67 and cyclin D1 (p = 0.0069), Ki67 and cdk2 (p = 0.0043) and cyclin D1 and cdk4 (P = 0.0024). Only cyclin D1 correlated with the pathologic stages of the disease and histological grades of invasive ductal carcinoma. Among these cyclins and cdk, overexpression of cyclin D1 is considered to play an important role in the development of human breast malignancy through abnormal proliferation. No significant correlation was observed between steroid receptor status and any of cyclins and cdks examined. Cyclin D1 and cdk2 expression correlated with cell proliferation (Ki67) and cyclin D1 expression with expression of cdk4 in invasive ductal carcinoma but not invasive lobular carcinoma. Cyclin E expression did not correlate with cell proliferation, cyclin D1 or cdks possibly due to deregulation of its expression. These results also indicate different patterns of cyclin D1, cyclin E, cdk2 and cdk4 expression between invasive ductal and lobular carcinoma of human breast.

Elimination rate of fat emulsion particles from plasma in Japanese subjects as determined by a triglyceride clamp technique.

The elimination rate of emulsion triglyceride (TG) from plasma was investigated in Japanese subjects by a plasma TG clamp technique. Two different studies were performed. In Study 1, a lipid emulsion (20% long-chain triglyceride emulsion: LCT) was infused into a healthy research associate to achieve a certain concentration of TG in the plasma. Thereafter, the infusion rate was adjusted to maintain the chosen concentrations of TG in plasma (namely, 4-5 mmol/L, 3-4 mmol/L, and approximately 2 mmol/L) over a period of 160 min by measuring plasma TG concentrations at 10-min intervals. Concentrations of TG in plasma were clamped within 2.13 +/- 0.13 mmol/L by an infusion rate of 0.10 g.kg-1.h-1, within 3.34 +/- 0.20 mmol/L by an infusion rate of 0.14 g.kg-1.h-1, and within 4.46 +/- 0.22 mmol/L by an infusion rate of 0.11 g.kg-1.h-1. The mean rate of infusion of emulsified TG that had maintained the steady concentrations of TG in plasma was limited to the very narrow range of 0.12 +/- 0.02 g of TG.kg-1.h-1 regardless of the chosen concentration of TG in plasma. Concentrations of nonesterified fatty acids (NEFA) also remained at a fixed level of 1.378 +/- 0.103 mEq/L regardless of the chosen concentration of TG in the plasma. Study 2 was undertaken to determine whether plasma TG concentration reached a plateau during a period when emulsion TG was infused into three different subjects at a rate of 0.12 g.kg-1.h-1. The plasma TG concentrations were steady at a level of 2.04 +/- 0.32 mmol/L, and the plasma NEFA concentrations remained at a fixed level of 1.33 +/- 0.13 mEq/L, over a period of 160 min after 50-min priming infusion. These results indicate that the plasma TG elimination rate was limited to the narrow range of 0.12 +/- 0.02 g.kg-1.h-1 when the fat emulsion was infused into Japanese subjects in a steady state. However, the plasma TG elimination rate in Japanese subjects appeared to be lower than that of Europeans. This may be due to a difference in lipoprotein lipase activity caused by different dietary habits, namely, a lower fat intake.

A phase II study of carboplatin-cisplatin-etoposide combination chemotherapy in advanced non-small-cell lung cancer.

It is reported that the combination of cisplatin (CDDP) and carboplatin (CBDCA) is synergistic in vitro. The objective of this study was to evaluate the therapeutic effect and safety of the two platinum compounds in combination with etoposide in the treatment of non-small-cell lung cancer (NSLC). Forty patients were registered. Based on the results of a phase I study, patients were treated with CDDP (80 mg/m2 i.v. on day 1), CBDCA (280 mg/m2 i.v. on day 1), and etoposide (80 mg/m2 i.v. on days 1-3). Of the 40 patients, 30 were men and 10 women. Histology revealed adenocarcinoma(AC) (n = 20), squamous cell carcinoma(SCC) (n = 18), and large cell carcinoma(LCC) (n = 2). Staging: IIIA (n = 3); IIIB (n = 17); and IV (n = 20). A 32.5% overall response rate [13 of 40; 95% confidence interval (CI) 18-47%] was achieved. The response rates in patients with SCC and AC were 55.6 and 10.0% (p < 0.005), respectively. The median duration of response was 47.1 weeks and the overall median survival time was 57.1 weeks. Leukopenia and thrombocytopenia--World Health Organization (WHO) grade IV--occurred in nine and 11 patients, respectively. Nonhematological toxicities were mainly nausea, vomiting, and alopecia. In conclusion, further investigations of this regimen are warranted in the treatment of NSLC.

Embolization for ruptured superior mesenteric artery aneurysms.

Superior mesenteric artery (SMA) aneurysms are very uncommon. They are difficult to detect until they rupture and cause hypovolaemic shock. We performed embolization in four cases of aneurysm of branches of the superior mesenteric artery, succeeding in three cases without the need for surgical treatment. In the first case, the aneurysm was excised because of migration of a microcoil into the left hepatic artery. It was not retrieved because sufficient blood flow to the liver was shown on angiography after migration and no ischaemic change of liver was detected on laparotomy. In the second case, the aneurysm arose from the anterior pancreaticoduodenal artery. In the third case, the patient had two SMA aneurysms; one had been resected at surgery, another was revealed on follow-up angiography and embolized with microcoils. The fourth patient had a jejunal artery aneurysm with extravasation; haemostasis was achieved by packing it. In all four cases, no major complications were observed in the clinical course after embolization. Microcoils were considered to be the desirable embolic material, in order to prevent post-therapeutic ischaemic change. Embolization should be the treatment of choice for SMA aneurysms, because it is less invasive and takes less time than surgical treatment.

Effect of high doses of synthetic estrogen on lipid metabolism in castrated male rats.

The mechanism by which high doses of estrogen influences lipid metabolism was studied with a microtubular blocking agent. Castrated male rats received oral injection daily for 14 days of 3 mg hexestrol in olive oil, or oil alone as controls. About half of the animals in each group were injected intraperitoneally with 4 mg/100 g body weight colchicine 3 hr before they were killed. Hexestrol treatment caused an accumulation of esterified cholesterol in the liver while it decreased those in serum. Triglyceride concentrations slightly decreased in the liver but were unaffected in serum. On polyacrylamide-gel disc electrophoresis, the peaks of high density lipoproteins (HDL) and low density lipoproteins (LDL) were decreased remarkably. Electron microscopic examination of hepatocytes revealed electron-lucent lipid droplets in the cytoplasm. After a colchicine treatment of the control animals, concentrations of esterified cholesterol and triglycerides markedly increased in the liver, while those in serum decreased. Electron microscopic examination of hepatocytes revealed numerous secretory vesicles filled with nascent VLDL. In hexestrol-treated animals, the colchicine treatment was associated with marked decreases in serum-esterified cholesterol and triglyceride as seen in the controls. However, there were no further increases of esterified cholesterol in the liver, and the increase of triglycerides was slight. Electron microscopic examination showed less secretory droplets than in the controls. These data suggest that very low density lipoproteins (VLDL) synthesis in the liver of hexestrol treated rats was inhibited. An accumulation of esterified cholesterol with a marked decrease in serum could not be accounted for by the inhibition of lipoproteins secretion, but rather by their enhanced entry into the liver.

von Recklinghausen's disease with lung cancer derived from the wall of emphysematous bullae.

A 53-year-old male with von Recklinghausen's disease, diagnosed to have metastatic large cell carcinoma in the left supraclavicular lymph node, was admitted for radiotherapy. No other primary lesions were found. Emphysematous bullae in both upper lung fields were seen on chest roentgenograph, and the leucocyte count and G-CSF level were extremely elevated. Later, when metastases appeared, systemic chemotherapy was supplemented, but in vain. An autopsy showed large cell carcinoma of the lung derived from the wall of emphysematous bullae in left lung along with multiple metastases. The presence of von Recklinghausen's disease, emphysematous bullae and lung cancer together are noteworthy.

The value of dynamic MR imaging for hypointensity lesions of the peripheral zone of the prostate.

The aim was to evaluate the role of dynamic magnetic resonance (MR) imaging for prostatic carcinoma. Forty-two men with clinical suspicion of a prostatic carcinoma underwent MR imaging. Dynamic MR was performed, followed by postcontrast T1-weighted imaging with fat suppression. Histologic diagnosis was 21 prostatic carcinomas (in 19 patients), 21 benign tissues, and 2 chronic prostatitis. The diagnostic accuracy was 75% for T2-weighted images, and 79% for dynamic images. The accuracy of the combination of dynamic MR images with postcontrast T1-weighted images was 82%. It was concluded that dynamic MR imaging was useful in differentiation of low intensity lesions in the peripheral zone.

Real-time high resolution ultrasonography of solid breast masses: use of a 10-MHz mechanical sector transducer with a water bag.

We evaluated the diagnostic accuracy of real-time high-resolution ultrasonography (RHUS) for solid breast masses, using a 10-MHz mechanical sector transducer with a water bag. A total of 103 breast nodules in 100 women were analyzed. The nodules consisted of 42 carcinomas, 10 fibroadenomas, 38 mastopathies, and 13 miscellaneous lesions. Of them, 72.8% were less than 2 cm in size. Diagnostic accuracy was correlated with the size and pathology of the nodules. Sensitivity, specificity, and accuracy were 95.2%, 83.6%, and 88.3% for carcinomas, 90.0%, 88.2%, and 88.3% for fibroadenomas, and 50.0%, 98.5%, and 80.6% for mastopathies, respectively. The overall rate of diagnostic accuracy was 74.8%. The rate of accuracy according to mass size were 87.3% (-1 cm), 87.2% (1-2 cm), 88.0% (2-5 cm) and 100% (5 cm-). False positive cases (n = 10) included seven mastopathies, two granulomas, and one fibroadenoma. False negative cases (n = 2) included one fibroadenoma and one abscess. RHUS, using a 10-MHz mechanical sector transducer, was an excellent method for the diagnosis of solid breast masses.

MR imaging of hepatosplenic sarcoidosis.

We present a case of hepatosplenic sarcoidosis. Splenic lesions were detectable by both MRI and dynamic CT. They were demonstrated as multiple low signal intensity lesions on T2-weighted images obtained by MRI and as low density lesions by dynamic enhanced CT. Hepatic focal lesions were not apparent on CT or MRI. After steroid therapy, both dynamic enhanced CT and MRI showed disappearance of the splenic lesions. These are valuable modalities for detecting focal splenic sarcoidosis and evaluating the effect of steroid therapy.

Successful treatment of idiopathic retroperitoneal fibrosis with steroid administration in a patient with a positive gallium scan.

A patient with idiopathic retroperitoneal fibrosis with a positive Ga-67 scan was successfully treated with steroids. Administration of methylprednisolone improved the ureteral obstruction and gallium accumulation also disappeared.

[A case of asymptomatic extra-adrenal pheochromocytoma].

A 46-year-old female first presented to our clinic with a chief complaint of an abdominal mass. A computerized tomographic (CT) scan revealed a 6 cm retroperitoneal mass below the left kidney. Preoperative hormonal examinations showed elevation of catecholamines and vanillymandelic acid (VMA) in her 24-hour urine, although she had no hypertension. Therefore, the tumor was uneventfully removed and histologically diagnosed as pheochromocytoma. It is difficult to preoperatively diagnose normotensive extra-adrenal pheochromocytoma without any considerations of the possibilities. However, it may be hazardous to treat pheochromocytoma surgically without any preparations. In the present case, we emphasized the importance of preoperative accurate diagnosis of extra-adrenal pheochromocytoma and discussed briefly the clinical features in the literature.