Hisako Yamamura

Mice lacking smooth muscle calponin display increased bone formation that is associated with enhancement of bone morphogenetic protein responses

Calponin is a calmodulin‐and actin‐binding protein expressed in smooth muscle. It promotes actin polymerization and inhibits actin‐activated myosin ATPase activity. Despite the molecular and functional characterization of calponin in vitro, the physiological role of calponin in vivo has not been clarified.

Values of carcinoembryonic antigen, elastase 1, and carbohydrate antigen determinant in aspirated pancreatic cystic fluid in the diagnosis of cysts of the pancreas

The levels of carcinoembryonic antigen (CEA), elastase 1, and carbohydrate antigen determinant (CA 19‐9) in the pancreatic cystic fluid and the serum from five patients with cystadenocarcinoma of the pancreas, one patient with retention cyst due to pancreatic carcinoma, three patients with cystadenoma, and eight patients with benign pseudocyst accompanying or following pancreatitis, were determined by immunoassay technique. Fluid from pancreatic cysts was obtained by ultrasonically‐guided percutaneous fine‐needle aspiration biopsy. The specimens were centrifuged and the supernatant was used for the measurement of CEA, elastase 1, and CA 19‐9, while the cell pellet was examined cytologically. The levels of CEA in the aspirated fluid were significantly higher in patients with malignant cysts of the pancreas than in those with benign cystadenomas and pseudocysts. In contrast, the levels of elastase 1 were significantly lower in patients with malignant cysts than in those with benign pancreatic cysts. Although the levels of CA 19‐9 were significantly higher in patients with malignant cysts than in those with pseudocysts, the overlap between the values of patients with malignant and benign pancreatic cysts is too great. The serum CA 19‐9 was most useful, however, to distinguish an individual patient with malignant cysts of the pancreas from those with benign pancreatic cyst, since there were no significant differences between the levels of serum CEA and elastase 1 in patients with malignant and benign pancreatic cysts. Correct diagnoses were made cytologically in 4 (66.7%) of 6 patients with malignant cysts. In two patients with malignant cyst, in whom no cancer cells were detectable in the aspirated materials, levels of CEA were abnormally high, but high levels of elastase 1 did not occur. Therefore, the combined measurement of CEA and elastase 1 in the aspirated cystic fluid of the pancreas could be used as an aid in diagnosis of malignant cysts of the pancreas.

Expression of the smooth muscle calponin gene in human osteosarcoma and its possible association with prognosis

The basic calponin gene is a smooth muscle differentiation‐specific gene that encodes an actin‐binding protein involved in the regulation of smooth muscle contractility. We studied the expression of the calponin gene in 8 human osteosarcoma cell lines and 17 primary human osteosarcoma tissues by RT‐PCR analysis. We also analyzed mRNA expression of smooth muscle‐specific genes including SM22α, caldesmon and α‐actin, and for neutral and acidic calponin isoforms. The genes were expressed at various levels by osteosarcoma cell lines and tissues of diverse histological subtypes. The basic calponin protein of an expected size was detected in osteosarcoma cell lines by immunoblot analysis and was localized by immunohistochemistry in the cytoplasm of the tumor cells in osteosarcoma tissues. Survival was found to be significantly increased in patients whose tumors exhibited basic calponin expression, compared with those with no expression. Alterations in the expression of other markers examined were not correlated with prognosis. Our results suggest that the basic calponin gene product may be a novel prognostic variable in patients with osteosarcoma. Int. J. Cancer (Pred. Oncol.) 79:245–250, 1998.© 1998 Wiley‐Liss, Inc.

Values of CA 19‐9 in the serum, pure pancreatic juice, and aspirated pancreatic material in the diagnosis of malignant pancreatic tumor

The diagnostic accuracy of the measurement of CA 19‐9 in the serum, pure pancreatic juice, and aspirated pancreatic fluid in the diagnosis of pancreatic tumors was assessed in 32 patients with malignant pancreatic tumors and 19 patients with pancreatitis. Pure pancreatic juice was collected from the pancreatic duct by endoscopic cannulation with a duodenofiberscope after intravenous administration of secretin. Pancreatic material was obtained by percutaneous fine‐needle aspiration biopsy under ultrasonic guidance. Abnormally high CA 19‐9 levels in the serum were significantly more frequent in patients with malignant pancreatic tumors than in those with pancreatitis: they were elevated in 71.9% of the patients with pancreatic tumors. High CA 19‐9 levels were found primarily in patients with a tumor of the head of the pancreas, in those with a tumor greater than 3 cm in its greatest diameter, and in those with an unresectable cancer. Only 57.1% of seven patients with a tumor of less than 3 cm in its greatest diameter showed an increase in CA 19‐9 level. The CA 19‐9 levels in pure pancreatic juice were significantly higher in patients with pancreatic tumors than in patients with pancreatitis without pancreatic stone. However, it was not useful for differentiating pancreatic tumors from pancreatitis with pancreatolithiasis. The CA 19‐9 level in pancreatic materials obtained by aspiration biopsy was significantly higher in patients with malignant pancreatic tumors than in those with pancreatitis. Eight patients (80%) with pancreatic tumors had values above 1000 U/ml, whereas all five patients with pancreatitis had values of less than 30 U/ml. Although CA 19‐9 levels in pancreatic materials was useful only when cytologic examination did not provide any evidence of malignancy, the combination of the CA 19‐9 assay and the cytologic study of specimens obtained by percutaneous fine‐needle aspiration biopsy of the pancreas increased the diagnostic rate to 100%.

Effect of Fundusectomy on Serum and Antral Gastrin Levels in Rats

In adult male rats, fundusectomy decreased acid secretion but significantly increased total antral gastrin and both fasting and food-stimulated serum gastrin levels. The rise in fasting serum gastrin could be inhibited by antral acidification, suggesting that decreased acidity caused postfundusectomy hypergastrinemia. The mechanism for the increase in total gastrin in antral tissue is probably the same. These studies provide a useful experimental model for the increasing of antral gastrin and for the production of hypergastrinemia.

Induction of calponin-h1 by transforming growth factor-β1 in cultured human Ito cells, LI90

We investigated the effect of transforming growth factor-beta1 (TGF-beta1) on the expression of calponin-h1, alpha-smooth muscle actin (alpha-SMA), and extracellular matrix (ECM) components in a cultured human Ito cell line, LI90. The TGF-beta1 treatment stimulated productions of hyaluronic acid and laminin, and significantly decreased the secretion of hepatocyte growth factor in LI90 cells. The functional characteristics of LI90 cells were compatible with those of human-activated Ito cells that are known as pericyte-like mesenchymal liver cells. TGF-beta1 induced a slight growth-inhibition of LI90 cells. TGF-beta1 enhanced the expressions of both alpha-SMA and calponin-h1 at the protein level, while tumor necrosis factor-alpha and interleukin-1alpha did not affect the expressions of these cytoskeletal proteins on LI90 cells. The addition of TGF-beta1 to LI90 cells resulted in a significant increase of calponin-h1 mRNA levels, but not calponin-h2. These data suggest that the expression of calponin-h1 is controlled at the level of mRNA under the coordinate regulation together with alpha-SMA as the process of perpetuation of activated Ito cells promoted by TGF-beta1. The identification of smooth muscle features promoted by TGF-beta1 support the hypothesis that the activation of Ito cells coincides with their contractile behavior, indicating that these cells may be important in vasoregulation during liver injury and fibrosis.

Cytologic examination and CEA measurement in aspirated pancreatic material collected by percutaneous fine‐needle aspiration biopsy under ultrasonic guidance for the diagnosis of pancreatic carcinoma

Diagnostic accuracy of the cytologic examination or CEA measurement in pancreatic fluid obtained by percutaneous fine‐needle aspiration biopsy under ultrasonic guidance, was investigated in 26 patients with histologically proven carcinoma of the pancreas, and the incidence of positive results of cytology and CEA assay were compared in pancreatic fluid obtained by percutaneous fine‐needle aspiration biopsy and by endoscopic cannulation of the ampulla of Vater in the same 19 patients. Positive cytologic findings were observed in the fluid obtained by percutaneous aspiration biopsy of 88.5% of the patients with pancreatic carcinoma. The location of the cancer had no influence on the cytology, but positive results were more frequent in patients with distant metastases than in those with localized tumor or localy invasive carcinoma. Positive cytologic results were obtained in the pancreatic fluid obtained by endoscopic cannulation from the ampulla of Vater of only 15.8% of the patients examined by both methods, but percutaneous fine‐needle aspiration biopsy significantly increased the diagnostic rate. In the specimens obtained by aspiration biopsy, tumor cells were much more abundant and easily recognizable. When cytologic examination does not provide any evidence of malignancy, measurement of CEA levels in pancreatic fluid is probably useful. Combination of the cytology and CEA assay of the specimens obtained by percutaneous fine‐needle aspiration biopsy of the pancreas increased the diagnostic rate to 100%.

Carcinoembryonic antigen in gastric juice as an aid in diagnosis of early gastric cancer

The levels of carcinoembyronic antigen (CEA) in the serum and gastric juice of 58 patients with gastric cancer were determined by radioimmunoassay. The level of serum CEA was not a satisfactory indicator of gastric cancer: it was elevated in only 19.4% of the cases of advanced and unresectable cancer and 4.5% of cases of early cancer examined. However, the CEA level in the gastric juice was significantly higher in patients with gastric cancer than in normal subjects or patients with benign gastric diseases: increased CEA concentrations and outputs in the gastric juice were observed in 54.5% and 40.1%, respectively, of the patients with early gastric cancer, and in 52.9% and 50.0%, respectively, of the patients with advanced gastric cancer. The gross type of gastric cancer had no influence on the gastric CEA level, but the CEA level in the gastric juice tended to be high when the tumors were large, or when they had little or no glandular structure or high mucussecreting activity.

Value of Serum Alpha-Fetoprotein and Ferritin in the Diagnosis of Hepatocellular Carcinoma

Alpha-fetoprotein (AFP) and ferritin in the serum were determined by radioimmunoassay and enzyme immunoassay, respectively, in 224 healthy subjects, 55 patients with benign hepatic disease and 44 patients with hepatocellular carcinoma (HCC). The AFP levels in the serum were significantly higher in patients with HCC than in healthy subjects and in patients with benign hepatic disease, but this level was not a satisfactory indicator of small HCC since it was elevated in only 75.0% of the patients with a tumor of less than 3.0 cm in its greatest diameter. Although serum ferritin was elevated in only 56.8% of the patients with HCC, the combination of these two tests raised the diagnostic rate of HCC from 65.9% by serum AFP measurement alone to 88.6% with no appreciable decrease in the specificity for HCC. In particular, it raised the diagnostic rates of lesions of less than 3.0 cm in the greatest diameter from 75.0% by measuring AFP alone to 100%. Thus the combination of AFP and ferritin measurement in the serum is useful for the early detection of HCC.

Immature tumor angiogenesis in high-grade and high-stage renal cell carcinoma.

OBJECTIVES To investigate the correlation between pathologic findings and maturation of the tumor neovasculature of renal cell carcinoma by immunohistochemical studies. METHODS Formalin-fixed and paraffin-embedded specimens from 25 randomly selected patients with renal cell carcinoma were stained with mouse monoclonal antibodies, anti-human CD31, anti-alpha smooth muscle actin (alphaSMA), and anti-human calponin by the indirect immunoperoxidase method. The microvessels were counted in six areas with the higher number of microvessels in each patient at 200x magnification (0.255 mm2 per area). RESULTS The number of CD31-positive microvessels in grade 3 tumors was significantly lower than those in grade 1 or 2 tumors (P = 0.003222 and P = 0.043217, respectively). The CD31-positive microvessel counts of those of higher stage, tumor size greater than 4.5 cm, or non-clear cell type were significantly lower than tumors of lower stage, size less than 4.6 cm, or clear cell type. In the grade 3 tumors, the expression ratio of the number of alphaSMA-positive microvessels to the number of CD31-positive microvessels was significantly decreased compared with grade 1 or 2 tumors (P = 0.000011 and P = 0.000000, respectively). The expression of calponin in the tumor neovasculature was not observed. The expression ratios of the number of alphaSMA-positive microvessels to the number of CD31-positive microvessels in higher stages, larger tumor sizes, or non-clear cell types were significantly decreased. CONCLUSIONS The tumor neovasculature of high-grade and high-stage tumors was immature. These results imply that high-grade tumors of renal cell carcinomas may be susceptible to antiangiogenesis therapy inducing apoptosis of immature tumor vessels.