Kuniyasu Fukuzawa

Expression of nitric oxide synthase in gastric cancer

The expression of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) in samples of normal gastric mucosa and gastric cancer were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and semi-quantitative Western blot. In normal gastric mucosa, eNOS protein was found in all samples examined (mean, 70.2 +/- 60.1), relative to a standard protein. In gastric cancer specimens, eNOS protein was also detected in all samples, but the quantity (86.5 +/- 76.6) was not different from that found in samples of normal mucosa. The quantity of eNOS in gastric cancer tissues was negatively correlated with serosal invasion. iNbS mRNA, detected in nine of 18 cases, was slightly related to massive lymph node metastasis (n1-3 vs. n4). Neither tumor necrosis factor alpha (TNF-alpha) mRNA nor interleukin-6 (IL-6) mRNA was related to the expression of iNOS mRNA. These results suggest that iNOS not eNOS plays a role in gastric cancer tumor extension, but iNOS mRNA appears not to be induced by either TNF-alpha or IL-6.

Expression of facilitative glucose transporter 1 mRNA in colon cancer was not regulated by k-ras

The expression of facilitative glucose transporter isoforms in colon adenocarcinoma and the possible role of k-ras in inducing GLUT (glucose transporter) mRNA were studied. RT-PCR demonstrated GLUT2 and GLUT3 expression in 100% of the ten normal colon mucosa samples but detected no GLUT1 mRNA. By contrast, GLUT1 mRNA was detected in all 20 (100%) colon cancer samples examined. GLUT4 mRNA was not detected in either normal mucosa or colon cancer tissues. Semiquantitative PCR demonstrated equal amounts of GLUT2 and GLUT3 mRNA in both normal mucosa and colon cancer samples. A point mutation in codon 12 of k-ras was detected in only six of the 20 (30%) colon cancer samples. Thus, a major difference between normal colon epithelia and colon cancer was the acquisition of GLUT1 expression, which was unlikely to have been induced by a point mutation in codon 12 of k-ras.

High incidence of synchronous cancer of the oral cavity and the upper gastrointestinal tract

A high incidence of synchronous esophageal or gastric carcinoma in preoperative patients with carcinoma of the oral cavity was reported. Esophageal carcinoma was found in seven out of 56 patients (12.5%) and gastric cancer in five patients (8.9%) by videoendoscopy aided with lugol staining in the esophagus and indigocarmine solution in the stomach, although all patients were completely asymptomatic for these lesions. All patients were male, regular drinkers and heavy smokers. The depth of invasion of such tumors was limited to either mucosa or submucosa. Those esophageal and gastric lesions beside the primary oral cancers were positive for p53 protein by immunohistochemistry. Careful preoperative evaluation of not only the esophagus but also the stomach should be a routine procedure in patients with carcinoma of the oral cavity.

Long-term effect of radical gastrectomy on nutrition and immunity

The long-term effects of gastrectomy on the nutritional and immunologic status were prospectively studied in 79 gastric cancer patients who underwent curative gastrectomy and were followed by us after operation for an average of 5 years and 3 months. The percent of actual weight to pre-illness normal weight was lower than 95% in 80% of all study patients. Retinol binding protein, prealbumin, and albumin were lower than normal in 17%, 26%, and 26% of the patients, respectively. The mean values of the percent normal weight, retinol binding protein, and prealbumin were significantly lower in the totally gastrectomized patients than in the subtotally gastrectomized ones (P<0.01). The procedures of reconstruction did not affect the nutritional status except for the prealbumin level which was significantly decreased in Roux-en-Y cases than in interposed cases of totally gastrectomized patients. Cell-mediated immunological alterations after gastrectomy were observed in 31%, 37%, and 71% of all patients for OKT3 subpopulation, OKT4/OKT8 ratio, and blastogenesis by phytohemagglutinin, respectively. A multivariate analysis revealed that the long-term immunity of the gastrectomized patients after operation was not affected by the levels of albumin and rapid turnover proteins but by the splenectomy and weight loss they underwent.

The possible role of TNF-α and IL-2 in inducing tumor-associated metabolic alterations

This study was conducted to investigate the role of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) in inducing cancer cachexia, and the results were compared with those obtained from our previous study on Fisher 344 rats with methylcholanthrene-induced sarcoma. Three groups of male Fisher 344 rats received one of the following regimens: 4×104 IU of human recombinant TNF-α per rat per day subcutaneously (sc) for 5 consecutive days (n=5), 3.5×105 U human recombinant IL-2 per rat per day sc for 14 consecutive days (n=5), or normal saline (n=5). The activities of both phosphoenolpyruvate carboxykinase (PEPCK) and malic enzyme (ME) were increased slightly in the IL-2 group. Furthermore, LPL activity was significantly increased in the adipose tissue of the TNF group and in the cardiac muscle of the IL-2 group, but not in that of the TNF group. These results show that there is a significant difference between the metabolic alterations seen in the tumor-bearing state and those induced by either TNF-α or IL-2 alone. Thus, it is unlikely that IL-2 or TNF-α is the sole mediator of cancer cachexia in this tumor and rat model.

Alterations in DNA proliferation in gastric stump mucosa with special reference to topography

BACKGROUND Gastric stump carcinoma occurs more frequently at the anastomotic site than at other areas of the gastric remnant. This study was conducted to determine whether alterations in cell dynamics could be detected in the normal-looking gastric remnant mucosa and to ascertain any difference in cell dynamics between the anastomotic site and other area. METHODS Sixty-three patients who underwent curative subtotal gastrectomy for gastric adenocarcinoma were examined 8 to 130 months after operation. Mucosa at the anastomotic site and at the greater curvature were endoscopically biopsied. Cell dynamic changes were examined by flow cytometry, bromodeoxyuridine in vitro labeling, and oncogene (K-ras and c-myc) amplification and translocation by Southern blot analyses. RESULTS Inflammatory changes were observed in more than 60% of the cases both at the anastomotic site and at the greater curvature both macroscopically and microscopically. Blood flow, measured by laser Doppler flowmeter, was significantly decreased at the anastomotic site compared with the greater curvature (25.0 +/- 8.1 and 32.2 +/- 9.4 mg/min/100 mg tissue, respectively; p < 0.05). Analysis of DNA histograms obtained by flow cytometry revealed a small peak at the right shoulder of 2N peak in 33% of the specimens obtained from the anastomosis site and in 15% of those from the greater curvature (p < 0.05). The number of cells at S phase measured by bromodeoxyuridine in vitro labeling was also significantly greater at the anastomotic site than at the greater curvature. None of the five patients with aneuploid DNA histograms at the anastomotic site had detectable amplification or translocation of K-ras or c-myc gene in mucosal tissue. CONCLUSIONS More marked changes in the mucosal cell dynamics observed at the anastomotic site may partially explain the higher incidence of gastric stump carcinoma at the anastomotic site.

Long-term total parenteral nutrition and osteoporosis : report of a case

A patient who had been supported with total parenteral nutrition (TPN) for over 8 years is herein presented, with emphasis on the changes observed in calcium metabolism. The patient was a 31-year-old female, who had undergone a subtotal jejunal and ileal resection for superior mesenteric artery occlusion. TPN was started soon after the surgery. She had been on TPN support for 105 months. Back pain developed at 97 months after the initiation of TPN. During her course, the serum calcium levels were judged to be within the normal ranges, while the 1α, 25(OH)2Vit.D declined. Intermittent hypercalciuria was occasionally observed. Both the serum level of calcium and urinary calcium loss correlated closely to the amount of calcium infused, but they were not influenced by the amount of vitamin D (ergocalciferol) received. The serum level of parathormone and calcitonin were also within the normal ranges. The patients vertebral bone, which was obtained at autopsy, revealed histopathological changes characteristic of osteoporosis. Based on the above, we conclude that a careful monitoring of the amount of calcium infused is called for to prevent bone disease in patients on long-term parenteral nutrition.

Role of insulin resistance in decreasing lipoprotein lipase activity in tumor-bearing rats

The role of insulin resistance in the tumor-induced decrease in tissue lipoprotein lipase (LPL) activity was studied in vivo and vitro in methylcholanthrene-induced sarcoma-bearing rats. Intraperitoneal (ip) injection of 2 U of regular insulin resulted in high-adipose LPL activity in control rats (CTR) of 122.0±42.4 U/mg tissue, but it had little effect on tumor-bearing rats (TBR), which showed a value of only 9.6±5.5 U/mg tissue (P=0.002). When adjusted for serum insulin concentrations, adipose LPL activity remained significantly different between the TBR and CTR at 0.19±0.17 and 0.78±0.29 U/mg tissue, respectively (P=0.02). Following the in vitro incubation with either 1.44 g/l glucose of 1×10−8 U insulin of adipose tissue fragments obtained from the TBR and the CTR, measurable LPL activity was maintained in the tissue from the CTR for 2h but not in that from the TBR. These results suggest that the decreased LPL activities seen in the tumor-bearing state may be mediated by insulin resistance.

The effects of a biological response modifier, OK-432, on tumor-induced alterations in the host metabolism

The effects of multicytokine inducer, OK-432, on tumor-induced metabolic alterations were studied by assessing three key regulatory enzymes of gluconeogenesis, de novo fatty acid synthesis and the triglyceride clearance pathways. Two Klinish Einheit (KE) of OK-432 was subcutaneously injected on alternate days, for 10 days, into Fischer 344 rats with or without methylcholanthrene-induced sarcoma. At the time of sacrifice, the tumors accounted for approximately 23% of their total body weight. The injections of OK-432 did not affect the amount of food intake in either the tumor bearers or the controls. The tissue lipoprotein lipase activities in the epididymal fat pads of the tumor bearers were significantly decreased compared with the controls (P<0.01). Phosphoenolpyruvate carboxykinase activity in the liver was significantly increased (P<0.01), while malic enzyme activity tended to be decreased in the tumor bearers compared with the controls. However, there were no significant differences in those activities depending on the OK-432 injections, even though OK-432 induced tumor necrosis factor (TNF) and increased cytotoxic activities in the mesenteric lymph nodes as well as in the spleen. Thus, although the role of monokines in inducing cancer cachexia is not yet clearly understood, OK-432 was not able to revert the tumor-induced metabolic alterations which lead to tissue wasting and cancer cachexia.

Laminin Levels and Stage of Vascular Invasion as Predictive Factors of Liver Metastasis in Colon Cancer

The glycoprotein laminin mediates the adhesion of tumor cells to the basement membrane prior to invasion. In this study, we determined the laminin levels of the peripheral and returned venous blood in 46 colon cancer patients without detectable liver metastasis. The laminin levels of both peripheral and returned blood were significantly higher in the cases with vascular invasion compared to the group without it. However, no correlation was observed between the stage of vascular invasion and laminin levels.