Abundant expression of immunoreactive endothelin 1 in mammary phyllodes tumor: possible paracrine role of endothelin 1 in the growth of stromal cells in phyllodes tumor.

Immunoreactive endothelin 1 (irET-1) concentrations were measured in extracts prepared from 4 phyllodes tumors and 14 fibroadenomas. irET-1 was detectable in all tissue extracts by specific radioimmunoassay, and the mean concentration of irET-1 was 18-fold and 27-fold higher in tissue extracts from phyllodes tumors than in those from intracanalicular fibroadenomas and pericanalicular fibroadenomas, respectively. Reverse-phase high-performance liquid chromatography coupled with radioimmunoassay in the extracts from phyllodes tumors revealed one major irET-1 component corresponding to human standard ET-1. Furthermore, immunocytochemical staining for ET-1 revealed that numerous ET-1-immunoreactive cells were seen in the epithelial cells but not in the stromal cells, suggesting that ET-1 is synthesized by the epithelial component of phyllodes tumors. A possible paracrine role of ET-1 in the growth of this rare tumor which is characterized by its prominent stromal cellularity is discussed.

Involvement of MMP-7 in invasion of pancreatic cancer cells through activation of the EGFR mediated MEK-ERK signal transduction pathway.

AIMS: To clarify the involvement of matrix metalloproteinase-7 (MMP-7) in cell dissociation and the subsequent invasion of pancreatic cancer cells. METHODS: Western blotting, in vitro invasion assay, immunocytochemistry, and immunohistochemistry were performed in pancreatic cancer cell lines or pancreatic cancer tissue. RESULTS: The active form of the MMP-7 protein was expressed exclusively in the conditioned medium of dissociated (PC-1.0 and AsPC-1) pancreatic cancer cells, whereas proMMP-7 protein was only detected in the conditioned medium of non-dissociated (PC-1 and Capan-2) cells. Both intracellular and conditioned medium localised MMP-7 was greatly reduced by treatment with the epidermal growth factor receptor (EGFR) inhibitor AG1478 and the mitogen activated protein kinase kinase (MEK) inhibitor U0126 in pancreatic cancer cells. MMP-7 treatment significantly induced the disruption of tight junction (TJ) structures and subsequent cell dissociation, and activation of the EGFR mediated MEK- ERK (extracellular signal regulated protein kinase) signalling pathway in the non-dissociated pancreatic cancer cells. Moreover, the strong in vitro invasiveness of dissociated cells was inhibited by AG1478 and U0126 treatment, whereas the weak invasiveness of non-dissociated cells was apparently induced by MMP-7 treatment. In addition, MMP-7 expression was stronger at the invasive front than at the centre of human pancreatic tumours. CONCLUSION: MMP-7 is involved in cell dissociation and the subsequent invasion of pancreatic cancer cells. It induces the disruption of TJ structures and forms a positive feedback loop with activation of the EGFR mediated MEK-ERK signalling pathway.

Light microscopic immunohistochemical analysis of the distribution of group II phospholipase A2 in human digestive organs.

The light microscopic and immunohistochemical distribution of human Group II phospholipase A2 (M-PLA2) in digestive organs of both human fetus and adult, with a new monoclonal antibody (MAb) against M-PLA2, was investigated semiquantitatively. The immunoreactivity was distributed similarly in the adult and fetal epithelium of the esophagus, duodenum, and small intestine, and in the acinar, islet, and duct cells of the pancreas. The epithelium of adult gallbladder was immunoreactive. Paneth cells, especially the secretory apparatus, were strongly immunoreactive. Hepatic Küpffer cells and macrophages of the adult spleen were also immunoreactive. These results suggest that these cells contain secretory-type Group II PLA2, which may be involved in host defensive mechanisms, such as phagocytosis in human digestive organs. In the adult colon, the immunoreactivity was observed only in the ascending colon and was not found in the transverse, descending, sigmoid, or rectal colon. The immunoreactivity was not found in fetal colon. Similarly, immunoreactivity was found in hepatocytes and Küpffer cells of adult liver but not in fetal liver. By contrast, strong immunoreactivity was observed in the epithelium of the fetal stomach but not in adult stomach except in gastric neck cells. This suggests that the expression of M-PLA2 may be related to cell differentiation in particular organs.

Induction of multifocal pancreatic cancer after inoculation of hamster pancreatic cancer cell line (PC-1) into a defined area of homologous pancreatic tissue.

Inoculation of the pancreatic cancer cell line PC-1 into a defined area of the tail of the hamster pancreas led to development of multiple cancerous foci in the body of the pancreas along the main pancreatic duct. Each carcinoma showed identical histological characteristics and was completely separated from the others by regions of normal pancreatic tissue. This observation suggests spread of the cancer from the primary site in discontinuity through the ductal system of the pancreas.

Comparative histopathological findings in the pancreas of cigarette smokers and non-smokers.

Although a correlation has been suggested between cigarette smoking and pancreatic cancer, studies on pathological changes in the pancreas of smokers are fragmentary. In the present study we examined histopathologically 73 pancreases obtained by autopsy from 42 heavy cigarette smokers and 31 non-smoker patients. One invasive adenocarcinoma (2 cm in diameter) and three small carcinomas (2-5 mm in diameter) were found in smokers and one small carcinoma in a non-smoker patient. Although the incidence of pancreatic cancer in smokers was higher than in non-smokers, the difference was statistically not significant. Of smokers with pancreatic cancer, 2 had lung cancer, 1 skin cancer, 1 colon cancer and 1 was free of any malignancies. Ductal changes, including mucinous or squamous cell metaplasia and papillary hyperplasia, were found with equal frequencies in both groups of patients. The type and the incidence of these ductal alterations were not related to smoking but to the age. Our results do not indicate that cigarette smoking increases the incidence of pancreatic cancer, although, the limited number of the sections of the pancreas examined, as well as exclusion of other important variables, such as alcohol, diet and diabetes weaken the value of this study.

Production of group II phospholipase A(2) in advanced gastric cancer.

We examined the immunohistochemical expression of membrane-associated phospholipase A(2) (M-PLA(2)), belonging to group II PLA(2), in 44 advanced gastric cancers, using the ABC method and monoclonal antibody anti-human M-PLA(2). M-PLA(2) mRNA was also examined in the same rumours by Northern blot analysis. In addition, the content of M-PLA(2) protein and prostaglandin E(2) (PGE(2)) in the malignant lesion and in the non-malignant gastric mucosa was examined. The expression was detected in cancer cells in 31 out of 44 advanced gastric cancer tissues (70.4%) by the ABC method. M-PLA(2) mRNA was detected in 36 out of 44 gastric cancer tissues (81.8%), and the density was observed to be higher in tumour tissues than in the adjacent nonmalignant gastric mucosa. The M-PLA(2) protein was detected both in malignant tissues and in non-malignant gastric mucosa, and the content of M-PLA(2) protein was significantly higher in malignant tissues than in the non-malignant gastric mucosa. There was a significant positive correlation between the expression of M-PLA(2) mRNA and the amount of M-PLA(2) protein. PGE, was also detected in the malignant tissues and in the non-malignant mucosa. The content of PGE, was significantly higher in the former. These results indicate that M-PLA(2) is produced both in malignant and nonmalignant cells of the stomach, the former producing higher amounts of this enzyme than the latter. M-PLA(2) may be involved in cancer progression through its function or through the function of products of this enzyme's action such as PGE(2) in gastric cancer.

Prognostic values of MUC-1 molecule expressing cytokine receptor-like epitope and DF3 in patients with gastric carcinoma.

As recently reported, DF3/MUC-1 molecules having cytokine receptor-like sequences (CRL) at the extracellular region, are likely to function in signal transduction pathways. To elucidate the functional significance of CRL expressed on the DF3/MUC1 molecule, immunohistochemical localization of CRL and/or DF3 was investigated in cases of 115 patients with gastric carcinomas, treated by surgical resection. CRL was detected in 65 of 115 patients (56.5%), DF3 in 85 (73.9%), and both DF3 and CRL in 52 (45.2%). The combined immunohistochemical analysis of CRL and/or DF3, revealed that simultaneous expression of DF3 and CRL (DF3+/CRL+) significantly correlated to lymph node metastasis and to blood vessel invasion, and that patients with DF3+/CRL+-tumors survived for a significantly shorter period after surgery than did the other three groups (DF3+/CRL-, DF3-/CRL+, and DF3-/CRL-). Multivariate analysis showed independent prognostic significance for DF3+/CRL+ expression (hazard ratio [HR]=2.733, P=0.0085), and surgical cure (HR=4.334, P=0.003). To investigate the biological role of the simultaneous expression of DF3 and CRL, we constructed DF3-/CRL+ (NR-MC-38) and DF3+/CRL+ (R-MC-38) cells by transducing a mouse colon adenocarcinoma cell line MC-38 expressing neither DF3 nor CRL with MUC-1 cDNA containing ten tandem repeats (R-MC-38) or MUC-1 cDNA devoid of tandem repeats (NR-MC-38). R-MC-38 (DF3+/CRL+) cells were more invasive than NR-MC-38 (DF3-/CRL+) and MC-38 (DF3-/CRL-) cells. When these transfectants were incubated with pAb CRL, the invasiveness of R-MC-38 (DF3+/CRL+) was strikingly elevated over the case with native MC-38 (DF3-/CRL-) and NR-MC-38 (DF3-/CRL+) cells. The pAb CRL-induced invasiveness of R-MC-38 cells was inhibited by adding mAb DF3 or CRL peptides together with pAb CRL. These results suggest that an expression of DF3/MUC1 is highly associated with cell-invasiveness, and the DF3/MUC1-associated invasiveness is amplified by CRL. Thus DF3+/CRL+-MUC-1 molecule seems to be closely involved in a poor prognosis for gastric cancer patients.

Presence of RON receptor tyrosine kinase and its splicing variant in malignant and non-malignant human colonic mucosa.

The presence of RON and its variant isoform in malignant and non-malignant human colonic tissues was examined by immunohistochemistry using paraffin-embedded sections and RT-PCR analysis followed by direct sequencing of PCR product using RNAs isolated from frozen tissues. In normal colonic mucosa, RON was uniformly expressed in crypt cells, especially in the bottom of crypta. On the other hand, the expression was distributed heterogeneously in adenomas and in colon cancer. The expression of RON was significantly related to the degree of differentiation of colon cancer and the deletion of the expression was observed in colon cancer specimens with high incidence. The RT-PCR analysis of RNA isolated from non-malignant and malignant colonic tissue revealed the presence of two RON mRNA isoforms with 432-bp and 286-bp. Direct sequencing of major product of 432-bp was revealed to be identical to that of human wild-type RON. On the other hand, major product of 286-bp was revealed to be almost identical to that of a splicing variant of RON transcript which has been found in human gastric cancer cell line, KATO-III. The results obtained in this study may indicate that both wild-type RON and its variant isoform play an important role in regulating the normal function of colonic mucosa such as differentiation and motile activity and the expression of both wild-type RON and its variant isoform could be considered to be reduced during malignancy of human colonic mucosa.

Comparative studies on the expression of gastrointestinal-cancer-associated antigen, PA8-15, CA19-9 and the blood-group antigens in non-malignant and malignant human pancreatic tissues.

The expression of PA8-15 antigen and the blood-group-related antigens A, B, O, Le(a), Le(b), Le(x), and Le(y), as well as CA19-9, were examined in the normal pancreas and in specimens from benign and malignant pancreatic tissue by the avidin-biotin-immunoperoxidase technique. A correlation was found between the expression of PA8-15, Le(a), and CA19-9 in some cases. However, in the cancer tissues in which neither Le(a) nor CA19-9 could be demonstrated, strong expression of PA8-15 was observed. The reactivity of monoclonal antibody (mAb) PA8-15 with pancreatic cancer tissue was not inhibited by the preincubation of the sections with the mAb anti-Le(a) (CO514) and mAb CA19-9 (CO19-9) indicating that the epitope recognized by PA8-15 is different from that detected by the other two antibodies. Moreover, unlike Le(a) and CA19-9, PA8-15 was also expressed in cancer cells of patients of the Le(a-b-) type. The results suggest that mAb PA8-15 recognizes a sialylated molecule related to Le(a) but different from CA19-9, and seems to be an additional useful marker for pancreatic cancer.

Expression of blood group-related antigens, ABH, Lewis(a), Lewis(b), Lewis(x), Lewis(y), CA19-9, and CSLEX1 in early cancer, intestinal metaplasia, and uninvolved mucosa of the stomach.

The expression of blood group-related antigens, A, B, H type 2, Lewis type 1 (Lewis(a) [Le(a)] and Lewis(b) [Le(b)]), Lewis type 2 (Lewis(x) [Le(x)] and Lewis(y) [Le(y)]), sialylated Le(a) (CA19-9), and sialylated Le(x) (CSLEX1), was analyzed sequentially with immunohistochemical methods in early gastric cancer, intestinal metaplasia, and uninvolved gastric mucosa obtained from 35 surgical specimens of patients who underwent gastrectomy. The high incidence of the inappropriate expression of Lewis type 1 antigens and the deletion of H and Lewis type 2 antigens was observed similarly in patients with cancer and intestinal metaplasia. The acquisition of CA19-9 and CSLEX1 and the deletion of B antigen frequently were found in intestinal-type cancer and all types of intestinal metaplasia. The simultaneous deletion of A antigen was detected only in the combination of intestinal-type cancer and incomplete-type intestinal metaplasia. Thus the present study shows that similar changes of tissue antigenicities exist in early gastric cancer and intestinal metaplasia.

Changes in production of interleukin-1 and interleukin-2 associated with obstructive jaundice and biliary drainage in patients with gastrointestinal cancer.

Increased susceptibility to infection in patients with obstructive jaundice has been well documented in vitro and in vivo. Nevertheless, an underlying mechanism for immunocompromise in these patients has not been identified. This study was undertaken to evaluate the production of two important immunoregulatory molecules, interleukin-1 (IL-1) and interleukin-2 (IL-2), by peripheral blood mononuclear cells in cancer patients with obstructive jaundice before and after percutaneous transhepatic biliary drainage (PTBD). After decompression with PTBD, IL-1 and IL-2 production was significantly increased (IL-1: from 7.9 +/- 4.9 to 13.9 +/- 4.9 U/ml, p less than 0.05; IL-2: from 8.8 +/- 4.9 to 14.1 +/- 6.5 U/ml, p less than 0.05). There was a positive correlation between IL-1 and IL-2 production (r = 0.424, p less than 0.05). The production of both interleukins correlated negatively with serum total bilirubin level (IL-1 r = -0.478, p less than 0.05; IL-2: r = -0.482, p less than 0.05) and positively with high-density lipoprotein cholesterol in serum (IL-1: r = 0.505, p less than 0.01; IL-2: r = 0.494, p less than 0.05). IL-2 production also correlated positively with serum albumin levels (r = 0.511, p less than 0.01). These results suggest that hyperbilirubinemia and abnormal lipid metabolism may be associated with impaired interleukin production, which may result in an increased susceptibility to infection during obstructive jaundice.

No-touch isolation technique reduces intraoperative shedding of tumor cells into the portal vein during resection of colorectal cancer.

BACKGROUND: The mutant-allele-specific amplification (MASA) method is capable of detecting 1 genetically altered tumor cell among thousands of normal cells. The MASA enabled us to detect occult tumor cells undetectable by histopathologic examination of lymph nodes and blood samples. METHODS: To investigate whether tumor manipulation during operation enhances cancer cell dissemination into the portal vein with use of MASA and to assess the effect of the no-touch isolation technique in the treatment of colorectal cancers, 27 colorectal cancers (17 were operated on conventionally and 10 were operated on according to the no-touch isolation technique) were screened for mutations in K-ras or p53. We next examined blood samples of the portal vein collected before, during, and after manipulation of tumors, using MASA to look for the specific mutation found in the primary tumors. RESULTS: Somatic mutations were identified in 18 of these primary tumors (11 were in the conventional resection technique group and 7 were in the no-touch isolation technique group). In 8 of 11 (73%) conventional resection technique cases, we identified the same genetic alteration of the primary tumor in the portal blood during operation, whereas only 1 patient (14%) in the no-touch isolation technique group had a positive result. CONCLUSIONS: The no-touch isolation technique may be useful to prevent cancer cells from being shed into the portal vein during surgical manipulation.

Relationship between plasma cytokine concentration and multiple organ failure in patients with acute pancreatitis.

The dynamic aspects of circulating cytokines and cytokine modulators and their relationship with development of multiple organ failure (MOF) in patients with acute pancreatitis were analyzed. All cytokine and C-reactive protein levels in the circulation were higher than those in the MOF group. In particular, plasma concentrations of soluble tumor necrosis factor receptors (sTNF-RI and sTNF-RII) were significantly higher in patients with MOF than in those without even at admission. Furthermore, plasma concentrations of sTNF-Rs and interleukin-1 (IL-1) receptor antagonist (IL-1ra) were much higher than those of their counterparts, TNFalpha and IL-beta, respectively. These results suggest that the plasma concentrations of sTNF-Rs are useful predictors for the development of MOF, and actions of TNF-alpha and IL-1beta could be regulated by their modulators (soluble receptor and receptor antagonist, respectively) in the pathologic condition of severe acute pancreatitis.

Supernumerary lactate dehydrogenase isozymes in human gliomas.

Supernumerary bands in agarose gel electrophoresis of lactate dehydrogenase (LDH) were frequently observed in extracts of human gliomas. The supernumerary fractions which migrated cathodic to LDH-2 and/or cathodic to LDH-3 were designated LDH-2' and LDH-3'. These extra bands were clearly seen in certain gliomas and less distinctly in others, being more frequent in primitive or undifferentiated tumours. The extra bands seen in gliomas differ from the LDH-X of the testis, but LDH-2' seemed correspond to LDH-Z in placenta, hydatidiform mole, and choriocarcinoma. These sub-bands are interpreted as being produced by gliomas and as oncofetal enzymes.

Effects of acute and repeated alcohol ingestion on hypothalamic-pituitary-gonadal and hypothalamic-pituitary-adrenal functioning in normal males.

We investigated the effects of acute and repeated alcohol ingestion on plasma levels of hormones associated with the functioning of the hypothalamic-pituitary-gonadal (HPG) and hypothalamic-pituitary-adrenal (HPA) systems in normal males. In the first experiment, 7 normal male subjects were given ethanol (1.3 g/kg) in the form of a 43% alcohol solution of whiskey and water over a 30-min period (from 19:00 h to 19:30 h); blood samples were collected 30 min and immediately before the beginning of alcohol ingestion and then at intervals of 30 min for 180 min. Blood ethanol levels rose sharply and reached their maximum at 60 min, remaining above 1.0 mg/ml until 180 min. Prolactin levels increased, reaching a peak at 60 min, gradually returning to the initial value at 180 min. Decreased testosterone levels were observed only at 30 min. Luteinizing hormone (LH), adrenocorticotrophic hormone (ACTH) and cortisol levels did not show any increases. In the second experiment, 9 normal males were given the same dose of alcohol, but this was given on 7 consecutive evenings and the hormonal changes were examined on the 1st and 7th days, only at 30 and 60 min after alcohol ingestion began (during the period that blood ethanol levels were ascending to their peak). The results on the 1st day reconfirmed the findings in the first experiment and on the 7th day, the last alcohol ingestion produced increases in prolactin levels and decreases in testosterone levels at 30 and 60 min, but did not change other hormone levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Vitamin B12 accelerates re-entrainment of activity rhythms in rats.

The effects of methyl vitamin B12 (5-6 mg/kg, p.o.) on the entrainment of circadian running wheel activity rhythm to a new lighting schedule were measured in rats. After the light-dark (LD) cycle was abruptly reversed, rats given vitamin B12 took less time to entrain their circadian locomotor activity rhythm to the new cycle than did controls. This result indicates that vitamin B12 accelerates the reentrainment of the mammalian circadian activity rhythm following an abrupt change in the environmental LD cycle.

Neuronal and glial proteins in medulloblastomas. II. Heterotransplantation of human medulloblastoma.

Seven resected medulloblastomas were inoculated into hereditary athymic (nude) and asplenic athymic (Lasat) mice. From these, only one transplantable tumor line was obtained, in nude mice. We studied the transplanted medulloblastoma immunohistochemically and by electron microscope. In addition, using enzyme immunoassay, the gamma-enolase contents of the transplanted medulloblastomas were measured and the results were compared with those of transplanted glioblastomas. Neurofilament proteins (NF68Kd, NF160Kd) and gamma-enolase positive cells were maintained in the successful transplants of medulloblastomas, but the GFAP positive cells were lost. Under the electron microscope, a few cells with ten nanometer intermediate filaments and microtubules were observed in the transplanted tumor. The medulloblastoma possesses some of the neuronal characteristics which are maintained in the transplants in nude mice.

Association of plasma free-3-methoxy-4-hydroxyphenyl (ethylene)glycol, natural killer cell activity and delirium in postoperative patients.

We measured and compared levels of plasma free 3-methoxy-4-hydroxyphenyl (ethylene)glycol (pMHPG), a major metabolite of noradrenaline, and natural killer (NK) cell activity in 26 patients prior to their undergoing an operation for cardiovascular diseases; 11 of whom expressed delirium and 15 who did not. In conclusion, we found that pMHPG levels before an operation were higher in patients with postoperative delirium than in the patients without, while NK cell activity showed no difference between the two groups. It is possible that hyperactivity of noradrenargic neurons is connected with the development of postoperative delirium. Furthermore, we considered that measurement of pMHPG level before operation might be a useful tool to predict the occurrence of postoperative delirium.

Evaluation of serum CA125 values in healthy individuals and pregnant women.

CA125 is an antigenic determinant associated with human epithelial ovarian carcinoma. This study was undertaken to evaluate the distribution of serum CA125 levels and the effect of smoking on these levels among healthy individuals and clarify the relation of maternal serum CA125 level and pregnancy. Among 552 healthy individuals, the distribution of serum CA125 values was demonstrated to resemble logarithmic normal distribution. Analysis of variance about age and sex revealed apparent elevation of values for women under 49 years of age in comparison with those for women over 50 years of age and men. Values for these two groups were 143 units/ml for the former and 32 units/ml for the latter, with a 99.7% confidence limit. These values exclude 99.3% of the former and 99.7% of the latter. Serum CA125 values were not affected by smoking. The measurement of serum CA125 levels in 71 pregnant women disclosed a significant elevation during the first trimester in comparison with nonpregnant women under 49 years of age. These results indicate that CA125 values must be deliberatively evaluated in young women, especially during first trimester of pregnancy.