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We examined secretion, mRNA expression, and histologic localization of interleukin-8 (IL-*) and growth-related gene product-alpha (GRO alpha) in the Helicobacter pylori-infected gastric antral mucosa. Antral biopsies were obtained from an area of endoscopically intact mucosa. Significantly higher levels of IL-8 and GRO alpha were secreted in organ cultures from patients with H. pylori infection, and their elevation was prominent in patients with duodenal ulcer. There was a significant association between these alpha-chemokine levels and histologic grades of activity, inflammation, and H. pylori density. In fresh antral biopsies, IL-8 and GRO alpha mRNA expression was detected more frequently in H. pylori-infected patients compared with those without infection. Immunofluorescence microscopy showed localization of IL-8 and GRO alpha proteins in gastric epithelial cells and infiltrating CD68+ macrophages. In the chemotaxis assay, a significant positive correlation was found between neutrophil migration induced by the organ culture supernatants and their contents of IL-8 and GRO alpha. After H. pylori eradication, a significant decrease was observed in IL-8 and GRO alpha levels detected in organ cultures. In conclusion, mucosal alpha-chemokine activity correlates well with histologic severity of H. pylori-associated antral gastritis and can be used to predict the effects of H. pylori eradication therapy.

The intercalated discs of working myocardium and Purkinje fibers of the monkey heart were examined by scanning and transmission electron microscopy. The NaOH/ultrasonication technique resulted in the digestion of connective tissue and a separation of the intercellular junctions of intercalated discs, such that these could be visualized three-dimensionally. The intercalated discs of ventricular myocytes, atrial myocytes and Purkinje fibers vary considerably in number and configuration, as do the intercalated discs of the three different layers of the ventricular myocardium. Myocytes in the subepicardial, middle and subendocardial layers of the ventricle have 1-3, 4-5 and 5-6 intercalated discs at the end of these cells, respectively. Those in the endocardial layer are characterized by the presence of small laterally-placed intercalated discs. Atrial myocytes and Purkinje fibers usually only have 1-2 intercalated discs. Individual intercalated discs in ventricular myocytes have complicated stairs with 10-30 steps and corresponding risers, while those of atrial myocytes and Purkinje fibers have simple stairs with 1-3 steps and risers. Steps equivalent to the plicate segments are characterized by densely-packed microplicae and finger-like microprojections which greatly increase surface area in ventricular myocytes. Microprojections in atrial myocytes and Purkinje fibers are sparse by comparison. Risers equivalent to the interplicate segments containing large gap junctional areas are most numerous in left ventricular myocytes, followed by right ventricular myocytes, Purkinje fibers and atrial myocytes in decreasing order. The geometric arrangement of the various types of myocytes may be related with impulse propagation. Large intercalated discs of cell trunks and series branches may participate in longitudinal propagation, while small laterally-placed ones may be the site of transverse propagation.

Lymphokine-activated killer (LAK) cell activity and messenger RNA (mRNA) expression for interleukin-2 (IL-2) were analyzed using lamina propria mononuclear cells (LPMC) in inflammatory bowel disease patients. Compared with control LPMC, Crohn's disease (CD) LPMC exhibited significantly higher levels of LAK cell activity, whereas ulcerative colitis (UC) cells showed significantly lower levels of cytolytic activity with a difference in the frequency of CD3+, CD56+ and CD3+, CD56+ LAK precursor cells. After incubation with IL-2, the proportion of CD3+, CD56+ lymphocytes continued to be higher in CD cultures and substantially lower in UC cultures. Freshly isolated CD LPMC exhibited significantly higher levels of IL-2 mRNA than controls. However, no significant difference was observed between UC and control cells. The level of IL-2 expression or responsiveness to IL-2 may be responsible for different mucosal immune reactivity between CD and UC patients.

Analysis of cytokine production and function is an indispensable tool to study Crohn's disease and ulcerative colitis. This analysis has generated a tremendous amount of data, but a clear interpretation of results has been hampered by limited attention paid to several patient- and sample-related pitfalls. These include: clinical parameters; the relative value of circulating vs. intestinal cytokine levels; the selection of tissue specimens and their processing method; the specific cellular source of each cytokine; the effect of cytokine inducers; and the technique utilized to obtain qualitative or quantitative data on cytokine protein or mRNA. Once all of these crucial variables are taken into proper perspective, a better understanding will emerge of the true role of cytokines in the pathogenesis of inflammatory bowel disease.

Local interleukin-6 (IL-6) activity was studied using colonic mucosal tissues in inflammatory bowel disease (IBD) and inflammatory control patients. Active IBD specimens exhibited significantly higher IL-6 activity than control specimens in both cultures of isolated lamina propria mononuclear cells (LPMC) and mucosal tissues with an increased number of IL-6-producing cells. However, the activity in inactive IBD or inflammatory controls did not differ from controls. Northern blot analysis demonstrated IL-6 messenger RNA in LPMC and colonic epithelial cells isolated from active IBD specimens but not in control cells. Furthermore, immunofluorescent microscopic study of active IBD specimens showed more conspicuous staining of IL-6 in infiltrating LPMC (mostly CD68+ cells) and colonic epithelial cells. These results suggest that elevation of local IL-6 activity may be a characteristic feature of active IBD and both macrophages and colonic epithelial cells are the major cell types responsible for this phenomenon.

Benzyl 6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside (beta-primeveroside) (1), 2-phenylethyl beta-primeveroside (2), and 2-phenylethyl 6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranoside (beta-rutinoside) (3) were isolated as aroma precursors of benzyl alcohol and 2-phenylethanol from flower buds of Jasminum sambac Ait. The isolation was guided by an enzymatic hydrolysis and GC and GC-MS analyses.

Forty-eight patients with unresectable gastric cancer were enrolled for a randomized trial of FP (n = 24) vs FEP (n = 24) combination chemotherapy with respect to their effects on survival period. Of these, excluding 7 patients, 21 in FP group were treated with 400 mg/m2 UFT po daily and 50 mg/m2 CDDP i.v. on days 1 and 8 every 4 weeks, and 20 in FEP group were treated with 400 mg/m2 UFT po daily, 50 mg/m2 etoposide iv and 50 mg/m2 CDDP i.v. on days 1 and 8 every 4 weeks. In FP group, 10 PR in primary lesions were observed with an overall response rate of 52.4%. In FEP group, 5 PR in primary lesions were observed with an overall response rate of 35.0%. Toxicities over Grade 3 were anorexia (35%), vomiting (25%), and leukopenia (12%), which were manageable. The 50% survival time in FP and FEP group was 233 and 176 days, respectively. Because of the high response rate and prolongation of the survival period, combination chemotherapy with FP seemed to be useful for the treatment of patients with advanced gastric cancer.

Bipolal [1], a new compound with marine antifouling potential, was isolated from the culture broth of Bipolaris sp. The structure was determined to be an eremophilane derivative on the basis of 2D nmr spectroscopy and chemical transformations.

Intestinal T-cell lines were generated from lamina propria mononuclear cells isolated from colonoscopic biopsies in ulcerative colitis patients and controls. In both ulcerative colitis and controls, expanded cells were constituted largely by T-cell receptor alpha beta+, CD4+, CD45RA- (helper), and CD8+, CD11b- (cytotoxic) phenotypes. T-cell receptor V beta gene usage was not significantly changed after cell expansion and no difference was observed between ulcerative colitis and controls. Ulcerative colitis cells, especially those derived from the patients with long-standing disease, showed significantly higher levels of cytotoxicity against the target cells, including those of colonic epithelial origin, and enhanced production of tumor necrosis factor-alpha and interferon-gamma after short incubation with anti-CD3 antibody. Generation of T-cell lines from colonoscopic biopsy specimens may be useful for detailed functional characterization of locally infiltrating T cells in ulcerative colitis patients.

Rat pancreatic islets were allografted in the liver and were studied morphologically in order to evaluate possible reinnervation. Islets isolated from rat pancreas were allotransplanted in the liver of streptozocin-induced diabetic rats via the portal vein. Electron microscopy revealed nerve endings with synaptic vesicles in the transplanted islets 100 days after transplantation, whereas axons in the islets appeared to degenerate several hours after isolation and prior to transplantation. These findings suggest that the nerve endings observed in the transplanted islets regenerate from nerves that innervate the recipient's liver. The tissue specimens were also investigated immunohistochemically using antityrosine hydroxylase antibody, and histochemically by the modified Karnovsky and Roots' method for visualizing acetylcholinesterase. Some nerve endings in the transplanted islets reacted positively to antityrosine hydroxylase antibody. Acetylcholinesterase was visualized in other nerves. These results indicate that norepinephrine- and acetylcholinesterase-containing nerves may reinnervate the transplanted islets.

(R)-Linalyl 6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside and bornyl 6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside were isolated as aroma precursors of linalool and borneol from flower buds of Gardenia jasminoides, guided by enzymatic hydrolysis followed by GC and GC-MS analyses.

New glycosidic aroma precursors (1 and 2) of the main volatile constituents, trans- and cis-linalool 3,6-oxides (linalool oxides I and II), were isolated from oolong tea leaves (Camellia sinensis var. sinensis cv. Maoxie). The isolation was guided by an enzymatic hydrolysis with acetone powder prepared from fresh tea leaves (cv. Yabukita) followed by GC or GC-MS analyses. Chromatographic purification of hot water extracts of the tea leaves on active charcoal, Amberlite XAD-2, and Sephadex LH-20 columns as well as HPLC gave two new glycosides, trans- and cis-linalool 3,6-oxide 6-O-beta-D-xylopyranosyl-beta-D-glucopyranosides (1 and 2).

Three glycosides, 6-O-beta-D-xylopyranosyl-beta-D-glucopyranosides (beta-primeverosides) of the aroma constituents, linalool, 2-phenylethanol, and benzyl alcohol, were isolated as aroma precursors from the tea leaves (Camellia sinensis var. sinensis cv. Shuixian and Maoxie, cultivars for oolong tea). The isolation was guided by acid or enzymatic hydrolysis, and subsequent GC and GC-MS analyses. The linalyl glycoside is the first example of naturally occurring (S)-linalyl beta-primeveroside.

A modified NaOH maceration method for removing extracellular ground substance was applied to scanning electron microscopic (SEM) studies of the mammalian aorta and elastic cartilage. Fixed tissues were freeze-fractured in liquid nitrogen and then immersed in 2N NaOH solution for 4-6 hr at room temperature. Since this method selectively digested proteoglycans, cellular and fibrous elements were observable clearly under the SEM. The elastic laminae of smooth muscle cells of the aortic tunica media were concealed by fine collagen fibrils, while chondrocytes in the elastic cartilage were encapsulated by a dense network of fine collagen fibrils and branching elastin fibers.

A new geranyl glycoside, geranyl 5-O-beta-D-xylopyranosyl-beta-D-glucopyranoside was isolated as an aroma precursor from tea leaves (Camellia sinensis var. sinensis cv Shuixian) for oolong tea. The isolation was guided by a two-phase acid hydrolysis and/or an enzymatic hydrolysis followed by GC and GC-MS analyses.

Flower fragrance compounds were found to be produced from the precursor solution obtained from flower buds by crude enzyme prepared from the flowers at the opening stage. GC and GC-MS analyses showed the formation of volatile aroma constituents from the precursor solution of Jasminum polyanthum F, Jasminum sambac Ait, and Gardenia jasminoides E, but none in the case of Osmanthus fragrans L. The aroma-producing enzyme activity of G. jasminoides rapidly increased to reach the maximum at flower opening stage (stage 4) and decreased within 24 h after flower opening (stage 5). Fragrance precursors of G. jasminoides were suggested not to be mainly β-glucosides of linalool, eugenol, borneol, and isoeugenol based on the results after β-glucosidase and naringinase treatment of the precursor solution. The activity of hydrolytic enzyme(s) such as glycosidase was found to elevate during flower opening to result in the aroma formation.

Phenotypic and functional analysis was performed with lamina propria mononuclear cells (LPMCs) isolated from colonoscopic biopsies in 27 patients with ulcerative colitis (UC). The proportion of T lymphocytes displaying HLA-DR antigens, interleukin 2 (IL-2) receptor, and transferrin receptor was greater in active UC than in control diseases. When LPMCs were cultured with IL-2 or phytohemagglutinin for 72 h, there were no significant differences in the proportion of cells bearing these activation markers between active UC and controls. The proportion of CD56+ cells and lymphokine-activated killer (LAK) cell activity was lower in LPMCs from active UC than in control cells, and depletion of CD56+ cells from control lamina propria cells essentially eliminated LAK cell activity. Mucosal T lymphocytes may be activated in vivo during active inflammation in UC, and lower levels of intestinal LAK cell activity may be related to the decrease of CD56+ cells under these conditions.

The difference between anti poly (ADP-ribose) antibodies was studied in patients with systemic lupus erythematosus (SLE), progressive systemic sclerosis (PSS) and drug-induced lupus (DIL). Radioimmunoassay showed that high concentrations of anti poly (ADP-ribose) antibodies (10.3-22.2%) were induced by phenobarbital, phenytoin, valproic acid (anti-epileptic agent) and procainamide (anti-arrhythmic agent). Poly (ADP-ribose) antibodies were separated by hydroxylapatite column chromatography. The average chain length of the polymer consisted of 2.4, 10.8 and 28.2 ADP-ribose units. Anti poly (ADP-ribose) antibodies from patients with SLE and PSS reacted with 2.4, 10.8, and 28.2 ADP-ribose units in RIA, but those from patients with DIL reacted only with 28.2 ADP-ribose units in RIA. The binding specificity of anti poly (ADP-ribose) antibodies from pregnant women was found to be very similar to that of the antibodies from case of DIL. The present results clearly demonstrated that anti poly (ADP-ribose) antibodies found in systemic autoimmune diseases bound not only to poly (ADP-ribose) with an average chain length of more than 20 ADP-ribose units, but also to oligo (ADP-ribose) with an average chain length of about 2 ADP-ribose units. Anti poly (ADP-ribose) antibodies found in DIL cases and pregnant women, however, bound only to poly (ADP-ribose) with an average chain length of more than 20 ADP-ribose units.

Phagostimulant activity of glycerolipids such as digalactosyldiacylglycerol (DGDG), 6-sulfoquinovosyldiacylglycerol (SQDG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), etc., have been examined using the Avicel plate method for three kinds of marine herbivorous gastropods, the abaloneHaliotis discus, the turban shellTurbo cornutus, and the topshellOmphalius pfeifferi. DGDG showed strong activity for all the test animals. SQDG was much less active than the other glycerolipids for abalone. The turban shell and the top shell responded more or less sensitively to all tested glycerolipids at doses of 10-20μg/sample zone.

The interleukin 2 (IL-2) and interferon-gamma (IFN-gamma) activities of regional lymph node mononuclear cells (LNMC) were studied in 22 patients with gastric cancer. Regional LNMC could produce high levels of IL-2 and IFN-gamma independent of the disease extension or the location of the lymph nodes. The activities of peripheral blood mononuclear cells (PBMC) from cancer patients were decreased especially in advanced disease compared to controls. The lymph node CD4+ cells were responsible for the bulk of IL-2 activity, whereas both CD4+ and CD8+ cells were equally capable of producing IFN-gamma. The proportion of T cells in LNMC was similar to that seen in PBMC, however, CD4+ cells predominated over CD8+ cells in the lymph nodes, contributing to higher activities of IL-2 and IFN-gamma in LNMC. Furthermore, significant cytotoxic activity could be induced from PBMC using the culture supernatant derived from autologous LNMC in cancer patients. This study suggests that regional lymph nodes are strategically located near the tumor and the local cells are capable of producing high levels of IL-2 and IFN-gamma.

We studied the microvasculature of the pancreatic islet with an electron microscope using 15 alloxan-diabetic and 16 control rats. These animals were morphologically examined at 2 weeks, 1, 2, and 4 months after alloxan injection (diabetic rats) or at comparable times (control rats). Control rats were non-diabetic, either sequentially treated with glucose and alloxan (injected controls) or not subjected to any medication (non-treated controls). Body weights, plasma glucose levels, and erythrocyte glycosylated hemoglobin (HbA1c) concentrations were also measured at intervals. All the diabetic rats exhibited retarded growth, hyperglycemia, and increases in HbA1c concentrations. The primary changes in diabetic islet capillaries can be grouped into four categories: (1) narrowing and closing of capillaries and occurrence of possible acellular capillaries, (2) formation of perivascular edema, (3) bulging and projection of endothelial cytoplasm, and (4) perivascular proliferation of collagen-like fibrils together with thickening of the basement membrane. These changes tended to worsen with diabetic duration. They are regarded as pathognomonic for diabetic microangiopathy. It is presumed that the present microvascular lesions of the islet play an important role in the pathogenesis and advancement of the diabetic state.

Methanol extracts of the green algaUlva pertusa contain four kinds of glycerolipids that are active as feeding-stimulants for marine herbivorous gastropods. These compounds are digalactosyldiacylglycerol (DGDG), 1,2-diacylglycerly-4'-O-(N,N,N-trimethyl)-homoserine (DGTH), 1-monoacylglyceryl-4'-O-(N,N,N-trimethyl)-homoserine (MGTH), and 6-sulfoquinovosyldiacylglycerol (SQDG). The various gastropods exhibit marked specificity, however, as young abaloneHaliotis discus respond to DGDG and DGTH at minute dosages of 20-30 μg/sample zone, but do not respond to 300 μg of SQDG, which is a phagostimulant for two other kinds of gastropods,Turbo comutus andOmphalius pfeifferi.

The effects of various methods for removal of monocytes and residual erythrocytes from human peripheral blood mononuclear cells (PBMC) on T cell subsets were examined. T cell subsets were determined in a fluorescence activated cell sorter (FACS) using fluorescein-conjugated monoclonal antibodies (anti-Leu-1, anti-Leu-2a and anti-Leu-3a antibodies). Removal of monocytes by methods based on adherence or phagocytosis decreased yields of lymphocytes and caused changes in the percentage and/or the fluorescence intensity of T cell subsets. Exclusion of monocytes from the FACS analysis by setting of the scatter gates was incomplete (about 80%). Removal of residual erythrocytes after Ficoll separation by ammonium chloride treatment also changed the percentage of T cell subsets. A method using PBMC without removal of monocytes and erythrocytes was chosen as best and simplest for routine use in FACS analysis of lymphocytes. Erythrocytes could be excluded from the FACS analysis by setting the scatter gates and the percentage of T cell subsets was corrected after measurement of monocytes, identified by peroxidase staining. The reproducibility of measurements of T cell subsets made at different times was examined using PBMC obtained from the same healthy man during 12 weeks. For standardization of the assay, the peak positions of scatter and fluorescence intensity of each PBMC labeled with anti-Leu-3a were adjusted to standard values in each FACS analysis. Under these conditions, variations of other parameters of this standard PBMC were very small in 12 different assays. Using this standard PBMC, satisfactory, reproducible results were also observed on PBMC obtained from another normal subject. Therefore, this standard PBMC labeled with anti-Leu-3a was used as a standard in FACS analysis. Under these accurately standardized conditions, it was demonstrated that the peak position of fluorescence intensity of Leu-2a (suppressor/cytotoxic T) cells was significantly lower (P less than 0.01) in women than in men.

A 69-year-old male visited our hospital in December, 1983, complaining of dysphasia. X-ray and endoscopic examination of the stomach revealed Borrmann's type 3 of the cardia. A biopsy specimen revealed typical features of adenocarcinoma. Because the tumor could not be resected, chemotherapy was started in February, 1984 with a combination of UFT (600 mg/day) and mitomycin (6 mg/week). After five courses of the combination chemotherapy, UFT therapy was continued. Follow-up endoscopy in August, 1985, showed a discolored area of the cardia. A biopsy specimen revealed no evidence of carcinoma. UFT-M therapy is a useful chemotherapy for gastric cancer.

It became increasingly evident that UFTM chemotherapy, a combination of UFT and mitomycin C is one of the most recommendable therapy for advanced gastric cancer, especially for the treatment of Borrmann type 4 gastric cancer. Objective responses were achieved in 18 out of 26 (69.2%) patients with Borrmann type 4 gastric cancer. Moreover, UFTM-Dpc treatment, a combination therapy of UFTM plus D-penicillamine (suppressor of collagen synthesis), is proposed for the treatment of Borrmann type 4 gastric cancer from the point of view of collagen synthesis inhibition. It is likely that D-penicillamine might inhibit the peritoneal metastatic involvements, which is lethal to the patients finally.

Investigations were made in human on the metabolism of Futraful (FT) and UFT (combination of uracil and FT), both of which are anticancer agents of pyrimidine metabolism antagonist. As a result, it was demonstrated that from the metabolic view point, those drugs essentially have a tumor-selective toxicity. Next, the mechanism of the superior clinical anticancer effect of UFT, compared to FT, was investigated enzymatically, showing the inhibitory effect of uracil on the degradation of 5-fluorouracil generated from FT in the organs and tumor tissues. Discussions were also made for the further development of fluoropyrimidines.

In brief, the clinical course of patients with Borrmann type 4 gastric cancer is characterized by its marked tendency for peritoneal metastasis (P-factor). This paper dealt with the treatment of Borrmann type 4 gastric cancer from the viewpoint of the disease P-factor, and included the following topics; UFTM chemotherapy, pharmacokinetics of mitomycin C (i.v. vs i.p. use), case presentation, Port-A-catheter (implantable drug delivery system), suppression of collagen biosynthesis, prevention of the adverse effects of mitomycin C, neochemotherapy, and autopsy findings.