Mucosal proinflammatory cytokine and chemokine expression of gastroduodenal lesions in Crohn's disease.

BACKGROUND: The stomach and the duodenum are frequent sites of involvement by diminutive lesions in Crohn's disease (CD). AIM: To assess mucosal proinflammatory cytokines and chemokines in gastroduodenal lesions of CD. METHODS: 13C-Urea breath test and upper endoscopy were performed in 29 CD patients and seven control subjects, and biopsy specimens were obtained from the gastric cardia and the duodenum. Histology and mucosal levels of IL-1beta, IL-8/CXCL8 and RANTES/CCL5 were assessed and compared according to the presence of gastric cardial lesion [bamboo joint-like appearance (BJA)] and duodenal lesion (notched appearance, aphthous erosion and polypoid lesion). In 11 CD patients, these procedures were repeatedly performed after administration of famotidine. RESULTS: H. pylori was less frequently positive in CD patients than in controls (10% vs. 71%, P = 0.003). Prevalence of cardial and duodenal lesion was significantly higher in CD than in controls (59% vs. 0%, P = 0.008 for gastric lesion; 45% vs. 0%, P = 0.034 for duodenal lesion). There were no differences in IL-1beta, IL-8 and RANTES between CD and controls. Duodenal mucosal IL-1beta and IL-8 were significantly higher in positive duodenal lesion than in negative duodenal lesion. However, there were no such differences with respect to cardial lesions. Endoscopic findings remained unchanged after administration of famotidine, while there was a trend towards decreases in IL-1beta and IL-8 in the gastric cardia. CONCLUSIONS: The pathogenesis of diminutive lesions of CD may be different between the stomach and the duodenum. Famotidine may not have a therapeutic effect on duodenal lesion in CD.

Small invasive colonic cancer occurring in a hyperplastic polyp.

We report a case of advanced colon cancer which was supposed to have arisen from a hyperplastic polyp in a 68-year-old man. Colonoscopy revealed a depressed reddish area with a surrounding elevated lesion that was of a faded color compared with the normal mucosa. After the mucosal surface had been sprayed with crystal violet dye, magnifying colonoscopy showed an amorphous area in the central depression and the surrounding, slightly elevated lesion had an asteroid pattern. The depressed area was therefore considered to be a colonic cancer surrounded by a hyperplastic polyp. Endoscopic ultrasonography showed that the lesion was infiltrating further than the deep submucosal layer and it was therefore decided to treat the patient by laparoscopically assisted right hemicolectomy. The depressed lesion was found to be a well-differentiated adenocarcinoma invading the muscularis propria (diagnosed as IIc + IIa-like advanced adenocarcinoma). The surrounding flat elevated lesion was found to be hyperplastic mucosa. No adenomatous lesions were found. There have been few reported cases in which a preoperative diagnosis of carcinoma in a hyperplastic polyp has been made, but the possibility of carcinogenesis from hyperplastic polyps has come under consideration recently. This case was considered to be important because it raises the possibility that nonpolypoid cancer can develop from a hyperplastic polyp.

CCR4 is an up-regulated chemokine receptor of peripheral blood memory CD4+ T cells in Crohn's disease.

Several chemokine receptors are expressed selectively on the surface of T cells depending on their polarization. The aim of this study was to characterize chemokine receptor expression in peripheral blood memory T cells in Crohn's disease (CD) and ulcerative colitis (UC), and to correlate the expression with disease activity. Peripheral blood mononuclear cells (PBMCs) were obtained from 24 patients with CD, 30 patients with UC, 24 normal controls and 10 disease controls. PBMCs were stained by anti-CCR3, CCR4, CCR5, CXCR3, CD4, CD8, CD45RO and beta 7 integrin, and the expression of the chemokine receptors were determined by flow cytometry. CCR4 expression on memory T cells was significantly lower in UC than in CD or normal controls, and that of memory CD4+ T and beta 7(high) memory CD4+ T cells was significantly higher in CD than in UC or normal controls. CCR4 expression on memory CD4+ T cells exhibited significant positive correlation with disease activity in CD, and this decreased significantly after treatment. Such a decrease was not found in the disease controls. CCR5 and CXCR3 expression on memory CD8+ T cells was significantly lower in CD than in normal controls. CXCR3 expression on beta 7(high) memory CD4+ T and CXCR3 expression on memory CD8+ T cells were lower in UC than in normal controls. These findings suggest that in peripheral blood memory T cells, chemokine receptor expression is different between CD and UC. Enhancement of CCR4 and suppression of CCR5 and CXCR3 seem to be the characteristic chemokine receptor profile in peripheral blood memory T cells of CD.

Permeation of oxygen, water vapor, and limonene through printed and unprinted biaxially oriented polypropylene films.

Oriented polypropylene (OPP) and coated OPP (acrylic/OPP/PVDC) films were printed with two commercially available inks to investigate the influence of inks on water vapor and oxygen transmission rates. The permeation of an aroma compound (d-limonene) through coated OPP film printed with these inks was also evaluated at 35 degrees C and 100% relative humidity. The water vapor transmission rate increased significantly through OPP film printed with nitrocellulose-based ink. The oxygen transmission rate was significantly lower through both OPP and coated OPP films printed with the nitrocellulose ink. The effect of inks on limonene permeation was minor compared to the marked increase in permeation measured when the PVDC side of the coated film was exposed to the aroma, compared to the acrylic side. Scanning electron micrographs of coated film cross sections revealed changes in film structure upon exposure to limonene vapors, which were most pronounced when the PVDC side was exposed to limonene.

IL-7 prevents both caspase-dependent and -independent pathways that lead to the spontaneous apoptosis of i-IEL.

Intestinal intraepithelial lymphocytes (i-IEL) readily undergo spontaneous apoptosis in vitro through an unclear mechanism. Here we examined the relationship between caspases, which plays a major role in apoptosis, and IL-7 in the spontaneous apoptosis of i-IEL in vitro. We demonstrated that IL-7 and zVAD prevented the spontaneous apoptosis of i-IEL by approximately 50% and 25% respectively with no additive protection seen when both are used. IL-7 preferentially prevented the apoptosis of gammadelta i-IEL, while zVAD equally prevented the apoptosis of gammadelta and alphabeta i-IEL. Lastly, we demonstrated that the spontaneous apoptosis of i-IEL is associated with a marked increase in caspase activity. Caspase activity was completely inhibited by zVAD, but only slightly by IL-7. Overall these results suggest that two pathways lead to the spontaneous apoptosis of i-IEL, one which is caspase dependent and the other which is caspase independent. IL-7 appears to exert its effect on i-IEL undergoing spontaneous by partially inhibiting both apoptotic pathways.

Clinical study of strangulation obstruction of the small bowel.

Early diagnosis of strangulation obstruction is very important for surgeons because delayed diagnosis often leads to severe complications. Thirty patients underwent an operation because of small bowel obstruction between April, 1993 and December, 1999. In the present study, we examined the differences in clinical findings between simple obstruction and strangulation obstruction. In addition, we examined the manifestation of systemic inflammatory response syndrome (SIRS) and whether it is useful for early diagnosis of strangulation obstruction, and whether it is correlated with the severity of ischemia due to strangulation. Tenderness was examined in all patients and signs such as abdominal irritation were detected more often in patients with strangulation obstruction than in the patients with simple obstruction. According to SIRS, the large number of the patients with strangulation obstruction showed SIRS before operation and the manifestation of SIRS correlated well with the length of the necrosis in the strangulated small bowel. We recognized the importance of anamnesis and clinical findings in examinations of small bowel obstruction, furthermore, it was suggested that SIRS should be the warning sign for strangulation obstruction.

p53-dependent radiation-induced crypt intestinal epithelial cells apoptosis is mediated in part through TNF-TNFR1 system.

Radiation induces apoptosis of crypt intestinal epithelial cells (IEC) through a pathway that is largely dependent on p53. However, exactly how p53 mediates IEC apoptosis is unclear. Studies in vitro suggest that one mechanism by which p53 mediates apoptosis is through its ability to transactivate members of the TNF receptor family of 'Death Receptors'. Here, we examined the role of one of its member, TNF receptor type 1 (TNFR1), in an in vivo model of p53-dependent radiation-induced IEC apoptosis. We demonstrate that mice genetically engineered to be deficient in TNF receptor type 1 (TNFR1(-/-)) and mice injected with TNFR1-fusion chimeric protein (TNFR1-Fc; a competitive inhibitor of TNFR1) were partially protected (30-40%) from p53-dependent radiation-induced IEC apoptosis. However, we found no evidence to support the possibility p53 transcriptionally regulates the expression of TNFR1 nor increases the susceptibility of IEC to TNF-mediated apoptosis. Interestingly, we found that injection of TNF readily induced IEC apoptosis and that radiation induced a p53-dependent increase in the intestinal level of TNF. Furthermore, injection of a neutralizing anti-TNF mAb reduced p53-dependent radiation-induced IEC apoptosis by approximately 60%. Overall, these results suggest that p53-dependent radiation-induced IEC apoptosis is mediated in part through ability of p53 to regulate TNF, which subsequently induces IEC apoptosis through TNFR1.

The effect of interleukin-4 on the induction of intestinal mast cells and chronological cytokine profiles during intestinal nematode Strongyloides ratti infection.

The effect of interleukin-4 (IL-4) on the induction of intestinal mast cells and cytokine profiles during Strongyloides ratti infection was studied using IL-4 knockout (IL-4 KO) mice. The antigen-specific proliferative response of mesenteric lymph node cells was not impaired in IL-4 KO mice. The number of intestinal mast cells induced in IL-4 KO mice during S. ratti infection was 2- to 3-fold lower than that observed in WT mice. Intestinal mastocytosis had disappeared in IL-4 KO mice by day 21 postinfection, when significant mastocytosis continued to be observed in WT mice. In mesenteric lymphnode of IL-4 KO, IL-3 production decreased and mice IFN-gamma production significantly increased as compared with those of WT mice. The numbers of eggs excreted per gram of feces (EPG) by IL-4 KO mice were greater than those excreted by WT mice on day 6 postinfection, but no difference was observed in the subsequent period. In conclusion, intestinal mast cells are induced during S. ratti infection in the absence of IL-4, and IL-4 is not essential for protection against intestinal adult worms of S. ratti.

Well-differentiated adenocarcinoma of the gallbladder mimicking minimal deviation adenocarcinoma of the cervix.

We report a case of well-differentiated adenocarcinoma of the gallbladder, histologically mimicking minimal deviation adenocarcinoma (MDA) of the cervix. A 71-year-old Japanese male underwent cholecystectomy because of the suggestion of gallbladder carcinoma. The resected gallbladder showed a localized thickening of the gallbladder wall with a polypoid lesion measuring 12x7 mm in diameter. Microscopically, the polypoid lesion proved to be a well-differentiated adenocarcinoma composed of columnar cells with a clear cytoplasm. In the thickened gallbladder wall, well-formed glands were extensively distributed; they were surrounded by a slightly desmoplastic reaction instead of lamina propria, or were directly in contact with smooth muscle cells. The diagnostic criteria for cervical MDA may be useful in distinguishing well-differentiated adenocarcinoma of the gallbladder from benign conditions, such as Rokitansky-Aschoff sinus and adenomyomatosis. It is remarkable that the tumor cells of the present case expressed gastric type mucin which is characteristic of mucinous type cervical MDA.

Differential requirements of CD45 protein tyrosine phosphatase for cytolytic activities and intrathymic and extrathymic development of intestinal intraepithelial lymphocytes.

CD45 is a transmembrane protein tyrosine phosphatase essential for Ag receptor-mediated signaling in both T and B cells. In this study we investigated roles of CD45 in development and cytolytic activities of murine intestinal intraepithelial lymphocytes (i-IEL) using CD45 exon 6 knockout (CD45-/-) mice. Interestingly, the total cell number of i-IEL was significantly reduced in CD45-/-mice during aging (10-20 wk of age), whereas the i-IEL number was normally increased in the wild-type littermates. Especially, the number of gamma(delta)TCR+ i-IEL decreased markedly in CD45-/- mice during aging. The i-IEL in CD45-/- mice were more susceptible to in vitro spontaneous apoptosis than the normal i-IEL, implying that CD45 is required for maintenance of the cellularity of i-IEL. Results from in vivo analyses of the extrathymic and intrathymic development of i-IEL suggested that CD45-mediated signaling is required for the intrathymic, but not the extrathymic, development of i-IEL. Moreover, the whole i-IEL from CD45-/- mice showed a significantly reduced cytolytic activity, and the residual cytolytic activity was completely diminished by depleting CD45+ i-IEL, suggesting that CD45 is indispensable for the TCR-mediated cytolytic activity of i-IEL. Furthermore, we found differential contributions of CD45 and p56lck to development and induction of cytolytic activities of i-IEL.

Case report of an anomalous arrangement of the pancreaticobiliary ducts and nuclear DNA ploidy analysis.

Two cases with an anomalous arrangement of the pancreaticobiliary ducts (APBD) are reported in order to investigate the mechanism of carcinogenesis of the extrahepatic biliary tract from the aspect of DNA ploidy analysis. Highly elevated biliary enzymes were found in both cases. Although inflammatory changes without any metaplasia or neoplasia were observed histologically, a cytofluorometric nuclear DNA ploidy analysis showed an aneuploid low ploidy pattern in a histogram of the gall bladder, an aneuploid high ploidy pattern in a histogram of the common bile duct in one patient, and a diploidy pattern in the gall bladder, an aneuploid low pattern in the common bile duct in another case. These results may show that APBD may play a role in the development in DNA ploidy abnormality with refluxed pancreatic juice which may induce repeated irritation and inflammation. Radical surgery including a complete excision of the extrahepatic biliary tract followed by reconstruction may achieve better results with regard to undesirable consequences with an abnormal nuclear DNA ploidy pattern and decrease the likelihood of developing carcinoma.

[Effect of synthetic protease inhibitor on the sphincter of Oddi function in dogs].

Fundamental study in dogs have shown that the synthetic protease inhibitor has pharmaceutical properties characterized by effect on motility of the sphincter of Oddi. Synthetic protease inhibitors, gabexate mesilate and nafamostat mesilate have effects on motility of the sphincter of Oddi in dogs. The motor effect of synthetic protease inhibitor on the sphincter of Oddi has been investigated by manometric evaluation. Immediately after intravenous administration of gabexate mesilate (5, 10, 50, and 100 mg/kg/h) the motility of the sphincter of Oddi was inhibited dose-dependently, on the other hand after intravenous administration of nafamostat mesilate (0.5, 1, and 5 mg/kg/h) motility of the sphincter of Oddi was accelerated. Therefore it was suggested that these results shown some considerable problems in clinical use.

Biliary stones and bacteriae in bile study in 211 consecutive cases.

A retrospective study have been performed to investigate the correlation between stone formation and bacteriae in the bile of 211 consecutive biliary stone patients (133 females and 78 males, age ranged 1.3 to 87.0 years old, 196 cholecystolithiasis, 54 choledocholithiasis and 9 hepatolithiasis). They had undergone surgery since January 1980 to December 1987. Bile was collected to perform bacterial culture from the gallbladder and common bile duct during operation and all stones had been classified according to morphology after surgery and confirmed with component analysis if necessary. Totally 5115 stones had been collected and 20.4% had been classified as pure cholesterol stone, 16.8% as combination stone, 17.2% as mixed stone, 23.6% as calcium bilirubinate stone, 17.6% as black stone, 0.8% as calcium fatty acid stone and 3.6% as unknown. In bile culture, 84 from the gallbladder and 31 from the common bile duct were positive. Streptococci were the most frequent and followed by Klebsiella, Escherichia coli and so on. A higher incidence of positive culture was shown in calcium bilirubinate and lower incidence was shown in cholesterol stones. These results suggest that bacteriae in the bile may play a role in the formation of pigment biliary stones.

[Effects of cholecystokinin-octapeptide and tetragastrin on the contractile activities of the stomach, duodenum and papilla of Vater].

Effects of cholecystokinin-octapeptide (CCK-OP) and tetragastrin (TG) on the stomach, duodenum and papilla of Vater were examined using 13 adult dogs. The contractile activities of the stomach and duodenum were examined using the strain gage, and the manometric study of the papilla of Vater was also performed. CCK-OP increased the contractile activities of the stomach and duodenum, but at the same time the pressure of the papilla of Vater was decreased. TG increased the contractile activities of the stomach, but decreased that of the duodenum and the pressure of papilla of Vater. CCK-OP lowered the pressure of the common bile duct, and that fall corresponded with the disappearance of the phasic contraction of the papilla of Vater. We concluded that the contractile activity of the papilla of Vater was independent of the contractile activity of the duodenum, and the most effective drainage of the bile was achieved at the time of the disappearance of the phasic contraction of the papilla of Vater.

Long-term storage of blood samples at freezing temperatures in the presence of a cryoprotectant for hemiglobin assay.

Changes in Hi levels in experimentally prepared blood samples during storage at various temperatures were studied. When whole blood in which Hi levels were elevated by sodium nitrite was stored unfrozen, rapid reduction of Hi was observed within 24 hr even at 0 degrees C. When whole blood or a diluted hemolysate was stored frozen for a week or longer, considerable formation of Hi by autoxidation was observed, the formation at -20 degrees C being much more significant than that at -30 degrees C. On the other hand, addition of an equal volume of the cryoprotectant solution of Rowe et al. to blood almost completely inhibited this Hi formation during freezing storage until at least 30 days. Thus, a new method for long-term storage of blood samples for Hi assay was devised.

Identification of fetal bloodstains by enzyme-linked immunosorbent assay for human alpha-fetoprotein.

A rapid and highly sensitive enzyme-linked immunosorbent assay (ELISA) for determination of human alpha-fetoprotein (AFP) using commercially available reagents was devised and applied to identification of fetal bloodstains. When experimentally prepared bloodstains, 1 by 2 mm in area, were submitted to analysis, only fetal bloodstains showed positive reactions in the present ELISA. The reactions did not change significantly when these bloodstains were stored at room temperature for one week. The present ELISA seems to be suitable for forensic science practice.