Keishi Kojima

Fecal eosinophil granule-derived proteins reflect disease activity in inflammatory bowel disease

OBJECTIVES:The aims of this study were: 1) to examine whether the fecal levels of eosinophil granule-derived proteins reflect disease activity in inflammatory bowel disease (IBD); and 2) to examine the extracellular release of these proteins from eosinophils and their stability in feces by an in vitro study.METHODS:We investigated 42 patients with ulcerative colitis (UC), 37 patients with Crohns disease (CD), and 29 control subjects. The stool samples were collected at 4°C over 48 h and were homogenized. The fecal levels of eosinophil cationic protein (ECP) and eosinophil protein X (EPX) were measured by radioimmunoassay. Fecal Hb (Hb), α1-antitrypsin (AT), and lactoferrin (Lf) were also measured by ELISA.RESULTS:Fecal ECP and EPX concentrations were significantly increased in both active UC and active CD compared to inactive UC and inactive CD, respectively. Fecal EPX concentration correlated with the fecal Hb, AT, and Lf concentrations more closely than fecal ECP concentration. Even in the inactive stage, CD patients who relapsed within the following 3 months showed higher fecal ECP and EPX concentrations compared to the patients who did not. EPX was released extracellularly more efficiently than ECP (18.6%vs 6.3%, after incubation for 15 min at 25°C). EPX was more stable in the feces than ECP.CONCLUSIONS:The measurement of eosinophil granule-derived proteins in feces is useful for evaluating disease activity and predicting relapse in patients with IBD. EPX may be more suitable than ECP as a fecal eosinophil marker.

Lactoferrin in whole gut lavage fluid as a marker for disease activity in inflammatory bowel disease: comparison with other neutrophil-derived proteins

OBJECTIVES:We investigated which neutrophil-derived proteins in whole gut lavage fluid (WGLF) most accurately reflect disease activity in inflammatory bowel disease.METHODS:WGLF was obtained from patients undergoing whole gut lavage as a bowel preparation for colonoscopy. Twenty-seven patients with ulcerative colitis (UC), 23 patients with Crohns disease (CD), and 35 control subjects were examined. The concentrations of lactoferrin, polymorphonuclear neutrophil elastase (PMN-E), myeloperoxidase, and lysozyme in WGLF were measured by ELISA. For the assessment of stability, WGLF samples were stored at 37°C for various periods.RESULTS:In UC, the concentrations of lactoferrin, myeloperoxidase, and lysozyme in WGLF had good correlations with colonoscopic grading. Zero, 12, five, and 10 of 28 samples from active UC patients showed normal concentrations of lactoferrin, PMN-E, myeloperoxidase, and lysozyme, respectively. In CD, the concentrations of lactoferrin and myeloperoxidase had good correlations with the Crohns disease activity index. Thirteen and seven of 36 samples from inactive CD patients (Crohns disease activity index ≤ 150) showed high concentrations of lactoferrin and myeloperoxidase, respectively. Most of them (11/13, 6/7) were found to have ulceration by colonoscopy or small bowel x-ray. The ratio of the lactoferrin concentration in the WGLF supernatant to that in total WGLF was highest among these proteins in all disease groups and control subjects. Lactoferrin and myeloperoxidase showed good stability in WGLF, whereas PMN-E and lysozyme did not.CONCLUSION:Lactoferrin is the most suitable of these proteins for use as a neutrophil-derived WGLF marker of intestinal inflammation.

Antineutrophil cytoplasmic antibodies in Japanese patients with inflammatory bowel disease: prevalence and recognition of putative antigens

Objective:Our aim was to investigate the prevalence of antineutrophil cytoplasmic antibodies (ANCA) in Japanese patients with ulcerative colitis (UC) and Crohns disease (CD), and the putative antigens recognized by perinuclear staining pattern ANCA (p-ANCA)–positive sera.Methods:Sera from UC n = 52 and CD n = 43 patients, and from healthy controls n = 74 were studied. The indirect immunofluorescence (IIF) method was used for the detection of ANCA and its binding pattern. p-ANCA–positive sera were studied further for putative antigens. ELISAs using lactoferrin (Lf), myeloperoxidase (MPO), and cathepsin G (Cat G) as antigens were performed.Results:ANCA was positive in 40 of the 52 (76.9%) UC (p-ANCA in 33) and in 32 of the 43 (74.4%) CD (p-ANCA in 31) patients. UC and CD patients showed significantly higher titers of p-ANCA than controls; however, no significant difference was observed between UC and CD. In UC, 23, 17, and nine of the 33 patients with p-ANCA–positive sera showed reactivity with Lf, MPO, and Cat-G, respectively. In CD, 21, 20, and 11 of the 31 patients with p-ANCA–positive sera showed reactivity with Lf, MPO, and Cat-G, respectively. Fourteen of the UC and six of the CD patients showed reactivity with two different antigens, and seven of the UC and 11 of the CD patients showed reactivity with all three antigens. The presence of anti-Lf and anti-MPO antibodies was further confirmed by Western blotting.Conclusions:ANCA is useful in distinguishing patients with IBD from normal subjects but is not sufficient for the differential diagnosis of CD and UC. p-ANCA reactivity might be derived from the recognition of heterogeneous neutrophil-associated antigens.

Medium-chain fatty acids stimulate interleukin-8 production in Caco-2 cells with different mechanisms from long-chain fatty acids

Background and Aim: It has been suggested that dietary fat exacerbates intestinal inflammation. We investigated the effect of fatty acids on interleukin (IL)‐8 production in a human intestinal epithelial cell line (Caco‐2).

Cyclosporine A inhibits interleukin-8 production in a human colon epithelial cell line (HT-29)

Intestinal epithelial cells produce various inflammatory mediators. However, the way in which immunosuppressive agents influence the production of these mediators by intestinal epithelial cells is not understood. The effects of cyclosporine A (CsA), tacrolimus (FK506), and dexamethasone (DEX) on cytokin-induced production of interleukin (IL)-8 in a human colonic cancer cell line (HT-29) were examined. HT-29 cells were stimulated with either IL-1β or tumor necrosis factor α (TNFα) together with CsA, FK506, or DEX. The presence of IL-8 protein was detected by enzyme-linked immunosorbent assay, and the expression of IL-8 messenger RNA (mRNA) by reversetranscription polymerase chain reaction. CsA (1, 5, and 10ng/ml) significantly reduced IL-1β-induced IL-8 production (by 32%, 41%, and 48%, respectively), and reduced TNFα-induced IL-8 production (by 21%, 42%, and 50%, respectively). FK506 or DEX had no effect on IL-1β- or TNFα-induced IL-8 production. The expression of IL-8 mRNA was also inhibited by CsA. These findings suggest that CsA may influence the production of inflammatory mediators in colonic cells in a different manner from FK506 and DEX.

Bile acids inhibit tumour necrosis factor α-induced interleukin-8 production in human colon epithelial cells

To clarify the regulatory mechanism of the production of various inflammatory mediators by intestinal epithelial cells, the effect of bile acids (tauroursodeoxycholate, TUDC; taurochenodeoxycholate, TCDC; and taurocholate, TC) on the cytokine‐induced production of interleukin (IL)‐8 in a human colon epithelial cell line (HT‐29) was examined. HT‐29 cells were incubated for 24 h in a culture medium containing tumour necrosis factor α (TNFα; 1 ng/mL) and/or interleukin (IL)‐1 β (1 ng/mL) in the presence or absence of bile acids. The IL‐8 concentration in the medium was measured by an enzyme‐linked immunosorbent assay. The binding assay of TNFα was performed using [125I]‐TNFα (100 pmol/L). Interleukin‐8 production during incubation with TNFα was markedly reduced in the presence of 0.5 and 1 mmol/L TUDC, 0.5 and 1 mmol/L TCDC and 0.5 and 1 mmol/L TC, by 56, 85, 86, 91, 37 and 70%, respectively. The IL‐8 production during incubation with IL‐1ß was not significantly reduced in the presence of these bile acids. The specific binding of TNFα to cells was inhibited 33, 47, and 14% by 1 mmol/L TUDC, TCDC and TC, respectively. These findings suggest that bile acids inhibit TNFα‐induced IL‐8 production by the colonic cells. The suppression may be partly due to inhibition of TNFα binding to the cells by bile acids.

Original ContributionsAntineutrophil cytoplasmic antibodies in Japanese patients with inflammatory bowel disease: prevalence and recognition of putative antigens

OBJECTIVE: Our aim was to investigate the prevalence of antineutrophil cytoplasmic antibodies (ANCA) in Japanese patients with ulcerative colitis (UC) and Crohn’s disease (CD), and the putative antigens recognized by perinuclear staining pattern ANCA (p-ANCA)–positive sera. METHODS: Sera from UC (n = 52) and CD (n = 43) patients, and from healthy controls (n = 74) were studied. The indirect immunofluorescence (IIF) method was used for the detection of ANCA and its binding pattern. p-ANCA–positive sera were studied further for putative antigens. ELISAs using lactoferrin (Lf), myeloperoxidase (MPO), and cathepsin G (Cat G) as antigens were performed. RESULTS: ANCA was positive in 40 of the 52 (76.9%) UC (p-ANCA in 33) and in 32 of the 43 (74.4%) CD (p-ANCA in 31) patients. UC and CD patients showed significantly higher titers of p-ANCA than controls; however, no significant difference was observed between UC and CD. In UC, 23, 17, and nine of the 33 patients with p-ANCA–positive sera showed reactivity with Lf, MPO, and Cat-G, respectively. In CD, 21, 20, and 11 of the 31 patients with p-ANCA–positive sera showed reactivity with Lf, MPO, and Cat-G, respectively. Fourteen of the UC and six of the CD patients showed reactivity with two different antigens, and seven of the UC and 11 of the CD patients showed reactivity with all three antigens. The presence of anti-Lf and anti-MPO antibodies was further confirmed by Western blotting. CONCLUSIONS: ANCA is useful in distinguishing patients with IBD from normal subjects but is not sufficient for the differential diagnosis of CD and UC. p-ANCA reactivity might be derived from the recognition of heterogeneous neutrophil-associated antigens.

Therapy of GERD and FD overlap with symptoms after usual-dose PPI: acotiamide plus usual-dose PPI vs. double-dose PPI

Gastroesophageal reflux disease (GERD) and functional dyspepsia (FD) often coexist or overlap. In this study, the efficacy of acotiamide in combination with a standard dose of rabeprazole for GERD and FD was compared with that of a double dose of rabeprazole.

Does the Novel Potassium-Competitive Acid Blocker Vonoprazan Cause More Hypergastrinemia than Conventional Proton Pump Inhibitors? A Multicenter Prospective Cross-Sectional Study

Background/Aim: The long-term administration of proton pump inhibitors (PPIs) is useful for preventing recurrent reflux esophagitis. On the other hand, several adverse reactions, such as an increase in the blood gastrin level, have been reported. The aim of the present study was to examine the increase in the blood gastrin level due to the long-term administration of conventional PPIs compared with vonoprazan. Methods: A prospective cross-sectional study was conducted. We examined the blood gastrin levels of patients taking vonoprazan or conventional PPIs in whom the grade of atrophic gastritis had been endoscopically evaluated in the last year. Results: The blood gastrin level was significantly higher in the vonoprazan group than that in the PPI group in patients with milder or no atrophic gastritis, irrespective of the administration periods. However, no significant difference was observed between the groups in patients with severe atrophic gastritis. Conclusion: Vonoprazan more markedly increased the blood gastrin level compared with conventional PPIs in patients with milder or no atrophic gastritis. This indicates that vonoprazan may have stronger acid-suppressing effects in such patients than conventional PPIs. Key Message: We should be aware of the potential development of hypergastrinemia during the long-term administration of vonoprazan, especially in patients with mild or no atrophic gastritis.

Endoscopic Submucosal Dissection Techniques for Early Gastric Cancer

Endoscopic Submucosal Dissection Techniques for Early Gastric Cancer Eiji Umegaki, Satoshi Tokioka, Masaya Tanaka, Nozomi Takeuchi, Chikako Eiraku, Takao Noguchi, Minekazu Ozawa, Toshihisa Takeuchi, Nanako Shiraishi, Keishi Kojima, Ichiro Hirata, Ken-ichi Katsu Although the strip biopsy method is popular endoscopic mucosal resection technique (EMR) for its convenience and reliability, it has limitations in resectable tumor size. Endoscopic submucosal dissection techniques (ESD) using the diathermic needle knife or the insurated-tip diathermic knife have been introduced to overcome this disadvantage, but they have high risks for bleeding and perforation. Our intent in this study was to investigate the usefulness of ESD for early gastric cancer comparison with the strip biopsy method and the educational system in our hospital. Materials and Methods: Studies were carried out on 505 lesions in whom EMR was performed, 385 lesions were treated with the strip biopsy and 120 with ESD. We investigated the en-bloc resection rate, the complication rate of bleeding and perforation, and the learning curve of ESD in our hospital. Results: (1) The en-bloc resection rate of the strip biopsy method was 54.8% and that of ESD was 87.4%. (2) Irrespective of tumor size and location, we could resect the tumor with a much higher en-bock resection rate. (3) The complication rate of bleeding and perforation was 0.7% in the strip biopsy method and 8.7% in ESD. (4) Training for ESD was acquired in an informal setting (observation of actual procedures by videotapes, animal models) and a formal setting. Nothing can replace live demonstrations of ESD. (5) The learning curve of ESD in our hospital. Conclusion: Endoscopists who seek to perform ESD should avail themselves of training and education in a formal or informal setting. ESD is promising as a safe and reliable technique for the treatment of early gastric cancer. T1402 Endoscopic Gastroenteric Anastomoses with Magnets (EGAM): Three Years After Augusto Villaverde, Constantine Cope, Nestor Chopita, Nestor Landoni, Alberto Bernedo, Horacio Martinez, Alejandro Jmelnitzky Introduction: Three years ago we presented a novel technique of gastroenteric anastomoses, formed by endoscopic magnet assisted nonsurgical technique as an alternative to enteral stents(SEMS) and surgery for malignancies involving the duodenum or gastric antrum. Aims & Methods: The purpose of this study is to evaluate three years of work with this technique and evaluate early and late complication rates. Patients: From December 2001 until November 2004, 21 patients with malignant biliary, gastric and duodenal obstruction were included in the study. All patients were deemed non-surgical candidates because of their advanced disease and poor performance status, according to AJCC classification. Methods: All patients had biliary drainage performed prior to EGAM either by ERC or PTC. Once the EGAM was completed, the patients were included in a monthly follow-up protocol with endoscopy, radiology, evaluation of symptoms, nutritional status, and the Karnoftsky rating scale. Results: The procedure was successful in 19 of 21 patients. All patients started with oral intake after one day of the procedure and were hospitalized for two days. Early complications was perforation of immature fistula in two patients. During the follow-up, 3 stent migrations and 1 obstruction of the stent by solid food, were presented. Ten patients maintained their baseline Karnofsky score, two patients increased this by one point and 1 patient increased this score by 2 points and is still alive. All patients had good patency of the anastomoses until their time of death. The mean survival was 4.3 months. Conclusion: Our results demonstrate the safety and efficacy of EGAM as a novel, non-surgical option for creating a gastroenteric anastomosis in human beings. We believe this minimally invasive technique is a valid alternative to SEMS with good results in long term follow up. More trials are necessary to confirm our findings.