Masamichi Satomi

Multicenter Randomized Controlled Trial for the Treatment of Ulcerative Colitis with a Leukocytapheresis Column

The administration of steroids is not always effective for the treatment of ulcerative colitis (UC). Their long-term use often causes adverse effects which sometimes result in their stoppage and acute exacerbation. Therefore, an alternative treatment is necessary in order to decrease steroid dosage and avoid the clinical problems associated with steroids. Methods The effectiveness and adverse effects of a leukocytapheresis (LCAP) were investigated in a controlled multicenter trial with randomized assignment of 76 active-stage UC patients in two groups. In the LCAP group (39 patients), LCAP weekly for 5 weeks as an intensive therapy was added to the on-going drug therapy, while steroids were maintained but not increased, and then LCAP was gradually reduced to once every 4 weeks as a maintenance therapy. In the high dose prednisolone (h-PSL) group (37 patients), PSL was added or increased 30 approximately 40 mg/day for moderately severe and 60 approximately 80 mg/day for severe patients and then gradually tapered. Findings The LCAP group showed a significantly higher effectiveness (74% vs. 38%; p=0.005) and lower incidence of adverse effects (24% vs. 68%; p<0.001). The patients were able to continue the trial for a longer period in the LCAP group than the h-PSL group (p=0.012). Clinical activity and endoscopic indexes showed the LCAP group had better improvements than the h-PSL group. Interpretation The results of the trial show that LCAP permits a reduction in total PSL dosage and is more effective and safer than high-dose PSL administration for intensive therapy, and LCAP may maintain remission longer than PSL.

Leukocytapheresis therapy, performed with leukocyte removal filter, for inflammatory bowel disease

Leukocytapheresis (LCAP), performed with a leukocyte removal filter, was administered five times, at 1-week intervals, for 5 weeks of intensive therapy and five times, at approximately 1-month intervals, for approximately 5 months of maintenance therapy, to 13 patients with inflammatory bowel disease (IBD) diagnosed as ulcerative colitis (UC) in 8 and Crohns disease (CD) in 5. Clinical and blood examinations showed no side effects in any of the patients. During the intensive therapy, excellent or moderate clinical response was recognized in 11 of the 13 patients (84.6%), of whom 6 had a dramatic response; the excellent or moderate clinical response continued throughout the maintenance therapy in 8 of the patients (61.5%). Flow cytometry showed that the patients who had improved generally had high values for percentages of HLADR+, HLADR+CD3+, and HLADR+CD8+ cells before the first LCAP, and that these values and the C-reactive protein levels and erythrocyte sedimentation rates had decreased to the normal range by the end of both intensive and maintenance therapy. In the patients who showed poor response, in contrast, all the above values had been at or near normal before the initial LCAP administration. The clinical improvement in the absence of any additional medical treatment suggests that LCAP has the capacity to influence the causal mechanism(s) of IBD and that IBD is strongly associated with the cell-mediated immune response.

IL18 polymorphism is associated with an increased risk of Crohn’s disease

Background. The etiology of inflammatory bowel disease, which includes ulcerative colitis and Crohn’s disease, has not yet been made clear. However, inflammatory bowel disease is recognized as a multifactorial disease, and innate genetic factors might contribute to the pathogenesis. Cytokine genes are thought to be important in inflammatory bowel disease. Recently, interleukin 18, cloned as a novel proinflammatory cytokine, has been implicated in inflammatory bowel disease, especially Crohn’s disease. Methods. To identify germline mutations in patients with inflammatory bowel disease, the entire coding region of IL18 was examined using a DNA sequencing procedure. Results. No functional mutations were found, but a novel single nucleotide polymorphism (SNP) was identified as TCA/ TCC at codon 35. In patients with Crohn’s disease, the frequency of TCC allele carriers was significantly higher than in healthy controls (χ2 = 9.35, P = 0.002229, OR = 2.58, 95% CI = 1.39–4.80). Also, the magnitude of the association was more remarkable in females (χ2 = 16.36, P = 0.000052, OR = 8.17, 95% CI = 2.73–24.41). The TCC allele at codon 35 of IL18 may increase the risk for Crohn’s disease, especially in females. Conclusions.IL18 is probably one of several genes that determine susceptibility to Crohn’s disease.

Helicobacter pylori Infection Increases Mucosal Permeability of the Stomach and Intestine

It is important to study the effect of Helicobacter pylori infection on the permeability of the intestine. Permeability was evaluated by oral sucrose tolerance test using sucrose 25 g in 200 ml of water. Existence of H. pylori itself was associated with increased permeability of sucrose. Also, the permeability of sucrose increased as polymorphonuclear and lymphocyte infiltration increased. The increase of mucosal permeability suggests that antigens like protein penetrate into the body and result in systemic reactions. Thus, it is important to study the implication of increased permeability in relation not only to gastric diseases but also certain systemic diseases.

Fecal bile acid analysis in healthy Japanese subjects using a lipophilic anion exchanger, capillary column gas chromatography and mass spectrometry

SummaryDetailed fecal bile acid profiles of healthy Japanese subjects were studied using a lipophilic anion exchanger, capillary gas chromatography and mass spectrometry. Total daily excretion of bile acid into feces corrected for by fecal markers were between 127.99 to 366.33 μmole per day. Unconjugated bile acids constituted a major part, between 80 to 96%, of fecal bile acids. Glycine conjugated, taurine conjugated and sulfated bile acids were between 1 to 6, 0 to 3 and 1 to 10%, respectively. Esterified bile acids at C-24 position existed between 1 to 5%. Primary bile acids ranged from 0 to 55%. There were a number of epimers of hydroxy-and keto-bile acids, and lithocholic and deoxycholic acid were major secondary bile acids among them. A cholenoic acid was detected in the unconjugated fraction of one subject. It seems necessary to analyze the details not only on the type of bile acids but also on the mode of conjugation in biological samples. Thus, the methodology described in this study has made it easier to investigate on the role of bile acid in the physiology or pathophysiology of the gastrointestinal tract.

Infiltration of peroxidase-producing eosinophils into the lamina propria of patients with ulcerative colitis

Abstract: Little information is available to explain the pathogenesis of ulcerative colitis (UC). In this study, we focused on eosinophils in the lamina propria of the mucosa of patients with UC in the active phase. Biopsy specimens were taken from 17 patients with UC in the active phase, 17 in the inactive phase, and 20 control patients, and submitted for histochemical staining for peroxidase and chloroacetate esterase for microscopic examination. Both peroxidase-producing and chloroacetate esterase-producing cells in the lamina propria increased markedly in the active phase (8.3 ± 3.1/0.01 mm2 and 6.6 ± 2.7/0.01 mm2, respectively), compared with values in the inactive phase (0.8 ± 0.6/0.01 mm2 and 1.3 ± 0.6/0.01 mm2) or in the controls (1.3 ± 0.8/0.01 mm2 and 1.3 ± 0.4/0.01 mm2). Triple staining for peroxidase, chloroacetate esterase, and nonspecific esterase in the specimens revealed that the peroxidase-producing cells constituted a different population from that of neutrophils, macrophages/monocytes, or basophils. A monoclonal antibody specific for eosinophil peroxidase stained almost all infiltrated peroxidase-producing cells. These results indicated that eosinophils with strong peroxidase activity had infiltrated the lamina propria in UC, suggesting an allergic background and the involvement of released peroxidase in the mucosal damage characteristic of UC.

Isolation of peptides useful for differential diagnosis of Crohn’s disease and ulcerative colitis

Background: Phage displayed random peptide technology has been utilised to identify binding epitopes of antibodies or receptor ligands. Aim: To isolates peptides from a phage library which are specifically recognised by antibodies in serum from patients with Crohn’s disease (CD). Methods: A phage displayed random peptide library composed of nine amino acids was established and sequentially screened using serum immunogloblin G obtained from CD patients. Results: Five different CD specific peptides were isolated from the phage library. No homology in amino acid sequences was observed among four (CDP-1, -3 to -5) of the five peptides exhibiting different binding characteristics with each CD patient’s serum. In contrast, two peptides (CDP-1 and -2) had similar amino acid sequences and similar binding characteristics. Four multiple antigenic peptides (MAP, CDP-1, -3 to -5) were synthesised, and an enzyme linked immunosorbent assay (ELISA) using the four peptides was developed to detect serum antibodies against them. Fifty two of 92 CD patients (56.5%) were detected by ELISA, none of 20 ulcerative colitis (UC) patients, only one of 25 duodenal ulcer patients, and only three of 48 healthy subjects. Conclusions: ELISA using the four peptides isolated in this study may be useful for the differential diagnosis of CD and UC.

MUCOSAL BLOOD FLOW AND GENERATION OF SUPEROXIDE IN RAT EXPERIMENTAL COLITIS INDUCED BY SUCCINIC ACID

As we consider succinic acid to be an exacerbating factor in ulcerative colitis, we investigated its influence on rat colonic mucosa in terms of mucosal blood flow and superoxide generation. We measured mucosal blood flow by the hydrogen gas clearance method and superoxide generation by the chemiluminescence method, and observed histopathological findings to determine the effects of succinic acid. After the instillation of succinic acid of any concentration tested to the colon, mucosal blood flow decreased. Histopathologically, the higher the concentration of succinic acid, the greater was the erosion formation in the colonic mucosa, while significant polymorpho-nuclear cell infiltration and superoxide generation from colon tissue were observed with 0.01% succinic acid compared with higher or lower concentrations. Succinic acid, at fecal concentrations found in active stage ulcerative colitis, appears to be implicated in mucosal injury, mediated by a decrease in colonic mucosal blood flow and infiltration of superoxide-generating polymorpho-nuclear cells into the mucosa.

Successful eradication of Helicobacter pylori prevents relapse of peptic ulcer disease

The NIH consensus conference in 1994 recommended that all patients with peptic ulcers should be tested and treated for Helicobacter pylori. Recent studies have shown that the eradication of H. pylori is associated with a significant reduction in the relapse rate of peptic ulcers, but there are few reports about long‐term outcome.

Extracorporeal Monocyte Granulocytapheresis was Effective for a Patient of Erythema Nodosum Concomitant withUlcerative Colitis

Abstract:  We report an erythema nodosum (EN) patient whose condition becameapparent during the clinical course of ulcerative colitis (UC).The patient relapsed frequently in spite of taking a high dose adrenocorticalsteroid during his morbidity period of UC. Monocyte‐granulocytapheresis(M‐GCAP) was combined with 5‐aminosalicylic acid 2250 mg/dayperoral and once a day of steroid enema. Monocyte‐granulocytapheresiswas performed once a week for 5 weeks, and succeeded in inducingclinical remission for both UC and EN. The immunological and clinical connectionsbetween UC and EN have never been fully elucidated. In this case,because the symptoms of UC and EN revealed parallel improvementafter his inflammatory reaction had been brought under control by combining M‐GCAP therapy, we hypothesize that theonset of EN appeared as a result of the patients long‐term,treatment‐resistant immuno‐disturbance, which first appeared as symptomsof UC. Immunomodulative effects induced by M‐GCAP might help tocontrol other chronic non‐specific inflammations not concerned withtargeted organ(s).