Masanobu Matsushita

Dipeptidyl peptidase-4 inhibitor anagliptin facilitates restoration of dextran sulfate sodium-induced colitis

Abstract Objective. Inflammatory bowel disease (IBD) is a chronic debilitating disease associated with severe damage to the intestinal mucosa. Glucagon-like peptide-2 (GLP-2) is a potent and specific gastrointestinal growth factor. GLP-2 released from enteroendocrine cells is inactivated by dipeptidyl peptidase-4 (DPP-4). The aim of this study was to examine whether the DPP-4 inhibitor anagliptin improves experimental murine colitis. Material and methods. Male C57BL/6 mice aged 8 weeks were exposed to 1.5% dextran sulfate sodium (DSS) in drinking water for 7 days to induce experimental colitis. Anagliptin (0.1% in diet) was administrated from 2 days before the beginning of DSS to 7 days after the end of DSS. Changes in body weight and disease activity index were evaluated daily. Histological colitis severity, cellular proliferation and gene expression were determined in colonic tissues. Results. Treatment with anagliptin clearly improved body weight loss and disease activity index in the recovery phase. Histological score in the DSS + anagliptin group at day 14 was significantly lower than that in the DSS alone group. Treatment with anagliptin increased the Ki67-positive rate at days 10 and 14, and tended to increase insulin-like growth factor-1 mRNA expression in the DSS + anagliptin group. Conclusion. In this model of experimental colitis, the DPP-4 inhibitor anagliptin facilitated the restoration of mucosal damage, thereby resulting in the acceleration of healing. These findings suggest a new and novel therapeutic approach for the treatment of IBD.

Importance of appropriate pharmaceutical management in pregnant women with ulcerative colitis

BackgroundUlcerative colitis (UC) often occurs in women of childbearing age. Compared to Western countries, however, few studies have investigated the impact of UC on the progress of pregnancy in Asian populations.MethodsWe retrospectively examined 91 pregnancies in 64 patients with UC experienced at our hospital and related institutions from 1991 to 2011, focusing on the relationship between the progression of UC during pregnancy, progress of the pregnancy itself, and the treatment of UC.ResultsIn 80 of 91 pregnancies the patient had already been diagnosed with UC at the time she became pregnant, of whom 31 (38.8%) experienced exacerbation during pregnancy. Regarding severity, moderate or severe active-stage disease during pregnancy was seen in 13.7% of those who had been in remission at the onset of pregnancy versus 58.6% of those who had been in the active stage at onset (OR 8.9: 95%CI 3.0~26.4; P<0.01). The incidence of miscarriage or abortion was 9.8% in pregnancies in which UC was in remission at onset versus 31% in those in which it was in the active stage at onset (OR 4.1: 95%CI 1.2~13.9; P=0.02). Among patients, 62.5% were receiving pharmaceutical treatment at onset of pregnancy. Exacerbation during pregnancy occurred in 26.5% of the group who continued to receive the same treatment during pregnancy versus 56.3% of those with a dose decrease or discontinuation after onset (OR 3.6: 95%CI 1.0~12.4; P=0.04).ConclusionsUC patients wishing to conceive should do so when in remission and continue appropriate pharmaceutical treatment during pregnancy.

Usefulness of Helicobacter pylori eradication for precancerous lesions of the gastric remnant.

Secondary stomach cancer in lesions of the remnant stomach occurs relatively soon after distal gastrectomy using the Billroth I reconstruction procedure. Prophylactic eradication of Helicobacter pylori after endoscopic resection of early gastric cancer should be used to prevent the development of metachronous gastric carcinoma. However, the effect of H. pylori eradication on the gastric remnant has not been clearly determined.

Nomogram-based prediction of rebleeding in small bowel bleeding patients: the ‘PRSBB’ score

Small bowel capsule endoscopy (SBCE) and balloon-assisted endoscopy (BAE) have revolutionized the diagnosis and treatment of small bowel bleeding (SBB), allowing access to the small bowel and identification of specific bleeding lesions. However, some patients experience rebleeding after small bowel investigation, and there are no definitive algorithms for determining the most appropriate follow-up strategy in SBB patients. We developed and validated a nomogram that can predict rebleeding risk and be used to develop a risk-stratified follow-up strategy in SBB patients. A retrospective study was performed using data from 401 SBB patients who underwent SBCE at Nagoya University Hospital. We developed and internally validated a predictive model for rebleeding in the form of a nomogram using Cox regression models and a bootstrap resampling procedure. Optimal risk factors were selected according to the least absolute shrinkage and selection operator (LASSO). The LASSO method identified 8 independent predictors of rebleeding that could be assessed to obtain a ‘predicting rebleeding in SBB’, or ‘PRSBB’ score: age, sex, SBB type, transfusion requirement, cardiovascular disease, liver cirrhosis, SBCE findings, and treatment. The c-statistic for the predictive model was 0.681. In conclusion, our PRSBB score can help clinicians devise appropriate follow-up plans.

In vivo histological diagnosis for gastric cancer using endocytoscopy

AIM To examine usefulness of virtual biopsy using endocytoscopy by comparing the in vivo endocytoscopic and histopathological images of gastric cancers. METHODS Endocytoscopy was performed in 30 patients with early gastric cancer. Of these, 26 patients showed well differentiated adenocarcinomas, while 4 patients showed poorly differentiated adenocarcinomas (including one signet ring cell carcinoma). Cancerous and non-cancerous areas were observed after double staining with 0.05% crystal violet and 0.1% methylene blue. The endocytoscopic images obtained were evaluated by an expert endoscopist and an expert pathologist without knowledge of patient clinical data, and endocytoscopic and histopathological diagnoses were compared. RESULTS The endocytoscopic images of the cancerous area were assessed as evaluable in 25 (83.3%) and 27 (90%) patients by endoscopist A and pathologist B, respectively, and those of the non-cancerous area as evaluable in 28 (93.3%) and 23 (76.7%) patients by the endoscopist and pathologist, respectively. The sensitivity, specificity, and diagnostic accuracy of gastric cancer diagnosis using evaluable endocytoscopic images were 88.0% and 92.9%, and 90.6% by endoscopist A, and 88.9% and 91.3%, and 90.0% by pathologist B, respectively. Evaluation of the diagnostic concordance rate between the endoscopist and the pathologist by inter-observer agreement calculation revealed no significant difference between the two observers. The inter-observer agreement (κ-value) for endocytoscopic diagnosis was 0.745. CONCLUSION Endocytoscopy is useful for the differentiation of cancerous from non-cancerous gastric mucosa, making it a promising tool for virtual biopsy.

Inhibition of KDM4A activity as a strategy to suppress interleukin-6 production and attenuate colitis induction.

4-Chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) functions as a hapten and fluoresces upon binding to proteins. Therefore, fluorescence visualization of hapten-proteins is a feature of the colitis induced by NBD-Cl. Using this colitis model, we located activated fibroblasts in the vicinity of hapten-proteins upon colitis induction and observed interleukin (IL)-6 production in the activated fibroblasts. We screened herbal ingredients using primary fibroblasts stimulated with tumor necrosis factor α (TNF-α) and found the suppressive action of Atractylodin on IL-6 production. Under TNF-α stimulation, Atractylodin induced the tri-methylation of histone H3 at lysine residue 9, which impaired the binding between NF-κB and the IL-6 promoter on the genomic DNA. Atractylodin inhibited KDM4A but not KDM6A activity. Atractylodin administration attenuated colitis induction. The KDM4A inhibitor ML324 showed similar actions on IL-6 production and colitis induction. We propose the inhibition of KDM4A activity as a strategy to suppress IL-6 production and attenuate colitis induction.

Combinational use of lipid-based reagents for efficient transfection of primary fibroblasts and hepatoblasts

Commercially available lipid-based transfection reagents are widely used to deliver DNA to cells. However, these lipid-based transfection reagents show poor gene transfer efficiency in primary cells. Here, we demonstrate a simple method to improve gene transfer efficiency in primary fibroblasts and hepatoblasts using a combination of lipid-based transfection reagents. Our data show that combined use of Lipofectamine LTX and FuGENE HD increases the efficiency of gene transfer compared with the use of either reagent alone, and this combination achieves the best result of any pairwise combination of Lipofectamine LTX, FuGENE HD, TransFectin, and Fibroblast Transfection Reagent.

Small intestine morphology and recovery after drug-induced colitis in proglucagon-derived peptides knockout mice

Abstract Objective. Dipeptidyl peptidase (DPP)-4 inhibitors inactivate glucagon-like peptide (GLP)-1, GLP-2, and other hormones by degradation. Administration of a DPP-4 inhibitor or GLP-2 analog stimulates intestinal growth and facilitates the restoration of mucosal damage caused by drug-induced colitis in mice. We studied the effect of GLP-2 deletion on morphological changes on gut and colitis recovery in mice deficient in proglucagon-derived peptides (PGDPs). Material and methods. Mice deficient in PGDPs were used. Eight-week-old male PGDPs−/− and PGDPs+/+ mice were sacrificed to examine the small intestine and colon morphology. To elucidate the effect of PGDP deletion on colitis, 12-week-old PGDPs−/− and PGDPs+/+ male mice were exposed to 2.5% dextran sulfate sodium (DSS) for 6 days. Colitis severity was assessed daily by disease activity index and body weight loss. Histological examinations were performed on days 7 and 21. Results. There were no differences in the height, width, and density of villous and the depth of crypt of small intestine, and the weight, length, and the crypt depth of colon between PGDPs−/− and PGDPs+/+. In DSS colitis, the decline and amelioration of disease activity index and body weight were not different between the PGDP−/− and PGDP+/+ mice. Histologically, the PGDP−/− mice showed significantly less recovery than showed by the PGDPs+/+ mice on day 21 (p < 0.05). Conclusion. Mice with PGDP deletion did not show morphological changes of gut. On colitis, PGDP deletion only caused less histological amelioration of colitis in the later phase of recovery.

Su1241 Short- and Long-Term Efficacy of Tacrolimus in Patients With Refractory Ulcerative Colitis

disease (CD) or ulcerative colitis (UC)], had a measured low vit D 25 OH level ( ,30 ng/ mL), and were treated with high-dose vit D received high-dose vit D treatment (ergocalciferol 50,000 units once a week for eight weeks). Outcomes included duel-energy x-ray absorptiometry (DEXA) scan measures and repeat vit D 25 -OH level if obtained before and after high dose vit D treatment. Statistical analysis included Wilcoxon signed-rank test. Results: 52 pts with IBD (39 CD, 13 UC), low vit D 25 OH levels, and treatment with high dose vit D were identified. 34 were male (66%), and the median age was 57 (range 22, 83 yrs). 14 (27%) had one or more IBD-related surgeries. 11 were smokers (22%). 7 (13%) pts were on corticosteroids, 8 (15%) pts were on budesonide, 20 (38%) were on immunomodulator therapy, and 20 (38%) were on anti-TNF medications. 19 (37%) pts had a previous diagnosis of osteopenia, and 12 (23%) had osteoporosis. 19 (37%) were taking a bisphosphonate. 24 (46%) had documented daily non-prescription vit D supplementation. 29 pts had a DEXA scan before and after high dose vit D treatment after a median of 2.1 yrs (range 1, 6.5 yrs). Spine bone mineral density (BMD) scores improved significantly (1.05 ± 0.17 to 1.10 ± 0.20, p , 0.05). There was no significant change in BMD of the hip (0.79 ± .15 to 0.75 ± .31), and hip and spine T scores and Z scores did not change significantly. 36 pts had repeat vit D 25 OH levels checked after a median of 120 days (range 47, 964 days). Vit D 25 OH increased significantly (15.5 ± 8.4 to 29.4 ± 19.0, p , 0.05). However, 14 (36%) pts remained vit D deficient (vit D 25 OH levels , 20 ng/mL), and 8 (22%) pts remained vit D insufficient (vit D 25 OH levels 20-30 ng/mL). Persistent vit D insufficiency was not dependent on gender, surgical history, medication type, or disease type. Conclusion: In this study, the majority patients with IBD who are vit D insufficient failed to achieve normal serum vit D levels after treatment with high dose vit D, but bone health based on DEXA scan did not worsen significantly. Further research into appropriate calcium and vit D dosing in this patient population is warranted.