Severe Mucosal Damage Research Articles

Objective evaluation of nutritional status using the prognostic nutritional index during and after chemoradiotherapy in Japanese patients with head and neck cancer: a retrospective study

ObjectivesThe incidence of severe mucosal damage due to low nutritional status is high in patients receiving concurrent chemoradiotherapy (CCRT) for head and neck cancer. Objective assessments do not exist for...

Loss of C/EBPδ enhances apoptosis of intestinal epithelial cells and exacerbates experimental colitis in mice.

Inflammatory bowel diseases (IBDs) are characterized by chronic inflammation involving intestinal tissue damage, which include ulcerative colitis and Crohn's disease as major entities. Accumulating evidence suggests that excessive apoptosis of intestinal epithelial cells (IECs) contributes to the development of IBD. It was recently reported that the transcription factor CCAAT/enhancer-binding protein delta (C/EBPδ) is involved in inflammation; however, its role in colitis remains unclear. Here, we found that C/EBPδ knockout mice showed enhanced susceptibility to dextran sodium sulfate (DSS)-induced colitis, a mouse model of IBD, which was associated with severe colonic inflammation and mucosal damage with increased IEC apoptosis. Additionally, DSS stimulation induced increased expression of pro-apoptotic BH3-only protein Bim in the colon of C/EBPδ knockout mice. Collectively, our findings demonstrate that C/EBPδ plays an essential role in suppressing DSS-induced colitis, likely by attenuating IEC apoptosis.

Effects of Western Diet on Giardiasis: A Role for Fatty Acids and Gut Microbiota in the Persistence and Severity of Giardia Infections

AIMSGiardia duodenalis is a protozoan parasite responsible for giardiasis, one of the most common cause of waterborne diseases. Giardia. sp causes epithelial disruptions as well as microbiota alterations. Giardia infections are common in countries with poor sanitation, where the diet is mostly agrarian and high in fiber content. Intriguingly, in such environments, giardiasis has commonly been reported to cause little or no illness. Giardiasis also occurs in developed countries with western/high fat diets, where infections are a common cause of diarrhea. This sutdy investigated the role of diet on the severity and the duration of giardiasis. We hypothesized that western diet may contribute to the persistence of Giardia in the gut. Experiments assessed the role of fatty acids and gut microbiota in pathophysiology.METHODSC57/BL6 mice were fed for two weeks with 5TJN Western diet or 5TJS Low‐Fat (LF) diet prior to infection with Giardia duodenalis GS/M strain (human isolate). The small intestines were collected at day 7 post infection (PI) for histological study and parasite burden. Gut microbiota composition was determined by Illumina Mi‐Seq 16S RNA sequencing. The effect of Oleic and Palmitic acid on Giardia metabolic activity was determined by resazurin assay. Finally, tight junction disruption of small intestinal epithelial cell line SCNB infected with G. duodenalis Assemblage A (NF strain) and Assemblage B (GS/M strain) in the presence of Oleic, Palmitic acid (0.3 mM) or both was determined by immunofluorescence.RESULTSWe observed that Giardia infected C57/BL6 mice that were fed with 5TJN Western diet exhibited a significantly higher trophozoite burden 7 days PI. Giardia infected mice also exhibited more severe mucosal damage (lower villus‐crypt ratio) when fed with Western diet. Fecal microbiota analysis showed that mice fed with Western Diet exhibit a higher abundance of Verrucomicrobia and a lower abundance of Firmicutes than mice fed LF diet. Upon Giardia infection, a decrease of Bacteroidetes and an increase of Firmicutes was observed. Overall, Giardia infection induced an increase of the alpha diversity in Western mice compared to mice given LF diet. Trophozoite metabolic activity was enhanced in the presence of Oleic acid (0.1 mM) and Palmitic acid (0.25mM). In addition, small intestine cell lines infected with G. duodenalis strain NF and GS/M showed a disruption and/or a rearrangement of tight junction proteins such as ZO‐1, Claudin‐1 and 4 when incubated with Oleic and Palmitic acids (0.3 mM).CONCLUSIONThis study gives insight into how diet may influence the outcome of parasitic diseases and intestinal pathophysiology. The findings suggest that a Western Diet may increase the severity of giardiasis by, at least in part, promoting the growth of Giardia through fatty acid supplementation and microbiota modulation. The potential of diet intervention in treatment against parasitic diseases warrants further research.Support or Funding InformationGrant support: NSERCThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Schistosoma mansoni Coinfection Attenuates Murine Toxoplasma gondii-Induced Crohn's-Like Ileitis by Preserving the Epithelial Barrier and Downregulating the Inflammatory Response.

Background and aims: Mice orally infected with T. gondii develop Crohn's disease (CD)-like enteritis associated with severe mucosal damage and a systemic inflammatory response, resulting in high morbidity and mortality. Previously, helminthic infections have shown therapeutic potential in experimental colitis. However, the role of S. mansoni in T. gondii-induced CD-like enteritis has not been elucidated. Our study investigated the mechanisms underlying T. gondii-induced ileitis and the potential therapeutic effect of S. mansoni coinfection.Methods: C57BL/6 mice were infected by subcutaneous injection of cercariae of the BH strain of S. mansoni, and 7–9 weeks later, they were orally infected with cysts of the ME49 strain of T. gondii. After euthanasia, the ileum was removed for histopathological analysis; staining for goblet cells; immunohistochemistry characterizing mononuclear cells, lysozyme expression, apoptotic cells, and intracellular pathway activation; and measuring gene expression levels by real-time PCR. Cytokine concentrations were measured in the serial serum samples and culture supernatants of the ileal explants, in addition to myeloperoxidase (MPO) activity.Results: T. gondii-monoinfected mice presented dense inflammatory cell infiltrates and ulcerations in the terminal ileum, with abundant cell extrusion, apoptotic bodies, and necrosis; these effects were absent in S. mansoni-infected or coinfected animals. Coinfection preserved goblet cells and Paneth cells, remarkably depleted in T. gondii-infected mice. Densities of CD4- and CD11b-positive cells were increased in T. gondii- compared to S. mansoni-infected mice and controls. MPO was significantly increased among T. gondii-mice, while attenuated in coinfected animals. In T. gondii-infected mice, the culture supernatants of the explants showed increased concentrations of TNF-alpha, IFN-gamma, and IL-17, and the ileal tissue revealed increased expression of the mRNA transcripts for IL-1 beta, NOS2, HMOX1, MMP3, and MMP9 and activation of NF-kappa B and p38 MAPK signaling, all of which were counterregulated by S. mansoni coinfection.Conclusion: S. mansoni coinfection attenuates T. gondii-induced ileitis by preserving mucosal integrity and downregulating the local inflammatory response based on the activation of NF-kappa B and MAPK. The protective function of prior S. mansoni infection suggests the involvement of innate immune mechanisms and supports a conceptually new approach to the treatment of chronic inflammatory diseases, including CD.

Intestinal anti-inflammatory activity of xique-xique (Pilosocereus gounellei A. Weber ex K. Schum. Bly. Ex Rowl) juice on acetic acid-induced colitis in rats.

Inflammatory bowel disease (IBD) is characterized by severe mucosal damage in the intestine and a deregulated immune response. Natural products derived from plants that are rich in bioactive compounds are used by many patients with IBD. Xique-xique (Pilosocereus gounellei) is a cactus of the Caatinga family that has been used by the local population for food and medicinal purposes. The intestinal anti-inflammatory effect of xique-xique cladode juice was evaluated in the present study. A dose of 5 mL kg-1 had a protective effect on intestinal inflammation, with an improvement in macroscopic damage, and a decrease in pro-inflammatory markers and oxidative stress, in addition to preserving the colonic tissue. Immunohistochemical analysis revealed the downregulation of IL-17, NF-κB, and iNOS, and upregulation of SOCs-1, ZO-1, and MUC-2. These protective effects could be attributed to the phenolic compounds as well as the fibers present in xique-xique juice. Further studies are needed before suggesting the use of xique-xique juice as a new alternative for treating IBD.

Rosuvastatin attenuates piroxicam-mediated gastric ulceration and hepato-renal toxicity in rats

Piroxicam (Px) is a non-steroidal anti-inflammatory drug that is widely prescribed in various inflammatory disorders. However, Px is known to induce gastric ulceration and hepato-renal toxicity. Rosuvastatin (ROSV), a member of the statin family, has anti-inflammatory and antioxidant actions independent of its anti-hyperlipidemic action. Therefore, we investigated the protective effects of ROSV against Px-induced gastric, liver, and kidney injury. Five groups of seven rats each were used; control group, ROSV group (20 mg/kg, given orally), Px group (7 mg/kg, given intraperitoneally), Px+ROSV L (7 and 10 mg/kg, respectively), and Px+ROSV H (7 and 20 mg/kg, respectively) group. The results revealed that Px induced severe gastric mucosal damage expressed by high ulcer index along with significant increases in liver and kidney function parameters including AST, ALT, creatinine, and urea. Disrupted lipid metabolism also was observed in Px-treated animals. Moreover, marked an increase in malondialdehyde (MDA) and decreases in glutathione (GSH) and catalase (CAT) levels along with enhanced activated caspase-3 expression in the gastric, hepatic, and renal tissues following Px-insult suggested a possible involvement of lipid peroxidation in Px-induced gastropathy and hepatorenal toxicity. However, in a dose-dependent manner, ROSV was able to mitigate Px-induced lipid peroxidation and apoptosis in gastric, liver, and kidney tissues.

Outcome of gastrointestinal graft-versus-host disease according to the treatment response

The optimal treatment strategy for gastrointestinal graft-versus-host disease (GI-GVHD) after allogeneic hematopoietic cell transplantation remains to be established. We retrospectively analyzed 68 cases of GI-GVHD at our institution between 2007 and 2017. The survival outcomes were significantly inferior in patients who did not respond to the first-line treatment (1-year overall survival 27.3 vs 69.2%, P = 0.0017; non-relapse mortality 50.0 vs 18.6%, P = 0.026). After subsequent treatments, 18 patients were refractory to all steroid-based treatments such as steroid pulse therapy and oral beclomethasone dipropionate (BDP). However, these steroid-refractory cases showed a gradual increase in the response rate after the initial diagnosis of steroid refractoriness. This result may be explained by the problem of evaluating the response based solely on the volume of diarrhea, i.e., severe mucosal damage due to refractory GI-GVHD may require a long recovery and sometimes be complicated with other diseases. In conclusion, patients with GI-GVHD who failed to respond to the first-line treatment had inferior survival. However, later improvement may be observed without additional immunosuppressant other than steroid among patients who initially do not respond to steroid therapy. It is important to repeat colonoscopy in patients with refractory GI-GVHD to monitor the activity of GVHD.

Functional characterization of oxazolone-induced colitis and survival improvement by vagus nerve stimulation.

BackgroundOxazolone-induced colitis has been frequently used in literature as a model of IBD, but insights into the underlying immune response and pathological features are surprisingly still very limited. Vagus nerve stimulation (VNS) has proven to be effective in innate and Th1/Th17 predominant inflammatory models, including pre-clinical models of colitis, however to what extent VNS is also effective in ameliorating Th2-driven colitis remains to be studied. In the present study, we therefore further characterized the immune response in oxazolone-induced colitis and investigated the potential therapeutic effect of VNS.MethodsColitis was induced in Balb/c mice by cutaneous sensitization with 3% oxazolone followed by intracolonic administration of 1% oxazolone 7 days later. To evaluate the effect of VNS on the development of Th2-driven colitis, VNS and sham-treated mice were challenged with 1% oxazolone.ResultsIntracolonic oxazolone administration resulted in a severe destruction of the colonic mucosa and a rapid drop in body temperature leading to a 65% mortality rate at day 5. Severe infiltration of neutrophils and monocytes was detected 6h after oxazolone administration which was associated with a Th2-type inflammatory response. VNS significantly improved survival rate which correlated with decreased levels of HMGB1 and reduced colonic (il6 and cxcl1 mRNA) and serum cytokine levels (IL-6, TNFα and CXCL1) compared to sham treated mice.ConclusionsOxazolone-induced colitis rather represents a model of sepsis and, at best, may resemble a severe type of ulcerative colitis, associated with early and severe mucosal damage and a high mortality rate. VNS reduces colonic inflammation and improves survival in this model, supporting the anti-inflammatory properties of VNS, even in an aggressive model as oxazolone-induced colitis.

Contributions of HLA haplotypes, IL8 level and Toxoplasma gondii infection in defining celiac disease's phenotypes

BackgroundIt is not clear why some patients with coeliac disease (CD) present with severe symptoms and small intestinal mucosal damages while others present with milder symptoms and no frank enteropathy. There is no study to assess the associated factors with mild/severe symptoms and enteropathy. The terminologies like latent, silent and potential are difficult to use and are unrepresentative. In the present study we describe coeliac disease’s phenotypes based on HLA haplotypes, IL8 production and past infection with Toxoplasma gondii (T. gondii) infection.MethodsIn this case-control study, sera originating from 150 healthy subjects and 150 patients diagnosed with CD during the years 2013–14 were analyzed for the presence of antibodies specific T. gondii of the IgG and IgM subclasses. The level of IL8 were measured and HLA-DQ2 and HLA-DQ8 alleles were genotyped. The correlation between these parameters and the damages in intestinal mucosal were assessed using an accepted histopathological classification.ResultsHigh levels of IgG antibodies against T. gondii were found in the sera of control group compared to the CD group (52.6% vs. 39.4%, P = 0.02). Mean serum levels of IL8 was significantly higher in CD patients compared with control (P ≤ 0.05). By comparing the level of anti- T. gondii IgG and mucosal damage in celiac disease, we found a significant relationship between the severity of mucosal damages and anti- T. gondii IgG level (P = 0.02). No correlation was detected between Toxoplasma gondii infection and types of HLA (P > 0.05). However, patients with severely abnormal histology carried HLA-DQ2 risk alleles (92 patients (61%)) more often than the controls and those with mild histological abnormalities.ConclusionsCD patients with severe histological changes had more often Toxoplasma gondii infection than those affected with mild histological features. This suggests that CD’s phenotypes are correlated to additional factors like infections and to particular HLA DQ2 alleles that may need additional investigations and potentially will require additional treatment.

239 - Bowel radiation injury: Promises of cell and tissue engineering

Bowel radiation injury is an insidious disease associated with substantial morbidity and mortality. Moreover, it's an increasing problem as more patients receive radiotherapy and survive longer after their tumor treatment. Bowel radiation injury results from the treatment of several cancers by radiotherapy in which normal colorectal tissues are present in the irradiation field. The clinical expression of bowel complications associated to radiotherapy resembles chronic bowel disease of other etiologies. However, recent studies have identified differences and specialists have proposed that complications following pelvic radiotherapy should be recognized as a “new disease” by Andreyev et al. The growing number of cases declared every year highlights the importance of understanding the mechanisms involved and of finding effective therapies. There is no unified approach for the assessment and treatment of this disease partly due to insufficient knowledge about the mechanism involved in the development of bowel radiation injury. However, unresolved inflammation is hypothesized to have an important role in late side effects. We used an experimental model of radiation proctitis developed in rats that reproduces severe colonic mucosal damages and fibrosis similar to those observed in patients treated by radiotherapy (Sémont, 2010). We evaluated the benefit of immunomodulatory mesenchymal stromal cells isolated from adipose tissue (Ad-MSC) to reduce late side effects (Bessout, 2013–15 and Durand, 2015). We demonstrated a therapeutic benefit on different crucial functions of the colon and determined pleiotropic action mechanisms of the cell therapy treatment. Since, MSC therapy has been proposed to 4 patients over-irradiated during their radiotherapy treatment leading to substantial improvement in the clinical response (Voswinkel, 2013). Our pre-clinical studies also identified targets to potentiate the therapeutic effect of Ad-MSC. We have thus highlighted a biomaterial-assisted MSC therapy to alleviate colonic radiation-induced damage (Moussa, 2017). Moreover, we also tested a new treatment procedure to optimize the paracrine effect of MSC.

Effects of the RNA-binding protein, KSRP, on innate immune response against Helicobacter pylori infection in mice

Helicobacter pylori (H. pylori) contributes to various gastric diseases such as chronic gastritis, gastric ulcer, and gastric carcinoma. Host innate immune response against the pathogen plays a significant role in elimination of pathogen infection. Importantly, pathogen elimination is closely related to numerous inflammatory-related genes that participate in complex biological response of cells to harmful stimuli. Here we studied effects of the KH-type splicing regulatory protein (KSRP), a RNA-binding protein, on innate immune response against H. pylori infection. We found that H. pylori infection downregulated KSRP expression directly, and that KSRP overexpression repressed upregulation of CXCL-2 expression induced by H. pylori and facilitated H. pylori proliferation in vitro. Similarly, KSRP overexpression in H. pylori mice also facilitated H. pylori proliferation and colonization, and induced more severe gastric mucosal damage. Intriguingly, CXCL-2 and HMOX-1 were upregulated in H. pylori infected mice after KSRP overexpression. This difference in expression of these genes implicated that KSRP was closely associated with and directly participated in the innate immune response against H. pylori. These results were beneficial for understanding the in vivo function of KSRP on innate immune response against pathogen infection.

Autophagy can alleviate severe burn-induced damage to the intestinal tract in mice

The present study was designed to examine the effect of autophagy and apoptosis on intestinal injury in mice after severe burns. Kunming mice were subjected to third degree burns over 30% of the total body surface area. Damage to the intestine was assessed by examining changes in intestinal mucosal morphology, enzyme-linked immunosorbent assay of serum d-lactate, diamine oxidase, lipopolysaccharide, interleukin-6, and tumor necrosis factor α (marker of intestinal damage), hematoxylin and eosin staining, and Western blotting under 4 experimental conditions: control group, burn only (burn group), burn and administration of rapamycin to stimulate intestinal autophagy (rapamycin group), or burn and administration of 3-methyladenine to inhibit intestinal autophagy (3-methyladenine group). At day 1 postburn, the expression levels of light chain 3 II, beclin-1, and cleaved caspase-3 were significantly greater in all 3 groups of mice subjected to the burn injury than in the control group 1day postburn; while the levels of light chain 3 II and beclin-1 were significantly greater and those of cleaved caspase-3 were significantly less in the rapamycin group than in the burn group. In contrast, light chain 3 II and beclin-1 levels were significantly less and those of cleaved caspase-3 significantly greater in the 3-methyladenine group. All 3 groups subjected to burn injury showed significantly increased levels of d-lactate, diamine oxidase, lipopolysaccharide, interleukin-6, and tumor necrosis factor α. Of the 3 groups, the rapamycin group exhibited the least observed levels, the 3-methyladenine group the greatest, and the burn group intermediate. Pathologic sections of the intestinal tissue showed that all 3 burn groups exhibited severe intestinal mucosal damage at 1day postburn. The condition of the 3-methyladenine treatment group was worse than that of the rapamycin treatment group, but better than that of the burn group. Intestinal autophagy is activated in response to intestinal apoptosis after severe burns and may alleviate burn-induced intestinal injury.

Dextran sulfate sodium-induced colitis mice up-regulated extracellular matrix tenascin-C

Tenascin-C, an extracellular matrix glycoprotein, expresses high level during embryogenesis and almost absent during the normal postnatal life. However, it is re-appeared in a diverse condition such as tissue injury and in the stroma of various carcinomas. In this study, we investigated the appearance of tenascin-C in dextran sulfate sodium (DSS)-induced colitis in mice. DSS induced colitis mice demonstrated severe mucosal damage, with distortion and loss of crypts, depletion of goblet cells and infiltration of macrophages particularly F4/80 positive macrophages, granulocytes and lymphocytes in the colon tissues. These DSS inflamed colon expressed a high and dense level of tenascin-C in the severe damaged areas, whereas, moderate staining was observed in the moderate inflamed areas. DSS-induced colitis mice significantly increased macrophages infiltration in the colon tissues. These results suggested that tenascin-C extracellular matrix re-appeared in the colon tissues during inflammation.Asian J. Med. Biol. Res. December 2016, 2(4): 582-586

Clinical Evaluation of "Shock Bowel" Using Intestinal Fatty Acid Binding Protein.

"Shock bowel" is one of the computed tomographic (CT) signs of hypotension, yet its clinical implications remain poorly understood. We evaluated how shock bowel affects clinical outcomes and the extent of intestinal epithelial damage in trauma patients by measuring the level of intestinal fatty acid binding protein (I-FABP). We reviewed the initial CT scans, taken in the emergency room, of 92 patients with severe blunt torso trauma who were consecutively admitted during a 24-month period. The data collected included CT signs of hypotension, I-FABP, feeding intolerance, and other clinical outcomes. Demographic and clinical outcomes were compared in patients with and without hemodynamic shock and shock bowel. Shock bowel was found in 16 patients (17.4%); of them 7 patients (43.8%) did not have hemodynamic shock. Certain CT signs of hypotension, namely free peritoneal fluid, contrast extravasation, small-caliber aorta, and shock bowel, were significantly more common in patients with hemodynamic shock than in patients without (P < 0.05). Injury severity score and the rate of consciousness disturbance were significantly higher in patients with shock bowel than in patients without (P < 0.05). The rate of feeding intolerance and median plasma I-FABP levels were significantly higher in patients with shock bowel than in patients without (75.0% vs. 22.4%, P < 0.001 and 17.0 ng/mL vs. 3.7 ng/mL, P < 0.001, respectively). There was no difference in mortality. In conclusion, shock bowel is not always due to hemodynamic shock. It does, however, indicate severe intestinal mucosal damages and may predict feeding intolerance.

Unique Cellular Lineage Composition of the First Gland of the Mouse Gastric Corpus.

The glandular stomach has two major zones: the acid secreting corpus and the gastrin cell-containing antrum. Nevertheless, a single gland lies at the transition between the forestomach and corpus in the mouse stomach. We have sought to define the lineages that make up this gland unit at the squamocolumnar junction. The first gland in mice showed a notable absence of characteristic corpus lineages, including parietal cells and chief cells. In contrast, the gland showed strong staining of Griffonia simplicifolia-II (GSII)-lectin-positive mucous cells at the bases of glands, which were also positive for CD44 variant 9 and Clusterin. Prominent numbers of doublecortin-like kinase 1 (DCLK1) positive tuft cells were present in the first gland. The first gland contained Lgr5-expressing putative progenitor cells, and a large proportion of the cells were positive for Sox2. The cells of the first gland stained strongly for MUC4 and EpCAM, but both were absent in the normal corpus mucosa. The present studies indicate that the first gland in the corpus represents a unique anatomic entity. The presence of a concentration of progenitor cells and sensory tuft cells in this gland suggests that it may represent a source of reserve reparative cells for adapting to severe mucosal damage.

Comparison of Rate and Severity of Nasal Mucosal Tear with Internal and External Lateral Nasal Osteotomy Using Endoscope

1. Abstract 1.1.Background: A precise and reproducible lateral Osteotomy is a necessity for successful rhinoplasty. Generally, two basic techniques have been introduced: the external perforated method and the internal continuous method. There is an ongoing debate in the literature on the appropriateness of each method in terms of post-operative complications and patients’ satisfaction. Nasal mucosal tear is a key element in this comparison. This study was designed to compare the two techniques using a blinded endoscopic evaluation of the nasal mucosa tear after the osteotomies were performed by one of these two techniques. 1.2.Methods & materials: In a randomized clinical trial, 30 candidates for elective nasal surgery (aged 20-30 years) were studied in Tabriz Imam Reza Teaching Center during a 12-month period. Each patient underwent an external lateral Osteotomy performed on one side and an internal lateral osteotomy performed on the alternate side by a specialist with experience in the use of both types of osteotomy. In a blinded manner, another investigator used nasal endoscopy to detect nasal mucosal tears and reported them in millimeter (mm). 1.3.Results: A total of 30 patients, 10 males and 20 females with the mean age of 26.47±7.36 years (20-30) were enrolled in this study. Nasal mucosal tear was reported in all patients with internal lateral osteotomy, whereas this rate was 90% in the other group (p=0.24, odds ratio=1.11, 95% confidence interval=0.99-1.25). The median length of the mucosal tears was 9mm (Interquartile range, IQR=2) in the cases with internal osteotomy vs. 4mm (IQR=1) in the patients with external approach. The difference was statistically significant (p<0.001). The intra-group analysis stratified by the patients age and sex revealed a significant association neither with frequency nor with severity of the nasal mucosal tears. Conclusion: This study showed that internal osteotomy, as compared with external technique, is along with significantly more severe nasal mucosal damage in patients undergoing elective nasal surgery.

Gastroprotective effects of Kangfuxin-against ethanol-induced gastric ulcer via attenuating oxidative stress and ER stress in mice

Oxidative stress and ER stress play a role in the pathogenesis of gastric ulcer. Kangfuxin (KFX) has been used to treat gastric ulcer in patients. However, the underlying mechanisms of KFX action remain unclear. The current study was undertaken to evaluate the gastroprotective effects of KFX and to determine its potential mechanisms. Ethanol-induced gastric ulcer mouse model was employed. Ethanol pretreated mice were treated with low (0.02 g/kg) and high (0.05 g/kg) dose of KFX for 14 days. Cimetidine (0.8 g/kg) was used as positive control. Histological evaluation of the gastric mucosa revealed that mice treated with ethanol exhibited severe gastric mucosal damage. Ethanol treatment increased plasma and gastric MDA level, decreased plasma and gastric SOD activity, and reduced gastric HO-1 and GCL-c mRNA levels. ER stress markers (CHOP, GRP78, and caspase 12) were up-regulated upon ethanol administration. Moreover, increased cell apoptosis and pro-apoptotic protein Bax and caspase 3 were observed in ethanol treated mice, while the anti-apoptotic protein Bcl 2 was inhibited. Finally, KFX treatment reversed ethanol-induced phenotypes and ameliorated gastric ulcer. Our results demonstrated that the gastroprotective effects of KFX against ethanol-induced gastric ulcer could be attributed to its anti-oxidative stress, anti-ER stress and anti-apoptotic effects.

Role of proneurotensin as marker of paediatric coeliac disease.

Neurotensin (NT) is a gut hormone functioning proinflammatory through nuclear factor kappa B (NF-κB) and interleukin (IL)-8 secretion or anti-inflammatory through epidermal growth factor receptors. NT mRNA is down-regulated in duodenal biopsies of children with untreated coeliac disease. The aim of this study was to investigate if plasma pro-NT levels correlated with the degree of intestinal mucosal damage and tissue transglutaminase autoantibody (tTGA) levels in children with coeliac disease. Fasting plasma samples from 96 children with coeliac disease and 89 non-coeliac disease controls were analysed for NT precursor fragment pro-NT 1-117 by a chemiluminometric immunoassay. Pro-NT levels were compared with NT mRNA from duodenal biopsies, assessed previously with quantitative polymerase chain reaction (PCR). Illumina core exome arrays were used for human leucocyte antigen (HLA) typing and the Marsh criteria applied to score mucosal damage. Tissue TGA was measured by radio binding assay. A general linear model compared pro-NT levels with diagnosis of coeliac disease, Marsh score and HLA DQ haplotype. Spearman's rank test was used to compare pro-NT levels with tTGA, age and duodenal NT mRNA levels, respectively. Plasma pro-NT levels were elevated in children with coeliac disease (median 23 pmol/l higher, P = 0·003) and in those with severe intestinal mucosal damage (median 24 pmol/l higher for ≥ Marsh 3b versus not, P = 0·0004). Pro-NT levels correlated further with tTGA (r2 = 0·22, P = 0·002), but not with duodenal NTS mRNA levels (r2 = -0·12, P = 0·14). Pro-NT was not associated with any of the HLA risk-haplotypes. Elevated peripheral pro-NT levels reflect more severe forms of active coeliac disease, indicating a potential role of NT in intestinal inflammation.

Selective scavenging of intra-mitochondrial superoxide corrects diclofenac-induced mitochondrial dysfunction and gastric injury: A novel gastroprotective mechanism independent of gastric acid suppression

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat multiple inflammatory diseases and pain but severe gastric mucosal damage is the worst outcome of NSAID-therapy. Here we report that mitoTEMPO, a mitochondrially targeted superoxide (O2−) scavenger protected as well as healed gastric injury induced by diclofenac (DCF), the most commonly used NSAID. Common existing therapy against gastric injury involves suppression of gastric acid secretion by proton pump inhibitors and histamine H2 receptor antagonists; however, dyspepsia, vitamin B12 deficiency and gastric microfloral dysbalance are the major drawbacks of acid suppression. Interestingly, mitoTEMPO did not inhibit gastric acid secretion but offered gastroprotection by preventing DCF-induced generation of O2− due to mitochondrial respiratory chain failure and by preventing mitochondrial oxidative stress (MOS)-mediated mitopathology. MitoTEMPO even restored DCF-stimulated reduced fatty acid oxidation, mitochondrial depolarization and bioenergetic crisis in gastric mucosa. MitoTEMPO also prevented the activation of mitochondrial pathway of apoptosis and MOS-mediated proinflammatory signaling through NF-κB by DCF. Furthermore, mitoTEMPO when administered in rats with preformed gastric lesions expedited the healing of gastric injury and the healed stomach exhibited its normal physiology as evident from gastric acid and pepsin secretions under basal or stimulated conditions. Thus, in contrast to the existing antiulcer drugs, mitochondrially targeted O2− scavengers like mitoTEMPO may represent a novel class of gastroprotective molecules that does not affect gastric acid secretion and may be used in combination with DCF, keeping its anti-inflammatory action intact, while reducing its gastrodamaging effects.

Mast cells are associated with the onset and progression of celiac disease

Celiac disease (CD) is an immune-mediated disorder characterized by an accumulation of immune cells in the duodenal mucosa as a consequence of both adaptive and innate immune responses to undigested gliadin peptides. Mast cells (MCs) are innate immune cells that are a major source of costimulatory signals and inflammatory mediators in the intestinal mucosa. Although MCs have previously been associated with CD, functional studies have never been performed. We aimed at evaluating the role of MCs in the pathogenesis of CD. Intestinal biopsy specimens of patients with CD were scored according to the Marsh classification and characterized for leukocyte infiltration and MC distribution. Moreover, MC reactivity to gliadin and its peptides was characterized by using invitro assays. Infiltrating MCs were associated with the severity of mucosal damage, and their numbers were increased in patients with higher Marsh scores. MCs were found to directly respond to nonimmunodominant gliadin fragments by releasing proinflammatory mediators. Immunohistochemical characterization of infiltrating MCs and the effects of gliadin peptides on intestinal MCs indicated an increase in proinflammatory MC function in advanced stages of the disease. This was also associated with increased neutrophil accumulation, the prevalence of M1 macrophages, and the severity of tissue damage. We provide a description of the progressive stages of CD, in which MCs are the hallmark of the inflammatory process. Thus the view of CD should be revised, and the contribution of MCs in the onset and progression of CD should be reconsidered in developing new therapeutic approaches.