Villus Atrophy Research Articles

Porcine Deltacoronavirus Infection Disrupts the Intestinal Mucosal Barrier and Inhibits Intestinal Stem Cell Differentiation to Goblet Cells via the Notch Signaling Pathway.

Goblet cells and their secreted mucus are important elements of the intestinal mucosal barrier, which allows host cells to resist invasion by intestinal pathogens. Porcine deltacoronavirus (PDCoV) is an emerging swine enteric virus that causes severe diarrhea in pigs and causes large economic losses to pork producers worldwide. To date, the molecular mechanisms by which PDCoV regulates the function and differentiation of goblet cells and disrupts the intestinal mucosal barrier remain to be determined. Here, we report that in newborn piglets, PDCoV infection disrupts the intestinal barrier: specifically, there is intestinal villus atrophy, crypt depth increases, and tight junctions are disrupted. There is also a significant reduction in the number of goblet cells and the expression of MUC-2. In vitro, using intestinal monolayer organoids, we found that PDCoV infection activates the Notch signaling pathway, resulting in upregulated expression of HES-1 and downregulated expression of ATOH-1 and thereby inhibiting the differentiation of intestinal stem cells into goblet cells. Our study shows that PDCoV infection activates the Notch signaling pathway to inhibit the differentiation of goblet cells and their mucus secretion, resulting in disruption of the intestinal mucosal barrier. IMPORTANCE The intestinal mucosal barrier, mainly secreted by the intestinal goblet cells, is a crucial first line of defense against pathogenic microorganisms. PDCoV regulates the function and differentiation of goblet cells, thereby disrupting the mucosal barrier; however, the mechanism by which PDCoV disrupts the barrier is not known. Here, we report that in vivo, PDCoV infection decreases villus length, increases crypt depth, and disrupts tight junctions. Moreover, PDCoV activates the Notch signaling pathway, inhibiting goblet cell differentiation and mucus secretion in vivo and in vitro. Thus, our results provide a novel insight into the mechanism underlying intestinal mucosal barrier dysfunction caused by coronavirus infection.

Low-dose nafamostat mesilate ameliorates tissue injury and inhibits 5-hydroxytryptamine synthesis in the rat intestine after methotrexate administration

We aimed to clarify the effect of nafamostat mesilate (nafamostat) on intestinal mucositis as well as the potentiation of intestinal 5-hydroxytryptamine (5-HT) dynamics induced by methotrexate, an anti-cancer drug, in rats. Rats received intraperitoneal methotrexate at 12.5 mg/kg/day for 4 days. In addition, 1, 3, or 10 mg/kg/day of nafamostat was given subcutaneously for 4 days. Ninety-six hours after the first administration of methotrexate, jejunal tissues were collected for analysis. The results showed that 1 mg/kg, but not 3 or 10 mg/kg, of nafamostat significantly ameliorated the methotrexate-induced body weight loss. Moreover, 1 mg/kg of nafamostat significantly improved methotrexate-induced mucositis, including villus atrophy. Nafamostat (1 mg/kg) significantly inhibited the methotrexate-induced mRNA expression of pro-inflammatory cytokines and cyclooxygenase-2, as well as methotrexate-induced 5-HT content and tryptophan hydroxylase (TPH) activity. In addition, it tended to inhibit the number of anti-TPH antibody-positive cells and significantly inhibited the number of anti-substance P antibody-positive cells. These findings suggest that low-dose nafamostat ameliorates tissue injury and 5-HT and substance P synthesis in methotrexate-induced mucositis. Nafamostat may be a novel therapeutic strategy for the prevention and treatment of mucositis as well as 5-HT- and/or substance P-related adverse effects in cancer chemotherapy.

Short-chain fatty acid-releasing nano-prodrugs for attenuating growth and metastasis of melanoma.

Low-molecular-weight (LMW) short-chain fatty acids (SCFAs), such as propionic and butyric acids, have been reported to possess anti-neoplastic effects; however, rapid renal clearance and high dose-based side effects limit their clinical translation. Hence, in this study, we have designed a new self-assembling nano-prodrugs that can effectively supply SCFAs: endogenous enzyme-metabolizable block copolymer poly(ethylene glycol)block-poly(vinyl ester) possessing several units of SCFAs conjugated as side chains via ester linkages. These amphiphilic polymers spontaneously self-assemble into nanostructures under aqueous conditions to form orally administrable nano-prodrugs (butyric acid: NanoBA and propionic acid: NanoPA). Herein, we show the therapeutic efficacy of SCFA nanoparticles (NanoSCFA) in a mouse model of metastasis (melanoma). Ad libitum intake of our NanoSCFA markedly demonstrated a decrease in the metastatic tumor nodules in the lungs compared with the effect observed after LMW SCFA administration with no discernible toxicity to the GI tract. In contrast, LMW SCFAs, even at a lower concentration than that of the NanoSCFA, facilitated villus atrophy. Taken together, our work suggests that the use of NanoSCFA as a therapeutic intervention for metastatic cancer is preferable over typical LMW SCFAs. STATEMENT OF SIGNIFICANCE: Low-molecular-weight (LMW) short-chain fatty acids (SCFAs) have shown versatile therapeutic effects on various diseases, including anti-tumorigenesis effects. However, their clinical translation is limited due to their poor pharmacokinetic profile and adverse effects. To overcome these limitations, we have developed new amphiphilic block copolymer-based SCFA-prodrugs, which self-assemble into nanoparticles in aqueous media (NanoSCFA). SCFAs are covalently conjugated to the hydrophobic polymer segment via ester linkage, which can be enzymatically metabolized after oral administration. In the present study, we confirmed that ad libitum intake of NanoSCFAs retarded the growth and metastatic potential of B16-F10 tumors compared to the LMW SCFAs with negligible discernible toxicity, reflecting NanoSCFA as a preferable therapeutic intervention to LMW SCFA counterparts.

Quercetin attenuates deoxynivalenol-induced intestinal barrier dysfunction by activation of Nrf2 signaling pathway in IPEC-J2 cells and weaned piglets

The presence of deoxynivalenol (DON), one of the most frequently occurring mycotoxin, in food and feed has been considered a risk factor to both human and animal health. Molecular mechanisms that regulate DON effects in tissues are still poorly understood. However, recent evidence suggests that nuclear factor erythroid 2-like 2 (Nrf2) may be a major target during mycotoxin-induced intestinal barrier dysfunction. Although quercetin, a plant-derived flavonoid, is known to induce the activation of Nrf2 signaling pathway, its potential to mitigate effects of DON and the implication of Nrf2 in its physiological effects is poorly understood. Therefore, this study was conducted to investigate the protective effects of quercetin in alleviating the DON-induced barrier loss and intestinal injuries in IPEC-J2 cells and weaned piglets and determine the potential role of Nrf2. Quercetin treatment dose-dependently increased mRNA expression of Nrf2 target gene, NQO-1, and concomitantly increased the expression of claudin-4 at both mRNA and protein levels. Quercetin supplementation also reversed the reduction of claudin-4 caused by DON exposure in vivo and in vitro. The decreased membrane presence of claudin-4 and ZO-1 induced by DON was also blocked by quercetin. Furthermore, quercetin attenuated the endocytosis and degradation of claudin-4 caused by DON exposure. The effects of quercetin also included the restoration of transepithelial electrical resistance (TEER) and reduction of FITC-dextran permeability that have been perturbed by DON. However, the protective effects of quercetin against DON exposure were abolished by a specific Nrf2 inhibitor (brusatol), confirming the importance of Nrf2 in the regulation of TJP expression and barrier function by quercetin. In vivo study in weaned pigs showed that DON exposure impaired villus-crypt morphology as indicated by diffuse apical villus necrosis, villus atrophy and fusion. Notably, intestinal injuries caused by DON administration were partly mitigated by quercetin supplementation. Collectively, this study shows that quercetin could be used to prevent the DON-induced gut barrier dysfunction in humans and animals and the protective effects of quercetin against DON-induced intestinal barrier disruption is partly through Nrf2-dependent signaling pathway.

Role of Bifidobacterium infantis in the Treatment of Duodenal and Colon Inflammation in Induced Ulcerative Colitis


 
 
 
 This study aimed to investigate the potential protective role of Bifidobacterium (B.) infantis in alleviating ‎‎induced duodenal and colon inflammation associated with ulcerative colitis (UC). Female albino Wister rats ‎‎(n=24) ‎were randomized into four experimental groups: Control Negative, acetic acid-induced colitis (AA colitis), ‎B. infantis-treated (Bifido), orally gavaged with 1 mL of reference strain ‎B. infantis at 108 CFU/mL for four weeks prior to the induction of colitis,‎ and Bifido+AA ‎colitis. Colitis was induced via ‎intrarectal administration of 4% AA solution‎. Seven days post-colitis induction, blood samples were ‎obtained to assess protein levels, and histopathological evaluations were conducted on duodenal and ‎colon tissues. ‎Additionally, immunohistochemical assessments for B-cell lymphoma 2‎ (Bcl-2) in colon and ‎myeloperoxidase (MPO) in duodenum ‎sections were performed‎. Results revealed that B. infantis treatment significantly elevated serum albumin and total ‎protein levels in the Bifido and Bifido+AA colitis groups, approximating those in the Control Negative group. Histopathological and morphological changes of duodenum in AA colitis revealed ulceration of the mucosal epithelium, submucosal inflammatory cellular infiltration, tissue depression resulting in villus atrophy, and crypt hyperplasia. Additionally, ‎colonic crypt gland atrophy and goblet cells depletion were observed. Most of these changes were ameliorated in the Bifido and Bifido+AA colitis groups. Immunohistochemical analysis displayed marked immunopositivity ‎ of Bcl-2 in colon and MPO in duodenum sections of the Bifido and Bifido+AA colitis groups, indicating the antiapoptotic and anti-inflammatory roles of B. infantis. This study demonstrates that B. infantis exerts a protective effect against AA-induced UC by normalizing serum protein levels, ameliorating histopathological alterations, and ‎modulating apoptotic and inflammatory markers. These findings underscore B. infantis as a ‎promising therapeutic agent for UC and warrant further research to elucidate the underlying ‎molecular mechanisms.
 
 
 

Impaired FXR-CPT1a signaling contributes to parenteral nutrition-induced villus atrophy in short-bowel syndrome.

Parenteral nutrition (PN)-induced villus atrophy is a major cause of intestinal failure (IF) for children suffering from short bowel syndrome (SBS), but the precise mechanism remains unclear. Herein, we report a pivotal role of farnesoid X receptor (FXR) signaling and fatty acid oxidation (FAO) in PN-induced villus atrophy. A total of 14 pediatric SBS patients receiving PN were enrolled in this study. Those patients with IF showed longer PN duration and significant intestinal villus atrophy, characterized by remarkably increased enterocyte apoptosis concomitant with impaired FXR signaling and decreased FAO genes including carnitine palmitoyltransferase 1a (CPT1a). Likewise, similar changes were found in an in vivo model of neonatal Bama piglets receiving 14-day PN, including villus atrophy and particularly disturbed FAO process responding to impaired FXR signaling. Finally, in order to consolidate the role of the FXR-CPT1a axis in modulating enterocyte apoptosis, patient-derived organoids (PDOs) were used as a mini-gut model in vitro. Consequently, pharmacological inhibition of FXR by tauro-β-muricholic acid (T-βMCA) evidently suppressed CPT1a expression leading to reduced mitochondrial FAO function and inducible apoptosis. In conclusion, impaired FXR/CPT1a axis and disturbed FAO may play a pivotal role in PN-induced villus atrophy, contributing to intestinal failure in SBS patients.

Evaluation and validation of chemotherapy-specific diarrhoea and histopathology in rats.

Chemotherapy-induced mucositis is characterized by diarrhoea and villous atrophy. However, it is not well-understood why diarrhoea arises, why it only occurs with some chemotherapeutics and how it is related to villus atrophy. The objectives in this study were to determine (i) the relationship between chemotherapy-induced diarrhoea and villus atrophy and to (ii) establish and validate a rat diarrhoea model with clinically relevant endpoints. Male Wistar Han IGS rats were treated with saline, doxorubicin, idarubicin, methotrexate, 5-fluorouracil, irinotecan or 5-fluorouracil+irinotecan. After 72 h, jejunal tissue was taken for morphological, apoptotic and proliferative analyses, and faecal water content and change in body weight were determined. All treatments except methotrexate caused a similar reduction (≈42%) in villus height, but none of them altered mucosal crypt cell proliferation or apoptosis. Doxorubicin, idarubicin, irinotecan and 5-fluorouracil+irinotecan caused body weight reduction, but only irinotecan and idarubicin caused diarrhoea. No direct correlation between diarrhoea and villus height or body weight loss was observed. Therefore, studies of the mechanisms for chemotherapy-induced diarrhoea should focus on functional factors. Finally, the irinotecan and idarubicin diarrhoea models established in this study will be useful in developing supportive treatments of this common and serious adverse effect in patients undergoing chemotherapy.

A Comparative Study on Pathological Changes in the Small Intestine of Sheep and Goat Experimentally Infected with Trichostrongylus colubriformis.

BackgroundTrichostrongylus colubriformis, also called hairworm, is a genus of parasitic roundworm affecting gastro-intestinal tracts of a ruminant. Gross and microscopic lesion characterizations and comparing its effect in the small intestine of sheep and goats experimentally infected with T. colubriformis were undertaken in the study.MethodsDuring the study period, 13 sheep and 14 goats were included in the experiment. The larvae of T. culibriformis were obtained from abattoirs and larvae were recovered by Bearmann techniques. The infective larvae of T. culibriformis (L3) as a single dose of 10,000 per-animal was administered orally to infected groups of sheep and goats. Blood was collected for hematological and serum biochemical analysis. Tissues for gross and histopathologic lesions characterization were collected from killed infected animals at 56 days.ResultsFrom the infected group, the total recovered mean worm burden was recorded as higher in goats (P<0.05) than sheep, with an establishment rate of 50.16% and 34.46%, respectively. The total mean PCV, Hb, and albumin values recorded in the infected groups of sheep and goats were significantly (P<0.05) lower than non-infected control of both animal groups. In goats, the total serum protein was significantly (P<0.05) lower in the infected group than the non-infected control group. Gross lesions found were enteritis with petechial hemorrhages, edema, hyperemia, and mucosal slough, which were marked in the duodenum (62.69%) and jejunum (33.33%) in sheep and 47.05% duodenum and 45.09% jejunum in goats. The microscopic lesions developed by T. colubriformis were subtotal villus atrophy, hemorrhage, straightened and elongated dilated crypts, loss of epithelium, mucosal erosion, and infiltration of inflammatory cells.ConclusionThe present study showed that T. colubriformis infection caused physiological and pathological changes of the small-intestine in sheep and goats, with more severe infection in goats than sheep, although they were under the same management condition.

Shenhuang Plaster Application Improves Gastrointestinal Motility in Mice with Postoperative Ileus through Intestinal Microbiota.

Postoperative ileus (POI) is a common surgical complication, and its incidence remains high. Shenhuang Plaster (SHP) is a famous traditional Chinese medicine with a definite curative effect on postoperative intestinal dysfunction; however, the mechanisms involved in these effects are unclear. Accordingly, in this study, we constructed a POI mouse model and used the intestinal flora as the target to explore the regulatory effect of SHP on gastrointestinal motility. The results illustrated that SHP applied at the Shenque acupoint promoted the recovery of gastrointestinal motility, relieved intestinal villus atrophy and basal damage caused by POI, protected the integrity of intestinal tissue morphology, and alleviated the inflammatory response in the intestinal tissue of POI model mice. In addition, we clarified the role of the intestinal flora in the occurrence and development of POI, further evaluated the changes in the intestinal flora in each group of mice, and analysed the regulatory effect of SHP on the intestinal flora in mice with POI. The results suggested that SHP might improve gastrointestinal motility disorder in POI mice by effectively regulating intestinal flora.

Tocopherol Enhances the Antioxidant Defense System and Histomorphometric Parameters in The Gastrointestinal Tract of Rats Treated with Sodium Arsenite.

Arsenic compromises the gastrointestinal integrity and function via the body's anti-oxidative system breakdown. Hence, this study aimed to investigate the effects of tocopherol on redox imbalance and histoarchitectural alterations in rats' gastrointestinal tract exposed to sodium arsenite. Sodium arsenite and graded doses of tocopherol were administered orally into experimental rats assigned to different groups for four weeks concurrently. Redox status assay was done in homogenized samples by spectrophotometry. Parietal cell mass and mucous cell density (stomach), villus height and crypt depth (ileum), goblet cells count, and crypt depth (colon) were evaluated by histomorphometry. Inflammatory cells infiltration was also assessed using a semi-quantitative procedure.Sodium arsenite caused a significant increase in Malondialdehyde and Myeloperoxidase but, decreased Superoxide dismutase, Catalase, Nitric oxide, Glutathione peroxidase, Glutathione, and Glutathione-S-Transferase. Tocopherol treatment reversed the changes (p<0.05) though not largely dose-dependent. Furthermore, tocopherol annulled sodium arsenite-induced increase in parietal cell mass and decrease in mucous cell density in the stomach, decrease in villus height and villus height/crypt depth ratio in the ileum, and decrease in goblets cells and increase in crypt depth in the colon. Moreover, activated inflammatory cell infiltration by sodium arsenite was mitigated by tocopherol. Sodium arsenite provokes not only marked inflammatory cellular infiltration but a focal loss of glands, hyperplasia of crypts, atrophic villi, and hypertrophy of Peyer's patches in the intestines, which are all lessened with tocopherol treatment. These findings underscore the anti-oxidative properties of tocopherol as a potent dietary factor against sodium arsenite toxicity in the gastrointestinal tract. Keywords: Tocopherol, arsenic, stomach, ileum, colon.

Factor-Based Private Debt Investing at the Industry and Country Level

<h3>OBJECTIVES:</h3> To investigate the effects of enteral and parenteral glutamine (Gln) usage on rats in sepsis. <h3>METHODS:</h3> This study was conducted in Istanbul University Experimental Medical Research Institution (DETAE) laboratory, Istanbul University, Istanbul, Turkey between June and September 2009. The levels of blood Gln, tumor necrosis-alpha (TNF-alpha) and interleukin (IL)-10 was measured. Samples of tissue were obtained from the mesenteric lymph nodes, liver, and lower lobe of the right lung to evaluate the reproduction of bacteria, and samples of tissue were also obtained from the small intestine to evaluate blood and villus atrophy. <h3>RESULTS:</h3> Bacteremia of Group EP (combined group) were found lower than Group C (control) (p=0.007). Villous atrophy rates of all groups were lower than Group C: between Group E (enteral) and C (p=0.003); between Group P (parenteral alanine) and C (p=0.019); and between Group EP and C (p=0.001). The values of serum TNF-alpha and IL-10 of Group EP and P were lower than the other groups at the 24th and 96th hours (p=0.000). <h3>CONCLUSIONS:</h3> In this study we found that the most efficient Gln administration technique in sepsis was enteral administration together with parenteral administration due to trophic effect on the intestinal mucosa, decrease of reproduction in tissue and blood cultures, immunomodulator effect, and approximately the same cost as parenteral application.

Relationship between serum cobalamin concentration and endoscopic ileal appearance and histology in dogs with chronic inflammatory enteropathy.

BackgroundIt has not been determined whether ileal appearance differs among dogs with chronic inflammatory enteropathy (CIE) and different serum concentrations of cobalamin.ObjectiveTo compare endoscopic and histologic ileal findings in dogs with CIE and different serum cobalamin concentrations and then evaluate the correlation of ileal changes to cobalamin serum concentration using updated scoring systems to assess the ileum.AnimalsSixty‐eight dogs with CIE.MethodsRetrospective study. Frequency of ileal features and ileal histologic and endoscopic scores (IHS and IES) were obtained and compared among CIE dogs with severe hypocobalaminemia (SHC; <200 ng/L), hypocobalaminemia (HC; 200‐350 ng/L), or normocobalaminemia (NC; >350 ng/L). The correlation of IHS and IES with cobalamin was evaluated.ResultsFriability, villus atrophy, crypt dilatation, epithelial injury, and intraepithelial lymphocytes were more frequent in SHC than in NC dogs (all P ≤ .01). Median SHC‐IES (2; range, 0‐4) was higher than NC‐IES (1; range, 0‐5; P = .004). Median SHC‐IHS (6; range, 3‐9) was higher than HC‐IHS (4; range, 1‐7; P < .001) and NC‐IHS (3; range, 1‐8; P < .001). Cobalamin concentration correlated negatively with IES (ρ = −.34, P = .005) and IHS (ρ = −.58, P < .001).Conclusions and Clinical ImportanceIleal features and involvement degree markedly differed when cobalamin was <200 or >350 ng/L in CIE dogs. With updated scales to assess the mucosa, greater ileal damage was associated with lower serum cobalamin concentration.

Lactobacillus delbrueckii CIDCA 133 Ameliorates Chemotherapy-Induced Mucositis by Modulating Epithelial Barrier and TLR2/4/Myd88/NF-κB Signaling Pathway

Intestinal mucositis promoted by the use of anticancer drugs is characterized by ulcerative inflammation of the intestinal mucosa, a debilitating side effect in cancer patients undergoing treatment. Probiotics are a potential therapeutic option to alleviate intestinal mucositis due to their effects on epithelial barrier integrity and anti-inflammatory modulation. This study investigated the health-promoting impact of Lactobacillus delbrueckii CIDCA 133 in modulating inflammatory and epithelial barrier markers to protect the intestinal mucosa from 5-fluorouracil-induced epithelial damage. L. delbrueckii CIDCA 133 consumption ameliorated small intestine shortening, inflammatory cell infiltration, intestinal permeability, villus atrophy, and goblet cell count, improving the intestinal mucosa architecture and its function in treated mice. Upregulation of Muc2, Cldn1, Hp, F11r, and Il10, and downregulation of markers involved in NF-κB signaling pathway activation (Tlr2, Tlr4, Nfkb1, Il6, and Il1b) were observed at the mRNA level. This work suggests a beneficial role of L. delbrueckii strain CIDCA 133 on intestinal damage induced by 5-FU chemotherapy through modulation of inflammatory pathways and improvement of epithelial barrier function.

The effects of creep feed composition and form and nursery diet complexity on small intestinal morphology and jejunal mucosa-specific enzyme activities after weaning in pigs.

Fifty-six litters from first-parity sows standardized to 12 piglets were used to determine the effects of creep feed composition and form and the provision of low- or high-complexity nursery diets on the evolution of small intestinal histomorphology and jejunal mucosa-specific enzyme activities postweaning. At 5 d of age, litters (initial bodyweight [BW] 2.31 ± 0.61 kg) were assigned to one of four creep feeding regimens (n = 14): 1) commercial creep feed (COM), 2) liquid milk replacer (LMR), 3) pelleted milk replacer (PMR), or 4) no creep feed (NO). At weaning (21 d of age), six pigs per litter were provided a HIGH- (contained highly digestible animal proteins) or LOW- (contained corn and soybean meal as main protein sources) complexity nursery diet (n = 7). At 21, 28, and 59 d of age, two pigs per pen (one castrated male and one female) were euthanized, and ileal and jejunal segments for histomorphological measurements and jejunal mucosal scrapings were collected to determine specific mucosa enzyme activities. At weaning, pigs provided COM had a greater ileal absorptive capacity (M) than LMR or NO, which were not different (14.1 vs. 10.4 and 10.5 ± 0.9 μm2; P < 0.05); PMR was intermediate. On days 28 and 59, M was not different among pigs regardless of creep feed treatments. Pigs fed LOW had reduced jejunal villus height (VH; P < 0.001) and M (P < 0.001) on day 28 vs. day 21. The VH and M were not different for pigs fed HIGH or LOW by the end of the nursery period. For all dietary treatments except COM-HIGH and COM-LOW, jejunal mucosal maltase-specific activity was not different between days 21 and 28 of age but greater on day 59 (P < 0.05). For pigs that received COM-HIGH, maltase-specific activity was not different between days 21 and 28 but greater on day 59 than day 28 (P < 0.05). For pigs that received COM-LOW, maltase-specific activity was not different between days 21, 28, and 59. Regardless of creep or nursery treatment, sucrase-specific activity was the greatest on day 59, followed by days 21 and 28 (P < 0.001), and lactase-specific activity was greater on day 21 than on days 28 and 59 (P < 0.001), which were not different. Therefore, pigs that provided LOW diet had greater villus atrophy and reduced M during the first week after weaning vs. pigs that provided HIGH, regardless of creep feeding regimen, but were able to recover by the end of the nursery period.

Protective role of casuarinin from Melastoma malabathricum against a mouse model of 5-fluorouracil–induced intestinal mucositis: Impact on inflammation and gut microbiota dysbiosis

Background5-FU-induced intestinal mucositis (FUIIM) is a common gastrointestinal side effect of chemotherapy, leading to gastric pain in clinical cancer patients. In a previous study, we demonstrated that neutrophil elastase (NE) inhibitors could alleviate FUIIM and manipulate the homeostasis of the gut microbiota. The root of Melastoma malabathricum, also called Ye-Mu-Dan, has been used as a traditional Chinese medicine for gastrointestinal disease. Water extract of the roots of M. malabathricum exhibits an inhibitory effect on NE, with an IC50 value of 9.13 μg/ml. PurposeIn this study, we aimed to isolate an anti-NE compound from the root of M. malabathricum and to determine the protective effect of the bioactive component on a mouse model of FUIIM with respect to tissue damage, inflammation, intestinal barrier dysfunction, and gut microbiota dysbiosis. MethodsA water extract of the roots of M. malabathricum was prepared and its major bioactive compound, was identified using bioactivity-guided fractionation. The effects of samples on the inhibition of NE activity were evaluated using enzymatic assays. To evaluate the effects of the bioactive compound in an FUIIM animal model, male C57BL/6 mice treated with or without casuarinin (50 and 100 mg/kg/day, p.o.), and then received of 5-fluorouracil (50 mg/kg/day) intraperitoneally for 5 days to induce FUIIM. Histopathological staining was used to monitor the tissue damage, proliferation of intestinal crypts, and expression of tight junction proteins. The inflammation score was estimated by determining the levels of oxidative stress, neutrophil-related proteases, and proinflammatory cytokines in tissue and serum. The ecology of the gut microbiota was evaluated using 16S rRNA gene sequencing. ResultsCasuarinin had the most potent and selective effect against NE, with an IC50 value of 2.79 ± 0.07 μM. Casuarinin (100 mg/kg/day, p.o.) significantly improved 5-FU-induced body weight loss together with food intake reduction, and it also significantly reversed villus atrophy, restored the proliferative activity of the intestinal crypts, and suppressed inflammation and intestinal barrier dysfunction in the mouse model of FUIIM. Casuarinin also reversed 5-FU-induced gut microbiota dysbiosis, particularly the abundance of Actinobacteria, Candidatus Arthromitus, and Lactobacillus murinus, and the Firmicutes-to-Bacteroidetes ratio. ConclusionThis study firstly showed that casuarinin isolated from the root part of M. malabathricum could be used as a NE inhibitor, whereas it could improve FUIIM by modulating inflammation, intestinal barrier dysfunction, and gut microbiota dysbiosis. In summary, exploring anti-NE natural product may provide a way to find candidate for improvement of FUIIM.

Single-Cell RNA Sequencing of Peripheral Blood Mononuclear Cells From Pediatric Coeliac Disease Patients Suggests Potential Pre-Seroconversion Markers.

Celiac Disease (CeD) is a complex immune disorder involving villous atrophy in the small intestine that is triggered by gluten intake. Current CeD diagnosis is based on late-stage pathophysiological parameters such as detection of specific antibodies in blood and histochemical detection of villus atrophy and lymphocyte infiltration in intestinal biopsies. To date, no early onset biomarkers are available that would help prevent widespread villous atrophy and severe symptoms and co-morbidities. To search for novel CeD biomarkers, we used single-cell RNA sequencing (scRNAseq) to investigate PBMC samples from 11 children before and after seroconversion for CeD and 10 control individuals matched for age, sex and HLA-genotype. We generated scRNAseq profiles of 9559 cells and identified the expected major cellular lineages. Cell proportions remained stable across the different timepoints and health conditions, but we observed differences in gene expression profiles in specific cell types when comparing patient samples before and after disease development and comparing patients with controls. Based on the time when transcripts were differentially expressed, we could classify the deregulated genes as biomarkers for active CeD or as potential pre-diagnostic markers. Pathway analysis showed that active CeD biomarkers display a transcriptional profile associated with antigen activation in CD4+ T cells, whereas NK cells express a subset of biomarker genes even before CeD diagnosis. Intersection of biomarker genes with CeD-associated genetic risk loci pinpointed genetic factors that might play a role in CeD onset. Investigation of potential cellular interaction pathways of PBMC cell subpopulations highlighted the importance of TNF pathways in CeD. Altogether, our results pinpoint genes and pathways that are altered prior to and during CeD onset, thereby identifying novel potential biomarkers for CeD diagnosis in blood.

A233 CHARACTERIZATION OF THE DUODENAL MICROBIOME IN PEDIATRIC CELIAC AND INFLAMMATORY BOWEL DISEASE PATIENTS

BackgroundBoth Celiac disease (CE) and inflammatory bowel disease (IBD) are lifelong gastrointestinal tract (GIT) disorders. CE is an autoimmune disorder triggered by gluten consumption and can result in small intestine villus atrophy, crypt hyperplasia and epithelial permeability, while IBD is characterized by chronic, reoccurring inflammation and ulcers in the GIT. There is increasing evidence linking GIT microbes to both CE and IBD pathogenesis. Studies have also shown that CE patients are at increased risk of developing IBD, suggesting a possible link between these diseases. The duodenum can be affected in both CE and IBD, but it is understudied as compared to other GIT regions.AimsWe thus aimed to characterize the duodenal microbiome of pediatric CE, IBD and control patients (n=76, 48 and 57 respectively) and hypothesized that the composition of microbes would vary between each diagnostic group.MethodsWe used mucosal luminal interface aspirates collected during upper endoscopy at the Children’s Hospital of Eastern Ontario. Metagenomic DNA was extracted from these samples and bacterial taxa was characterized by sequencing the V6 hypervariable region of the 16S rRNA gene.ResultsControl, CE, and IBD duodenal microbiotas showed no apparent differences in either α-diversity or β-diversity. However, we identified several significant differences between the relative abundances of specific taxa in these three patient groups. In particular, Atopobium parvulum was found to be enriched in Crohn’s disease samples when compared to non-IBD controls. There was also a trend for higher Bacteroides, Lactobacillus, Parabacteroides and Staphylococcus in CE patient MLI aspirates. These results are similar to what has been previously reported in CE patients. Finally, trends found in the duodenal MLI aspirates are more consistent with results previously found in the salivary microbiome in CD patients as opposed to studies of the large intestine.ConclusionsThis work provides unique insight into the microbial composition at the duodenum; a GIT region that has not been fully characterized in CE or IBD.Funding AgenciesCIHRGenome Canada

Rotavirus Interactions With Host Intestinal Epithelial Cells.

Rotavirus (RV) is the foremost enteric pathogen associated with severe diarrheal illness in young children (<5years) and animals worldwide. RV primarily infects mature enterocytes in the intestinal epithelium causing villus atrophy, enhanced epithelial cell turnover and apoptosis. Intestinal epithelial cells (IECs) being the first physical barrier against RV infection employs a range of innate immune strategies to counteract RVs invasion, including mucus production, toll-like receptor signaling and cytokine/chemokine production. Conversely, RVs have evolved numerous mechanisms to escape/subvert host immunity, seizing translation machinery of the host for effective replication and transmission. RV cell entry process involve penetration through the outer mucus layer, interaction with cell surface molecules and intestinal microbiota before reaching the IECs. For successful cell attachment and entry, RVs use sialic acid, histo-blood group antigens, heat shock cognate protein 70 and cell-surface integrins as attachment factors and/or (co)-receptors. In this review, a comprehensive summary of the existing knowledge of mechanisms underlying RV-IECs interactions, including the role of gut microbiota, during RV infection is presented. Understanding these mechanisms is imperative for developing efficacious strategies to control RV infections, including development of antiviral therapies and vaccines that target specific immune system antagonists within IECs.

Morphological and histopathological studies of Thelandros chalcidae (Oxyuroidea: Pharyngodonidae) infecting Chalcides ocellatus from Egypt

BackgroundThelandros (Pharyngodonidae) is a gastrointestinal nematode parasite with a life cycle including lizards as main hosts. Thelandros chalcidae collected from the large intestine of the Egyptian ocellated skink, Chalcides ocellatus were described and illustrated by light and scanning electron microscopes. Seven out of fifteen (46.66%) of the examined lizards were found to be naturally infected. Also, host intestinal tissues were evaluated from hematoxylin/eosin-stained sections to describe any histopathological changes.ResultsMicroscopic examinations revealed that the recovered pharyngodonid species characterized by mouth with triangular opening and surrounded by six simple lips, the cuticle had regular transverse annulations extending from the posterior margin of the lips to the end of the body. Male was cylindrical with distinct truncated posterior end and measured 1.59–1.86 (1.64 ± 0.10) long and 0.29–0.37 (0.32 ± 0.01) in maximum width at the level of mid-body. Female measured 1.72–2.43 (1.85 ± 0.2) long and 0.36–0.49 (0.42 ± 0.01) maximum width at the mid-body level, terminated posteriorly in a short, stout spike. Histological studies observed structural alterations represented by leukocytic infiltration, villi atrophy, and muscularis degeneration. These changes were indicative of inflammatory and degenerative reaction due to Thelandros chalcidae infection.ConclusionThe present morphological study revealed that the recovered pharyngodonid species was Thelandros chalcidae causing pathological alterations in Chalcides ocellatus intestinal tissues.

CASE REPORT: COEXISTENCE OF CELIAC DISEASE AND MULTIPLE MYELOMA

Celiac disease is a systemic disease in which the natural and adaptive immune system is affected by the effect of gluten exposure and environmental factors in individuals with genetic predisposition. Multiple myeloma; is characterized by an increase in clonal plasma cells. It is the most common hematological malignancy after lymphomas.We aimed to present a case siagnosed with celiac disease and multipl myeloma A 56-year-old female patient with a diagnosis of asthma and celiac disease for 1 year was referred to the Hematology department because her refractory anemia. Serum IgA level of the patient was 4490 mg/dl without renal failure and hypercalcemia.bone marrow biopsy compatible with myeloma. The patient received 6 cycles of bortezomib, cyclophosphamide, and dexamethasone and 3 cycles lenalidomid dexametazon chemotherapy.After chemotherapy, Autologous stem cell transplantation was performed. Celiac disease is an autoimmune disease, characterized by inflammation and villus atrophy in the small intestine mucosa as a result of sensitivity to gluten, resulting in malabsorption. The incidence of lymphoma and gastrointestinal system malignancy is increased in individuals with celiac disease. Multiple myeloma may also be accompanied by autoimmune diseases such as ankylosing spondylitis, scleroderma, and sjögren's syndrome. Coexistence of multiple myeloma and celiac disease is rare.