Intestinal Architecture Research Articles

Protective effects of the fructooligosaccharide on the growth performance, biochemical indexes, and intestinal morphology of blunt snout bream (Megalobrama amblycephala) infected by Aeromonas hydrophila.

The purpose of the study was to investigate the effects of dietary fructooligosaccharide (FOS) on growth performance, biochemical indexes, intestinal morphology, and growth-related gene expression of blunt snout bream (Megalobrama amblycephala) infected by Aeromonas hydrophila (AH). Two hundred twenty-five healthy blunt snout bream with an initial body weight of 38.41 ± 0.88g were randomly divided into five groups with three replicates: control (basal diet), model (AH + basal diet), SFOS (AH + 2g/kg FOS), MFOS (AH + 4g/kg FOS), LFOS (AH + 6g/kg FOS). After 9weeks of feeding, the results showed that the FOS-added diet abrogated AH-induced retardation, hemorrhage, and inflammatory infiltration. FOS supplementation enhanced the growth performance degradation caused by AH, and the highest growth performance was observed at MFOS. Meanwhile, the addition of FOS to feed improved the blood immunity reduced by AH. In expansion, the mucosal epithelium of intestinal villi exfoliated, exposing the lamina propria, and a few villi were genuinely harmed in the model group. Fish fed with MFOS ameliorated the damaged intestine, evidenced by well-preserved intestine architecture. Furthermore, the model group downregulated the expression of growth-related genes (growth hormone receptor (GHR), insulin-like growth factor 1 (IGF-1)). Fish fed with 2g/kg or 4g/kg FOS upregulated the genes specified above expressions in the liver compared with the model group. In conclusion, the results mentioned above suggested that the dietary FOS could relieve the pressure to elevate the immune damage and intestine injury induced by AH and enhance the hepatic expression of IGF-1 and GHR.

Glycoprotein (GP)96 Is Essential for Maintaining Intestinal Epithelial Architecture by Supporting Its Self-Renewal Capacity

Glycoprotein (GP)96 is an endoplasmic reticulum-resident master chaperone for cell surface receptors including the Wnt co-receptors low-densitylipoprotein-receptor-related protein 5/6. Intestinal epithelial cell (IEC)-specific deletion of Gp96 is embryonically lethal. However, the role of GP96 in adult intestinal tissue and especially within the intestinal stem cell (ISC) niche is unknown. Here, we investigated how GP96 loss interferes with intestinal homeostasis by compromising viability, proliferation, and differentiation of IECs. Tamoxifen was used to induce Cre-mediated deletion of Gp96 in GP96-VillincreERT2 (Cre recombinase-Estrogen-Receptor Transgene 2) mice and intestinal organoids. With H&E and immunofluorescence staining we assessed alterations in intestinal morphology and the presence and localization of IEC types. Real-time polymerase chain reaction and Western blot analysis were performed to explore the molecular mechanisms underlying the severe phenotype of Gp96 KO mice and organoids. IEC-specific deletion of Gp96 in adult mice resulted in a rapid degeneration of the stem cell niche, followed by complete eradication of the epithelial layer and death within a few days. These effects were owing to severe defects in ISC renewal and premature ISC differentiation, which resulted from defective Wnt and Notch signaling. Furthermore, depletion of GP96 led to massive induction of endoplasmic reticulum stress. Although effects on ISC renewal and adequate differentiation were partly reversed upon activation of Wnt/Notch signaling, viability could not be restored, indicating that reduced viability was mediated by other mechanisms. Our work shows that GP96 plays a fundamental role in regulating ISC fate and epithelial regeneration and therefore is indispensable for maintaining intestinal epithelial homeostasis.

Adaptation of the gut holobiont to malnutrition during mouse pregnancy depends on the type of nutritional adversity

Malnutrition can influence maternal physiology and programme offspring development. Yet, in pregnancy, little is known about how dietary challenges that influence maternal phenotype affect gut structure and function. Emerging evidence suggests that interactions between the environment, multidrug resistance (MDR) transporters and microbes may influence maternal adaptation to pregnancy and regulate fetoplacental development. We hypothesized that the gut holobiont (host and microbes) during pregnancy adapts differently to suboptimal maternal diets, evidenced by changes in the gut microenvironment, morphology, and expression of key protective MDR transporters during pregnancy. Mice were fed a control diet (CON) during pregnancy, or undernourished (UN) by 30% of control intake from gestational day (GD) 5.5–18.5, or fed 60% high fat diet (HF) for 8 weeks before and during pregnancy. At GD18.5, maternal small intestinal (SI) architecture (H&E), proliferation (Ki67), P-glycoprotein (P-gp - encoded by Abcb1a/b) and breast cancer resistance protein (BCRP/Abcg2) MDR transporter expression and levels of pro-inflammatory biomarkers were assessed. Circulating inflammatory biomarkers and maternal caecal microbiome composition (G3 PhyloChipTM) were measured. MDR transporter expression was also assessed in fetal gut. HF diet increased maternal SI crypt depth and proinflammatory load, and decreased SI expression of Abcb1a mRNA, whilst UN increased SI villi proliferation and Abcb1a, but decreased Abcg2, mRNA expression. There were significant associations between Abcb1a and Abcg2 mRNA levels with relative abundance of specific microbial taxa. Using a systems physiology approach we report that common nutritional adversities provoke adaptations in the pregnancy holobiont in mice, and reveal new mechanisms that could influence reproductive outcomes and fetal development.

PSXIII-15 Effects of Bovine Colostrum Administration on Piglet Growth and Intestinal Health During the Lactation and Post-Weaning Periods

Abstract This study aimed to evaluate the potential of administering bovine colostrum to suckling and weanling piglets to modulate the growth, body composition, intestinal architecture, immunity, as well as remodeling and permeability. Twenty-four litters adjusted to 12 piglets each were selected at birth; one-half of them received liquid bovine colostrum (LC+) from d 5 to d 10, whereas the remaining litters did not receive any supplementation (LC-). Starting from d 11, all the litters received a creep feed either supplemented or not with 5% dehydrated bovine colostrum (BC+ and BC-, respectively). Animals were classified as “eaters” or “non eaters” according to their activity towards the feeders recorded by direct observation and presence of dietary marker (ferric oxide) in feces. At weaning (d 21), 16 LC+/BC+ and 16 LC-/BC- piglets (with an equal number of "eaters" and "non eaters" in each group) were selected and fed for 10 days with the same diets administered during lactation. Growth performance was recorded at d 0, 21 and 31. At d 31 bone mineral content, total body fat and lean contents were measured by dual X-ray absorptiometry; furthermore, animals were euthanized at d 31 and intestinal samples were collected to evaluate the intestinal morphology and concentration of TNFα, IL10, CXCL10, occludin, caspase 3 and proliferating cell nuclear antigen. Our results showed that LC+ and BC+ treatments increased creep feed consumption during lactation (P=0.01 and P=0.08, respectively) and affected litter weight at weaning (LC×BC interaction, P=0.04). Ten days after weaning, BC+ supplementation increased the intestinal villous to crypt ratio and reduced the occludin concentration (P<0.05). Piglets classified as creep feeding "eaters" showed greater intestinal occludin concentrations compared with "non eaters" animals (P=0.005). This study highlights the potential of bovine colostrum to increase piglet growth performance during lactation and affect the intestinal health in the peri-weaning period.

Transit-amplifying cells control R-spondins in the mouse crypt to modulate intestinal stem cell proliferation.

Intestinal epithelium regenerates rapidly through proliferation of intestinal stem cells (ISCs), orchestrated by potent mitogens secreted within the crypt niche. However, mechanisms regulating these mitogenic factors remain largely unknown. Here, we demonstrate that transit-amplifying (TA) cells, marked by unconventional prefoldin RPB5 interactor (URI), control R-spondin production to guide ISC proliferation. Genetic intestinal URI ablation in mice injures TA cells, reducing their survival capacity, leading to an inflamed tissue and subsequently decreasing R-spondin levels, thereby causing ISC quiescence and disruption of intestinal structure. R-spondin supplementation or restoration of R-spondin levels via cell death inhibition by c-MYC elimination or the suppression of inflammation reinstates ISC proliferation in URI-depleted mice. However, selective c-MYC and p53 suppression are required to fully restore TA cell survival and differentiation capacity and preserve complete intestinal architecture. Our data reveal an unexpected role of TA cells, which represent a signaling platform instrumental for controlling inflammatory cues and R-spondin production, essential for maintaining ISC proliferation and tissue regeneration.

Evaluation of methyl orange adsorption potential of green synthesized chitosan-silver nanocomposite (CS–AgNC) and its notable biocompatibility on freshwater Tilapia (Oreochromis nitoticus)

Nanomaterials mainly nanocomposites possess unique physical and chemical properties which makes them superior and indispensable. Though much research has been focused on the properties and application of nanocomposites, the eco-toxicity assessment is one among top priority, which aims to protect the population of concerned biological component and their ecosystem. With this objective, the present study has undertaken an initiation to evaluate the efficacy of chitosan-silver nanocomposite for methyl orange adsorption property (CS–AgNC) and also assessed the toxicity impact on growth parameters of freshwater Tilapia. Batch in vitro studies showed that all the tested dosages of the nanocomposite were effectively adsorbing maximum concentration of methyl orange. The synthesized nanocomposite was administrated to the tested fishes followed by the determination of various growth, nutritional parameters, gene expression of enzymatic antioxidants and liver, and intestinal tissues histology. Obtained results indicated that nanocomposite treatment was not projected as a toxic impact on all the tested growth, and nutritional parameters. Histology study showed that the exposure of Tilapia to nanocomposite has not shown any detrimental effect on antioxidants gene expression and liver, intestinal tissue architecture. Hence, all these findings indicated that chitosan-silver nanocomposite prepared in our present system was found to be biocompatible which suggested the possible utilization and release of the nanocomposite into the divergent ecosystem without affecting non-target organisms (NTO).

A potential herbal therapeutic for trichinellosis.

BackgroundTrichinellosis is a helminthic disease caused by Trichinella spiralis via the ingestion of raw or undercooked meat of infected animals. Current estimates indicate that 11 million humans have trichinellosis, worldwide. The effective use of anti-trichinella medications is limited by side effects and resistance which highlight the critical need for safe and effective drugs, particularly those derived from medicinal plants. Therefore, in the present study, we aimed to evaluate the efficacy of the ethanolic extract of Artemisia annua (A. annua) in treatment of experimentally induced trichinellosis.Materials and methodsTrichinellosis was induced experimentally in male 6–8 weeks BALB/c mice. BALB/c mice were divided into four groups, 10 mice each. One group was left uninfected and untreated, whereas three groups were infected with T. spiralis. One infected group of mice was left untreated (negative control) while the remaining two infected groups received either 300 mg/kg of the ethanolic extract of A. annua or 50 mg/kg of albendazole (positive control). All treatments started from the third day post-infection (dpi) for 3 successive days. All animals were sacrificed on the 7th dpi for evaluation of treatment efficacy.ResultsOur findings showed that A. annua treatment reduced the T. spiralis adult-worm count in the intestine of infected animals. Moreover, treatment with A. annua restored the normal intestinal architecture, reduced edema, alleviated inflammation as demonstrated by reduced inflammatory infiltrate and expression of TGF-β in intestinal tissues of A. annua-treated animals compared to infected untreated animals.ConclusionsOur findings show that A. annua extract is effective in treating experimentally induced trichinellosis which highlight the therapeutic potential of A. annua for intestinal trichinellosis.

Chitosan-Ocimum basilicum nanocomposite as a dietary additive in Oreochromis niloticus: Effects on immune-antioxidant response, head kidney gene expression, intestinal architecture, and growth

Several studies have looked into the use of basil, Ocimum basilicum (L.) in aquaculture as a dietary additive; however, more research is needed to see the possibility of it's including in nanocarriers in aquafeeds. An experiment was undertaken to highlight the efficacy chitosan-Ocimum basilicum nanocomposite (COBN), for the first time, on Nile tilapia (Oreochromis niloticus) growth, stress and antioxidant status, immune-related parameters, and gene expression. For 60 days, fish (average weight: 23.55 ± 0.08 g) were fed diets provided with different concentrations of COBN (g/kg): 0 g [COBN0], 1 g [COBN1], 2 g [COBN2], and 3 g [COBN3], where COBN0 was kept as control diet. Following the trial, the fish were challenged with pathogenic bacteria (Aeromonas sobria) and yeast (Candida albicans) infection. In comparison to the control (COBN0), a notable increase in growth parameters (weight gain, feed intake, and specific growth rate) and intestinal morphometric indices (average intestinal goblet cells count, villous width, and length) in all COBN groups was observed, where COBN2 and COBN3 groups had the highest values. The COBN diets significantly (p < 0.05) declined levels of serum triglycerides, glucose, cholesterol, and hepatic malondialdehyde. Moreover, the higher levels of serum biochemical biomarkers (growth hormone, total protein, globulin, and albumin), immunological parameters (phagocytic activity%, nitric oxide, and lysozyme), and hepatic antioxidant parameters (superoxide dismutase, total antioxidant capacity, and glutathione peroxidase) were obvious in the COBN2 and COBN3 groups followed by COBN1. The immune-antioxidant genes (TNF-α, IL-10, IL-1β, TGF-β, GPx, and SOD) were found to be considerably up-regulated in all COBN groups (COBN2 and COBN3 followed by COBN1). Fifteen days post-challenge with A. sobria and C. albicans, the highest survival rate was recorded in the COBN2 group (83.33 and 91.67%) followed by the COBN3 group (75 and 83.33%), respectively. The findings showed that a dietary intervention with COBN can promote growth, intestinal architecture, immunity, and antioxidant markers as well as protect O. niloticus against A. sobria and C. albicans infection. As a result, the COBN at a dose of 2 g/kg could be used as a food additive for the sustainable aquaculture industry.

Replacement of fishmeal with plant protein in the diets of juvenile lumpfish (Cyclopterus lumpus, L. 1758): Effects on digestive enzymes and microscopic structure of the digestive tract

This study investigated the effects of plant protein concentrates on the activity of digestive enzymes and microscopic structure of the intestine of juvenile lumpfish. The experiment was carried out using triplicate groups of lumpfish of 7 g average initial body weight, to 40 g average final weight, after 54 days. Four experimental diets were used; a fishmeal (FM) based control diet and three test diets with the plant protein concentrates; soy protein concentrate (SPC) and pea protein concentrate (PPC) (1:1 ratio) replacing FM in proportions of 25% (PP25), 50% (PP50) and 75% (PP75), respectively. Nested ANOVA showed several significant treatment differences in histomorphometry. Overall, fish fed test diets exhibited several changes such as shorter mucosal fold height (MFH) in the anterior intestine (AI), increased number of goblet cells (GCs) and width of lamina propria (WLP) in distal intestine (DI), compared to control. Principal component analysis (PCA) on histological indices showed that all three treatment groups (i.e. PP25, PP50 and PP75) had significantly altered overall intestinal architecture, compared to the control group. The observed enteritis was negatively related with the condition factor (CF). Regarding enzyme activities fewer changes were observed across diets. Fish fed the PP50 diet exhibited an increased activity of leucine aminopeptidase (LAP) in the pyloric caeca and a decrease on chymotrypsin (CHY) in the mid intestine (MI) compared to the control. Nevertheless, the overall changes captured by PCA on enzymes were associated only with condition factor (CF) and not with the diets. Overall, histological evaluation confirmed that lumpfish intestinal morphometry was significantly altered by plant protein ingredients at 25%, 50% and 75% levels, but replacing of FM up to 50% did not affected the growth and the enzyme activities up to 75% inclusion level.

Mechanism of Lysoforte in Improving Jejuna Morphology and Health in Broiler Chickens.

Lysoforte (LFT) plays a vital role in maintaining broilers' health and intestinal morphology. However, the mechanism behind the effects of LFT improving intestinal morphology and health is still unclear. Therefore, this study was implemented to explore the central genes linked to the regulatory effect of LFT. Seventy-five newly hatched Cobb 500 male broilers were randomly divided into three groups: control, LFT500, and LFT1000 groups, with 25 chicks per group. The control chicks were provided with the basal diet, and the birds in LFT500 and LFT1000 groups were offered the same basal diet with 500 g/ton and 1,000 g/ton LFT, respectively. GSE94622 dataset consisted of the control and two LFT-treated groups (LFT500 and LFT1000). Jejuna samples were obtained from Gene Expression Omnibus (GEO). Totally 106–344 DEGs were obtained by comparing LFT500 and LFT1000 vs. control and LFT1000 vs. LFT500. Gene ontology (GO) enrichment suggested that the DEGs are mainly related to the phosphatidylethanolamine biosynthetic process and neuron projection extension. KEGG analysis suggested the DEGs were enriched in AGE-RAGE, fatty acid elongation, ECM-receptor interaction (ECMRI), glycerophospholipid metabolism, focal adhesion, unsaturated fatty acids biosynthesis, and ABC transporters. Moreover, 29 genes, such as REG4, GJB1, KAT2A, APOA5, SERPINE2, ELOVL1, ABCC2, ANKRD9, CYP4V2, and PISD, might be closely related to promoting jejuna morphology in broilers. Taken together, our observation enhances the understanding of LFT in maintaining intestinal architecture and the general health of broiler chickens.

Paraprobiotics and Postbiotics of Lactobacillus delbrueckii CIDCA 133 Mitigate 5-FU-Induced Intestinal Inflammation.

Intestinal mucositis is a commonly reported side effect in oncology practice. Probiotics are considered an excellent alternative therapeutic approach to this debilitating condition; however, there are safety questions regarding the viable consumption of probiotics in clinical practice due to the risks of systemic infections, especially in immune-compromised patients. The use of heat-killed or cell-free supernatants derived from probiotic strains has been evaluated to minimize these adverse effects. Thus, this work evaluated the anti-inflammatory properties of paraprobiotics (heat-killed) and postbiotics (cell-free supernatant) of the probiotic Lactobacillus delbrueckii CIDCA 133 strain in a mouse model of 5-Fluorouracil drug-induced mucositis. Administration of paraprobiotics and postbiotics reduced the neutrophil cells infiltrating into the small intestinal mucosa and ameliorated the intestinal epithelium architecture damaged by 5-FU. These ameliorative effects were associated with a downregulation of inflammatory markers (Tlr2, Nfkb1, Il12, Il17a, Il1b, Tnf), and upregulation of immunoregulatory Il10 cytokine and the epithelial barrier markers Ocln, Cldn1, 2, 5, Hp and Muc2. Thus, heat-killed L. delbrueckii CIDCA 133 and supernatants derived from this strain were shown to be effective in reducing 5-FU-induced inflammatory damage, demonstrating them to be an alternative approach to the problems arising from the use of live beneficial microorganisms in clinical practice.

Patient-derived organoids for therapy personalization in inflammatory bowel diseases.

Inflammatory bowel diseases (IBDs) are chronic inflammatory disorders of the intestinal tract that have emerged as a growing problem in industrialized countries. Knowledge of IBD pathogenesis is still incomplete, and the most widely-accepted interpretation considers genetic factors, environmental stimuli, uncontrolled immune responses and altered intestinal microbiota composition as determinants of IBD, leading to dysfunction of the intestinal epithelial functions. In vitro models commonly used to study the intestinal barrier do not fully reflect the proper intestinal architecture. An important innovation is represented by organoids, 3D in vitro cell structures derived from stem cells that can self-organize into functional organ-specific structures. Organoids may be generated from induced pluripotent stem cells or adult intestinal stem cells of IBD patients and therefore retain their genetic and transcriptomic profile. These models are powerful pharmacological tools to better understand IBD pathogenesis, to study the mechanisms of action on the epithelial barrier of drugs already used in the treatment of IBD, and to evaluate novel target-directed molecules which could improve therapeutic strategies. The aim of this review is to illustrate the potential use of organoids for therapy personalization by focusing on the most significant advances in IBD research achieved through the use of adult stem cells-derived intestinal organoids.

Moringa oleifera Leaf Powder Dietary Inclusion Differentially Modulates the Antioxidant, Inflammatory, and Histopathological Responses of Normal and Aeromonas hydrophila-Infected Mono-Sex Nile Tilapia (Oreochromis niloticus).

This study aimed to detect the impact of Moringa oleifera leaf powder dietary inclusion on the antioxidant and innate immune responses of mono-sex Nile tilapia fingerlings. A total of 180 fingerlings were allocated in a random method into three groups with triplicate each. One group (1st group) received the control diet (basal diet (BD) free of moringa) and the other groups (2nd and 3rd) fed BD containing M. oleifera leaf powder at 5 and 10% of the diet, respectively. After 6 weeks of feeding, fish were randomly redistributed into four replicates and rested for 24 h. Then, each fish in the first two replicates was injected with 0.2 mL of PBS, while the others were injected with 0.2 mL of A. hydrophila suspension (1.8 × 106 CFU/mL). Healthy fish fed on M. oleifera leaf powder showed enhanced immune response manifested by significant increases in phagocytic and lysozyme activities with a marked H/L ratio (P < 0.05). In addition, significant alterations of the lymphocytic and heterophilic population in circulation with increasing infiltration in tissue such as the spleen were noticed. Also, M. oleifera significantly upregulated the antioxidants, CAT and GPx, proinflammatory cytokines, IL1-β, IL-8, and IFN-γ relative mRNA levels. On the other hand, following A. hydrophila challenging conditions, M. oleifera caused downregulations of IL1-β, IL-8, and IFN-γ transcription levels, and also lowered the CAT and GPx mRNA levels. In addition, a marked reduction of leukocytic infiltration plus a significant improvement of the degenerative changes in intestinal architecture has occurred. So, M. oleifera leaf powder can be included in the fish diet to enhance immune response under normal health conditions and lower the infection-associated inflammatory response.

Abstract 2651: The tankyrase inhibitor OM-153 demonstrates anti-tumor effect with a therapeutic index above 10

Abstract The catalytic enzymes tankyrase 1 and 2 (TNKS1/2) poly-ADP-ribosylate target proteins, including AXIN proteins, to earmark them for degradation by the ubiquitin-proteasomal system. Hence, inhibition of TNKS1/2 can stabilize AXIN proteins, and consequently β-catenin degradosomes, resulting in inhibition of WNT/β-catenin signaling. Although several potent small-molecules have been developed to inhibit TNKS1/2 and oncogenic WNT/β-catenin signaling, also showing convincing anti-tumor effects in numerous mouse cancer models, there are currently no TNKS1/2 inhibitors available in the clinic. The development has mainly been disadvantaged by concerns over previous reports unfolding intestinal toxicity, apparently caused by on-target and WNT/β-catenin signaling pathway-specific side effects, and a deficient therapeutic index. Here we show that the novel, highly potent and selective1,2,4-triazole-based TNKS1/2 inhibitor OM-153, displaying improved ADME-properties and mouse pharmacokinetics, reduced WNT/β-catenin signaling and tumor progression in COLO 320DM colon cancer xenografts upon peroral (PO) administration of 0.33-10 mg/kg twice daily (BID). In addition, combined OM-153 and anti-PD-1 treatment conferred a synergistic anti-tumor effect in a B16-F10 mouse melanoma model. In a 28-day repeated dose mouse toxicity study, body weight loss, intestinal damage and tubular damage in the kidney was documented after PO BID administration of 100 mg/kg. In contrast, in mice treated PO BID using 10 mg/kg, the intestinal architecture was intact and no atypical histopathological changes were observed in other organs, while clinical biochemistry and haematological analyses did not identify any changes indicating considerable toxicity. The results provide a scaffold for using OM-153 as a potential anti-cancer treatment and additional preclinical and clinical evaluations. Citation Format: Shoshy Alam Brinch, Enya Amundsen-Isaksen, Sandra Espada, Aleksandra Aizenshtadt, Lone Holmen, Clara Hammarström, Merete Høyem, Hanne Scholz, Gunnveig Grødeland, Sven T. Sowa, Albert Galera-Prat, Lari Lehtiö, Ilonka A.T.M. Meerts, Ruben G. G. Leenders, Anita Wegert, Stefan Krauss, Jo Waaler. The tankyrase inhibitor OM-153 demonstrates anti-tumor effect with a therapeutic index above 10 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2651.

Dietary fumonisin may compromise the nutritive value of feed and distort copper and zinc digestibility and retention in weaned piglets.

Fumonisins (FUM) have been reported to impede gut functioning in pigs. However, investigations into the possible effect on mineral metabolism are limited. Thus, the trial studied the apparent total tract digestibility (ATTD) and retention of dietary nitrogen and minerals, intestinal architecture, digestive enzymes activity and heat-shock protein 70 (Hsp70) activity. Eighteen weaned piglets of 7 weeks old were assigned to three groups and their feed either contained 0, 15 or 30 mg FUM/kg for 21 days. ATTD and retention of dietary N and minerals were measured in a 5- day long balance trial between Day 17 and Day 21. The digestible and metabolisable energy (DE and ME) content of the feeds were also determined. The body weights, cumulative feed intake, relative organ weights, digestive enzymes activity and intestinal morphology were not affected (p > 0.05) by dietary treatments. The DE content was significantly lower (p < 0.05) when the feed contained 15 mg/kg FUM, but no statistically reliable treatment effect was confirmed for ME content. Dietary FUM significantly lowered (p < 0.05) the ATTD of Ca and P but not (p > 0.05) N, K, Mg and Na. The relative retention rate of N, Ca, P, K, Mg and Na in all groups were not impacted (p > 0.05) by treatments. The ATTD and relative retention of Cu and Zn were remarkably (p < 0.05) lower in piglets fed FUM-contaminated feed. In addition, the expression of Hsp70 activity in the liver was significantly elevated (p < 0.05) in the highest treatment group. These findings suggest that a dietary dose of 15 or 30 mg FUM/kg diet distorts the nutritive value of the mixed feed, results in poor ATTD and retention rates of Zn and Cu, and elevate Hsp70 activity in the liver without altering intestinal architecture or digestive enzymes' activity in weaned piglets.

A β‐catenin‐TCF‐sensitive Locus Control Region Mediates GUCY2C Ligand Loss in Colorectal Cancer

BackgroundColorectal cancer most commonly arises from mutations in the tumor suppressor APC, or its downstream degradation target β‐catenin, initiating an oncogenic program of β‐catenin/TCF‐dependent nuclear transcription. Mechanisms coupling these mutations to tumorigenesis continue to be refined. The tumor suppressor axis regulated by the intestinal receptor guanylyl cyclase C (GUCY2C) and its paracrine ligands, guanylin and uroguanylin, contributes to intestinal homeostasis by regulating the proliferation, migration, and differentiation programs that maintain intestinal epithelial architecture. Furthermore, GUCY2C agonists oppose transformation in multiple mouse models of intestinal tumorigenesis. This signaling axis is among the earliest pathways silenced in tumorigenesis, reflecting an evolutionarily conserved pattern of GUCY2C retention, but ligand loss, in tumors. These observations suggest a pathophysiological model in which transformation orphans the receptor, silencing its signaling and lifting a block on tumorigenesis. Here, we examined the hypothesis that GUCY2C ligand loss arises from transcriptional silencing by β‐catenin/TCF.MethodsWe mapped the β‐catenin/TCF‐transcriptional program by RNA‐seq in four human colon cancer cell models. We then performed a comparative analysis of orthogonal approaches, including luciferase reporters, ChIP‐seq, CRISPR Cas9 knockout, and CRISPR epigenome editing to identify gene enhancers mediating GUCY2C ligand loss.ResultsRNA‐seq revealed a core β‐catenin/TCF‐transcriptional program of 1,289 genes, of which guanylin and uroguanylin were the seventh and twelfth most sensitive transcripts, reflecting transcriptional silencing. ChIP‐seq analyses of colon tissue revealed an evolutionarily conserved genomic locus encompassing the GUCY2C ligand genes, containing multiple regions of DNase hypersensitivity and H3K27ac enrichment, hallmarks of genetic enhancer loci. These markers were observed in normal colon, but not cancer tissue, consistent with enhancer inactivation and transcriptional silencing during transformation. ChIP‐seq in a colon cancer cell line revealed RNA Polymerase II and H3K27ac recruitment to the regions identified in human tissue, reflecting poised transcriptional machinery. Incorporation of the putative enhancer DNA into luciferase reporter constructs revealed a 2,683 bp locus control region (LCR) responsible for coordinated β‐catenin/TCF‐sensitive control of both GUCY2C ligand promoters. Both CRISPR‐Cas9 deletion of the LCR and epigenetic inactivation with a dCas9.KRAB repressor construct abolished β‐catenin/TCF‐sensitivity of GUCY2C ligand expression. Finally, LCR activation with a dCas9.VP64 transcriptional activator construct reconstituted GUCY2C ligand expression, overcoming gene silencing by β‐catenin/TCF.ConclusionsThis study reveals DNA elements regulating co‐repression of GUCY2C ligand transcription by β‐catenin/TCF, reflecting a novel step in tumorigenesis, and offers unique strategies to re‐establish hormone expression to oppose transformation.

Inflection in nutrient transporter genes leads to potential changes in small intestine histomorphology with improved nutrient retention in chicken under dietary synbiotic supplementation

This study investigated the role of synbiotic supplementation on gut histomorphology, expression of nutrient transporters, and nutrient utilization in broiler chicken. Bacitracin methylene disalicylate (BMD), Probiotic Lactobacillus acidophilus (LBA), and prebiotic Mannan-oligosaccharides (MOS) were used to formulate total 6 dietary treatments viz. T1 (control; basal diet), T2 (BMD @ 20 mg/kg diet), T3/T4 (0.1% MOS with 106 and 107 cfu LBA/g diet), and T5/T6 (0.2% MOS with 106 and 107 cfu LBA/g diet). A total of 288 day old chicken were allocated at random among six treatments, each with six replicates of eight chicken (48 birds/treatment). Results revealed higher villus height, crypt depth, villus height: crypt depth ratio, and histological surface magnification ratio values in chicken fed a combination of MOS (0.2%) and LBA (106 or 107 cfu/g diet). BMD supplemented birds had higher values of these parameters compared to control birds. The villus width, villus bottom area, and mucosal unit bottom area were all increased in birds supplemented with BMD, but were similar to birds fed a combination of MOS (0.1 percent) and LBA (107 cfu/g diet). The villus width, villus bottom area, and mucosal unit bottom area were higher in BMD supplemented birds which were similar to the birds fed a combination of MOS (0.1%) and LBA (107 cfu/g diet). The birds fed a combination of MOS (0.2%) and LBA (106 or 107 cfu/g diet) revealed upregulation of SGLT1 and GLUT5 expression in jejunum but no significant effect was observed on the expression of PePT1 and EAAT3 gene. The AME Diet, nitrogen, phosphorus, and calcium retentions were higher in birds fed a combination of MOS (0.2%) and LBA (106 or 107 cfu/g diet). However, the organic matter digestibility was higher in birds fed a combination of MOS (0.2%) and LBA (106 cfu/g diet) and BMD supplementation also improved the organic matter utilization compared to control. In conclusion, the supplementation of a combination of MOS (0.2%) and LBA (106 or 107 cfu/g diet) improves the intestinal architecture along with upregulation of SGLT1 and GLUT5 nutrient transporters and increase in nutrient digestibility in broiler chicken.

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.

Reversing Epithelial Polarity in Pluripotent Stem Cell-Derived Intestinal Organoids.

The inner surface of the intestine is a dynamic system, composed of a single layer of polarized epithelial cells. The development of intestinal organoids was a major breakthrough since they robustly recapitulate intestinal architecture, regional specification and cell composition in vitro. However, the cyst-like organization hinders direct access to the apical side of the epithelium, thus limiting their use in functional assays. For the first time, we show an intestinal organoid model from pluripotent stem cells with reversed polarity where the apical side faces the surrounding culture media and the basal side faces the lumen. These inside-out organoids preserve a distinct apico-basolateral orientation for a long period and differentiate into the major intestinal cell types. This novel model lays the foundation for developing new in vitro functional assays particularly targeting the apical surface of the epithelium and thus offers a new research tool to study nutrient/drug uptake, metabolism and host-microbiome/pathogen interactions.

Aging of intestinal stem cells.

SummaryThe intestine is one of the organs that relies on stem cell function for maintaining tissue homeostasis. Recent findings on intestinal aging show that intestinal architecture, such as villus length, crypt size, and cell composition changes in the aged crypts. The correspondent decline in the regenerative capacity of the intestine is mainly due to a decline in intestinal stem cell function upon aging, as the underlying mechanisms of aging intestinal stem cells are beginning to unravel. This review summarizes our current knowledge on stem cell-intrinsic mechanisms of aging of intestinal stem cells and their connection to extrinsic factors, such as niche cells and microbiota and will introduce recent approaches to attenuate or even revert the aging of intestinal stem cells.