MUC2 Gene Expression Research Articles

Effect of β-1/3,1/6-glucan upon immune responses and bacteria in the gut of healthy common carp (Cyprinus carpio).

β-glucans are frequently included in the diet of healthy common carp Cyprinus carpio as a pre-emptive measure for combatting disease. In order to study the effect this has on the relationship between the gut bacteria and host immune response, carp were maintained on either a β-glucan free diet or feed containing 0.1% MacroGard®, a β-1/3, 1/6-glucan, for up to 7 weeks and analysis of innate immune gene expression and molecular analysis of the gut bacteria was performed. The data reveals feeding of MacroGard® to healthy carp does not induce bactericidal innate immune gene expression in the gut but does appear to alter bacterial species richness that did not have a negative effect on overall health. Analysis of innate immune gene expression within the upper midgut revealed that there were significant changes over time in the expression of Interleukin (il)-1β, inducible nitric oxide synthase (inos), mucin (muc2) and C-reactive protein (crp2). Diet did not affect the number of copies of the bacterial 16s rDNA gene in the gut, used as a as a measure of total bacteria population size. However, PCR-denaturing gradient gel electrophoresis (DGGE) analysis revealed a shift in bacterial species richness with MacroGard feeding. Bactericidal immune gene expression of crp2, muc2 and il-1β was weakly correlated with gut bacteria population size indicating a potentially limited role of these genes in interacting with the gut bacteria in healthy carp in order to maintain gut homeostatic conditions. These findings highlight the importance of considering both host immunity and the microbiome together in order to fully elucidate the effeect of immunomodulants, such as β-glucans, upon gut health.

Pasteurella multocida specific bacteriophage suppresses P. multocida-induced inflammation: identification of genes related to bacteriophage signaling by Pasteurella multocida-infected swine nasal turbinate cells.

Although Pasteurella multocida is highly prevalent pathogen in animals and plays an important role in swine respiratory diseases, only a few studies on the use of bacteriophages specific to Pasteurella multocida disease have been reported. The object of this study was to investigate the therapeutic effect of specific P. multocida bacteriophages and to identify genes related to bacteriophage signaling utilizing RNA microarrays in swine nasal turbinate cells. Pas-MUP-1 phages were applied 24h prior to P. multocida infection (1 × 107cfu/ml) at several concentrations of bacterial infection. Cells were incubated to detect cytokines and 24h to detect mucin production. And real-time quantitative PCR was performed to examine related genes expression. To determine the change of total gene expression based on P. multocida and Pas-MUP-1 treatment, we performed RNA sequencing experiments. We found that P. multocida-infected PT-K75 cells show increased gene expression of IL-1β, IL-6, and Muc1 in a dose-dependent manner. Interestingly, these genes resulted in decreased expression in P. multocida pretreated with the P. multocida-specific Pas-MUP-1 bacteriophage. RNA sequencing analysis revealed that bacteriophage administration regulated genes associated with immune and inflammatory responses, and the regulated genes were dramatically concentrated in the cytokine/chemokine-based signaling pathways. Pas-MUP-1 treatment was shown to regulate P. multocida induced gene expression in the bacteria. These results suggest the specific bacteriophage has therapeutic potential as an alternative to antibiotic treatment to defend against P. multocida infection by altering inflammatory gene expression profiles.

The Antioxidant Alpha-Lipoic Acid Inhibits Proliferation and Invasion of Human Gastric Cancer Cells via Suppression of STAT3-Mediated MUC4 Gene Expression.

Background Metastasis and invasion are the main causes of mortality in gastric cancer. To improve the treatment of gastric cancer, the development of effective and innovative antitumor agents toward invasion and proliferation is needed. Alpha-lipoic acid (ALA), a naturally occurring thiol antioxidant, showed antiproliferative and cytotoxic effects on several cancers. So it is feasible to explore whether ALA can be used to inhibit proliferation and invasion in human gastric cancer. Methods The expression of MUC4 in human gastric cancer tissues was assayed by immunohistochemistry. Then, we performed in vitro cell proliferation and invasion analysis to explore the antitumor effect of ALA using AGS, BGC-823, and MKN-28 cells. To further explore the mechanism of ALA-mediated downregulation of MUC4, we cotransfected human gastric cancer cells with STAT3 siRNA and STAT3 overexpression construct. ChIP assays were carried out to find the relationship between MUC4 and STAT3. Results We found that the MUC4 gene was strongly expressed in human gastric cancer tissues. Meanwhile, ALA reduced proliferation and invasion of human gastric cancer cells by suppressing MUC4 expression. We also found that STAT3 was involved in the inhibition of MUC4 by ALA. Mechanistically, ALA suppressed MUC4 expression by inhibiting STAT3 binding to the MUC4 promoter region. Conclusion ALA inhibits both proliferation and invasion of gastric cancer cells by suppression of STAT3-mediated MUC4 gene expression.

Human Milk Oligosaccharides Differently Modulate Goblet Cells Under Homeostatic, Proinflammatory Conditions and ER Stress.

ScopeHuman milk oligosaccharides (hMOs) have beneficial effects on intestinal barrier function, but the mechanisms of action are not well understood. Here, the effects of hMOs on goblet cells, which indicate that some hMOs may enhance mucus barrier function through direct modulation of goblet cell function, are studied.Methods and resultsThe modulatory effects of 2′‐fucosyllactose (2′‐FL), 3‐fucosyllactose (3‐FL), lacto‐N‐triaose II (LNT2), and galacto‐oligosaccharides (GOS) on the expression of goblet cell secretory related genes MUC2, TFF3, and RETNLB, and the Golgi‐sulfotransferase genes CHST5 and GAL3ST2 of LS174T are determined by real‐time quantitative RT‐PCR. 3‐FL, LNT2, and GOS‐modulated LS174T gene expression profiles in a dose‐ and time‐dependent manner. In addition, the upregulation of MUC2 is confirmed by immunofluorescence staining. Effects of 2′‐FL, 3‐FL, LNT2, and GOS on gene transcription of LS174T are also assessed during exposure to TNF‐α, IL‐13, or tunicamycin. During TNF‐α challenge, 3‐FL and LNT2 enhance MUC2 and TFF3 gene expression. After IL‐13 exposure, 2′‐FL, 3‐FL, and LNT2 all show upregulating effects on MUC2; 3‐FL and LNT2 also enhance TFF3 expression. LNT2 significantly reverses Tm‐induced downregulation of TFF3, RETNLB, and CHST5.ConclusionThe findings indicate that hMOs may enhance mucus barrier function through direct modulation of intestinal goblet cells. Effects are structure‐ and stressor‐dependent.

Konjac glucomannan polysaccharide and inulin oligosaccharide enhance the colonic mucosal barrier function and modulate gut-associated lymphoid tissue immunity in C57BL/6J mice.

Both konjac glucomannan (KGM) and inulin oligosaccharide have been shown to improve bowel function, but their effects on the mucosal barrier function and immunity are not fully understood. The aim of the present study was to determine the effects of a low-level supplementation of dietary fibres on the colonic mucosal barrier function, antioxidant enzyme defence and immunity. C57BL/6J mice (6 weeks of age, eight per group) were randomly assigned to consume one of the following diets: control or control diet supplemented with 2 % (w/w) of KGM, inulin oligosaccharide (degree polymerisation = 8) or KGM+inulin (1 %, w/w each (K+I)). Fresh faeces were collected on days 19-21. Mice were killed on day 22 after fasting. Segments of colon tissues were processed for histological procedure and stained for acidic mucins and tight junction protein marker zona occludin-1 (ZO-1). The remaining tissues were processed to determine the gene expression of mucin 2, tight junction proteins, antioxidant enzymes and cytokines. The plasma cytokines were measured. Results indicated that KGM, inulin and K+I significantly increased the mucosal layer thickness, mucin density (granule number/crypt) and gene expression of Muc2 as compared with the control. All fibre treatments increased the gene expressions of ZO-1, occludin, glutathione peroxidase, glutathione S-transferase π, catalase and IL-10. In addition, all fibre treatments increased faecal butyrate and probiotics, and plasma IL-10 concentrations. In conclusion, supplementation of low-level, 2 % (w/w), of K+I was sufficient to enhance the mucosal barrier function and anti-inflammatory status.

MUC1 gene silencing inhibits proliferation, invasion, and migration while promoting apoptosis of oral squamous cell carcinoma cells.

The aim of the present study is to investigate the role of RNA interference in the inhibition of MUC1 gene expression in occurrence and metastasis of oral squamous cell carcinoma (OSCC) and its in-depth mechanisms. The OSCC and normal oral mucosa tissues, as well as normal oral epithelial cell line HOK and OSCC cell line SCC-4, Cal-27, TSCCA, Tca8113 were obtained to detect the expression of MUC1. Slug expression in OSCC and normal oral mucosa tissues was also determined. The OSCC cells were grouped to investigate the role of MUC1 gene silencing on proliferation, DNA replication, cell cycle distribution, apoptosis, colony formation ability, epithelial-mesenchymal transition (EMT), invasion, and migration of OSCC cells. We first found higher positive rate of MUC1 and Slug expression in OSCC tissues. Next, it was determined that higher expression of MUC1 was found in OSCC tissues and cells. Furthermore, silencing of MUC1 declined Slug expression, inhibited the proliferation, DNA replication, cell cycle progression, and EMT while inducing apoptosis of OSCC cells. Our study suggests that overexpression of MUC1 is found in OSCC, and MUC1 gene silencing could inhibit the proliferation, invasion, and migration while inducing apoptosis of OSCC cells.

In vitro-digested milk proteins: Evaluation of angiotensin-1-converting enzyme inhibitory and antioxidant activities, peptidomic profile, and mucin gene expression in HT29-MTX cells

Over the past decades, several studies investigated the health-promoting functions of milk peptides. However, to date many hurdles still exist regarding the widespread use of milk-derived bioactive peptides, as they may be degraded during gastrointestinal digestion. Thus, the aim of our study was to in vitro digest intact whey protein isolate (WPI) and casein proteins (CNP), mimicking in vivo digestion, to investigate their bioactive effects and to identify the potential peptides involved. Whey protein isolate and CNP were digested using a pepsin-pancreatin protocol and ultra-filtered (3-kDa cutoff membrane). A permeate (<3 kDa) and a retentate (>3 kDa) were obtained. Soy protein was included as a control (CTR). Angiotensin-1-converting enzyme inhibitory (ACE1-I) and antioxidant activity (AOX) were assessed and compared with those observed in undigested proteins and CTR. Furthermore, the permeate was characterized by nano-liquid chromatography electrospray ionization tandem mass spectrometry (LC-nano ESI MS/MS) using a shotgun peptidomic approach, and retentate was further digested with trypsin and analyzed by MS using a shotgun proteomic approach to identify potentially bioactive peptides. Further, the effects of WPI, CNP, and CTR retentate on cell metabolic activity and on mucus production (MUC5AC and MUC2 gene expression) were assessed in intestinal goblet HT29-MTX-E12 cells. Results showed that WPI permeate induced a significant ACE1-I inhibitory effect [49.2 ± 0.64% (SEM)] compared with undigested WPI, CNP permeate, and retentate or CTR permeate (10.40 ± 1.07%). A significant increase in AOX (1.58 ± 0.04 and 1.61 ± 0.02 µmol of trolox AOX equivalents per mg of protein, respectively) upon digestion was found in WPI. Potentially bioactive peptides associated with ACE1-I and antihypertensive effects were identified in WPI permeate and CNP retentate. At specific concentrations, WPI, CNP, and CTR retentate were able to stimulate metabolic activity in HT29-MTX-E12 cells. Expression of MUC5AC was increased by CNP retentate and unaltered by WPI retentate; MUC2 expression was significantly increased by 0.33 mg/g of CNP and reduced by 1.33 mg/g of CNP. Our results confirm that milk proteins may be rich sources of bioactive compounds, with the greatest beneficial potential of CNP at the intestinal goblet cell level.

Beta-glucan from Agrobacterium sp. ZX09 improves growth performance and intestinal function in weaned piglets.

Beta-glucan is currently under consideration as an alternative to in-feed antibiotics. The aim of the study was to investigate Agrobacterium sp. ZX09 beta-glucan on intestinal morphology, cytokine concentration, mucin expression and microbial populations of weaning piglets. Pigs were randomly assigned to one of five dietary treatments supplemented with 0, 25, 50, 100 and 200mg/kg beta-glucan. Data showed an increase in ADG at the 100mg/kg group (p=.03). A significant increase in villus height and reduction in crypt depth were fund in ileal tissue at the 100mg/kg inclusion level (p<.05). Dietary supplementation of 100mg/kg beta-glucan enhanced IL-10 concentration (p=.04) and gene expression of MUC1 and MUC2 (p<.05) in the jejunum. Dietary supplementation of 100mg/kg beta-glucan provoked the up-regulation of Lactobacillus counts and down-regulation of Escherichia coli counts in the caecum (p=.05). Data suggested that improved growth performance in response to beta-glucan supplementation at 100mg/kg in weaned piglets may be explained by the improved intestinal function.

Exopolysaccharides from Lactobacillus plantarum NCU116 Enhances Colonic Mucosal Homeostasis by Controlling Epithelial Cell Differentiation and c-Jun/Muc2 Signaling.

Probiotic lactobacilli and their exopolysaccharides (EPS) are thought to modulate mucosal homeostasis; however, their mechanisms remain elusive. Thus, we tried to clarify the role of exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) in the intestinal mucosal homeostasis. Our results indicated that EPS116 regulated the colon mucosal healing and homeostasis, enhanced the goblet cell differentiation, and promoted the expression of Muc2 gene in vivo and in vitro. Further experiments showed that EPS116 promoted the expression and phosphorylation of transcription factor c-Jun and facilitated its binding to the promoter of Muc2. Moreover, knocking down c-Jun or inhibiting its function in LS 174T cells treated with EPS116 led to decreased expression of Muc2, implying that EPS116 promoted the colonic mucosal homeostasis and Muc2 expression via c-Jun. Therefore, our study uncovered a novel model where EPS116 enhanced colon mucosal homeostasis by controlling the epithelial cell differentiation and c-Jun/Muc2 signaling.

The role of the gut microbiome in chronic liver disease: the clinical evidence revised.

SummaryRecent research has suggested a role for the intestinal microbiota in the pathogenesis and potential treatment of a wide range of liver diseases. The intestinal microbiota and bacterial products may contribute to the development of liver diseases through multiple mechanisms including increased intestinal permeability, chronic systemic inflammation, production of short-chain fatty acids and changes in metabolism. This suggests a potential role for pre-, pro- and synbiotic products in the prevention or treatment of some liver diseases. In addition, there is emerging evidence on the effects of faecal microbial transplant. Herein, we discuss the relationship between the intestinal microbiota and liver diseases, as well as reviewing intestinal microbiota-based treatment options that are currently being investigated.

131 Effects of Bacillus spp. probiotics on systemic immunity and intestinal health of weaned pigs experimentally infected with an enterotoxigenic Escherichia coli

Abstract The experiment was conducted to investigate the effects of Bacillus spp. on systemic immunity and intestinal health of weaned pigs infected with F18 E. coli. Weaned pigs (n = 36, 7.61 ± 0.40 kg BW) were randomly allotted to one of three treatments: a control diet and two diets supplemented with 500 mg/kg of Bacillus spp. strain 1 (PRO1) or strain 2 (PRO2). The experiment was conducted for 28 d, including 7 d before and 21 d after the first E. coli inoculation (d 0). The doses of F18 E. coli inoculum were 1010 cfu/3 mL oral dose daily for 3 consecutive days. Serum samples were collected on d 0 before inoculation, and d 3, 7, 14, and 21 post-inoculation (PI) to measure inflammatory meditators. All pigs were euthanized on d 21 PI to collect tissue samples for gut morphology, bacterial translocation, and gene expression analysis. Data were analyzed using the Mixed Procedure of SAS. Pigs in PRO1 had reduced (P &lt; 0.05) serum haptoglobin on d 3 and d 6 PI compared with pigs in control. Supplementation of PRO1 or PRO2 reduced (P &lt; 0.05) total coliforms in mesenteric lymph nodes on d 21 PI. Pigs in PRO1 had greater (P &lt; 0.05) ileal villi height than pigs in control. Pigs in PRO2 had greater (P &lt; 0.05) sulfomucin percentage in duodenal villi and greater (P &lt; 0.05) sialomucin percentage in jejunal villi than pigs in control. Supplementation of PRO1 also up-regulated (P &lt; 0.05) MUC2 gene expression in jejunal mucosa but reduced (P &lt; 0.05) COX2 and IL1B gene expression in ileal mucosa on d 21 PI, compared with control. In conclusion, supplementation of Bacillus spp. may reduce systemic inflammation and enhance intestinal health of weaned pigs infected with F18 E. coli.

Evaluation of mouse enteroids as a model for Lawsonia intracellularis infection

Lawsonia intracellularis, an obligate intracellular bacterium, is an important enteric pathogen in pig herds and horse farms worldwide. The hallmark feature of L. intracellularis infection is the proliferation of epithelial cells in intestinal crypts. A major limitation to the study of L. intracellularis infection is the lack of an in vitro model that reproduces the changes observed in proliferative enteropathy. Here we investigated the suitability of mouse enteroids as a model to study L. intracellularis infection. Mouse enteroids were microinjected with L. intracellularis, filter-sterilized L. intracellularis culture supernatant, or sterile cell culture media (DMEM). L. intracellularis antigen was detected in mouse enteroids by immunohistochemistry and was located mostly in the basal region of the epithelium. There was no differential growth of enteroids among treatment groups, and cellular proliferation was not increased in L. intracellularis-infected enteroids in relation to non-infected enteroids based on immunofluorescence staining. L. intracellularis infection did not induce changes in gene expression of Ki-67 (proliferation marker), Sox9 (marker for transit amplifying cells) and Muc2 (marker for goblet cells). These results indicate that although L. intracellularis antigen is detectable in mouse enteroids, indicating susceptibility to infection, mouse enteroids fail to replicate the cellular proliferation and gene expression changes observed in proliferative enteropathy. Nevertheless, we have successfully demonstrated that mouse enteroids can be used to model days-long intracellular pathogen infection, serving as potential models for the study of other pathogens of interest in veterinary medicine.

Fluorometholone modulates gene expression of ocular surface mucins.

Mucins are vital to keep the ocular surface hydrated. Genes encoding for mucins contain a glucocorticoid response element. The purpose of this study was to evaluate the effect of fluorometholone, a glucocorticoid receptor agonist used in the management of dry eye, on the gene expression of conjunctival and corneal epithelial cell mucins. Stratified cultures of human conjunctival and corneal epithelial cells were exposed to 25, 50 and 100nM of fluorometholone alone or in presence of mifepristone, a glucocorticoid receptor antagonist. The mRNA was isolated from the cells and reverse transcribed to cDNA. The cDNA was used for quantification of gene expression of mucin (MUC) 1, 4, 16 and 19 using real-time PCR. Fluorometholone caused a dose- and time-dependent increase in the gene expression of MUC1, MUC4, MUC16 and MUC19 in the conjunctival as well as corneal epithelial cells. Mifepristone, a glucocorticoid receptor antagonist, inhibited fluorometholone-mediated increase in the gene expression of conjunctival and corneal mucins. At the tested concentration, neither fluorometholone nor mifepristone caused any notable changes in the cellular phenotype or viability of conjunctival and corneal epithelial cells. Fluorometholone increases the gene expression of MUC1, MUC4, MUC16 and MUC19 in the conjunctival and corneal epithelial cells through activation of glucocorticoid receptors. The increased expression of mucins can be an additional possible mechanism contributing to the beneficial effects of fluorometholone in dry eye in addition to its well-known anti-inflammatory effects.

The influence of Farmatan®and Flimabend®on the mucosal immunity of broiler chicken

The aim of the present study was to monitor selected parameters of mucosal immunity in jejunum and ileum (immunoglobulin A [IgA], mucin 2 [MUC-2], and pro-inflammatory cytokines) in commercial broiler farm chicken after treatment with flubendazole (Flimabend®) and natural extract from chestnut wood (Farmatan®). A total of 24 forty-day-old Kalimero-Super Master hybrid chickens were divided into 4 groups (n = 6): the Fli group received Flimabend® per os, 100 mg/g suspension in 1.43 mg of active substance/kg body weight during 7 d of experiment; the Far group received Farmatan®per os at 0.2% concentration for 6 h/d during 5 d (experimental d 3 to 7); the Far + Fli group received a combination of doses administered in the same way as for the first two groups; and the C group represented control with no active substance administration. The concentrations of secretory IgA (sIgA) and MUC-2 and relative expression of selected immune parameters were evaluated. Our results show strong suppressive effect of the Farmatan® and Flimabend® combination on relative expression of IL-1β and IL-18 in selected parts of the intestine. On the other hand, administration of natural extract from selected chestnut wood (Farmatan®) increased expression of total IgA as well as concentration of sIgA in the studied parts of the chicken intestine. Moreover, expression and concentration of MUC-2 was positively affected by addition of Farmatan®. In contrast, 7-d administration of Flimabend® resulted in upregulation of pro-inflammatory cytokines and decrease in IgA and MUC-2 gene expression. In conclusion, for maintenance of mucosal immunity via activation of IgA and mucin production, the long-term preventive use of Farmatan® is a suitable choice.

Cervical mucus sialic acid content determines the ability of frozen-thawed ram sperm to migrate through the cervix.

The aim of this study was to investigate the properties and to functionally characterize the cervical mucus that modulates sperm transport through the cervix by using ewe breeds with a divergent pregnancy rate (Belclare and Suffolk; high and low, respectively) following cervical insemination using frozen-thawed semen. Sperm number, as well as sialic acid and fucose content in both the channels and in the lumen of different regions of the cervix were quantified in inseminated Belclare and Suffolk ewes. Expression of glycosyltransferase and MUC genes, glycosidase activity and sialic acid speciation in follicular phase cervical tissue and mucus were assessed. More spermatozoa were found in the cervical channels in the region closest to the cervical os in Belclare than Suffolk ewes (P < 0.05) and Suffolk ewes had a higher sialic acid content in the cervical channels than Belclare ewes (P < 0.05) in all regions of cervix. Suffolk ewes had significantly higher expression of FUT1, ST6GAL1 and MUC5AC than Belclare ewes. There was no difference between the breeds in glycosidase activity (P > 0.05). Levels of Neu5Ac were higher in Belclare than Suffolk ewes (P < 0.05) and levels of Neu5Gc was higher in Suffolk than Belclare ewes (P < 0.05). Competitive sperm penetration assays demonstrated that frozen-thawed sperm progression increased when cervical mucus was incubated with sialyllactose prior to a sperm penetration test (P < 0.05). These results suggest that the difference between Belclare and Suffolk ewes in sperm transport with frozen-thawed semen is due to the higher concentration of sialic acid within channels, which binds to spermatozoa and reduces their ability to traverse the cervix.

Autolyzed Yeast (Saccharomyces cerevisiae) Supplementation Improves Performance While Modulating the Intestinal Immune-System and Microbiology of Broiler Chickens

The objective of this study was to evaluate the effects of dietary autolyzed yeast (AY; Saccharomyces cerevisiae) supplementation on growth performance, immune system, and intestinal bacterial count in broiler chickens. A total of 1,260 one-day-old male Ross AP95 chicks were placed in a completely randomized design (4 treatments, 7 replicates each, and 45 birds/replicate). The treatments were: basal diet – negative control (NC); basal diet supplemented with 55 ppm of zinc bacitracin – positive control (PC); NC + 0.2% of AY; NC + 0.4% of AY. The diets were formulated based on corn-soybean meal with 5% inclusion of wheat bran and 5% of poultry by-product meal. At 7 days of age, all birds were eye drop-vaccinated with live vaccine against coccidiosis. At 8 and 21 d of age, one chicken per pen was then euthanized by cervical dislocation to collect ileal and cecal contents for enumeration of Enterococcus sp., Escherichia coli and Lactobacillus sp. Yet, ileal samples were collected to analyze the gene expression of Claudin-1, IL-1β, IL-4, TLR-4 and MUC-2 through real time PCR. On d 21, it was observed that the inclusion of 0.2% of AY improved FCR (P < 0.05) when compared to the NC treatment. In the overall experimental period, the inclusion of zinc bacitracin and 0.4% of AY improved FCR (P < 0.05) compared to the NC group. On d 8, supplementation of 0.2 % of AY increased Enterococcus and both concentrations of AY reduced Lactobacillus in the ileal digesta compared to birds supplemented with zinc bacitracin. On d 21, 0.2% of AY reduced E. coli in the cecal digesta. On d 8, AY supplementation downregulated the expression of TLR-4 vs. the PC group (P = 0.04). On d 21, supplementation of AY upregulated the expression of IL-1β (P < 0.05) vs. the NC group. Supplementation of AY improved the growth performance of broiler chickens vaccinated against coccidiosis, partially explained by the modulation of the intestinal microbiota and immune-system.

Bordetella bronchiseptica bateriophage suppresses B. bronchiseptica-induced inflammation in swine nasal turbinate cells.

The development of therapeutic bacteriophages will provide several benefits based on an understanding the basic physiological dynamics of phage and bacteria interactions for therapeutic use in light of the results of antibiotic abuse. However, studies on bacteriophage therapeutics against microbes are very limited, because of lack of phage stability and an incomplete understanding of the physiological intracellular mechanisms of phage. The major objective of this investigation was to provide opportunity for development of a novel therapeutic treatment to control respiratory diseases in swine. The cytokine array system was used to identify the secreted cytokines/chemokines after Bordetella bronchiseptica infection into swine nasal turbinate cells (PT-K75). We also performed the real-time quantitative PCR method to investigate the gene expression regulated by B. bronchiseptica infection or bacteriophage treatment. We found that B. bronchiseptica infection of PT-K75 induces secretion of many cytokines/chemokines to regulate airway inflammation. Of them, secretion and expression of IL-1β and IL-6 are increased in a dose-dependent manner. Interestingly, membrane-bound mucin production via expression of the Muc1 gene is increased in B. bronchiseptica-infected PT-K75 cells. However, cytokine production and Muc1 gene expression are dramatically inhibited by treatment with a specific B. bronchiseptica bacteriophage (Bor-BRP-1). The regulation of cytokine profiles in B. bronchiseptica-induced inflammation by B. bronchiseptica bacteriophage is essential for avoiding inappropriate inflammatory responses. The ability of bacteriophages to downregulate the immune response by inhibiting bacterial infection emphasizes the possibility of bacteriophage-based therapies as a novel anti-inflammatory therapeutic strategy in swine respiratory tracts.

Ingestion of paddy rice increases intestinal mucin secretion and goblet cell number and prevents dextran sodium sulfate-induced intestinal barrier defect in chickens.

Paddy rice is a potential feed grain for chickens, whose strong gizzards can crush the hull. Here, we investigated whether paddy rice rich in hull-derived water-insoluble dietary fiber stimulates intestinal mucin secretion and production, as well as the possible involvement of paddy rice in intestinal barrier function. Layer male chicks at 7 d of age were divided into four groups according to the diet: corn, polished rice, brown rice, or paddy rice (650g/kg diet), which they ate for 14 consecutive days. At 21 d of age, the birds were refed their experimental diets, and small intestinal mucin fractions were collected to determine intestinal mucin content. Small intestinal mucin secretion was induced most strongly in the paddy rice group (Experiment 1). The rank order of diet-induced mucin secretion was paddy rice > corn = brown rice > polished rice. Ileal MUC2 gene expression and ileal number of goblet cells were highest in the paddy rice group (Experiment 1). A study of bromodeoxy-U uptake into ileal epithelial cells indicated the increase in goblet cells in the paddy rice group was related to accelerate epithelial cell migration (Experiment 2). A single supplementation of isolated rice hulls without kernels increased MUC2 gene expression and goblet cell numbers (Experiment 3), suggesting the importance of the hull's bulk-forming capacity on mucin production. Finally, chicks fed corn or paddy rice were orally administered dextran sodium sulfate (DSS) to disrupt intestinal barrier function. In the DSS-treated birds, the intestinal permeability of fluorescein isothiocyanate dextran in the everted gut sacs was much lower in the paddy rice group than in the corn group (Experiment 4), showing that paddy rice protects against mucosal disruption. In conclusion, ingestion of paddy rice increases intestinal mucin secretion and production through enhanced MUC2 gene expression and epithelial turnover and prevents DSS-induced intestinal barrier defects in chickens.

Oral consumption of cinnamon enhances the expression of immunity and lipid absorption genes in the small intestinal epithelium and alters the gut microbiota in normal mice

Because cinnamon is orally ingested, its biological activity is likely to influence the small intestinal epithelium and the microbiota along the intestinal tract. We investigated small intestinal epithelial gene expression associated with immunity and lipid metabolism and measured IgA level in the small intestines and sera of mice fed cinnamon. We also analyzed microbial changes in the small and large intestines and feces using pyrosequencing of the 16 s rRNA gene. Cinnamon increased the gene expression of Muc2, RegIIIγ, and Pigr, which are necessary for defense against gut bacteria. It also increased the gene expression of GATA4 and Slc27a2, which are implicated in lipid absorption. Cinnamon increased luminal IgA level but not serum IgA and decreased the class Gammaproteobacteria (classified within the phylum Proteobacteria), particularly the genus Pseudomonas, in the large intestine. Cinnamon thus affected immunity and lipid absorption in the small intestinal epithelium and microbial composition in the intestines.

The protective role of glutamine on enteropathy induced by high dose of soybean meal in turbot, Scophthalmus maximus L.

The aim of this study is to investigate the protective effect and regulatory mechanism of Gln on the enteropathy of turbot induced by soybean meal. Three isonitrogenous and isolipidic practical diets were formulated with fish meal basis (FM), soy protein replacing 40% fish meal protein (SBM) and dietary 2% Ala-Gln in SBM (Ala-Gln) for sextuple replicates of 30 fish each tank for 12 weeks. The fish in SBM group presented distinct enteropathy including declined absorptive surface and overt infiltration of mixed leukocytes in the lamina propria. However, the turbot fed Ala-Gln diet showed the integrity of the intestinal tissue structure and higher ADC and AKP levels like fish in FM group. Compared to SBM group, the inclusion of 2% Ala-Gln in SBM-basal diet significantly elevated the expression of barrier-forming tight junction proteins claudin-4, occludin and ZO-1 and depressed the expression of pro-inflammatory cytokines TNF-α in the intestine. Fish fed Ala-Gln diet showed remarkable increased MUC-2 and PPAR-γ gene expression as well as reduced expression of NF-κB and MLCK equivalent to the level of FM group. Additionally, Gln significantly elevated the relative abundance of Proteobacteria (Vibrio spp.) and declined Bacteroidetes (Bacteroides spp.) relative abundance. The similarity of intestinal microbial communities in Ala-Gln group was notably closer to FM group from PCoA, UPGMA and Heatmap analyses based on weighted UniFrac distance. In conclusion, dietary Gln could probably enhance intestinal barrier function to alleviate enteropathy by improving MUC-2 and PPAR-γ expression and inhibiting NF-κB-MLCK signaling pathway, as well as altering microbiota.