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Mucociliary clearance is an important phenomenon inside the respiratory system as a first defensive mechanism against pathogens. Therefore, any assumption considered for the mucociliary clearance and affects its functionality must be validated. The present research deals with the effects of boundary conditions on the movement of upper-convected Maxwell and high viscosity Newtonian mucus layers, numerically. Furthermore, the validity of replacing the viscoelastic mucus layer with a high viscosity Newtonian layer is evaluated. The airway surface liquid layer is considered a two-layer model including non-Newtonian mucus and Newtonian periciliary layers. Four cyclic boundary conditions are imposed at the mucus-periciliary interface as the cilia movement to obtain variations of mucociliary clearance. The upper boundary of the mucus layer is also exposed to different shear stress levels including free slip, cough, and sneeze conditions. By investigation of velocity variations inside mucus and periciliary layers, it is concluded the differences between viscoelastic and Newtonian mucus are not negligible. The maximum velocity differences between the two fluids are more than 52% and 215% during cough and sneeze, respectively. The results show there is a high order of dependency between the relaxation time and the imposed boundary conditions at the mucus-periciliary interface that leads to the invalidation of replacing two fluids with each other. Moreover, the results show substituting the viscoelastic mucus with a high viscosity Newtonian one depends on the mucus-periciliary interface boundary condition. If an independent time-varying boundary condition is used, the substitution leads to an error less than 7% under different shear stress levels. However, time-varying boundary condition shows 38% and 88% differences between high viscosity Newtonian and viscoelastic mucus layers. Furthermore, neglecting the recovery stroke leads to a velocity underestimation up to 50% by substituting viscoelastic mucus with a high viscosity Newtonian one. Therefore, replacing the viscoelastic mucus with a high viscosity Newtonian one is not acceptable for numerical simulations.

Mucus layer movement inside the airway system is an important phenomenon as the first defensive mechanism against pathogens. This research deals with the mucus velocity variations inside the nasal cavity using two different power law and thixotropic mucus layers. The cilia movement is replaced with four cyclic velocity profiles at the lower boundary of the mucus layer, while the upper boundary is exposed to the free-slip condition. The effects of boundary conditions and different fluid parameters are evaluated on the mucus flow. Furthermore, the replacement of power law and thixotropic mucus layers with a high viscous Newtonian mucus is examined under the free-slip condition at the mucus upper boundary. The adaptation rate is used as the criteria for replacing fluids instead of each other. The results show the mucus flow has enough time to adjust the changes from the lower boundary and the recovery stroke does not affect the mucus velocity in the effective stroke. Moreover, it is observed that the mucus flow variations are the same under the influence of recovery, breakdown, and breakdown exponent parameters. However, the effects of the exponent parameter on the mucus flow are more than the other two parameters in the recovery stroke. It is concluded that the assumption for replacing the power law mucus with a high viscous Newtonian one is acceptable. However, this assumption leads to the maximum error of 98.5% for thixotropic mucus in the recovery stroke.

Firefighters and workers in some industries often inhale (polluted) air at high temperatures, eg, 50–200°C, and very low relative humidity, say, 5–10%. Such severe environmental conditions may cause health problems or further aggravate existing ones. As in vivo tests are very complex, potentially harmful and costly, and in vitro experiments often lack high resolution and predictive capability, in silico studies become a priority. They may include computer simulations of the air-particle dynamics in human lung-airway models to understand the convection heat transfer as well as pollutant transport, deposition and uptake at high inlet temperatures and low relative humidity. In this study, detailed modeling, simulation and analysis focuses on two-phase flow in a human upper lung-airway model with a realistic 3D mucus lining. Especially modeling of the upper airway mucus layers was of interest in order to simulate the vapor mass transfer to the airflow because of its primary function for airway humidification. Naturally, the humidification causes water loss in the mucus layer, which leads to its reduction or even depletion when exposed to relatively high temperatures. Accounting for the changes in thickness and the rise in temperature in the mucus layer allows for the determination of locations of thermal injury in the human airways due to continuous exposure of such abnormal inhalation conditions. Different temperature profiles and local changes in mucus layer thickness were studied for ranges of severe inlet temperature conditions at a representative flow rate of 20 LPM (liters per minute). For inlet temperature reaching 100°C, mucus-layer thinning was observed in the upper airways. Interestingly, as a confirmation of the Reynolds analogy, the areas of significant wall heat flux and associated wall shear stress coincided with the regions of highest mucus evaporation, resulting in the humidification of the air with low relative humidity. Model development and mucus layer generation were done using C++ programming. All computer simulations were carried out using the open-source computational fluid dynamics toolbox OpenFOAM.

BackgroundAn interaction of the food types with the gut microbiota changes is deeply implicated in human health and disease. To verify whether animal-based diets would lead to gut dysbiosis, systemic inflammation and inflammatory pathogenesis, we fed mice with chondroitin sulfate (CS), a sulfate-containing O-glycan naturally occurring in livestock and poultry products, and monitored the dynamic changes of microbial flores, inflammatory signatures, and pathogenic hallmarks.ResultsA metagenomic gut microbiota analysis revealed the overgrowth of sulfatase-secreting bacteria and sulfate-reducing bacteria in the gastrointestinal tracts of mice upon daily CS feeding. Sulfatase-secreting bacteria compromise gut integrity through prompting mucin degradation and mucus lesions, which were evident from the upregulation of secretary leukocyte protease inhibitor (SLPI) and mucin 1/4 (MUC-1/4). A synchronous elevation of lipopolysaccharide (LPS) and tumor necrosis factor α (TNF-α) levels in the serum as well as cerebral, hepatic, cardiac and muscular tissues suggests bacterial endotoxinemia, chronic low-grade inflammation and mitochondrial dysfunction, eventually leading to the onset of global inflammatory pathogenesis towards arthritis, dementia, tumor, and fatty liver.ConclusionsCS triggers the early-phase and multi-systemic pathogenesis like arthritis, dementia, tumor, and fatty liver by enhancing gut opportunistic infection and evoking low-grade inflammation in mice. A plausible reason for the inconsistency of CS in treatment of osteoarthritis (OA) was also discussed.

The colon epithelium is protected from the luminal microbes as recently revealed by an inner firmly attached mucus layer impervious to bacteria and an outer loose mucus layer that is the habitat of bacteria. For an additional understanding of these layers, we analyzed the protein composition of these two mucus layers from the mouse colon. Proteomics using nano-LC-MS and MS/MS revealed more than 1000 protein entries. As the mucus layers contain detached cells, a majority of the proteins had an intracellular origin. However, at least 44 entries were described as secreted proteins and predicted to be mucus constituents together with extracellular/plasma and bacterial proteins, the latter largely in the loose mucus layer. A major protein was the Muc2 mucin that by its net-like disulfide-bonded polymer structure builds the mucus. When guanidinium chloride insoluble Muc2 units were analyzed, N-terminal parts of the Fc-gamma binding protein (Fcgbp) was found to be covalently attached in mouse and human colon, whereas its C-terminus was lost by reducing the disulfide bonds. In conclusion, the Fcgbp protein is probably cleaved at GD/PH and covalently attached to Muc2 via one or several of its von Willebrand D domains.

Cathepsin K has been shown to exhibit antimicrobial and anti-inflammatory activities in the mouse colon. To further elucidate its role, we used Ctsk -/- mice and demonstrated that the absence of cathepsin K was accompanied by elevated protein levels of related cysteine cathepsins (cathepsins B, L, and X) in the colon. In principle, such changes could result in altered subcellular localization; however, the trafficking of cysteine cathepsins was not affected in the colon of Ctsk -/- mice. However, cathepsin K deficiency affected the extracellular matrix constituents, as higher amounts of collagen IV and laminin were observed. Moreover, the localization pattern of the intercellular junction proteins E-cadherin and occludin was altered in the colon of Ctsk -/- mice, suggesting potential impairment of the barrier function. Thus, we used an ex vivo method for assessing the mucus layers and showed that the absence of cathepsin K had no influence on mucus organization and growth. The data of this study support the notion that cathepsin K contributes to intestinal homeostasis and tissue architecture, but the lack of cathepsin K activity is not expected to affect the mucus-depending barrier functions of the mouse colon. These results are important with regard to oral administration of cathepsin K inhibitors that are currently under investigation in clinical trials.

Elasmobranchs represent a distinct group of cartilaginous fishes that harbor a remarkable ability to heal wounds rapidly and without infection. To date very little work has addressed this phenomenon although it is suggested that antibiotic capabilities associated with epidermal surfaces may be a factor. The study of benefits derived from mutualistic interactions between unicellular and multicellular organisms is a rapidly growing area of research. Here we survey and identify bacterial associates of three ray and one skate species in order to assess the potential for antibiotic production from elasmobranch associated bacteria as a novel source for new antibiotics.

We have recently shown that the colon is protected by an inner mucus layer that efficiently separates the bacteria in the outer mucus from the epithelial cells. The inner mucus is impervious for bacteria and built by a network formed by the MUC2 mucin. Lack or defects in this inner mucus layer allow bacteria to reach the epithelia, something that triggers colon inflammation.

We normally live in symbiosis with ∼10 13 bacteria present in the colon. Among the several mechanisms maintaining the bacteria/host balance, there is limited understanding of the structure, function, and properties of intestinal mucus. We now demonstrate that the mouse colonic mucus consists of two layers extending 150 μm above the epithelial cells. Proteomics revealed that both of these layers have similar protein composition, with the large gel-forming mucin Muc2 as the major structural component. The inner layer is densely packed, firmly attached to the epithelium, and devoid of bacteria. In contrast, the outer layer is movable, has an expanded volume due to proteolytic cleavages of the Muc2 mucin, and is colonized by bacteria. Muc2 −/− mice have bacteria in direct contact with the epithelial cells and far down in the crypts, explaining the inflammation and cancer development observed in these animals. These findings show that the Muc2 mucin can build a mucus barrier that separates bacteria from the colon epithelia and suggest that defects in this mucus can cause colon inflammation.

SummaryThe intestinal mucus layer provides a barrier limiting bacterial contact with the underlying epithelium. Mucus structure is shaped by intestinal location and the microbiota. To understand how commensals modulate gut mucus, we examined mucus properties under germ-free (GF) conditions and during microbial colonization. Although the colon mucus organization of GF mice was similar to that of conventionally raised (Convr) mice, the GF inner mucus layer was penetrable to bacteria-sized beads. During colonization, in which GF mice were gavaged with Convr microbiota, the small intestine mucus required 5 weeks to be normally detached and colonic inner mucus 6 weeks to become impenetrable. The composition of the small intestinal microbiota during colonization was similar to Convr donors until 3 weeks, when Bacteroides increased, Firmicutes decreased, and segmented filamentous bacteria became undetectable. These findings highlight the dynamics of mucus layer development and indicate that studies of mature microbe-mucus interactions should be conducted weeks after colonization.

The intestinal epithelium is covered with mucus with the main structural building block being the densely O-glycosylated MUC2 mucin. The intestinal epithelium is exposed to ingested material, our digestive machinery, and large amounts of microorganisms. Mucus is the first line of defense and aids to limit exposure to all these threats to the epithelium. In the small intestine, mucus acts as a matrix, which contains antimicrobial products, such as defensins and immunoglobulin A that limit epithelial exposure to the luminal bacteria. In the colon, the stratified inner mucus layer acts as a physical barrier excluding bacteria from the epithelium. Bacterial penetration of this normally restricted zone is observed in many colitis models and also in patients with ulcerative colitis. Mucus defects that allow bacteria to reach the epithelium and to stimulate an immune system response can lead to the development of intestinal inflammation. The current state of our knowledge concerning the function of the mucus layers and the main mucin component, MUC2, in inflammatory bowel disease is described in this review.

The enormous bacterial load and mechanical forces in colon create a special requirement for protection of the epithelium. In the distal colon, this problem is largely solved by separation of the bacteria from the epithelium by a firmly attached inner mucus layer. In addition, an outer mucus layer entraps bacteria to be cleared by distal transport. The mucus layers contain a network of Muc2 mucins as the main structural component. Here, the renewal rate of the inner protective mucus layer was studied as well as the production and secretion of Muc2 mucin in the distal colon. This was performed by intraperitoneal injection of N-azidoacetyl-galactosamine (GalNAz) that was in vivo incorporated during biosynthesis of O-glycosylated glycoproteins. The only gel-forming mucin produced in the colon is the Muc2 mucin and as it carries numerous O-glycans, the granulae of the goblet cells producing Muc2 mucin were intensely stained. The GalNAz-labeled glycoproteins were first observed in the Golgi apparatus of most cells. Goblet cells in the luminal surface epithelium had the fastest biosynthesis of Muc2 and secreted material already three hours after labeling. This secreted GalNAz-labeled Muc2 mucin formed the inner mucus layer. The goblet cells along the crypt epithelium accumulated labeled mucin vesicles for a longer period and secretion of labeled Muc2 mucin was first observed after 6 to 8 h. This study reveals a fast turnover (1 h) of the inner mucus layer in the distal colon mediated by goblet cells of the luminal surface epithelium.

The mucus layer continuously covering the gastric mucosa consists of a loosely adherent layer that can be easily removed by suction, leaving a firmly adherent mucus layer attached to the epithelium. These two layers exhibit different gastroprotective roles; therefore, individual regulation of thickness and mucin composition were studied. Mucus thickness was measured in vivo with micropipettes in anesthetized mice [isoflurane; C57BL/6, Muc1−/−, inducible nitric oxide synthase (iNOS)−/−, and neuronal NOS (nNOS)−/−] and rats (inactin) after surgical exposure of the gastric mucosa. The two mucus layers covering the gastric mucosa were differently regulated. Luminal administration of PGE2increased the thickness of both layers, whereas luminal NO stimulated only firmly adherent mucus accumulation. A new gastroprotective role for iNOS was indicated since iNOS-deficient mice had thinner firmly adherent mucus layers and a lower mucus accumulation rate, whereas nNOS did not appear to be involved in mucus secretion. Downregulation of gastric mucus accumulation was observed in Muc1−/− mice. Both the firmly and loosely adherent mucus layers consisted of Muc5ac mucins. In conclusion, this study showed that, even though both the two mucus layers covering the gastric mucosa consist of Muc5ac, they are differently regulated by luminal PGE2and NO. A new gastroprotective role for iNOS was indicated since iNOS−/− mice had a thinner firmly adherent mucus layer. In addition, a regulatory role of Muc1 was demonstrated since downregulation of gastric mucus accumulation was observed in Muc1−/− mice.

Colon has been shown to have a two-layered mucus system where the inner layer is devoid of bacteria. However, a complete overview of the mouse gastrointestinal mucus system is lacking. We now characterize mucus release, thickness, growth over time, adhesive properties, and penetrability to fluorescent beads from stomach to distal colon. Colon displayed spontaneous mucus release and all regions released mucus in response to carbachol and PGE2, except the distal colon and domes of Peyer's patches. Stomach and colon had an inner mucus layer that was adherent to the epithelium. In contrast, the small intestine and Peyer's patches had a single mucus layer that was easily aspirated. The inner mucus layer of the distal colon was not penetrable to beads the size of bacteria and the inner layer of the proximal colon was only partly penetrable. In contrast, the inner mucus layer of stomach was fully penetrable, as was the small intestinal mucus. This suggests a functional organization of the intestinal mucus system, where the small intestine has loose and penetrable mucus that may allow easy penetration of nutrients, in contrast to the stomach, where the mucus provides physical protection, and the colon, where the mucus separates bacteria from the epithelium. This knowledge of the mucus system and its organization improves our understanding of the gastrointestinal tract physiology.

The normal intestinal microbiota inhabits the colon mucus without triggering an inflammatory response. The reason for this and how the intestinal mucus of the colon is organized have begun to be unraveled. The mucus is organized in two layers: an inner, stratified mucus layer that is firmly adherent to the epithelial cells and approximately 50 μm thick; and an outer, nonattached layer that is usually approximately 100 μm thick as measured in mouse. These mucus layers are organized around the highly glycosylated MUC2 mucin, forming a large, net-like polymer that is secreted by the goblet cells. The inner mucus layer is dense and does not allow bacteria to penetrate, thus keeping the epithelial cell surface free from bacteria. The inner mucus layer is converted into the outer layer, which is the habitat of the commensal flora. The outer mucus layer has an expanded volume due to proteolytic activities provided by the host but probably also caused by commensal bacterial proteases and glycosidases. The numerous O -glycans on the MUC2 mucin not only serve as nutrients for the bacteria but also as attachment sites and, as such, probably contribute to the selection of the species-specific colon flora. This observation that normal human individuals carry a uniform MUC2 mucin glycan array in colon may indicate such a specific selection.

Although the detection of viable probiotic bacteria following their ingestion and passage through the gastrointestinal tract (GIT) has been well documented, their mucosal attachment in vivo is more difficult to assess. In this study, we investigated the survival and mucosal attachment of multi-strain probiotics transiting the rat GIT. Rats were administered a commercial mixture of the intestinal probiotics Lactobacillus acidophilus LA742, Lactobacillus rhamnosus L2H and Bifidobacterium lactis HN019 and the oral probiotic Streptococcus salivarius K12 every 12 h for 3 days. Intestinal contents, mucus and faeces were tested 6 h, 3 days and 7 days after the last dose by strain-specific enumeration on selective media and by denaturing gradient gel electrophoresis. At 6 h, viable cells and DNA corresponding to all four probiotics were detected in the faeces and in both the lumen contents and mucus layers of the ileum and colon. Viable probiotic cells of B. lactis and L. rhamnosus were detected for 7 days and L. acidophilus for 3 days after the last dose. B. lactis and L. rhamnosus persisted in the ileal mucus and colon contents, whereas the retention of L. acidophilus appeared to be relatively higher in colonic mucus. No viable cells of S. salivarius K12 were detected in any of the samples at either day 3 or 7. The study demonstrates that probiotic strains of intestinal origin but not of oral origin exhibit temporary colonisation of the rat GIT and that these strains may have differing relative affinities for colonic and ileal mucosa.

In discussions on intestinal protection, the protective capacity of mucus has not been very much considered. The progress in the last years in understanding the molecular nature of mucins, the main building blocks of mucus, has, however, changed this. The intestinal enterocytes have their apical surfaces covered by transmembrane mucins and the whole intestinal surface is further covered by mucus, built around the gel-forming mucin MUC2. The mucus of the small intestine has only one layer, whereas the large intestine has a two-layered mucus where the inner, attached layer has a protective function for the intestine, as it is impermeable to the luminal bacteria.

Mucus thickness is suggested to be related to mucosal protection. We therefore investigated the importance of the removable mucous layer and epithelial bicarbonate transport in preservation of the gastric juxtamucosal pH (pHjm) during luminal acid. Anesthetized rats were prepared for intravital microscopy of the gastric mucosa, and pHjmwas measured with pH-sensitive microelectrodes. The mucus was either left intact (IM) or removed (MR) down to the firmly attached mucous layer, and HCl (pH 1) was applied luminally. Removal of the loosely adherent mucous layer did not influence the pHjmduring luminal acid (pentagastrin: IM/MR 7.03 ± 0.09/6.82 ± 0.19; pentagastrin + indomethacin: IM/MR 6.89 ± 0.20/6.95 ± 0.27; ranitidine: IM/MR 2.38 ± 0.64/2.97 ± 0.62), unless prostaglandin synthesis and acid secretion were inhibited (ranitidine + indomethacin: IM/MR 2.03 ± 0.37/1.66 ± 0.18). Neutral pHjmis maintained during endogenous acid secretion and luminal pH 1, unless DIDS was applied luminally, which resulted in a substantially decreased pHjm(1.37 ± 0.21). Neutral pHjmis maintained by a DIDS-sensitive bicarbonate transport over the surface epithelium. The loosely adherent mucous layer only contributes to maintaining pHjmduring luminal pH 1 if acid secretion and prostaglandin synthesis are inhibited.

Background The secreted proteins from goblet cells compose the intestinal mucus. The aims of this study were to determine how they exist in two intestinal mucus layers. Methodology/Principal Findings The intestinal mucosa was fixed with Carnoy solution and immunostained. Mucus from the loose layer, the firm layer was gently suctioned or scraped, respectively, lysed in SDS sample buffer with or without DTT, then subjected to the western blotting of rTFF3, rIgGFcγBP or rMuc2. The non-reduced or reduced soluble mucus samples in RIPA buffer were co-immunoprecipitated to investigate their possible interactions. Polyclonal antibodies for rTFF3, the rIgGFcγBP C-terminal domain and the rMuc2 C-terminal domain confirmed their localization in the mucus layer and in the mucus collected from the rat intestinal loose layer or firm layer in both western blot and immunoprecipitation experiments. A complex of rTFF3, which was approximately 250 kDa, and a monomer of 6 kDa were present in both layers of the intestinal mucus; rIgGFcγBP was present in the complex (250–280 kDa) under non-reducing conditions, but shifted to 164 kDa under reducing conditions in both of the layers. rMuc2 was found mainly in a complex of 214–270 kDa under non-reducing conditions, but it shifted to 140 kDa under reducing conditions. The co-immunoprecipitation experiments showed that binding occurs among rTFF3, rIgGFcγBP and rMuc2 in the RIPA buffer soluble intestinal mucus. Blocking the covalent interaction by 100 mM DTT in the RIPA buffer soluble intestinal mucus disassociated their binding. Conclusions/Significance Rat goblet cell-secreted TFF3, IgGFcγBP and Muc2, existing in the two intestinal mucus layers, are bound together by covalent interactions in the soluble fraction of intestinal mucus and form heteropolymers to be one of the biochemical mechanisms of composing the net-like structure of mucus.

The external mucus layers of the stony coral Porites astreoides and the soft corals Palythoa sp. and Heteroxenia fuscesens are inhabited by communities of marine heterotrophic bacteria. Population levels of bacteria in coral mucus may be regulated by the self-cleaning behavior of the host. Bacterial populations in coral mucus respond to stresses applied to the host coral by growing to higher population levels in the mucus, indicating that these are populations of viable organisms closely attuned to host metabolism. Members of these microbial populations utilize the mucus compounds and may play a role in processing coral mucus for reef detritus feeders. One such species, Vibrio alginolyticus, grows rapidly on Heteroxenia mucus, is attracted to dissolved mucus, and possesses a mechanism to maintain itself on the coral surface.

Mucus from the footsole of the freshwater snail Lymnaea stagnate behaves as a weak, negatively charged ion exchanger. Activities and concentrations of Na, K, Ca, and Cl were measured in mucus dialysed to equilibrium against artificial pond water or physiological saline. Observed activity coefficients (activity/concentration) in mucus were compared with those predicted by the Debye-Huckel theory to interpret the effects of electrostatic forces between the polyelectrolyte ions and small ions. The affinity of mucus for small ions decreased in the series, Ca2+, K+, Na+, Cl−. The extent to which mucus can concentrate cations was measured using three different methods: by titrating the fixed acidic groups with K or Ca and by equilibrium dialysis after which the electrical potential difference was either measured directly or was calculated from the Nernst potential for Na. Ion exchange titration indicated a much smaller exchange capacity than did the other two methods. Kinetics of cation uptake by the snail from dilute media were re-interpreted by considering the enhanced concentrations of cations in the mucus layer. It was shown that the presence of mucus in the unstirred layer adjacent to a transporting epithelium can result in an underestimate of the Michaelis constant (Km) determined from influx measurements.

The colonic epithelium facilitates host–microorganism interactions to control mucosal immunity, coordinate nutrient recycling and form a mucus barrier. Breakdown of the epithelial barrier underpins inflammatory bowel disease (IBD). However, the specific contributions of each epithelial-cell subtype to this process are unknown. Here we profile single colonic epithelial cells from patients with IBD and unaffected controls. We identify previously unknown cellular subtypes, including gradients of progenitor cells, colonocytes and goblet cells within intestinal crypts. At the top of the crypts, we find a previously unknown absorptive cell, expressing the proton channel OTOP2 and the satiety peptide uroguanylin, that senses pH and is dysregulated in inflammation and cancer. In IBD, we observe a positional remodelling of goblet cells that coincides with downregulation of WFDC2—an antiprotease molecule that we find to be expressed by goblet cells and that inhibits bacterial growth. In vivo, WFDC2 preserves the integrity of tight junctions between epithelial cells and prevents invasion by commensal bacteria and mucosal inflammation. We delineate markers and transcriptional states, identify a colonic epithelial cell and uncover fundamental determinants of barrier breakdown in IBD. Profiling of single epithelial cells in healthy and inflamed colons identifies specialized cellular subpopulations, including a type of goblet cell that secretes the antibacterial protein WFDC2, which preserves the integrity of the epithelial barrier layer.

The control of transport through mucus layers is a ubiquitous phenomenon in physiological systems. Mucus is often tasked with the mediation of passive, diffusive transport of small ionic species. However, questions remain regarding how mucin gel characteristics (charge density of the polymeric network, binding affinity of ions with mucus) govern the rate at which ions diffuse through mucus layers. Experimental studies measuring hydrogen diffusion through gastric mucus have provided conflicting results, and it is not clear if the rate of ionic diffusion through mucus layers is appreciably different than in aqueous environments (depending on experimental preparation). Here, we present a mathematical analysis of electrodiffussion of two ionic species (hydrogen and chloride) through a mucus layer. In addition to accounting for the chemical binding of hydrogen to the mucus network, we enforce a zero net current condition (as mucus layers in physiological systems are not generally electrogenic) and calculate the Donnan potential that occurs at the edge of the mucus layer. The model predicts the steady-state fluxes of ionic species and the induced potential across the layer. We characterize the dependence of these quantities on the chemical properties of the mucus gel, the composition of the bath solution, and the molecular mobility of the dissolved anion, and we show that the model predictions are consistent with a large portion of the experimental literature. Our analysis predicts that mucus layers generically slow the diffusive transport of hydrogen, but that chemical binding with the network attenuates this effect.

Induced goblet cells are useful for studying the interaction of intestinal mucus layer and the gut microbiota., Intestinal goblet cells secrete mucus layers protecting the intestinal epithelia against injuries. It is challenging to study the interaction of goblet cells, mucus layers, and gut microbiota because of difficulty in producing goblet cells and mucus models. We generate intestinal goblet cells from human epidermal stem cells with two small molecular inhibitors Repsox and CHIR99021 in the presence of basic fibroblast growth factor and bone morphogenetic protein 4 at high efficiency (~95%) of conversion for a short time (6 to 8 days). Induced goblet cells are functional to secrete mucus, deliver fluorescent antigen, and form mucus layers modeling the mucus-microbe interaction in vitro. Transplantation of induced goblet cells and oral administration of chemical induction media promote the repair of the intestinal epithelia in a colitis mouse model. Thus, induced goblet cells can be used for investigating mucus-microbe interaction, and chemical cocktails may act as drugs for repairing the intestinal epithelia.

Mammalian gut microbiota are integral to host health. However, how this association began remains unclear. We show that in basal chordates the gut space is radially compartmentalized into a luminal part where food microbes pass and an almost axenic peripheral part, defined by membranous delamination of the gut epithelium. While this membrane, framed with chitin nanofibers, structurally resembles invertebrate peritrophic membranes, proteome supports its affinity to mammalian mucus layers, where gut microbiota colonize. In ray-finned fish, intestines harbor indigenous microbes, but chitinous membranes segregate these luminal microbes from the surrounding mucus layer. These data suggest that chitin-based barrier immunity is an ancient system, the loss of which, at least in mammals, provided mucus layers as a novel niche for microbial colonization. These findings provide a missing link for intestinal immune systems in animals, revealing disparate mucosal environment in model organisms and highlighting the loss of a proven system as innovation., The coevolution of the animal gut mucosa and the gut microbiota is poorly understood. Here, Nakashima et al. identify intestinal chitinous membranes in basal chordates and ray-finned fish, and propose that the loss of this chitin barrier allowed mucus layers to become colonized by microbes in mammals.

The resistance of Helicobacter pylori (H. pylori) to conventional antibiotic treatments becomes prevalent recently. The biofilm formation was found to be highly correlated with the antibiotic resistance of H. pylori in the last decades. Moreover, H. pylori colonizes on the digestive tract epithelium located under the mucus layers, which further reduces therapeutic efficacy as mucus layers trap and remove exogenous substances including drugs. Herein, we reported a novel lipid polymer nanoparticles (LPNs) to overcome both biofilm and mucus layers obstruction. LPNs employed chitosan nanoparticle (CS NPs) as the core, mixed lipid layer containing rhamnolipids (RHL) as the shell and the surface of LPNs was further modified with DSPE-PEG2000 to improve hydrophilicity. Clarithromycin (CLR), a first-line drug for H. pylori infection, was encapsulated in LPNs. LPNs, especially the formulation utilizing 100% of RHL as the lipid shell, exhibited excellent eradicating ability to H. pylori biofilm, which was mainly reflected in the significant reduction of biofilm biomass and viability, destruction of biofilm architecture and elimination of extracellular polymeric substances (EPS). The anti-biofilm activities of LPNs are related to: 1) the disrupting effect of RHL on biofilm matrix; 2) antibacterial effects of CLR and CS NPs on biofilm bacteria and 3) inhibitory effects of CS NPs and RHL on bacteria adhesion and biofilm formation. Furthermore, PEGylated LPNs could rapidly penetrate through mucus without interacting with mucins and effectively eradicate H. pylori biofilm under mucus layer. In conclusion, a novel approach of drug-containing LPNs that could penetrate through mucus layers and effectively eradicate H. pylori biofilm provides new ways to treat persistent H. pylori infections.

Colonic epithelial cells are covered by thick inner and outer mucus layers. The inner mucus layer is free of commensal microbiota, which contributes to the maintenance of gut homeostasis. In the small intestine, molecules critical for prevention of bacterial invasion into epithelia such as Paneth-cell-derived anti-microbial peptides and regenerating islet-derived 3 (RegIII) family proteins have been identified. Although there are mucus layers providing physical barriers against the large number of microbiota present in the large intestine, the mechanisms that separate bacteria and colonic epithelia are not fully elucidated. Here we show that Ly6/PLAUR domain containing 8 (Lypd8) protein prevents flagellated microbiota invading the colonic epithelia in mice. Lypd8, selectively expressed in epithelial cells at the uppermost layer of the large intestinal gland, was secreted into the lumen and bound flagellated bacteria including Proteus mirabilis. In the absence of Lypd8, bacteria were present in the inner mucus layer and many flagellated bacteria invaded epithelia. Lypd8(-/-) mice were highly sensitive to intestinal inflammation induced by dextran sulfate sodium (DSS). Antibiotic elimination of Gram-negative flagellated bacteria restored the bacterial-free state of the inner mucus layer and ameliorated DSS-induced intestinal inflammation in Lypd8(-/-) mice. Lypd8 bound to flagella and suppressed motility of flagellated bacteria. Thus, Lypd8 mediates segregation of intestinal bacteria and epithelial cells in the colon to preserve intestinal homeostasis.

Background:: Helicobacter pylori is recognized as the most common cause of chronіc gastrіtis, and also an important pathogenic factor in peptic ulcer disease, after colonization and persistant in the mucus layers of the human stomach causes a raised level of pro-inflammatory cytokine such as IL-6. Materials and Methods: The study was conducted in Kirkuk city from 15th of December 2016 to 15th of June 2017 in Azady teaching hospital in Kirkuk governorate, anti H. pylori IgG level and IL-6 level were estimated by using anti H. pylori IgG ELISA kit and IL-6 ELISA kit. Results: A total of 90 patients the interleukin-6 level and anti H. pylori IgG level in a group of untreated patients against H. pylori infection (225.592 ng/L, 1.338 pg/ml respectively) and in a group of patients who had previously treated against H. pylori infection (102.60 ng/L, 0.350 pg/ml respectively) were significantly higher as compared to those of the control group (55.91 ng/L, 0.250 pg/ml respectively). Conclusion: There were a positive correlation between specific anti H. pylori IgG level and IL-6 level in H. pylori associated with gastric ulcer diseases.

Background & Aims Inflammation, injury, and infection up-regulate expression of the tryptophan metabolizing enzyme indoleamine 2,3-dioxygenase 1 (IDO1) in the intestinal epithelium. We studied the effects of cell-specific IDO1 expression in the epithelium at baseline and during intestinal inflammation in mice. Methods We generated transgenic mice that overexpress fluorescence-tagged IDO1 in the intestinal epithelium under control of the villin promoter (IDO1-TG). We generated intestinal epithelial spheroids from mice with full-length Ido1 (controls), disruption of Ido1 (knockout mice), and IDO1-TG and analyzed them for stem cell and differentiation markers by real-time polymerase chain reaction, immunoblotting, and immunofluorescence. Some mice were gavaged with enteropathogenic Escherichia coli (E2348/69) to induce infectious ileitis, and ileum contents were quantified by polymerase chain reaction. Separate sets of mice were given dextran sodium sulfate or 2,4,6-trinitrobenzenesulfonic acid to induce colitis; intestinal tissues were analyzed by histology. We utilized published data sets GSE75214 and GDS2642 of RNA expression data from ilea of healthy individuals undergoing screening colonoscopies (controls) and patients with Crohn’s disease. Results Histologic analysis of small intestine tissues from IDO1-TG mice revealed increases in secretory cells. Enteroids derived from IDO1-TG intestine had increased markers of stem, goblet, Paneth, enteroendocrine, and tuft cells, compared with control enteroids, with a concomitant decrease in markers of absorptive cells. IDO1 interacted non-enzymatically with the aryl hydrocarbon receptor to inhibit activation of NOTCH1. Intestinal mucus layers from IDO1-TG mice were 2-fold thicker than mucus layers from control mice, with increased proportions of Akkermansia muciniphila and Mucispirillum schaedleri. Compared to controls, IDO1-TG mice demonstrated an 85% reduction in ileal bacteria (P = .03) when challenged with enteropathogenic E coli, and were protected from immune infiltration, crypt dropout, and ulcers following administration of dextran sodium sulfate or 2,4,6-trinitrobenzenesulfonic acid. In ilea of Crohn’s disease patients, increased expression of IDO1 correlated with increased levels of MUC2, LYZ1, and aryl hydrocarbon receptor, but reduced levels of SLC2A5. Conclusions In mice, expression of IDO1 in the intestinal epithelial promotes secretory cell differentiation and mucus production; levels of IDO1 are positively correlated with secretory cell markers in ilea of healthy individuals and Crohn’s disease patients. We propose that IDO1 contributes to intestinal homeostasis.

Whether mucus layers lining entrance points into the body, including the lung airways, provide protection against the penetration of engineered nanoparticles remains poorly understood. We measured the diffusion coefficients of hundreds of individual nanoparticles of three different metal oxides (nMeOs) and two types of single-walled carbon nanotubes (SWCNTs) in undiluted human mucus. We found that the vast majority of these nanoparticles are efficiently trapped in human mucus and, further, that the mechanism of trapping is adhesive interactions as opposed to steric obstruction. However, a small fraction of zinc oxide (ZnO) nanoparticles moved at rates fast enough to penetrate airway mucus layers. We conclude that human mucus layers probably provide considerable protection for mucosal tissues from the penetration of most nMeOs and SWCNTs, and suggest that further investigation of the potential health risks of exposure to ZnO nanoparticles is warranted.

The MUC2 mucin polymer is the main building unit of the intestinal mucus layers separating intestinal microbiota from the host epithelium. The MUC2 mucin is a large glycoprotein with a C-terminal domain similar to the MUC5AC and MUC5B mucins and the von Willebrand factor (VWF). A structural model of the C-terminal part of MUC2, MUC2-C, was generated by combining Cryo-electron microscopy, AlphaFold prediction, information of its glycosylation, and small angle X-ray scattering information. The globular VWD4 assembly in the N-terminal of MUC2-C is followed by 3.5 linear VWC domains that form an extended flexible structure before the C-terminal cystine-knot. All gel-forming mucins and VWF form tail-tail disulfide-bonded dimers in their C-terminal cystine-knot domain, but interestingly the MUC2 mucin has an extra stabilizing disulfide bond on the N-terminal side of the VWD4 domain, likely essential for a stable intestinal mucus barrier.

Mammalian gut microbiota are integral to host health. However, how this association began remains unclear. We show that in basal chordates the gut space is radially compartmentalized into a luminal part where food microbes pass and an almost axenic peripheral part, defined by membranous delamination of the gut epithelium. While this membrane, framed with chitin nanofibers, structurally resembles invertebrate peritrophic membranes, proteome supports its affinity to mammalian mucus layers, where gut microbiota colonize. In ray-finned fish, intestines harbor indigenous microbes, but chitinous membranes segregate these luminal microbes from the surrounding mucus layer. These data suggest that chitin-based barrier immunity is an ancient system, the loss of which, at least in mammals, provided mucus layers as a novel niche for microbial colonization. These findings provide a missing link for intestinal immune systems in animals, revealing disparate mucosal environment in model organisms and highlighting the loss of a proven system as innovation.

Mucus layers (McLs) are on the front line of the human defense system that protect us from foreign abiotic/biotic particles (e.g., airborne virus SARS-CoV-2) and lubricates our organs. Recently, the impact of McLs on human health (e.g., nutrient absorption and drug delivery) and diseases (e.g., infections and cancers) has been studied extensively, yet their mechanisms are still not fully understood due to their high variety among organs and individuals. We characterize these variances as the heterogeneity of McLs, which lies in the thickness, composition, and physiology, making the systematic research on the roles of McLs in human health and diseases very challenging. To advance mucosal organoids and develop effective drug delivery systems, a comprehensive understanding of McLs' heterogeneity and how it impacts mucus physiology is urgently needed. When the role of airway mucus in the penetration and transmission of coronavirus (CoV) is considered, this understanding may also enable a better explanation and prediction of the CoV's behavior. Hence, in this Review, we summarize the variances of McLs among organs, health conditions, and experimental settings as well as recent advances in experimental measurements, data analysis, and model development for simulations.

The aim of this study was to investigate whether a mucus layer covers the surface of various animal feces and, if so, to show the structure and mucin composition of this layer. The freshly excreted feces of mice, rats, guinea pigs, rabbits and humans were fixed with Carnoy solution. Cross-sections approximately 5 microm thick were stained with alcian blue (AB) or the periodic acid Schiff (PAS) reaction. We measured the thickness of the mucus layer on the fecal surface of these sections. The fecal surface was covered with a continuous mucus layer in all specimens. The mucus layers of mice, rats and humans consisted of the alternate stratification of AB-positive and PAS-positive mucin layers. In contrast, the mucus layer consisted of an inner PAS-positive neutral mucin layer and an outer AB-positive acidic mucin layer in guinea pigs and rabbits. The average thicknesses of the mucus layers upon the fecal surfaces for mice, rats, guinea pigs, rabbits and humans were 19 +/- 12, 22 +/- 14, 15 +/- 4.6, 19 +/- 14 and 17 +/- 11 microm, respectively. These results demonstrated that the fecal surfaces are covered with continuous mucus layers in rodents, rabbits and humans, with substructures varying among species.

Under normal conditions, the vocal fold mucus layer is too thin to permit observation using videolaryngostroboscopy (VLS) during phonation. However, vocal nodules (VNs) typically cause congealed and sticky mucus to appear on the vocal fold. Reports in the literature regarding this phenomenon are limited. The aim of this study was to review VLS recordings of VN patients, analyzing changes that occurred in the mucus layer that covers the vocal fold during vibration following VN surgery.Using VLS, we studied the occurrence of, and changes in, vocal fold mucus layers in 160 VN patients before and after surgery.Eighty-eight patients (55%) were found to have a mucus layer during preoperative examinations. Of these mucus layers, 21 (13%) were located on the anterior commissure or anterior third of the vocal fold (A), 58 (36.3%) on the junction of the anterior and middle thirds (M), 1 (0.6%) on the posterior third (P), 5 (3.1%) on both A and M and 3 (1.9%) on both M and P. Fifty-six (35%) cases were found to have a mucus layer during postoperative examinations. Of these, 44 (27.5%) were located on A, 8 (5%) on M, 1 (0.6%) on P and 3 (1.9%) on both A and M. These results indicate that changes in the mechanical force on the vocal fold, alteration of the laryngeal secretory gland and improper aerodynamic airflow result in increased mucus viscosity and aggregation in VN patients and that the combination of these factors further increases the severity of dysphonia.Surgery to remove vocal nodes may be an effective method to eliminate both vocal bumps and aggregated mucus. Based on the present results, it is recommended that future research should compare surgery to remove VN against other mucus layer reduction methods in order to determine which is the most effective.

Significance: Hydrogen sulfide (H2S), an important regulator of physiology and health, helps resolve inflammation and promotes tissue repair in the gastrointestinal tract. Recent Advances: Gut microbiota live as a multispecies biofilm in close interaction with the upper mucus layer lining the epithelium. The relative abundance, spatial organization, and function of these microorganisms affect a broad range of health outcomes. This article provides a state-of-the-art review of our understanding of the cross talk between H2S, the gut microbiota, and health. H2S can have toxic or therapeutic effects, depending on its concentration and source. When produced at excessive concentrations by local microbiota, H2S may cause mucus disruption and inflammation and contribute to development of cancer. In contrast, low levels of endogenous or exogenous H2S directly stabilize mucus layers, prevent fragmentation and adherence of the microbiota biofilm to the epithelium, inhibit the release of invasive pathobionts, and help resolve inflammation and tissue injury. Although scarce, research findings suggest that dietary H2S obtained from plants or ingestion of the H2S precursor, L-cysteine, may also modulate the abundance and function of microbiota. Critical Issues: A critical issue is the lack of understanding of the metagenomic, transcriptomic, and proteomic alterations that characterize the interactions between H2S and gut microbiota to shape health outcomes. Future Directions: The ambivalent roles of H2S in the gut offer a fertile ground for research on such critical issues. The findings will improve our understanding of how H2S modulates the microbiota to affect body function and will help identify novel therapeutic strategies.

Colonic mucus layers protect intestinal tissues against intestinal bacteria. We investigated the effects of dietary fiber and its metabolites on mucus production in the colonic mucosa. Mice were fed a partially hydrolyzed guar gum (PHGG)-containing diet and a fiber-free diet (FFD). The colon mucus layer, fecal short-chain fatty acid (SCFA) levels, and gut microbiota were evaluated. Mucin 2 (MUC2) expression was assessed in SCFA-treated LS174T cells. The role of AKT in MUC2 production was investigated. The mucus layer in the colonic epithelium was significantly increased in the PHGG group compared with that in the FFD group. In the PHGG group, an increase in Bacteroidetes in the stool was observed, and fecal acetate, butyrate, propionate, and succinate levels were significantly increased. However, MUC2 production was significantly increased only in succinate-stimulated LS174T cells. The succinate-induced MUC2 production was associated with AKT phosphorylation. Succinate mediated the PHGG-induced increase in the colon mucus layer.

Background: Helicobacter pylori recognized as the most common cause of chronic gastritis and an important pathogenic factor in peptic ulcer, after colonization and stomach mucus layers caused increase in the levels of anti-inflammatory cytokine such as IL-13. The aims of this study is compare of IL-13 levels among treated patients with H. Pylori gastric infection, patients with H. Pylori gastric infection without treatment and healthy person Materials and Methods: The study was conducted in Kirkuk city from the September 2020 to March 2020, ELISA test used for interleukin -13 with peptic ulcer diseases (PUD). Results: Elevated levels of IL-13 reported in patients with acute and chronic H. pylori gastric infection compared to control group (44.74+8.18). A moderate significant in IL-13 level determined in infected patients without treatment, previously treated patients (140.12+45.36 vs 105+22.71). While, there was a high significant difference when these two groups and compared with the control group (44.74+8.18)). Conclusion: IL-13 levels is highest in patients with H. pylori gastric infection in compare healthy person.

The mucus layer coating the gastrointestinal tract is the front line of innate host defense, largely because of the secretory products of intestinal goblet cells. Goblet cells synthesize secretory mucin glycoproteins (MUC2) and bioactive molecules such as epithelial membrane-bound mucins (MUC1, MUC3, MUC17), trefoil factor peptides (TFF), resistin-like molecule beta (RELMbeta), and Fc-gamma binding protein (Fcgbp). The MUC2 mucin protein forms trimers by disulfide bonding in cysteine-rich amino terminal von Willebrand factor (vWF) domains, coupled with crosslinking provided by TFF and Fcgbp proteins with MUC2 vWF domains, resulting in a highly viscous extracellular layer. Colonization by commensal intestinal microbiota is limited to an outer "loose" mucus layer, and interacts with the diverse oligosaccharides of mucin glycoproteins, whereas an "inner" adherent mucus layer is largely devoid of bacteria. Defective mucus layers resulting from lack of MUC2 mucin, mutated Muc2 mucin vWF domains, or from deletion of core mucin glycosyltransferase enzymes in mice result in increased bacterial adhesion to the surface epithelium, increased intestinal permeability, and enhanced susceptibility to colitis caused by dextran sodium sulfate. Changes in mucin gene expression and mucin glycan structures occur in cancers of the intestine, contributing to diverse biologic properties involved in the development and progression of cancer. Further research is needed on identification and functional significance of various components of mucus layers and the complex interactions among mucus layers, microbiota, epithelial cells, and the underlying innate and adaptive immunity. Further elucidation of the regulatory mechanisms involved in mucin changes in cancer and inflammation may lead to the development of novel therapeutic approaches.

Akkermansia muciniphila is a probiotic inhabiting host intestinal mucus layers and displays evident easing or therapeutic effects on host enteritis and metabolic disorders such as obesity and diabetes. The outer membrane protein Amuc_1100 of A. muciniphila is likely to play a crucial role during the interaction with the host. 5-HT is a neurotransmitter and a key signal molecule regulating the gastrointestinal tract functions and other organs, which is involved in diverse physiological and pathological processes. This study demonstrated that Amuc_1100 could promote the expression of the 5-HT synthesis rate-limiting enzyme Tph1 in RIN-14B cells and reduce the expression of the serotonin reuptake transporter (SERT) in Caco-2 cells through direct interaction with TLR2, thereby improving 5-HT biosynthesis and extracellular availability. Using antibiotic-treated mice as animal models, we found that after gavage with A. muciniphila or Amuc_1100, Tph1 expression increased and SERT expression decreased in colon tissues. The 5-HT concentrations in colon tissues and blood were markedly elevated simultaneously. We also found that A. muciniphila or Amuc_1100 improved the gastrointestinal motility function and restored gut microbiota abundance and species diversity in antibiotic-treated mice. These results suggest that A. muciniphila can regulate the host intestinal 5-HT system via its outer membrane protein Amuc_1100 and TLR2. This mechanism represented an important approach through which A. muciniphila interacts with the host and further influences 5-HT-related physiological functions. These results advance the understanding of interplay mechanisms between the gut microbiota and the host, which could be the basis for new intervention strategies for related diseases.

Diffusive transport of small ionic species through mucus layers is a ubiquitous phenomenon in physiology. However, some debate remains regarding how the various characteristics of mucus (charge of the polymers themselves, binding affinity of ions with mucus) impact the rate at which small ions may diffuse through a hydrated mucus gel. Indeed it is not even clear if small ionic species diffuse through mucus gel at an appreciably different rate than they do in aqueous solution. Here, we present a mathematical description of the transport of two ionic species (hydrogen and chloride) through a mucus layer based on the Nernst-Planck equations of electrodiffusion. The model explicitly accounts for the binding affinity of hydrogen to the mucus material, as well as the Donnan potential that occurs at the interface between regions with and without mucus. Steady state fluxes of ionic species are quantified, as are their dependencies on the chemical properties of the mucus gel and the composition of the bath solution. We outline a mechanism for generating enhanced diffusive flux of hydrogen across the gel region, and hypothesize how this mechanism may be relevant to the apparently contradictory experimental data in the literature.

Secretory immunoglobulin A (sIgA), a dimeric antibody found in high quantities in the gastrointestinal mucosa, is broadly associated with mucosal immune protection. A distinguishing feature of sIgA is its ability to crosslink pathogens, thereby creating pathogen/sIgA aggregates that are too large to traverse the dense matrix of mucin fibers in mucus layers overlying epithelial cells and consequently reducing infectivity. Here, we use modeling to investigate this mechanism of “immune exclusion” based on sIgA-mediated agglutination, in particular the potential use of sIgA to agglutinate HIV in cervicovaginal mucus (CVM) and prevent HIV transmission. Utilizing reported data on HIV diffusion in CVM and semen, we simulate HIV collision kinetics in physiologically-thick mucus layers–a necessary first step for sIgA-induced aggregation. We find that even at the median HIV load in semen of acutely infected individuals possessing high viral titers, over 99% of HIV virions will penetrate CVM and reach the vaginal epithelium without colliding with another virion. These findings imply that agglutination is unlikely to be the dominant mechanism of sIgA-mediated protection against HIV or other sexually transmitted pathogens. Rather, we surmise that agglutination is most effective against pathogens either present at exceedingly high concentrations or that possess motility mechanisms other than Brownian diffusion that significantly enhance encounter rates.

The biological fate of polymeric micelles (PMs) following oral administration was investigated in this study to better understand the contribution of transport of integral PMs to oral absorption. To track integral PMs, near-infrared fluorophores with aggregation-caused quenching properties were utilized to label PMs comprised of methoxy poly(ethylene glycol)-poly(D,L-lactic acid) (mPEG-PDLLA) copolymers and methoxy poly(ethylene glycol)-distearoyl phosphoethanolamine (DSPE-PEG). The particle size of PMs prepared from mPEG2.5k-PDLLA1.25k, mPEG2.5k-PDLLA2.5k, mPEG5k-PDLLA3k, mPEG5k-PDLLA5k and DSPE-PEG2k was 24.5, 29.5, 34.0, 41.4 and 15.6 nm, respectively. After oral administration by gavage to rats, PMs were retained in the gastrointestinal tract for at least 4 h, and the copolymer block chain lengths did not have significant influence. The emergence of fluorescence in the blood and liver served as direct evidence to support oral absorption of integral PMs. Approximately 1–2% of intact particles were absorbed via the lymphatic pathway, but the total amount of PMs that reach the systemic circulation await further elucidation. Confocal laser scanning microscopy added more evidence to support the penetration of integral PMs into the basolateral tissues of microvilli. Cellular uptake efficiency was about 4–7% in Caco-2 cell lines for all PM groups, but was reduced to 1–3% in Caco-2/HT29-MTX co-culture models due to the hindrance by the mucus layers. Approximately 6–12% of integral PMs were transported across Caco-2/HT29-MTX/Raji monolayers, whereas only approximately one-tenth of that amount was transported across Caco-2 and Caco-2/HT29-MTX monolayers. Differences, but not statistically significant, were observed between PM groups in lymphatic uptake, biodistribution, cellular uptake and trans-monolayer transport, possibly owing to difference in block chain lengths as well as particle size. In conclusion, evidence obtained in this study supports penetration of integral PMs across the enteric epithelia, but the total amount may be limited.

The intestinal mucosal barrier, which is composed of epithelial cells and mucus layers secreted by goblet cells and contains commensal bacteria, constitutes the first line of defense against pathogenic gut microbiota. However, homeostasis between the microbiota and mucus layer is easily disrupted by certain factors, resulting in alteration of the gut microbiota and entry of pathogens to the intestinal mucosal barrier. In this review, we describe the structures and functions of the mucus layer, expound several crucial influencing factors, including diet styles, medications and host genetics, and discuss how pathogenic microorganisms interact with the mucus layer and commensal microbiota, with the understanding that unraveling their complex interactions under homeostatic and dysbiosis conditions in the colon would help reveal some underlying pathogenic mechanisms and thus develop new strategies to prevent pathogenic microbiological colonization.

Background & Aims Infections resulting from intestinal yeast and bacteria affect a large number of patients with deficits in absorptive or secretory epithelial transport mechanisms. The basolateral Na+-K+-2Cl- cotransporter (NKCC1) has been implicated in intestinal epithelial fluid secretion. Two patients with deleterious heterozygous (NKCC1-DFX, DFX for Asp-Phe-stop codon) or homozygous (Kilquist) mutations in SLC12A2 (NKCC1) suffered from gastrointestinal deficits. Because of chronic infections, the colon and the small intestine of the NKCC1-DFX patient were resected surgically. Methods To investigate how NKCC1 affects the integrity and function of the gut epithelia, we used a mouse model recapitulating the NKCC1-DFX patient mutation. Electron microscopy and immunostaining were used to analyze the integrity of the colonic mucus layers and immune cell infiltration. Fluorescence in situ hybridization was performed on the distal colon sections to measure bacteria translocation to the mucosa and submucosa. Citrobacter rodentium was used to measure mouse ability to clear enteric infection. A multiplex cytokine assay was used to analyze mouse inflammatory response to infection. Results We show that NKCC1-DFX expression causes defective goblet cell mucus granule exocytosis, leading to secretion of intact granules into the lumen of the large intestine. In addition, NKCC1-DFX colon submucosal glands secrete mucus that remained attached to the epithelium. Importantly, expression of the mutant NKCC1 or complete loss of NKCC1 function leads to aggravated inflammatory response to C rodentium infection. Compared with wild-type, NKCC1-DFX mice showed decreased expression of claudin-2, a tight junction protein involved in paracellular Na+ and water transport and enteric infection clearance. Conclusions Our data indicate that NKCC1-DFX impairs gut barrier function by affecting mucus secretion and immune properties., Graphical abstract

Oral delivery of therapeutics is the preferred route of administration due to ease of administration which is associated with greater patient medication adherence. One major barrier to oral delivery and intestinal absorption is rapid clearance of the drug and the drug delivery system from the gastrointestinal (GI) tract. To address this issue, researchers have investigated using GI mucus to help maximize the pharmacokinetics of the therapeutic; while mucus can act as a barrier to effective oral delivery, it can also be used as an anchoring mechanism to improve intestinal residence. Nano-drug delivery systems that use materials which can interact with the mucus layers in the GI tract can enable longer residence time, improving the efficacy of oral drug delivery. This review examines the properties and function of mucus in the GI tract, as well as diseases that alter mucus. Three broad classes of mucus-interacting systems are discussed: mucoadhesive, mucus-penetrating, and mucolytic drug delivery systems. For each class of system, the basis for mucus interaction is presented, and examples of materials that inform the development of these systems are discussed and reviewed. Finally, a list of FDA-approved mucoadhesive, mucus-penetrating, and mucolytic drug delivery systems is reviewed. In summary, this review highlights the progress made in developing mucus-interacting systems, both at a research-scale and commercial-scale level, and describes the theoretical basis for each type of system.

Mucin gene is expressed at the mucous membrane of the inner layer of the internal organs. Intestinal mucin 2 (MUC2), a major gel-forming mucin, represents a primary barrier component of mucus layers. This is the first report of role of mucin gene in growth traits in animals. In the current study, we had randomly studied Bengal ducks (Anas platyrynchos) reared from day old to 10 weeks of age under organized farm and studied the growth parameters as well as body weight and average daily body weight gain. We had characterized mucin gene for Bengal duck and observed glycosylation and EGF1 (EGF like domain signature) as important domain for growth traits in duck. We observed better expression profile for mucin gene in high growing ducks in comparison to that of lower growing ducks with real time PCR. Hence mucin gene may be employed as a marker for growth traits.