Mucus-secreting Goblet Cells Research Articles

A Mouse Model of Ovalbumin-Induced Airway Allergy Exhibits Altered Localization of SARS-CoV-2-Susceptible Cells in the Lungs, Which Reflects Omicron BA.5 Infection Dynamics, Viral Mutations, and Immunopathology.

Asthma, an allergic disease of the airways, is a risk factor for severity of common respiratory viral infections; however, the relationship between asthma and severity in COVID-19 remains unclear. Here, we examined the effects of SARS-CoV-2 (Omicron BA.5 strain) infection in a mouse model of airway allergy. First, stimulation of allergic mice with OVA resulted in the appearance of ACE2-negative mucus-secreting goblet cells in the bronchiolar region, and an increase in the number of ACE2-expressing cells in the alveoli. As a result, ACE2-expressing cells, which are susceptible to SARS-CoV-2, were limited to the distal portion of the bronchioles while they increased in the alveolar area. After viral infection, the peak infectious viral load in the OVA group was 100-fold lower than that in the phosphate buffered saline (PBS) group; however, clearance of viral RNA from the upper/lower airways was delayed. There were notable differences in acquisition of nsp5 and nsp6 mutations by the Omicron BA.5 strain recovered from BALF samples obtained from the OVA and PBS groups. Immune responses associated with viral clearance were essentially the same, but expression of granulocyte-associated chemokines was higher, M2 macrophage responses were predominant, and the higher spike-specific IgG1/IgG2a ratio in the OVA group post-infection. Infection localized in the alveolar region earlier in the OVA group, resulting in more severe alveolar damage than in the PBS group. These data suggest a Th2-shifted immune background and altered localization of SARS-CoV-2 susceptible cells in mice with OVA-induced airway allergy, which reflect Omicron BA.5 infection dynamics, viral mutations, and immunopathology.

Neonatal microbiota colonization drives maturation of primary and secondary goblet cell mediated protection in the pre-weaning colon.

In the distal colon, mucus secreting goblet cells primarily confer protection from luminal microorganisms via generation of a sterile inner mucus layer barrier structure. Bacteria-sensing sentinel goblet cells provide a secondary defensive mechanism that orchestrates mucus secretion in response to microbes that breach the mucus barrier. Previous reports have identified mucus barrier deficiencies in adult germ-free mice, thus implicating a fundamental role for the microbiota in programming mucus barrier generation. In this study, we have investigated the natural neonatal development of the mucus barrier and sentinel goblet cell-dependent secretory responses upon postnatal colonization. Combined in vivo and ex vivo analyses of pre- and post-weaning colonic mucus barrier and sentinel goblet cell maturation demonstrated a sequential microbiota-dependent development of these primary and secondary goblet cell-intrinsic protective functions, with dynamic changes in mucus processing dependent on innate immune signalling via MyD88, and development of functional sentinel goblet cells dependent on the NADPH/Dual oxidase family member Duox2. Our findings therefore identify new mechanisms of microbiota-goblet cell regulatory interaction and highlight the critical importance of the pre-weaning period for the normal development of colonic barrier function.

Histological features of the gastrointestinal tract of elongate tigerfish, Hydrocynus forskahlii (Cuvier, 1819), from Lake Albert

BackgroundThe tigerfish (Hydrocynus forskahlii) is an important food fish in different regions of Africa. As such, interest in its performance and nutritional requirements as a potential candidate for aquaculture is increasing. Characterization of the morpho-histological features and functions of the gut provides valuable insights into the feeding physiology and digestive system of fish species.ResultAn investigation of the morpho-histological features of the gastrointestinal tract of H. forskahlii captured from Lake Albert was conducted over an eight-month period. The digestive tract of H. forskahlii is characterised by a short oesophagus, well-developed and distensible stomach and moderately long intestine. The oesophagus had a higher abundance of mucous-secreting goblet cells compared to other digestive tract tissues. The stomach had a bag-like shape for swallowing large prey and was distinguishable into the cardiac, fundic and pyloric regions. Both the cardiac and fundic regions of the stomach had numerous gastric glands, whereas the pyloric region had large compound folds consisting of pseudo-stratified columnar epithelium. Many goblet cells were also observed in the mid and posterior regions of the intestine. The relative length of the gut of the H. forskahlii was 1.27 ± 0.03 cm, and it had a total of 20–22 pyloric caeca. The liver was observed to be divided in lobules by a central vein contained within connective sheets. Irregular cords of hepatocytes were also visible throughout the parenchyma.ConclusionsThe histological features of the H. forskahlii gut observed in the present study are consistent with those of carnivorous and omnivorous fish.

24 Intestinal Function and Integrity in Health and Disease: Current Knowledge in Swine

Abstract Optimal lean tissue accretion is contingent on the ability of the pig to efficiently establish and maintain host defenses, and to digest and absorb nutrients and water. Epithelial cells in the mucosa of the gastrointestinal tract interconnect with each other by tight junctions, adherens junctions, and desmosomes, forming a complete intestinal barrier. Of the secretory cells in this epithelial barrier, mucus secreting goblet cells and hormone producing enteroendocrine cells can be found interspersed, while Paneth cells lie in the base of the crypts and secrete antimicrobial proteins/peptides into the mucus layer. In the small intestine, enterocytes lining the villi allow for efficient nutrient transport. Maintenance of the epithelial barrier and its function is dependent on intestinal stem cells residing at the crypt base. Two major signaling pathways controlling intestinal stem cell proliferation and differentiation are the β-catenin/Wnt and Notch signaling pathways. Thus, achieving and maintaining optimal nutrient absorptive capacity requires an intricate balance amongst β-catenin/Wnt and Notch signaling pathways, wherein dysregulation of either can result in inefficiency of nutrient absorption. Weaning transition, off-feed events, hypophagia, and enteric and respiratory diseases can all compromise the structural and secretory barrier of the gastrointestinal tract mucosa via changes in tight and adherens junction proteins and mucin production. As a result, these stressors can increase the incidence of gastric ulcers, villous atrophy, and colitis in pigs. Depending on the severity and duration of the mucosal damage, repair and restitution occurs due to increased stem cell proliferation, migration, and repolarization. Thus, efficiency of digestive and absorptive function depends on several factors, including the types of epithelial cells present and their functionality, the rate of cell renewal, and epithelial surface area. This paper will discuss what we know about the regulation and repair of the mucosal epithelium in growing pigs during health, stress, and disease.

Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation

In vitro culture and generation of highly specialized goblet cells is still a major challenge in conjunctival 3D in vitro equivalents. A model comprising all physiological factors, including mucus-secreting goblet cells has the potential to act as a new platform for studies on conjunctival diseases. We isolated primary conjunctival epithelial cells and fibroblasts from human biopsies. 3D models were generated from either epithelial layers or a combination of those with a connective tissue equivalent. Epithelial models were investigated for marker expression and barrier function. Full-thickness models were analyzed for goblet cell morphology and marker expression via immunofluorescence and quantitative real-time PCR. Simple epithelial models cultured at the air–liquid interface showed stratified multi-layer epithelia with pathologic keratinization and without goblet cell formation. The combination with a connective tissue equivalent to generate a full-thickness model led to the formation of a non-keratinized stratified multi-layer epithelium and induced goblet cell differentiation. In our model, a high resemblance to natural conjunctiva was achieved by the combination of conjunctival epithelial cells with fibroblasts embedded in a collagen-hydrogel as connective tissue equivalent. In the future, our conjunctival in vitro equivalent enables the investigation of goblet cell differentiation, conjunctival pathologies as well as drug testing.

Activating transcription factor 5 (ATF5) controls intestinal tuft and goblet cell expansion upon succinate-induced type 2 immune responses in mice.

Intestinal tuft cells, a chemosensory cell type in mucosal epithelia that secrete interleukin (IL)-25, play a pivotal role in type 2 immune responses triggered by parasitic infections. Tuft cell-derived IL-25 activates type 2 innate lymphoid cells (ILC2) to secrete IL-13, which, in turn, acts on intestinal stem or transient amplifying cells to expand tuft cells themselves and mucus-secreting goblet cells. However, the molecular mechanisms of tuft cell differentiation under type 2 immune responses remain unclear. The present study investigated the effects of the deletion of activating transcription factor 5 (ATF5) on the type 2 immune response triggered by succinate (a metabolite of parasites) in mice. ATF5 mRNAs were expressed in the small intestine, and the loss of the ATF5 gene did not affect the gross morphology of the tissue or the basal differentiation of epithelial cell subtypes. Succinate induced marked increases in tuft and goblet cell numbers in the ATF5-deficient ileum. Tuft cells in the ATF5-deficient ileum are assumed to be a subtype of intestinal tuft cells (Tuft-2 cells) marked by the transcription factor Spib. Exogenous IL-25 induced similar increases in tuft and goblet cell numbers in wild-type and ATF5-deficient ilea. IL-13 at a submaximal dose enhanced tuft cell differentiation more in ATF5-deficient than in wild-type intestinal organoids. These results indicate that the loss of ATF5 enhanced the tuft cell-ILC2 type 2 immune response circuit by promoting tuft cell differentiation in the small intestine, suggesting its novel regulatory role in immune responses against parasitic infections.

Effects of probiotics on growth, survival, and intestinal and liver morphometry of Gangetic mystus (Mystus cavasius)

The usage of probiotics proved advantageous in aquaculture due to its positive impact on fish growth, immune response and environment. This study was aimed to assess the effects of probiotics on growth, survival and histometry of intestine and liver in Gangetic mystus (Mystus cavasius) using two separate experiments for a period of 8 weeks (in aquaria) and 16 weeks (in earthen ponds). Three different probiotic treatments were incorporated i.e. commercial probiotic one; CP-1 (T1), commercial probiotic two; CP-2 (T2), Lab developed (Lab dev.) probiotic (T3) including a control. The results indicated that the probiotics usage especially Lab dev. probiotic (T3) significantly improved the growth parameters such as weight gain (g) and specific growth rate (SGR, %/day) as well as ensured better feed conversion efficiency. Zero mortality was observed in aquaria whereas probiotic application enhanced survivability in earthen ponds. Moreover, all probiotic treatment exhibited positive results for different histo-morphometric features of intestine and liver. Mucus secreting goblet cells and fattening of mucosal fold increased significantly with probiotic usage. The amount of regular shaped nucleus was maximum in T3 with least intra cellular distance between liver tissues in earthen ponds. The greatest value for hemoglobin with lowest glucose level was observed in T3 as well. Furthermore, probiotic ensured low concentration of ammonia during culture. Overall, it was anticipated that the application of probiotics in Gangetic mystus culture resulted positive effect on its growth, feed utilization, survivability, histo-morphometry, immunity and hematological parameters.

Morphology, Histology, and Histochemistry of the Digestive Tract of the Marbled Flounder Pseudopleuronectes yokohamae

This study investigated the morphological, histological, and histochemical characteristics of the digestive tract of the marbled flounder (Pseudopleuronectes yokohamae). The relative length of the gut of the marbled flounder digestive tract was 1.54 ± 0.10 (n = 20), and it had a simple stomach and 6-9 pyloric caeca. The mucosal folds of the marbled flounder digestive tract exhibited a general branched morphology. The thickness and mucosal fold length of the intestinal muscularis externa showed similar aspects in all areas. The thickness of the intestinal muscularis externa was the thickest in the posterior intestine portion, and the length of mucosal folds was the longest in the anterior intestine portion. It was indicated that food digested by gastric acid in the stomach moves to the anterior portion (including pyloric caeca) and mid portion of the intestine, ensuring effective stimulation of cholecystokinin (CCK)-producing cells. In addition, the distribution pattern of CCK-producing cells in the intestine was very similar to that of mucus-secreting goblet cells. The CCK-producing cells and goblet cells in the marbled flounder were well-adapted to promote optimal control of the digestive process. Based on the morphological and histochemical studies, it was concluded that the marbled flounder displays a digestive tract comparable to that of fish species with carnivorous habits.

The intestine: A highly dynamic microenvironment for IgA plasma cells.

To achieve longevity, IgA plasma cells require a sophisticated anatomical microenvironment that provides cytokines, cell-cell contacts, and nutrients as well as metabolites. The intestinal epithelium harbors cells with distinct functions and represents an important defense line. Anti-microbial peptide-producing paneth cells, mucus-secreting goblet cells and antigen-transporting microfold (M) cells cooperate to build a protective barrier against pathogens. In addition, intestinal epithelial cells are instrumental in the transcytosis of IgA to the gut lumen, and support plasma cell survival by producing the cytokines APRIL and BAFF. Moreover, nutrients are sensed through specialized receptors such as the aryl hydrocarbon receptor (AhR) by both, intestinal epithelial cells and immune cells. However, the intestinal epithelium is highly dynamic with a high cellular turn-over rate and exposure to changing microbiota and nutritional factors. In this review, we discuss the spatial interplay of the intestinal epithelium with plasma cells and its potential contribution to IgA plasma cell generation, homing, and longevity. Moreover, we describe the impact of nutritional AhR ligands on intestinal epithelial cell-IgA plasma cell interaction. Finally, we introduce spatial transcriptomics as a new technology to address open questions in intestinal IgA plasma cell biology.

Inhibition of neutrophil elastase prevents cigarette smoke exposure-induced formation of neutrophil extracellular traps and improves lung function in a mouse model of chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is an important public health challenge worldwide, and is usually caused by significant exposure to noxious agents, particularly cigarette smoke. Recent studies have revealed that excessive production of neutrophil extracellular traps (NETs) in the airways is associated with disease severity in COPD patients. NETs are extracellular neutrophil-derived structures composed of chromatin fibers decorated with histones and granule proteases including neutrophil elastase (NE). However, the effective prevention of NET formation in COPD remains elusive. Here, we demonstrated that treatment with GW311616A, a potent and selective inhibitor of NE, prevented cigarette smoke extract (CSE)-induced NET formation in human neutrophils by blocking NE nuclear translocation and subsequent chromatin decondensation. Inhibition of NE also abrogated CSE-induced ROS production and migration impairment of neutrophils. Administration of GW311616A in vivo substantially reduced pulmonary generation of NETs while attenuating the key pathological changes in COPD, including airway leukocyte infiltration, mucus-secreting goblet cell hyperplasia, and emphysema-like alveolar destruction in a mouse model of COPD induced by chronic cigarette smoke exposure. Mice treated with GW311616A also showed significant attenuation of neutrophil numbers and percentages and the levels of neutrophil chemotactic factors (LTB4, KC, and CXCL5) and proinflammatory cytokines (IL-1β, and TNF-α) in bronchoalveolar lavage fluid compared to mice treated with cigarette smoke exposure only. Furthermore, GW311616A treatment considerably improved lung function in the COPD mouse model, including preventing the decline of FEV100/FVC and delta PEF as well as inhibiting the increase in FRC, TLC, and FRC/TLC. Overall, our study suggests that NE plays a critical role in cigarette smoke-induced NET formation by neutrophils and that inhibition of NE is a promising strategy to suppress NET-mediated pathophysiological changes in COPD.

BMP gradient along the intestinal villus axis controls zonated enterocyte and goblet cell states.

Intestinal epithelial cells derive from stem cells at the crypt base and travel along the crypt-villus axis to die at the villus tip. The two dominant villus epithelial cell types, absorptive enterocytes and mucous-secreting goblet cells, are mature when they exit crypts. Murine enterocytes switch functional cell states during migration along the villus. Here, we ask whether this zonation is driven by the bone morphogenetic protein (BMP) gradient, which increases toward the villus. Using human intestinal organoids, we show that BMP signaling controls the expression of zonated genes in enterocytes. We find that goblet cells display similar zonation involving antimicrobial genes. Using an inducible Bmpr1a knockout mouse model, we confirm that BMP controls these zonated genes invivo. Our findings imply that local manipulation of BMP signal strength may be used to reset the enterocyte "rheostat" of carbohydrate versus lipid uptake and to control the antimicrobial response through goblet cells.

VEGF receptor 2 (KDR) protects airways from mucus metaplasia through a Sox9-dependent pathway.

Mucus-secreting goblet cells are the dominant cell type in pulmonary diseases, e.g., asthma and cystic fibrosis (CF), leading to pathologic mucus metaplasia and airway obstruction. Cytokines including IL-13 are the major players in the transdifferentiation of club cells into goblet cells. Unexpectedly, we have uncovered a previously undescribed pathway promoting mucous metaplasia that involves VEGFa and its receptor KDR. Single-cell RNA sequencing analysis coupled with genetic mouse modeling demonstrates that loss of epithelial VEGFa, KDR, or MEK/ERK kinase promotes excessive club-to-goblet transdifferentiation during development and regeneration. Sox9 is required for goblet cell differentiation following Kdr inhibition in both mouse and human club cells. Significantly, airway mucous metaplasia in asthmatic and CF patients is also associated with reduced KDR signaling and increased SOX9 expression. Together, these findings reveal an unexpected role for VEGFa/KDR signaling in the defense against mucous metaplasia, offering a potential therapeutic target for this common airway pathology.

Thyroid Hormones Regulate Goblet Cell Differentiation and Fgf19-Fgfr4 Signaling.

Hypothyroidism is a common pathological condition characterized by insufficient activity of the thyroid hormones (THs), thyroxine (T4), and 3,5,3'-triiodothyronine (T3), in the whole body or in specific tissues. Hypothyroidism is associated with inadequate development of the intestine as well as gastrointestinal diseases. We used a zebrafish model of hypothyroidism to identify and characterize TH-modulated genes and cellular pathways controlling intestine development. In the intestine of hypothyroid juveniles and adults, the number of mucus-secreting goblet cells was reduced, and this phenotype could be rescued by T3 treatment. Transcriptome profiling revealed dozens of differentially expressed genes in the intestine of hypothyroid adults compared to controls. Notably, the expression of genes encoding to Fgf19 and its receptor Fgfr4 was markedly increased in the intestine of hypothyroid adults, and treatment with T3 normalized it. Blocking fibroblast growth factor (FGF) signaling, using an inducible dominant-negative Fgfr transgenic line, rescued the number of goblet cells in hypothyroid adults. These results show that THs inhibit the Fgf19-Fgfr4 signaling pathway, which is associated with inhibition of goblet cell differentiation in hypothyroidism. Both the TH and Fgf19-Fgfr4 signaling pathways can be pharmaceutical targets for the treatment of TH-related gastrointestinal diseases.

Ciliated columnar epithelium pathomorphosis in children with upper and lower respiratory infections: ultrastructural and nanoscopic analysis

The cytomorphological profile of nasal epithelium in children with acute and chronic respiratory disorders was characterized. The redistribution of nasal ciliary epithelial cells in favor of the mucus-secreting (goblet) cells was observed (group with acute respiratory infection – ratio 2.3:1; group with chronic lung disease – 1:2.4) with normal values of these indicators 5:1 (control group). The mucosal metaplasia, against a background of local leukocyte infiltration, was detected among 28 patients (64.29 %). Using atomic force microscopy, the pathomorphosis of the cytoplasmic membrane ciliated epithelium was described, which characterized by the changes in roughness parameters (Ra, Rq, Rsk, Rmax, Rsk) and waviness (Wa) in group with chronic lung disease (Ra – 34.94 ± 7.8 nm, Rq – 41.26 ± 7.5, Rmax – 225.55 ± 44.43, Rsk – 1,2, Wa – 43.23 ± 12.4 nm) compared with control group (Ra – 7.22 ± 1.94 nm, Rq – 11.43 ± 1.83, Rmax – 111.83 ± 29.26, Rsk – 0.33, Wa – 83.81 ± 29.55 nm). Several deviations in microgeometry of the cilia form factor were revealed, which associated with formation of abnormally long cilia (10–12 μm), decreasing (0.095–0.15 μm) and/or a thickening (0.3–0.4 μm) of their diameter, as well as spatial disorientation like the “corkscrew twisting”. Based on the electron microscopic analysis, anomalies in external dynein arms of the cilia axoneme were revealed, which made it possible to confirmed in two patients the hereditary respiratory pathology.

A Novel Major Pilin Subunit Protein FimM Is Involved in Adhesion of Bifidobacterium longum BBMN68 to Intestinal Epithelial Cells.

Adhesion to the gastrointestinal tract is considered to be important for bifidobacteria to colonize the human gut and exert their probiotic effects. Some cell surface proteins of bifidobacteria, known as adhesins, play critical roles in the binding to host cells or the extracellular matrix (ECM). To elucidate the mechanisms associated with the adhesion of Bifidobacterium longum BBMN68, a centenarian originated potential probiotic, PSORTdb was employed to identify putative extracellular localized proteins in the B. longum BBMN68. Of the 560 predicted extracellular proteins, 21 were further identified as putative adhesion proteins using the conserved domain database of NCBI, and four were successfully overexpressed in the heterologous host, Lactococcus lactis NZ9000. Notably, a recombinant strain expressing FimM showed a significantly increased adhesive affinity for both HT-29 and mucus-secreting LS174T goblet cells (2.2- and 5.4-fold higher than that of the control strain, respectively). Amino acid sequence alignment showed that FimM is a major pilin subunit protein containing a Cna-B type domain and a C-terminal LPKTG sequence. However, in silico analysis of the fimM-coding cluster revealed that BBMN68_RS10200, encoding a pilus-specific class C sortase, was a pseudogene, indicating that FimM may function as a surface adhesin that cannot polymerize into a pili-like structure. Immunogold electron microscopy results further confirmed that FimM localized to the surface of L. lactis NZfimM and B. longum BBMN68 but did not assemble into pilus filaments. Moreover, the adhesive affinity of L. lactis NZfimM to fibronectin, fibrinogen, and mucin were 3.8-, 2.1-, and 3.1-fold higher than that of the control. The affinity of FimM for its attachment receptors was further verified through an inhibition assay using anti-FimM antibodies. In addition, homologs of FimM were found in Bifidobacterium bifidum 85B, Bifidobacterium gallinarum CACC 514, and 23 other B. longum strains by sequence similarity analysis using BLASTP. Our results suggested that FimM is a novel surface adhesin that is mainly present in B. longum strains.

CrebA increases secretory capacity through direct transcriptional regulation of the secretory machinery, a subset of secretory cargo, and other key regulators.

Specialization of many cells, including the acinar cells of the salivary glands and pancreas, milk-producing cells of mammary glands, mucus-secreting goblet cells, antibody-producing plasma cells, and cells that generate the dense extracellular matrices of bone and cartilage, requires scaling up both secretory machinery and cell-type specific secretory cargo. Using tissue-specific genome-scale analyses, we determine how increases in secretory capacity are coordinated with increases in secretory load in the Drosophila salivary gland (SG), an ideal model for gaining mechanistic insight into the functional specialization of secretory organs. Our findings show that CrebA, a bZIP transcription factor, directly binds genes encoding the core secretory machinery, including protein components of the signal recognition particle and receptor, ER cargo translocators, Cop I and Cop II vesicles, as well as the structural proteins and enzymes of these organelles. CrebA directly binds a subset of SG cargo genes and CrebA binds and boosts expression of Sage, a SG-specific transcription factor essential for cargo expression. To further enhance secretory output, CrebA binds and activates Xbp1 and Tudor-SN. Thus, CrebA directly upregulates the machinery of secretion and additional factors to increase overall secretory capacity in professional secretory cells; concomitant increases in cargo are achieved both directly and indirectly.

Bacteroides fragilis prevents Salmonella Heidelberg translocation in co-culture model mimicking intestinal epithelium

Salmonella Heidelberg is one of the most common serovar causing foodborne illnesses. To limit the development of digestive bacterial infection, food supplements containing probiotic bacteria can be proposed. Commensal non-toxigenic Bacteroides fragilis has recently been suggested as a next-generation probiotic candidate. By using an original triple co-culture model including Caco-2 cells (representing human enterocytes), HT29-MTX (representing mucus-secreting goblet cells), and M cells differentiated from Caco-2 by addition of Raji B lymphocytes, bacterial translocation was evaluated. The data showed that S. Heidelberg could translocate in the triple co-culture model with high efficiency, whereas for B. fragilis a weak translocation was obtained. When cells were exposed to both bacteria, S. Heidelberg translocation was inhibited. The cell-free supernatant of B. fragilis also inhibited S. Heidelberg translocation without impacting epithelial barrier integrity. This supernatant did not affect the growth of S. Heidelberg. The non-toxigenic B. fragilis confers health benefits to the host by reducting bacterial translocation. These results suggested that the multicellular model provides an efficient in vitro model to evaluate the translocation of pathogens and to screen for probiotics that have a potential inhibitory effect on this translocation.

Alpha-tocopherol exerts protective function against the mucotoxicity of particulate matter in amphibian and human goblet cells

Exposure to particulate matter (PM) in ambient air is known to increase the risk of cardiovascular disorders and mortality. The cytotoxicity of PM is mainly due to the abnormal increase of reactive oxygen species (ROS), which damage cellular components such as DNA, RNA, and proteins. The correlation between PM exposure and human disorders, including mortality, is based on long-term exposure. In this study we have investigated acute responses of mucus-secreting goblet cells upon exposure to PM derived from a heavy diesel engine. To this end, we employed the mucociliary epithelium of amphibian embryos and human Calu-3 cells to examine PM mucotoxicity. Our data suggest that acute exposure to PM significantly impairs mucus secretion and results in the accumulation of mucus vesicles in the cytoplasm of goblet cells. RNA-seq analysis revealed that acute responses to PM exposure significantly altered gene expression patterns; however, known regulators of mucus production and the secretory pathway were not significantly altered. Interestingly, pretreatment with α-tocopherol nearly recovered the hyposecretion of mucus from both amphibian and human goblet cells. We believe this study demonstrates the mucotoxicity of PM and the protective function of α-tocopherol on mucotoxicity caused by acute PM exposure from heavy diesel engines.

Tributyrin differentially regulates inflammatory markers and modulates goblet cells number along the intestinal tract segments of weaning pigs

Butyric acid is widely used in pig production as feed additive to improve growth performance at weaning, based on its intestinal health-promoting action. The aim of this study was to evaluate the intestinal architecture and expression of inflammatory cytokines and tight junctions (TJ) markers in weaning piglets fed with tributyrin, either free or microencapsulated, as dietary source of butyric acid. One hundred and eight weaned piglets were divided into pens (4 piglets/pen, n = 9) and received either a basal diet (control) or the basal diet supplemented with tributyrin at 1750 mg/kg (f-TB) or its encapsulated form (m-TB) at 3800 mg/kg (providing 1750 mg/kg of tributyrin equally to f-TB). Growth performance was recorded until d21 when 7 pigs/group were euthanized to collect intestinal samples (duodenum, jejunum, ileum, colon) for histomorphology, cytokines, and TJ markers gene expression analysis. Data were analyzed with 1-way ANOVA. Growth performance was not affected. Compared with control pigs, m-TB pigs tended to induce deeper crypts in the duodenum (P = 0.09); goblet cell number tended to be reduced by f-TB treatment in duodenum villi (P = 0.09), by both f-TB and m-TB treatments in ileum villi (P = 0.06), and by m-TB treatment in colon crypts (P = 0.08). Compared with control animals, f-TB pigs showed higher tumor necrosis factor α (TNF-α) expression in duodenum (trend with P = 0.06), and higher interferon γ (IFN-γ) in the jejunum (P = 0.04), whereas in the colon m-TB pigs tended to down regulate IFN-γ (P = 0.06). Claudin-1 mRNA in the jejunum was reduced in m-TB group compared to control group (P = 0.05). Occludin mRNA in the ileum was reduced in both groups treated with tributyrin compared to control group (P < 0.01). In the colon occludin mRNA was downregulated both in f-TB and m-TB pigs compared to control pigs (P < 0.05).Overall, the supplementation of tributyrin in the diet reduced the mucous-secreting goblet cells in a tract-specific way and modulated the expression of inflammatory cytokines and TJ components along the intestinal tract of weaning piglets, without any negative effect on growth performance.

Proteomic profiling of yellow catfish (Pelteobagrus fulvidraco) skin mucus identifies differentially-expressed proteins in response to Edwardsiella ictaluri infection

Fish mucus acts as a physiological and immunological barrier for maintaining normal fish physiology and conferring defense against pathogens infection. Here we report proteomic profiling of skin mucus of yellow catfish before and after E. ictaluri infection by Label-free LC-MS/MS approach. A total of 918 non-redundant proteins were identified from 54443 spectra referring to yellow catfish genome database. Further annotation via GO and KEGG database revealed complex protein composition of yellow catfish mucus. Besides structural proteins in mucus, a lot of immune-related proteins were retrieved, such as lectins, complement components, antibacterial peptides and immunoglobins. 133 differentially-expressed proteins (DEPs), including 76 up-regulated and 57 down-regulated proteins, were identified, most of which were enriched into 17 pathways centering on “immune system” category with 33 proteins involved. Consistently, significant proliferation of mucus-secreting goblet cells and CYPA-expressing cells were observed along outside of yellow catfish skin after E. ictaluri infection, indicating an enhanced immune response to E. ictaluri infection in yellow catfish skin mucus. The proteomic data provide systematic protein information to comprehensively understand the biological function of yellow catfish skin mucus in response to bacterial infection.