RESUMEN

This article investigates an endoscopic approach that utilizes negative pressure to achieve laser-induced thermal coagulation limited to the esophageal wall's mucosal and superficial submucosal layers. The study was built upon a series of studies combining numerical simulation based on the Monte-Carlo technique and ex vivo porcine tissue experiments, including apparatus design and histology analysis. An endoscopy apparatus was developed using 3D printing to validate the tissue stretching-based approach. A fiber-pigtailed diode was used as the near-infrared source, emitting 208.8 W/cm2 laser irradiance at 1.5 µm. Simulation results suggested that the approach successfully created a local heat well to prevent residual thermal effects (>65°C) from penetrating the deeper submucosal layer. Histology analysis of ex vivo tissues showed that at a fluence of 5.22 kJ/cm2, the depth of thermal coagulation was reduced by half compared to the control. With further preclinical studies, including endoscopy apparatus design, the approach can be applied to the larger esophageal surface.

Asunto(s)

Terapia por Láser , Animales , Porcinos , Endoscopía , Rayos Láser , Luz , Fototerapia

RESUMEN

Carotenoids have been related to a number of health benefits. Their dietary intake and circulating levels have been associated with a reduced incidence of obesity, diabetes, certain types of cancer, and even lower total mortality. Their potential interaction with the gut microbiota (GM) has been generally overlooked but may be of relevance, as carotenoids largely bypass absorption in the small intestine and are passed on to the colon, where they appear to be in part degraded into unknown metabolites. These may include apo-carotenoids that may have biological effects because of higher aqueous solubility and higher electrophilicity that could better target transcription factors, i.e., NF-κB, PPARγ, and RAR/RXRs. If absorbed in the colon, they could have both local and systemic effects. Certain microbes that may be supplemented were also reported to produce carotenoids in the colon. Although some bactericidal aspects of carotenoids have been shown in vitro, a few studies have also demonstrated a prebiotic-like effect, resulting in bacterial shifts with health-associated properties. Also, stimulation of IgA could play a role in this respect. Carotenoids may further contribute to mucosal and gut barrier health, such as stabilizing tight junctions. This review highlights potential gut-related health-beneficial effects of carotenoids and emphasizes the current research gaps regarding carotenoid-GM interactions.

Asunto(s)

Carotenoides , Microbioma Gastrointestinal , Humanos , Carotenoides/farmacología , Carotenoides/metabolismo , Colon/metabolismo , Prebióticos , Suplementos Dietéticos

RESUMEN

The microanatomical features of the intestinal tract mucosa layer in different species of tetrapoda vary according to the type of species, tissue, and function of the targeted cells. In the present study, we have evaluated the histological and histochemical variations of the intestinal tract in four species representing superclass tetrapoda. Bufo regularis (toad), Trachylepis quinquetaeniata (lizard), Columba livia domestica (pigeon) and Mus musculus (mouse) were used as representatives for amphibians, reptilians, avians and mammalians respectively. Histologically, the ileum's mucosal layer of the lower tetrapods (toad and lizard) was almost similar and consists of elongated finger-like shape villi lined with simple columnar epithelium and goblet cells. Similarly, the microanatomical features in ileum of higher tetrapod representatives (pigeon and mouse) were characterized by the presence of villi lined with simple columnar epithelium and scattered goblet cells as well as intestinal glands (crypts of Lieberkühn) at the bases of the intestinal villi. In the toad rectum, the mucosal layer was similar to that of the ileum but with shorter villi and more numerous goblet cells. However, the mucosal layer of the rectum in the lizard had low numbers of absorptive columnar epithelial cells with abundant goblet basal cells. Comparatively, the pigeon's rectal mucosa had almost a similar structure to that of ileum but in leaf-like shaped villi. Finally, the rectum of the mouse has narrow rectal pits, instead of villi, lined with goblet cells and absorptive epithelial cells. Histochemically, the ileum in the four studied tetrapod representatives showed varying biodistribution profiles of neutral, sulfated and carboxylated mucins. There are variations encountered in the intestinal brush border and goblet cells of villi in all species as well as the crypts of Lieberkühn in higher tetrapods. Also, the rectum of all tetrapod species showed weak to strong positive signals for the three types of mucins in the brush border and goblet cells of villi in all species and crypts of Lieberkühn in higher tetrapods as well. In addition, the brush border of toad's rectum was lacking sulfated mucins and that of the lizard did not have any type of mucins. The data of this study will contribute to understand the relationship between the microanatomical features and mucins biodistribution profiles in the mucosal layer of tetrapod intestinal tract and their functions.

Asunto(s)

Lagartos , Mucinas , Animales , Columbidae/metabolismo , Mucosa Intestinal/metabolismo , Intestinos , Lagartos/metabolismo , Mamíferos/metabolismo , Ratones , Mucinas/metabolismo , Distribución Tisular

RESUMEN

The microanatomical features of the oesophageal gastric tract in tetrapod representatives and their function, especially those related to the mucosal layer, have not yet been fully investigated. The mucosal layer cells and their function in the oesophageal gastric tract differ structurally and functionally in tetrapod representatives based on interspecies difference and the type of food and feeding habits. The present study was, therefore, postulated to compare the mucosal microanatomical structure and histochemical biodistribution of different mucin types in oesophageal gastric tract tissues of four tetrapod species. A representative of each tetrapod class was selected, as follows: the Egyptian toad Bufo regularis, the lizard Trachylepis quinquetaeniata, the domestic pigeon Columba livia domestica and the albino mouse Mus musculus for Amphibia, Reptilia, Aves and Mammalia, respectively. Microanatomically, in lower tetrapods (toad and lizard), the mucosal layer of the oesophagus was composed of simple ciliated columnar epithelium with goblet cells, whereas in higher tetrapods (pigeon and mouse) it was composed of stratified squamous epithelium, with non-keratinised epithelium in the pigeon but keratinised epithelium in the mouse. However, the gastric mucosal layer of the stomach in lower tetrapods consists of simple columnar epithelium and gastric glands. Similarly, the mucosa of the pigeon's proventriculus consists of simple columnar epithelium with proventricular glands opened into the lumen, whereas mouse mucosa consists of simple columnar epithelium which folds and forms gastric glands with gastric pits having a variety of cell types. Histochemically, the neutral mucin profile biodistribution in the oesophagus mucosal layer was variable. It was strongly positive in the toad and lizard, but was weak in the pigeon and completely negative in the mouse. In contrast it was strongly positive in the gastric mucosa of the toad, lizard and pigeon, but was weak in the mouse's gastric mucosa. On the other hand, the signals of carboxylated and sulfated mucins were found to be different. They were strong in the mucosa of the lizard oesophagus. In contrast, the carboxylated mucins in the gastric mucosa were positive in all representatives except the mouse. The sulfated mucins were, however, seen localised in the mucosal layer cells of the lizard and pigeon only. The study revealed that the microanatomical structures and functions as well as mucin distribution profiles in the oesophageal gastric tract are in line with interspecies difference and the type of food and feeding habits. However, this may need further investigations including more tetrapod representatives.

Asunto(s)

Esófago/química , Mucosa Gástrica/química , Mucinas/metabolismo , Animales , Bufonidae , Columbidae , Esófago/citología , Esófago/metabolismo , Mucosa Gástrica/citología , Mucosa Gástrica/metabolismo , Lagartos , Ratones , Distribución Tisular

RESUMEN

Many aspects of the mechanisms underlying the symbiosis between humans and gut microbes remain unknown and encompass some of the most intriguing questions in microbiome research. An important factor in this symbiosis is the interplay between microbes and human-produced glycans in mucin and breast milk. In this Opinion paper, I propose a synergy between the structural diversity of human mucin glycans and the enzymatic repertoire of the gut microbiome. The contribution of microbes to mucosal health is discussed, and the role of breast milk glycans in mucosal colonization by microbes is explained. The use of prebiotic mucin glycans in general, and specialized infant and medical nutrition in particular, should be considered as the field of interest to modulate the microbiota and improve mucosal health.

Asunto(s)

Microbioma Gastrointestinal , Microbiota , Femenino , Humanos , Lactante , Leche Humana , Mucinas/análisis , Polisacáridos

RESUMEN

Cisplatin is a chemotherapeutic agent widely used for the treatment of solid cancers. Its administration is commonly associated with acute and chronic gastrointestinal dysfunctions, likely related to mucosal and enteric nervous system (ENS) injuries, respectively. Glucagon-like peptide-2 (GLP-2) is a pleiotropic hormone exerting trophic/reparative activities on the intestine, via antiapoptotic and pro-proliferating pathways, to guarantee mucosal integrity, energy absorption and motility. Further, it possesses anti-inflammatory properties. Presently, cisplatin acute and chronic damages and GLP-2 protective effects were investigated in the mouse distal colon using histological, immunohistochemical and biochemical techniques. The mice received cisplatin and the degradation-resistant GLP-2 analog ([Gly2]GLP-2) for 4 weeks. Cisplatin-treated mice showed mucosal damage, inflammation, IL-1ß and IL-10 increase; decreased number of total neurons, ChAT- and nNOS-immunoreactive (IR) neurons; loss of SOX-10-IR cells and reduced expression of GFAP- and S100ß-glial markers in the myenteric plexus. [Gly2]GLP-2 co-treatment partially prevented mucosal damage and counteracted the increase in cytokines and the loss of nNOS-IR and SOX-10-IR cells but not that of ChAT-IR neurons. Our data demonstrate that cisplatin causes mucosal injuries, neuropathy and gliopathy and that [Gly2]GLP-2 prevents these injuries, partially reducing mucosal inflammation and inducing ENS remodeling. Hence, this analog could represent an effective strategy to overcome colonic injures induced by cisplatin.

Asunto(s)

Colon/lesiones , Neoplasias del Colon/tratamiento farmacológico , Sistema Nervioso Entérico/efectos de los fármacos , Péptido 2 Similar al Glucagón/genética , Animales , Colina O-Acetiltransferasa/genética , Cisplatino/efectos adversos , Cisplatino/farmacología , Colon/efectos de los fármacos , Colon/metabolismo , Neoplasias del Colon/complicaciones , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Modelos Animales de Enfermedad , Sistema Nervioso Entérico/metabolismo , Sistema Nervioso Entérico/patología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interleucina-10/genética , Interleucina-1beta/genética , Ratones , Neuroglía/efectos de los fármacos , Neuroglía/patología , Neuronas/efectos de los fármacos , Neuronas/patología , Óxido Nítrico Sintasa de Tipo II/genética

RESUMEN

BACKGROUND: The mucosal layer plays an important role in regulating the intestinal barrier function. However, the underlying mechanisms of intestinal barrier dysfunction caused by trauma-hemorrhagic shock (THS) are still unknown. METHODS: In this study, we examined the barrier damages, inflammatory responses as well as the metabolic changes of the mucosal layer of the colon in a THS rat model. RESULTS: The results showed that compared to the rats treated with trauma only, THS induced marked failure of intestinal barrier characterized by increased intestinal permeability, inflammatory cell infiltration and decreased expression of genes involved in epithelial integrity. Moreover, decreased colonic mucus content and goblet cell numbers indicated that the mucosal layer was also impaired in response to THS. This was companied by the anomalous inflammatory responses in the tissue. Finally, microdialysis catheter examination showed that metabolites including glycerol, glucose, lactate and pyruvate, glutamate and glutamine were also altered by THS. CONCLUSION: Our results provide evidence that mucus layer-associated metabolic changes may contribute to the THS-induced intestinal barrier dysfunction.

Asunto(s)

Mucosa Intestinal/metabolismo , Choque Hemorrágico/metabolismo , Heridas y Lesiones/metabolismo , Animales , Mucosa Intestinal/patología , Masculino , Insuficiencia Multiorgánica/etiología , Insuficiencia Multiorgánica/metabolismo , Insuficiencia Multiorgánica/fisiopatología , Permeabilidad , Ratas , Ratas Sprague-Dawley , Choque Hemorrágico/patología , Heridas y Lesiones/patología

RESUMEN

OBJECTIVE: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. METHODS: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. RESULTS: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, gga-miR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, gga-miR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. CONCLUSION: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.

RESUMEN

Necrotic enteritis (NE) is a serious problem to the poultry farms, which report NE outbreaks more than once per year, as a result of the inappropriate use of antibiotics in the feed. The NE affected bird die rapidly as a result of various pathophysiological complications in the intestine and immune system. Also, several studies have reported that the genes exclusively related to intestine and immune functions are significantly altered in response to NE. In this study, NE was induced in two genetically disparate chicken lines that are resistant (line 6.3) and sensitive (line 7.2) to avian leukosis and Marek's disease. The intestinal mucosal layer was collected from NE-induced and control chickens, and subjected to RNA-sequencing analysis. The involvement of differentially expressed genes in the intestinal mucosal layer of line 6.3 and 7.2 with the immune system-related pathways was investigated. Among the identified immune system-related pathways, a candidate pathway known as chicken cytosolic DNA-sensing pathway (CDS pathway) was selected for further investigation. RNA-sequencing and pathway analysis identified a total of 21 genes that were involved in CDS pathway and differentially expressed in the intestinal mucosal layer of lines 6.3 and 7.2. The expression of CDS pathway genes was further confirmed by real-time qPCR. In the results, a majority of the CDS pathway genes were significantly altered in the NE-induced intestinal mucosal layer from lines 6.3 and 7.2. In conclusion, our study indicate that NE seriously affects several genes involved in innate immune defense and foreign DNA sensing mechanisms in the chicken intestinal mucosal layer. Identifying the immune genes affected by NE could be an important evidence for the protective immune response to NE-causative pathogens.

Asunto(s)

Pollos , ADN/inmunología , Enteritis/veterinaria , Regulación de la Expresión Génica , Inmunidad Mucosa/genética , Mucosa Intestinal/inmunología , Enfermedades de las Aves de Corral/genética , Animales , Pollos/genética , Pollos/inmunología , Clostridium perfringens , Eimeria , Enteritis/genética , Enteritis/inmunología , Enteritis/microbiología , Mucosa Intestinal/metabolismo , Necrosis , Enfermedades de las Aves de Corral/inmunología , Análisis de Secuencia de ARN

RESUMEN

The gut epithelial barrier, which is composed of the mucosal layer and the intestinal epithelium, has multiple defense mechanisms and interconnected regulatory mechanisms against enteric microbial pathogens. However, many bacterial pathogens have highly evolved infectious stratagems that manipulate mucin production, epithelial cell-cell junctions, cell death, and cell turnover to promote their replication and pathogenicity in the gut epithelial barrier. In this review, we focus on current knowledge about how bacterial pathogens regulate mucin levels to circumvent the epithelial mucus barrier and target cell-cell junctions to invade deeper tissues and increase their colonization. We also describe how bacterial pathogens manipulate various modes of epithelial cell death to facilitate bacterial dissemination and virulence effects. Finally, we discuss recent investigating how bacterial pathogens regulate epithelial cell turnover and intestinal stem cell populations to modulate intestinal epithelium homeostasis.

Asunto(s)

Colon , Mecanismos de Defensa , Células Epiteliales , Homeostasis , Uniones Intercelulares , Mucosa Intestinal , Mucinas , Moco , Células Madre , Uniones Estrechas , Virulencia

RESUMEN

The antiviral effect of a catalytic RNA-hydrolyzing antibody, 3D8 scFv, for intranasal administration against avian influenza virus (H1N1) was described. The recombinant 3D8 scFv protein prevented BALB/c mice against H1N1 influenza virus infection by degradation of the viral RNA genome through its intrinsic RNA-hydrolyzing activity. Intranasal administration of 3D8 scFv (50 µg/day) for five days prior to infection demonstrated an antiviral activity (70% survival) against H1N1 infection. The antiviral ability of 3D8 scFv to penetrate into epithelial cells from bronchial cavity via the respiratory mucosal layer was confirmed by immunohistochemistry, qRT-PCR, and histopathological examination. The antiviral activity of 3D8 scFv against H1N1 virus infection was not due to host immune cytokines or chemokines, but rather to direct antiviral RNA-hydrolyzing activity of 3D8 scFv against the viral RNA genome. Taken together, our results suggest that the RNase activity of 3D8 scFv, coupled with its ability to penetrate epithelial cells through the respiratory mucosal layer, directly prevents H1N1 virus infection in a mouse model system.

Asunto(s)

Anticuerpos Catalíticos/administración & dosificación , Antivirales/administración & dosificación , Células Epiteliales/inmunología , Subtipo H1N1 del Virus de la Influenza A/inmunología , Infecciones por Orthomyxoviridae/prevención & control , Ribonucleasas/administración & dosificación , Anticuerpos de Cadena Única/administración & dosificación , Administración Intranasal , Animales , Antivirales/farmacocinética , Hidrólisis , Ratones Endogámicos BALB C , ARN Viral/metabolismo , Anticuerpos de Cadena Única/farmacocinética , Resultado del Tratamiento

RESUMEN

The Na⁺/H⁺ exchanger NHE8 is expressed on the apical membrane of intestinal epithelial cells and is particularly abundant in the colon. Our previous study showed that Muc2 expression was significantly reduced in NHE8-knockout (NHE8-/-) mice, suggesting that NHE8 plays a role in mucosal protection in the colon. The current study confirms and extends our studies on the role of NHE8 in mucosal protection. The number of bacteria attached on the distal colon was significantly increased in NHE8-/- mice compared with their wild-type littermates. As expected, IL-4 expression was markedly increased in NHE8-/- mice compared with wild-type mice. Immunohistochemistry showed disorganization in the mucin layer of NHE8-/- mice, suggesting a possible direct bacteria-epithelia interaction. Furthermore, NHE8-/- mice were susceptible to dextran sodium sulfate-induced mucosal injury. In wild-type mice, dextran sodium sulfate treatment inhibited colonic NHE8 expression. In Caco-2 cells, the absence of NHE8 expression resulted in higher adhesion rates of Salmonella typhimurium but not Lactobacillus plantarum. Similarly, in vivo, S. typhimurium adhesion rate was increased in NHE8-/- mice compared with wild-type mice. Our study suggests that NHE8 plays important roles in protecting intestinal epithelia from infectious bacterial adherence.

Asunto(s)

Adhesión Bacteriana/fisiología , Inmunidad Mucosa/fisiología , Mucosa Intestinal/metabolismo , Salmonella typhimurium/metabolismo , Intercambiadores de Sodio-Hidrógeno/metabolismo , Animales , Bacterias/clasificación , Células CACO-2 , Colitis/inducido químicamente , Colitis/metabolismo , Colon/metabolismo , Colon/microbiología , Citocinas/metabolismo , Sulfato de Dextran , Regulación de la Expresión Génica/inmunología , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Mucosa Intestinal/microbiología , Masculino , Ratones , Ratones Noqueados , Salmonelosis Animal/inmunología , Salmonelosis Animal/microbiología , Intercambiadores de Sodio-Hidrógeno/genética , Factores de Tiempo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Intestinal mucosal layers are colonized by a complex microbiota that provides beneficial effects under normal physiological conditions, but is capable of contributing to chronic inflammatory disease such as inflammatory bowel disease (IBD) in susceptible individuals. Studies have shown that the enteric microbiota may drive the development of the gut immune system and can induce immune homeostasis as well as contribute to the development of IBD although the precise etiology is still unknown. Therefore, intestinal microbes seem to play a key role in the disease pathogenesis. Especially, dysbiosis, which is a shift in the composition of enteric microbiota to a nonphysiologic composition, is associated with one or more defects in mucosal immune functions, including microbe recognition, barrier function, intercellular communication, and anti-microbial effector mechanisms. This review focuses on the impact of enteric microbiota on the development and perpetuation of IBD. In addition, interactions with enteric bacteria and mucosal cells, including intestinal epithelial cells, dendritic cells, and T cells, to induce immune responses at mucosal surfaces have been discussed in the point of IBD pathogenesis. Further extension of the knowledge of enteric microbiota may lead to insights on the pathogenesis and new therapeutic strategies for IBD.

Asunto(s)

Humanos , Fenómenos Fisiológicos Bacterianos , Interacciones Huésped-Patógeno , Enfermedades Inflamatorias del Intestino/microbiología , Mucosa Intestinal/inmunología , Intestinos/microbiología , Linfocitos T/inmunología