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This study investigates Cystobasidium benthicum (Cb) probiotic yeast and Cyrtocarpa edulis (Ce) fruit dietary effects, single (0.5 %) or combined (Cb:Ce, 0.25:0.25 %), on growth performance, humoral immunity in serum and skin mucus, and intestinal morphology of Nile tilapia (Oreochromis niloticus) after 14 and 28 days. The Cb group presented the highest (P < 0.05) specific growth rate, weight gain, and absolute growth rate with respect to the control group. Immunological assays indicated that Cb, Ce and Cb:Ce groups increased serum nitric oxide concentration compared to the control group (P < 0.05). Cb and Cb:Ce groups showed the highest serum myeloperoxidase enzyme activity at day 14 and 28, respectively (P < 0.05); whereas, Cb:Ce group had the highest (P < 0.05) myeloperoxidase activity in skin mucus. The superoxide dismutase enzyme activity was unaffected. On day 28, Cb, Ce, and Cb:Ce groups showed higher and lower (P < 0.05) catalase enzyme activity in serum and skin mucus, respectively, compared with the control group. Only the Cb group had higher (P < 0.05) total protein concentration in serum (day 14) and skin mucus (day 14 and 28) with respect to the control group. The lysozyme activity in serum (day 28) and skin mucus (day 14) was higher (P < 0.05) in the Cb group compared to the control group. Only the skin mucus of Ce group showed bactericidal activity against Aeromonas dhakensis (P < 0.05). Histological studies indicated that Cb and Cb:Ce groups increased microvilli height, and Cb, Ce and Cb:Ce augmented goblet cell area at day 14 compared to the control group (P < 0.05). At day 28, microvilli height was higher in all groups and the number of intraepithelial leukocytes increased in Cb and Ce groups with respect to the control group (P < 0.05). The ex vivo assay revealed that A. dhakensis in leukocytes decreased cell viability similar to the control group (P < 0.05). A principal component analysis (PCA) confirmed the results. In conclusion, C. benthicum in the diet was the best supplement to improve the growth and immunity of Nile tilapia.
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Alimentación Animal , Cíclidos , Dieta , Frutas , Probióticos , Animales , Probióticos/administración & dosificación , Cíclidos/crecimiento & desarrollo , Cíclidos/inmunología , Dieta/veterinaria , Peroxidasa/metabolismo , Óxido Nítrico/metabolismo , Intestinos/microbiología , Intestinos/inmunología , Piel , Inmunidad Humoral , Moco/metabolismo , Superóxido Dismutasa/metabolismo , Catalasa/metabolismoRESUMEN
The skin of fish is a physicochemical barrier that is characterized by being formed by cells that secrete molecules responsible for the first defense against pathogenic organisms. In this study, the biological activity of peptides from mucus of Seriola lalandi and Seriolella violacea were identified and characterized. To this purpose, peptide extraction was carried out from epidermal mucus samples of juveniles of both species, using chromatographic strategies for purification. Then, the peptide extracts were characterized to obtain the amino acid sequence by mass spectrometry. Using bioinformatics tools for predicting antimicrobial and antioxidant activity, 12 peptides were selected that were chemically produced by simultaneous synthesis using the Fmoc-Tbu strategy. The results revealed that the synthetic peptides presented a random coil or extended secondary structure. The analysis of antimicrobial activity allowed it to be discriminated that four peptides, named by their synthesis code 5065, 5069, 5070, and 5076, had the ability to inhibit the growth of Vibrio anguillarum and affected the copepodite stage of C. rogercresseyi. On the other hand, peptides 5066, 5067, 5070, and 5077 had the highest antioxidant capacity. Finally, peptides 5067, 5069, 5070, and 5076 were the most effective for inducing respiratory burst in fish leukocytes. The analysis of association between composition and biological function revealed that the antimicrobial activity depended on the presence of basic and aromatic amino acids, while the presence of cysteine residues increased the antioxidant activity of the peptides. Additionally, it was observed that those peptides that presented the highest antimicrobial capacity were those that also stimulated respiratory burst in leukocytes. This is the first work that demonstrates the presence of functional peptides in the epidermal mucus of Chilean marine fish, which provide different biological properties when the fish face opportunistic pathogens.
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Acuicultura , Peces , Moco , Animales , Moco/química , Chile , Antioxidantes/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Péptidos/farmacología , Péptidos/química , Péptidos/aislamiento & purificación , Vibrio/efectos de los fármacos , Epidermis/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/química , Antibacterianos/aislamiento & purificaciónRESUMEN
This study introduces the nanobromhexine lipid particle (NBL) platform designed for effective pulmonary drug delivery. Inspired by respiratory virus transport mechanisms, NBL address challenges associated with mucus permeation and inflammation in pulmonary diseases. Composed of low molecular weight polyethylene glycol-coated lipid nanoparticles with bromhexine hydrochloride, NBL exhibit a size of 118 ± 24 nm, a neutral zeta potential, osmolarity of 358 ± 28 mOsmol/kg, and a pH of 6.5. Nebulizing without leakage and showing no toxicity to epithelial cells, NBL display mucoadhesive properties with a 60% mucin-binding efficiency. They effectively traverse the dense mucus layer of Calu-3 cultures in an air-liquid interface, as supported by a 55% decrease in MUC5AC density and a 29% increase in nanoparticles internalization compared to non-exposed cells. In assessing immunomodulatory effects, NBL treatment in SARS-CoV-2-infected lung cells leads to a 40-fold increase in anti-inflammatory MUC1 gene expression, a proportional reduction in pro-inflammatory IL-6 expression, and elevated anti-inflammatory IL-10 expression. These findings suggest a potential mechanism to regulate the excessive IL-6 expression triggered by virus infection. Therefore, the NBL platform demonstrates promising potential for efficient pulmonary drug delivery and immunomodulation, offering a novel approach to addressing mucus permeation and inflammation in pulmonary diseases.
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Pulmón , Nanopartículas , SARS-CoV-2 , Nanopartículas/administración & dosificación , Humanos , Pulmón/metabolismo , SARS-CoV-2/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Inmunomodulación , Línea Celular , Mucina-1/metabolismo , COVID-19 , Lípidos/química , Lípidos/administración & dosificación , Moco/metabolismo , Polietilenglicoles/química , Células Epiteliales/metabolismo , Células Epiteliales/efectos de los fármacos , Tratamiento Farmacológico de COVID-19 , Mucina 5AC/metabolismo , LiposomasRESUMEN
Inflammation and mucus production are prevalent characteristics of chronic respiratory conditions, such as asthma and chronic chronic obstructive pulmonary disease (COPD). Biological co-factors, including bacteria, viruses, and fungi, may exacerbate these diseases by activating various pathways associated with airway diseases. An example is the fungus Pneumocystis, which is linked to severe COPD in human patients. Recent evidence has demonstrated that Pneumocystis significantly enhanced inflammation and mucus hypersecretion in a rat model of elastase-induced COPD. The present study specifically aims to investigate two additional aspects associated with the pathology induced by Pneumocystis infection: inflammation and collagen deposition around airways. To this end, the focus was to investigate the role of the IL-1ß pro-inflammatory pathway during Pneumocystis infection in COPD rats. Several airway pathology-related features, such as inflammation, mucus hypersecretion, and fibrosis, were evaluated using histological and molecular techniques. COPD animals infected with Pneumocystis exhibited elevated inflammation levels, including a synergistic increase in IL-1ß and Cox-2. Furthermore, protein levels of the IL-1ß-dependent transcription factor cAMP response element-binding (CREB) showed a synergistic elevation of their phosphorylated version in the lungs of COPD animals infected with Pneumocystis, while mucus levels were notably higher in the airways of COPD-infected animals. Interestingly, a CREB responsive element (CRE) was identified in the Muc5b promoter. The presence of CREB in the Muc5b promoter was synergistically increased in COPD animals infected with Pneumocystis compared to other experimental groups. Finally, an increment of deposited collagen was identified surrounding the airways of COPD animals infected with Pneumocystis compared with the other experimental animal groups and correlated with the increase of Tgfß1 mRNA levels. These findings emphasize the role of Pneumocystis as a potential biological co-factor in chronic respiratory diseases like COPD or asthma, warranting new perspectives in the treatment of chronic respiratory diseases.
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Asma , Pneumocystis , Neumonía por Pneumocystis , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Ratas , Animales , Elastasa Pancreática/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Pulmón/patología , Asma/metabolismo , Moco/metabolismo , Inflamación/metabolismo , Colágeno/metabolismoRESUMEN
Mucus, produced by Palythoa caribaeorum has been popularly reported due to healing, anti-inflammatory, and analgesic effects. However, biochemical and pharmacological properties of this mucus remains unexplored. Therefore, the present study aimed to study its proteome profile by 2DE electrophoresis and MALDI-TOF. Furthermore, it was evaluated the cytotoxic, antibacterial, and antioxidant activities of the mucus and from its protein extract (PE). Proteomics study identified14 proteins including proteins involved in the process of tissue regeneration and death of tumor cells. The PE exhibited cell viability below 50% in the MCF-7 and S-180 strains. It showed IC50 of 6.9 µg/mL for the J774 lineage, and also, favored the cellular growth of fibroblasts. Furthermore, PE revealed activity against Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Staphylococcus epidermidis (MIC of 250 µg/mL). These findings revealed the mucus produced by Palythoa caribaeorum with biological activities, offering alternative therapies for the treatment of cancer and as a potential antibacterial agent.
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Antozoos , Proteómica , Animales , Antozoos/química , Antibacterianos/farmacología , Proteínas , Moco/microbiología , Pruebas de Sensibilidad MicrobianaRESUMEN
Preterm birth (PB) is a leading cause of perinatal morbidity and mortality. PB prediction is performed by measuring cervical length, with a detection rate of around 70%. Although it is known that a cytokine-mediated inflammatory process is involved in the pathophysiology of PB, none screening method implemented in clinical practice includes cytokine levels as a predictor variable. Here, we quantified cytokines in cervical-vaginal mucus of pregnant women (18-23.6 weeks of gestation) with high or low risk for PB determined by cervical length, also collecting relevant obstetric information. IL-2, IL-6, IFN-γ, IL-4, and IL-10 were significantly higher in the high-risk group, while IL-1ra was lower. Two different models for PB prediction were created using the Random Forest machine-learning algorithm: a full model with 12 clinical variables and cytokine values and the adjusted model, including the most relevant variables-maternal age, IL-2, and cervical length- (detection rate 66 vs. 87%, false positive rate 12 vs. 3.33%, false negative rate 28 vs. 6.66%, and area under the curve 0.722 vs. 0.875, respectively). The adjusted model that incorporate cytokines showed a detection rate eight points higher than the gold standard calculator, which may allow us to identify the risk PB risk more accurately and implement strategies for preventive interventions.
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Nacimiento Prematuro , Embarazo , Recién Nacido , Femenino , Humanos , Nacimiento Prematuro/diagnóstico , Citocinas , Interleucina-2 , Vagina , Cuello del Útero , MocoRESUMEN
Tenacibaculosis caused by Tenacibaculum dicentrarchi is the second most important bacterial disease that affects the Chilean salmon industry. The impacted fish show severe external gross skin lesions on different areas of the body. The external mucus layer that covers fish skin contains numerous immune substances that act as one of the main defense barriers against microbial colonization and invasions by potential pathogens. The present in vitro study aimed to evaluate and elucidate the role of the external mucus layer in the susceptibility of Atlantic salmon (Salmo salar) to three Chilean T. dicentrarchi strains and the type strain. For this, mucus collected from healthy and diseased (i.e., with T. dicentrarchi) Atlantic salmon were used, and various antibacterial and inflammatory parameters were analysed. The T. dicentrarchi strains were attracted to the mucus of Atlantic salmon regardless of health status. All four strains adhered to the skin mucus and very quickly grew using the mucus nutrients. Once infection was established, different mucosal defense components were activated in the fish, but the levels of bactericidal activity and of other enzymes were insufficient to eliminate T. dicentrarchi. Alternatively, this pathogen may be able to neutralize or evade these mechanisms. Therefore, the survival of T. dicentrarchi in fish skin mucus could be relevant to facilitate the colonization and subsequent invasion of hosts. The given in vitro results suggest that greater attention should be given to fish skin mucus as a primary defense against T. dicentrarchi.
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Enfermedades de los Peces , Salmo salar , Tenacibaculum , Animales , Piel , Moco , Estado de SaludRESUMEN
Isolated sulfite oxidase (ISOD) and molybdenum cofactor (MoCD) deficiencies are genetic diseases biochemically characterized by the toxic accumulation of sulfite in the tissues of patients, including the brain. Neurological dysfunction and brain abnormalities are commonly observed soon after birth, and some patients also have neuropathological alterations in the prenatal period (in utero). Thus, we investigated the effects of sulfite on redox and mitochondrial homeostasis, as well as signaling proteins in the cerebral cortex of rat pups. One-day-old Wistar rats received an intracerebroventricular administration of sulfite (0.5 µmol/g) or vehicle and were euthanized 30 min after injection. Sulfite administration decreased glutathione levels and glutathione S-transferase activity, and increased heme oxygenase-1 content in vivo in the cerebral cortex. Sulfite also reduced the activities of succinate dehydrogenase, creatine kinase, and respiratory chain complexes II and II-III. Furthermore, sulfite increased the cortical content of ERK1/2 and p38. These findings suggest that redox imbalance and bioenergetic impairment induced by sulfite in the brain are pathomechanisms that may contribute to the neuropathology of newborns with ISOD and MoCD. Sulfite disturbs antioxidant defenses, bioenergetics, and signaling pathways in the cerebral cortex of neonatal rats. CII: complex II; CII-III: complex II-III; CK: creatine kinase; GST: glutathione S-transferase; HO-1: heme oxygenase-1; SDH: succinate dehydrogenase; SO32-: sulfite.
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Corteza Cerebral , Metabolismo Energético , Cofactores de Molibdeno , Sulfito-Oxidasa , Sulfitos , Animales , Ratas , Animales Recién Nacidos , Oxidación-Reducción , Sulfitos/efectos adversos , Sulfito-Oxidasa/metabolismo , Cofactores de Molibdeno/metabolismo , Ratas Wistar , Homeostasis , Mitocondrias/metabolismo , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Antioxidantes/metabolismoRESUMEN
Bovine genital campylobacteriosis (BGC) is a leading cause of return to estrus in cows. The etiologic agent, Campylobacter fetus subsp. venerealis (Cfv) is transmitted by venereal route. Hence, the surrounding reproductive tract microenvironment could play a role in return to estrus in cows. The presence of Cfv in cervicovaginal mucus of Angus breed females provide three experimental groups, which were subject to bacteriome analyses: 10 Cfv-positive cows (CVP), 10 Cfv-negative cows (CVN), and 10 nonsexually active heifers (NSA). Cows with return to estrus showed higher bacterial richness than NSA. Beta diversity analysis showed a significant difference (P = 0.006) in bacterial composition among the three groups analyzed (CVP, CVN, and NSA). However, no significant difference was found when comparing the CVP versus CVN groups. Ureaplasma and Pseudomonas were the genera most frequently observed in NSA, being Ureaplasma the predictor genus to that group, whereas Alistipes, Bacteroides, Rikenellaceae_RC9_gut_group, UCG-005, and UCG-10 were both significantly more abundant and predictors genera in cows with return to estrus. Our results provide an overview of the cervicovaginal bacterial microbiota in cows harboring Cfv and improve the knowledge of the pathogenesis of BGC.
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Infecciones por Campylobacter , Campylobacter fetus , Bovinos , Animales , Femenino , Campylobacter fetus/genética , Infecciones por Campylobacter/veterinaria , Infecciones por Campylobacter/microbiología , MocoRESUMEN
Alveolar macrophages play a crucial role in the initiation and resolution of the immune response in the lungs. Pro-inflammatory M1 alveolar macrophages are an interesting target for treating inflammatory and infectious pulmonary diseases. One commune targeting strategy is to use nanoparticles conjugated with hyaluronic acid, which interact with CD44 overexpressed on the membrane of those cells. Unfortunately, this coating strategy may be countered by the presence on the surface of the nanoparticles of a poly(ethylene glycol) corona employed to improve nanoparticles' diffusion in the lung mucus. This study aims to measure this phenomenon by comparing the behavior in a murine lung inflammation model of three liposomal platforms designed to represent different poly(ethylene glycol) and hyaluronic acid densities (Liposome-PEG, Liposome-PEG-HA and Liposome-HA). In this work, the liposomes were obtained by a one-step ethanol injection method. Their interaction with mucin and targeting ability toward pro-inflammatory macrophages were then investigated in vitro and in vivo in a LPS model of lung inflammation. In vitro, poly(ethylene glycol) free HA-liposomes display a superior targeting efficiency toward M1 macrophages, while the addition of poly(ethylene glycol) induces better mucus mobility. Interestingly in vivo studies revealed that the three liposomes showed distinct cell specificity with alveolar macrophages demonstrating an avidity for poly(ethylene glycol) free HA-liposomes, while neutrophils favored PEGylated liposomes exempt of HA. Those results could be explained by the presence of two forces exercising a balance between mucus penetration and receptor targeting. This study corroborates the importance of considering the site of action and the targeted cells when designing nanoparticles to treat lung diseases.
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Ácido Hialurónico , Liposomas , Ratones , Animales , Macrófagos Alveolares , Polietilenglicoles , MocoRESUMEN
AIMS: This study aimed to evaluate the gastroduodenal protective action of crude fraction extracted from P. caribaeorum mucus in Wistar rats. MAIN METHOD: Initially, phytochemical screening was performed to measure secondary metabolites present in the extract. Subsequently, studies of gastroprotective action in Wistar rats were developed. The animals were randomly divided into six experimental groups: SF0.9% group, misoprostol group, and test groups (200, 100, 10, and 1 mg/kg) that received different doses of the crude fraction of zoanthid mucus (CFZM) diluted in SF0.9%. After 14 days of treatment, acute gastric ulcers were induced by gavage by administering aspirin (200 mg/kg). The stomach and duodenum were removed for histopathological and gene analysis of the mucosa. KEY FINDINGS: The present study found that all investigated metabolites showed negative results. The crude fraction showed a gastric and duodenal protective effect evidenced by an increase in the amount and production of mucins (MUC1 and MUC5AC) and mucus production area in the stomach. Histopathological analysis evidenced a decrease in epithelial damage in the duodenum, with a more significant extension of intestinal villi and a greater amount of goblet cells. SIGNIFICANCE: The crude fraction, extracted from P. caribaeorum, showed gastric and duodenal protective action and is not inert in murine gastroduodenal tissues.
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Antozoos , Úlcera Gástrica , Ratas , Ratones , Animales , Ratas Wistar , Mucosa Gástrica , Moco/metabolismo , Úlcera Gástrica/inducido químicamente , Úlcera Gástrica/tratamiento farmacológico , Úlcera Gástrica/metabolismo , Duodeno/metabolismoRESUMEN
In marine ecosystems, most invertebrates possess diverse microbiomes on their external surfaces, such as those found in the pedal mucus of grazing gastropods and chitons that aids displacement on different surfaces. The microbes are then transported around and placed in contact with free-living microbial communities of micro and other macro-organisms, potentially exchanging species and homogenizing microbial composition and structure among grazer hosts. Here, we characterize the microbiota of the pedal mucus of five distantly related mollusk grazers, quantify differences in microbial community structure, mucus protein and carbohydrate content, and, through a simple laboratory experiment, assess their effects on integrated measures of biofilm abundance. Over 665 Amplicon Sequence Variants (ASVs) were found across grazers, with significant differences in abundance and composition among grazer species and epilithic biofilms. The pulmonate limpet Siphonaria lessonii and the periwinkle Echinolittorina peruviana shared similar microbiota. The microbiota of the chiton Chiton granosus, keyhole limpet Fissurella crassa, and scurrinid limpet Scurria araucana differed markedly from one another, and form those of the pulmonate limpet and periwinkle. Flavobacteriaceae (Bacteroidia) and Colwelliaceae (Gammaproteobacteria) were the most common among microbial taxa. Microbial strict specialists were found in only one grazer species. The pedal mucus pH was similar among grazers, but carbohydrate and protein concentrations differed significantly. Yet, differences in mucus composition were not reflected in microbial community structure. Only the pedal mucus of F. crassa and S. lessonii negatively affected the abundance of photosynthetic microorganisms in the biofilm, demonstrating the specificity of the pedal mucus effects on biofilm communities. Thus, the pedal mucus microbiota are distinct among grazer hosts and can affect and interact non-trophically with the epilithic biofilms on which grazers feed, potentially leading to microbial community coalescence mediated by grazer movement. Further studies are needed to unravel the myriad of non-trophic interactions and their reciprocal impacts between macro- and microbial communities.
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Gammaproteobacteria , Gastrópodos , Microbiota , Poliplacóforos , Animales , Moluscos , Microbiota/genética , Biopelículas , MocoRESUMEN
The study aimed to explore the in vitro digestion of a myoglobin diet and its relationship with the gut microbiota and intestinal barrier at two feeding time points. The in vitro study indicated that myoglobin diets had a higher α-helix content and lower digestibility. The particle sizes of the digested myoglobin diets were higher but their zeta potential values were lower than those of a casein diet. C57BL/6J mice were fed with a casein diet incorporated with 0, 0.38%, 1.13% and 3.39% myoglobin for 3 and 8 weeks. With the increase of the myoglobin content, the relative abundance of Akkermansia muciniphila increased significantly. Moreover, the content of short chain fatty acids tended to increase and then decrease as the myoglobin content increased at 3 weeks. Furthermore, the low-myoglobin diet upregulated the gene expression involved in colonic mucin and tight junction proteins by increasing the proportion of beneficial microbiota. However, the high-myoglobin diet had adverse effects.
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Microbioma Gastrointestinal , Mucosa Intestinal , Mioglobina , Animales , Caseínas/metabolismo , Dieta , Dieta Alta en Grasa , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Ratones , Ratones Endogámicos C57BL , Moco/metabolismo , Mioglobina/administración & dosificación , VerrucomicrobiaRESUMEN
The human colonic mucus is mainly composed of mucins, which are highly glycosylated proteins. The normal commensal colonic microbiota has mucolytic activity and is capable of releasing the monosaccharides contained in mucins, which can then be used as carbon sources by pathogens such as Enterohemorrhagic Escherichia coli (EHEC). EHEC can regulate the expression of some of its virulence factors through environmental sensing of mucus-derived sugars, but its implications regarding its main virulence factor, Shiga toxin type 2 (Stx2), among others, remain unknown. In the present work, we have studied the effects of five of the most abundant mucolytic activity-derived sugars, Fucose (L-Fucose), Galactose (D-Galactose), N-Gal (N-acetyl-galactosamine), NANA (N-Acetyl-Neuraminic Acid) and NAG (N-Acetyl-D-Glucosamine) on EHEC growth, adhesion to epithelial colonic cells (HCT-8), and Stx2 production and translocation across a polarized HCT-8 monolayer. We found that bacterial growth was maximum when using NAG and NANA compared to Galactose, Fucose or N-Gal, and that EHEC adhesion was inhibited regardless of the metabolite used. On the other hand, Stx2 production was enhanced when using NAG and inhibited with the rest of the metabolites, whilst Stx2 translocation was only enhanced when using NANA, and this increase occurred only through the transcellular route. Overall, this study provides insights on the influence of the commensal microbiota on the pathogenicity of E. coli O157:H7, helping to identify favorable intestinal environments for the development of severe disease.
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Escherichia coli Enterohemorrágica , Infecciones por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli , Moco , Escherichia coli Enterohemorrágica/metabolismo , Infecciones por Escherichia coli/microbiología , Escherichia coli O157/metabolismo , Proteínas de Escherichia coli/metabolismo , Expectorantes/metabolismo , Fucosa/metabolismo , Galactosa , Microbioma Gastrointestinal , Humanos , Intestinos/metabolismo , Intestinos/microbiología , Mucinas/metabolismo , Moco/inmunología , Moco/metabolismo , Toxina Shiga II/metabolismo , Virulencia , Factores de Virulencia/metabolismoRESUMEN
Eosinophilic esophagitis (EoE) is a chronic inflammatory condition of the esophagus characterized by increased number of eosinophils. Currently, EoE diagnosis is based on endoscopic procedures for histopathological examination, eosinophils' counting and, often, in clinical practice, the challenge is the differentiation between EoE and gastroesophageal reflux disease (GERD). Our aim was to develop novel peptide ligand to Eosinophil cationic protein (ECP) present in EoE biopsies of patients with potential to be used for detection. We performed a comparative proteomic analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) of esophageal biopsies from pediatric patients with eosinophilic esophagitis, gastroesophageal reflux disease and control individuals. Then, phage display technology was used to select peptides against specific up-regulated protein from EoE patients. Twelve phage clones were selected after three biopanning rounds, and the best phage clone reactivity was evaluated by phage-ELISA assay using esophageal mucus samples from 94 pediatric patients. Mass spectrometry showed that eosinophil cationic protein (ECP) was one of the most up-regulated proteins in EoE patients, which is an eosinophil granule protein usually deposited on tissues to mediate remodeling, but in excess may cause fibrosis and hypertrophy, especially in allergic responses. A highly reactive ECP-ligand peptide (E5) was able to distinguish reactive mucus of EoE patients from GERD and the control individuals by Phage-ELISA, achieving a sensitivity of 84.62%, and a specificity of 82.72%. This is the first study that successfully demonstrated an antibody-like peptide targeting ECP at the esophagus mucus as a useful auxilliary tool for EoE diagnosis with a significant association with atopic disorders and dysphagia.ClinicalTrials.gov no.: NCT03069573.
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Esofagitis Eosinofílica , Reflujo Gastroesofágico , Niño , Cromatografía Liquida , Enteritis , Proteína Catiónica del Eosinófilo , Eosinofilia , Esofagitis Eosinofílica/diagnóstico , Esofagitis Eosinofílica/patología , Eosinófilos/metabolismo , Gastritis , Reflujo Gastroesofágico/complicaciones , Humanos , Ligandos , Moco/metabolismo , Péptidos , Proteómica , Espectrometría de Masas en TándemRESUMEN
Typical enteroaggregative Escherichia coli (tEAEC) is a diarrheagenic E. coli pathotype associated with pediatric and traveler's diarrhea. Even without diarrhea, EAEC infections in children also lead to increased gut inflammation and growth shortfalls. EAEC strain's defining phenotype is the aggregative adherence pattern on epithelial cells attributable to the aggregative adherence fimbriae (AAF). EAEC only causes diarrhea in humans; therefore, not much is known of the exact intestinal region of infection and damage or its interactions with intestinal enterocytes in vivo and in situ. This study aimed to develop a new tEAEC mouse model of infection, characterize the microbiota of infected mice, and evaluate in situ the expression of host adherence and surface molecules triggering EAEC infection and the role of the EAEC AAF-II in adherence. Six-week-old C57BL/6 mice, without previous antibiotic treatment, were orally challenged with EAEC 042 strain or EAEC 042 AAF-II mutant (ΔAAF/II) strain, or DAEC-MXR strain (diffusely adherent E. coli clinical isolate), and with saline solution (control group). Paraffin sections of the colon and ileum were stained with H&E and periodic acid-Schiff. ZO-1, ß-catenin, MUC1, and bacteria were analyzed by immunofluorescence. EAEC-infected mice, in comparison with DAEC-MXR-infected and control mice, significantly lost weight during the first 3 days. After 7 days post-infection, mucus production was increased in the colon and ileum, ZO-1 localization remained unaltered, and morphological alterations were more pronounced in the ileum since increased expression and apical localization of ß-catenin in ileal enterocytes were observed. EAEC-infected mice developed dysbiosis 21 days post-infection. At 4 days post-infection, EAEC strain 042 formed a biofilm on ileal villi and increased the expression and apical localization of ß-catenin in ileal enterocytes; these effects were not seen in animals infected with the 042 ΔAAF/II strain. At 3 days post-infection, MUC1 expression on ileal enterocytes was mainly detectable among infected mice and colocalized with 042 strains on the enterocyte surface. We developed a novel mouse model of EAEC infection, which mimics human infection, not an illness, revealing that EAEC 042 exerts its pathogenic effects in the mouse ileum and causes dysbiosis. This model is a unique tool to unveil early molecular mechanisms of EAEC infection in vivo and in situ.
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Infecciones por Escherichia coli , Íleon , Microbiota , Mucina-1 , beta Catenina , Adhesinas de Escherichia coli/genética , Animales , Adhesión Bacteriana/genética , Diarrea/microbiología , Modelos Animales de Enfermedad , Disbiosis , Escherichia coli/genética , Escherichia coli/metabolismo , Infecciones por Escherichia coli/microbiología , Humanos , Ratones , Ratones Endogámicos C57BL , Mucina-1/genética , Moco/metabolismo , Viaje , beta Catenina/genéticaRESUMEN
We study the dynamics of a binary fluid, where the two fluids are flowing parallel to each other in a cylindrical geometry, and driven by a pulsatile pressure gradient. One of the fluids is a low viscosity Newtonian fluid, the other one is viscoelastic. In order to be able to apply the model to different biofluids, we consider that the viscoelastic fluid has several characteristic times. We characterize the dynamics of the fluids as generalized Darcy's laws, with linear response functions to pulsatile pressure gradients, whose parameters are coupled for both fluids through the fluid-fluid boundary conditions. We apply our results to the dynamics of mucus and air in the trachea and find that the frequency that allows for a larger movement of the mucus, coincides with the experimental frequency of cough. This allows us to propose a plausible explanation for the frequency of cough in healthy individuals, a mechanical process to expel noxious substances from the respiratory system.
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Tos , Tráquea , Humanos , Modelos Biológicos , Moco/fisiología , ViscosidadRESUMEN
This study aimed to evaluate salivary, serum, and abomasal mucus IgA levels in lambs naturally infected with Haemonchus contortus. Thirty-seven crossbred lambs (½ Texel or ½ Ile de France) with an average age of 193 days were evaluated for 56 days after grazing on a contaminated pasture. Fecal samples were collected every 7 days to evaluate the EPG. Blood and saliva samples were collected for IgA measurement every 14 days. On D56, 29 animals were killed for parasite counting and IgA quantification in the abomasal mucus. Salivary, serum, and abomasal mucus IgA were measured by ELISA using third-stage larvae antigens. Salivary and mucus IgA were not correlated, but D14 salivary IgA correlated with EPG on D28 (r = -0.37) and D56 (r = -0.36); D28 salivary IgA correlated with D49 (r = -0.40) and D56 EPG (r = -0.44). Abomasal mucus IgA negatively correlated with EPG from D28 to D56 (r varied from _0.51 to -0.62) and with the counts of all parasitic stages (-0.60 to -0.67). The lambs were classified as susceptible (S) or resistant (R) according to EPG (D56 EPG and cumulative EPG) or IgA (salivary, serum, and mucus IgA). Based on D56 EPG and cumulative EPG, resistant lambs had higher D14 salivary IgA, mucus IgA, and total worm counts. For evaluations based on IgA levels, the EPG of S and R animals differed, indicating that IgA was an immune correlate of protection against natural infection with Haemonchus sp., mainly in the saliva sample of D14.
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Hemoncosis , Haemonchus , Enfermedades de las Ovejas , Animales , Heces/parasitología , Hemoncosis/veterinaria , Inmunoglobulina A , Moco , Recuento de Huevos de Parásitos/veterinaria , Ovinos , Enfermedades de las Ovejas/parasitologíaRESUMEN
Resumen Los tumores neuroendocrinos del colon son relativamente raros en comparación con el carcinoma colorrectal esporádico; su incidencia es muy baja y la localización más frecuente es en el ciego, colon sigmoideo y recto. Se describe con poca frecuencia la asociación de tumor neuroendocrino con la colitis ulcerativa debido a la inflamación crónica que conduce a la diferenciación de células neuroendocrinas a partir de células multipotenciales en el epitelio displásico, que puede ser responsable del desarrollo de carcinomas neuroendocrinos colorrectales. La presentación del caso se trata de un paciente de 57 años de edad con colitis ulcerativa con cuadro de dolor abdominal, pérdida de peso y diarreas líquidas con moco. En el examen físico presentó una lesión endurecida en el marco colónico derecho con tomografía de lesión neoplásica en el colon ascendente y en su hospitalización presentó un cuadro de oclusión intestinal, por lo que se le realizó una hemicolectomía más ileostomía, cuyo resultado en la biopsia fue un tumor neuroendocrino G3 de células grandes en el colon ascendente. En este artículo se revisan de forma práctica los aspectos relacionados con la fisiopatología, diagnóstico y tratamiento de la asociación de estas dos patologías.
Abstract Neuroendocrine colon tumors are relatively unknown compared with sporadic colorectal cancer; its incidence is low, and it is generally located in the cecum, sigmoid colon, and rectum. The existing relation between neuroendocrine tumors and ulcerative colitis is not frequently described due to the chronic inflammation that leads to neuroendocrine cell differentiation from multipotential cells in the dysplastic epithelium, which can be responsible for colorectal neuroendocrine carcinomas development. The study refers to the case of a patient of 57 years old with ulcerative colitis, abdominal pain, weight loss, and liquid diarrhea with mucus. Physical examination revealed a hardened lesion in the right colonic framework with the tomography of a neoplastic lesion in the ascending colon. When the patient was hospitalized, he developed an intestinal obstruction. A hemicolectomy plus ileostomy procedure was performed resulting in a large cell G3 neuroendocrine tumor in the ascending colon revealed in the biopsy. In this article, the aspects related to the pathophysiology, diagnosis, and treatment of the association of these two pathologies are reviewed in a practical way.
Asunto(s)
Humanos , Masculino , Persona de Mediana Edad , Colitis Ulcerosa , Tumores Neuroendocrinos , Colon , Carcinoma Neuroendocrino , Patología , Recto , Terapéutica , Neoplasias Colorrectales , Dolor Abdominal , Colitis , Diarrea , Inflamación , Obstrucción Intestinal , MocoRESUMEN
Caecilians (order Gymnophiona) are apodan, snake-like amphibians, usually with fossorial habits, constituting one of the most unknown groups of terrestrial vertebrates. As in orders Anura (frogs, tree frogs and toads) and Caudata (salamanders and newts), the caecilian skin is rich in mucous glands, responsible for body lubrication, and poison glands, producing varied toxins used in defence against predators and microorganisms. Whereas in anurans and caudatans skin gland morphology has been well studied, caecilian poison glands remain poorly elucidated. Here we characterised the skin gland morphology of the caecilian Siphonops annulatus, emphasising the poison glands in comparison to those of anurans and salamanders. We showed that S. annulatus glands are similar to those of salamanders, consisting of several syncytial compartments full of granules composed of protein material but showing some differentiated apical compartments containing mucus. An unusual structure resembling a mucous gland is frequently observed in lateral/apical position, apparently connected to the main duct. We conclude that the morphology of skin poison glands in caecilians is more similar to salamander glands when compared to anuran glands that show a much-simplified structure.