RESUMEN

The chronic airway infections with Pseudomonas aeruginosa are the major co-morbidity in people with cystic fibrosis (CF). Within CF lungs, P. aeruginosa persists in the conducting airways together with human mucins as the most abundant structural component of its microenvironment. We investigated the adhesion of 41 serial CF airway P. aeruginosa isolates to airway mucin preparations from CF sputa. Mucins and bacteria were retrieved from five modulator-naïve patients with advanced CF lung disease. The P. aeruginosa isolates from CF airways and non-CF reference strains showed a strain-specific signature in their adhesion to ovine, porcine and bovine submaxillary mucins and CF airway mucins ranging from no or low to moderate and strong binding. Serial CF clonal isolates and colony morphotypes from the same sputum sample were as heterogeneous in their affinity to mucin as representatives of other clones thus making 'mucin binding' one of the most variable intraclonal phenotypic traits of P. aeruginosa known to date. Most P. aeruginosa CF airway isolates did not adhere more strongly to CF airway mucins than to plastic surfaces. The strong binders, however, exhibited a strain-specific affinity gradient to O-glycans, CF airway and mammalian submaxillary mucins.

Asunto(s)

Adhesión Bacteriana , Fibrosis Quística , Mucinas , Infecciones por Pseudomonas , Pseudomonas aeruginosa , Esputo , Fibrosis Quística/microbiología , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/aislamiento & purificación , Mucinas/metabolismo , Humanos , Animales , Esputo/microbiología , Infecciones por Pseudomonas/microbiología , Porcinos , Bovinos , Ovinos

RESUMEN

Native mucus is heterogeneous, displays high inter-individual variation and is prone to changes during harvesting and storage. To overcome the lack of reproducibility and availability of native mucus, commercially available purified mucins, porcine gastric mucin (PGM) and mucin from bovine submaxillary gland (BSM), have been widely used. However, the question is to which extent the choice of mucin matters in studies of their interaction with polymers as their composition, structure and hence physicochemical properties differ. Accordingly, the interactions between PGM or BSM with two widely used polymers in drug delivery, polyethylene oxide and chitosan, was studied with orthogonal methods: turbidity, dynamic light scattering, and quartz crystal microbalance with dissipation monitoring. Polymer binding and adsorption to the two commercially available and purified mucins, PGM and BSM, is different depending on the mucin type. PEO, known to interact weakly with mucin, only displayed limited interaction with both mucins as confirmed by all employed methods. In contrast, chitosan was able to bind to both PGM and BSM. Interestingly, the results suggest that chitosan interacts with BSM to a greater extent than with PGM indicating that the choice of mucin, PGM or BSM, can affect the outcome of studies of mucin interactions with polymers.

Asunto(s)

Quitosano , Mucinas Gástricas , Mucinas , Glándula Submandibular , Animales , Bovinos , Porcinos , Quitosano/química , Quitosano/metabolismo , Glándula Submandibular/metabolismo , Glándula Submandibular/química , Mucinas Gástricas/metabolismo , Mucinas Gástricas/química , Mucinas/metabolismo , Mucinas/química , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Polímeros/química , Polímeros/metabolismo , Estómago/química

RESUMEN

Pseudomonas aeruginosa (P. aeruginosa) pneumonia can have serious physiological consequences, particularly when P. aeruginosa biofilms are formed. Although inhaled therapy is preferred, inhaled drugs tend to get trapped by pulmonary mucus, which hinders efficient antibiotic permeability through mucus and biofilms. In this study, we prepare poly[2-(pentamethyleneimino)ethyl methacrylate]-block-poly[2-(N-oxide-pentamethyleneimino)ethyl methacrylate] (PPEMA-b-PPOEMA) micelles loaded with azithromycin (AZM) using reversible addition-fragmentation chain transfer (RAFT) polymerization to achieve effective treatment of P. aeruginosa pneumonia. The zwitterionic structure on the surface of the micelle facilitates the successful traversal of the mucus and optimal concentration within the biofilm. Furthermore, the protonation of piperidine in the polymer enables the micelles to exhibit a positive charge in the acidic environment of a bacterial infection, enhancing AZM's interaction with the bacterium. Both in vivo and in vitro experiments demonstrate that this transmucosal zwitterionic polymer, in combination with a charge reversal strategy, effectively promotes the enrichment of micelles at the site of bacterial infection, thereby increasing the number of antibiotics reaching the bacterial interior and demonstrating remarkable antibacterial synergy. Overall, this work offers a promising approach for trans-airway drug delivery in the treatment of pneumonia.

Asunto(s)

Antibacterianos , Micelas , Pseudomonas aeruginosa , Antibacterianos/química , Antibacterianos/farmacología , Antibacterianos/administración & dosificación , Pseudomonas aeruginosa/efectos de los fármacos , Animales , Mucinas/química , Mucinas/metabolismo , Ratones , Administración por Inhalación , Azitromicina/química , Azitromicina/farmacología , Azitromicina/administración & dosificación , Pruebas de Sensibilidad Microbiana , Infecciones por Pseudomonas/tratamiento farmacológico , Biopelículas/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Neumonía/tratamiento farmacológico , Enfermedad Crónica , Portadores de Fármacos/química

RESUMEN

Ovarian cancer stands as the deadliest gynecologic malignancy, responsible for nearly 65% of all gynecologic cancer-related deaths. The challenges in early detection and diagnosis, coupled with the widespread intraperitoneal spread of cancer cells and resistance to chemotherapy, contribute significantly to the high mortality rate of this disease. Due to the absence of specific symptoms and the lack of effective screening methods, most ovarian cancer cases are diagnosed at advanced stages. While chemotherapy is a common treatment, it often leads to tumor recurrence, necessitating further interventions. In recent years, antibody-drug conjugates (ADCs) have emerged as a valuable tool in targeted cancer therapy. These complex biotherapeutics combine an antibody that specifically targets tumor specific/associated antigen(s) with a high potency anti-cancer drug through a linker, offering a promising approach for ovarian cancer treatment. The identification of molecular targets in various human tumors has paved the way for the development of targeted therapies, with ADCs being at the forefront of this innovation. By delivering cytotoxic agents directly to tumors and metastatic lesions, ADCs show potential in managing chemo-resistant ovarian cancers. Mucins such as MUC16, MUC13, and MUC1 have shown significantly higher expression in ovarian tumors as compared to normal and/or benign samples, thus have become promising targets for ADC generation. While traditional markers are limited by their elevated levels in non-cancerous conditions, mucins offer a new possibility for targeted treatment in ovarian cancer. This review comprehensively described the potential of mucins for the generation of ADC therapy, highlighting their importance in the quest to improve the outcome of ovarian cancer patients.

Asunto(s)

Inmunoconjugados , Mucinas , Neoplasias Ováricas , Humanos , Femenino , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/metabolismo , Inmunoconjugados/uso terapéutico , Inmunoconjugados/farmacología , Mucinas/metabolismo , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacología , Animales

RESUMEN

Central nervous system tuberculosis (CNS-TB) is a severe form of extra-pulmonary tuberculosis with high mortality and morbidity rates. The standard treatment regimen for CNS-TB parallels that of pulmonary TB, despite the challenge posed by the blood-brain barrier (BBB), which limits the efficacy of first-line anti-TB drugs (ATDs). Nose-to-brain (N2B) drug delivery offers a promising solution for achieving high ATD concentrations directly at infection sites in the brain while bypassing the BBB. This study aimed to develop chitosan nanoparticles encapsulating ATDs, specifically isoniazid (INH) and rifampicin (RIF). These nanoparticles were further processed into micro-sized chitosan nano-aggregates (NA) via spray drying. Both INH-NA and RIF-NA showed strong mucoadhesion and significantly higher permeation rates across RPMI 2650 cells compared to free ATDs. Intranasal administration of these NAs to TB-infected mice for four weeks resulted in a significant reduction of mycobacterial load by approximately ∼2.86 Log 10 CFU compared to the untreated group. This preclinical data highlights the efficacy of intranasal chitosan nano-aggregates in treating CNS-TB, demonstrating high therapeutic potential, and addressing brain inflammation challenges. To our knowledge, this study is the first to show nasal delivery of ATD nano-formulations for CNS-TB management.

Asunto(s)

Antituberculosos , Quitosano , Isoniazida , Nanopartículas , Rifampin , Tuberculosis del Sistema Nervioso Central , Animales , Ratones , Tuberculosis del Sistema Nervioso Central/tratamiento farmacológico , Barrera Hematoencefálica , Quitosano/administración & dosificación , Quitosano/química , Nanopartículas/administración & dosificación , Nanopartículas/química , Administración Intranasal , Células Epiteliales/efectos de los fármacos , Antituberculosos/administración & dosificación , Antituberculosos/química , Ratones Endogámicos BALB C , Adhesivos/administración & dosificación , Adhesivos/química , Mucinas/química , Encéfalo/efectos de los fármacos , Encéfalo/patología , Humanos , Línea Celular , Isoniazida/administración & dosificación , Rifampin/administración & dosificación , Sistemas de Liberación de Medicamentos

RESUMEN

Binary systems of citrus peel pectin (a major food carbohydrate) and mucin (a principal oral-gastrointestinal glycoprotein) are studied, as to understand the interactions and thermodynamics between food and biofluids during oral processing and digestion. The fluorimetry emission spectra of mucin were quenched by pectin addition at 293, 301, 310 and 318 K, indicating direct contact between the two macromolecular populations. A red shift, suggesting pectin-induced alterations on mucin conformation, has been observed at 318 K. Intensity-based Stern - Volmer plots fitted second-order polynomial equations, suggesting the coexistence of both static and dynamic quenching, while the increase of the slopes with temperature points to the predominance of dynamic phenomena. Time-resolved fluorescence measurements also point to dynamic quenching related to transient interactions, rather than to specific bonding. Thermodynamic analysis yields negative free energy changes in all cases, with positive changes for enthalpy and large positive values for TΔS. These are in agreement with the Stern - Volmer analysis, suggesting the predominance of transient, dynamic (here entropic) interactions. These provide an image of mucin interacting with pectin macromolecules during the oral processing and digestion of foods, and can relate to the texture, flavor (e.g. astringency) and bioavailability of polysaccharide-based foods.

Asunto(s)

Mucinas , Pectinas , Fluorometría/métodos , Mucinas/química , Mucinas/metabolismo , Pectinas/química , Pectinas/metabolismo , Unión Proteica , Espectrometría de Fluorescencia/métodos , Termodinámica

RESUMEN

Elevated bacterial sialidase activity in the female genital tract is strongly associated with poor health outcomes including preterm birth and bacterial vaginosis (BV). These negative effects may arise from sialidase-mediated degradation of the protective mucus layer in the cervicovaginal environment. Prior biochemical studies of vaginal bacterial sialidases have focused solely on the BV-associated organism Gardnerella vaginalis. Despite their implications for sexual and reproductive health, sialidases from other vaginal bacteria have not been characterized. Here, we show that vaginal Prevotella species produce sialidases that possess variable activity toward mucin substrates. The sequences of sialidase genes and their presence are largely conserved across clades of Prevotella from different geographies, hinting at their importance globally. Finally, we find that Prevotella sialidase genes and transcripts, including those encoding mucin-degrading sialidases from Prevotella timonensis, are highly prevalent and abundant in human vaginal genomes and transcriptomes. Together, our results identify Prevotella as a critical source of sialidases in the vaginal microbiome, improving our understanding of this detrimental bacterial activity.

Asunto(s)

Microbiota , Neuraminidasa , Prevotella , Vagina , Humanos , Prevotella/enzimología , Prevotella/genética , Prevotella/aislamiento & purificación , Neuraminidasa/metabolismo , Neuraminidasa/genética , Femenino , Vagina/microbiología , Mucinas/metabolismo , Vaginosis Bacteriana/microbiología , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética

RESUMEN

Aeromonas salmonicida is an opportunistic pathogen with relevance for aquaculture. Fish epithelia are covered by a mucus layer, composed mainly by highly glycosylated mucins, which are the first point of contact between fish and pathogens. Quorum sensing (QS), a bacterial communication mechanism through secreted autoinducer signals that governs gene expression, influences bacterial growth and virulence. The main A. salmonicida autoinducers are mediated by the luxS and asaI genes, corresponding to inter- and intraspecies communication, respectively. The aim of this study was to determine the effect of the mucins that pathogens encounter during colonization of the gill and skin on A. salmonicida QS. We found that expression of A. salmonicida asaI, but not luxS, was increased after culture at 20 °C compared to 10 °C. Rainbow trout gill and skin mucins up-regulated asaI expression 2-fold but down-regulated luxS 10-fold. The downregulation of luxS was reflected by a reduction in autoinducer-2 secretion. Mucins isolated from skin had a stronger inhibitory effect than mucins isolated from gills on both luxS expression and A1-2 secretion, consistent with a higher relative abundance of N-Acetylneuraminic acid on skin mucins than on gill mucins. Reduction of AI-2 production by mucins or luxS-deletion lead to a reduced A. salmonicida auto-aggregation. Furthermore, after colonization of the gill, luxS was down regulated whereas asaI expression was upregulated. Both in vivo and in vitro, the expression of luxS and asaI were thus differentially regulated, frequently in an inverse manner. The strong AI-2 inhibiting effect of the skin mucins is likely part of the mucin-based defense against pathogens.

Asunto(s)

Aeromonas salmonicida , Homoserina , Mucinas , Oncorhynchus mykiss , Percepción de Quorum , Animales , Oncorhynchus mykiss/inmunología , Aeromonas salmonicida/fisiología , Mucinas/genética , Mucinas/metabolismo , Homoserina/análogos & derivados , Liasas de Carbono-Azufre/genética , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Proteínas Bacterianas/genética , Lactonas , Piel/inmunología , Piel/microbiología , Branquias/inmunología , Branquias/metabolismo

RESUMEN

The crucial role of zwitterionic phosphatidylcholines (PC) within mucus gel is essential for maintaining intestinal homeostasis, while the underlying mechanism remains incompletely understood. Herein, we compared the dynamic interfacial adsorption behavior of saturated dipalmitoylphosphatidylcholine (DPPC) and unsaturated dioleoylphosphatidylcholine (DOPC) to intestinal mucin and their impact on the intestinal mucus barrier function. Results of quartz crystal microbalance with dissipation showed that the highly surface-hydrated DPPC vesicles exhibited significantly faster and more extensive adsorption to purified intestinal mucin than the slightly surface-hydrated DOPC vesicles. Utilizing an intestinal Caco-2/HT29-MTX coculture model, we observed that DPPC vesicles adsorbed much more to the mucus gel compared to DOPC vesicles. Additionally, DPPC vesicle adsorption displayed increased wetting, and converse for DOPC vesicles. Interestingly, both of them exhibited nearly the same protective effects against cell injury induced by peptic-tryptic digests of gliadin (PTG). The partial mechanism involved the binding of PTG to DPPC and DOPC within the mucus gel, thereby restricting PTG contact with the underlying epithelial cells. These findings shed light on the intricate interfacial dynamics of PC adsorption to mucin and their implications for maintaining the integrity of the intestinal mucus barrier.

Asunto(s)

Mucinas , Fosfatidilcolinas , Humanos , Fosfatidilcolinas/química , Adsorción , Mucinas/química , Mucinas/metabolismo , Células CACO-2 , 1,2-Dipalmitoilfosfatidilcolina/química , Mucosa Intestinal/metabolismo , Células HT29 , Propiedades de Superficie , Animales

RESUMEN

Current prophylactic and disease control measures in aquaculture highlight the need of alternative strategies to prevent disease and reduce antibiotic use. Mucus covered mucosal surfaces are the first barriers pathogens encounter. Mucus, which is mainly composed of highly glycosylated mucins, has the potential to contribute to disease prevention if we can strengthen this barrier. Therefore, aim of this study was to develop and characterize fish in vitro mucosal surface models based on commercially available cell lines that are functionally relevant for studies on mucin regulation and host-pathogen interactions. The rainbow trout (Oncorhynchus mykiss) gill epithelial cell line RTgill-W1 and the embryonic cell line from Chinook salmon (Oncorhynchus tshawytscha) CHSE-214 were grown on polycarbonate membrane inserts and chemically treated to differentiate the cells into mucus producing cells. RTGill-W1 and CHSE-214 formed an adherent layer at two weeks post-confluence, which further responded to treatment with the γ-secretase inhibitor DAPT and prolonged culture by increasing the mucin production. Mucins were metabolically labelled with N-azidoacetylgalactosamine 6 h post addition to the in vitro membranes. The level of incorporated label was relatively similar between membranes based on RTgill-W1, while larger interindividual variation was observed among the CHSE in vitro membranes. Furthermore, O-glycomics of RTgill-W1 cell lysates identified three sialylated O-glycans, namely Galß1-3(NeuAcα2-6)GalNAcol, NeuAcα-Galß1-3GalNAcol and NeuAcα-Galß1-3(NeuAcα2-6)GalNAcol, resembling the glycosylation present in rainbow trout gill mucin. These glycans were also present in CHSE-214. Additionally, we demonstrated binding of the fish pathogen A. salmonicida to RTgill-W1 and CHSE-214 cell lysates. Thus, these models have similarities to in vivo mucosal surfaces and can be used to investigate the effect of pathogens and modulatory components on mucin production.

Asunto(s)

Interacciones Huésped-Patógeno , Mucinas , Oncorhynchus mykiss , Animales , Mucinas/metabolismo , Oncorhynchus mykiss/metabolismo , Línea Celular , Membrana Mucosa/metabolismo , Salmón/metabolismo , Branquias/metabolismo , Células Epiteliales/metabolismo , Moco/metabolismo

RESUMEN

Heterotrophic bacteria in the ocean initiate biopolymer degradation using extracellular enzymes that yield low molecular weight hydrolysis products in the environment, or by using a selfish uptake mechanism that retains the hydrolysate for the enzyme-producing cell. The mechanism used affects the availability of hydrolysis products to other bacteria, and thus also potentially the composition and activity of the community. In marine systems, these two mechanisms of substrate processing have been studied in the water column, but to date, have not been investigated in sediments. In surface sediments from an Arctic fjord of Svalbard, we investigated mechanisms of biopolymer hydrolysis using four polysaccharides and mucin, a glycoprotein. Extracellular hydrolysis of all biopolymers was rapid. Moreover, rapid degradation of mucin suggests that it may be a key substrate for benthic microbes. Although selfish uptake is common in ocean waters, only a small fraction (0.5%-2%) of microbes adhering to sediments used this mechanism. Selfish uptake was carried out primarily by Planctomycetota and Verrucomicrobiota. The overall dominance of extracellular hydrolysis in sediments, however, suggests that the bulk of biopolymer processing is carried out by a benthic community relying on the sharing of enzymatic capabilities and scavenging of public goods.

Asunto(s)

Bacterias , Sedimentos Geológicos , Sedimentos Geológicos/microbiología , Biopolímeros/metabolismo , Bacterias/metabolismo , Hidrólisis , Agua de Mar/microbiología , Agua de Mar/química , Polisacáridos/metabolismo , Regiones Árticas , Svalbard , Mucinas/metabolismo

RESUMEN

Sialylation, a crucial post-translational modification of glycoconjugates, entails the attachment of sialic acid (SA) to the terminal glycans of glycoproteins and glycolipids through a tightly regulated enzymatic process involving various enzymes. This review offers a comprehensive exploration of sialylation within the gut, encompassing its involvement in mucosal protection and its impact on disease progression. The sialylation of mucins and epithelial glycoproteins contributes to the integrity of the intestinal mucosal barrier. Furthermore, sialylation regulates immune responses in the gut, shaping interactions among immune cells, as well as their activation and tolerance. Additionally, the gut microbiota and gut-brain axis communication are involved in the role of sialylation in intestinal health. Altered sialylation patterns have been implicated in various intestinal diseases, including inflammatory bowel disease (IBD), colorectal cancer (CRC), and other intestinal disorders. Emerging research underscores sialylation as a promising avenue for diagnostic, prognostic, and therapeutic interventions in intestinal diseases. Potential strategies such as sialic acid supplementation, inhibition of sialidases, immunotherapy targeting sialylated antigens, and modulation of sialyltransferases have been utilized in the treatment of intestinal diseases. Future research directions will focus on elucidating the molecular mechanisms underlying sialylation alterations, identifying sialylation-based biomarkers, and developing targeted interventions for precision medicine approaches.

Asunto(s)

Mucosa Intestinal , Ácido N-Acetilneuramínico , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/inmunología , Animales , Ácido N-Acetilneuramínico/metabolismo , Microbioma Gastrointestinal , Sialiltransferasas/metabolismo , Mucinas/metabolismo , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/inmunología

RESUMEN

The global incidence and prevalence of inflammatory bowel disease (IBD) are gradually increasing. A high-fat diet (HFD) is known to disrupt intestinal homeostasis and aggravate IBD, yet the underlying mechanisms remain largely undefined. Here, a positive correlation between dietary fat intake and disease severity in both IBD patients and murine colitis models is observed. A HFD induces a significant decrease in indole-3-acetic acid (IAA) and leads to intestinal barrier damage. Furthermore, IAA supplementation enhances intestinal mucin sulfation and effectively alleviates colitis. Mechanistically, IAA upregulates key molecules involved in mucin sulfation, including 3'-phosphoadenosine 5'-phosphosulfate synthase 2 (Papss2) and solute carrier family 35 member B3 (Slc35b3), the synthesis enzyme and the transferase of 3'-phosphoadenosine-5'-phosphosulfate (PAPS), via the aryl hydrocarbon receptor (AHR). More importantly, AHR can directly bind to the transcription start site of Papss2. Oral administration of Lactobacillus reuteri, which can produce IAA, contributes to protecting against colitis and promoting mucin sulfation, while the modified L. reuteri strain lacking the iaaM gene (LactobacillusΔiaaM) and the ability to produce IAA fail to exhibit such effects. Overall, IAA enhances intestinal mucin sulfation through the AHR-Papss2-Slc35b3 pathway, contributing to the protection of intestinal homfeostasis.

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A HFD can lead to the development of colitis by disrupting tryptophan metabolism in the gut microbiome and lowering levels of IAA. Supplementation with IAA has been shown to alleviate colitis in mice and improve intestinal barrier function. It is believed that IAA may activate the AHR to upregulate the expression of Papss2 and Slc35b3, promoting sulfation modification of mucins and protecting the intestinal barrier. HFD, high-fat diet; AHR, aryl hydrocarbon receptor; IAA, indole-3-acetic acid; Papss2, 3'-phosphoadenosine 5'-phosphosulfate synthase 2; Slc35b3, solute carrier family 35 member B3.

Asunto(s)

Microbioma Gastrointestinal , Homeostasis , Ácidos Indolacéticos , Mucosa Intestinal , Mucinas , Animales , Humanos , Ratones , Microbioma Gastrointestinal/efectos de los fármacos , Mucinas/metabolismo , Ácidos Indolacéticos/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Ratones Endogámicos C57BL , Colitis/microbiología , Colitis/metabolismo , Colitis/inducido químicamente , Limosilactobacillus reuteri/metabolismo , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/microbiología , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Dieta Alta en Grasa/efectos adversos , Masculino , Receptores de Hidrocarburo de Aril/metabolismo , Receptores de Hidrocarburo de Aril/genética , Modelos Animales de Enfermedad

RESUMEN

AIMS: Ulcerative colitis-associated neoplasia (UCAN) is characterised by multifocal tumourigenesis. A wide range of metachronous lesions have been reported to occur after endoscopic treatment of UCAN, which suggests the development of sporadic tumours in lesions treated as UCAN. Therefore, we aimed to evaluate differences of immunohistochemistry (IHC) in features and clinicopathological characteristics of intramucosal lesions in patients with ulcerative colitis (UC). METHODS AND RESULTS: We examined 35 intramucosal lesions resected for carcinoma or dysplasia by total colectomy from patients with UC and 71 sporadic adenomas (SAs) endoscopically resected from patients without UC. UC lesions were divided into the conventional UCAN group, defined as p53 mutant pattern and normal expression of ß-catenin, and the non-conventional UCAN group, defined as the rest. Ki-67 distribution, α-methylacyl-CoA racemase (AMACR) expression and mucin phenotypes were compared using IHC, and clinicopathological characteristics were investigated. Conventional and non-conventional UCAN lesions were located in the left colon and rectum. Relative to the SA lesions, UCAN lesions occurred in much younger patients and exhibited more frequent basal distribution of Ki-67 in tumour crypts. Conventional UCAN lesions tended to be non-polyploid and exhibited a higher frequency of normal AMACR expression than SA lesions. UC lesions were heterogeneous-only two of the eight patients with multiple lesions had lesions (both non-conventional UCAN lesions) exhibiting concordant IHC staining features. CONCLUSIONS: The basal pattern of Ki-67 distribution, normal expression of AMACR and a non-intestinal mucin phenotype were determined as characteristic features suggestive of UCAN. Non-polypoid growth was another a key feature of UCAN.

Asunto(s)

Colitis Ulcerosa , Antígeno Ki-67 , Mucinas , Racemasas y Epimerasas , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/metabolismo , Colitis Ulcerosa/patología , Colitis Ulcerosa/complicaciones , Neoplasias Asociadas a Colitis/patología , Neoplasias Asociadas a Colitis/etiología , Inmunohistoquímica , Antígeno Ki-67/metabolismo , Mucinas/metabolismo , Fenotipo , Racemasas y Epimerasas/metabolismo

RESUMEN

3D printing can revolutionize personalized medicine by allowing cost-effective, customized tissue-engineering constructs. However, the limited availability and diversity of biopolymeric hydrogels restrict the variety and applications of bioinks. In this study, we introduce a composite bioink for 3D bioprinting, combining a photo-cross-linkable derivative of Mucin (Mu) called Methacrylated Mucin (MuMA) and Hyaluronic acid (HA). The less explored Mucin is responsible for the hydrogel nature of mucus and holds the potential to be used as a bioink material because of its plethora of features. HA, a crucial extracellular matrix component, is mucoadhesive and enhances ink viscosity and printability. Photo-cross-linking with 405 nm light stabilizes the printed scaffolds without damaging cells. Rheological tests reveal shear-thinning behavior, aiding cell protection during printing and improved MuMA bioink viscosity by adding HA. The printed structures exhibited porous behavior conducive to nutrient transport and cell migration. After 4 weeks in phosphate-buffered saline, the scaffolds retain 70% of their mass, highlighting stability. Biocompatibility tests with lung epithelial cells (L-132) confirm cell attachment and growth, suggesting suitability for lung tissue engineering. It is envisioned that the versatility of bioink could lead to significant advancements in lung tissue engineering and various other biomedical applications.

Asunto(s)

Materiales Biocompatibles , Bioimpresión , Ácido Hialurónico , Ensayo de Materiales , Mucinas , Impresión Tridimensional , Ingeniería de Tejidos , Ácido Hialurónico/química , Ácido Hialurónico/farmacología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Humanos , Mucinas/química , Mucinas/metabolismo , Tinta , Luz , Pulmón/citología , Tamaño de la Partícula , Andamios del Tejido/química , Hidrogeles/química , Hidrogeles/farmacología

RESUMEN

Natural polyphenols are promising alternatives to antifungals for novel treatments of vulvovaginal candidiasis (VVC) in an era of antimicrobial resistance. However, polyphenols are poorly soluble and prone to degradation. To overcome their limitations, we propose incorporation in liposomes. The study aimed to develop chitosan and liposome comprising delivery systems for epicatechin (EC) or propyl gallate (PG) as treatment of VVC. EC was selected for its antioxidative properties and PG as an ester of antifungal gallic acid. To improve formulation retention at vaginal site, mucoadhesive chitosan was introduced into formulation as liposomal surface coating or hydrogel due to intrinsic antifungal properties. These polyphenol-loaded liposomes exhibited an average size of 125 nm with a 64 % entrapment efficiency (for both polyphenols). A sustained in vitro polyphenol release was seen from liposomes, particularly in chitosan hydrogel (p < 0.01 or lower). Viscosity was evaluated since increased viscosity upon mucin contact indicated adhesive bond formation between chitosan and mucin confirming mucoadhesiveness of formulations. Antifungal activity was evaluated by the broth microdilution method on Candida albicans CRM-10231. Unlike PG, incorporation of EC in liposomes enabled antifungal activity. Fungicidal activity of chitosan was confirmed both when used as liposomal coating material and as hydrogel vehicle.

Asunto(s)

Antifúngicos , Candida albicans , Candidiasis Vulvovaginal , Catequina , Quitosano , Liposomas , Galato de Propilo , Quitosano/química , Quitosano/administración & dosificación , Antifúngicos/administración & dosificación , Antifúngicos/farmacología , Antifúngicos/química , Candidiasis Vulvovaginal/tratamiento farmacológico , Femenino , Candida albicans/efectos de los fármacos , Catequina/análogos & derivados , Catequina/administración & dosificación , Catequina/química , Catequina/farmacología , Galato de Propilo/administración & dosificación , Galato de Propilo/química , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Hidrogeles/química , Hidrogeles/administración & dosificación , Mucinas/química , Viscosidad , Pruebas de Sensibilidad Microbiana

RESUMEN

Dry eye disease (DED) is a prevalent ocular disorder characterized by unstable tear film condition with loss of aqueous or mucin, excessive oxidative stress, and inflammation, leading to discomfort and potential damage to the ocular surface. Current DED therapies have shown restricted therapeutic effects such as frequent dosing and temporary relief with potential unwanted side effects, urgently necessitating the development of innovative efficient therapeutic approaches. Herein, we developed rosmarinic acid (RosA) conjugated gelatin nanogels loading diquafosol sodium (DQS), DRGNG, for simultaneous ROS-scavenging and mucin-secreting DED treatment. Mechanically, DRGNG suppressed the ROS production, reduced inflammatory factors, and prompted mucin secretion in vitro and in vivo. The whole transcriptome RNA sequencing in vitro further provided a detailed analysis of the upregulation of anti-oxidant, anti-inflammatory, and mucin-promotion pathways. Therapeutically, both in evaporative DED and aqueous deficient DED models, the dual-functional DRGNG could prolong the retention time at the ocular surface, efficiently suppress the oxidative stress response, reverse ocular surface morphology, and recover tear film homeostasis, thus alleviating the DED when the dosage is halved compared to the commercial Diquas®. Our findings contribute to developing innovative therapies for DED and offer insights into the broader applications of nanogels in ocular drug delivery and oxidative stress-related conditions.

Asunto(s)

Cinamatos , Depsidos , Síndromes de Ojo Seco , Gelatina , Nanogeles , Ácido Rosmarínico , Nucleótidos de Uracilo , Depsidos/administración & dosificación , Depsidos/química , Depsidos/farmacología , Síndromes de Ojo Seco/tratamiento farmacológico , Animales , Gelatina/química , Cinamatos/administración & dosificación , Cinamatos/química , Nucleótidos de Uracilo/administración & dosificación , Polifosfatos/química , Humanos , Antioxidantes/administración & dosificación , Antioxidantes/química , Antioxidantes/farmacología , Sistemas de Liberación de Medicamentos , Estrés Oxidativo/efectos de los fármacos , Mucinas/metabolismo , Femenino , Ratones Endogámicos C57BL , Masculino , Especies Reactivas de Oxígeno/metabolismo , Lágrimas/metabolismo , Ratones

RESUMEN

Given that the corneal epithelium is situated on the outermost part of the eye, its functions can be influenced by external temperatures and chemical substances. This study aimed to elucidate the expression profile of chemosensory receptors in corneal epithelial cells and analyze their role in eye function regulation. A comprehensive analysis of 425 chemosensory receptors in human corneal epithelial cells-transformed (HCE-T) revealed the functional expression of TRPV4. The activation of TRPV4 in HCE-T cells significantly increased the expression of membrane-associated mucins MUC1, MUC4, and MUC16, which are crucial for stabilizing tear films, with efficacy comparable to the active components of dry eye medications. The present study suggests that TRPV4, which is activated by body temperature, regulates mucin expression and proposes it as a novel target for dry eye treatment.

Asunto(s)

Epitelio Corneal , Mucina-1 , Mucina 4 , Mucinas , Canales Catiónicos TRPV , Humanos , Antígeno Ca-125/metabolismo , Antígeno Ca-125/genética , Células Epiteliales/metabolismo , Células Epiteliales/citología , Epitelio Corneal/metabolismo , Epitelio Corneal/citología , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/genética , Mucina-1/metabolismo , Mucina-1/genética , Mucina 4/metabolismo , Mucina 4/genética , Mucinas/metabolismo , Mucinas/biosíntesis , Canales Catiónicos TRPV/metabolismo , Canales Catiónicos TRPV/genética

RESUMEN

CONTEXT: The lungs are a common site of tumor metastasis. While morphology and immunophenotype can help differentiate primary from metastatic tumors, distinguishing pulmonary invasive mucinous adenocarcinoma (PIMA) from metastatic colorectal adenocarcinoma (CRC) may occasionally be challenging due to overlapping morphological and immunohistochemical features. Lineage-specific markers such as CDX2, TTF-1, and napsin A are helpful with pulmonary non-mucinous adenocarcinoma (PNMA), however they are non-specific and insensitive when applied to PIMA. SATB2 is a newer marker that distinguishes CRC from upper gastrointestinal and pancreaticobiliary tumors; its utility in distinguishing CRC from PIMA has not been fully elucidated. OBJECTIVE: To evaluate the performance of lineage-specific and mucin glycoprotein immunostains in distinguishing PIMA and CRC. DESIGN: We stained tissue microarrays comprising 34 PNMA, 31 PIMA, and 32 CRC with CK7, CK20, SATB2, CDX2, villin, TTF-1, napsin A, and gel-forming mucins MUC2, MUC5AC, and MUC6. RESULTS: PIMA showed significant (>50% of cells) expression of SATB2 (6%), CDX2 (6%), villin (74%), TTF-1 (13%), and napsin A (23%). However, significant CK7 expression was seen in nearly all PIMA (30/31) and none of the metastatic CRC. CONCLUSION: Our results suggest that CK7 remains one of the most useful markers for distinguishing primary PIMA from metastatic CRC. Expression of the mucin glycoproteins MUC5AC and MUC6 and lack of expression of MUC2 favored a diagnosis of PIMA, but expression of these markers was too heterogeneous to be of clinical utility. To our knowledge this is the only study comparing the immunohistochemical profile of PIMA and metastatic CRC in lung metastasectomy specimens.

Asunto(s)

Adenocarcinoma Mucinoso , Biomarcadores de Tumor , Neoplasias Colorrectales , Inmunohistoquímica , Neoplasias Pulmonares , Humanos , Adenocarcinoma Mucinoso/secundario , Adenocarcinoma Mucinoso/diagnóstico , Adenocarcinoma Mucinoso/patología , Adenocarcinoma Mucinoso/metabolismo , Biomarcadores de Tumor/análisis , Factor de Transcripción CDX2/análisis , Factor de Transcripción CDX2/metabolismo , Neoplasias Colorrectales/patología , Diagnóstico Diferencial , Proteínas de Homeodominio/análisis , Inmunohistoquímica/métodos , Neoplasias Pulmonares/secundario , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/diagnóstico , Proteínas de Unión a la Región de Fijación a la Matriz/análisis , Proteínas de Unión a la Región de Fijación a la Matriz/metabolismo , Proteínas de Microfilamentos/análisis , Mucina 5AC/análisis , Mucina 2/análisis , Mucina 6/análisis , Mucinas/análisis , Mucinas/metabolismo , Análisis de Matrices Tisulares , Factores de Transcripción/análisis

RESUMEN

BACKGROUND: Low-grade appendiceal mucinous neoplasms (LAMNs) are benign non-invasive epithelial proliferations of the appendix. These usually present clinically as mucoceles and these rarely exceed 2 cm in diameter. Lesions confined to the lumen are labelled as LAMN; however those in which mucin spreads outside the peritoneum are labeled as pseudomyxoma peritonei (PMP). AIMS AND OBJECTIVE: A retrospective study was conducted over a period of three years and all cases of appendectomies were studied. Twelve cases of LAMN were identified, which is a diagnostic dilemma for the pathologists and clinicians. RESULTS: LAMN was identified based on the histopathological features. Out of the 12 cases, 9 were classified as LAMN and 3 as appendiceal neoplasm with PMP. There was villous or flat proliferation of epithelial lining, loss lymphoid aggregates, and dissecting mucin within muscularis. CONCLUSION: LAMNs are rare neoplasms of the appendix, with clinical presentation similar to acute appendicitis. Mucinous collections within the appendiceal wall should be extensively searched for mucosal changes and, if found, should prompt a careful search for pushing invasion of LAMNs. A thorough and vigilant gross examination can be of great help. Appendicectomy is the treatment of benign and grossly intact mucinous neoplasm.

Asunto(s)

Adenocarcinoma Mucinoso , Apendicectomía , Neoplasias del Apéndice , Clasificación del Tumor , Seudomixoma Peritoneal , Centros de Atención Terciaria , Humanos , Neoplasias del Apéndice/patología , Neoplasias del Apéndice/cirugía , Masculino , Femenino , Estudios Retrospectivos , Persona de Mediana Edad , Adenocarcinoma Mucinoso/patología , Adenocarcinoma Mucinoso/cirugía , Adenocarcinoma Mucinoso/diagnóstico , Adulto , Seudomixoma Peritoneal/patología , Seudomixoma Peritoneal/cirugía , Seudomixoma Peritoneal/diagnóstico , Anciano , Apéndice/patología , Apéndice/cirugía , Mucinas/metabolismo