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1.
Molecules ; 28(2)2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36677618

RESUMO

UDP-Gal: glycoprotein-N-acetylgalactosamine ß-1,3-galactosyltransferase (T-synthase, EC 2.4.1.122) catalyses the transfer of the monosaccharide galactose from UDP-Gal to GalNAc-Ser/Thr, synthesizing the core 1 mucin type O-glycan. Such glycans play important biological roles in a number of recognition processes. The crucial role of these glycans is acknowledged for mammals, but a lot remains unknown regarding invertebrate and especially mollusc O-glycosylation. Although core O-glycans have been found in snails, no core 1 ß-1,3-galactosyltransferase has been described so far. Here, the sequence of the enzyme was identified by a BlastP search of the NCBI Biomphalaria glabrata database using the human T-synthase sequence (NP_064541.1) as a template. The obtained gene codes for a 388 amino acids long transmembrane protein with two putative N-glycosylation sites. The coding sequence was synthesised and expressed in Sf9 cells. The expression product of the putative enzyme displayed core 1 ß-1,3-galactosyltransferase activity using pNP-α-GalNAc as the substrate. The enzyme showed some sequence homology (49.40% with Homo sapiens, 53.69% with Drosophila melanogaster and 49.14% with Caenorhabditis elegans) and similar biochemical parameters with previously characterized T-synthases from other phyla. In this study we present the identification, expression and characterisation of the UDP-Gal: glycoprotein-N-acetylgalactosamine ß-1,3-galactosyltransferase from the fresh-water snail Biomphalaria glabrata, which is the first cloned T-synthase from mollusc origin.


Assuntos
Biomphalaria , Animais , Humanos , Biomphalaria/genética , Biomphalaria/metabolismo , Sequência de Aminoácidos , Acetilgalactosamina , Drosophila melanogaster/metabolismo , Galactosiltransferases/genética , Galactosiltransferases/química , Polissacarídeos/química , Caenorhabditis elegans/metabolismo , Mucinas , Difosfato de Uridina , Mamíferos/metabolismo
2.
Commun Biol ; 6(1): 85, 2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36690709

RESUMO

Colorectal cancer is a highly heterogeneous disease. Most colorectal cancers are classical adenocarcinoma, and mucinous adenocarcinoma is a unique histological subtype that is known to respond poorly to chemoradiotherapy. The difference in prognosis between mucinous adenocarcinoma and classical adenocarcinoma is controversial. Here, to gain insight into the differences between classical adenocarcinoma and mucinous adenocarcinoma, we analyse 7 surgical tumour samples from 4 classical adenocarcinoma and 3 mucinous adenocarcinoma patients by single-cell RNA sequencing. Our results indicate that mucinous adenocarcinoma cancer cells have goblet cell-like properties, and express high levels of goblet cell markers (REG4, SPINK4, FCGBP and MUC2) compared to classical adenocarcinoma cancer cells. TFF3 is essential for the transcriptional regulation of these molecules, and may cooperate with RPS4X to eventually lead to the mucinous adenocarcinoma mucus phenotype. The observed molecular characteristics may be critical in the specific biological behavior of mucinous adenocarcinoma.


Assuntos
Adenocarcinoma Mucinoso , Adenocarcinoma , Humanos , Mucinas , Adenocarcinoma/patologia , Adenocarcinoma Mucinoso/genética , Adenocarcinoma Mucinoso/patologia , Adenocarcinoma Mucinoso/cirurgia , Prognóstico , Fenótipo , Inibidores de Serinopeptidase do Tipo Kazal/genética
3.
PLoS One ; 18(1): e0273955, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36602978

RESUMO

Lactococcus lactis strains are used as starter cultures in the production of fermented dairy and vegetable foods, but the species also occurs in other niches such as plant material. Lactococcus lactis subsp. lactis G50 (G50) is a plant-derived strain and potential candidate probiotics. Western blotting of cell-wall proteins using antibodies generated against whole G50 cells detected a 120-kDa protein. MALDI-TOF MS analysis identified it as YwfG, a Leu-Pro-any-Thr-Gly cell-wall-anchor-domain-containing protein. Based on a predicted domain structure, a recombinant YwfG variant covering the N-terminal half (aa 28-511) of YwfG (YwfG28-511) was crystallized and the crystal structure was determined. The structure consisted of an L-type lectin domain, a mucin-binding protein domain, and a mucus-binding protein repeat. Recombinant YwfG variants containing combinations of these domains (YwfG28-270, YwfG28-336, YwfG28-511, MubR4) were prepared and their interactions with monosaccharides were examined by isothermal titration calorimetry; the only interaction observed was between YwfG28-270, which contained the L-type lectin domain, and d-mannose. Among four mannobioses, α-1,2-mannobiose had the highest affinity for YwfG28-270 (dissociation constant = 34 µM). YwfG28-270 also interacted with yeast mannoproteins and yeast mannan. Soaking of the crystals of YwfG28-511 with mannose or α-1,2-mannobiose revealed that both sugars bound to the L-type lectin domain in a similar manner, although the presence of the mucin-binding protein domain and the mucus-binding protein repeat within the recombinant protein inhibited the interaction between the L-type lectin domain and mannose residues. Three of the YwfG variants (except MubR4) induced aggregation of yeast cells. Strain G50 also induced aggregation of yeast cells, which was abolished by deletion of ywfG from G50, suggesting that surface YwfG contributes to the interaction with yeast cells. These findings provide new structural and functional insights into the interaction between L. lactis and its ecological niche via binding of the cell-surface protein YwfG with mannose.


Assuntos
Lactococcus lactis , Manose , Manose/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Proteínas de Membrana/metabolismo , Saccharomyces cerevisiae , Lectinas/metabolismo , Mucinas/metabolismo
4.
Chem Pharm Bull (Tokyo) ; 71(1): 70-73, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36596514

RESUMO

In this study, we developed a water-soluble complex-hydrogel viscosity-controlled formulation of amphotericin B (AmB). AmB is insoluble in water, but borax makes it soluble by forming a complex with AmB. Borax also forms complexes with poly(vinyl alcohol) (PVA) to produce viscous hydrogels. Furthermore, boric acid interacts with mucin expressed in corneal epithelial cells. Accordingly, by utilizing these properties of borax simultaneously, we prepared a water-soluble AmB complex-hydrogel with poly(vinyl alcohol)/borate (PVA-B-AmB), which is suitable for eye drops. PVA-B-AmB was easily prepared by simply mixing aqueous AmB solution dissolved in borax, PVA solution, and water. The 11B-NMR results suggested that PVA-B-AmB existed by bonding PVA and AmB via boronic acid. PVA-B-AmB (gel ratio = 0.55) has a viscosity of 18.3 ± 0.5 mPa·s and is suitable for ophthalmic formulations. This formulation exhibited sustained release of AmB of approximately 45% at 24 h. It was also shown that this formulation interacts with mucin. These results suggest that PVA-B-AmB can be used as a water-soluble AmB preparation suitable for ophthalmic use.


Assuntos
Anfotericina B , Hidrogéis , Anfotericina B/química , Álcool de Polivinil/química , Boratos , Mucinas , Água
5.
Artigo em Chinês | MEDLINE | ID: mdl-36631008

RESUMO

Objective To explore how alveolar macrophages from chronic obstructive pulmonary disease (COPD)-model rats affect proliferation and secretion of 16HBE human bronchial epithelial cells and investigate the associated mechanism. Methods Alveolar macrophages were extracted from COPD rats induced by cigarette smoke exposure and LPS instillation through bronchoalveolar lavage, then co-cultured with 16HBE cells in vitro. Exosomes were extracted from alveolar macrophages of rats with exosome isolation kit. The differentially expressed miRNA in exosomes derived from macrophages of rats in COPD group and control group was detected by PCR. miR-380 was overexpressed with miR-380 mimic while the expression of cystic fibrosis transmembrane transduction regulator (CFTR) was knocked down with siRNA in 16HBE cells. The proliferation of 16HBE cells was detected with CCK-8 assay. The migration ability of 16HBE cells was evaluated with TranswellTM migration assay. The levels of mucins (MUC5AC, MUC5B, MUC2) and CFTR expressed by 16HBE cells were detected with Western blot analysis. The expression of TNF-α and IL-6 in the supernatant of 16HBE cells was detected with ELISA. Results The alveolar macrophages from COPD rats enhanced the proliferation and migration of 16HBE cells. The production of mucins and TNF-α as well as IL-6 in 16HBE cells were increased by COPD macrophages. The expression of miR-380 was significantly elevated in exosomes derived from COPD alveolar macrophages. Both overexpression of miR-380 and inhibition of CFTR decreased the expression of CFTR, resulting in the significantly enhanced proliferation and migration of 16HBE cells as well as increased expression of MUC5AC, MUC5B, MUC2 and TNF-α, IL-6. Conclusion The alveolar macrophages from COPD rats can enhance the proliferation and mucin expression as well as inflammatory cytokine secretion of 16HBE cells. This process may be involved with abnormal expression of miR-380 in exosomes of COPD alveolar macrophages and down-regulation of CFTR in bronchial epithelial cells.


Assuntos
MicroRNAs , Doença Pulmonar Obstrutiva Crônica , Ratos , Humanos , Animais , Mucinas/efeitos adversos , Mucinas/metabolismo , Macrófagos Alveolares/metabolismo , Interleucina-6/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/efeitos adversos , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Doença Pulmonar Obstrutiva Crônica/genética , Células Epiteliais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Proliferação de Células
6.
J Agric Food Chem ; 71(1): 700-709, 2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36534057

RESUMO

Reducing sodium intake without decreasing saltiness perception remains an important target in the food industry. This study developed an effective protocol for evaluating the saltiness perception enhanced by umami compounds. Two sodium chloride solutions (2.00 and 6.00 g/L) were the preferred concentrations for consumers. Two-alternative forced-choice evaluation results confirmed that at a concentration of 2.00 g/L (sodium concentration), the highest replacement ratios of monosodium glutamate and l-alanine (Ala) were 10 and 20% in sodium chloride solution without saltiness intensity decrease, respectively. The highest replacement ratios of l-glycine (Gly) and Ala were 10 and 20% compared to 6.00 g/L, respectively. Temporal dominance of sensations analysis figured out that gum Arabic (GA) could compensate for the decrease of the retention time and increase the overall saltiness perception in the sodium-reduced sample. Quartz crystal microbalance with dissipation results showed that Ala and Gly could inhibit the binding of Na+ to mucin, thereby increasing the saltiness perception. GA exhibited the best saltiness enhancement effect in sodium-reduced solution by producing the nanoparticles from GA, decreasing the stability of the solution system, enhancing the loading effect of mucin on Na+, and prolonging the saltiness perception.


Assuntos
Cloreto de Sódio , Paladar , Cloreto de Sódio/análise , Glutamato de Sódio , Sódio , Mucinas
7.
J Theor Biol ; 557: 111334, 2023 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-36306828

RESUMO

The COVID-19 pandemic has underscored the need to understand the dynamics of SARS-CoV-2 respiratory infection and protection provided by the immune response. SARS-CoV-2 infections are characterized by a particularly high viral load, and further by the small number of inhaled virions sufficient to generate a high viral titer in the nasal passage a few days after exposure. SARS-CoV-2 specific antibodies (Ab), induced from vaccines, previous infection, or inhaled monoclonal Ab, have proven effective against SARS-CoV-2 infection. Our goal in this work is to model the protective mechanisms that Ab can provide and to assess the degree of protection from individual and combined mechanisms at different locations in the respiratory tract. Neutralization, in which Ab bind to virion spikes and inhibit them from binding to and infecting target cells, is one widely reported protective mechanism. A second mechanism of Ab protection is muco-trapping, in which Ab crosslink virions to domains on mucin polymers, effectively immobilizing them in the mucus layer. When muco-trapped, the continuous clearance of the mucus barrier by coordinated ciliary propulsion entrains the trapped viral load toward the esophagus to be swallowed. We model and simulate the protection provided by either and both mechanisms at different locations in the respiratory tract, parametrized by the Ab titer and binding-unbinding rates of Ab to viral spikes and mucin domains. Our results illustrate limits in the degree of protection by neutralizing Ab alone, the powerful protection afforded by muco-trapping Ab, and the potential for dual protection by muco-trapping and neutralizing Ab to arrest a SARS-CoV-2 infection. This manuscript was submitted as part of a theme issue on "Modelling COVID-19 and Preparedness for Future Pandemics".


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Pandemias , Anticorpos Antivirais , Sistema Respiratório , Mucinas
8.
J Immunotoxicol ; 20(1): 2148782, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36538286

RESUMO

The Toll-like receptor (TLR) adaptor protein MyD88 is integral to airway inflammatory response to microbial-enriched organic dust extract (ODE) exposures. ODE-induced airway neutrophil influx and release of pro-inflammatory cytokines was essentially abrogated in global MyD88-deficient mice, yet these mice demonstrate an increase in airway epithelial cell mucin expression. To further elucidate the role of MyD88-dependent responses specific to lung airway epithelial cells in response to ODE in vivo, the surfactant protein C protein (SPC) Cre+ embryologic expressing airway epithelial cells floxed for MyD88 to disrupt MyD88 signaling were utilized. The inducible club cell secretory protein (CCSP) Cre+, MyD88 floxed, were also developed. Using an established protocol, mice were intranasally instilled with ODE or saline once or daily up to 3 weeks. Mice with MyD88-deficient SPC+ lung epithelial cells exhibited decreased neutrophil influx following ODE exposure once and repetitively for 1 week without modulation of classic pro-inflammatory mediators including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and neutrophil chemoattractants. This protective response was lost after 3 weeks of repetitive exposure. ODE-induced Muc5ac mucin expression at 1 week was also reduced in MyD88-deficient SPC+ cells. Acute ODE-induced IL-33 was reduced in MyD88-deficient SPC+ cells whereas serum IgE levels were increased at one week. In contrast, mice with inducible MyD88-deficient CCSP+ airway epithelial cells demonstrated no significant difference in experimental indices following ODE exposure. Collectively, these findings suggest that MyD88-dependent signaling targeted to all airway epithelial cells plays an important role in mediating neutrophil influx and mucin production in response to acute organic dust exposures.


Assuntos
Exposição por Inalação , Fator 88 de Diferenciação Mieloide , Animais , Camundongos , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Fator 88 de Diferenciação Mieloide/farmacologia , Exposição por Inalação/efeitos adversos , Transdução de Sinais , Interleucina-6/metabolismo , Receptores Toll-Like , Fator de Necrose Tumoral alfa/metabolismo , Poeira , Mucinas/metabolismo , Mucinas/farmacologia , Camundongos Endogâmicos C57BL
9.
Biomater Adv ; 144: 213235, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36495841

RESUMO

Mucoadhesive thermogels were developed by crosslinking poly(n-isopropylacrylamide) based polymers with chitosan and incorporating disulfide bridges, capable of releasing cysteamine upon interaction with mucin, for the treatment of cystinosis. Through crosslinking with chitosan and incorporating varying concentrations of the disulfide monomer into the polymer backbone, the extent of how mucoadhesive the developed thermogels were could be controlled. Through disulfide bridging with mucin, the thermogels released 6 to 10 µg of the conjugate model 2-mercaptopyridine over five days. Utilizing chitosan as the crosslinker, the developed thermogels were shown to degrade to a statistically higher extent following incubation with lysozyme, the highest concentration tear enzyme, by gravimetric and rheologic analysis. The developed thermogels were extensively tested in vivo utilizing a rat model in which materials were applied directly to the corneal surface and a rabbit model in which thermogels were applied to the inferior fornix. With the developed models, there was no adverse reactions or visual discomfort incurred following application of the thermogels. It has been demonstrated that the thermogels produced can be applied to the inferior fornix and release the stable conjugated payload over several days. The developed thermogel was designed to improve upon the current clinical treatment options for ocular cystinosis which are acidic topical formulations that require reapplication multiple times a day.


Assuntos
Quitosana , Cistinose , Ratos , Animais , Coelhos , Polímeros , Géis , Dissulfetos , Mucinas
10.
ACS Infect Dis ; 9(1): 150-161, 2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36538577

RESUMO

Pseudomonas aeruginosa (P. aeruginosa) is commonly implicated in hospital-acquired infections where its capacity to form biofilms on a variety of surfaces and the resulting enhanced antibiotic resistance seriously limit treatment choices. Because surface attachment sensitizes P. aeruginosa to quorum sensing (QS) and induces virulence through both chemical and mechanical cues, we investigate the effect of surface properties through spatially patterned mucin, combined with sub-inhibitory concentrations of tobramycin on QS and virulence factors in both mucoid and non-mucoid P. aeruginosa strains using multi-modal chemical imaging combining confocal Raman microscopy and matrix-assisted laser desorption/ionization-mass spectrometry. Samples comprise surface-adherent static biofilms at a solid-water interface, supernatant liquid, and pellicle biofilms at an air-water interface at various time points. Although the presence of a sub-inhibitory concentration of tobramycin in the supernatant retards growth and development of static biofilms independent of strain and surface mucin patterning, we observe clear differences in the behavior of mucoid and non-mucoid strains. Quinolone signals in a non-mucoid strain are induced earlier and are influenced by mucin surface patterning to a degree not exhibited in the mucoid strain. Additionally, phenazine virulence factors, such as pyocyanin, are observed in the pellicle biofilms of both mucoid and non-mucoid strains but are not detected in the static biofilms from either strain, highlighting the differences in stress response between pellicle and static biofilms. Differences between mucoid and non-mucoid strains are consistent with their strain-specific phenology, in which the mucoid strain develops highly protected biofilms.


Assuntos
Antibacterianos , Quinolonas , Antibacterianos/farmacologia , Pseudomonas aeruginosa , Quinolonas/farmacologia , Mucinas , Biofilmes , Tobramicina/farmacologia , Fatores de Virulência
11.
Balkan Med J ; 40(1): 57-65, 2023 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-36571426

RESUMO

Background: Allergic rhinitis is a chronic inflammatory disease of the nasal mucosa affecting the quality of life of patients. SRY-box transcription factor 11 (SOX11) was reported to play important roles in inflammatory responses, but its role in AR is poorly understood. Aims: To explore the role of SOX11 in the development of allergic rhinitis. Study Design: Cell culture and animal study. Methods: An in vivo murine allergic rhinitis model was established using ovalbumin treatment in female mice. Interleukin-13-stimulated human nasal mucosa epithelial cells were used for in vitro studies. Expression levels of SOX11, epithelial-derived cytokines, and mucin were determined in both modesls. Results: SOX11 was highly expressed in allergic rhinitis mice. Allergy symptoms, serum ovalbumin-specific IgE, histamine, eosinophils, goblet cells, and type 2 cytokine secretion were increased in ovalbumin-treated mice. Furthermore, allergic rhinitis mice exhibited overproduction of epithelial-derived cytokines (thymic stromal lymphopoietin, interleukin-25, interleukin-33), C-C motif chemokine ligand 26 (CCL26), and mucin 5 AC (MUC5AC). Silencing SOX11 alleviated the behavioral symptoms and upregulation of epithelial-derived cytokines, CCL26, and MUC5AC. In human nasal mucosa epithelial cells, interleukin-13 enhanced SOX11 expression in a time-dependent manner, and signal transducer and activator of transcription 6 (STAT6) was involved in the interleukin-13-mediated expression of SOX11 by regulating transcription. Knockdown of SOX11 reduced epithelial-derived cytokine expression and MUC5AC levels in interleukin-13-treated human nasal mucosa epithelial cells. Conclusion: SOX11 plays a critical role in allergic rhinitis development by regulating epithelial-derived cytokines and might be a new therapeutic target for allergic rhinitis.


Assuntos
Citocinas , Rinite Alérgica , Humanos , Feminino , Camundongos , Animais , Citocinas/metabolismo , Citocinas/uso terapêutico , Interleucina-13/farmacologia , Interleucina-13/uso terapêutico , Ovalbumina/farmacologia , Ovalbumina/uso terapêutico , Qualidade de Vida , Mucinas/uso terapêutico , Fatores de Transcrição SOXC
12.
Biomater Adv ; 145: 213233, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36521413

RESUMO

To achieve and maintain good operability of medical devices while reducing putative side effects for the patient, a promising strategy is to tailor the surface properties of such devices as they critically dictate the tissue compatibility and the biofouling behavior. Indeed, those properties can be strongly improved by generating mucin coatings on such medical devices. However, using coatings on optical systems, e.g., contact lenses, comes with various challenges: here, the geometrical and optical characteristics of the lens may not be compromised by either the coating process or the coating itself. In this study, we show how mucin macromolecules can be attached onto the surfaces of rigid, gas permeable contact lenses while maintaining all critical lens parameters. We demonstrate that the generated coatings improve the surface wettability (contact angles are reduced from 105° to 40° and liquid film break-up times are increased from <1 s to 31 s) and prevent tribological damage to corneal tissue. Additionally, such coatings are highly transparent (transmission values above 98 % compared to an uncoated sample are reached) and efficiently reduce lipid deposition to the lens surface by 90 % but fully maintain the geometrical and mechanical properties of the lenses. Thus, such mucin coatings could also be highly beneficial for other optical systems that are used in direct contact with tissues or body fluids.


Assuntos
Lentes de Contato , Mucinas , Propriedades de Superfície , Molhabilidade , Mucinas/química , Mucinas/farmacologia , Oxigênio , Permeabilidade
13.
Br J Dermatol ; 186(3): 532-543, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34545566

RESUMO

BACKGROUND: The regulation of melanogenesis has been investigated as a long-held aim for pharmaceutical manipulations with denotations for malignancy of melanoma. Mucins have a protective function in epithelial organs; however, in the most outer organ, the skin, the role of mucins has not been studied enough. OBJECTIVES: Our initial hypothesis developed from the identification of correlations between pigmentation and expressions of skin mucins, particularly those existing in skin tissue. We aimed to investigate the action of mucins in human melanocytic cells. MATERIALS AND METHODS: The expression of mucin proteins in human skin was investigated using microarray data from the Human Protein Atlas consortium (HPA) and the Genotype-Tissue Expression consortium (GTEx) database. Mucin expression was measured at RNA and protein levels in melanoma cells. The findings were further validated and confirmed by analysis of independent experiments. RESULTS: We found that the several mucin proteins showed expression in human skin cells and among these, mucin-like protein 1 (MUCL1) showed the highest expression and also clear negative correlation with melanogenesis in epidermal melanocytes. We confirmed the correlations between melanogenesis and MUCL1 by revealing negative correlations in melanocytes with different melanin production, resulting from increased composition of threonine, mucin-conforming amino acid, and increased autophagy-related forkhead-box O signalling. Furthermore, threonine itself affects melanogenesis and metastatic activity in melanoma cells. CONCLUSIONS: We identified a significant association between MUCL1 and threonine with melanogenesis and metastasis-related genes in melanoma cells. Our results define a novel mechanism of mucin regulation, suggesting diagnostic and preventive roles of MUCL1 in cutaneous melanoma.


Assuntos
Melanoma , Neoplasias Cutâneas , Humanos , Melaninas , Melanócitos/metabolismo , Melanoma/patologia , Monofenol Mono-Oxigenase/metabolismo , Mucinas/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/metabolismo , Treonina/metabolismo
14.
Am J Surg Pathol ; 46(5): 637-642, 2022 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-34545857

RESUMO

The distinction between mucinous carcinomas (MCs) and mucocele-like lesions (MLLs), particularly those containing detached epithelial fragments, can be problematic in the limited samples afforded by breast core needle biopsies (CNBs). Neovascularization of mucin has been proposed as a criterion to distinguish MC from MLL, but its value in helping to categorize mucin-producing breast lesions in CNB has not been previously investigated. To address this, we evaluated mucin neovascularization on hematoxylin and eosin (H&E)-stained sections of 140 CNB containing mucin-producing breast lesions including 52 MC, 17 mucin-producing ductal carcinoma in situ (mDCIS), and 71 MLL. In 116 cases with sufficient remaining material (42 MC, 16 mDCIS, and 58 MLL), we also assessed mucin neovascularization on CD31 immunostains. On H&E-stained sections, neovascularization of mucin, defined as delicate, thin-walled microvessels in mucin, and unassociated with fibrous septae, was identified significantly more frequently in MC than in MLL (69.2% vs. 14.1%; P=0.0001). The difference in the frequency of mucin neovascularization between MC and MLL was even greater on CD31 immunostains (97.6% vs. 13.8%, P<0.00001). The sensitivity, specificity, positive predictive value, and negative predictive value of mucin neovascularization for categorizing a lesion as MC were 69.2%, 85.8%, 78.3%, and 79.2%, respectively, for H&E-stained sections and 97.6%, 86.2%, 83.7%, and 98.0%, respectively, for CD31 immunostains. We conclude that mucin neovascularization is significantly more common in MC than in MLL in breast CNB on H&E-stained sections and particularly on CD31 immunostains and may be a valuable adjunct in distinguishing between MC and MLL in problematic cases.


Assuntos
Adenocarcinoma Mucinoso , Neoplasias da Mama , Carcinoma Intraductal não Infiltrante , Mucocele , Adenocarcinoma Mucinoso/diagnóstico , Adenocarcinoma Mucinoso/patologia , Biópsia com Agulha de Grande Calibre , Mama/patologia , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Carcinoma Intraductal não Infiltrante/patologia , Feminino , Humanos , Mucinas , Mucocele/diagnóstico , Mucocele/patologia , Neovascularização Patológica/patologia
15.
Int J Mol Sci ; 23(23)2022 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-36499686

RESUMO

The polypeptide TFF3 belongs to the trefoil factor family (TFF) of lectins. TFF3 is typically secreted from mucous epithelia together with mucins. Both intestinal and salivary TFF3 mainly exist as disulfide-linked heterodimers with IgG Fc binding protein (FCGBP). Here, we investigated bronchial tissue specimens, bronchial secretions, and bronchoalveolar lavage (BAL) fluid from patients with a chronic obstructive pulmonary disease (COPD) background by fast protein liquid chromatography and proteomics. For the first time, we identified different molecular forms of TFF3 in the lung. The high-molecular mass form represents TFF3-FCGBP oligomers, whereas the low-molecular mass forms are homodimeric and monomeric TFF3 with possibly anti-apoptotic activities. In addition, disulfide-linked TFF3 heterodimers with an Mr of about 60k and 30k were detected in both bronchial secretions and BAL fluid. In these liquids, TFF3 is partly N-terminally truncated probably by neutrophil elastase cleavage. TFF3-FCGBP is likely involved in the mucosal innate immune defense against microbial infections. We discuss a hypothetical model how TFF3 might control FCGBP oligomerization. Furthermore, we did not find indications for interactions of TFF3-FCGBP with DMBT1gp340 or the mucin MUC5AC, glycoproteins involved in mucosal innate immunity. Surprisingly, bronchial MUC5AC appeared to be degraded when compared with gastric MUC5AC.


Assuntos
Proteínas de Transporte , Mucinas , Humanos , Brônquios/metabolismo , Moléculas de Adesão Celular/metabolismo , Dissulfetos/metabolismo , Imunoglobulina G/metabolismo , Mucinas/metabolismo , Fator Trefoil-2/metabolismo , Fator Trefoil-3/metabolismo , Fragmentos Fc das Imunoglobulinas
16.
Nat Commun ; 13(1): 7808, 2022 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-36528693

RESUMO

Methods capable of manipulating bacterial colonization are of great significance for modulating host-microbiota relationships. Here, we describe a strategy of in-situ chemical reaction-mediated covalent localization of bacteria. Through a simple one-step imidoester reaction, primary amino groups on bacterial surface can be converted to free thiols under cytocompatible conditions. Surface thiolation is applicable to modify diverse strains and the number of introduced thiols per bacterium can be easily tuned by varying feed ratios. These chemically reactive bacteria are able to spontaneously bond with mucous layer by catalyst-free thiol-disulfide exchange between mucin-associated disulfides and newly converted thiols on bacterial surface and show thiolation level-dependent attachment. Bacteria optimized with 9.3 × 107 thiols per cell achieve 170-fold higher attachment in mucin-enriched jejunum, a challenging location for gut microbiota to colonize. As a proof-of-concept application for microbiota transplantation, covalent bonding-assisted localization of an oral probiotic in the jejunum generates an improved remission of jejunal mucositis. Our findings demonstrate that transforming bacteria with a reactive surface provides an approach to chemically control bacterial localization, which is highly desirable for developing next-generation bacterial living bioagents.


Assuntos
Dissulfetos , Probióticos , Dissulfetos/química , Compostos de Sulfidrila/química , Mucinas , Bactérias
17.
BMC Gastroenterol ; 22(1): 522, 2022 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-36526975

RESUMO

BACKGROUND: Early prediction of severe acute pancreatitis (SAP) plays an important role in timely treatment decisions. Soluble T cell immunoglobulin and mucin domain-3 (sTIM-3) has been applied as a potential biomarker for the prediction of many diseases, while its predictive ability for AP severity remains largely unexplored. In this study, we aimed to identify whether serum sTIM-3 could be used as an indicator of AP severity in the early stage of the disease. METHODS: A retrospective study was conducted. The enrolled AP patients should meet the 2012 Atlanta guideline and have an onset to admission ≤ 48 h. RESULTS: A total of 94 AP patients were enrolled in the current analysis, including 42 (45%), 35 (37%), and 17 (18%) patients were diagnosed as mild AP (MAP), moderately SAP (MSAP), and SAP, respectively. SAP patients had significantly higher the white blood cells (WBCs) count, red blood cells (RBCs) count, C-reactive protein (CRP) level, direct bilirubin level, creatinine and procalcitonin levels compared with MAP and MSAP patients. Among SAP and MSAP patients, significantly higher APACHE II, BISAP, and MCTSI scores were observed compared with MAP patients, and there was significant difference in APACHE II and BISAP scores between SAP and MSAP patients. Stepwise multivariate linear regression analysis showed that the concentrations of serum sTIM-3, as well as the BISAP and MCTSI scores, were significantly associated with the severity of AP. The areas under the ROC curve were 0.914 (95% CI, 0.865-0.963), 0.855 (95%CI, 0.742-0.968) 0.853 (95%CI, 0.768-0.938), and 0.746 (95%CI, 0.633-0.860) for BISAP score, APACHE II score, sTIM-3 level, and MCTSI score, respectively. CONCLUSIONS: Serum sTIM-3 might be ultimately incorporated into a predictive system for assessing the severity of AP.


Assuntos
Pancreatite , Humanos , Pancreatite/diagnóstico , Estudos Retrospectivos , Doença Aguda , Índice de Gravidade de Doença , Prognóstico , Biomarcadores , Imunoglobulinas , Linfócitos T , Mucinas , Valor Preditivo dos Testes
18.
PLoS One ; 17(12): e0276445, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36548335

RESUMO

CRISPR-Cas9-mediated leptin (Lep) knockout (KO) mice exhibited prominent phenotypes for constipation, even though they were not compared with other model animals. This study compared the stool excretion, gastrointestinal motility, histological structure, mucin secretion, and enteric nerve function in Lep KO and high fat diet (HFD)-treated mice to determine if there were differences in their phenotypes for constipation. Most obesity phenotypes, including fat weight, adipocyte size, expression of lipolytic proteins (HSL, perilipin, and ATGL), and glucose concentrations, were detected similarly in the Lep KO and HFD-treated mice. They showed a similar decrease in the excretion parameters, including the stool number, weight, and water content, while the same pattern was detected in the gastrointestinal motility and intestinal length. A similar decrease in the mucosal layer thickness, muscle thickness, ability for mucin secretion, and expression of water channel (aquaporin 3 and 8) genes was detected in the mid-colon of the Lep KO and HFD-treated mice, but the alteration rate in some levels was greater in the HFD-treated group than the Lep KO mice. On the other hand, the levels of c-kit, nNOS, NSE, and PGP9.5 expression for the enteric neurons and intestitial cells of Cajal (ICC) were remarkably lower in the mid-colon of the HFD-treated mice than in the Lep KO mice, but the level of most proteins in both groups remained lower than those in the control group. A similar alteration pattern in the expression of muscarinic acetylcholine receptors (mAChRs) and serotonin receptors was detected in the Lep KO and HFD-treated mice. These results suggest that most phenotypes for obesity-induced constipation were similarly detected in the Lep KO and HFD-treated mice, but there was a difference in the regulatory function of the enteric nervous system (ENS).


Assuntos
Constipação Intestinal , Dieta Hiperlipídica , Leptina , Obesidade , Animais , Camundongos , Constipação Intestinal/etiologia , Constipação Intestinal/genética , Constipação Intestinal/fisiopatologia , Dieta Hiperlipídica/efeitos adversos , Leptina/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mucinas/genética , Obesidade/complicações , Fenótipo
19.
Sci Rep ; 12(1): 21640, 2022 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-36517529

RESUMO

Although probiotics are often indiscriminately prescribed, they are not equal and their effects on the host may profoundly differ. In vitro determination of the attributes of probiotics should be a primary concern and be performed even before clinical studies are designed. In fact, knowledge on the biological properties a microbe possesses is crucial for selecting the most suitable bacteriotherapy for each individual. Herein, nine strains (Bacillus clausii NR, OC, SIN, T, Bacillus coagulans ATCC 7050, Bifidobacterium breve DSM 16604, Limosilactobacillus reuteri DSM 17938, Lacticaseibacillus rhamnosus ATCC 53103, and Saccharomyces boulardii CNCM I-745) declared to be contained in six commercial formulations were tested for their ability to tolerate simulated intestinal conditions, adhere to mucins, and produce ß-galactosidase, antioxidant enzymes, riboflavin, and D-lactate. With the exception of B. breve, all microbes survived in simulated intestinal fluid. L. rhamnosus was unable to adhere to mucins and differences in mucin adhesion were evidenced for L. reuteri and S. boulardii depending on oxygen levels. All microorganisms produced antioxidant enzymes, but only B. clausii, B. coagulans, B. breve, and L. reuteri synthesize ß-galactosidase. Riboflavin secretion was observed for Bacillus species and L. rhamnosus, while D-lactate production was restricted to L. reuteri and L. rhamnosus. Our findings indicate that the analyzed strains possess different in vitro biological properties, thus highlighting the usefulness of in vitro tests as prelude for clinical research.


Assuntos
Probióticos , Antioxidantes , beta-Galactosidase , Mucinas , Lactatos , Riboflavina
20.
BMC Oral Health ; 22(1): 639, 2022 12 24.
Artigo em Inglês | MEDLINE | ID: mdl-36566172

RESUMO

BACKGROUND: Saliva possesses antiviral activity, with submandibular-sublingual (SMSL) saliva having higher antiviral activity than parotid saliva. Various salivary proteins have inactivating effects on influenza A virus (IAV), but the detailed relationship between antiviral proteins and salivary anti-IAV activities in the parotid and SMSL glands is unknown. Here, to identify salivary proteins with anti-IAV activity, salivary proteins from parotid and SMSL glands were identified, quantified, and compared using liquid chromatography-mass spectrometry. METHODS: Twelve healthy male volunteers participated in the study. Parotid and SMSL saliva was collected by suction and collection devices. We assessed anti-IAV activities, protein concentrations, and protein-bound sialic acid concentrations in parotid and SMSL saliva. RESULTS: SMSL had significantly higher anti-IAV activity than parotid saliva. SMSL also had higher concentrations of glycoproteins, such as mucin 5B and mucin 7, protein-bound sialic acid, cystatins, and lysozyme C, compared with parotid saliva. Salivary mucin 5B and mucin 7 concentrations significantly positively correlated with the salivary protein-bound sialic acid concentration. Salivary anti-IAV activity significantly positively correlated with protein-bound sialic acid, mucin 5B, mucin 7, cystatin-C, -S, and -SN concentrations. CONCLUSION: Salivary mucins, cystatins, and lysozyme C contribute to the high anti-IAV activity of SMSL saliva.


Assuntos
Antivirais , Influenzavirus A , Mucina-5B , Saliva , Proteínas e Peptídeos Salivares , Humanos , Masculino , Mucina-5B/análise , Mucina-5B/metabolismo , Mucinas/análise , Mucinas/metabolismo , Muramidase/metabolismo , Ácido N-Acetilneuramínico/análise , Ácido N-Acetilneuramínico/metabolismo , Glândula Parótida , Saliva/química , Proteínas e Peptídeos Salivares/metabolismo , Glândula Submandibular/química , Glândula Submandibular/metabolismo
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