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1.
FASEB J ; 38(13): e23763, 2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-38954404

RESUMO

Riemerella anatipestifer is a pathogenic bacterium that causes duck serositis and meningitis, leading to significant harm to the duck industry. To escape from the host immune system, the meningitis-causing bacteria must survive and multiply in the bloodstream, relying on specific virulence factors such as capsules. Therefore, it is essential to study the genes involved in capsule biosynthesis in R. anatipestifer. In this study, we successfully constructed gene deletion mutants Δ3820 and Δ3830, targeting the GE296_RS03820 and GE296_RS03830 genes, respectively, using the RA-LZ01 strain as the parental strain. The growth kinetics analysis revealed that these two genes contribute to bacterial growth. Transmission and scanning electron microscopy (TEM and SEM) and silver staining showed that Δ3820 and Δ3830 produced the altered capsules and compounds of capsular polysaccharides (CPSs). Serum resistance test showed the mutants also exhibited reduced C3b deposition and decreased resistance serum killing. In vivo, Δ3820 and Δ3830 exhibited markedly declining capacity to cross the blood-brain barrier, compared to RA-LZ01. These findings indicate that the GE296_RS03820 and GE296_RS03830 genes are involved in CPSs biosynthesis and play a key role in the pathogenicity of R. anatipestifer. Furthermore, Δ3820 and Δ3830 mutants presented a tendency toward higher survival rates from RA-LZ01 challenge in vivo. Additionally, sera from ducklings immunized with the mutants showed cross-immunoreactivity with different serotypes of R. anatipestifer, including 1, 2, 7 and 10. Western blot and SDS-PAGE assays revealed that the altered CPSs of Δ3820 and Δ3830 resulted in the exposure of some conserved proteins playing the key role in the cross-immunoreactivity. Our study clearly demonstrated that the GE296_RS03820 and GE296_RS03830 genes are involved in CPS biosynthesis in R. anatipestifer and the capsule is a target for attenuation in vaccine development.


Assuntos
Cápsulas Bacterianas , Patos , Infecções por Flavobacteriaceae , Riemerella , Riemerella/genética , Riemerella/patogenicidade , Riemerella/metabolismo , Animais , Patos/microbiologia , Cápsulas Bacterianas/genética , Cápsulas Bacterianas/metabolismo , Infecções por Flavobacteriaceae/microbiologia , Infecções por Flavobacteriaceae/veterinária , Doenças das Aves Domésticas/microbiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Polissacarídeos Bacterianos/biossíntese , Fatores de Virulência/genética , Deleção de Genes
2.
Anal Chim Acta ; 1316: 342837, 2024 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-38969427

RESUMO

Monitoring the levels of L-Tryptophan (L-Trp) in body fluids is crucial due to its significant role in metabolism and protein synthesis, which ultimately affects neurological health. Herein, we have developed a novel magneto-responsive electrochemical enantioselective sensor for the recognition of L-Trp based on oriented biochar derived from Loofah, Fe3O4 nanoparticles, and molecularly imprinted polydopamine (MIPDA) in xanthan hydrogel. The successful synthesis of these materials has been confirmed through physicochemical and electrochemical characterization. Various operational factors such as pH, response time, loading sample volume, and loading of active materials were optimized. As a result, the sensor exhibited an affordable linear range of 1.0-60.0 µM, with a desirable limit of detection of 0.44 µM. Furthermore, the proposed electrochemical sensor demonstrated good reproducibility and desirable selectivity for the determination of L-Trp, making it suitable for analyzing L-Trp levels in human plasma and serum samples. The development presented offers an appealing, easily accessible, and efficient strategy. It utilizes xanthan hydrogel to improve mass transfer and adhesion, biochar-stabilized Fe3O4 to facilitate magnetic orientation and accelerate mass transfer and sensitivity, and polydopamine MIP to enhance selectivity. This approach enables on-site evaluation of L-Trp levels, which holds significant value for healthcare monitoring and early detection of related conditions.


Assuntos
Técnicas Eletroquímicas , Hidrogéis , Polissacarídeos Bacterianos , Triptofano , Triptofano/química , Triptofano/sangue , Polissacarídeos Bacterianos/química , Hidrogéis/química , Estereoisomerismo , Humanos , Impressão Molecular , Polímeros/química , Polímeros Molecularmente Impressos/química , Indóis/química , Biopolímeros/química , Limite de Detecção , Nanopartículas de Magnetita/química
3.
J Nanobiotechnology ; 22(1): 417, 2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-39014353

RESUMO

Sirolimus (SR) is a macrolide with antifungal and antitumor immunosuppressant properties, classified as a selective inhibitor of mammalian target of rapamycin (mTOR). In this study, an ionic in situ gel of SR (SR-SUS-ISG) was formulated using gellan gum, exhibiting stability regardless of temperature and pH variations, causing minimal irritation. Harnessing the physiological conditions of the eye, SR-SUS-ISG underwent gelation upon contact with ions, increasing drug viscosity and prolonging retention on the ocular surface. Concurrently, SR-SUS-ISG displayed favorable shear dilution properties, reducing viscosity at ambient temperature, enhancing fluidity, and facilitating convenient packaging and transport. Biocompatibility assessments on both human corneal epithelial cells and rabbit eyes demonstrated that SR-SUS-ISG could well be tolerated. Pharmacokinetic investigations in rabbit ocular aqueous humor revealed sustained release, improved corneal penetration, and enhanced bioavailability. Additionally, in a rat corneal alkali burn model, SR-SUS-ISG exhibited inhibitory effects on corneal neovascularization, associated with decreased levels of the inflammatory factors VEGF and MMPs. These findings suggested that SR-SUS-ISG held promise as an effective ocular drug delivery system.


Assuntos
Géis , Sirolimo , Animais , Coelhos , Sirolimo/farmacologia , Sirolimo/farmacocinética , Sirolimo/química , Humanos , Géis/química , Córnea/efeitos dos fármacos , Córnea/metabolismo , Ratos , Masculino , Polissacarídeos Bacterianos/química , Nanopartículas/química , Administração Oftálmica , Neovascularização da Córnea/tratamento farmacológico , Ratos Sprague-Dawley , Viscosidade , Sistemas de Liberação de Medicamentos , Soluções Oftálmicas/química , Soluções Oftálmicas/farmacologia , Linhagem Celular , Disponibilidade Biológica
4.
Curr Microbiol ; 81(8): 260, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38980435

RESUMO

Viral diseases are a serious threat to humans while the most antiviral drugs have low efficiency and side effects on human health. Therefore, using microbial biopolymers as the drugs alternate to treat viral infections seems cost-effective and human friendly option. In the present study, thirty-four exopolysaccharides (EPSs) producing bacteria were isolated, and EPSs production capacity of five salt-tolerant isolates was determined under 0, 100 and 150 mM NaCl. Among these, two isolates exhibiting high anti-coliphage activity were identified through 16S rRNA gene analysis. Moreover, the EPSs were characterized by Fourier-transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis, and their composition was determined. Five salt-tolerant bacteria (MK1, MK2, MK10, MK22 and MK29) exhibited higher production of EPSs at 100 mM NaCl compared to that under non-saline control. At 100 mM NaCl, the yield of EPSs ranged between 105 and 330 mg 100 mL-1 broth. The EPSs produced by the isolates MK1 and MK2 exhibited higher anti-coliphage activity (plaque forming unit decreased from 43 × 106 mL-1 to 3 × 106 and 4 × 106 mL-1, respectively), and were comprised of glucose, fructose, galactose, sucrose, lactose and xylose sugars. FTIR spectroscopy depicted that EPSs are mainly composed of hydroxyl, aliphatic, carboxyl, sulfate and phosphate functional groups, which could have bound coliphage and thus conferred higher anti-coliphage activities to the EPSs. Phylogenetic analysis revealed that MK1 and MK2 isolates formed clades within genus Priestia and Bacillus sequences, respectively. High EPSs production capacity of bacterial isolates under saline condition and high anti-coliphage activity of the EPSs implies that bacterial biopolymers could be useful in antiviral drugs therapy.


Assuntos
Antivirais , Bacillus , Polissacarídeos Bacterianos , RNA Ribossômico 16S , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/metabolismo , Antivirais/farmacologia , Antivirais/química , RNA Ribossômico 16S/genética , Bacillus/genética , Bacillus/metabolismo , Bacillus/química , Bacillus/classificação , Filogenia , Espectroscopia de Infravermelho com Transformada de Fourier , Cloreto de Sódio/farmacologia , Cloreto de Sódio/metabolismo
5.
Int J Mol Sci ; 25(13)2024 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-39000292

RESUMO

Latilactobacillus (L.) sakei is a species of lactic acid bacteria (LAB) mostly studied according to its application in food fermentation. Previously, L. sakei L3 was isolated by our laboratory and possessed the capability of high exopolysaccharide (EPS) yield during sucrose-added fermentation. However, the understanding of sucrose promoting EPS production is still limited. Here, we analyzed the growth characteristics of L. sakei L3 and alterations of its transcriptional profiles during sucrose-added fermentation. The results showed that L. sakei L3 could survive between pH 4.0 and pH 9.0, tolerant to NaCl (<10%, w/v) and urea (<6%, w/v). Meanwhile, transcriptomic analysis showed that a total of 426 differentially expressed genes and eight non-coding RNAs were identified. Genes associated with sucrose metabolism were significantly induced, so L. sakei L3 increased the utilization of sucrose to produce EPS, while genes related to uridine monophosphate (UMP), fatty acids and folate synthetic pathways were significantly inhibited, indicating that L. sakei L3 decreased self-growth, substance and energy metabolism to satisfy EPS production. Overall, transcriptome analysis provided valuable insights into the mechanisms by which L. sakei L3 utilizes sucrose for EPS biosynthesis. The study provided a theoretical foundation for the further application of functional EPS in the food industry.


Assuntos
Fermentação , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Latilactobacillus sakei , Polissacarídeos Bacterianos , Sacarose , Polissacarídeos Bacterianos/biossíntese , Polissacarídeos Bacterianos/metabolismo , Sacarose/metabolismo , Latilactobacillus sakei/metabolismo , Latilactobacillus sakei/genética , Transcriptoma , Concentração de Íons de Hidrogênio
6.
Nutrients ; 16(13)2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38999863

RESUMO

The emptying rate of specific nutrients in enteral formulas is poorly understood, despite the importance of controlling the emptying rate in tube-fed patients. Because of their viscosity, thickened formulas are widely used to avoid gastric reflux and reduce the burden on caregivers. This study examined how thickeners in enteral formulas affected the gastric emptying rates of proteins and carbohydrates. A semi-dynamic gastric model was used to prepare and digest test enteral formulas that contained either no thickeners or agar (0.2%). The amounts of protein and carbohydrates in each emptied aliquot were determined, and the emptying rate was calculated. We found that agar accelerated protein emptying, and an exploratory experiment with agar (0.5%) suggested the possibility of concentration dependence. Additionally, experiments using gellan gum (0.08%), guar gum (0.2%), or carrageenan (0.08%, 0.2%) suggested that protein emptying could vary depending on the thickener type and that carrageenan might slow it. These results could help with the appropriate selection of thickeners added to liquid foods based on the patient's metabolic profile to manage nutrition, not only for tube-fed patients but also for those with oropharyngeal dysphagia or diabetes.


Assuntos
Proteínas Alimentares , Nutrição Enteral , Alimentos Formulados , Galactanos , Esvaziamento Gástrico , Mananas , Gomas Vegetais , Esvaziamento Gástrico/efeitos dos fármacos , Nutrição Enteral/métodos , Humanos , Mananas/farmacologia , Mananas/administração & dosagem , Viscosidade , Galactanos/farmacologia , Proteínas Alimentares/administração & dosagem , Carboidratos da Dieta/administração & dosagem , Carragenina , Ágar , Polissacarídeos Bacterianos/farmacologia , Modelos Biológicos
7.
Microb Genom ; 10(6)2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38896467

RESUMO

Since the introduction of the 13-valent pneumococcal conjugate vaccine (PCV13) in Malawi in 2011, there has been persistent carriage of vaccine serotype (VT) Streptococcus pneumoniae, despite high vaccine coverage. To determine if there has been a genetic change within the VT capsule polysaccharide (cps) loci since the vaccine's introduction, we compared 1022 whole-genome-sequenced VT isolates from 1998 to 2019. We identified the clonal expansion of a multidrug-resistant, penicillin non-susceptible serotype 23F GPSC14-ST2059 lineage, a serotype 14 GPSC9-ST782 lineage and a novel serotype 14 sequence type GPSC9-ST18728 lineage. Serotype 23F GPSC14-ST2059 had an I253T mutation within the capsule oligosaccharide repeat unit polymerase Wzy protein, which is predicted in silico to alter the protein pocket cavity. Moreover, serotype 23F GPSC14-ST2059 had SNPs in the DNA binding sites for the cps transcriptional repressors CspR and SpxR. Serotype 14 GPSC9-ST782 harbours a non-truncated version of the large repetitive protein (Lrp), containing a Cna protein B-type domain which is also present in proteins associated with infection and colonisation. These emergent lineages also harboured genes associated with antibiotic resistance, and the promotion of colonisation and infection which were absent in other lineages of the same serotype. Together these data suggest that in addition to serotype replacement, modifications of the capsule locus associated with changes in virulence factor expression and antibiotic resistance may promote vaccine escape. In summary, the study highlights that the persistence of vaccine serotype carriage despite high vaccine coverage in Malawi may be partly caused by expansion of VT lineages post-PCV13 rollout.


Assuntos
Cápsulas Bacterianas , Infecções Pneumocócicas , Vacinas Pneumocócicas , Sorogrupo , Streptococcus pneumoniae , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/patogenicidade , Vacinas Pneumocócicas/imunologia , Humanos , Malaui , Cápsulas Bacterianas/genética , Infecções Pneumocócicas/microbiologia , Infecções Pneumocócicas/prevenção & controle , Vacinas Conjugadas , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/imunologia , Virulência/genética , Genótipo , Sequenciamento Completo do Genoma , Proteínas de Bactérias/genética , Fatores de Virulência/genética , Pré-Escolar , Polimorfismo de Nucleotídeo Único , Lactente , Masculino
8.
Pol J Microbiol ; 73(2): 237-252, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38905279

RESUMO

This study aimed to elucidate the influence of various culture medium components, including carbon sources, nitrogen sources, inorganic salts, suspension agents, and temperature, on the mycelial growth characteristics of Phallus dongsun. Employing single-factor experiments and response surface methodology within glass Petri dishes, the research identified that carrot powder, soybean powder, and ZnSO4 notably enhanced the proliferation of aerial mycelium, significantly augmenting the growth rate of P. dongsun mycelium. The resultant mycelium was observed to be dense, robust, and fluffy in texture. In particular, ZnSO4 markedly accelerated the mycelium growth rate. Furthermore, xanthan gum was found to effectively modulate the medium's viscosity, ensuring a stable suspension and facilitating nutrient equilibrium. The optimal cultivation temperature was determined to be 25°C, with mycelial growth ceasing below 5°C and mycelium perishing at temperatures exceeding 35°C. The optimal medium composition was established as follows: wheat starch 5 g/l, carrot powder 5 g/l, soybean powder 7.50 g/l, glucose 10 g/l, ZnSO4 0.71 g/l, NH4Cl 0.68 g/l, xanthan gum 0.5 g/l, and agar 20 g/l. Under these optimized conditions, the mycelium of P. dongsun exhibited a rapid growth rate (1.04 ± 0.14 mm/day), characterized by a thick, dense, and well-developed structure. This investigation provides a theoretical foundation for the conservation, strain selection, and breeding of P. dongsun.


Assuntos
Meios de Cultura , Micélio , Temperatura , Micélio/crescimento & desenvolvimento , Meios de Cultura/química , Nitrogênio , Carbono/química , Polissacarídeos Bacterianos/química
9.
Molecules ; 29(11)2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38893550

RESUMO

Transparent soil (TS) presents immense potential for root phenotyping due to its ability to facilitate high-resolution imaging. However, challenges related to transparency, mechanical properties, and cost hinder its development. Herein, we introduce super-transparent soil (s-TS) prepared via the droplet method using low acyl gellan gum and hydroxyethyl cellulose crosslinked with magnesium ions. The refractive index of the hydroxyethyl cellulose solution (1.345) closely aligns with that of water (1.333) and the low acyl gellan gum solution (1.340), thereby significantly enhancing the transmittance of hydrogel-based transparent soil. Optimal transmittance (98.45%) is achieved with polymer concentrations ranging from 0.8 to 1.6 wt.% and ion concentrations between 0.01 and 0.09 mol·L-1. After 60 days of plant cultivation, s-TS maintains a transmittance exceeding 89.5%, enabling the detailed visualization of root growth dynamics. Furthermore, s-TS exhibits remarkable mechanical properties, withstanding a maximum compressive stress of 477 kPa and supporting a maximum load-bearing depth of 186 cm. This innovative approach holds promising implications for advanced root phenotyping studies, fostering the investigation of root heterogeneity and the development of selective expression under controlled conditions.


Assuntos
Fenótipo , Raízes de Plantas , Solo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/química , Solo/química , Polissacarídeos Bacterianos/química
10.
Front Immunol ; 15: 1388721, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38840926

RESUMO

The disaccharide (ß-D-glucopyranosyluronic acid)-(1→4)-ß-D-glucopyranoside represents a repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 3. A conjugate of the disaccharide with BSA (di-BSA conjugate) adjuvanted with aluminum hydroxide induced - in contrast to the non-adjuvanted conjugate - IgG1 antibody production and protected mice against S. pneumoniae serotype 3 infection after intraperitoneal prime-boost immunization. Adjuvanted and non-adjuvanted conjugates induced production of Th1 (IFNγ, TNFα); Th2 (IL-5, IL-13); Th17 (IL-17A), Th1/Th17 (IL-22), and Th2/Th17 cytokines (IL-21) after immunization. The concentration of cytokines in mice sera was higher in response to the adjuvanted conjugate, with the highest level of IL-17A production after the prime and boost immunizations. In contrast, the non-adjuvanted conjugate elicited only weak production of IL-17A, which gradually decreased after the second immunization. After boost immunization of mice with the adjuvanted di-BSA conjugate, there was a significant increase in the number of CD45+/CD19+ B cells, TCR+ γδ T cell, CD5+ В1 cells, and activated cells with MHC II+ expression in the spleens of the mice. IL-17A, TCR+ γδ T cells, and CD5+ В1 cells play a crucial role in preventing pneumococcal infection, but can also contribute to autoimmune diseases. Immunization with the adjuvanted and non-adjuvanted di-BSA conjugate did not elicit autoantibodies against double-stranded DNA targeting cell nuclei in mice. Thus, the molecular and cellular markers associated with antibody production and protective activity in response to immunization with the di-BSA conjugate adjuvanted with aluminum hydroxide are IL-17A, TCR+ γδ T cells, and CD5+ В1 cells against the background of increasing MHC II+ expression.


Assuntos
Interleucina-17 , Vacinas Pneumocócicas , Soroalbumina Bovina , Streptococcus pneumoniae , Animais , Interleucina-17/imunologia , Interleucina-17/metabolismo , Streptococcus pneumoniae/imunologia , Camundongos , Soroalbumina Bovina/imunologia , Vacinas Pneumocócicas/imunologia , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/prevenção & controle , Dissacarídeos/imunologia , Cápsulas Bacterianas/imunologia , Polissacarídeos Bacterianos/imunologia , Adjuvantes Imunológicos/administração & dosagem , Feminino , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Linfócitos Intraepiteliais/imunologia , Sorogrupo , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo
11.
Int J Biol Macromol ; 273(Pt 2): 133091, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38878924

RESUMO

The increasing significance of biopolymer-based food packaging can be attributed to its biodegradability and independence from petroleum-derived materials. Concurrently, metal oxide nanoparticles (NPs) have gained prominence as effective antimicrobial agents against both wild-type and antibiotic-resistant microbes. In this study, cerium oxide or ceria, CeO2, nanoparticles with an average diameter of 50 nm were synthesized via a green method utilizing Vibrio sp. VLC cell lysate supernatant. The synthesized CeO2 NPs displayed remarkable antimicrobial properties, inhibiting the growth of Escherichia coli and Staphylococcus aureus by 93.7 % and 98 %, respectively. To enhance the potential of bacterial cellulose (BC) for advanced applications, we developed a BC/xanthan/CeO2 nanocomposite using both ex situ and in situ techniques. The integration of CeO2 NPs within the nanocomposite structure not only improved the inherent properties of BC, but also rendered it suitable for use in active food packaging systems. The nanocomposite exhibited no significant cytotoxicity on the human dermal fibroblast (HDF) cells, confirming its safety. Nanocomposites containing biogenically synthesized CeO2 NPs demonstrated exceptional efficacy for reducing microbial contamination. Bread samples coated with nanocomposite films displayed no signs of microbial growth. These results support the application of BC/xanthan/CeO2 nanocomposites as suitable and effective coating materials for antimicrobial food packaging applications.


Assuntos
Antibacterianos , Celulose , Cério , Embalagem de Alimentos , Nanocompostos , Polissacarídeos Bacterianos , Celulose/química , Celulose/farmacologia , Embalagem de Alimentos/métodos , Cério/química , Cério/farmacologia , Nanocompostos/química , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia , Antibacterianos/farmacologia , Antibacterianos/química , Humanos , Staphylococcus aureus/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Nanopartículas Metálicas/química
12.
Int J Biol Macromol ; 273(Pt 2): 133175, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38889835

RESUMO

The molecular and colloidal-level interactions between two major phenolic acids, gallic and caffeic acid, with a major food polysaccharide, xanthan gum, were studied in binary systems aiming to correlate the stability of the binary systems as a function of pH and xanthan-polyphenol concentrations. Global stability diagrams were built, acting as roadmaps for examining the phase separation regimes followed by the fluorimetry-based thermodynamics of the interactions. The effects of noncovalent interactions on the macroscopic behavior of the binary systems were studied, using shear and extensional rheometry. The collected data for caffeic acid - xanthan gum mixtures showed that the main interactions were pH-independent volume exclusions, while gallic acid interacts with xanthan gum, especially at pH 7 with other mechanisms as well, improving the colloidal dispersion stability. A combination of fluorimetry, extensional rheology and stability measurements highlight the effect of gallic acid-induced aggregation of xanthan gum, both in structuring and de-structuring the binary systems. The above provide a coherent framework of the physicochemical aspect of binary systems, shedding light on the role of xanthan gum in its oral functions, such as in inducing texture, in model complex systems containing phenolic acids.


Assuntos
Polissacarídeos Bacterianos , Reologia , Polissacarídeos Bacterianos/química , Hidroxibenzoatos/química , Concentração de Íons de Hidrogênio , Ácido Gálico/química , Termodinâmica
13.
Int J Biol Macromol ; 273(Pt 2): 133215, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38897515

RESUMO

Lactic acid bacteria (LAB) produce a broad spectrum of exopolysaccharides (EPSs), commonly used as texturizers in food products. Due to their potential contribution to LAB probiotic properties, like adhesion to human epithelial cells and competitive exclusion of pathogens from human intestinal epithelial cells, this study was focussed on the structural and functional characterization of the EPSs produced by two Limosilactobacillus fermentum strains - MC1, originating from mother's milk, and D12, autochthonous from Croatian smoked fresh cheese. Whole-genome sequencing and functional annotation of both L. fermentum strains by RAST server revealed the genes involved in EPS production and transport, with some differences in functionally related genes. EPSs were extracted from the cell surface of both bacterial strains and purified by size-exclusion chromatography. Structural characterization of the EPSs, achieved by chemical analyses and 1D and 2D NMR spectroscopy, showed that both strains produce an identical mixture of three different EPSs containing galactofuranose and glucopyranose residues. However, a comparison of the functional properties showed that the MC1 strain adhered better to the Caco-2 cell line and exhibited stronger antimicrobial effect against Salmonella enterica serovar Typhimurium FP1 than the D12 strain, which may be attributed to the potential bacteriocin activity of the MC1 strain.


Assuntos
Limosilactobacillus fermentum , Polissacarídeos Bacterianos , Humanos , Limosilactobacillus fermentum/metabolismo , Limosilactobacillus fermentum/química , Células CACO-2 , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos
14.
Arch Microbiol ; 206(7): 289, 2024 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-38847838

RESUMO

Staphylococcus epidermidis is an opportunistic pathogen commonly implicated in medical device-related infections. Its propensity to form biofilms not only leads to chronic infections but also exacerbates the issue of antibiotic resistance, necessitating high-dose antimicrobial treatments. In this study, we explored the use of diclofenac sodium, a non-steroidal anti-inflammatory drug, as an anti-biofilm agent against S. epidermidis. In this study, crystal violet staining and confocal laser scanning microscope analysis showed that diclofenac sodium, at subinhibitory concentration (0.4 mM), significantly inhibited biofilm formation in both methicillin-susceptible and methicillin-resistant S. epidermidis isolates. MTT assays demonstrated that 0.4 mM diclofenac sodium reduced the metabolic activity of biofilms by 25.21-49.01% compared to untreated controls. Additionally, the treatment of diclofenac sodium resulted in a significant decrease (56.01-65.67%) in initial bacterial adhesion, a crucial early phase of biofilm development. Notably, diclofenac sodium decreased the production of polysaccharide intercellular adhesin (PIA), a key component of the S. epidermidis biofilm matrix, in a dose-dependent manner. Real-time quantitative PCR analysis revealed that diclofenac sodium treatment downregulated biofilm-associated genes icaA, fnbA, and sigB and upregulated negative regulatory genes icaR and luxS, providing potential mechanistic insights. These findings indicate that diclofenac sodium inhibits S. epidermidis biofilm formation by affecting initial bacterial adhesion and the PIA synthesis. This underscores the potential of diclofenac sodium as a supplementary antimicrobial agent in combating staphylococcal biofilm-associated infections.


Assuntos
Antibacterianos , Biofilmes , Diclofenaco , Staphylococcus epidermidis , Biofilmes/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/fisiologia , Diclofenaco/farmacologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Anti-Inflamatórios não Esteroides/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Humanos , Polissacarídeos Bacterianos/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos
15.
Carbohydr Polym ; 339: 122202, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38823898

RESUMO

Interactions among multi-component play a critical role in modulating the foaming properties of aerated foods. This study evaluated the mechanisms of synergistic improvement of gellan gum (GEG) and guar gum (GUG) on the foaming properties of soy protein isolate (SPI)-based complex. The results showed that the GEG/GUG ratio was closely related to the intermolecular interactions of SPI-based ternary complex and the dynamical changing of its foaming properties. The SPI/GEG/GUG ternary complex with a GEG/GUG ratio of 2/3 exhibited the highest foamability (195 %) and comparable foam stability (99.17 %), which were 32.95 % and 2.99 % higher than that of SPI/GEG binary complex. At this ratio, GUG promoted the interactions between SPI and GEG, and bound to complex's surface through hydrogen bonding, resulting in the increase of particle size and surface charge, and the decrease of surface hydrophobicity. Although this reduced the diffusion of complex onto the air/water interface, it increased permeation rate and molecular rearrangement behavior, which were the potential mechanisms to improve the foaming properties. Additionally, the synergistic effect of GEG and GUG also enhanced the elastic strength and solid characteristics of foam systems. This study provided a theoretical guidance for the targeted modulation of foaming properties of multi-component aerated foods.


Assuntos
Galactanos , Mananas , Gomas Vegetais , Polissacarídeos Bacterianos , Proteínas de Soja , Gomas Vegetais/química , Galactanos/química , Proteínas de Soja/química , Mananas/química , Polissacarídeos Bacterianos/química , Interações Hidrofóbicas e Hidrofílicas , Tamanho da Partícula , Ligação de Hidrogênio
16.
Carbohydr Polym ; 339: 122292, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38823937

RESUMO

Through adaptive laboratory evolution (ALE) of Sphingomonas sp. ATCC 31555, fermentation for production of low-molecular-weight welan gum (LMW-WG) was performed using glycerol as sole carbon source. During ALE, GPC-MALS analysis revealed a gradual decrease in WG molecular weight with the increase of adaptation cycles, accompanied by changes in solution conformation. LMW-WG was purified and structurally analyzed using GPC-MALS, monosaccharide composition analysis, infrared spectroscopy, NMR analysis, atomic force microscopy, and scanning electron microscopy. Subsequently, LMW-WG obtains hydration, transparency, antioxidant activity, and rheological properties. Finally, an in vitro simulation colon reactor was used to evaluate potential prebiotic properties of LMW-WG as dietary fiber. Compared with WG produced using sucrose as substrate, LMW-WG exhibited a fourfold reduction in molecular weight while maintaining moderate viscosity. Structurally, L-Rha nearly completely replaced L-Man. Furthermore, LMW-WG demonstrated excellent hydration, antioxidant activity, and high transparency. It also exhibited resistance to saliva and gastrointestinal digestion, showcasing a favorable colonization effect on Bifidobacterium, making it a promising symbiotic agent.


Assuntos
Antioxidantes , Fermentação , Glicerol , Peso Molecular , Sphingomonas , Glicerol/química , Glicerol/metabolismo , Antioxidantes/química , Antioxidantes/farmacologia , Sphingomonas/metabolismo , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia , Viscosidade , Prebióticos , Bifidobacterium/metabolismo
17.
Science ; 384(6701): eado0713, 2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38870284

RESUMO

Bacteria can repurpose their own bacteriophage viruses (phage) to kill competing bacteria. Phage-derived elements are frequently strain specific in their killing activity, although there is limited evidence that this specificity drives bacterial population dynamics. Here, we identified intact phage and their derived elements in a metapopulation of wild plant-associated Pseudomonas genomes. We discovered that the most abundant viral cluster encodes a phage remnant resembling a phage tail called a tailocin, which bacteria have co-opted to kill bacterial competitors. Each pathogenic Pseudomonas strain carries one of a few distinct tailocin variants that target the variable polysaccharides in the outer membrane of co-occurring pathogenic Pseudomonas strains. Analysis of herbarium samples from the past 170 years revealed that the same tailocin and bacterial receptor variants have persisted in Pseudomonas populations. These results suggest that tailocin genetic diversity can be mined to develop targeted "tailocin cocktails" for microbial control.


Assuntos
Bacteriocinas , Fagos de Pseudomonas , Pseudomonas , Proteínas da Cauda Viral , Antibiose , Membrana Externa Bacteriana/metabolismo , Bacteriocinas/genética , Bacteriocinas/metabolismo , Variação Genética , Genoma Bacteriano , Polissacarídeos Bacterianos/metabolismo , Pseudomonas/metabolismo , Pseudomonas/virologia , Fagos de Pseudomonas/genética , Fagos de Pseudomonas/metabolismo , Proteínas da Cauda Viral/metabolismo , Proteínas da Cauda Viral/genética , Terapia por Fagos/métodos
18.
Vet Res ; 55(1): 80, 2024 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-38886823

RESUMO

Bacteria utilize intercellular communication to orchestrate essential cellular processes, adapt to environmental changes, develop antibiotic tolerance, and enhance virulence. This communication, known as quorum sensing (QS), is mediated by the exchange of small signalling molecules called autoinducers. AI-2 QS, regulated by the metabolic enzyme LuxS (S-ribosylhomocysteine lyase), acts as a universal intercellular communication mechanism across gram-positive and gram-negative bacteria and is crucial for diverse bacterial processes. In this study, we demonstrated that in Streptococcus suis (S. suis), a notable zoonotic pathogen, AI-2 QS enhances galactose utilization, upregulates the Leloir pathway for capsular polysaccharide (CPS) precursor production, and boosts CPS synthesis, leading to increased resistance to macrophage phagocytosis. Additionally, our molecular docking and dynamics simulations suggest that, similar to S. pneumoniae, FruA, a fructose-specific phosphoenolpyruvate phosphotransferase system prevalent in gram-positive pathogens, may also function as an AI-2 membrane surface receptor in S. suis. In conclusion, our study demonstrated the significance of AI-2 in the synthesis of galactose metabolism-dependent CPS in S. suis. Additionally, we conducted a preliminary analysis of the potential role of FruA as a membrane surface receptor for S. suis AI-2.


Assuntos
Galactose , Percepção de Quorum , Streptococcus suis , Streptococcus suis/fisiologia , Galactose/metabolismo , Percepção de Quorum/fisiologia , Virulência , Animais , Cápsulas Bacterianas/metabolismo , Lactonas/metabolismo , Infecções Estreptocócicas/veterinária , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/imunologia , Homosserina/análogos & derivados , Homosserina/metabolismo , Polissacarídeos Bacterianos/metabolismo
19.
Nat Commun ; 15(1): 5258, 2024 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-38898034

RESUMO

Many bacterial pathogens, including the human exclusive pathogen Salmonella Typhi, express capsular polysaccharides as a crucial virulence factor. Here, through S. Typhi whole genome sequence analyses and functional studies, we found a list of single point mutations that make S. Typhi hypervirulent. We discovered a single point mutation in the Vi biosynthesis enzymes that control Vi polymerization or acetylation is enough to result in different capsule variants of S. Typhi. All variant strains are pathogenic, but the hyper Vi capsule variants are particularly hypervirulent, as demonstrated by the high morbidity and mortality rates observed in infected mice. The hypo Vi capsule variants have primarily been identified in Africa, whereas the hyper Vi capsule variants are distributed worldwide. Collectively, these studies increase awareness about the existence of different capsule variants of S. Typhi, establish a solid foundation for numerous future studies on S. Typhi capsule variants, and offer valuable insights into strategies to combat capsulated bacteria.


Assuntos
Cápsulas Bacterianas , Mutação de Sentido Incorreto , Polissacarídeos Bacterianos , Salmonella typhi , Febre Tifoide , Salmonella typhi/genética , Salmonella typhi/patogenicidade , Animais , Camundongos , Virulência/genética , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/biossíntese , Polissacarídeos Bacterianos/metabolismo , Cápsulas Bacterianas/genética , Cápsulas Bacterianas/metabolismo , Febre Tifoide/microbiologia , Humanos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Feminino , Sequenciamento Completo do Genoma
20.
Int J Biol Macromol ; 272(Pt 1): 132906, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38851991

RESUMO

Sourdough bread enriched with soluble fiber (by in-situ exopolysaccharides production) and insoluble fiber (by gazpacho by-products addition) showed prebiotic effects an in vitro dynamic colonic fermentation performance with obese volunteer's microbiota. Bifidobacterium population was maintained whereas Lactobacillus increased throughout the colonic sections. Conversely, Enterobacteriaceae and Clostridium groups clearly decreased. Specific bacteria associated with beneficial effects increased in the ascending colon (Lactobacillus fermentum, Lactobacillus paracasei, Bifidobacterium longum and Bifidobacterium adolescentis) whereas Eubacterium eligens, Alistipes senegalensis, Prevotella copri and Eubacterium desmolans increased in the transversal and descending colon. Additionally, Blautia faecis and Ruminococcus albus increased in the transversal colon, and Bifidobacterium longum, Roseburia faecis and Victivallis vadensis in the descending colon. Bifidobacterium and Lactobacillus fermented the in-situ exopolysaccharides and released pectins from gazpacho by-products, as well as cellulosic degraded bacteria. This increased the short and medium chain fatty acids. Acetic acid, as well as butyric acid, increased throughout the colonic tract, which showed greater increases only in the transversal and descending colonic segments. Conversely, propionic acid was slightly affected by the colonic fermentation. These results show that sourdough bread is a useful food matrix for the enrichment of vegetable by-products (or other fibers) in order to formulate products with microbiota modulatory capacities.


Assuntos
Pão , Disbiose , Fermentação , Pão/microbiologia , Humanos , Disbiose/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Fibras na Dieta/metabolismo , Polissacarídeos Bacterianos/farmacologia , Colo/microbiologia , Colo/metabolismo , Bifidobacterium/metabolismo , Masculino , Lactobacillus/metabolismo
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