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1.
Carbohydr Polym ; 335: 122076, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38616075

RESUMO

The development of exopolysaccharide-based polymers is gaining increasing attention in various industrial biotechnology fields for materials such as thickeners, texture modifiers, anti-freeze agents, antioxidants, and antibacterial agents. High-viscosity carboxyethyl-succinoglycan (CE-SG) was directly synthesized from succinoglycan (SG) isolated from Sinorhizobium meliloti Rm 1021, and its structural, rheological, and physiological properties were investigated. The viscosity of CE-SG gradually increased in proportion to the degree of carboxyethylation substitution. In particular, when the molar ratio of SG and 3-chloropropionic acid was 1:100, the viscosity was significantly improved by 21.18 times at a shear rate of 10 s-1. Increased carboxyethylation of SG also improved the thermal stability of CE-SG. Furthermore, the CE-SG solution showed 90.18 and 91.78 % antibacterial effects against Escherichia coli and Staphylococcus aureus and effective antioxidant activity against DPPH and hydroxyl radicals. In particular, CE-SG hydrogels coordinated with Fe3+ ions, which improved both viscosity and rheological properties, while also exhibiting reduction-responsive drug release through 1,4-dithiothreitol. The results of this study suggest that SG derivatives, such as CE-SG, can be used as functional biomaterials in various fields such as food, cosmetics, and pharmaceutical industries.


Assuntos
Antioxidantes , Hidrogéis , Polissacarídeos Bacterianos , Hidrogéis/farmacologia , Antioxidantes/farmacologia , Antibacterianos/farmacologia , Indústria Farmacêutica , Escherichia coli
2.
Chem Pharm Bull (Tokyo) ; 72(4): 381-384, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38616116

RESUMO

Bietti's crystalline dystrophy (BCD) is an autosomal recessive chorioretinal degeneration caused by mutations in the CYP4V2 gene. It is characterized by cholesterol accumulation and crystal-like deposits in the retinas. Hydroxypropyl-ß-cyclodextrin (HP-ß-CyD) exerts therapeutic effects against BCD by reducing lysosomal dysfunction and inhibiting cytotoxicity in induced pluripotent stem cell (iPSC)-RPE cells established from patient-derived iPS cells. However, the ocular retention of HP-ß-CyD is low and needs to be improved. Therefore, this study used a viscous agent to develop a sustained-release ophthalmic formulation containing HP-ß-CyD. Our results suggest that HP-ß-CyD-containing xanthan gum has a considerably higher sustained release capacity than other viscous agents, such as methylcellulose and sodium alginate. In addition, the HP-ß-CyD-containing xanthan gum exhibited pseudoplastic behavior. It was less cytotoxic to human retinal pigment epithelial cells compared with HP-ß-CyD alone. Furthermore, the slow release of HP-ß-CyD from xanthan gum caused a sustained decrease in free intracellular cholesterol. These results suggest that xanthan gum is a useful substrate for the sustained release formulation of HP-ß-CyD, and that HP-ß-CyD-containing xanthan gum has potential as an eye drop for BCD treatment.


Assuntos
Colesterol , Distrofias Hereditárias da Córnea , Polissacarídeos Bacterianos , Doenças Retinianas , Humanos , Preparações de Ação Retardada/farmacologia , 2-Hidroxipropil-beta-Ciclodextrina/farmacologia
3.
Int J Biol Macromol ; 265(Pt 2): 131159, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38565484

RESUMO

A goal of this study is to explore the difference in rheological properties of xanthan gum (XG)-based mixtures with gum arabic (GA) or GA-based emulsion (GAE). The stability of GAE was improved by thickening with XG. The intrinsic viscosity of all mixtures decreased as GA concentration increased, implying an XG conformational transition from the disordered to the ordered form. All mixtures except for an XG-GA mixture at 6.0 % GA attained a higher consistency index value than XG alone, indicating synergistic interactions between the components. At a high GA concentration (>3.0 %), the XG-GAE mixture showed higher relative apparent viscosity values than the XG-GA mixture. All mixtures except for an XG-GA mixture at 6.0 % GA showed higher elastic modulus and lower viscous modulus values than XG alone. Consequently, all mixtures showed lower tan δ values (0.26-0.30) than XG alone (0.31). Moreover, with a high GA concentration (>1.5 %), the XG-GAE mixtures achieved lower relative tan δ values than XG-GA mixture. These results indicate that XG formed a higher weak gel-like network with GAE than GA. Overall, the findings demonstrate that the interaction between XG and GA is influenced by conformational changes in the latter in both aqueous and emulsion systems.


Assuntos
Goma Arábica , Gomas Vegetais , Emulsões , Polissacarídeos Bacterianos , Viscosidade , Reologia/métodos
4.
Biofabrication ; 16(3)2024 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-38565131

RESUMO

Extrusion-based bioprinting is a promising technology for the fabrication of complex three-dimensional (3D) tissue-engineered constructs. To further improve the printing accuracy and provide mechanical support during the printing process, hydrogel-based support bath materials have been developed. However, the gel structure of some support bath materials can be compromised when exposed to certain bioink crosslinking cues, hence their compatibility with bioinks can be limited. In this study, a xanthan gum-based composite support material compatible with multiple crosslinking mechanisms is developed. Different support bath materials can have different underlying polymeric structures, for example, particulate suspensions and polymer solution with varying supramolecular structure) and these properties are governed by a variety of different intermolecular interactions. However, common rheological behavior can be expected because they have similar demonstrated performance and functionality. To provide a detailed exploration/identification of the common rheological properties expressed by different support bath materials from a unified perspective, benchmark support bath materials from previous studies were prepared. A comparative rheological study revealed both the structural and shear behavior characteristics shared by support bath materials, including yield stress, gel complex moduli, shear-thinning behavior, and self-healing properties. Gel structural stability and functionality of support materials were tested in the presence of various crosslinking stimuli, confirming the versatility of the xanthan-based support material. We further investigated the effect of support materials and the diameter of extrusion needles on the printability of bioinks to demonstrate the improvement in bioink printability and structural integrity. Cytotoxicity and cell encapsulation viability tests were carried out to confirm the cell compatibility of the xanthan gum-based support bath material. We propose and demonstrate the versatility and compatibility of the novel support bath material and provide detailed new insight into the essential properties and behavior of these materials that serve as a guide for further development of support bath-based 3D bioprinting.


Assuntos
Bioimpressão , Engenharia Tecidual , Polissacarídeos Bacterianos , Reologia , Impressão Tridimensional , Bioimpressão/métodos , Hidrogéis/química , Tecidos Suporte/química
5.
Carbohydr Res ; 538: 109075, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38564901

RESUMO

The global demand for therapeutic prebiotics persuades the quest for novel exopolysaccharides that can retard the growth of pathobionts and healthcare-associated pathogens. In this regard, an exopolysaccharide (3.69 mg/mL) producing strain showing prebiotic and antibiofilm activity was isolated from indigenous pineapple pomace of Tripura and identified as Bacillus subtilis PR-C18. Zymogram analysis revealed EPS PR-C18 was synthesized by levansucrase (∼57 kDa) with a maximal activity of 4.62 U/mg. Chromatography techniques, FTIR, and NMR spectral data revealed the homopolymeric nature of purified EPS with a molecular weight of 3.40 × 104 Da. SEM and rheological study unveiled its microporous structure and shear-thinning effect. Furthermore, EPS PR-C18 showed remarkable emulsification, flocculation, water retention, water solubilization, and antioxidant activity. DSC-TGA data demonstrated its high thermostability and cytotoxicity analysis verified its nontoxic biocompatible nature. In addition, the antibiofilm activity of EPS PR-C18 was validated using molecular docking, molecular simulation, MM-GBSA and PCA studies, which exhibited its strong binding affinity (-20.79 kcal/moL) with PelD, a virulence factor from Pseudomonas aeruginosa. Together, these findings support the future exploitation of EPS PR-C18 as an additive or adjuvant in food and pharmaceutical sectors.


Assuntos
Bacillus subtilis , Prebióticos , Simulação de Acoplamento Molecular , Frutanos/farmacologia , Frutanos/química , Biofilmes , Água , Polissacarídeos Bacterianos/farmacologia , Polissacarídeos Bacterianos/química
6.
Int. microbiol ; 27(2): 459-476, Abr. 2024. graf
Artigo em Inglês | IBECS | ID: ibc-232293

RESUMO

Gellan gum (GG) has gained tremendous attention owing to its diversified applications. However, its high production and hence market cost are still a bottleneck in its widespread utilization. In the present study, high GG producing mutant of Sphingomonas spp. was developed by random mutagenesis using ethyl methylsulphonate (EMS) for industrial fermentation and identified as Sphingomonas trueperi after 16S rRNA and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF–MS) analysis. The fermentation conditions such as pH, temperature, and inoculum ratio were optimized by one factor at a time (OFAT) followed by screening of medium components by the Plackett–Burman statistical design. The most critical nutrients were further optimized by response surface methodology for maximizing GG production. The effect of dissolved oxygen tension in bioreactor on cell growth, substrate consumption, GG production, and batch productivity was elucidated. The highest GG titer (23 ± 2.4 g/L) was attained in optimized medium at 10% inoculum (6.45 ± 0.5 log cfu/mL) under controlled fermentation conditions of pH (7), temperature (30 °C), agitation (300–600 rpm), and aeration (0.5–2.0 SLPM) at 22 ± 2% dissolved oxygen tension in a 10-L bioreactor. Kinetic modeling of optimized batch process revealed that logistic growth model could best explain biomass accumulation, while GG formation and substrate consumption were best explained by Luedeking-Piret and exponential decay model, respectively. Structural and physico-functional features of GG produced by mutant Sphingomonas spp. were characterized by HPLC, FTIR, NMR, DSC, TGA, GPC, SEM, and rheological analysis. The higher productivity (0.51 g/L/h) under optimized fermentation conditions suggests potential consideration of mutant and process for commercial utilization.(AU)


Assuntos
Humanos , Mutagênese , Sphingomonas , RNA Ribossômico 16S , Oxigênio , Fermentação , Polissacarídeos Bacterianos
7.
Int J Biol Macromol ; 265(Pt 2): 130747, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38479657

RESUMO

Exopolysaccharides (EPSs), originating from various microbes, and mushrooms, excel in their conventional role in bioremediation to showcase diverse applications emphasizing nanobiotechnology including nano-drug carriers, nano-excipients, medication and/or cell encapsulation, gene delivery, tissue engineering, diagnostics, and associated treatments. Acknowledged for contributions to adsorption, nutrition, and biomedicine, EPSs are emerging as appealing alternatives to traditional polymers, for biodegradability and biocompatibility. This article shifts away from the conventional utility to delve deeply into the expansive landscape of EPS applications, particularly highlighting their integration into cutting-edge nanobiotechnological methods. Exploring EPS synthesis, extraction, composition, and properties, the discussion emphasizes their structural diversity with molecular weight and heteropolymer compositions. Their role as raw materials for value-added products takes center stage, with critical insights into recent applications in nanobiotechnology. The multifaceted potential, biological relevance, and commercial applicability of EPSs in contemporary research and industry align with the nanotechnological advancements coupled with biotechnological nano-cleansing agents are highlighted. EPS-based nanostructures for biological applications have a bright future ahead of them. Providing crucial information for present and future practices, this review sheds light on how eco-friendly EPSs derived from microbial biomass of terrestrial and aquatic environments can be used to better understand contemporary nanobiotechnology for the benefit of society.


Assuntos
Nanoestruturas , Polissacarídeos Bacterianos , Polissacarídeos Bacterianos/química , Biotecnologia , Portadores de Fármacos , Nanotecnologia
8.
Int J Biol Macromol ; 265(Pt 2): 131143, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38537861

RESUMO

Hydrogel electrolyte is an ideal candidate material for flexible energy storage devices due to its excellent softness and conductivity properties. However, challenges such as the inherent mechanical weakness, the susceptibility to be frozen in low-temperature environments, and the insufficiency of hydrogel-electrode contact persist. Herein, a "Multi in One" strategy is employed to effectively conquer these difficulties by endowing hydrogels with high strength, freeze-resistance, and self-adhesive ability. Multiple hydrogen bond networks and ion crosslinking networks are constructed within the hydrogel electrolyte (PVA/PAAc/XG) containing polyvinyl alcohol (PVA), acrylic acid (AAc), and xanthan gum (XG), promoting the enhanced mechanical property, and the adhesion to electrode materials is also improved through abundant active groups. The introduction of zinc ions provides the material with superior frost resistance while also promoting electrical conductivity. Leveraging its multifunction of superior mechanical strength, anti-freeze property, and self-adhesive characteristic, the PVA/PAAc/XG hydrogel electrolyte is employed to fabricate zinc ion hybrid supercapacitors (ZHS). Remarkably, ZHS exhibits outstanding electrochemical performance and cycle stability. A remarkable capacity retention rate of 83.86 % after 10,000 charge-discharge cycles can be achieved at high current densities, even when the operational temperature decreases to -60 °C, showing great potential in the field of flexible energy storage devices.


Assuntos
Polissacarídeos Bacterianos , Cimentos de Resina , Zinco , Hidrogéis , Eletrólitos , Íons
9.
Vaccine ; 42(11): 2781-2792, 2024 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-38508928

RESUMO

Investigating the mechanisms by which W135 meningococcal conjugate (PSW135-TT) activates adaptive immune responses in mice can provide a comprehensive understanding of the immune mechanisms of bacterial polysaccharide conjugate vaccines. We compared B-cell and T-cell immune responses immunized with W135 meningococcal capsular polysaccharides (PSW135), tetanus toxoid (TT) and PSW135-TT in mice. The results showed that PSW135-TT could induce higher PSW135-specific and TT-specific IgG antibodies with a significant enhancement after two doses. All serum antibodies immunized with PSW135- TT had strong bactericidal activity, whereas none of the serum antibodies immunized with PSW135 had bactericidal activity. Besides, IgM and IgG antibodies immunized with PSW135-TT after two doses were positively correlated with the titer of bactericidal antibodies. We also found Th cells favored Th2 humoral immune responses in PSW135-TT, PSW135, and TT-immunized mice, especially peripheral blood lymphocytes. Furthermore, PSW135-TT and TT could effectively activate bone marrow derived dendritic cells (BMDCs) and promote BMDCs to highly express major histocompatibility complex Ⅱ (MHCⅡ), CD86 and CD40 molecules in mice, whereas PSW135 couldn't. These data verified the typical characteristics of PSW135-TT and TT as T cell dependent antigen (TD-Ag) and PSW135 as T cell independent antigen (TI-Ag), which will be very helpful for further exploration of the immune mechanism of polysaccharide-protein conjugate vaccines and improvement of the quality of bacterial polysaccharide conjugate vaccines in future.


Assuntos
Infecções Meningocócicas , Vacinas Meningocócicas , Neisseria meningitidis Sorogrupo W-135 , Animais , Camundongos , Sorogrupo , Toxoide Tetânico , Polissacarídeos Bacterianos , Vacinas Conjugadas , Anticorpos Antibacterianos , Imunidade Celular , Imunoglobulina G , Infecções Meningocócicas/prevenção & controle
10.
Cell ; 187(8): 1874-1888.e14, 2024 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-38518773

RESUMO

Infections of the lung cause observable sickness thought to be secondary to inflammation. Signs of sickness are crucial to alert others via behavioral-immune responses to limit contact with contagious individuals. Gram-negative bacteria produce exopolysaccharide (EPS) that provides microbial protection; however, the impact of EPS on sickness remains uncertain. Using genome-engineered Pseudomonas aeruginosa (P. aeruginosa) strains, we compared EPS-producers versus non-producers and a virulent Escherichia coli (E. coli) lung infection model in male and female mice. EPS-negative P. aeruginosa and virulent E. coli infection caused severe sickness, behavioral alterations, inflammation, and hypothermia mediated by TLR4 detection of the exposed lipopolysaccharide (LPS) in lung TRPV1+ sensory neurons. However, inflammation did not account for sickness. Stimulation of lung nociceptors induced acute stress responses in the paraventricular hypothalamic nuclei by activating corticotropin-releasing hormone neurons responsible for sickness behavior and hypothermia. Thus, EPS-producing biofilm pathogens evade initiating a lung-brain sensory neuronal response that results in sickness.


Assuntos
Infecções por Escherichia coli , Escherichia coli , Pulmão , Polissacarídeos Bacterianos , Infecções por Pseudomonas , Pseudomonas aeruginosa , Animais , Feminino , Masculino , Camundongos , Biofilmes , Escherichia coli/fisiologia , Hipotermia/metabolismo , Hipotermia/patologia , Inflamação/metabolismo , Inflamação/patologia , Pulmão/microbiologia , Pulmão/patologia , Pneumonia/microbiologia , Pneumonia/patologia , Pseudomonas aeruginosa/fisiologia , Células Receptoras Sensoriais , Polissacarídeos Bacterianos/metabolismo , Infecções por Escherichia coli/metabolismo , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Infecções por Pseudomonas/metabolismo , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/patologia , Nociceptores/metabolismo
11.
Carbohydr Polym ; 334: 122008, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38553201

RESUMO

Gellan gum (GG) has attracted considerable attention as a versatile biopolymer with numerous potential biological applications, especially in the fields of tissue engineering, wound healing, and cargo delivery. Due to its distinctive characteristics like biocompatibility, biodegradability, nontoxicity, and gel-forming ability, GG is well-suited for these applications. This review focuses on recent research on GG-based hydrogels and biocomposites and their biomedical applications. It discusses the incorporation of GG into hydrogels for controlled drug release, its role in promoting wound healing processes, and its potential in tissue engineering for various tissues including bone, retina, cartilage, vascular, adipose, and cardiac tissue. It provides an in-depth analysis of the latest findings and advancements in these areas, making it a valuable resource for researchers and professionals in these fields.


Assuntos
Cartilagem , Engenharia Tecidual , Cartilagem/metabolismo , Osso e Ossos , Polissacarídeos Bacterianos/metabolismo , Hidrogéis/farmacologia , Hidrogéis/metabolismo
12.
Carbohydr Res ; 538: 109095, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38507941

RESUMO

Moraxella nonliquefaciens is a commensal of the human upper respiratory tract (URT) but on rare occasions is recovered in cases of ocular, septic and pulmonary infections. Hence there is interest in the pathogenic determinants of M. nonliquefaciens, of which outer membrane (OM) structures such as fimbriae and two capsular polysaccharide (CPS) structures, →3)-ß-D-GalpNAc-(1→5)-ß-Kdop-(2→ and →8)-α-NeuAc-(2→, have been reported in the literature. To further characterise its surface virulence factors, we isolated a novel CPS from M. nonliquefaciens type strain CCUG 348T. This structure was elucidated using NMR data obtained from CPS samples that were subjected to various degrees of mild acid hydrolysis. Together with GLC-MS data, the structure was resolved as a linear polymer composed of two GalfNAc residues consecutively added to Kdo, →3)-ß-D-GalfNAc-(1→3)-α-D-GalfNAc-(1→5)-α-(8-OAc)Kdop-(2→. Supporting evidence for this material being CPS was drawn from the proposed CPS biosynthetic locus which encoded a potential GalfNAc transferase, a UDP-GalpNAc mutase for UDP-GalfNAc production and a putative CPS polymerase with predicted GalfNAc and Kdo transferase domains. This study describes a unique CPS composition reported in Moraxella spp. and offers genetic insights into the synthesis and expression of GalfNAc residues, which are rare in bacterial OM glycans.


Assuntos
Moraxella , Polissacarídeos , Humanos , Polissacarídeos/análise , Transferases/análise , Difosfato de Uridina/análise , Cápsulas Bacterianas/química , Polissacarídeos Bacterianos/química
13.
Carbohydr Res ; 538: 109097, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38555658

RESUMO

The structure of the K141 type capsular polysaccharide (CPS) produced by Acinetobacter baumannii KZ1106, a clinical isolate recovered from Kazakhstan in 2016, was established by sugar analyses and one- and two-dimensional 1H and 13C NMR spectroscopy. The CPS was shown to consist of branched tetrasaccharide repeating units (K-units) with the following structure: This structure was found to be consistent with the genetic content of the KL141 CPS biosynthesis gene cluster at the chromosomal K locus in the KZ1106 whole genome sequence. Assignment of the encoded enzymes allowed the first sugar of the K unit to be identified, which revealed that the ß-d-GlcpNAc-(1→3)-d-GlcpNAc bond is the linkage between K-units formed by the WzyKL141 polymerase.


Assuntos
Acinetobacter baumannii , Acinetobacter baumannii/genética , Acinetobacter baumannii/química , Cápsulas Bacterianas/química , Polissacarídeos/análise , Espectroscopia de Ressonância Magnética , Família Multigênica , Açúcares , Polissacarídeos Bacterianos/química
14.
Carbohydr Res ; 538: 109070, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38460462

RESUMO

This review explores the realm of structural modifications and broad spectrum of their potential applications, with a special focus on the synthesis of xanthan gum derivatives through graft copolymerization methods. It delves into the creation of these derivatives by attaching functional groups (-OH and -COOH) to xanthan gum, utilizing a variety of initiators for grafting, and examining their diverse applications, especially in the areas of food packaging, pharmaceuticals, wastewater treatment, and antimicrobial activities. Xanthan gum is a biocompatible, biodegradable, less toxic, bioactive, and cost-effective natural polymer derived from Xanthomonas species. The native properties of xanthan gum can be improved by cross-linking, grafting, curing, blending, and various modification techniques. Grafted xanthan gum has excellent biodegradability, metal binding, dye adsorption, immunological properties, and wound healing ability. Owing to its remarkable properties, such as biocompatibility and its ability to form gels resembling the extracellular matrix of tissues, modified xanthan gum finds extensive utility across biomedicine, engineering, and the food industry. Furthermore, the review also covers various modified derivatives of xanthan gum that exhibit excellent biodegradability, metal binding, dye adsorption, immunological properties, and wound healing abilities. These applications could serve as important resources for a wide range of industries in future product development.


Assuntos
Polímeros , Polissacarídeos Bacterianos , Polissacarídeos Bacterianos/química , Géis
15.
Carbohydr Polym ; 332: 121889, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38431406

RESUMO

Metabolic alterations within mitochondria, encompassing processes such as autophagy and energy metabolism, play a pivotal role in facilitating the swift proliferation, invasion, and metastasis of cancer cells. Despite this, there is a scarcity of currently available medications with proven anticancer efficacy through the modulation of mitochondrial dysfunction in a clinical setting. Here, we introduce the structural characteristics of RN0D, a galactoglucan isolated and purified from Panax notoginseng flowers, mainly composed of ß-1,4-galactan and ß-1,3/1,6-glucan. RN0D demonstrates the capacity to induce mitochondrial impairment in cancer cells, leading to the accumulation of reactive oxygen species, initiation of mitophagy, and reduction in both mitochondrial number and size. This sequence of events ultimately results in the inhibition of mitochondrial and glycolytic bioenergetics, culminating in the demise of cancer cells due to adenosine triphosphate (ATP) deprivation. Notably, the observed bioactivity is attributed to RN0D's direct targeting of Galectin-3, as affirmed by surface plasmon resonance studies. Furthermore, RN0D is identified as an activator of the PTEN-induced kinase 1 (PINK1)/Parkin pathway, ultimately instigating cytotoxic mitophagy in tumor cells. This comprehensive study substantiates the rationale for advancing RN0D as a potentially efficacious anticancer therapeutic.


Assuntos
Neoplasias , Panax notoginseng , Polissacarídeos Bacterianos , Humanos , Mitofagia , Galactanos , Glucanos , Morte Celular , Ubiquitina-Proteína Ligases/metabolismo , Proteínas Quinases/metabolismo
16.
Appl Environ Microbiol ; 90(4): e0119723, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38551353

RESUMO

Klebsiella pneumoniae, especially hypervirulent K. pneumoniae (hvKP), is a common opportunistic pathogen that often causes hospital- and community-acquired infections. Capsular polysaccharide (CPS) is an important virulence factor of K. pneumoniae. Some phages encode depolymerases that can recognize and degrade bacterial polysaccharides. In this study, the lytic bacteriophage vB_KpnP_ZK1 (abbreviated as ZK1) was isolated using serotype K1 hvKP as the host. Although amino acid sequence BLAST analysis indicated that the tail fiber protein Depo16 of phage ZK1 showed no significant similarity to any reported phage depolymerases, it displayed enzymatic activities that are characteristic of phage depolymerases. After expression and purification, Depo16 could efficiently remove the capsular polysaccharide layer that surrounds the surface of serotype K1 K. pneumoniae. Although no bactericidal activity was detected, Depo16 makes serotype K1 K. pneumoniae sensitive to peritoneal macrophages (PMs). In addition, in a mouse bacteremia model of serotype K1 K. pneumoniae, 25 µg of Depo16 was effective in significantly prolonging survival. Depo16 treatment can reduce the bacterial load in blood and major tissues and alleviate tissue damage in mice. This indicates that the putative depolymerase Depo16 is a potential antibacterial agent against serotype K1 K. pneumoniae infections.IMPORTANCEKlebsiella pneumoniae often causes hospital-acquired infections and community-acquired infections. Capsular polysaccharide (CPS) is one of the crucial virulence factors of K. pneumoniae. K1 and K2 capsular-type K. pneumoniae strains are the most prevalent serotypes of hypervirulent K. pneumoniae (hvKP). In this study, a novel K. pneumoniae phage named vB_KpnP_ZK1 was isolated, and its putative depolymerase Depo16 showed low homology with other reported phage depolymerases. Depo16 can specifically degrade the K. pneumoniae K1 capsule making this serotype sensitive to peritoneal macrophages. More importantly, Depo16 showed a significant therapeutic effect in a mouse bacteremia model caused by serotype K1 K. pneumoniae. Thus, Depo16 is a potential antibacterial agent to combat serotype K1 K. pneumoniae infections.


Assuntos
Bacteriemia , Bacteriófagos , Infecções Comunitárias Adquiridas , Infecções por Klebsiella , Animais , Camundongos , Klebsiella pneumoniae , Bacteriófagos/genética , Infecções por Klebsiella/terapia , Infecções por Klebsiella/microbiologia , Fatores de Virulência/metabolismo , Polissacarídeos Bacterianos , Antibacterianos
17.
Int J Biol Macromol ; 264(Pt 1): 130604, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38447843

RESUMO

Skin, the largest organ in the body, blocks the entry of environmental pollutants into the system. Any injury to this organ allows infections and other harmful substances into the body. 3D bioprinting, a state-of-the-art technique, is suitable for fabricating cell culture scaffolds to heal chronic wounds rapidly. This study uses starch extracted from Maranta arundinacea (Arrowroot plant) (AS) and gellan gum (GG) to develop a bioink for 3D printing a scaffold capable of hosting animal cells. Field emission scanning electron microscopy (FE-SEM) and X-ray diffraction analysis (XRD) prove that the isolated AS is analogous to commercial starch. The cell culture scaffolds developed are superior to the existing monolayer culture. Infrared microscopy shows the AS-GG interaction and elucidates the mechanism of hydrogel formation. The physicochemical properties of the 3D-printed scaffold are analyzed to check the cell adhesion and growth; SEM images have confirmed that the AS-GG printed scaffold can support cell growth and proliferation, and the MTT assay shows good cell viability. Cell behavioral and migration studies reveal that cells are healthy. Since the scaffold is biocompatible, it can be 3D printed to any shape and structure and will biodegrade in the requisite time.


Assuntos
Marantaceae , Polissacarídeos Bacterianos , Tecidos Suporte , Animais , Tecidos Suporte/química , Amido , Cicatrização , Impressão Tridimensional , Engenharia Tecidual , Hidrogéis/farmacologia
18.
ACS Appl Mater Interfaces ; 16(12): 14573-14582, 2024 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-38484043

RESUMO

Pseudomonas aeruginosa biofilms comprise three main polysaccharides: alginate, psl, and pel, which all imbue tolerance against exogenous antimicrobials. Nanoparticles (NPs) are an exciting new strategy to overcome the biofilm matrix for therapeutic delivery applications; however, zero existing FDA approvals for biofilm-specific NP formulations can be attributed to the complex interplay of physiochemical forces at the biofilm-NP interface. Here, we leverage a set of inducible, polysaccharide-specific, expressing isogenic P. aeruginosa mutants coupled with an assembled layer-by-layer NP (LbL NP) panel to characterize biofilm-NP interactions. When investigating these interactions using confocal microscopy, alginate-layered NPs associated more than dextran-sulfate-layered NPs with biofilms that had increased alginate production, including biofilms produced by mucoid P. aeruginosa isolates from people with cystic fibrosis. These differences were further confirmed in LbL NPs layered with polysaccharide- or hydrocarbon-based polymers with pendent carboxylate or sulfate functional groups. These data suggest carboxylated NP surfaces have enhanced interactions specifically with mucoid biofilms as compared to sulfated surfaces and lay the foundation for their inclusion as a design element for increasing biofilm-NP interactions and efficacious drug delivery.


Assuntos
Nanopartículas , Pseudomonas aeruginosa , Humanos , Polissacarídeos Bacterianos , Biofilmes , Ácidos Carboxílicos , Alginatos , Sulfatos
19.
Int J Biol Macromol ; 265(Pt 1): 130864, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38493820

RESUMO

Ketoprofen (KET), commonly used for inflammation in clinical settings, leads to systemic adverse effects with prolonged use, mitigated by topical administration. Nanotechnology-based cutaneous forms, like films, may enhance KET efficacy. Therefore, this study aimed to prepare and characterize films containing KET nanoemulsions (F-NK) regarding mechanical properties, chemical composition and interactions, occlusive potential, bioadhesion, drug permeation in human skin, and safety. The films were prepared using a κ-carrageenan and xanthan gum blend (2 % w/w, ratio 3: 1) plasticized with glycerol through the solvent casting method. Non-nanoemulsioned KET films (F-K) were prepared for comparative purposes. F-NK was flexible and hydrophilic, exhibited higher drug content and better uniformity (94.40 ± 3.61 %), maintained the NK droplet size (157 ± 12 nm), and was thinner and lighter than the F-K. This film also showed increased tensile strength and Young's modulus values, enhanced bioadhesion and occlusive potential, and resulted in more of the drug in the human skin layers. Data also suggested that nano-based formulations are homogeneous and more stable than F-KET. Hemolysis and chorioallantoic membrane tests suggested the formulations' safety. Thus, the nano-based film is suitable for cutaneous KET delivery, which may improve the drug's efficacy in managing inflammatory conditions.


Assuntos
Cetoprofeno , Nanocompostos , Polissacarídeos Bacterianos , Humanos , Cetoprofeno/farmacologia , Cetoprofeno/química , Carragenina/química , Pele , Nanocompostos/química
20.
Int J Biol Macromol ; 264(Pt 2): 130572, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38447825

RESUMO

To effectively extend the shelf life of fruits meanwhile facilitating consumers to judge their freshness, in this work, a double-layer multifunctional film combining CO2 sensitivity and antibacterial properties was successfully prepared by adding methyl red (MR), bromothymol blue (BTB) into gellan gum (GG) as the sensing inner layer, and doping tannic acid (TA) into sodium alginate with sodium carboxymethyl cellulose (CMC) as the antimicrobial outer layer, which was applied to the freshness indication of strawberries. Microscopic morphology and spectral analysis demonstrated that the bi-layer films were fabricated successfully. The mechanical characteristics, thermal stability, water vapor resistance, and antibacterial capabilities of the bilayer films improved as TA concentration rose. They exhibited noticeable color changes at pH = 2-10 and different concentrations of CO2. Application of the prepared films to strawberries revealed that the GG-MB@SC-6%TA film performed most favorably under 4 °C storage conditions, not only monitoring strawberry freshness but also retaining high soluble solids and titratable acidity, resulting in a slight decrease in hardness and weight loss. Therefore, taking into account all of the physical-functional characteristics, the GG-MB@6%TA film has a broad application prospect for intelligent food packaging.


Assuntos
Anti-Infecciosos , Fragaria , Polifenóis , Polissacarídeos Bacterianos , Dióxido de Carbono , Carboximetilcelulose Sódica/química , Alginatos , Antibacterianos/farmacologia , Antibacterianos/química , Embalagem de Alimentos/métodos , Sódio
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