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The effective treatment of chronic wounds represents a critical global medical challenge demanding urgent attention. Persistent inflammation, driven by an excess of reactive oxygen radicals, sets in motion a detrimental cycle leading to chronic wounds and impeding the natural healing process. This study develops a sprayable wound dressing by covalently grafting amino fullerene to carboxymethylated curdlan (CMC-C). This novel dressing exhibits excellent biocompatibility, antioxidant, and reactive oxygen species scavenging properties. Furthermore, it demonstrates a targeted affinity for HEK-a cells, efficiently reducing the inflammatory response while promoting cell proliferation and migration in vitro. Moreover, the animal experiment investigations reveal that CMC-C significantly accelerates chronic wounds healing by regulating the inflammatory process, promoting collagen deposition, and improving vascularization. These results demonstrate the potential of the sprayable dressing (CMC-C) in curing the healing of chronic wounds through the modulation of the inflammatory microenvironment. Overall, the sprayable hydrogel dressing based on water-soluble derivative of fullerene and curdlan emerges as a potential approach for clinical applications in the treatment of chronic wounds.
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BACKGROUND: Laboratory scale experiments have shown that curdlan and gellan gum gelled together as curdlan/gellan gum (CG) hybrid gels showed better gel properties than the individual curdlan and gellan gum. In this study, CG and black wolfberry anthocyanin (BWA), CG and maltitol (ML) hybrid gels were constructed using CG hybrid gel as matrix. The effects of BWA or ML on the gel properties and microstructure of CG hybrid gels were investigated and a confectionery gel was developed. RESULTS: The presence of BWA increased the storage modulus (G') value of CG at 0.1 Hz, whereas ML had little effect on the G' value of CG. The addition of BWA (5 g L-1 ) and ML (0.3 mol L-1 ) increased the melting and gelling temperatures of CG hybrid gels to 42.4 °C and 34.1 °C and 44.2 °C and 33.2 °C, respectively. Meanwhile, the relaxation time T22 in CG-ML and CG-BWA hybrid gels was reduced to 91.96 and 410.27 ms, indicating the strong binding between BWA and CG, ML and CG. The hydrogen bond interaction between BWA or ML and CG was confirmed by the shift in the hydroxyl stretching vibration peak. Moreover, the microstructures of CG-ML and CG-BWA hybrid gels were denser than that of CG. In addition, confectionery gel containing CG-BWA-ML has good chewing properties. CONCLUSION: These results indicated that the incorporation of BWA or ML could improve the structure of CG hybrid gels and assign a sustainability potential for the development of confectionery gels based on CG complex. © 2024 Society of Chemical Industry.
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Lycium , Maltose/análogos & derivados , Álcoois Açúcares , beta-Glucanas , Antocianinas , Polissacarídeos Bacterianos/química , Géis/química , ReologiaRESUMO
Curdlan is an edible microbial polysaccharide and can be used in food, biomedical and biomaterial fields. To reduce the cost of curdlan production, this study investigated the suitability of cassava starch hydrolysates as carbon source for curdlan production. Cassava starch was hydrolyzed into maltose syrup using ß-amylase and pullulanase at various enzyme dosages, temperature, time and addition order of two enzymes. The maltose yield of 53.17% was achieved at starch loading 30% by simultaneous addition ß-amylase 210 U/g starch and pullulanase 3 U/g starch at 60 °C for 9 h. Cassava starch hydrolysates were used as carbon source for curdlan production by Agrobacterium sp. DH-2. The curdlan production reached 28.4 g/L with the yield of 0.79 g/g consumed sugar and molecular weight of 1.26 × 106 Da at 96 h with cassava starch hydrolysate at 90 g/L initial sugar concentration. Curdlan produced from cassava starch hydrolysates was characterized using FT-IR spectra and thermo gravimetric analysis. This work indicated that cassava starch was a potential renewable feedstock for curdlan production.
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Manihot , beta-Amilase , Agrobacterium , Carbono , Maltose , Espectroscopia de Infravermelho com Transformada de Fourier , Amido , beta-GlucanasRESUMO
Mast cells (MCs) are engaged in host defense against various pathogens as they are equipped with pattern recognition receptors (PRRs). Among PRRs expressed on MCs, there are also molecules recognizing components of the fungal cell wall, which are able to induce cellular activation and response. However, little information is available concerning the MC activation by various fungal-derived components. The aim of the study was to determine whether curdlan, a model fungal particle of ß-(1,3)-glucan, can directly stimulate tissue MCs. We demonstrated that curdlan triggers MCs to initiate pro-inflammatory response as it activates these cells to synthesize essential pro-inflammatory and/or immunoregulatory factors. We also showed that curdlan serves as a potent chemoattractant for MCs and stimulates those cells to generate reactive oxygen species (ROS). Finally, we documented that curdlan induces MC response via Dectin-1. Our observations support the idea that MCs serve as important sentinels modulating immune response during fungal infection.
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Degranulação Celular/imunologia , Quimiotaxia de Leucócito/imunologia , Lectinas Tipo C/imunologia , Mastócitos/imunologia , beta-Glucanas/imunologia , Animais , Feminino , Ratos , Ratos Wistar , Espécies Reativas de OxigênioRESUMO
Eugenol is hepatotoxic and potentially hazardous to human health. This paper reports on a rapid non-destructive quantitative method for the determination of eugenol concentration in curdlan (CD) biofilms by electronic nose (E-nose) combined with gas chromatography-mass spectrometry (GC-MS). Different concentrations of eugenol were added to the film-forming solution to form a series of biofilms by casting method, and the actual eugenol concentration in the biofilm was determined. Analysis of the odor collected on the biofilms was carried out by GC-MS and an E-nose. The E-nose data was subjected to principal component analysis (PCA) and linear discriminant analysis (LDA) in order to establish a discriminant model for determining eugenol concentrations in the biofilms. Further analyses involving the application of all sensors and featured sensors, the prediction model-based partial least squares (PLS) and support vector machines (SVM) were carried out to determine eugenol concentration in the CD biofilms. The results showed that the optimal prediction model for eugenol concentration was obtained by PLS at R2p of 0.952 using 10 sensors. The study described a rapid, non-destructive detection and quantitative method for determining eugenol concentration in bio-based packaging materials.
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Biofilmes , Nariz Eletrônico , Cromatografia Gasosa-Espectrometria de Massas , Eugenol , Humanos , Odorantes/análise , beta-GlucanasRESUMO
Low water solubility frequently compromises the therapeutic efficacy of drugs and other biologically active molecules. Here, we report on coacervate polysaccharide nanoparticles (CPNs) that can transport and release a model hydrophobic drug, piroxicam, to the cells in response to changes in temperature. The proposed, temperature-responsive drug delivery system is based on ionic derivatives of natural polysaccharides-curdlan and hydroxypropyl cellulose. Curdlan was modified with trimethylammonium groups, while the anionic derivative of hydroxypropyl cellulose was obtained by the introduction of styrenesulfonate groups. Thermally responsive nanoparticles of spherical shape and average hydrodynamic diameter in the range of 250-300 nm were spontaneously formed in water from the obtained ionic polysaccharides as a result of the coacervation process. Their morphology was visualized using SEM and AFM. The size and the surface charge of the obtained objects could be tailored by adjusting the polycation/polyanion ratio. Piroxicam (PIX) was effectively entrapped inside the nanoparticles. The release profile of the drug from the CPNs-PIX was found to be temperature-dependent in the range relevant for biomedical applications.
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Anti-Inflamatórios não Esteroides/administração & dosagem , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Piroxicam/administração & dosagem , Polissacarídeos/química , Algoritmos , Celulose/análogos & derivados , Celulose/química , Técnicas de Química Sintética , Microscopia de Força Atômica , Modelos Teóricos , Estrutura Molecular , Polieletrólitos/química , Análise Espectral , TemperaturaRESUMO
Curdlan has wide potential application in the food and biomedical fields due to its unique thermal gel and biological activity. This study investigated the effect of six sugars including glucose, fructose, lactose, maltose, sucrose and xylose as carbon sources on production and properties of curdlan using Agrobacterium sp. DH-2. The maximum production (38.1 g/L and 37.4 g/L, respectively) and yield (0.58 g curdlan/g sucrose and 0.53 g curdlan/g maltose, respectively) of curdlan were achieved by sucrose and maltose, followed by glucose, fructose, lactose and xylose. Scanning electron micrographs showed that the surface of cells was smooth in strain growth phase, while cells were covered by curdlan matrix acted as a net in the curdlan synthesis phase. The highest glucosyltransferase activity (19.9 U/g biomass) corresponded to the maximum curdlan production using the sucrose medium. The molecular weight and gel strength of curdlan were influenced by the carbon sources. The curdlan from xylose medium resulted in a maximum molecular weight of 1.59 × 106 Da and the highest gel strength of 989.2 g/cm2, while the curdlan from sucrose medium resulted in a lowest molecular weight of 1.10 × 106 Da and gel strength of 672.8 g/cm2. The high molecular weight of curdlan had high gel strength.
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Agrobacterium/metabolismo , Microbiologia Industrial , beta-Glucanas/metabolismo , Agrobacterium/enzimologia , Carbono/metabolismo , Meios de Cultura/metabolismo , Glucosiltransferases/metabolismo , Maltose/metabolismo , Sacarose/metabolismoRESUMO
Resolution of inflammation that occurs after microbial infection or tissue damage is an important physiologic process in maintaining or restoring host homeostasis. Taurine chloramine (TauCl) is formed by a reaction between taurine and hypochlorite in leukocytes, and it is especially abundant in activated neutrophils that encounter an oxidative burst. As neutrophils undergo apoptosis, TauCl is released to the extracellular matrix at the inflamed sites, thereby affecting coexisting macrophages in the inflammatory microenvironment. In this study, we investigated the role of TauCl in phagocytosis by macrophages during resolution of fungal infection-induced inflammation. We found that exogenous TauCl substantially increased the phagocytic efficiency of macrophages through up-regulation of dectin-1, a receptor for fungal ß-1,3-glucans, which is present on the membrane of macrophages. Our previous studies demonstrated the induction of heme oxygenase-1 (HO-1) expression in murine peritoneal macrophages treated with TauCl. In the present study, knocking out HO-1 or pharmacologic inhibition of HO-1 with zinc protoporphyrin IX attenuated the TauCl-induced expression of dectin-1 and subsequent phagocytosis. Furthermore, carbon monoxide (CO), a by-product of the HO-1-catalyzed reaction, induced expression of dectin-1 and potentiated phagocytic capability of the macrophages, which appeared to be mediated through up-regulation of peroxisome proliferator-activated receptor γ. Taken together, induction of HO-1 expression and subsequent CO production by TauCl are essential for phagocytosis of fungi by macrophages. Our results suggest that TauCl has important roles in host defense against fungal infection and has therapeutic potential in the management of inflammatory diseases.-Kim, S. H., Zhong, X., Kim, W., Kim, K., Suh, Y.-G., Kim, C., Joe, Y., Chung, H. T., Cha, Y.-N., Surh, Y.-J. Taurine chloramine potentiates phagocytic activity of peritoneal macrophages through up-regulation of dectin-1 mediated by heme oxygenase-1-derived carbon monoxide.
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Inibidores Enzimáticos/farmacologia , Lectinas Tipo C/metabolismo , Macrófagos Peritoneais/efeitos dos fármacos , Fagocitose , Taurina/análogos & derivados , Regulação para Cima , Animais , Candida albicans/patogenicidade , Monóxido de Carbono/metabolismo , Células Cultivadas , Heme Oxigenase-1/antagonistas & inibidores , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Lectinas Tipo C/genética , Macrófagos Peritoneais/metabolismo , Macrófagos Peritoneais/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , PPAR gama , Taurina/farmacologiaRESUMO
Functional oligosaccharides, particularly curdlan (1â3)-ß-d-glucan oligosaccharides (GOS), play important roles in modulating host immune responses. However, the molecular mechanisms underlying the immunostimulatory effects of GOS on macrophage polarization are not clear. In this work, GOS (5-1000 µg/mL) were non-toxic to bone marrow-derived macrophages (BMDMs) with improved pinocytic and bactericidal capacities. Incubation with GOS (100 µg/mL) induced M1 phenotype polarization of BMDMs as evidenced by increased CD11c+/CD86+ (10.1%) and M1 gene expression of inducible nitric oxide synthase, interleukin (IL)-1ß, and chemokine C-C-motif ligand 2. Accordingly, the secretion of cytokines IL-1ß, IL-6, monocyte chemotactic protein-1, and tumor necrosis factor-α, as well as the nitrite release of BMDMs were increased by GOS (100 µg/mL). Expression of mitogen-activated protein kinases (MAPKs) of phosphorylated (p)-c-Jun amino-terminal kinase, p-extracellular signal regulated kinase, and p-p38 in BMDMs were increased by GOS, as well as the p-Stat1. Moreover, nuclear factor-kappa B (NF-κB) p-p65 expression in BMDMs was promoted by GOS while it suppressed IκBα expression. Receptor blocking with anti-CR3 (CD11b/CD18) and anti-toll-like receptor (TLR) 2 antibodies diminished GOS induced M1 phenotype polarization with reduced mRNA expression of M1 genes, decreased cytokine and nitrite releases, and suppressed signaling pathway activation. Thus, CR3 (CD11b/CD18) and TLR2 mediated activation of MAPKs and NF-κB pathways are responsible for GOS induced polarization of BMDMs.
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Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Macrófagos/citologia , NF-kappa B/metabolismo , Oligossacarídeos/farmacologia , beta-Glucanas/química , Alcaligenes faecalis , Animais , Polaridade Celular/efeitos dos fármacos , Proliferação de Células , Células Cultivadas , Citocinas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Oligossacarídeos/síntese química , Oligossacarídeos/química , Fosforilação , Pinocitose/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , beta-Glucanas/farmacologiaRESUMO
OBJECTIVE: Curdlan has been widely used as a gelling agent in various food systems. This study was performed to evaluate the rheological properties of pork myofibrillar protein (MP) with different levels of curdlan (0.5% to 1.5%) and its application to low-fat model sausages (LFS). METHODS: MP mixtures were prepared with 0.5%, 1.0%, and 1.5% of curdlan. Cooking loss (%), gel strength (gf), shear stress (Pa), and scanning electron microscopy were measured. Physicochemical and textural properties of LFS containing different levels of curdlan were measured. RESULTS: The shear stress of MP mixtures increased with increasing levels of curdlan. MP gels with increased levels of curdlan decreased cooking loss and increased gel strength (p<0.05). The MPs with 1.0% and 1.5% of curdlan were observed more compact three-dimensional structure than those with 0.5% curdlan. Increased curdlan level in LFS affected redness (a*) and yellowness (b*) values. Although expressible moisture of LFS did not differ among curdlan levels, LFSs with various levels of curdlan decreased cooking loss as compared to control sausages. Hardness values (2,251 to 2,311 gf) of LFS with 0.5% and 1.0% curdlan was increased and differ from those (1,901 gf) of control sausages. CONCLUSION: The addition of 1.0% curdlan improved the functional and textural properties of LFS.
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In this study, to improve the thermal and mechanical properties of chitosan films, a chitosan/curdlan/carboxymethyl cellulose (CS/CD/CMC) ternary blended film was prepared and characterized. To prepare a uniform CS/CD/CMC ternary blended film, an effective method of blending CD with other materials was established as the following conditions: the ternary solution temperature was maintained at 60 °C, and the pH was controlled in the range from 12 to 4. Compared to the pure chitosan, the CS/CD/CMC blended films exhibited better mechanical properties, permeability, and thermal stability. In addition, visible light properties of the ternary blending film were improved. Scanning electron microscope and Fourier transform-infrared spectroscopy analyses indicated good compatibility among the CS, CD and CMC, which led to a corresponding improvement in the properties owing to interactions among the three components in the blending process. So, an effective method of blending CD with CS and CMC was established, and the blending film has good thermal and mechanical properties.
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BH0236 from Bacillus halodurans is a multimodular ß-1,3-glucanase comprising an N-terminal family 81 glycoside hydrolase catalytic module, an internal family 6 carbohydrate-binding module (CBM) that binds the nonreducing end of ß-1,3-glucan chains, and an uncharacterized C-terminal module classified into CBM family 56. Here, we determined that this latter CBM, BhCBM56, bound the soluble ß-1,3-glucan laminarin with a dissociation constant (Kd ) of â¼26 µm and displayed higher affinity for insoluble ß-1,3-glucans with Kd values of â¼2-10 µm but lacked affinity for ß-1,3-glucooligosaccharides. The X-ray crystal structure of BhCBM56 and NMR-derived chemical shift mapping of the binding site revealed a ß-sandwich fold, with the face of one ß-sheet possessing the ß-1,3-glucan-binding surface. On the basis of the functional and structural properties of BhCBM56, we propose that it binds a quaternary polysaccharide structure, most likely the triple helix adopted by polymerized ß-1,3-glucans. Consistent with the BhCBM56 and BhCBM6/56 binding profiles, deletion of the CBM56 from BH0236 decreased activity of the enzyme on the insoluble ß-1,3-glucan curdlan but not on soluble laminarin; additional deletion of the CBM6 also did not affect laminarin degradation but further decreased curdlan hydrolysis. The pseudo-atomic solution structure of BH0236 determined by small-angle X-ray scattering revealed structural insights into the nature of avid binding by the BhCBM6/56 pair and how the orientation of the active site in the catalytic module factors into recognition and degradation of ß-1,3-glucans. Our findings reinforce the notion that catalytic modules and their cognate CBMs have complementary specificities, including targeting of polysaccharide quaternary structure.
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Bacillus/enzimologia , Proteínas de Bactérias/química , Glucana 1,3-beta-Glucosidase/química , Bacillus/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Glucana 1,3-beta-Glucosidase/genética , Glucana 1,3-beta-Glucosidase/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo , Estrutura Secundária de ProteínaRESUMO
Invasion of host cell by pathogens induce various intracellular signalling pathways. The host cell through the initiation of these signalling circuits desperately wants to get rid of the pathogen, whereas the pathogen tries to subvert these defence strategies to create an environment for their successful survival. Leishmania spp. is not an exception. Leishmania have to evolve a range of strategic mechanisms to neutralize macrophage defensive arsenals which enable the parasite to replicate within the phagolysosome of infected host. Understanding these signalling mechanisms in detail will not only improve our basic knowledge of host-pathogen interaction but will also help us to develop effective drug targets not only against leishmaniasis but also for many other macrophage associated diseases. © 2018 IUBMB Life, 70(7):593-601, 2018.
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Antiprotozoários/farmacologia , Interações Hospedeiro-Patógeno/fisiologia , Leishmania donovani/patogenicidade , Leishmaniose Visceral/etiologia , Apoptose/fisiologia , Dinoprostona/imunologia , Dinoprostona/metabolismo , Humanos , Tolerância Imunológica , Fatores Imunológicos/farmacologia , Inflamassomos , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/parasitologia , Explosão Respiratória , Transdução de Sinais , Receptores Toll-Like/imunologia , Receptores Toll-Like/metabolismoRESUMO
BACKGROUND: Streptococcus pneumoniae is a respiratory pathogen causing severe lung infection that may lead to complications such as bacteremia. Current polysaccharide vaccines have limited serotype coverage and therefore cannot provide maximal and long-term protection. Global efforts are being made to develop a conserved protein vaccine candidate. PrtA, a pneumococcal surface protein, was identified by screening a pneumococcal genomic expression library using convalescent patient serum. The prtA gene is prevalent and conserved among S. pneumoniae strains. Its protective efficacy, however, has not been described. Mucosal immunization could sensitize both local and systemic immunity, which would be an ideal scenario for preventing S. pneumoniae infection. METHODS: We immunized BALB/c mice intranasally with a combination of a PrtA fragment (amino acids 144-1041) and Th17 potentiated adjuvant, curdlan. We then measured the T-cell and antibody responses. The protective efficacy conferred to the immunized mice was further evaluated using a murine model of acute pneumococcal pneumonia and pneumococcal bacteremia. RESULTS: There was a profound antigen-specific IL-17A and IFN-γ response in PrtA-immunized mice compared with that of adjuvant control group. Even though PrtA-specific IgG and IgA titer in sera was elevated in immunized mice, only a moderate IgA response was observed in the bronchoalveolar lavage fluid. The PrtA-immunized antisera facilitated the activated murine macrophage, RAW264.7, to opsonophagocytose S. pneumoniae D39 strain; however, PrtA-specific immunoglobulins bound to pneumococcal surfaces with a limited potency. Finally, PrtA-induced immune reactions failed to protect mice against S. pneumoniae-induced acute pneumonia and bacterial propagation through the blood. CONCLUSIONS: Immunization with recombinant PrtA combined with curdlan produced antigen-specific antibodies and elicited IL-17A response. However, it failed to protect the mice against S. pneumoniae-induced infection.
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Proteínas de Bactérias/administração & dosagem , Imunização/métodos , Vacinas Pneumocócicas/administração & dosagem , Pneumonia Pneumocócica/prevenção & controle , Streptococcus pneumoniae , Animais , Proteínas de Bactérias/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fagocitose/efeitos dos fármacos , Fagocitose/fisiologia , Vacinas Pneumocócicas/imunologia , Pneumonia Pneumocócica/imunologia , Pneumonia Pneumocócica/metabolismo , Células RAW 264.7 , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/imunologiaRESUMO
Fungal ß-glucan is a potent immunological stimulator, and that it activates both the innate immune system and adaptive immunity. Curdlan is (1â3)-ß-glucan, a linear form of ß-glucan with a high molecular weight; it modulates the immune response. However, its role in bone tissue is controversial, and the effects of curdlan on bone tissues are unknown. Toll-like receptors (TLRs) play critical roles in innate immunity, and various ligands for TLRs are thought to regulate the host defense mechanisms against pathogens. TLR2 is known to form heterodimers with TLR6, and the TLR2-TLR6 heterodimer (TLR2/6) recognizes diacylated lipopeptides from Gram-positive bacteria. In the present study, we prepared low molecular-weight curdlan, (1â3)-ß-D-glucan, and examined its effects on bone resorption induced by TLR2/6 signaling. In co-cultures of bone marrow cells and osteoblasts, low molecular-weight curdlan suppressed the osteoclast formation induced by TLR2/6 ligand, and attenuated bone resorption in mouse calvarial organ cultures. Curdlan acted on mouse osteoblasts and suppressed the expression of receptor activator of nuclear factor-kappa B (NF-κB) ligand (RANKL), a key molecule for osteoclastogenesis. Curdlan also acted on mouse bone marrow macrophages and suppressed RANKL-dependent osteoclast differentiation from osteoclast precursor cells. The present study indicates that low molecular-weight curdlan attenuated TLR2-induced inflammatory bone resorption. Curdlan, (1â3)-ß-glucan may be a natural agent with beneficial effects on bone health in humans.
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Células da Medula Óssea/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , beta-Glucanas/farmacologia , Animais , Células da Medula Óssea/citologia , Reabsorção Óssea , Células Cultivadas , Técnicas de Cocultura , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteoclastos/citologia , Osteoprotegerina/genética , Ligante RANK/genética , RNA Mensageiro/metabolismo , Crânio/efeitos dos fármacos , Receptor 2 Toll-Like , beta-Glucanas/químicaRESUMO
To optimize the physicochemical properties of phthalocyanine (PC), we examined its behavior in particles of triple helix glucan curdlan (CUR). CUR was denatured and renatured in DMSO, in the presence of PC. Infrared spectroscopy and transmission electron microscopy (TEM) showed that PC and CUR formed an inclusion complex, in which PC was trapped inside CUR molecules. This redshifted the absorption peak of PC, which would improve its usefulness as a photosensitizer, because infrared light can penetrate more deeply into human tissues. The conductivity of the solution of CUR-PC was higher than the conductivities of either a CUR solution or a PC dispersion, indicating that CUR-PC is more water soluble than PC. In addition, CUR-PC was highly stable in water. Thus, the use of CUR as a carrier of PC improves several of its physical properties. PC is used as a photosensitizer for killing cancer cells, but its use is hampered by its low solubility. Further, its absorption range limits its use to a depth of 1â»3 mm in tissues. CUR-PC, with its high solubility and infrared absorption peak, was highly effective as a photosensitizer. It killed 84% of HeLa cells under 15 min of long wavelength radiation and had little cytotoxicity in the absence of light. These results demonstrate that CUR-PC has promise as a photosensitizer, as well as provide theoretical support for a wide range of applications for PC and CUR.
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Indóis/química , Fármacos Fotossensibilizantes/química , beta-Glucanas/química , Células HeLa , Humanos , Isoindóis , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Tamanho da Partícula , Solubilidade , Espectrofotometria Infravermelho , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Curdlan gum is a neutral water-insoluble bacterial exopolysaccharide composed primarily of linear ß-(1,3) glycosidic linkages. Recently, there has been increasing interest in the applications of curdlan and its derivatives. Curdlan is found to inhibit tumors and its sulfated derivative possess anti-HIV activity. Curdlan is biodegradable, non-toxic towards human, environment and edible which makes it suitable as drug-delivery vehicles for sustained drug release. The increasing demand for the growing applications of curdlan requires an efficient high yield fermentation production process so as to satisfy the industrial needs. In this perspective, the present work is aimed to screen and isolate an efficient curdlan gum producing bacteria from rhizosphere of ground nut plant using aniline-blue agar. High yielding isolate was selected based on curdlan yield and identified as Bacillus cereus using gas-chromatography fatty acid methyl ester analysis. B. cereus PR3 curdlan gum was characterized using FT-IR spectroscopy, SEM, XRD and TGA. Fermentation time for curdlan production using B. cereus PR3 was optimized. Media constituents like carbon, nitrogen and mineral sources were screened using Plackett-Burman design. Subsequent statistical analysis revealed that Starch, NH4NO3, K2HPO4, Na2SO4, KH2SO4 and CaCl2 were significant media constituents and these concentrations were optimized for enhancement of curdlan production up to 20.88 g/l.
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Bacillus cereus/metabolismo , Microbiologia Industrial/métodos , Polissacarídeos Bacterianos/metabolismo , beta-Glucanas/metabolismo , Meios de Cultura/análise , Meios de Cultura/metabolismo , Fabaceae/microbiologia , Fermentação , Humanos , RizosferaRESUMO
Water-soluble ß-1,3-glucan (w-glucan) prepared from curdlan is reported to possess various bioactive and medicinal properties. To develop an efficient and cost-effective microbial fermentation method for the direct production of w-glucan, a coupled fermentation system of Agrobacterium sp. and Trichoderma harzianum (CFS-AT) was established. The effects of Tween-80, glucose flow rate, and the use of a dissolved oxygen (DO) control strategy on w-glucan production were assessed. The addition of 10 g L-1 Tween-80 to the CFS-AT enhanced w-glucan production, presumably by loosening the curdlan ultrastructure and increasing the efficiency of curdlan hydrolysis. A two-stage glucose and DO control strategy was optimal for w-glucan production. At the T. harzianum cell growth stage, the optimal glucose flow rate and agitation speed were 2.0 g L-1 hr-1 and 600 rpm, respectively, and at the w-glucan production stage, they were 0.5 g L-1 hr-1 and 400 rpm, respectively. W-glucan production reached 17.31 g L-1, with a degree of polymerization of 19-25. Furthermore, w-glucan at high concentrations exhibited anti-tumor activity against MCF-7, HepG2, and Hela cancer cells in vitro. This study provides a novel, cost-effective, eco-friendly, and efficient microbial fermentation method for the direct production of biologically active w-glucan.
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Agrobacterium/metabolismo , Microbiologia Industrial/métodos , Trichoderma/metabolismo , beta-Glucanas/metabolismo , Agrobacterium/química , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Fermentação , Glucose/metabolismo , Humanos , Hidrólise , Neoplasias/tratamento farmacológico , Oxigênio/metabolismo , Polissorbatos/metabolismo , Solubilidade , Trichoderma/química , Água/química , beta-Glucanas/química , beta-Glucanas/farmacologiaRESUMO
Saccharide sensing in aqueous media is an intriguing but challenging goal in current chemistry. Herein we report the oligosaccharide-sensing behavior of newly synthesized porphyrin-curdlan conjugates, which are random coils in DMSO but become globules in aqueous solutions to induce circular dichroism (ICD) in the biologically accessible spectral region due to the conformational fixation of porphyrin reporters. The magnitude of ICD was significantly varied specifically in the presence of acarbose, a drug for type-2 diabetes, enabling us to detect the aminosaccharide at concentrations down to 200â µm. This result demonstrates that the prêt-á-porter approach, using less-defined reporter-curdlan conjugates, is more advantageous than the traditional haute-couture approach with highly sophisticated hosts in particular in oligosaccharide sensing.
RESUMO
BACKGROUND: This work evaluated the improvement of curdlan production of Agrobacterium sp. ATCC 31749 by using culture medium containing juice of discarded bottom part of green Asparagus spear (MJDA). Curdlan production was carried out using Agrobacterium sp. ATCC 31749 in flasks with different volumes of MJDA and its non-juice-adding control (CK) incubated in shaker at 30 °C, 200 rpm rotation for 168 h. RESULTS: All MJDA media increased Agrobacterium sp. ATCC 31749 cell mass and enhanced the cells' ability to utilise sucrose, the carbon source for curdlan biosynthesis, and thereby produced higher concentration of curdlan than CK which is used for commercial production of curdlan. After 168 h of fermentation, 10% MJDA produced 40.2 g/l of curdlan whiles CK produced 21.1 g/l. Curdlan production was increased by 90.4% higher in 10% MJDA than CK. Curdlan produced by 10% MJDA contains 1.2 and 1.5 µg/ml of Asparagus flavonoids and saponins respectively as additives which have wide range of health benefits. The mass of sucrose needed to produce 1.0 g curdlan by Agrobacterium sp. ATCC 31749 in CK is 1.7-fold more than in 10% MJDA. CONCLUSION: The results strongly revealed that 5-10% MJDA is a good curdlan fermentation media which increase curdlan production yield with cheaper cost of production and simultaneously reduce environmental waste resulting from the large scaled discarded bottom parts of green Asparagus spear during Asparagus production.