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1.
Food Chem ; 334: 127428, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32688173

RESUMO

Aspergillus quadrilineatus endo-ß-mannanase effectively degraded konjac glucomannan (66.09% w/v), copra meal (38.99% w/v) and locust bean galactomannan (20.94% w/v). High performance liquid chromatography (HPLC) analysis of KG hydrolysate indicated its mannooligosaccharides (MOS) content (656.38 mg/g) with high amounts of DP 5 oligosaccharide. Multi-scale characterization of mannan hydrolysate was done using FTIR and 13C NMR which revealed α and ß form of galactose or glucose in MOS, respectively. CM and LBG hydrolysates (1 mg/mL) have shown cytotoxic effect and reduced cell viability of Caco-2 cells by 45% and 62%, respectively. MOS DP (1-4) derived from LBG supported better Lactobacilli biofilm formation as compared to KG hydrolysate containing high DP MOS (5-7). Lactobacilli effectively fermented MOS to generate acetate and propionate as main short chain fatty acids. Lactobacilli produced leucine, isoleucine and valine as branched chain amino acids when grown on LBG hydrolysate.


Assuntos
Mananas/química , Oligossacarídeos/farmacologia , Prebióticos , beta-Manosidase/metabolismo , Aspergillus/enzimologia , Biofilmes , Células CACO-2 , Fermentação , Humanos , Hidrólise , Lactobacillus/crescimento & desenvolvimento , Espectroscopia de Ressonância Magnética , Mananas/metabolismo , Oligossacarídeos/química , Oligossacarídeos/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , beta-Manosidase/química
2.
PLoS One ; 15(9): e0238381, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32881942

RESUMO

Small fructans improve plant tolerance for cold stress. However, the underlying molecular mechanisms are poorly understood. Here, we have demonstrated that the small fructan tetrasaccharide nystose improves the cold stress tolerance of primary rice roots. Roots developed from seeds soaked in nystose showed lower browning rate, higher root activity, and faster growth compared to seeds soaked in water under chilling stress. Comparative proteomics analysis of nystose-treated and control roots identified a total of 497 differentially expressed proteins. GO classification and KEGG pathway analysis documented that some of the upregulated differentially expressed proteins were implicated in the regulation of serine/threonine protein phosphatase activity, abscisic acid-activated signaling, removal of superoxide radicals, and the response to oxidative stress and defense responses. Western blot analysis indicated that nystose promotes the growth of primary rice roots by increasing the level of RSOsPR10, and the cold stress-induced change in RSOsPR10levelis regulated by jasmonate, salicylic acid, and abscisic acid signaling pathways in rice roots. Furthermore, OsMKK4-dependentmitogen-activated protein kinase signaling cascades may be involved in the nystose-induced cold tolerance of primary rice roots. Together, these results indicate that nystose acts as an immunostimulator of the response to cold stress by multiple signaling pathways.


Assuntos
Resposta ao Choque Frio/efeitos dos fármacos , Oligossacarídeos/farmacologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteômica/métodos , Transdução de Sinais/efeitos dos fármacos , Ácido Abscísico/metabolismo , Cromatografia Líquida de Alta Pressão , Resposta ao Choque Frio/genética , Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oryza/efeitos dos fármacos , Oryza/crescimento & desenvolvimento , Oxilipinas/metabolismo , Fenótipo , Proteínas de Plantas/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Ácido Salicílico/metabolismo , Transdução de Sinais/genética , Espectrometria de Massas em Tandem
3.
PLoS One ; 15(8): e0238006, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32857814

RESUMO

This study aimed to evaluate the effects of two prebiotics in different concentrations on nutrient digestibility, fermentative products and immunological variables in adult dogs. Twenty-four adult dogs were randomly divided into six blocks according to their metabolic body weights (BW0.75); within these groups, dogs were randomized to four treatments: control without prebiotics (CO); inclusion of 0.5% prebiotic blend Yes-Golf (B1); inclusion of 1.0% galactooligosaccharide (GOS); and inclusion of 1.0% prebiotic blend Yes-Golf (B2). The experiment lasted 30 days, with 20 days adaptation and 10 days stool and blood collection. Results were analyzed for normality and means were separated by ANOVA and adjusted by the Tukey test at the significance level of 5.0%. Prebiotic supplementation had no effect on apparent digestibility coefficients (ADC), total stool production and fecal scores (p > 0.05). Prebiotics evaluated also did not alter fecal pH, nor the concentrations of ammonia, lactic acid, short chain fatty acids (SCFA) and most fecal branched chain fatty acids (BCFA) (p > 0.05). The addition of GOS decreased the concentration of iso-valeric acid (p = 0.0423). Regarding immunological variables, concentrations of fecal IgA were not influenced by the treatments. Treatments GOS and B2 increased the total number of polymorphonuclear cells, as well as the oxidative burst in relation to treatments B1 and CO (p < 0.0001). Treatment B2 improved the rate of S. aureus phagocytosis in relation to CO (p = 0.0111), and both the GOS and B2 treatments had a better index for E. coli phagocytosis than the CO treatment (p = 0.0067). In conclusion, there was indication that both prebiotics GOS and B2 at 1.0% inclusion improved the immunity of healthy dogs.


Assuntos
Colo/efeitos dos fármacos , Oligossacarídeos/farmacologia , Prebióticos , Animais , Colo/imunologia , Colo/microbiologia , Dieta/veterinária , Cães , Ácidos Graxos Voláteis/análise , Fezes/química , Microbioma Gastrointestinal/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Leucócitos/citologia , Leucócitos/efeitos dos fármacos , Leucócitos/metabolismo , Fagocitose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/fisiologia
4.
Life Sci ; 258: 118085, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32663578

RESUMO

BACKGROUND: An integral intestinal barrier is essential for intestinal homeostasis. Yet, as a side effect of cancer treatment, chemotherapeutic drugs have been reported to cause mucositis. In a recent study, we found that alginate oligosaccharides (AOS) prevent busulfan induced intestinal mucositis. However, it is not known if AOS improves small intestine epithelial cell integrity and migration, which are two essential processes for maintaining the mechanical barrier function of the small intestine. In the current investigation, we aimed to explore the effects of AOS on the integrity and migration of small intestine cells using swine intestinal epithelial IPEC-J2 cells. METHODS: Cell integrity was determined using the TEER assay. Cell migration capability was detected using a wound healing experiment. Small interfering RNA (siRNA) was used to inhibit mannose receptor (MR) expression. Western blotting and immunofluorescence staining were used to determine protein expression. RESULTS: Increasing levels of AOS improved cell integrity as measure by TEER. At the same time, AOS improved IPEC-J2 cell migration capacity as shown in the wound closure assay. It is interesting to note that AOS increased the expression of intestinal microvillus proteins and junction proteins to benefit cell integrity. MR siRNA blocked the action of AOS on cell integrity and cell migration and inhibited the expression of microvillus and cell junction proteins. CONCLUSION: We identified the underlying mechanisms by which AOS improved small intestinal mucositis. As a novel, natural food additive, AOS may be administered to prevent intestinal mucositis induced by chemotherapy or other issues.


Assuntos
Alginatos/farmacologia , Movimento Celular/efeitos dos fármacos , Intestino Delgado/citologia , Oligossacarídeos/farmacologia , Animais , Linhagem Celular , Lectinas Tipo C/metabolismo , Lectinas de Ligação a Manose/metabolismo , Proteínas dos Microfilamentos/metabolismo , Microvilosidades/efeitos dos fármacos , Microvilosidades/metabolismo , Miosinas/metabolismo , RNA Interferente Pequeno/metabolismo , Receptores de Superfície Celular/metabolismo , Suínos , Proteínas de Junções Íntimas/metabolismo , Cicatrização/efeitos dos fármacos
5.
J Nutr ; 150(8): 2077-2088, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32542361

RESUMO

BACKGROUND: The intestinal epithelial cells, food molecules, and gut microbiota are continuously exposed to intestinal peristaltic shear force. Shear force may impact the crosstalk of human milk oligosaccharides (hMOs) with commensal bacteria and intestinal epithelial cells. OBJECTIVES: We investigated how hMOs combined with intestinal peristaltic shear force impact intestinal epithelial cells and crosstalk with a commensal bacterium. METHODS: We applied the Ibidi system to mimic intestinal peristaltic shear force. Caco-2 cells were exposed to a shear force (5 dynes/cm2) for 3 d, and then stimulated with the hMOs, 2'-fucosyllactose (2'-FL), 3-FL, and lacto-N-triose II (LNT2). In separate experiments, Lactobacillus plantarumWCFS1 adhesion to Caco-2 cells was studied with the same hMOs and shear force. Effects were tested on gene expression of glycocalyx-related molecules (glypican 1 [GPC1], hyaluronan synthase 1 [HAS1], HAS2, HAS3, exostosin glycosyltransferase 1 [EXT1], EXT2), defensin ß-1 (DEFB1), and tight junction (tight junction protein 1 [TJP1], claudin 3 [CLDN3]) in Caco-2 cells. Protein expression of tight junctions was also quantified. RESULTS: Shear force dramatically decreased gene expression of the main enzymes for making glycosaminoglycan side chains (HAS3 by 43.3% and EXT1 by 68.7%) (P <0.01), but did not affect GPC1 which is the gene responsible for the synthesis of glypican 1 which is a major protein backbone of glycocalyx. Expression of DEFB1, TJP1, and CLDN3 genes was decreased 60.0-94.9% by shear force (P <0.001). The presence of L. plantarumWCFS1 increased GPC1, HAS2, HAS3, and ZO-1 expression by 1.78- to 3.34-fold (P <0.05). Under shear force, all hMOs significantly stimulated DEFB1 and ZO-1, whereas only 3-FL and LNT2 enhanced L. plantarumWCFS1 adhesion by 1.85- to 1.90-fold (P <0.01). CONCLUSIONS: 3-FL and LNT2 support the crosstalk between the commensal bacterium L. plantarumWCFS1 and Caco-2 intestinal epithelial cells, and shear force can increase the modulating effects of hMOs.


Assuntos
Células Epiteliais/efeitos dos fármacos , Mucosa Intestinal/citologia , Lactobacillus plantarum/efeitos dos fármacos , Leite Humano/química , Oligossacarídeos/farmacologia , Células CACO-2 , Células Epiteliais/fisiologia , Humanos , Lactobacillus plantarum/fisiologia , Peristaltismo
6.
J Anim Sci ; 98(6)2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32497185

RESUMO

The outer cell wall of yeast is characterized by high levels of ß-glucans and mannan-oligosaccharides (MOS), which have been linked with beneficial effects on intestinal health and immune status in dogs. In this study, a standardized in vitro simulation of the canine gastrointestinal tract (Simulator of the Canine Intestinal Microbial Ecosystem; SCIME) was used to evaluate the effect of a Saccharomyces cerevisiae-based product, consisting of 27.5% ß-glucans and 22.5% MOS, on the activity (as assessed by measurement of fermentative metabolites) and composition (as assessed by 16S-targeted Illumina sequencing) of canine intestinal microbiota. The S. cerevisiae-based product was tested at three different dosages, i.e., 0.5, 1.0, and 2.0 g/d. A dose-dependent fermentation pattern was observed along the entire length of the colon, as shown by the increased production of the health-related acetate, propionate, and butyrate for the three concentrations tested (0.5, 1.0, and 2.0 g/d). A consistent finding for all three tested concentrations was the increased propionate production (P < 0.05) in the simulated proximal and distal colon. These changes in terms of fermentative metabolites could be linked to specific microbial alterations at the family level, such as the specific stimulation of the propionate-producing families Porphyromonadaceae and Prevotellaceae upon in vitro exposure to the S. cerevisiae-based product. Other consistent changes in community composition upon repeated exposure included the decrease in the Enterobacteriaceae and the Fusobacteriaceae families, which both contain several potentially opportunistic pathogens. Altogether, the generated data support a possible health-promoting role of a product high in ß-glucans and MOS when supplemented to the dogs' diet.


Assuntos
Suplementos Nutricionais/análise , Cães/fisiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Mananas/farmacologia , Oligossacarídeos/farmacologia , Saccharomyces cerevisiae/química , beta-Glucanas/farmacologia , Animais , Parede Celular/química , Dieta/veterinária , Cães/microbiologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/crescimento & desenvolvimento , Fermentação , Fusobactérias/efeitos dos fármacos , Fusobactérias/crescimento & desenvolvimento , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Fermento Seco/química
7.
Food Chem ; 329: 127179, 2020 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-32505987

RESUMO

The prebiotic activities of hydrolyzed guar gum (GMOS, <1 kDa; GMPS, 1-10 kDa), manno-oligosaccharide (MOS, <1 kDa), and galacto-oligosaccharide (GOS, <1 kDa) were evaluated by in vitro fermentation. The tested carbohydrates showed selective prebiotic effects on bacterial growth, short-chain fatty acid (SCFA)-production, and substrate consumption. GOS and GMOS markedly promoted the growth of bifidobacteria and Clostridium butyricum, respectively, whereas MOS showed the strongest butyrogenic effect. Moreover, SCFA production in the hydrolyzed guar gum groups was closely related to the varied molecular weight (Mw) of the hydrolysate. During in vitro fermentation with human fecal inocula, GMOS gave the highest yields of lactate, propionate, and butyrate after 48 h fermentation. Combined application of MOS and C. butyricum increased the abundance of Clostridiaceae_1. Overall, our results indicate that galactosyl and mannosyl carbohydrates have individualized prebiotic effects which are associated with their chemical structures including their glycoside composition and Mw.


Assuntos
Oligossacarídeos/análise , Prebióticos/análise , Técnicas de Cultura Celular por Lotes , Bifidobacterium/efeitos dos fármacos , Bifidobacterium/genética , Bifidobacterium/crescimento & desenvolvimento , Cromatografia Líquida de Alta Pressão , Clostridium butyricum/efeitos dos fármacos , Clostridium butyricum/genética , Clostridium butyricum/crescimento & desenvolvimento , Ácidos Graxos Voláteis/química , Ácidos Graxos Voláteis/metabolismo , Fezes/microbiologia , Galactanos/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Mananas/metabolismo , Oligossacarídeos/química , Oligossacarídeos/farmacologia , Gomas Vegetais/metabolismo , RNA Ribossômico 16S/química , RNA Ribossômico 16S/metabolismo
8.
J Dairy Sci ; 103(7): 5816-5829, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32418689

RESUMO

Fermented milk is an effective carrier for probiotics, the consumption of which improves host health. The beneficial effects of probiotics, prebiotics, and synbiotics on gut dysbiosis have been reported previously. However, the way in which specific probiotics, prebiotics, and synbiotics regulate intestinal microbes remains unclear. Therefore, the probiotics Lactobacillus rhamnosus AS 1.2466 and Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 and the prebiotics xylooligosaccharide and red ginseng extracts were fed to mice to determine their effects on the intestinal microbiota. Then, mice were administered xylooligosaccharide and L. rhamnosus (synthesis) by gavage, and the number of L. rhamnosus was determined in the intestine at different times. The results show that probiotics and prebiotics can quickly reduce the Firmicutes/Bacteroidetes ratio, inhibit harmful bacteria (such as Klebsiella and Escherichia coli), and accelerate the recovery of beneficial intestinal microorganisms (such as Lactobacillus). In a complex intestinal microecology, different probiotics and prebiotics have different effects on specific intestinal microorganisms that cannot be recovered in the short term. In addition, after 20 d of intragastric xylooligosaccharide addition at 0.12 g/kg of body weight, L. rhamnosus colonization in the mouse ileum was 7.48 log cfu/mL, which was higher than in the low-dose group, prolonging colonization time and increasing the number of probiotics in the intestine. Therefore, this study demonstrated that probiotics and prebiotics can promote the balance of intestinal microbiota by regulating specific microbes in the intestine, and the effects of a suitable combination of synbiotics are beneficial, laying the foundation for the development of new dairy products rich in synbiotics.


Assuntos
Bactérias/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Prebióticos , Probióticos/farmacologia , Simbióticos , Ampicilina/farmacologia , Animais , Antibacterianos/farmacologia , Microbioma Gastrointestinal/fisiologia , Glucuronatos/administração & dosagem , Glucuronatos/farmacologia , Lactobacillus delbrueckii/química , Lactobacillus rhamnosus/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oligossacarídeos/administração & dosagem , Oligossacarídeos/farmacologia , Panax/química , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Prebióticos/administração & dosagem , Probióticos/administração & dosagem , Organismos Livres de Patógenos Específicos , Simbióticos/administração & dosagem
9.
J Biotechnol ; 318: 31-38, 2020 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-32387450

RESUMO

Norovirus infections cause severe gastroenteritis in millions of people every year. Infection requires the recognition of histo-blood group antigens (HBGAs), but such interactions can be inhibited by human milk oligosaccharides (HMOs), which act as structurally-similar decoys. HMO supplements could help to prevent norovirus infections, but the industrial production of complex HMOs is challenging. Here we describe a large-scale fermentation process that yields several kilograms of lacto-N-fucopentaose I (LNFP I). The product was synthesized in Escherichia coli BL21(DE3) cells expressing a recombinant N-acetylglucosaminyltransferase, ß(1,3)galactosyltransferase and α(1,2)fucosyltransferase. Subsequent in vitro enzymatic conversion produced HBGA types A1 and B1 for norovirus inhibition assays. These carbohydrates inhibited the binding of GII.17 virus-like particles (VLPs) to type A1 and B1 trisaccharides more efficiently than simpler fucosylated HMOs, which were in turn more effective than any non-fucosylated structures. However, we found that the simpler fucosylated HMOs were more effective than complex molecules such as LNFP I when inhibiting the binding of GII.17 and GII.4 VLPs to human gastric mucins and mucins from human amniotic fluid. Our results show that complex fucosylated HMOs can be produced by large-scale fermentation and that a combination of simple and complex fucosylated structures is more likely to prevent norovirus infections.


Assuntos
Norovirus/efeitos dos fármacos , Oligossacarídeos/metabolismo , Oligossacarídeos/farmacologia , Receptores Virais/metabolismo , Biotecnologia , Antígenos de Grupos Sanguíneos/química , Antígenos de Grupos Sanguíneos/metabolismo , Antígenos de Grupos Sanguíneos/farmacologia , Fermentação , Humanos , Concentração Inibidora 50 , Leite Humano/química , Mucinas/metabolismo , Norovirus/fisiologia , Oligossacarídeos/química , Trissacarídeos/metabolismo
10.
Food Microbiol ; 90: 103494, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32336371

RESUMO

P. psychrophila is implicated in fish spoilage especially under cold storage. In the present study, tandem mass tag (TMT) quantitative proteomic analysis was performed to clarify the molecular mechanism for the inhibitory effect of chitosan oligosaccharides (COS) against P. psychrophila in fish juice system. The MIC and MBC of the COS against P. psychrophila were 6 and 8 mg/mL, respectively. Compared with the untreated control, a total of 370 proteins (163 up-regulated and 207 down-regulated) were identified as differentially expressed proteins (DEPs, >1.5-fold or < 0.67-fold, P < 0.05) in P. psychrophila when exposed to 6 mg/mL COS. Bioinformatics analysis indicated that the DEPs were mainly involved in the cell wall/membrane, cell motility, and electron-transport chain; DNA replication, RNA transcription and translation, posttranslational modifications of proteins; TCA cycle, and the transport and metabolism of amino acid, carbohydrate, and ion. The scanning electron microscope (SEM) and fourier-transform infrared spectroscopy (FT-IR) analysis further validated that cell structure especially the cell wall/membrane was damaged after COS treatment. The results in this study presented an important step toward understanding the response of P. psychrophila cells to COS at the proteome level.


Assuntos
Quitosana/farmacologia , Peixes/microbiologia , Oligossacarídeos/farmacologia , Proteômica/métodos , Pseudomonas/genética , Animais , Membrana Celular/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Oligossacarídeos/química , Biossíntese de Proteínas/efeitos dos fármacos , Pseudomonas/química , Pseudomonas/efeitos dos fármacos , Transcrição Genética/efeitos dos fármacos
11.
Adv Exp Med Biol ; 1221: 473-491, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32274723

RESUMO

The heparan sulfate mimetic PI-88 (muparfostat) is a complex mixture of sulfated oligosaccharides that was identified in the late 1990s as a potent inhibitor of heparanase. In preclinical animal models it was shown to block angiogenesis, metastasis and tumor growth, and subsequently became the first heparanase inhibitor to enter clinical trials for cancer. It progressed to Phase III trials but ultimately was not approved for use. Herein we summarize the preparation, physicochemical and biological properties of PI-88, and discuss preclinical/clinical and structure-activity relationship studies. In addition, we discuss the PI-88-inspired development of related HS mimetic heparanase inhibitors with improved properties, ultimately leading to the discovery of PG545 (pixatimod) which is currently in clinical trials.


Assuntos
Antineoplásicos/farmacologia , Heparitina Sulfato/farmacologia , Neoplasias/tratamento farmacológico , Oligossacarídeos/farmacologia , Animais , Antineoplásicos/uso terapêutico , Glucuronidase/antagonistas & inibidores , Heparitina Sulfato/química , Humanos , Neoplasias/irrigação sanguínea , Neoplasias/enzimologia , Neovascularização Patológica/tratamento farmacológico , Oligossacarídeos/química , Oligossacarídeos/uso terapêutico , Relação Estrutura-Atividade
12.
J Pharmacol Sci ; 143(2): 65-73, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32229084

RESUMO

Glucagon-like peptide 1 (GLP-1) released from enteroendocrine (L) cells regulates insulin secretion. Intestinal inflammation and impaired GLP-1 release have been found in type 2 diabetes mellitus (T2DM) patients. Fructo-oligosaccharides (FOS), a known prebiotic, improve GLP-1 release and glucose homeostasis in T2DM models. This study aimed to investigate the effect of tumor necrosis factor-α (TNF-α), a proinflammatory cytokine associated with intestinal inflammation in T2DM, on L cell apoptosis and the effect of FOS on inflammation-associated impairment of GLP-1 secretion. Herein, using cell death assays, immunofluorescence staining, real time PCR and Western blot analyses, we found that TNF-α induced L cell apoptosis via nuclear factor kappa B (NF-κB)- inducible nitric oxide synthase (iNOS)-cleaved caspase-3-dependent pathways. Interestingly, FOS did not suppress TNF-α-induced NF-κB nuclear translocation, but inhibited expression of iNOS and cleaved caspase-3. In addition, FOS alleviated apoptosis and rescued impaired GLP-1 release in TNF-α-treated L cells. Altogether, our data indicate that TNF-α induces L cell apoptosis via an NF-κB-iNOS-caspase-3-dependent pathway. FOS may be useful in suppressing inflammation-associated L cell apoptosis and maintaining GLP-1 level in T2DM patients.


Assuntos
Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Células Enteroendócrinas/metabolismo , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Oligossacarídeos/farmacologia , Apoptose/genética , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Inflamação , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo
13.
Mar Drugs ; 18(2)2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-32023889

RESUMO

Alginate extracted from widely cultured brown seaweed can be hydrolyzed by alginate lyase to produce alginate oligosaccharides (AOS) with intriguing biological activities. Herein, a novel alginate lyase Aly1281 was cloned from marine bacterium Pseudoalteromonas carrageenovora ASY5 isolated from mangrove soil and found to belong to polysaccharide lyase family 7. Aly1281 exhibited maximum activity at pH 8.0 and 50 °C and have broad substrate specificity for polyguluronate and polymannuronate. Compared with other alginate lyases, Aly1281 exhibited high degradation specificity and mainly produced di-alginate oligosaccharides which displayed good antioxidant function to reduce ferric and scavenge radicals such as hydroxyl, ABTS+ and DPPH. Moreover, the catalytic activity and kinetic performance of Aly1281 were highly improved with the addition of salt, demonstrating a salt-activation property. A putative conformational structural feature of Aly1281 was found by MD simulation analysis for understanding the salt-activation effect.


Assuntos
Polissacarídeo-Liase/isolamento & purificação , Pseudoalteromonas/enzimologia , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Concentração de Íons de Hidrogênio , Oligossacarídeos/metabolismo , Oligossacarídeos/farmacologia , Polissacarídeo-Liase/química , Polissacarídeo-Liase/metabolismo , Pseudoalteromonas/isolamento & purificação , Microbiologia do Solo , Especificidade por Substrato , Temperatura
14.
PLoS One ; 15(2): e0229281, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32092087

RESUMO

The aim of this study was to investigate the effect on growth performance and nutrient utilisation when supplementing diets deficient in energy and protein with carbohydrase enzymes or xylo-oligosaccharide in broilers challenged with coccidia. 960 Ross 308 broilers were used in this 21-day study. The treatments were arranged into a 2×4 factorial with 2 challenge states (challenged and non-challenged) and 4 different additive types (control, xylanase alone, xylanase and ß-glucanase mixture and xylo-oligosaccharide). On day 14, the challenged group received 12× the recommended dose of coccidiosis vaccine while the non-challenged group received a sham treatment of water only. The birds and feed were weighed on days 0, 14 and 21. On day 21, two birds per pen were euthanized, the caeca were removed and the contents collected for short chain fatty acid analysis. Six more birds per pen were euthanized and ileal digesta were collected and pooled per pen for nutrient digestibility analysis. Feed intake was greater (P < 0.05) on days 14 and 21 when xylo-oligosaccharide was included in the diet compared to the xylanase and ß-glucanase mixture in birds challenged with coccidiosis. Including xylo-oligosaccharide in the diet improved (P < 0.05) the digestibility of nitrogen and supplementing diets with the xylanase and ß-glucanase mixture improved (P < 0.05) the digestibility of several amino acids. The concentration of arabinose and xylose was (P < 0.001) greater when broiler diets were supplemented with carbohydrase enzymes or xylo-oligosaccharide compared to the control. Although there was an increase in short chain fatty acid production due to the addition of carbohydrase enzymes or xylo-oligosaccharide, there was no additive effect on the %G+C profile of caecal bacteria however there was a negative effect of coccidiosis. In conclusion, the similarity in the response to carbohydrase enzymes or xylo-oligosaccharide supplementation illustrates that the hydrolysis products from carbohydrase activity may have prebiotic like effects.


Assuntos
Galinhas/crescimento & desenvolvimento , Coccidiose , Glicosídeo Hidrolases/farmacologia , Prebióticos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Galinhas/parasitologia , Coccidiose/dietoterapia , Coccidiose/tratamento farmacológico , Dieta , Suplementos Nutricionais , Digestão , Grão Comestível , Ácidos Graxos Voláteis/análise , Glicosídeo Hidrolases/metabolismo , Oligossacarídeos/farmacologia
15.
Poult Sci ; 99(2): 1135-1149, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32036965

RESUMO

Salmonella and Campylobacter are considered major public health burdens worldwide, and poultry are known to be one of the main reservoirs for these zoonotic pathogens. This study was conducted to evaluate the effect of a commercial probiotic or direct-fed microbial (DFM) Calsporin (CSP), and prebiotic or mannan oligosaccharide (MOS) (IMW50) on ultrastructural changes and the villous integrity of intestinal mucosa in turkey poults challenged with Salmonella and Campylobacter. A 21-day battery cage study was conducted using 4 dietary treatments including a basal diet (corn and soybean-based) nonsupplemented and uninfected as a negative control (NC); basal diet supplemented with 0.05% DFM (CSP); basal diet supplemented with 0.05% MOS (IMW50); and basal diet supplemented with 0.05% mixture of DFM and MOS at equal proportions. Female large white turkey poults aged 336 days were obtained from a local commercial hatchery and randomly distributed in electrically heated battery cages with 12 treatments of 4 replicates per treatment containing 7 poults per pen. The first 16 pens were not infected with bacteria, poults in pens 17-32 were orally challenged at day 7 with 105 cfu Salmonella Heidelberg, and the poults in pens 33-48 were orally challenged at day 7 with 105 cfu Campylobacter jejuni. Feed and water were provided ad libitum throughout the study. At day 21, ileal tissue samples from 1 bird per cage were collected for intestinal integrity and ultrastructural examination by scanning and electron microscopy. DFM and MOS supplementation was effective in both challenged and nonchallenged (not infected with Salmonella and Campylobacter) birds. Goblet cells and mucus were increased, with the presence of large numbers of segmented filamentous bacteria in DFM- and MOS-supplemented groups compared with birds in control treatments. The number and size of villi were reduced in poults exposed to Salmonella and Campylobacter. Results show that CSP and IMW50 provide protection of ileal mucosal integrity in poults exposed to Salmonella or Campylobacter.


Assuntos
Infecções por Campylobacter/veterinária , Doenças das Aves Domésticas/prevenção & controle , Prebióticos , Probióticos/farmacologia , Salmonelose Animal/prevenção & controle , Perus , Ração Animal/análise , Animais , Campylobacter/fisiologia , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/prevenção & controle , Dieta/veterinária , Suplementos Nutricionais/análise , Íleo/efeitos dos fármacos , Íleo/ultraestrutura , Mucosa Intestinal/efeitos dos fármacos , Mananas/farmacologia , Microscopia Eletrônica de Varredura/veterinária , Microscopia Eletrônica de Transmissão/veterinária , Oligossacarídeos/farmacologia , Distribuição Aleatória , Saccharomyces cerevisiae/química , Salmonella/fisiologia , Salmonelose Animal/microbiologia
16.
Carbohydr Polym ; 234: 115903, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32070523

RESUMO

In this study, 2-urea-chitosan oligosaccharide derivatives (2-urea-COS derivatives) and 2,6-diurea-chitosan oligosaccharide derivatives (2,6-diurea-COS derivatives) were successfully designed and synthesized via intermediate 2-methoxyformylated chitosan oligosaccharide. All samples were characterized and compared based on FT-IR, 1H NMR spectroscopy, and elemental analysis. The antifungal effects of COS derivatives were tested against Fusarium oxysporum f. sp. niveum, Phomopsis asparagus, and Botrytis cinereal. Their antioxidant properties, including superoxide radicals' scavenging activity, hydroxyl radicals' scavenging activity, and DPPH radicals' scavenging activity were also explored within different concentrations. COS derivatives bearing urea groups showed improved bioactivity compared with pristine COS and 2,6-diurea-COS derivatives had a higher biological activity than 2-urea-COS derivatives in tested concentrations. Additionally, L929 cells were used to carry out cytotoxicity test of COS and COS derivatives by CCK-8 assay. The results indicated that some of samples showed low cytotoxicity. These findings offered a suggestion that COS derivatives bearing urea groups are promising biological materials.


Assuntos
Antifúngicos/farmacologia , Antioxidantes/farmacologia , Quitosana/farmacologia , Oligossacarídeos/farmacologia , Antifúngicos/síntese química , Antifúngicos/química , Antioxidantes/síntese química , Antioxidantes/química , Ascomicetos/efeitos dos fármacos , Compostos de Bifenilo/antagonistas & inibidores , Botrytis/efeitos dos fármacos , Quitosana/química , Fusarium/efeitos dos fármacos , Radical Hidroxila/antagonistas & inibidores , Testes de Sensibilidade Microbiana , Estrutura Molecular , Oligossacarídeos/síntese química , Oligossacarídeos/química , Picratos/antagonistas & inibidores , Superóxidos/antagonistas & inibidores
17.
Benef Microbes ; 11(1): 19-32, 2020 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-32066258

RESUMO

Previously, we showed enhanced efficacy of oral immunotherapy (OIT) using fructo-oligosaccharides (FOS, prebiotics) added to the diet of cow's milk allergic mice indicated by a reduction in clinical symptoms and mast cell degranulation. Prebiotics are fermented by gut bacteria, affecting both bacterial composition and availability of metabolites (i.e. short-chain fatty acids (SCFA)). It is thus far unknown which microbial alterations are involved in successful outcomes of OIT with prebiotic supplementation for the treatment of food allergy. To explore potential changes in the microbiota composition and availability of SCFA induced by OIT+FOS. C3H/HeOuJ mice were sensitised and received OIT with or without a FOS supplemented diet. After three weeks, faecal samples were collected to analyse gut microbiota composition using 16S rRNA sequencing. SCFA concentrations were determined in cecum content. FOS supplementation in sensitised mice changed the overall microbial community structure in faecal samples compared to sensitised mice fed the control diet (P=0.03). In contrast, a high level of resemblance in bacterial community structure was observed between the non-sensitised control mice and the OIT+FOS treated mice. OIT mice showed an increased relative abundance of the dysbiosis-associated phylum Proteobacteria compared to the OIT+FOS mice. FOS supplementation increased the relative abundance of genus Allobaculum (Firmicutes), putative butyrate-producing bacteria. OIT+FOS reduced the abundances of the genera's unclassified Rikenellaceae (Bacteroidetes, putative pro-inflammatory bacteria) and unclassified Clostridiales (Firmicutes) compared to sensitised controls and increased the abundance of Lactobacillus (Firmicutes, putative beneficial bacteria) compared to FOS. OIT+FOS mice had increased butyric acid and propionic acid concentrations. OIT+FOS induced a microbial profile closely linked to non-allergic mice and increased concentrations of butyric acid and propionic acid. Future research should confirm whether there is a causal relationship between microbial modulation and the reduction in acute allergic symptoms induced by OIT+FOS.


Assuntos
Hipersensibilidade Alimentar , Oligossacarídeos , Prebióticos/administração & dosagem , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Butiratos/metabolismo , Ceco/metabolismo , Ceco/microbiologia , Dietoterapia/métodos , Ácidos Graxos Voláteis/metabolismo , Fezes/microbiologia , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/microbiologia , Hipersensibilidade Alimentar/terapia , Microbioma Gastrointestinal/efeitos dos fármacos , Imunoterapia/métodos , Lactobacillus/isolamento & purificação , Camundongos , Camundongos Endogâmicos C3H , Microbiota/efeitos dos fármacos , Leite/efeitos adversos , Leite/metabolismo , Oligossacarídeos/administração & dosagem , Oligossacarídeos/farmacologia , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética
18.
Carbohydr Polym ; 233: 115844, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32059896

RESUMO

A glycosaminoglycan was isolated from the sea cucumber Holothuria coluber (HcFG). A series of oligosaccharide fragments (dp range 3-11) were prepared from its ß-eliminative depolymerized product (dHcFG). Extensive NMR characterization of the oligosaccharides indicated the d-GlcA-ß1,3-d-GalNAc4S6S repeating disaccharide backbone was substituted by monosaccharide branches comprising of Fuc2S4S, Fuc3S4S and Fuc4S, linked to O-3 of d-GlcA. For the prevailing Fuc3S4S at nonreducing end of dHcFG, the ß-eliminative depolymerization process of HcFG was compared with those of the FGs from Actinopyga miliaris (AmFG, branched with Fuc3S4S) and Stichopus variegatus (SvFG, branched with Fuc2S4S). The result suggested that d-GlcA substituted with Fuc3S4S was more susceptible to depolymerization than that with Fuc2S4S. It might be due to the larger steric hindrance effects from Fuc2S4S on the esterification of GlcA. Biological assays confirmed that the minimum chain length (dp8), regardless of the Fuc branch types, was required for the potent anti-iXase and anticoagulant activities in FG fragments.


Assuntos
Anticoagulantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Glicosaminoglicanos/farmacologia , Holothuria/química , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Sequência de Carboidratos , Fucose/química , Glicosaminoglicanos/química , Glicosaminoglicanos/isolamento & purificação , Humanos , Peso Molecular , Ressonância Magnética Nuclear Biomolecular , Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , Oligossacarídeos/farmacologia , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Tempo de Trombina
19.
Sci Rep ; 10(1): 768, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31964943

RESUMO

Current treatment options for influenza virus infections in humans are limited and therefore the development of novel antivirals is of high priority. Inhibiting influenza virus attachment to host cells would provide an early and efficient block of the infection and thus, receptor analogs have been considered as options for antiviral treatment. Here, we describe the rapid and efficient synthesis of PAMAM dendrimers conjugated with either 3'-sialyllactose (3SL) or 6'-sialyllactose (6SL) and their potential to inhibit a diverse range of human and avian influenza virus strains. We show in a hemagglutination inhibition (HAI) assay that human IAV strains can be inhibited by (6SL)- and to a lesser extent also by (3SL)-conjugated PAMAM dendrimers. In contrast, avian strains could only be inhibited by (3SL)-conjugated dendrimers. Importantly, the differential sensitivities of human and avian IAV to the two types of sialyllactose-conjugated dendrimers could be confirmed in cell-based neutralization assays. Based on our findings, we suggest to further develop both, (3SL)- and (6SL)-conjugated PAMAM dendrimers, as influenza virus inhibitors.


Assuntos
Antivirais/síntese química , Dendrímeros/química , Vírus da Influenza A/efeitos dos fármacos , Lactose/análogos & derivados , Oligossacarídeos/síntese química , Animais , Antivirais/química , Antivirais/farmacologia , Aves , Embrião de Galinha , Cães , Testes de Inibição da Hemaglutinação , Humanos , Vírus da Influenza A/imunologia , Lactose/síntese química , Lactose/química , Lactose/farmacologia , Células Madin Darby de Rim Canino , Oligossacarídeos/química , Oligossacarídeos/farmacologia , Especificidade da Espécie
20.
Sci Rep ; 10(1): 997, 2020 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-31969618

RESUMO

Lacto-n-neotatraose (LNnT) oligosaccharide shows properties such as anti-inflammatory, type 2 immune response induction, induced angiogenesis, and anti-bacterial effects. Here, we hypothesized that the application of LnNT in the skin full-thickness wound can accelerate the healing process through its anti-inflammatory effect as well as induction of type 2 immune responses. In this study, we evaluated the cell viability of fibroblasts in the presence of LNnT. The full-thickness wound model was created by punch biopsy. The mice were treated intradermaly with LNnT at the concentrations of 100 and 200 µg or PBS as a control group. The wounds samples were compared based on the macroscopic and histological evaluations. The amount of collagen deposition and expression of genes involved in type 2 immunity were measured by the hydroxyproline assay and real time PCR method, respectively. Our results showed that LNnT had no negative effect on the cell viability of fibroblasts. LNnT increased the wound closure rate on day 7 post-wounding. H&E stain analysis revealed that mice treated with 200 µg LNnT exhibited better healing score, follicle formation, and lower epidermal thickness index. The mice treated with LNnT exhibited a lower collagen deposition on day 21 and higher collagen content on days 7 and 14 post-treatment. The LNnT groups also exhibited a lower number of neutrophils and a higher number of basal cells and fibroblasts. The expression rate of IL-10, IL-4, and IL-13 was higher in the LNnT groups. These results showed the high potential of LNnT for use in treatment of full-thickness wounds.


Assuntos
Fibroblastos/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Oligossacarídeos/farmacologia , Pele/efeitos dos fármacos , Cicatrização/genética , Animais , Fibroblastos/metabolismo , Interleucinas/genética , Interleucinas/metabolismo , Camundongos , Pele/metabolismo , Cicatrização/efeitos dos fármacos
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