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Two CO oxidation reactions (CO + O2 â CO2 + O and CO + O â CO2) were considered in the Eley-Rideal (ER) reaction mechanism. These oxidation processes on the W(111) surface and the W helical nanowire were investigated by the density functional theory (DFT) calculation. The stable adsorption sites of O2 and O as well as their adsorption energies were obtained first. In order to understand the catalytic properties of the W helical nanowire, the Fukui function and local density of state (LDOS) profiles were determined. The nudged elastic band (NEB) method was applied to locate transition states and minimum energy pathways (MEPs) of CO oxidation processes on the W helical nanowire and on the W(111) surface. In this study, we have demonstrated that the catalytic ability of the W helical nanowire is superior to that of the W(111) surface for CO oxidation.
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TLR2 recognizes components of Mycobacterium tuberculosis and initiates APC activities that influence both innate and adaptive immunity. M. tuberculosis lipoproteins are an important class of TLR2 ligands. In this study, we focused on recombinant MPT83 (rMPT83) to determine its effects on mouse macrophages. We demonstrated that rMPT83 induced the production of TNF-α, IL-6, and IL-12 p40 and that cytokine induction depended on activated MAPKs, because we observed the rapid phosphorylation of ERK1/2, p38, and JNK in macrophages. Additionally, neutralizing Abs against TLR2 significantly inhibited cytokine secretion and reduced or attenuated the rMPT83-induced activation of p38 and JNK in RAW264.7 cells, a mouse macrophage cell line. Furthermore, rMPT83-induced cytokine production was significantly lower in macrophages from TLR2(-/-) mice than in macrophages from wild-type mice. We further found that prolonged exposure (>24 h) of RAW264.7 cells or macrophages from wild-type and TLR2(-/-) mice to rMPT83 resulted in a significant enhancement of IFN-γ-induced MHC class II expression and an enhanced ability of macrophages to present the rMPT83 peptide to CD4(+) T cells. These results indicated that rMPT83 is a TLR2 agonist that induces the production of cytokines by macrophages and upregulates macrophage function.
Assuntos
Antígenos de Bactérias/fisiologia , Proteínas de Bactérias/fisiologia , Citocinas/biossíntese , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Proteínas de Membrana/fisiologia , Receptor 2 Toll-Like/fisiologia , Regulação para Cima/imunologia , Animais , Antígenos de Bactérias/genética , Proteínas de Bactérias/agonistas , Proteínas de Bactérias/genética , Linhagem Celular , Células HEK293 , Humanos , Subunidade p40 da Interleucina-12/biossíntese , Interleucina-6/biossíntese , Proteínas de Membrana/agonistas , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas Recombinantes/agonistas , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Receptor 2 Toll-Like/deficiência , Fator de Necrose Tumoral alfa/biossínteseRESUMO
This study aimed to identify and quantify the primary components in lotus leaf and to explore the hypolipidemic components through spectral-effect relationships and chemometric methods. Utilizing a data-dependent acquisition-diagnostic fragment ion/characteristic neutral loss screening strategy (DFI-NLS), a reliable HPLC-Q-TOF-MS analysis was conducted, identifying 77 compounds, including 36 flavonoids, 21 alkaloids, 3 terpenoids, 11 organic acids, 4 phenols, 1 lignin and 1 unsaturated hydrocarbon. A straightforward HPLC-DAD method was developed for the simultaneous determination of seven major components in lotus leaf, and quercetin-3-O-glucuronide (Q3GA) was identified as the most abundant component. The HPLC fingerprints of 36 lotus leaf sample batches were assessed using chemometric approaches such as principal component analysis and hierarchical cluster analysis. The hypolipidemic effect of these samples was analyzed by measuring total cholesterol (TC) and total triglycerides (TG) levels in palmitic acid (PA) and oleic acid (OA)-induced lipid modeling in HepG-2 cells, employing partial least squares regression and grey relation analysis to investigate the spectral-effect relationship of the lotus leaf. The in vivo hypolipidemic effect of these compounds was assessed using an egg yolk powder-induced high-fat zebrafish model. The findings indicated that peak No.11 (Q3GA) in the chemical fingerprint was significantly associated with hypolipidemic activity, suggesting it as a potential hypolipidemic compound in lotus leaf. In summary, this study facilitates the exploration of the phytochemical compounds and their bioactive properties in the lotus leaf.
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This study used both the element occurrence form analysis and the chronic health risk assessment method to investigate the accumulation characteristics of heavy metals in the soil-crop system and the health risk assessment of agricultural products in northeastern Yunnan, which is a typical area of Southwest China where heavy metals are enriched in soil. Based on the study of 1137 groups of agricultural products and corresponding root soils, the results showed that the soil cadmium (Cd) and lead (Pb) in the lead-zinc ore concentration area were higher than the risk-intervention values of the "Soil Environmental Quality Risk control standard for soil contamination of agricultural land" (GB 15618-2018), whereas the soil Cd in the other parent material areas was within the screening-intervention buffer zone, and Pb was below the minimum risk-screening value. According to the National Food Safety Standard of China (GB 2762-2017), the heavy metal Cd in potatoes and soybeans in the area seriously exceeded the standard, the heavy metal Pb in tartary buckwheat and walnut exceeded the limit value, and the exceeding rate of heavy metal Cd in crops from these parent material areas showed: clastic rock>basalt>lead-zinc ore>carbonate ≈ Quaternary sedimentary>sand (mud) rock. According to the U.S. Environmental Protection Agency's assessment method for the chronic health risk of heavy metal intake by humans, the grains and potatoes, staple foods, and fruits had low chronic health risks of heavy metal intake. Agricultural products from the parent material area of clastic rock, sand (mud) rock, Quaternary sedimentary, and lead-zinc ore concentration showed health risks; with the change in soil physical and chemical properties and the increase in the types of edible crops, the risk will gradually increase. Based on this research, it is urgent to carry out real-time monitoring of agricultural products in the area.
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Metais Pesados , Poluentes do Solo , Humanos , Solo/química , Cádmio/análise , Areia , Chumbo/análise , China , Poluentes do Solo/análise , Metais Pesados/análise , Zinco , Medição de Risco , Produtos Agrícolas/química , Monitoramento AmbientalRESUMO
BACKGROUND: A recent investigation showed that the prevalence of type 2 diabetes mellitus (T2DM) is 12.8% among individuals of Han ethnicity. Gut microbiota has been reported to play a central role in T2DM. Goto-Kakizaki (GK) rats show differences in gut microbiota compared to non-diabetic rats. Previous studies have indicated that berberine could be successfully used to manage T2DM. We sought to understand its hypoglycaemic effect and role in the regulation of the gut microbiota. AIM: To determine whether berberine can regulate glucose metabolism in GK rats via the gut microbiota. METHODS: GK rats were acclimatized for 1 wk. The GK rats were randomly divided into three groups and administered saline (Mo), metformin (Me), or berberine (Be). The observation time was 8 wk, and weight, fasting blood glucose (FBG), insulin, and glucagon-like peptide-1 (GLP-1) were measured. Pancreatic tissue was observed for pathological changes. Additionally, we sequenced the 16S rRNA V3-V4 region of the gut microbiota and analysed the structure. RESULTS: Compared with the Mo group, the Me and Be groups displayed significant differences in FBG (P < 0.01) and GLP-1 (P < 0.05). A significant decrease in weight and homeostatic model assessment-insulin resistance was noted in the Be group compared with those in the Me group (P < 0.01). The pancreatic islets of the Me- and Be-treated rats showed improvement in number, shape, and necrosis compared with those of Mo-treated rats. A total of 580 operational taxonomic units were obtained in the three groups. Compared to the Mo group, the Me and Be groups showed a shift in the structure of the gut microbiota. Correlation analysis indicated that FBG was strongly positively correlated with Clostridia_UCG-014 (P < 0.01) and negatively correlated with Allobaculum (P < 0.01). Body weight showed a positive correlation with Desulfovibrionaceae (P < 0.01) and a negative correlation with Akkermansia (P < 0.01). Importantly, our results demonstrated that Me and Be could significantly decrease Bacteroidetes (P < 0.01) and the Bacteroidetes/Firmicutes ratio (P < 0.01). Furthermore, Muribaculaceae (P < 0.01; P < 0.05) was significantly decreased in the Me and Be groups, and Allobaculum (P < 0.01) was significantly increased. CONCLUSION: Berberine has a substantial effect in improving metabolic parameters and modulating the gut microbiota composition in T2DM rats.
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Berberina , Diabetes Mellitus Tipo 2 , Microbioma Gastrointestinal , Hiperglicemia , Animais , Berberina/farmacologia , Berberina/uso terapêutico , Glicemia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Hiperglicemia/tratamento farmacológico , RNA Ribossômico 16S/genética , RatosRESUMO
Traditional autotrophic microalgae exhibit low rates of organic carbon assimilation and conversion to useful compounds when switching to mixotrophic or heterotrophic growth. The goal of this study was to investigate the effect of inorganic carbon limitation on the efficiency of organic carbon (glycerol) assimilation and conversion to total fatty acids (TFAs) or the long-chain polyunsaturated fatty acid eicosapentaenoic acid (EPA). An oleaginous Monodus subterraneus was selected and six cultivation conditions were set, including Autotrophy-no aeration, Autotrophy-aeration, Mixotrophy-no aeration, Mixotrophy-no aeration & no Na2CO3, Mixotrophy-aeration, and Heterotrophy. The results showed M. subterraneus could utilize glycerol and grow under mixotrophic condition, while it was not occurred under heterotrophy. Superiority of mixotrophy to autotrophy on biomass productivity was more obvious under inorganic carbon limitation (no aeration or no Na2CO3) than inorganic carbon supply (aeration and existing Na2CO3 in the medium). CO2 limitation (no aeration) decreased content (of dry weight) and production (in medium) of TFAs, which was not evident in mixotrophy. CO2 limitation and inorganic carbon substrate stress largely improved the COD yield of TFAs and EPA under mixotrophic condition. TFA yield (%COD) under Mixotrophy-no aeration & no Na2CO3 was maximum (22.82%), and was almost two-fold higher than that under Mixotrophy-no aeration and nearly three-fold higher than that with Mixotrophy-aeration. EPA yield (% COD) under mixotrophy-no aeration & no Na2CO3 was maximum (6.58%). These results suggested that inorganic carbon limitation is a potentially useful method to enhance conversion of organic carbon to TFAs. Furthermore, the results suggest an application to obtain high value compounds (TFAs or EPA) combined with a high assimilation rate of waste glycerol from biodiesel and epichlorohydrin production by microalgae.
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Microalgas , Estramenópilas , Biomassa , Carbono , Ácidos Graxos , Processos HeterotróficosRESUMO
This study investigated the effect of nitrogen (N) and phosphorous (P) stress on the production of DHA or EPA and total fatty acids (TFAs) in the marine microalga Tisochrysis lutea and the freshwater microalga Monodus subterraneus. Five N or P starvation/limitation conditions (N sufficient and P limited, N sufficient and P starved, N starved and P sufficient, N starved and P limited, and N and P starved) and one N and P sufficient condition (control) were studied. The results demonstrated that the proportion of DHA or EPA among TFAs and production in the microalgae suspensions decreased (57%, 73% for N stress and 18%, 51% for P stress, respectively) under N or P stress in both microalgae compared with the N and P sufficient group. Differently, DHA dry weight content of T. lutea decreased significantly, and EPA dry weight content of M. subterraneus decreased slightly under N starved conditions. Clear differences in TFA content/production and the relationship between TFA and DHA or EPA production/content and CO2 fixation were observed between the two microalgae. These results give a new sight on the difference between marine microalgae and freshwater microalgae. Meanwhile, it gave a potential application to produce DHA or EPA and TFA combining with CO2 fixation by these microalgae.
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Ácidos Docosa-Hexaenoicos/metabolismo , Ácido Eicosapentaenoico/metabolismo , Microalgas/metabolismo , Nitrogênio/metabolismo , Fósforo/metabolismo , Haptófitas/metabolismo , Nitrogênio/deficiência , Nutrientes/deficiência , Nutrientes/metabolismo , Fósforo/deficiência , Estramenópilas/metabolismo , Estresse FisiológicoRESUMO
In this study, Tisochrysis lutea was cultivated in mixotrophic and heterotrophic cultures with glycerol as a carbon source and with glucose and acetate for comparison; autotrophic cultivation was the control group without a carbon source. It was found that T. lutea used glycerol and did not use glucose and acetate under mixotrophy. Mixotrophy slightly elevated the docosahexaenoic acid (DHA) and total fatty acids (TFA) content in the dry-weight and enhanced the DHA and TFA production in medium (41.3 and 31.9% respectively) at the end of a 16-day cultivation, while heterotrophy reduced the DHA content and TFA production. Under the mixotrophy, the glycerol contribution to the DHA production (16.19 mg/L) and the TFA production (97.8 mg/L) was not very high and the DHA yield [2.63% chemical oxygen demand (COD)] and TFA yield (13.1% COD) were also very low. Furthermore, T. lutea using glycerol had a period of adaptation, indicating that T. lutea was not an ideal microalga for organic carbon utilization.
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We demonstrate that Mycobacterium tuberculosis recombinant leucine-responsive regulatory protein (rLrp) inhibits lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-α), interleukin-6, and interleukin-12 production and blocks the nuclear translocation of subunits of the nuclear-receptor transcription factor NF-κB (Nuclear factor-kappa B). Moreover, rLrp attenuated LPS-induced DNA binding and NF-κB transcriptional activity, which was accompanied by the degradation of inhibitory IκBα and a consequent decrease in the nuclear translocation of the NF-κB p65 subunit. RLrp interfered with the LPS-induced clustering of TNF receptor-associated factor 6 and with interleukin-1 receptor-associated kinase 1 binding to TAK1. Furthermore, rLrp did not attenuate proinflammatory cytokines or the expression of CD86 and major histocompatibility complex class-II induced by interferon-gamma in the macrophages of Toll-like receptor 2 deletion (TLR2-/-) mice and in protein kinase b (Akt)-depleted mouse cells, indicating that the inhibitory effects of rLrp were dependent on TLR2-mediated activation of the phosphatidylinositol 3-OH kinase (PI3K)/Akt pathway. RLrp could also activate the PI3K/Akt pathway by stimulating the rapid phosphorylation of PI3K, Akt, and glycogen synthase kinase 3 beta in macrophages. In addition, 19 amino acid residues in the N-terminus of rLrp were determined to be important and required for the inhibitory effects mediated by TLR2. The inhibitory function of these 19 amino acids of rLrp raises the possibility that mimetic inhibitory peptides could be used to restrict innate immune responses in situations in which prolonged TLR signaling has deleterious effects. Our study offers new insight into the inhibitory mechanisms by which the TLR2-mediated PI3K/Akt pathway ensures the transient expression of potent inflammatory mediators.
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Células Apresentadoras de Antígenos/imunologia , Proteínas de Bactérias/metabolismo , Citocinas/biossíntese , Regulação para Baixo , Mediadores da Inflamação/metabolismo , Macrófagos/metabolismo , Mycobacterium tuberculosis/metabolismo , Receptor 2 Toll-Like/metabolismo , Animais , Células Apresentadoras de Antígenos/efeitos dos fármacos , Autofagossomos/efeitos dos fármacos , Autofagossomos/metabolismo , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Regulação para Baixo/efeitos dos fármacos , Glicogênio Sintase Quinase 3 beta/metabolismo , Interferon gama/farmacologia , Quinases Associadas a Receptores de Interleucina-1 , Ligantes , Lipopolissacarídeos/farmacologia , MAP Quinase Quinase Quinases/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Ligação Proteica/efeitos dos fármacos , Subunidades Proteicas/metabolismo , Transporte Proteico/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacos , Fator 6 Associado a Receptor de TNF/metabolismoRESUMO
Brucella abortus is a zoonotic Gram-negative pathogen that causes brucelosis in ruminants and humans. Toll-like receptors (TLRs) recognize Brucella abortus and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. In this study, we focused on recombinant Brucella cell-surface protein 31 (rBCSP31) to determine its effects on mouse macrophages. Our results demonstrated that rBCSP31 induced TNF-α, IL-6 and IL-12p40 production, which depended on the activation of mitogen-activated protein kinases (MAPKs) by stimulating the rapid phosphorylation of p38 and JNK and the activation of transcription factor NF-κB in macrophages. In addition, continuous exposure (>24 h) of RAW264.7 cells to rBCSP31 significantly enhanced IFN-γ-induced expression of MHC-II and the ability to present rBCSP31 peptide to CD4(+) T cells. Furthermore, we found that rBCSP31 could interact with both TLR2 and TLR4. The rBCSP31-induced cytokine production by macrophages from TLR2(-/-) and TLR4(-/-) mice was lower than that from C57BL/6 macrophages, and the activation of NF-κB and MAPKs was attenuated in macrophages from TLR2(-/-) and TLR4(-/-) mice. In addition, CD4(+) T cells from C57BL/6 mice immunized with rBCSP31 produced higher levels of IFN-γ and IL-2 compared with CD4(+) T cells from TLR2(-/-) and TLR4(-/-) mice. Macrophages from immunized C57BL/6 mice produced higher levels of IL-12p40 than those from TLR2(-/-) and TLR4(-/-) mice. Furthermore, immunization with rBCSP31 provided better protection in C57BL/6 mice than in TLR2(-/-) and TLR4(-/-) mice after B. abortus 2308 challenge. These results indicate that rBCSP31 is a TLR2 and TLR4 agonist that induces cytokine production, upregulates macrophage function and induces the Th1 immune response.
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Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Brucella abortus/imunologia , Brucelose/imunologia , Macrófagos/imunologia , Células Th1/imunologia , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Apresentação de Antígeno/genética , Citocinas/metabolismo , Células HEK293 , Humanos , Mediadores da Inflamação/metabolismo , Ativação Linfocitária/genética , Sistema de Sinalização das MAP Quinases/genética , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Proteínas Recombinantes/imunologia , Células Th1/microbiologia , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genéticaRESUMO
We investigated whether a combined DNA vaccine delivered together with the IL-15 gene (DNA-IL-15(+)) enhanced the immune response against Brucella abortus in mice. Mice vaccinated with DNA-IL-15(+) developed a robust humoral response; Brucella-specific antibodies exhibited a dominance of immunoglobulin G2a (IgG2a) over IgG1. Splenocytes from DNA-IL-15(+)-vaccinated mice induced significantly higher levels of IFN-gamma (P<0.01) and CD8(+) T cell response (P<0.01), suggesting induction of a T-helper-1-dominated immune response. In a specific cytotoxic-T-lymphocyte activity assay, DNA-IL-15(+) immunization elicited mainly CD8(+) T cells, which mediate cytotoxicity, but also CD4(+) T cells. In vivo depletion of T cell subsets showed that the DNA-IL-15(+)-induced protection against Brucella infection is mediated predominantly by CD8(+) T cells, although CD4(+) T cells also contribute. These data indicate that plasmid-delivered IL-15 increases the efficacy of the Brucella DNA vaccine.
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Adjuvantes Imunológicos/administração & dosagem , Vacina contra Brucelose/imunologia , Linfócitos T CD8-Positivos/imunologia , Interleucina-15/administração & dosagem , Linfócitos T Citotóxicos/imunologia , Vacinas de DNA/imunologia , Animais , Anticorpos Antibacterianos/sangue , Vacina contra Brucelose/genética , Brucella abortus/genética , Brucella abortus/imunologia , Linfócitos T CD4-Positivos/imunologia , Testes Imunológicos de Citotoxicidade , Feminino , Imunoglobulina G/sangue , Interferon gama/metabolismo , Interleucina-15/genética , Interleucina-4/metabolismo , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos BALB C , Baço/imunologia , Baço/patologia , Subpopulações de Linfócitos T/imunologiaRESUMO
Bacterial community structure and the effects of several environmental factors on the microbial community distribution were investigated in the sediment of the eutrophic Lake Xuanwu. Profiles of bacterial communities were generated using denaturing gradient gel electrophoresis (DGGE), and the results were interpreted with multivariate statistical analysis. Five major variables in sediment were examined in a principal component analysis, which indicates notable differences of physicochemical parameters among different sites of the lake. To assess changes in the genetic diversity of bacterial communities of different sampling sites, DGGE band patterns were analyzed by multidimensional scaling analysis, which indicated that sampling sites having similar environmental characteristics also have the similar microbial communities. Canonical correspondence analysis demonstrated that pH and redox potential had significant effects on the bacterial community composition in the sediments. Analysis of DNA sequences revealed that the dominant bacterial groups in Lake Xuanwu belonged to Proteobacteria, Actinobacteria, Verrucomicrobia, and Nitrospirae, which are commonly isolated from freshwater ecosystems.