RESUMEN
Whole genome sequencing has revealed that the genome of Staphylococcus aureus possesses an uncharacterized 5-gene operon (SAOUHSC_00088-00092 in strain 8325 genome) that encodes factors with functions related to polysaccharide biosynthesis and export, indicating the existence of a new extracellular polysaccharide species. We designate this locus as ssc for staphylococcal surface carbohydrate. We found that the ssc genes were weakly expressed and highly repressed by the global regulator MgrA. To characterize Ssc, Ssc was heterologously expressed in Escherichia coli and extracted by heat treatment. Ssc was also conjugated to AcrA from Campylobacter jejuni in E. coli using protein glycan coupling technology (PGCT). Analysis of the heat-extracted Ssc and the purified Ssc-AcrA glycoconjugate by tandem mass spectrometry revealed that Ssc is likely a polymer consisting of N-acetylgalactosamine. We further demonstrated that the expression of the ssc genes in S. aureus affected phage adsorption and susceptibility, suggesting that Ssc is surface-exposed. IMPORTANCE: Surface polysaccharides play crucial roles in the biology and virulence of bacterial pathogens. Staphylococcus aureus produces four major types of polysaccharides that have been well-characterized. In this study, we identified a new surface polysaccharide containing N-acetylgalactosamine (GalNAc). This marks the first report of GalNAc-containing polysaccharide in S. aureus. Our discovery lays the groundwork for further investigations into the chemical structure, surface location, and role in pathogenesis of this new polysaccharide.
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Acetilgalactosamina , Polisacáridos Bacterianos , Staphylococcus aureus , Staphylococcus aureus/genética , Staphylococcus aureus/metabolismo , Acetilgalactosamina/metabolismo , Polisacáridos Bacterianos/metabolismo , Polisacáridos Bacterianos/genética , Polisacáridos Bacterianos/química , Regulación Bacteriana de la Expresión Génica , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Campylobacter jejuni/genética , Campylobacter jejuni/metabolismoRESUMEN
Globally, forest soils are considered as important sources and sinks of greenhouse gases (GHGs). However, most studies on forest soil GHG fluxes are confined to the topsoils (above 20 cm soil depths), with only very limited information being available regarding these fluxes in the subsoils (below 20 cm soil depths), especially in managed forests. This limits deeper understanding of the relative contributions of different soil depths to GHG fluxes and global warming potential (GWP). Here, we used a concentration gradient-based method to comprehensively investigate the effects of thinning intensity (15% vs. 35%) and nutrient addition (no fertilizer vs. NPK fertilizers) on soil GHG fluxes from the 0-40 cm soil layers at 10 cm depth intervals in a Chinese fir (Cunninghamia lanceolata) plantation. Results showed that forest soils were the sources of CO2 and N2O, but the sinks of CH4. Soil GHG fluxes decreased with increasing soil depth, with the 0-20 cm soil layers identified as the dominant producers of CO2 and N2O and consumers of CH4. Thinning intensity did not significantly affect soil GHG fluxes. However, fertilization significantly increased CO2 and N2O emissions and CH4 uptake at 0-20 cm soil layers, but decreased them at 20-40 cm soil layers. This is because fertilization alleviated microbial N limitation and decreased water filled pore space (WFPS) in topsoils, while it increased WFPS in subsoils, ultimately suggesting that soil WFPS and N availability (especially NH4+-N) were the predominant regulators of GHG fluxes along soil profiles. Generally, there were positive interactive effects of thinning and fertilization on soil GHG fluxes. Moreover, the 35% thinning intensity without fertilization had the lowest GWP among all treatments. Overall, our results suggest that fertilization may not only cause depth-dependent effects on GHG fluxes within soil profiles, but also impede efforts to mitigate climate change by promoting GHG emissions in managed forest plantations.
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Fertilizantes , Gases de Efecto Invernadero , Suelo , Gases de Efecto Invernadero/análisis , Suelo/química , Bosques , Metano/análisis , Dióxido de Carbono/análisis , Cunninghamia/crecimiento & desarrollo , Calentamiento Global , Óxido Nitroso/análisis , ChinaRESUMEN
Vermicompost is a promising amendment for immobilization of cadmium (Cd) in soils; however, its effectiveness can be influenced by rhizosphere environment conditions, such as pH and the presence of low-molecular-weight organic acids (LMWOAs). In this study, a batch experiment was conducted to examine the characteristics of Cd adsorption by vermicompost at different pH (pH = 3, 5, and 7) and after the addition of different LMWOAs (oxalic acid; citric acid; malic acid). Furthermore, a series of morphology and structural analyses were conducted to elucidate the mechanisms of observed effects. The results showed that the adsorption capacity of vermicompost for Cd increased as pH increased, and chemisorption dominated the adsorption process. Changes in pH altered adsorption performance by affecting the -OH groups of alcohol/phenol and the -CH2 groups of aliphatics. Further, the addition of oxalic acid promoted Cd adsorption, and the effect was concentration dependent. Modifying the verimicompost surface with more adsorption sites might be the main reason. Conversely, citric acid and malic acid showed the ability to inhibit Cd adsorption by vermicompost. Citric acid caused a blocking effect by covering flocculent substances on the vermicompost surface while reducing surface adsorption sites by dissolving mineral components such as iron oxides. However, the action of malic acid did not appear to be related to changes in morphology or the structure of vermicompost. Overall, the results of this study partially explain the limited effectiveness of Cd immobilization within the rhizosphere by vermicompost, and provide theoretical support for regulating rhizosphere environments to improve the effectiveness of vermicompost immobilization of Cd.
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Cadmio , Contaminantes del Suelo , Cadmio/análisis , Adsorción , Rizosfera , Suelo/química , Compuestos Orgánicos , Ácido Oxálico/química , Ácido Cítrico/química , Concentración de Iones de Hidrógeno , Contaminantes del Suelo/análisisRESUMEN
Evaluating heavy metal pollution level in the soils and apportioning the source-specific health risk of heavy metals are critical for proposing environmental protection and remediation strategies to protection human health. This study explored heavy metal pollution and associated source-specific health risks in a typical rural industrial area, southwestern China. A total of 105 topsoil samples were collected and the concentrations of heavy metals, including As, Cd, Cr, Cu, Ni, Pb and Zn, were determined. Pollution load index and enrichment factors were used to evaluate the pollution level of heavy metals. Positive matrix factorization (PMF) model was applied to apportion the heavy metals and the associated source-specific health risks to adults and children were estimated via combining the PMF model with the health risk assessment. The results indicated that the soils were highly polluted by multiple heavy metals, especially for Cd, with the EF values of 24.94 and 22.55 in the upstream and downstream areas, respectively. Source apportionment results showed that atmospheric deposition, smelting activities, fertilizer and sewage application, and agrochemical utilization were the main anthropogenic sources, with the contributions of 37.11%, 23.69%, 19.69%, and 19.51%, respectively. Source-specific risk assessment identified atmospheric deposition as the priority source for the non-carcinogenic (NCR) and carcinogenic risks (CR) in the study area, with the contribution of 43.71% and 52.52% for adults, and 44.29% and 52.58% for children, respectively. Moreover, non-carcinogenic and carcinogenic risks posed to children (NCR: 2.84; CR: 1.31 × 10-4) from four sources was higher than those posed to adults (NCR: 0.29; CR: 5.86 × 10-5). The results of source-specific health risk assessment provided the valuable information on the priority sources for pollution preventing and risk controlling.
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Metales Pesados , Contaminantes del Suelo , Adulto , Niño , Humanos , Suelo , Monitoreo del Ambiente , Cadmio/análisis , Incertidumbre , Contaminantes del Suelo/análisis , Metales Pesados/toxicidad , Metales Pesados/análisisRESUMEN
Production of capsular polysaccharides in Staphylococcus aureus is transcriptionally regulated by a control region of the cap operon that consists of SigA- and SigB-dependent promoters. A large number of regulators have been shown to affect cap gene expression. However, regulation of capsule is only partially understood. Here we found that SarZ was another regulator that activated the cap genes through the SigA-dependent promoter. Gel electrophoresis mobility shift experiments revealed that SarZ is bound to a broad region of the cap promoter including the SigA-dependent promoter but mainly the downstream region. We demonstrated that activation of cap expression by SarZ was independent of MgrA, which also activated capsule through the SigA-dependent promoter. Our results further showed that oxidative stress with hydrogen peroxide (H2O2) treatments enhanced SarZ activation of cap expression, indicating that SarZ is able to sense oxidative stress to regulate capsule production. IMPORTANCE Expression of virulence genes in Staphylococcus aureus is affected by environmental cues and is regulated by a surprisingly large number of regulators. Much is still unknown about how virulence factors are regulated by environment cues at the molecular level. Capsule is an antiphagocytic virulence factor that is highly regulated. In this study, we found SarZ was an activator of capsule and that the regulation of capsule by SarZ was affected by oxidative stress. These results provide an example of how a virulence factor could be regulated in response to an environmental cue. As the host oxidative defense system plays an important role against S. aureus, this study contributes to a better understanding of virulence gene regulation and staphylococcal pathogenesis.
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Infecciones Estafilocócicas , Staphylococcus aureus , Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Humanos , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , Inmunoglobulina A Secretora/genética , Staphylococcus aureus/metabolismo , Factores de Virulencia/metabolismoRESUMEN
Petrochemical industry is a key industry of soil pollution, which presents great effects on human health and the ecological environment. It is of great significance to achieve rapid, economic and efficient health risk identification for petrochemical industry in China. In this work, an efficient method was developed based on extreme gradient boosting (XGBoost) algorithm for human health risk identification, which is different from the traditional health risk assessment with complicated procedures. In this methodology, an index system of 13 indicators was established from the perspective of "sources - pathways - receptors" for risk identification. The 10-fold cross validation was used to assess the generalization performance, and the accuracy, precision and recall were employed to evaluate the performance of the algorithms. Wilcoxon signed-rank test was conducted to analyze the differences between XGBoost and other models for statistical support. The results showed that XGBoost significantly presented a better performance for health risk identification over multilayer perceptron neural network with error backpropagation training (BPNN), support vector machine (SVM), gradient boosting decision tree (GBDT) and light gradient boosting machine (LightGBM), with an accuracy of 0.783. The most important features contributing to the risk identification were determined with the sequence of site location (in the industrial zone or not), site planning and production period. Great attention should be given to the petrochemical sites that are not located in the industrial zone with long production period and sensitive receptors in the health risk identification. This method has important reference significance for relevant departments to carry out soil contamination screening and health risk assessment of petrochemical sites.
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Redes Neurales de la Computación , Máquina de Vectores de Soporte , Algoritmos , Humanos , Medición de Riesgo , SueloRESUMEN
Soil pollution at industrial sites is an important issue in China and in most other regions of the world. The accurate prediction of the spatial distribution of pollutants at contaminated industrial sites is a requirement for the development of most soil remediation strategies, and is commonly performed using spatial interpolation methods. However, significant and abrupt variations in the spatial distribution of pollutants decrease prediction accuracy. During this study, the use of partition interpolation methods was applied to benzo fluoranthene in four soil layers at a contaminated site to determine their ability to improve prediction accuracy in comparison to unpartitioned methods. The examined methods for partitioned interpolation included inverse distance weighting (IDW), radial basis function (RBF), and ordinary kriging (OK). The prediction results of the three methods for partitioned interpolation were compared, and the applicability of partition interpolation was determined. The prediction error associated with the partitioned interpolation methods decreased by 70% compared to unpartitioned interpolation. The prediction accuracy of IDW-based partition interpolation was higher than that of RBF- and OK-based partition interpolation techniques, and it was suitable for identification of highly polluted areas. Partition interpolation is also applicable to 12 other PAHs controlled by USEPA that can be detected, and the prediction effects could also verify this interpolation choice. In addition, the results also demonstrated that the more the maximum concentration deviated from the "norm", the greater the prediction error was caused by the smoothing effects of the interpolation models. These results suggest that the partition interpolation with IDW method can be effectively used to obtain relatively accurate spatial contaminant distribution information, and to identify highly polluted areas.
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Monitoreo del Ambiente/métodos , Contaminantes del Suelo/análisis , China , Contaminación Ambiental/análisis , Contaminación Ambiental/prevención & control , Fluorenos/análisis , Predicción , Análisis EspacialRESUMEN
To study the effect and mechanism of extract of Quzhou Aurantii Fructus(QAF) on liver inflammation in CCl_4-induced liver fibrosis mice. Totally 60 C57 BL/6 male mice were randomly divided into control group(distilled water, oral), model group(distilled water, oral), colchicines group(Col, colchicines 2 mg·kg~(-1)·d~(-1), oral), low-dose QAF group(QAF-L, QAF 100 mg·kg~(-1)·d~(-1), oral) and high-dose QAF group(QAF-H, QAF 300 mg·kg~(-1)·d~(-1), oral) by random number table method. The model group and each administration group were injected with carbon tetrachloride(CCl_4) 1 mL·kg~(-1)(CCl_4-olive oil 1â¶4), twice a week, totally 6 weeks. After the last administration, the mice were sacrificed, and serum and liver tissue were collected. Serum ALT and AST levels were measured in each group to observe the liver function of mice. The pathological changes and inflammatory cell infiltration in liver were observed by HE staining and F4/80 immunohistochemical staining. The mRNA expressions of TNF-α, IL-18 and IL-1ß were detected by RT-PCR. The protein expressions of IκBα, p-IKKα/ß, p-p65, NLRP3, caspase-1 and cleaved caspase-1 were analyzed by Western blot. The results showed that QAF significantly reduced serum ALT and AST levels, and alleviated the degree of liver damage.The results of immunohistochemistry showed that QAF significantly reduced liver inflammatory cell infiltration in liver fibrosis mice. The results of RT-PCR and Western blot showed that QAF significantly inhibited mRNA expressions of TNF-α, IL-18 and IL-1ß in liver of fibrosis mice. QAF also suppressed the degradation of IκBα protein and reduced p-IKKα/ß, p-p65, NLRP3 and cleaved caspase-1 protein expressions. In conclusion, QAF improves CCl_4-induced liver fibrosis in mice. The mechanism may be related to the inhibition of NF-κB/NLRP3 inflammasome-mediated inflammation signaling pathway.
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Inflamasomas , FN-kappa B , Animales , Inflamasomas/genética , Inflamación , Hígado/patología , Cirrosis Hepática/inducido químicamente , Cirrosis Hepática/tratamiento farmacológico , Cirrosis Hepática/genética , Masculino , Ratones , FN-kappa B/genética , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Extractos VegetalesRESUMEN
Medicinal plants are beneficial to human health. However,most of the major producing regions of medicinal plants suffer from rust disease,which threatens the yield and quality of Chinese medicinal materials,thus causes huge economic loss,and hinders the sustainable development of the Chinese medicine industry. By the end of 2020,rust disease had been reported in medicinal plants of 76 species and 33 families. In the 76 species,79 rust pathogens were detected. The majority of these pathogens belonged to Puccinia( 33,39. 24%),Coleosporium( 14,15. 19%),and Aecidium( 11,13. 92%). Of these 79 rust pathogens,10 were autoecious and 13 were heteroecious. Through literature research,this study reviewed the symptoms,pathogen species,severity and distribution,prevalence and occurrence conditions,and control measures of rust disease in medicinal plants,and thereby summarized the research status of rust disease in medicinal plants and the gap with other plants,which is expected to serve as a reference for further research on rust disease in medicinal plants.
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Basidiomycota , Plantas Medicinales , Basidiomycota/genética , Humanos , Enfermedades de las PlantasRESUMEN
Staphylococcus aureus capsule polysaccharide is an important antiphagocytic virulence factor. The cap genes are regulated at the promoter element (Pcap) upstream of the cap operon. Pcap, which consists of a dominant SigB-dependent promoter and a weaker upstream SigA-dependent promoter, is activated by global regulator MgrA. How MgrA activates capsule is unclear. Here, we showed that MgrA directly bound to the Pcap region and affected the SigA-dependent promoter. Interestingly, an electrophoretic mobility shift assay showed that MgrA bound to a large region of Pcap, mainly downstream of the SigA-dependent promoter. We further showed that the ArlRS two-component system and the Agr quorum sensing system activated capsule primarily through MgrA in the early growth phases.IMPORTANCE The virulence of Staphylococcus aureus depends on the expression of various virulence factors, which is governed by a complex regulatory network. We have been using capsule as a model virulence factor to study virulence gene regulation in S. aureus MgrA is one of the regulators of capsule and has a major effect on capsule production. However, how MgrA regulates capsule genes is not understood. In this study, we were able to define the mechanism involving MgrA regulation of capsule. In addition, we also delineated the role of MgrA in capsule regulatory pathways involving the key virulence regulators Agr and Arl. This study further advances our understanding of virulence gene regulation in S. aureus, an important human pathogen.
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Cápsulas Bacterianas/química , Inmunoglobulina A Secretora/fisiología , Polisacáridos Bacterianos/fisiología , Regiones Promotoras Genéticas/fisiología , Staphylococcus aureus/fisiología , Factores de Virulencia/fisiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Ensayo de Cambio de Movilidad Electroforética , Immunoblotting , Inmunoglobulina A Secretora/genética , Mutación , Polisacáridos Bacterianos/genética , ARN Bacteriano/aislamiento & purificación , ARN Bacteriano/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Transcripción Reversa , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidad , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
Soft tissue leiomyosarcoma (STLMS) is a major histological subtype of adult sarcoma. Although the molecular mechanisms ofLMS have been gradually revealed, no valid therapeutic targets have been identified. In this study, we performed a systematic screening to explore relapse-associated genes in STLMS, using data from The Cancer Genome Atlas-Sarcoma (TCGA-SARC). Then, we investigated the functional role of the gene with the best relapse-prediction value in STLMS by both in-vitro and in-vivo studies. Results showed that AMH and PLA2G10 were two genes with area under curve (AUC) values higher than 0.80 in ROC analysis when detecting relapse. Patients in the high AMH or PLA2G10 expression group had significantly worse relapse-free survival (RFS) compared to the respective low expression group. PLA2G10 was highly expressed in STLMS, but not in other sarcoma subtypes. PLA2G10 overexpression promoted SK-LMS-1 cell growth and G1/S transition, while PLA2G10 knockdown slowed the growth and resulted in G1 phase arrest. PLA2G10 overexpression markedly increased the expression of CDK2 and cyclin E1, but did not influence CDK4, CDK6, cyclin D1, CDK1 or cyclin A expression. PLA2G10 overexpression enhanced SK-LMS-1 cell-derived xenograft tumor growth in nude mice, while PLA2G10 inhibition slowed the growth. Mutation of two critical catalyzing amino acid residues (p.H88A and p.D89A) abrogated the capability of PLA2G10 to catalyze the production of arachidonic acid (AA), and also canceled the regulatory effects on cyclin E1 and CDK2 expression, as well as G1/S transition. In conclusion, PLA2G10 was a specific relapse-associated gene in STLMS. It facilitated the cell-cycle progression of STLMS cells at least by elevating the expression of cyclin E1 and CDK2. The hydrolytic activity was crucial for its oncogenic properties.
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Ciclina E/genética , Quinasa 2 Dependiente de la Ciclina/genética , Regulación Neoplásica de la Expresión Génica , Fosfolipasas A2 Grupo X/genética , Leiomiosarcoma/genética , Proteínas Oncogénicas/genética , Animales , Ciclo Celular , Línea Celular Tumoral , Femenino , Humanos , Leiomiosarcoma/patología , Ratones Endogámicos BALB C , Ratones Desnudos , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/patologíaRESUMEN
Clinical data have shown that pulmonary interstitial fibrosis is likely to occur in the later stages of viral pneumonia. While viral infections are thought to cause chronic pulmonary interstitial inflammation and pulmonary fibrosis, it remains unclear if they promote pulmonary fibrosis by epithelial-mesenchymal transition (EMT). In this study, human epithelial cell line A549 has been used to model the infection of the Epstein-Barr virus (EBV) and the respiratory syncytial virus (RSV). Their differences were compared and the possible infection mechanisms analyzed by randomly assigning cells to one of five treatments. Exposure of the LMP1 is thought to be the key gene during EBV-induced EMT in the A549 cells. Enzyme-linked immunosorbent assay analysis revealed that the EBV infection was associated with the induction of a number of cytokines (interleukin-8 [IL-8], IL-13, tumor necrosis factor-α, and transforming growth factor-ß) and dexamethasone (DXM) could significantly prevent the phenotypic changes, and partly the mechanisms related with the IL-13 pathway. Surprisingly, different results were seen with the RSV infection as the A549 cells still displayed an epithelial morphology but the levels of E-cadherin, α-SMA, vimentin, and fibronectin did not change. This is the first study demonstrating the different reactions induced by different viruses, and the protective effects of DXM on the EBV-induced EMT in the A549 cells by partially inhibiting the IL-13 pathway. These findings suggest a novel mechanism, by which DXM or anti-IL-13 may delay the progression of pulmonary fibrosis by preventing the progress of EBV-induced EMT.
RESUMEN
The fern Pteris vittata has been the subject of numerous studies because of its extreme arsenic hyperaccumulation characteristics. However, information on the arsenic chemical speciation and distribution across cell types within intact frozen-hydrated Pteris vittata fronds is necessary to better understand the arsenic biotransformation pathways in this unusual fern. While 2D X-ray absorption spectroscopy imaging studies show that different chemical forms of arsenic, As(III) and As(V), occur across the plant organs, depth-resolved information on arsenic distribution and chemical speciation in different cell types within tissues of Pteris vittata have not been reported. By using a combination of planar and confocal µ-X-ray fluorescence imaging and fluorescence computed µ-tomography, we reveal, in this study, the localization of arsenic in the endodermis and pericycle surrounding the vascular bundles in the rachis and the pinnules of the fern. Arsenic is also accumulated in the vascular bundles connecting into each sporangium, and in some mature sori. The use of 2D X-ray absorption near edge structure imaging allows for deciphering arsenic speciation across the tissues, revealing arsenate in the vascular bundles and arsenite in the endodermis and pericycle. This study demonstrates how different advanced synchrotron X-ray microscopy techniques can be complementary in revealing, at tissue and cellular levels, elemental distribution and chemical speciation in hyperaccumulator plants.
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Arsénico , Pteris , Contaminantes del Suelo , Tomografía Computarizada por Rayos X , Espectroscopía de Absorción de Rayos XRESUMEN
Depression is a progressive and chronic syndrome and commonly related to several neuropsychiatric comorbidities, of which depression is the most studied. Population-based studies have suggested a positive role of statins in ameliorating depression risk. However, the role of statins in the treatment of diabetes-related depression has not been well examined. Herein, we investigated the effects of lovastatin (LOV) on depressive phenotypes in streptozotocin-induced diabetic mice. The data suggested that the treatment of LOV at 10 or 20 mg/kg for 3 weeks markedly prevented diabetes-associated depressive behaviors reflected by better performance in the sucrose preference test, tail suspension test, and novelty-suppressed feeding test. The study further showed that these treatments improved the hippocampal neurogenesis as evidenced by increased bromodeoxyuridine-positive cells in the dentate gyrus with higher expression of mature brain-derived neurotrophic factor and increased phosphorylation of cAMP-response element-binding protein. As expected, diabetic mice treated with LOV showed significant improvement of hyperlipidemia rather than hyperglycemia. These results suggest that LOV may be employed as a drug for the treatment of diabetes-related depression.
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Depresión/prevención & control , Diabetes Mellitus Experimental/complicaciones , Hipocampo/efectos de los fármacos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Lovastatina/uso terapéutico , Neurogénesis/efectos de los fármacos , Animales , Antidepresivos , Conducta Animal/efectos de los fármacos , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Depresión/etiología , Diabetes Mellitus Experimental/inducido químicamente , Masculino , Ratones , Ratones Endogámicos C57BL , Estrés PsicológicoRESUMEN
Information on the kinetic characteristics of soil enzymes under long-term arsenic (As) pollution in field soils is scarce. We investigated Michaelis-Menten kinetic properties of four soil enzymes including ß-glucosidase (BG), acid phosphatase (ACP), alkaline phosphatase (ALP), and dehydrogenase (DHA) in field soils contaminated by As resulting from long-term realgar mining activity. The kinetic parameters, namely the maximum reaction velocity (Vmax), enzyme-substrate affinity (Km) and catalytic efficiency (Vmax/Km) were calculated. Results revealed that the enzyme kinetic characteristics varied in soils and were significantly influenced by total nitrogen (N) and total As, which explained 31.8% and 30.7% of the variance in enzyme kinetics respectively. Enzyme pools (Vmax) and catalytic efficiency (Vmax/Km) of BG, ACP and DHA decreased with elevated As pollution, while the enzyme affinity for substrate (Km) was less affected. Redundancy analysis and stepwise regression suggested that the adverse influence of As on enzyme kinetics may offset or weakened by soil total N and soil organic matter (SOM). Concentration-response fitting revealed that the specific kinetic parameters expressed as the absolute enzyme kinetic parameters multiplied by normalized soil total N and SOM were more relevant than the absolute ones to soil total As. The arsenic ecological dose values that cause 10% decrease (ED10) in the specific enzyme kinetics were 20-49 mg kg-1, with a mean value of 35 mg kg-1, indicating a practical range of threshold for As contamination at field level. This study concluded that soil enzymes exhibited functional adaptation to long-term As stress mainly through the reduction of enzyme pools (Vmax) or maintenance of enzyme-substrate affinity (Km). Further, this study demonstrates that the specific enzyme kinetics are the better indicators of As ecotoxicity at field-scale compared with the absolute enzyme parameters.
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Arsénico/toxicidad , Enzimas/química , Contaminantes del Suelo/toxicidad , Suelo/química , Arsénico/análisis , Arsenicales , Biomarcadores Ambientales/efectos de los fármacos , Cinética , Minería , Nitrógeno/análisis , Compuestos Orgánicos/análisis , Contaminantes del Suelo/análisis , Sulfuros/toxicidadRESUMEN
The SaeRS two-component system in Staphylococcus aureus is critical for regulation of many virulence genes, including hla, which encodes alpha-toxin. However, the impact of regulation of alpha-toxin by Sae on S. aureus pathogenesis has not been directly addressed. Here, we mutated the SaeR-binding sequences in the hla regulatory region and determined the contribution of this mutation to hla expression and pathogenesis in strain USA300 JE2. Western blot analyses revealed drastic reduction of alpha-toxin levels in the culture supernatants of SaeR-binding mutant in contrast to the marked alpha-toxin production in the wild type. The SaeR-binding mutation had no significant effect on alpha-toxin regulation by Agr, MgrA, and CcpA. In animal studies, we found that the SaeR-binding mutation did not contribute to USA300 JE2 pathogenesis using a rat infective endocarditis model. However, in a rat skin and soft tissue infection model, the abscesses on rats infected with the mutant were significantly smaller than the abscesses on those infected with the wild type but similar to the abscesses on those infected with a saeR mutant. These studies indicated that there is a direct effect of hla regulation by SaeR on pathogenesis but that the effect depends on the animal model used.
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Proteínas Bacterianas/fisiología , Toxinas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Proteínas Quinasas/fisiología , Infecciones Estafilocócicas , Staphylococcus aureus , Animales , Toxinas Bacterianas/genética , Ratas , Infecciones Estafilocócicas/genética , Infecciones Estafilocócicas/metabolismo , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidad , VirulenciaRESUMEN
Virulence genes are regulated by a complex regulatory network in Staphylococcus aureus Some of the regulators are global in nature and affect many downstream genes. MgrA is a multiple-gene regulator that has been shown to activate genes involved in capsule biosynthesis and repress surface protein genes. The goal of this study was to demonstrate the biological significance of MgrA regulation of capsule and surface proteins. We found that strain Becker possessed one fibronectin-binding protein, FnbA, and that FnbA was the predominant protein involved in invasion of nonphagocytic HeLa cells. By genetic analysis of strains with different amounts of capsule, we demonstrated that capsule impeded invasion of HeLa cells by masking the bacterial cell wall-anchored protein FnbA. Using variants with different levels of mgrA transcription, we further demonstrated that MgrA negatively impacted invasion by activating the cap genes involved in capsule biosynthesis and repressing the fnbA gene. Thus, we conclude that MgrA negatively impacts cell invasion of S. aureus Becker by promoting capsule and repressing FnbA.
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Adhesinas Bacterianas/metabolismo , Cápsulas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica/genética , Infecciones Estafilocócicas/patología , Staphylococcus aureus/patogenicidad , Adhesinas Bacterianas/genética , Cápsulas Bacterianas/genética , Línea Celular Tumoral , Células HeLa , Humanos , Polisacáridos Bacterianos/metabolismo , Infecciones Estafilocócicas/genética , Staphylococcus aureus/genética , Virulencia/genéticaRESUMEN
Soil water is a major interference in the on-site analysis of soil by energy-dispersive X-ray fluorescence. Apparent consequences of this interference include lowered readings for elemental concentrations and significant changes in spectral characteristics in wet soils compared with dry soils. A rigorous interpretation on this issue remains unresolved. Thus, this study evaluated the impact of soil water on the detection of Ca, Ti, Mn, Fe, Cu, Zn, As, Rb, Sr, and Pb. Specimens were prepared from 11 certified reference soils and 3 field soils with water contents from 0 to â¼40 wt %. Results from three commercial models revealed that the readings were subjected to respective internal quantification algorithms; therefore, they could not provide a fundamental perspective of this issue. We analyzed the spectra to examine the mechanism underlying this phenomenon. The spectra of wet soils feature elevated baseline, increased Compton and Rayleigh scatter peaks, and lowered characteristic peaks of elements. Previous studies attributed the lowered characteristic peaks to the absorption of fluorescent X-rays by water and considered soil water and dry soil as separate layers in the calculation. This work argues that wet soils should be treated as mixtures. Water becomes part of the soil matrix and leads to lower attenuation capability, which could be explained by the matrix effect. Meanwhile, the mass fraction of analytes is lowered because of dilution. Results confirmed that dilution lowers the characteristic peaks, whereas the matrix effect heightens them. When estimating the elemental concentrations on a wet weight basis, the matrix effect becomes the major interference. The Compton compensation method provided satisfying results on correcting the matrix effect caused by soil water on Zn, As, Rb, Sr, and Pb.
Asunto(s)
Anomalías Cardiovasculares , Arteria Subclavia , Humanos , Arteria Subclavia/diagnóstico por imagen , Anomalías Cardiovasculares/complicaciones , Anomalías Cardiovasculares/diagnóstico por imagen , Anomalías Cardiovasculares/cirugía , Aorta Torácica , Arteria Carótida Común/diagnóstico por imagenRESUMEN
Accurate prediction of the spatial distribution of pollutants in soils based on applicable interpolation methods is often the basis for soil remediation in contaminated sites. However, the applicable interpolation method has not been determined for contaminated sites due to the complex spatial distribution characteristics and stronger local spatial variability of pollutants. In this research, the prediction accuracies of three interpolation methods (including the different values of their parameters) for the spatial distribution of benzo[b]fluoranthene (BbF) in four soil layers were compared. These included inverse distance weighting (IDW), radial basis function (RBF), ordinary kriging (OK). The results indicated: (1) IDW1 is applicable for the first layer, RBF-IMQ is applicable to the second, third, and fourth layers. (2) For IDW, the prediction error is bigger with high weight where high values and low values intersect, while the prediction error is smaller where high (or low) values aggregated distribution. (3) For RBF, if the pollutant concentration trend at the predicted location is consistent with the known points in its neighborhood, the prediction accuracy is higher. (4) IDW is suitable for fitting more drastic curved surfaces, while RBF is more effective for relatively gentle curved surfaces and OK is reasonable for curved surfaces without local outliers. (5) The interpolation uncertainty is positively associated with the contaminant concentration and local spatial variability. Therefore, we suggest the selection of the applicable interpolation model must be based on the principle of the model and the spatial distribution characteristics of the pollutants.