RESUMO
Polycyclic aromatic hydrocarbons (PAHs) and dioxins are potential causes of multiple diseases by activating the aryl hydrocarbon receptor (AhR) pathway. Health risk assessment of chemicals primarily relies on the relative potency factor (RPF), although its accuracy may be limited when solely using EC50 values. The induction of cytochrome P4501A1 (CYP1A1) serves as a biomarker for AhR activation and is an integrator of dioxin-like toxicity. Here, we present a method for evaluating the risks associated with AhR activation using mathematical models of dose-CYP1A1 induction. The dose-effect curves for certain PAHs and dioxins, including Ant, BghiP, 1,2,3,4,7,8-HxCDD, and others, exhibited a non-classical S-shaped form. The toxic equivalent factor (TEF) profiles revealed a broad range of toxic equivalent factor values. The TEFs for PAHs ranged from approximately 0.01 to 6, with higher values being observed when the concentration was less than 10-10 M, with the exceptions of Ace, Phe, and BghiP. Most congeners of dioxins got the lowest TEF value at around 10-10 M, ranging from 0.04 to 1.00. The binding affinity of AhR to ligands did not display a strong correlation with the EC50 of CYP1A1 expression, suggesting that the AhR-mediated effects of PAHs and dioxins are not fixed but instead fluctuate with the dose. Air samples acquired from a parking area were used to compare the proficiency of RPF and our current approach. In the current method, naphthalene and chrysene were the primary contributors of PAHs to AhR-mediated risks in parking lots air samples, respectively. However, the contributions of naphthalene and chrysene could be disregarded in the RPF approach.
Assuntos
Biomarcadores , Citocromo P-450 CYP1A1 , Dioxinas , Exposição por Inalação , Hidrocarbonetos Policíclicos Aromáticos , Receptores de Hidrocarboneto Arílico , Receptores de Hidrocarboneto Arílico/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Biomarcadores/metabolismo , Biomarcadores/análise , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/análise , Dioxinas/toxicidade , Medição de Risco , Humanos , Relação Dose-Resposta a DrogaRESUMO
BACKGROUND: Pregnancy-induced Cushing's syndrome (CS) with an adrenocortical adenoma overexpressing luteinizing hormone (LH)/human choriogonadotropin (hCG) receptors (LHCGR) has been rarely reported in the literatures. This peculiar condition challenges the canonical diagnosis and management of CS. CASE PRESENTATION: A 27-year-old woman (G2P0A1) presented at 20 weeks gestational age (GA) with overt Cushingoid clinical features. Adrenocorticotropic hormone (ACTH)-independent CS was diagnosed based on undetectable ACTH and unsuppressed cortisol levels by dexamethasone. Magnetic resonance imaging (MRI) scanning without contrast revealed a left adrenal nodule while pituitary MRI scanning was normal. A conservative treatment strategy of controlling Cushingoid comorbidities was conducted. At 36 weeks GA, a caesarean operation was performed and a live female infant was delivered. At 8 weeks after parturition, our patient achieved normalization of blood pressure, blood glucose, serum potassium, and urinary cortisol level spontaneously. During non-pregnancy period, stimulation testing with exogenous hCG significantly evoked a cortisol increase. The woman underwent resection of the adrenal tumor at 6 months after parturition. Immunohistochemistry (IHC) showed the tumor tissue that stained positive for luteinizing hormone (LH)/human choriogonadotropin (hCG) receptor (LHCGR), whereas negative for both melanocortin 2 receptor (MC2R) and G protein-coupled receptor-1 (GPER-1). CONCLUSIONS: Stimulation test with exogenous hCG after parturition is necessary for the diagnosis of pregnancy-induced CS. LHCGR plays an essential role in the pathogenesis of this rare condition.
Assuntos
Neoplasias do Córtex Suprarrenal/diagnóstico por imagem , Adenoma Adrenocortical/diagnóstico por imagem , Síndrome de Cushing/diagnóstico , Complicações Neoplásicas na Gravidez/diagnóstico por imagem , Receptores do LH/metabolismo , Neoplasias do Córtex Suprarrenal/complicações , Neoplasias do Córtex Suprarrenal/metabolismo , Neoplasias do Córtex Suprarrenal/cirurgia , Adenoma Adrenocortical/complicações , Adenoma Adrenocortical/metabolismo , Adenoma Adrenocortical/cirurgia , Adulto , Síndrome de Cushing/etiologia , Síndrome de Cushing/metabolismo , Técnicas de Diagnóstico Endócrino , Feminino , Humanos , Imageamento por Ressonância Magnética , Hipófise/diagnóstico por imagem , Gravidez , Complicações na Gravidez/diagnóstico , Complicações na Gravidez/etiologia , Complicações na Gravidez/metabolismo , Complicações Neoplásicas na Gravidez/metabolismo , Complicações Neoplásicas na Gravidez/cirurgiaRESUMO
Operation performance and bacterial community structure of sulfur-based autotrophic denitrification (SAD) based on different sulfur sources served as electron donor was first parallelly compared among three sequencing batch reactors. Sulfur and sodium thiosulfate systems achieved similar operation performance and were superior to that of sodium sulfide. When the influent NO3--N concentration ranged from 50 to 150 mg/L, the effluent NO3--N concentrations of the sulfur and sodium thiosulfate systems were 0-5.99 mg/L and 0-4.52 mg/L, respectively, without NO2--N accumulation. However, when the effluent concentration of NO3--N in the sodium sulfide system was 0-10.38 mg/L, that of NO2--N in the effluent was 0-39.85 mg/L. In addition, participation of sulfur sources presented obvious pressure on the bacterial community structure based on the high-throughput sequencing. Microbial diversity results indicated that sludge with elemental sulfur as electron donor had the richest microbial diversity, followed by sodium thiosulfate and sodium sulfide. Moreover, sludge with elemental sulfur and sodium thiosulfate as electron donor demonstrated more similar community structure compared with the sludge that denitrified with sodium sulfide according to the microbial similarity analysis. The 9.34%, 24.3% and 29.6% of sequences could be assigned to potential SAD organisms from sludge denitrifying with elemental sulfur, sodium thiosulfate and sodium sulfide, respectively. Furthermore, all sludge denitrifying with different sulfur sources showed an enrichment of separate core functional microorganisms. This study could provide an insight into improving the understanding of SAD in engineering applications.
Assuntos
Microbiota , Esgotos/microbiologia , Enxofre/metabolismo , Processos Autotróficos , Reatores Biológicos , Desnitrificação , Microbiota/genética , Nitratos/metabolismo , Esgotos/química , Sulfetos/metabolismo , Tiossulfatos/metabolismoRESUMO
Current studies on microbial isoprene biosynthesis have mostly focused on regulation of the upstream mevalonic acid (MVA) or methyl-erythritol-4-phosphate (MEP) pathway. However, the downstream bottleneck restricting isoprene biosynthesis capacity caused by the weak expression and low activity of plant isoprene synthase (ISPS) under microbial fermentation conditions remains to be alleviated. Here, based on a previously constructed Saccharomyces cerevisiae strain with enhanced precursor supply, we strengthened the downstream pathway through increasing both the expression and activity of ISPS to further improve isoprene production. Firstly, a two-level expression enhancement system was developed for the PGAL1-controlled ISPS by overexpression of GAL 4. Meanwhile, the native GAL1/7/10 promoters were deleted to avoid competition for the transcriptional activator Gal4p, and GAL80 was disrupted to eliminate the dependency of gene expression on galactose induction. The IspS expression was obviously elevated upon enhanced Gal4p supply, and the isoprene production was improved from 6.0mg/L to 23.6mg/L in sealed-vial cultures with sucrose as carbon source. Subsequently, a novel high-throughput screening method was developed based on precursor toxicity and used for ISPS directed evolution towards enhanced catalytic activity. Combinatorial mutagenesis of the resulting ISPS mutants generated the best mutant ISPSM4, introduction of which into the GAL4-overexpressing strain YXM29 achieved 50.2mg/L of isoprene in sealed vials, and the isoprene production reached 640mg/L and 3.7g/L in aerobic batch and fed-batch fermentations, respectively. These results demonstrated the effectiveness of the proposed combinatorial engineering strategy in isoprene biosynthesis, which might also be feasible and instructive for biotechnological production of other valuable chemicals.
Assuntos
Alquil e Aril Transferases/metabolismo , Butadienos/metabolismo , Proteínas de Ligação a DNA/metabolismo , Evolução Molecular Direcionada/métodos , Hemiterpenos/metabolismo , Engenharia Metabólica/métodos , Pentanos/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/fisiologia , Fatores de Transcrição/metabolismo , Vias Biossintéticas/genética , Butadienos/isolamento & purificação , Proteínas de Ligação a DNA/genética , Regulação Bacteriana da Expressão Gênica/genética , Regulação Enzimológica da Expressão Gênica/genética , Melhoramento Genético/métodos , Hemiterpenos/isolamento & purificação , Redes e Vias Metabólicas/genética , Pentanos/isolamento & purificação , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Fatores de Transcrição/genética , Regulação para Cima/genéticaRESUMO
Isoprene is facing a growing global market due to its wide industrial applications. Current industrial production of isoprene is almost entirely petroleum-based, which is influenced by the shrinking C5 supply, while the natural emission of isoprene is predominantly contributed by plants. To bridge the need gap, a highly efficient fermentation-based process for isoprene production might be a suitable and sustainable solution, and extensive research works have been performed to achieve this goal. Here we review the accomplishments in this field by summarizing the history and prospects of microbial isoprene production. The natural producers and biosynthesis pathways of isoprene, the key enzyme isoprene synthase and the metabolic engineering strategies adopted for developing isoprene-producing microorganisms are introduced. In particular, strategies employed for achieving engineered strains with improved performance indices are discussed based on the published papers and patents. The perspectives on further performance improvements and potential future strategies are presented as well.
Assuntos
Alquil e Aril Transferases/metabolismo , Fenômenos Fisiológicos Bacterianos/genética , Fermentação/fisiologia , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/fisiologia , Saccharomyces cerevisiae/fisiologia , Terpenos/metabolismo , Alquil e Aril Transferases/genética , Vias Biossintéticas/fisiologia , Melhoramento Genético/métodos , Análise do Fluxo Metabólico/métodos , Terpenos/isolamento & purificaçãoRESUMO
Metabolic engineering of microorganisms for heterologous biosynthesis is a promising route to sustainable chemical production which attracts increasing research and industrial interest. However, the efficiency of microbial biosynthesis is often restricted by insufficient activity of pathway enzymes and unbalanced utilization of metabolic intermediates. This work presents a combinatorial strategy integrating modification of multiple rate-limiting enzymes and modular pathway engineering to simultaneously improve intra- and inter-pathway balance, which might be applicable for a range of products, using isoprene as an example product. For intra-module engineering within the methylerythritol-phosphate (MEP) pathway, directed co-evolution of DXS/DXR/IDI was performed adopting a lycopene-indicated high-throughput screening method developed herein, leading to 60% improvement of isoprene production. In addition, inter-module engineering between the upstream MEP pathway and the downstream isoprene-forming pathway was conducted via promoter manipulation, which further increased isoprene production by 2.94-fold compared to the recombinant strain with solely protein engineering and 4.7-fold compared to the control strain containing wild-type enzymes. These results demonstrated the potential of pathway optimization in isoprene overproduction as well as the effectiveness of combining metabolic regulation and protein engineering in improvement of microbial biosynthesis. Biotechnol. Bioeng. 2016;113: 2661-2669. © 2016 Wiley Periodicals, Inc.
Assuntos
Vias Biossintéticas/genética , Evolução Molecular Direcionada/métodos , Escherichia coli/fisiologia , Melhoramento Genético/métodos , Hemiterpenos/biossíntese , Engenharia Metabólica/métodos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Butadienos , Técnicas de Química Combinatória/métodos , Hemiterpenos/genética , Taxa de Depuração Metabólica , Pentanos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
Balanced utilization of metabolic intermediates and controllable expression of genes in biosynthetic pathways are key issues for the effective production of value-added chemicals in microbes. An inducer/repressor-free sequential control strategy regulated by glucose concentration in the growth environment was proposed to address these issues, and its efficiency was validated using heterologous beta-carotenoid biosynthesis in Saccharomyces cerevisiae as an example. Through sequential control of the downstream, upstream, and competitive pathways of farnesyl diphosphate (FPP), the crucial metabolic node in the biosynthesis of terpenoids, in a predetermined order, a carotenoid production of 1156 mg/L (20.79 mg/g DCW) was achieved by high-cell density fermentation. Quantitative PCR analysis of the regulated genes demonstrated that the transcription patterns were controlled in a sequential manner as expected. The inducer/repressor-free nature of this strategy offers a both practical and economically efficient approach to improved biosynthetic production of value-added chemicals.
Assuntos
Carotenoides , Engenharia Metabólica , Fosfatos de Poli-Isoprenil/metabolismo , Saccharomyces cerevisiae , Sesquiterpenos/metabolismo , Carotenoides/biossíntese , Carotenoides/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMO
Improved supply of farnesyl diphosphate (FPP) is often considered as a typical strategy for engineering Saccharomyces cerevisiae towards efficient terpenoid production. However, in the engineered strains with enhanced precursor supply, the production of the target metabolite is often impeded by insufficient capacity of the heterologous terpenoid pathways, which limits further conversion of FPP. Here, we tried to assemble an unimpeded biosynthesis pathway by combining directed evolution and metabolic engineering in S. cerevisiae for lycopene-overproduction. First, the catalytic ability of phytoene syntheses from different sources was investigated based on lycopene accumulation. Particularly, the lycopene cyclase function of the bifunctional enzyme CrtYB from Xanthophyllomyces dendrorhous was inactivated by deletion of functional domain and directed evolution to obtain mutants with solely phytoene synthase function. Coexpression of the resulting CrtYB11M mutant along with the CrtE and CrtI genes from X. dendrorhous, and the tHMG1 gene from S. cerevisiae led to production of 4.47 mg/g DCW (Dry cell weight) of lycopene and 25.66 mg/g DCW of the by-product squalene. To further increase the FPP competitiveness of the lycopene synthesis pathway, we tried to enhance the catalytic performance of CrtE by directed evolution and created a series of pathway variants by varying the copy number of Crt genes. Finally, fed-batch fermentation was conducted for the diploid strain YXWPD-14 resulting in accumulation of 1.61 g/L (24.41 mg/g DCW) of lycopene, meanwhile, the by-production of squalene was reduced to below 1 mg/g DCW.
Assuntos
Carotenoides , Evolução Molecular Direcionada/métodos , Engenharia Metabólica/métodos , Saccharomyces cerevisiae , Carotenoides/biossíntese , Carotenoides/genética , Licopeno , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMO
In this report, UbiE and UbiH in the quinone modification pathway (QMP) were identified in addition to UbiG as bottleneck enzymes in the CoQ10 biosynthesis by Rhodobacter sphaeroides. The CoQ10 content was enhanced after co-overexpression of UbiE and UbiG, however, accompanied by the accumulation of the intermediate 10P-MMBQ. UbiH was then co-overexpressed to pull the metabolic flux towards downstream, resulting in an elevated CoQ10 productivity and decreased biomass. On the other hand, the expression levels of UbiE and UbiG were tuned to eliminate the intermediate accumulation, however at the sacrifice of productivity. To alleviate the detrimental effect on either productivity or cell growth, we tried to fuse UbiG with UbiE and localize them onto the membrane to elevate intermediate conversion. By fusing UbiE and UbiG to pufX, CoQ10 was accumulated to 108.51±2.76mg/L with a biomass of 12.2±0.9g/L. At last, we combined the optimized QMP and the previously engineered 2-methyl-d-erythritol-4-phosphate pathway (MEP) to further boost CoQ10 biosynthesis, resulting in a strain with 138±2.64mg/L CoQ10 production.
Assuntos
Proteínas de Bactérias , Metiltransferases , Oxigenases de Função Mista , Oxo-Ácido-Liases , Rhodobacter sphaeroides , Ubiquinona/análogos & derivados , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Metiltransferases/biossíntese , Metiltransferases/genética , Oxigenases de Função Mista/biossíntese , Oxigenases de Função Mista/genética , Oxo-Ácido-Liases/biossíntese , Oxo-Ácido-Liases/genética , Rhodobacter sphaeroides/genética , Rhodobacter sphaeroides/metabolismo , Ubiquinona/biossíntese , Ubiquinona/genéticaRESUMO
Astaxanthin is a highly valued carotenoid with strong antioxidant activity and has wide applications in aquaculture, food, cosmetic, and pharmaceutical industries. The market demand for natural astaxanthin promotes research in metabolic engineering of heterologous hosts for astaxanthin production. In this study, an astaxanthin-producing Saccharomyces cerevisiae strain was created by successively introducing the Haematococcus pluvialis ß-carotenoid hydroxylase (crtZ) and ketolase (bkt) genes into a previously constructed ß-carotene hyperproducer. Further integration of strategies including codon optimization, gene copy number adjustment, and iron cofactor supplementation led to significant increase in the astaxanthin production, reaching up to 4.7 mg/g DCW in the shake-flask cultures which is the highest astaxanthin content in S. cerevisiae reported to date. Besides, the substrate specificity of H. pluvialis CrtZ and BKT and the probable formation route of astaxanthin from ß-carotene in S. cerevisiae were figured out by expressing the genes separately and in combination. The yeast strains engineered in this work provide a basis for further improving biotechnological production of astaxanthin and might offer a useful general approach to the construction of heterologous biosynthetic pathways for other natural products.
Assuntos
Proteínas de Algas/metabolismo , Vias Biossintéticas , Engenharia Metabólica/métodos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Volvocida/enzimologia , Proteínas de Algas/genética , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Oxigenases/genética , Oxigenases/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Volvocida/genética , Xantofilas/biossínteseRESUMO
Methyl (R)-o-chloromandelate (R-CMM) is an intermediate for the platelet aggregation inhibitor clopidogrel. Its preparation through enzymatic resolution of the corresponding ester has been hindered by the lack of an enzyme with satisfying enantioselectivity and activity. In the present work, we aimed to improve the enzymatic enantioselectivity towards methyl (S)-o-chloromandelate (S-CMM) by rational design, using esterase BioH as a model enzyme. Based on the differences in the binding mode of S- and R-enantiomers at the active cavity of the enzyme, the steric and electronic interactions between the key amino acids of BioH and the enantiomers were finely tuned. The enantioselectivity of esterase BioH towards S-CMM was improved from 3.3 (the wild type) to 73.4 (L123V/L181A/L207F). Synergistic interaction was observed between point mutations, and insight into the source of enzymatic enantioselectivity was gained by molecular dynamics simulations. The results can provide a reference for the enzyme design of other enzymes towards S-CMM for the enhancement of enantioselectivity.
Assuntos
Esterases/metabolismo , Ácidos Mandélicos/metabolismo , Estereoisomerismo , Domínio Catalítico , Esterases/química , Esterases/genética , Modelos Moleculares , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Mutação Puntual , Conformação Proteica , Engenharia de Proteínas , Especificidade por SubstratoRESUMO
Saccharomyces cerevisiae is an important platform organism for the synthesis of a great number of natural products. However, the assembly of controllable and genetically stable heterogeneous biosynthetic pathways in S. cerevisiae still remains a significant challenge. Here, we present a strategy for reconstructing controllable multi-gene pathways by employing the GAL regulatory system. A set of marker recyclable integrative plasmids (pMRI) was designed for decentralized assembly of pathways. As proof-of-principle, a controllable ß-carotene biosynthesis pathway (â¼16 kb) was reconstructed and optimized by repeatedly using GAL10-GAL1 bidirectional promoters with high efficiency (80-100%). By controling the switch time of the pathway, production of 11 mg/g DCW of total carotenoids (72.57 mg/L) and 7.41 mg/g DCW of ß-carotene was achieved in shake-flask culture. In addition, the engineered yeast strain exhibited high genetic stability after 20 generations of subculture. The results demonstrated a controllable and genetically stable biosynthetic pathway capable of increasing the yield of target products. Furthermore, the strategy presented in this study could be extended to construct other pathways in S. cerevisisae.
Assuntos
Marcadores Genéticos/genética , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/genética , Plasmídeos/genética , beta Caroteno/biossíntese , Glucose/metabolismo , Glucose/farmacologia , Redes e Vias Metabólicas/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , beta Caroteno/análise , beta Caroteno/genética , beta Caroteno/metabolismoRESUMO
A novel high-throughput screening method is proposed for the directed evolution of exoglucanase facilitated by the co-expression of ß-glucosidase, using the glucose released from filter paper as the screening indicator. Three transformants (B1, D6 and G10) with improved activity were selected from 4,000 colonies. The specific activities of B1, D6 and G10 for releasing glucose were, respectively, 1.4-, 1.3- and 1.6-fold higher than that of the wild type. The engineered exoglucanase gene was inserted into an expression vector carrying the previously engineered endoglucanase and ß-glucosidase genes, and transformed into Escherichia coli to form a completely engineered cellulase system that showed 8.2-fold increase in glucose production (relative activity) compared to the cells equipped with wild-type enzymes. To our knowledge, this is the first report for directed evolution of an exoglucanase using insoluble cellulose as the screening substrate.
Assuntos
Celulase/metabolismo , Evolução Molecular Direcionada , Escherichia coli/enzimologia , Engenharia Metabólica , Celulase/genética , Escherichia coli/genética , Vetores Genéticos , Glucose/metabolismo , Transformação BacterianaRESUMO
Operation performances of phosphorus removal sludge with different electron acceptors in three parallel SBRs were firstly compared in the present study, and the effect of post-aeration on denitrifying phosphorus removal was also studied. Moreover, community dynamics of different phosphorus removal sludge was systematically investigated with high-throughput sequencing for the first time. TP removal rates for nitrate-, nitrite-, and oxygen-based phosphorus removal sludge were 84.8, 78.5, and 87.4 %, with an average effluent TP concentration of 0.758, 0.931, and 0.632 mg/l. The average specific phosphorus release and uptake rates were 20.3, 10.8, and 21.5, and 9.43, 8.68, and 10.8 mgP/(gVSS h), respectively. Moreover, electron utilization efficiency of denitrifying phosphorus removal sludge with nitrate as electron acceptor was higher than nitrite, with P/e(-) were 2.21 and 1.51 mol-P/mol-e(-), respectively. With the assistance of post-aeration for nitrate-based denitrifying phosphorus removal sludge, settling ability could be improved, with SVI decreased from 120 to 80 and 72 ml/g when post-aeration time was 0, 10, and 30 min, respectively. Moreover, further phosphorus removal could be achieved during post-aeration with increased aeration time. However, the anoxic phosphorus uptake was deteriorated, which was likely a result of shifted microbial community structure. Post-aeration of approximately 10 min was proposed for denitrifying phosphorus removal. Nitrate- and nitrite-based denitrifying phosphorus removal sludge exhibited similar community structure. More phosphorus accumulating organisms were enriched under anaerobic-aerobic conditions, while anaerobic-anoxic conditions were favorable for suppressing glycogen-accumulating organisms. Significant differences in pathogenic bacterial community profiles revealed in the current study indicated the potential public health hazards of non-aeration activated sludge system.
Assuntos
Reatores Biológicos , Elétrons , Fósforo/isolamento & purificação , Esgotos/microbiologia , Desnitrificação , Nitratos/metabolismo , Nitritos/metabolismo , Oxidantes/metabolismo , Oxigênio/metabolismo , Fósforo/metabolismoRESUMO
An innovative lateral flow sulphur autotrophic denitrification (LFSAD) reactor was developed in this study; the treatment performance was evaluated and compared with traditional sulphur/limestone autotrophic denitrification (SLAD) reactor. Results showed that nitrite accumulation in the LFSAD reactor was less than 1.0 mg/L during the whole operation. Denitrification rate increased with the increased initial alkalinity and was approaching saturation when initial alkalinity exceeded 2.5 times the theoretical value. Higher influent nitrate concentration could facilitate nitrate removal capacity. In addition, denitrification efficiency could be promoted under an appropriate reflux ratio, and the highest nitrate removal percentage was achieved under reflux ratio of 200%, increased by 23.8% than that without reflux. Running resistance was only about 1/9 of that in SLAD reactor with equal amount of nitrate removed, which was the prominent excellence of the new reactor. In short, this study indicated that the developed reactor was feasible for nitrate removal from waters with lower concentrations, including contaminated surface water, groundwater or secondary effluent of municipal wastewater treatment with fairly low running resistance. The innovation in reactor design in this study may bring forth new ideas of reactor development of sulphur autotrophic denitrification for nitrate-contaminated water treatment.
Assuntos
Reatores Biológicos , Nitratos/metabolismo , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/metabolismo , Carbonato de Cálcio , Desnitrificação , Sulfatos/metabolismo , EnxofreRESUMO
BACKGROUND: Gestational diabetes mellitus (GDM) is considered a risk factor for heart metabolic disorder in future mothers and offspring. Ferroptosis is a new type of programmed cell death, which may participate in the occurrence and development of GDM. OBJECTIVE: This study aims to identify ferroptosis-related genes in GDM by bioinformatics methods and to explore their clinical diagnostic value. METHODS: The dataset GSE103552 was analyzed using the Gene Expression Omnibus (GEO) database to screen for differentially expressed genes (DEGs) in GDM. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and proteinprotein interaction (PPI) network were performed. Gene sets for ferroptosis were retrieved in MSigDB and GSVA gene set analysis was performed on the database. Finally, logistic regression was performed to differentiate between GDM patients and controls to screen for diagnostic markers. RESULTS: A total of 179 DEGs were identified in the expression profile of GDM. GO and KEGG enrichment analysis revealed significant enrichment in the TGF-ß, p53 signaling pathway, platelet activation, glutathione metabolism, sensory perception of taste, and leukocyte and vascular endothelial cell migration regulation. DEGs (n = 107) associated with the ferroptosis gene set were screened by GSVA analysis. The screened DEGs for disease and DEGs for ferroptosis scores were intersected and 35 intersected genes were identified. PPI identified two key genes associated with GDM as CCNB2 and CDK1. Wilcox-test showed low expression of CCNB2 and CDK1 in GDM. The area under the ROC curve (AUC) of the CCNB2 and CDK1 prognostic model was 0.822. CONCLUSION: The genes associated with ferroptosis in GDM were CCNB2 and CDK1, which can be used as valid indicators for the diagnosis of GDM.
RESUMO
Ferroptosis and autophagy are two different cellular processes that have recently been highlighted for their potential roles in the pathogenesis and progression of gestational diabetes (GD). This research sought to uncover the crucial genes tied to ferroptosis and autophagy in GD, further investigating their mechanisms. Differentially expressed genes (DEGs) linked to ferroptosis and autophagy in GD were identified using publicly available data. Pathway enrichment, protein interactions, correlation with immune cell infiltration, and diagnostic value of DEGs were analyzed. HTR-8/SVneo cells were subjected to varying glucose levels to evaluate cell viability and the expression of markers related to ferroptosis and proteins associated with autophagy. Crucial DEGs were validated in vitro. A total of 12 DEGs associated with ferroptosis and autophagy in GD were identified, enriched in the PI3K-AKT signaling pathway. These genes exhibited significant correlations with monocyte infiltration, resting CD4 memory T cells, and follicular helper T cells. They exhibited high diagnostic value for GD (AUC: 0.77-0.97). High glucose treatment inhibited cell viability, induced ferroptosis, and activated autophagy in HTR-8/SVneo cells. Validation confirmed altered expression of SNCA, MTDH, HMGB1, TLR4, SOX2, SESN2, and HMOX1 after glucose treatments. In conclusion, ferroptosis and autophagy may play a role in GD development through key genes (e.g., TLR4, SOX2, SNCA, HMOX1, HMGB1). These genes could serve as promising biomarkers for GD diagnosis.
Assuntos
Diabetes Gestacional , Ferroptose , Proteína HMGB1 , Feminino , Humanos , Gravidez , Diabetes Gestacional/genética , Fosfatidilinositol 3-Quinases , Receptor 4 Toll-Like , Autofagia , Glucose , Sestrinas , Proteínas de Membrana , Proteínas de Ligação a RNARESUMO
The prevention and control of heterocyclic aromatic amines (HAA) formation to mitigate of potential risks to humans, can be achieved by targeting their precursors. In this study, the detailed roles of individual and excess component (20 common α-amino acids, creatine, creatinine, and glucose) on HAA formation in roasted beef patties were examined using UPLC-MS/MS. The results confirmed the reported classical precursors of HAAs. Some components regulated the competitive production of Norharman and Harman. Glycine (Gly) and glucose favored Norharman formation, while cysteine (Cys) and phenylalanine (Phe) for Harman. Serine (Ser) and threonine (Thr) were identified as potential precursors for IQx-type HAAs. Interestingly, methionine (Met), Gly, Thr, Cys, alanine (Ala), and Ser were revealed as more targeted underlying precursors for 1,6-DMIP and 1,5,6-TMIP, and the formation mechanism was inferred. Furthermore, Pro, Leu, His, Ile, Lys and Asp were considered as great inhibitors for HAAs.
Assuntos
Creatina , Glucose , Animais , Bovinos , Humanos , Creatinina , Cromatografia Líquida , Espectrometria de Massa com Cromatografia Líquida , Sequência de Aminoácidos , Espectrometria de Massas em Tandem , Aminas , Aminoácidos , Fragmentos de PeptídeosRESUMO
The favorable inhibitory effect of tea polyphenols on heterocyclic aromatic amines (HAAs) has been confirmed in many past studies. The objective of this study was to investigate the structure-activity relationship of catechins that act as inhibitors of HAA formation in chemical models. Two kinds of quantitative structure-activity relationship models for catechin-inhibiting-HAA were established. We chose two kinds of HAAs including 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), and five catechins including epigallocatechin gallate (EGCG), epicatechin gallate (ECG), epigallocatechin (EGC), epicatechin (EC), and catechin (C). The inhibitory effect of five catechins were in the following order: EGCG > ECG > EGC > C > EC. Thereinto, EGCG and ECG showed dramatically better inhibition on the formation of PhIP and MeIQx, especially EGCG. Further, the mechanisms of catechin-inhibiting-HAA were speculated by correlation analysis. The free radical-scavenging ability was predicted to be the most relevant to the inhibitory effect of ECG, EGC, EC and C on HAAs. Differently, the phenylacetaldehyde-trapping ability might be the more important mechanism of EGCG inhibiting PhIP in chemical model system. This study may bring a broader idea for controlling the formation of HAAs according to the structure of catechins.
RESUMO
Polycyclic aromatic hydrocarbons (PAHs), prominent carcinogens formed during food processing, pose health risks through long-term consumption. This study focuses on 16 priority PAHs in the European Union, investigating their formation during pyrolysis. Glucose, amino acids and fatty acids are important food nutrients. To further explore whether these nutrients in food form PAHs during heating, a single chemical model method was used to heat these nutrients respectively, and GC-MS/MS was used to identify and quantify the obtained components. Glucose is the most basic nutrient in food, so the influence of water, pH, temperature and other factors on the formation of PAHs was studied in the glucose model. At the same time, the models of amino acids and fatty acids were used to assist in improving the entire nutrient research system. According to our results, some previously reported mechanisms of PAHs formation by fatty acids heating were confirmed. In addition, glucose and amino acids could also produce many PAHs after heating, and some conclusions were improved by comparing the intermediates of PAHs from three types of nutrients.