RESUMO
Conductive polymers are recognized as ideal candidates for the development of noninvasive and wearable sensors for real-time monitoring of potassium ions (K+) in sweat to ensure the health of life. However, the low ion-to-electron transduction efficiency and limited active surface area hamper the development of high-performance sensors for low-concentration K+ detection in the sweat. Herein, a wearable K+ sensor is developed by tailoring the nanostructure of polypyrrole (PPy), serving as an ion-to-electron transduction layer, for accurately and stably tracing the K+ fluctuation in human sweat. The PPy nanostructures can be tailored from nanospheres to nanofibers by controlling the supramolecular assembly process during PPy polymerization. Resultantly, the ion-to-electron transduction efficiency (17-fold increase in conductivity) and active surface area (1.3-fold enhancement) are significantly enhanced, accompanied by minimized water layer formation. The optimal PPy nanofibers-based K+ sensor achieved a high sensitivity of 62 mV decade-1, good selectivity, and solid stability. After being integrated with a temperature sensor, the manufactured wearable sensor realized accurate monitoring of K+ fluctuation in the human sweat.
Assuntos
Nanofibras , Polímeros , Potássio , Pirróis , Dispositivos Eletrônicos Vestíveis , Nanofibras/química , Pirróis/química , Polímeros/química , Potássio/química , Potássio/análise , Humanos , Técnicas Biossensoriais/métodos , Elétrons , Íons , Suor/química , Condutividade ElétricaRESUMO
The failure of observing the e^{+}e^{-}âJ/ψJ/ψ events at B factories to date is often attributed to the significant negative order-α_{s} correction. In this work we compute the O(α_{s}^{2}) correction to this process for the first time. The magnitude of the next-to-next-to-leading order (NNLO) perturbative correction is substantially negative so that the standard nonrelativistic QCD prediction would suffer from an unphysical, negative cross section. This dilemma may be traced in the fact that the bulk contribution of the fixed-order radiative corrections stems from the perturbative corrections to the J/ψ decay constant. We thus implement an improved nonrelativistic QCD factorization framework, by decomposing the amplitude into the photon-fragmentation piece and the nonfragmentation piece. With the measured J/ψ decay constant as input, which amounts to resumming a specific class of radiative and relativistic corrections to all orders, the fragmentation-induced production rate can be predicted accurately and serves a benchmark prediction. The nonfragmentation type of the amplitude is then computed through NNLO in α_{s} and at lowest order in velocity. Both the O(α_{s}) and O(α_{s}^{2}) corrections in the interference term become positive and exhibit a decent convergence behavior. Our finest prediction is σ(e^{+}e^{-}âJ/ψJ/ψ)=2.13_{-0.06}^{+0.30} fb at sqrt[s]=10.58 GeV. With the projected integrated luminosity of 50 ab^{-1}, the prospect to observe this exclusive process at Belle 2 experiment appears to be bright.
RESUMO
BACKGROUND: Two variants in the gene encoding apolipoprotein L1 (APOL1) that are highly associated with African ancestry are major contributors to the large racial disparity in rates of human kidney disease. We previously demonstrated that recruitment of APOL1 risk variants G1 and G2 from the endoplasmic reticulum to lipid droplets leads to reduced APOL1-mediated cytotoxicity in human podocytes. METHODS: We used CRISPR-Cas9 gene editing of induced pluripotent stem cells to develop human-derived APOL1G0/G0 and APOL1G2/G2 kidney organoids on an isogenic background, and performed bulk RNA sequencing of organoids before and after treatment with IFN-γ. We examined the number and distribution of lipid droplets in response to treatment with inhibitors of diacylglycerol O-acyltransferases 1 and 2 (DGAT1 and DGAT2) in kidney cells and organoids. RESULTS: APOL1 was highly upregulated in response to IFN-γ in human kidney organoids, with greater increases in organoids of high-risk G1 and G2 genotypes compared with wild-type (G0) organoids. RNA sequencing of organoids revealed that high-risk APOL1G2/G2 organoids exhibited downregulation of a number of genes involved in lipogenesis and lipid droplet biogenesis, as well as upregulation of genes involved in fatty acid oxidation. There were fewer lipid droplets in unstimulated high-risk APOL1G2/G2 kidney organoids than in wild-type APOL1G0/G0 organoids. Whereas DGAT1 inhibition reduced kidney organoid lipid droplet number, DGAT2 inhibition unexpectedly increased organoid lipid droplet number. DGAT2 inhibition promoted the recruitment of APOL1 to lipid droplets, with associated reduction in cytotoxicity. CONCLUSIONS: Lipogenesis and lipid droplet formation are important modulators of APOL1-associated cytotoxicity. Inhibition of DGAT2 may offer a potential therapeutic strategy to attenuate cytotoxic effects of APOL1 risk variants.
Assuntos
Nefropatias , Podócitos , Apolipoproteína L1/genética , Diacilglicerol O-Aciltransferase/genética , Feminino , Humanos , Rim , Nefropatias/genética , Gotículas Lipídicas , MasculinoRESUMO
Two coding variants in the apolipoprotein L1 (APOL1) gene (termed G1 and G2) are strongly associated with increased risk of nondiabetic kidney disease in people of recent African ancestry. The mechanisms by which the risk variants cause kidney damage, although not well-understood, are believed to involve injury to glomerular podocytes. The intracellular localization and function of APOL1 in podocytes remain unclear, with recent studies suggesting possible roles in the endoplasmic reticulum (ER), mitochondria, endosomes, lysosomes, and autophagosomes. Here, we demonstrate that APOL1 also localizes to intracellular lipid droplets (LDs). While a large fraction of risk variant APOL1 (G1 and G2) localizes to the ER, a significant proportion of wild-type APOL1 (G0) localizes to LDs. APOL1 transiently interacts with numerous organelles, including the ER, mitochondria, and endosomes. Treatment of cells that promote LD formation with oleic acid shifted the localization of G1 and G2 from the ER to LDs, with accompanying reduction of autophagic flux and cytotoxicity. Coexpression of G0 APOL1 with risk variant APOL1 enabled recruitment of G1 and G2 from the ER to LDs, accompanied by reduced cell death. The ability of G0 APOL1 to recruit risk variant APOL1 to LDs may help explain the recessive pattern of kidney disease inheritance. These studies establish APOL1 as a bona fide LD-associated protein, and reveal that recruitment of risk variant APOL1 to LDs reduces cell toxicity, autophagic flux, and cell death. Thus, interventions that divert APOL1 risk variants to LDs may serve as a novel therapeutic strategy to alleviate their cytotoxic effects.
Assuntos
Apolipoproteína L1/genética , Autofagia/genética , Nefropatias/genética , Gotículas Lipídicas/metabolismo , População Negra/genética , Retículo Endoplasmático/genética , Endossomos/genética , Variação Genética , Células HEK293 , Humanos , Rim/lesões , Rim/patologia , Nefropatias/fisiopatologia , Gotículas Lipídicas/patologia , Lisossomos/genética , Podócitos/metabolismo , Podócitos/patologia , Fatores de RiscoRESUMO
People of recent African ancestry develop kidney disease at much higher rates than most other groups. Two specific coding variants in the Apolipoprotein-L1 gene APOL1 termed G1 and G2 are the causal drivers of much of this difference in risk, following a recessive pattern of inheritance. However, most individuals with a high-risk APOL1 genotype do not develop overt kidney disease, prompting interest in identifying those factors that interact with APOL1 We performed an admixture mapping study to identify genetic modifiers of APOL1-associated kidney disease. Individuals with two APOL1 risk alleles and focal segmental glomerulosclerosis (FSGS) have significantly increased African ancestry at the UBD (also known as FAT10) locus. UBD is a ubiquitin-like protein modifier that targets proteins for proteasomal degradation. African ancestry at the UBD locus correlates with lower levels of UBD expression. In cell-based experiments, the disease-associated APOL1 alleles (known as G1 and G2) lead to increased abundance of UBD mRNA but to decreased levels of UBD protein. UBD gene expression inversely correlates with G1 and G2 APOL1-mediated cell toxicity, as well as with levels of G1 and G2 APOL1 protein in cells. These studies support a model whereby inflammatory stimuli up-regulate both UBD and APOL1, which interact in a functionally important manner. UBD appears to mitigate APOL1-mediated toxicity by targeting it for destruction. Thus, genetically encoded differences in UBD and UBD expression appear to modify the APOL1-associated kidney phenotype.
Assuntos
Apolipoproteína L1/genética , Negro ou Afro-Americano/estatística & dados numéricos , Glomerulosclerose Segmentar e Focal/genética , Glomerulosclerose Segmentar e Focal/patologia , Polimorfismo de Nucleotídeo Único , Ubiquitinas/metabolismo , Alelos , Predisposição Genética para Doença , Genótipo , Glomerulosclerose Segmentar e Focal/etnologia , Humanos , Fatores de Risco , Ubiquitinas/genéticaRESUMO
BACKGROUND: Two coding renal risk variants (RRVs) of the APOL1 gene (G1 and G2) are associated with large increases in CKD rates among populations of recent African descent, but the underlying molecular mechanisms are unknown. Mammalian cell culture models are widely used to study cytotoxicity of RRVs, but results have been contradictory. It remains unclear whether cytotoxicity is RRV-dependent or driven solely by variant-independent overexpression. It is also unknown whether expression of the reference APOL1 allele, the wild-type G0, could prevent cytotoxicity of RRVs. METHODS: We generated tetracycline-inducible APOL1 expression in human embryonic kidney HEK293 cells and examined the effects of increased expression of APOL1 (G0, G1, G2, G0G0, G0G1, or G0G2) on known cytotoxicity phenotypes, including reduced viability, increased swelling, potassium loss, aberrant protein phosphorylation, and dysregulated energy metabolism. Furthermore, whole-genome transcriptome analysis examined deregulated canonical pathways. RESULTS: At moderate expression, RRVs but not G0 caused cytotoxicity in a dose-dependent manner that coexpression of G0 did not reduce. RRVs also have dominant effects on canonical pathways relevant for the cellular stress response. CONCLUSIONS: In HEK293 cells, RRVs exhibit a dominant toxic gain-of-function phenotype that worsens with increasing expression. These observations suggest that high steady-state levels of RRVs may underlie cellular injury in APOL1 nephropathy, and that interventions that reduce RRV expression in kidney compartments may mitigate APOL1 nephropathy.
Assuntos
Apolipoproteína L1/genética , Apolipoproteína L1/fisiologia , Sobrevivência Celular , Metabolismo Energético , Perfilação da Expressão Gênica , Variação Genética , Células HEK293 , Humanos , Potássio/metabolismo , Biossíntese de Proteínas , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologiaRESUMO
BACKGROUND: Genetic Variants in Apolipoprotein L1 (APOL1) are associated with large increases in CKD rates among African Americans. Experiments in cell and mouse models suggest that these risk-related polymorphisms are toxic gain-of-function variants that cause kidney dysfunction, following a recessive mode of inheritance. Recent data in trypanosomes and in human cells indicate that such variants may cause toxicity through their effects on mitochondria. METHODS: To examine the molecular mechanisms underlying APOL1 risk variant-induced mitochondrial dysfunction, we generated tetracycline-inducible HEK293 T-REx cells stably expressing the APOL1 nonrisk G0 variant or APOL1 risk variants. Using these cells, we mapped the molecular pathway from mitochondrial import of APOL1 protein to APOL1-induced cell death with small interfering RNA knockdowns, pharmacologic inhibitors, blue native PAGE, mass spectrometry, and assessment of mitochondrial permeability transition pore function. RESULTS: We found that the APOL1 G0 and risk variant proteins shared the same import pathway into the mitochondrial matrix. Once inside, G0 remained monomeric, whereas risk variant proteins were prone to forming higher-order oligomers. Both nonrisk G0 and risk variant proteins bound components of the mitochondrial permeability transition pore, but only risk variant proteins activated pore opening. Blocking mitochondrial import of APOL1 risk variants largely eliminated oligomer formation and also rescued toxicity. CONCLUSIONS: Our study illuminates important differences in the molecular behavior of APOL1 nonrisk and risk variants, and our observations suggest a mechanism that may explain the very different functional effects of these variants, despite the lack of consistently observed differences in trafficking patterns, intracellular localization, or binding partners. Variant-dependent differences in oligomerization pattern may underlie APOL1's recessive, gain-of-function biology.
Assuntos
Apolipoproteína L1/genética , Falência Renal Crônica/genética , Proteínas de Transporte da Membrana Mitocondrial/fisiologia , Apolipoproteína L1/antagonistas & inibidores , Apolipoproteína L1/fisiologia , Morte Celular , Respiração Celular , Mutação com Ganho de Função , Predisposição Genética para Doença , Células HEK293 , Humanos , Falência Renal Crônica/etnologia , Falência Renal Crônica/microbiologia , Poro de Transição de Permeabilidade Mitocondrial , Multimerização Proteica , Transporte Proteico , Interferência de RNA , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Proteínas Recombinantes/metabolismoRESUMO
Two specific genetic variants of the apolipoprotein L1 (APOL1) gene are responsible for the high rate of kidney disease in people of recent African ancestry. Expression in cultured cells of these APOL1 risk variants, commonly referred to as G1 and G2, results in significant cytotoxicity. The underlying mechanism of this cytotoxicity is poorly understood. We hypothesized that this cytotoxicity is mediated by APOL1 risk variant-induced dysregulation of intracellular signaling relevant for cell survival. To test this hypothesis, we conditionally expressed WT human APOL1 (G0), the APOL1 G1 variant, or the APOL1 G2 variant in human embryonic kidney cells (T-REx-293) using a tetracycline-mediated (Tet-On) system. We found that expression of either G1 or G2 APOL1 variants increased apparent cell swelling and cell death compared with G0-expressing cells. These manifestations of cytotoxicity were preceded by G1 or G2 APOL1-induced net efflux of intracellular potassium as measured by X-ray fluorescence, resulting in the activation of stress-activated protein kinases (SAPKs), p38 MAPK, and JNK. Prevention of net K(+) efflux inhibited activation of these SAPKs by APOL1 G1 or G2. Furthermore, inhibition of SAPK signaling and inhibition of net K(+) efflux abrogated cytotoxicity associated with expression of APOL1 risk variants. These findings in cell culture raise the possibility that nephrotoxicity of APOL1 risk variants may be mediated by APOL1 risk variant-induced net loss of intracellular K(+) and subsequent induction of stress-activated protein kinase pathways.
Assuntos
Apolipoproteínas/genética , Transporte de Íons/genética , Nefropatias/genética , Lipoproteínas HDL/genética , Proteínas Quinases Ativadas por Mitógeno/fisiologia , Mutação de Sentido Incorreto , Potássio/metabolismo , Substituição de Aminoácidos , Apolipoproteína L1 , Apolipoproteínas/fisiologia , População Negra/genética , Morte Celular , Tamanho Celular , Receptor gp130 de Citocina/biossíntese , Receptor gp130 de Citocina/genética , Progressão da Doença , Ativação Enzimática , Frequência do Gene , Predisposição Genética para Doença , Células HEK293 , Humanos , Nefropatias/etnologia , Lipoproteínas HDL/fisiologia , Sistema de Sinalização das MAP Quinases , Fosforilação , Processamento de Proteína Pós-Traducional , Proteínas Recombinantes de Fusão/metabolismo , Risco , Fator de Transcrição STAT3/metabolismo , TransfecçãoRESUMO
Phenolic compounds determine the color quality of fruit wines. In this study, the phenolic compound content and composition, color characteristics and changes during 6 months of bottle aging were studied in wines fermented with bog bilberry syrup under three different pHs. The total anthocyanins and total phenols were around 15.12-16.23 mg/L and 475.82 to 486.50 mg GAE/L in fresh wines and declined 22%-31% and about 11% in bottle aged wines, respectively. In fresh wines, eight anthocyanins, six phenolic aids and 14 flavonols, but no flavon-3-ols were identified; Malvidin-3-O-glucoside, petunidin-3-O-glucoside and delphinium-3-O-glucoside were the predominant pigments; Chlorogentic acid was the most abundant phenolic acid, and quercetin-3-O-galactoside and myricetin-3-O-galactoside accounted for nearly 90% of the total flavonols. During 6 months of bottle storage, the amounts of all the monomeric anthocyanins and phenolic acids were reduced dramatically, while the glycosidyl flavonols remained constant or were less reduced and their corresponding aglycones increased a lot. The effects of aging on blueberry wine color were described as the loss of color intensity with a dramatic change in color hue, from initial red-purple up to final red-brick nuances, while the pH of the fermentation matrix was negatively related to the color stability of aged wine.
Assuntos
Polifenóis/química , Vaccinium myrtillus/química , Vinho/análise , Antocianinas/química , Fermentação , Flavonóis/química , Concentração de Íons de Hidrogênio , Hidroxibenzoatos/química , Fenóis/química , Pigmentos Biológicos/análiseRESUMO
BACKGROUND: Mild cognitive impairment (MCI) is the stage between cognitive decline due to physiological aging and the severity of decline seen in neurodegenerative disorders like Alzheimer disease (AD), which is among the most prevalent neurodegenerative disorders characterized by cognitive impairment. People with MCI are at increased risk of developing AD. Although MCI and AD are incurable, nutritional interventions can potentially delay or prevent their onset. Consequently, effective interventions used to decelerate or alleviate the progress of cognitive impairment in older people are a significant focus in geriatric care. Given the synergistic effects of nutrition on health, assessing the effectiveness of nutritional supplements or dietary composition in preventing MCI or AD is essential for developing interventional strategies. OBJECTIVE: Our study aims to assess the effectiveness of various nutritional interventions, including special dietary types, dietary patterns, specific foods, nutritional intake, and nutritional supplements, in preventing cognitive decline among patients diagnosed with MCI or AD. To achieve this, we will use a comprehensive approach, including network meta-analysis, pairwise meta-analysis, and systematic review of randomized controlled trials (RCTs). METHODS: The review will follow the Population, Intervention, Comparison, Outcome (PICO) model and the PRISMA-P (Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols) guidelines. Two investigators will independently search PubMed electronically. Data extraction will follow the inclusion criteria, and data will be assessed for risk of bias using a revised tool. Additionally, evidence quality will be evaluated using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) framework. The outcomes of interest are assessing the cognitive outcomes in patients with MCI or AD. A systematic literature search will be conducted, identifying randomized controlled trials that investigate the impact of these nutritional interventions on cognitive function decline in individuals with MCI and AD. Network meta-analyses (random-effects model) and pairwise meta-analyses will then estimate the relative effectiveness of different nutritional interventions. RESULTS: We included 51 studies, published between 1999 and 2023 (27 studies for AD and 24 studies for MCI) and involving 8420 participants. We completed data extraction for all 51 studies by December 2023. Currently, we are actively engaged in data analysis and manuscript preparation. We plan to finalize the manuscript and publish the comprehensive results by the end of 2024. CONCLUSIONS: Our study holds significant clinical relevance given the rising prevalence of AD and the potential influence of nutritional interventions on cognitive function in individuals with MCI and AD. By investigating this relationship, our research aims to inform evidence-based decision-making in the development of prevention strategies for MCI and AD. The outcomes are expected to contribute to the establishment of reliable recommendations for MCI or AD management, providing substantial support in the field. TRIAL REGISTRATION: PROSPERO CRD42022331173; http://tinyurl.com/3snjp7a4. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/47196.
RESUMO
Antibiotics easily remain in sediments after migrating from the surface to the subsurface due to water-rock interactions, posing a risk of secondary release to groundwater. To investigate the vertical distribution characteristics and environmental impact factors of antibiotics, five 30 m quaternary sediment columns were drilled and stratified near the hospital, and five major classes of antibiotics and sulfonamide metabolites were tested and analyzed. The results showed that:â the antibiotic content in the sediments ranged from 3.05 to 107.03 µg·kg-1, and all of the target antibiotics were detected except lomefloxacin, of which ofloxacin and oxytetracycline were the most important antibiotics in the study area. â¡ The antibiotics did not show a strict downward trend in the vertical direction but varied with the lithological stratification. ⢠Antibiotics were primarily deposited in the clay layer and varied with the fluctuation of the groundwater level. ⣠The results of redundancy analysis between antibiotics and environmental factors suggested that pH and TOC controlled the fate and transformation of antibiotics through influencing the adsorption of antibiotics by sediments. The risk of antibiotic contamination from hospital wastewater seepage into the subsurface environment should be taken seriously.
Assuntos
Água Subterrânea , Poluentes Químicos da Água , Antibacterianos/análise , Poluentes Químicos da Água/análise , Monitoramento Ambiental , Sedimentos Geológicos/química , ChinaRESUMO
Dioxygen activation by nonhaem Fe(II) enzymes containing the 2-His-1-carboxylate facial triad has been extensively studied in recent years. Here, crystal structures of 2-aminophenol 1,6-dioxygenase, an enzyme that represents a minor group of extradiol dioxygenases and that catalyses the ring opening of 2-aminophenol, in complex with the lactone intermediate (4Z,6Z)-3-iminooxepin-2(3H)-one and the product 2-aminomuconic 6-semialdehyde and in complex with the suicide inhibitor 4-nitrocatechol are reported. The Fe-ligand binding schemes observed in these structures revealed some common geometrical characteristics that are shared by the published structures of extradiol dioxygenases, suggesting that enzymes that catalyse the oxidation of noncatecholic compounds are very likely to utilize a similar strategy for dioxygen activation and the fission of aromatic rings as the canonical mechanism. The Fe-ligation arrangement, however, is strikingly enantiomeric to that of all other 2-His-1-carboxylate enzymes apart from protocatechuate 4,5-dioxygenase. This structural variance leads to the generation of an uncommon O(-)-Fe(2+)-O(-) species prior to O(2) binding, which probably forms the structural basis on which APD distinguishes its specific substrate and inhibitor, which share an analogous molecular structure.
Assuntos
Catecóis/química , Catecóis/farmacologia , Dioxigenases/antagonistas & inibidores , Dioxigenases/química , Aminofenóis/química , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Domínio Catalítico , Comamonas/enzimologia , Cristalização , Cristalografia por Raios X/métodos , Dioxigenases/metabolismo , Evolução Molecular , Ferro/química , Deficiências de Ferro , Subunidades Proteicas/química , Especificidade por SubstratoRESUMO
OBJECTIVE: To explore the characteristics of acute exacerbations of myasthenia gravis after fluoroquinolone exposure. METHODS: Gender, age, prior type, absolute score, concurrent disease, precipitated disease, use of antibiotic, onset/symptom/degree of exacerbation, therapeutic measures and prognosis at Month 1 were retrospectively analyzed for 9 patients after fluoroquinolone systemic exposure. RESULTS: Ciprofloxacin (n = 4), levofloxacin (n = 1) and moxifloxacin (n = 4) exposure resulted in myasthenia gravis exacerbation. Myasthenia gravis exacerbations developed at 15 minutes to 4 days post-exposure. And the clinical scores of quantitative myasthenia gravis (QMG) increased by an average of 10. The main syndromes included dyspnea, diplopia, ptosis and dysphagia. All patients improved upon the withdrawal of fluoroquinolone in conjunctions with other interventions. CONCLUSION: Fluoroquinolone exposure may result in myasthenia gravis exacerbations in patients with underlying diseases. Healthcare professionals should be aware of this serious drug-disease association.
Assuntos
Antibacterianos/efeitos adversos , Fluoroquinolonas/efeitos adversos , Miastenia Gravis/induzido quimicamente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/uso terapêutico , Compostos Aza/efeitos adversos , Compostos Aza/uso terapêutico , Ciprofloxacina/efeitos adversos , Ciprofloxacina/uso terapêutico , Feminino , Fluoroquinolonas/uso terapêutico , Humanos , Levofloxacino/efeitos adversos , Levofloxacino/uso terapêutico , Masculino , Pessoa de Meia-Idade , Moxifloxacina , Miastenia Gravis/tratamento farmacológico , Quinolinas/efeitos adversos , Quinolinas/uso terapêutico , Estudos Retrospectivos , Adulto JovemRESUMO
Dioxygen activation implemented by nonhaem Fe(II) enzymes containing the 2-His-1-carboxylate facial triad has been extensively studied in recent years. Extradiol dioxygenase is the archetypal member of this superfamily and catalyzes the oxygenolytic ring opening of catechol analogues. Here, the crystallization and preliminary X-ray analysis of 2-aminophenol 1,6-dioxygenase, an enzyme representing a minor subset of extradiol dioxygenases that catalyze the fission of 2-aminophenol rather than catecholic compounds, is reported. Crystals of the holoenzyme with FeII and of complexes with the substrate 2-aminophenol and the suicide inhibitor 4-nitrocatechol were grown using the cocrystallization method under the same conditions as used for the crystallization of the apoenzyme. The crystals belonged to space group C2 and diffracted to 2.3-2.7â Å resolution; the crystal that diffracted to the highest resolution had unit-cell parameters a=270.24, b=48.39, c=108.55â Å, ß=109.57°. All X-ray data sets collected from diffraction-quality crystals were suitable for structure determination.
Assuntos
Aminofenóis/química , Proteínas de Bactérias/química , Catecóis/química , Comamonas testosteroni/enzimologia , Dioxigenases/química , Apoenzimas/química , Apoenzimas/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Cromatografia em Gel , Cromatografia por Troca Iônica , Cristalização , Cristalografia por Raios X , Dioxigenases/isolamento & purificação , Inibidores Enzimáticos/químicaRESUMO
A bacterial strain, designated VA24(T), was isolated from forest soil of the Changbai Mountains, Heilongjiang province, China. Cells of strain VA24(T) were Gram-reaction-negative, aerobic, non-spore-forming, long rods, 0.3-0.5×2.0-3.0 µm in size and were motile by means of a subpolar flagellum. Strain VA24(T) was oxidase-positive and catalase-negative. Growth occurred at 21-36 °C, pH 5-10 and in 0-2â% (w/v) NaCl but did not occur at 37 °C. The predominant respiratory quinone was Q-8, the major polar lipids were phosphatidylethanolamine and phosphatidylmonomethylethanolamine and the major cellular fatty acids were iso-C(15â:â0) (14.9â%), iso-C(17â:â1)ω9c (14.1â%), iso-C(17â:â0) (10.8â%) and iso-C(16â:â0) (10.3â%). The DNA G+C content was 67.4 mol% (T(m)). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain VA24(T) was closely related to Frateuria aurantia IFO 13300(T) with 96.9â% sequence similarity. DNA-DNA relatedness of strain VA24(T) to F. aurantia DSM 6220(T) was 15.8â%. Based on its phenotypic and genotypic features, strain VA24(T) represents a novel species of the genus Frateuria, for which the name Frateuria terrea sp. nov. is proposed. The type strain is VA24(T) (=CGMCC 1.7053(T) =NBRC 104236(T)).
Assuntos
Filogenia , Microbiologia do Solo , Árvores/microbiologia , Xanthomonadaceae/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/análise , Flagelos/microbiologia , Genótipo , Dados de Sequência Molecular , Fenótipo , Fosfatidiletanolaminas/análise , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Xanthomonadaceae/genética , Xanthomonadaceae/isolamento & purificaçãoRESUMO
A bacterial strain, designated V2M29(T), was isolated from forest soil collected in the Changbai Mountains, Heilongjiang Province, China. Cells of strain V2M29(T) were Gram-positive, rod-shaped (0.5-0.8 µm in diameter and 1.5-2.0 µm in length), strictly aerobic and non-motile. Colonies produced on R2A agar plates were creamy, smooth, circular and 0.3-0.8 mm in diameter. Strain V2M29(T) grew at 14-35 °C (optimum, 29 °C), at pH 4.1-10.0 (optimum, pH 7.0-8.0) and in the presence of 0-7.0â% (w/v) NaCl (optimum, 0-2.0â%). Strain V2M29(T) contained MK-8(H(4)) as the major respiratory quinone and iso-C(15â:â0) (20.9â%), iso-C(16â:â0) (18.5â%), C(17â:â1)ω8c (16.7â%) and 10-methyl-C(17â:â0) (9.2â%) as the major cellular fatty acids. The cell-wall peptidoglycan type was based on meso-2,6-diaminopimelic acid and the acyl type of the muramic acid was acetyl. Diphosphatidylglycerol, phosphatidylinositol and six unknown glycolipids were the major polar lipids. The G+C content of the DNA was 72.0 mol% (T(m)). 16S rRNA gene sequence analysis indicated that strain V2M29(T) was phylogenetically related to members of the genus Phycicoccus, with sequence similarities ranging from 96.6-97.8â%. The DNA-DNA relatedness values of strain V2M29(T) to Phycicoccus dokdonensis DS-8(T) and Phycicoccus bigeumensis DSM 19264(T) were 32.5±3.3â% and 29.2±2.3â%, respectively. Based on these results, it is concluded that strain V2M29(T) represents a novel species of the genus Phycicoccus, for which the name Phycicoccus cremeus sp. nov. is proposed, with V2M29(T) (=CGMCC 1.6963(T) =NBRC 104261(T)) as the type strain.
Assuntos
Actinomycetales/classificação , Actinomycetales/isolamento & purificação , Microbiologia do Solo , Actinomycetales/genética , Actinomycetales/fisiologia , Aerobiose , Técnicas Bacteriológicas , Composição de Bases , China , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácido Diaminopimélico/análise , Ácidos Graxos/análise , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Peptidoglicano/química , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , Temperatura , ÁrvoresRESUMO
Potato (Solanum tuberosum L.), a species of the family Solanaceae, is the fourth most important food crop worldwide. Solanum tuberosum L. cv. Shepody is a long, smooth, white-skinned potato cultivar with medium green leaves. It has good specific gravity and boils and bakes well. To support more molecular data for breeding of S. tuberosum, the complete chloroplast (cp) genome sequence of S. tuberosum L. cv. Shepody was determined using the next-generation sequencing. In leaves, the chloroplast genome accounts for 3.88% of the total genome. The entire cp genome was determined to be 155,296 bp in length. It contained large single-copy (LSC) and small single-copy (SSC) regions of 85,737 and 18,373 bp, respectively, which were separated by a pair of 25,593 bp inverted repeat (IR) regions. The genome contained 132 total genes, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content of the genome is 37.9%. A phylogenetic tree reconstructed by 60 chloroplast genomes reveals that S. tuberosum L. cv. Shepody was closely related to S. tuberosum L. cv. Desiree with bootstrap support values of 100%.
RESUMO
Potato (Solanum tuberosum L.), a species of the family Solanaceae, is the fourth most important food crop worldwide. Solanum tuberosum L. cv. Favorita is a long oval, smooth, yellowish-skinned potato variety with green and plump leaves. It has a dry matter content of 17.7% and starch content of 12.4-14.01% in the tuber. In order to support more genetic data for the taxonomy of S. tuberosum, the complete chloroplast (cp) genome sequence of S. tuberosum L. cv. Favorita was determined using next-generation sequencing. In leaves, the chloroplast genome accounts for 5.17% of the total genome. The entire cp genome was determined to be 155,296 bp in length. It contained large single-copy (LSC) and small single-copy (SSC) regions of 85,737 and 18,373 bp, respectively, which were separated by a pair of 25,593 bp inverted repeat (IR) regions. The genome contained 132 total genes, including 87 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The overall GC content of the genome is 37.9%. A phylogenetic tree reconstructed by 60 chloroplast genomes reveals that S. tuberosum L. cv. Favorita is most closely related to S. tuberosum L. cv. Desiree and S. tuberosum L. cv. Atlantic.
RESUMO
Potato (Solanum tuberosum L.), a species of the family Solanaceae, is the fourth most important food crop worldwide. Solanum tuberosum L. cv. Atlantic, a main fried special potato, has a dry matter content of 19%-23% and a starch content of 16.26% in the tuber. In order to support more molecular data for the taxony of S. tuberosum, the complete chloroplast (cp) genome sequence of S. tuberosum L. cv. Atlantic was determined using next-generation sequencing. In leaves, the chloroplast genome accounts for 5.49% of the total genome. The entire cp genome was determined to be 155,296 bp in length. It contained large single-copy (LSC) and small single-copy (SSC) regions of 85,737 and 18,373 bp, respectively, which were separated by a pair of 25,593 bp inverted repeat (IR) regions. The genome contained 132 genes, including 87 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The overall GC content of the genome is 37.9%. A phylogenetic tree reconstructed by 64 chloroplast genomes reveals that S. tuberosum L. cv. Atlantic is most closely related to Solanum tuberosum L. cv. Desiree.
RESUMO
The Japanese threadfin bream N. japonicus (Bloch) (Perciformes: Nemipteridae) is an important marine food fish in Asia. However, our present knowledge of the occurrence of its nematode parasites is still limited. In the present study, the species composition and infection rate of ascaridoid nematodes in N. japonicus from the South China Sea, were studied for the first time. Five ascaridoid species, namely Anisakis typica (L3), Hysterothylacium amoyense (L3), Hysterothylacium sp. IV-A (L3), adult of H. thalassini and Raphidascaris lophii (L3), were identified using integrative taxonomy. Hysterothylacium amoyense was the most prevalent species (prevalence 47.2%, mean intensity 14.9 ± 17.1). Hysterothylacium thalassini and R. lophii were reported in the Japanese threadfin bream for the first time. Two different genotypes of A. typica (overall prevalence of 3.4%; mean intensity 1.7 ± 0.9) were found in the South China Sea for the first time. The unique restriction polymorphism patterns of three species of Hysterothylacium are provided for rapid diagnosis. Our present results indicate that RFLP analysis of ITS region, using the restriction enzymes HhaI and RsaI, represents a simple and practical method for large-scale surveys of Hysterothylacium for seafood industry.