RESUMEN
Micro bunched electron beams with periodic longitudinal density modulation at optical wavelengths give rise to coherent light emission. In this paper, we show attosecond micro bunched beam generation and acceleration in laser-plasma wakefield via particle-in-cell simulations. Due to the near-threshold ionization with the drive laser, the electrons with phase-dependent distributions are non-linearly mapped to discrete final phase spaces. Electrons can preserve this initial bunching structure during the acceleration, leading to an attosecond electron bunch train after leaving the plasma with separations of the same time scale. The modulation of the comb-like current density profile is about 2k0 â¼ 3k0, where k0 is the wavenumber of the laser pulse. Such pre-bunched electrons with low relative energy spread may have potential in applications related to future coherent light sources driven by laser-plasma accelerators and broad application prospects in attosecond science and ultrafast dynamical detection.
RESUMEN
Strain Jing01T, a novel actinomycete from rhizosphere soil of Cathaya argyrophylla, was identified using a polyphasic approach. 16S rRNA gene sequence analysis of strain Jing01T revealed that it was a member of the genus Streptomyces and shared 99.03%, 99.03%, 98.96%, 98.89%, 98.83%, 98.82%, 98.76%, 98.74%, 98.73%, 98.69% and 98.68% similarities to Streptomyces rochei NRRL B-2410T, Streptomyces naganishii NBRC 12892T, Streptomyces rubradiris JCM 4955T, Streptomyces anandii NRRL B-3590T, Streptomyces aurantiogriseus NBRC 12842T, Streptomyces mutabilis NBRC 12800T, Streptomyces rameus LMG 20326T, Streptomyces djakartensis NBRC 15409T, Streptomyces bangladeshensis JCM 14924T, Streptomyces andamanensis KCTC 29502T and Streptomyces tuirus NBRC 15617T, respectively. In phylogenetic trees constructed based on 16S rRNA gene sequences, strain Jing01T generated a separate branch at the middle of the clade, suggesting it could be a potential novel species. In phylogenomic tree, strain Jing01T was related to S. rubradiris JCM 4955T. In phylogenetic trees based on the gene sequences of atpD, gyrB, recA, rpoB and trpB, strain Jing01T was related to S. bangladeshensis JCM 14924T and S. rubradiris JCM 4955T. Whereas, the multilocus sequence analysis distance, average nucleotide identity and DNA-DNA hybridization values between them were much less than the species-level thresholds. This conclusion was further supported by phenotypic and chemotaxonomic analysis. Consequently, strain Jing01T represents a new Streptomyces species, for which the proposed name is Streptomyces argyrophyllae sp. nov. The type strain is Jing01T (= MCCC 1K05707T = JCM 35923T).
Asunto(s)
Rizosfera , Streptomyces , Filogenia , ARN Ribosómico 16S/genética , Streptomyces/genética , ADNRESUMEN
Strain HUAS 13-4T, a novel endophytic actinobacterium, was isolated from the leaves of Cynara scolymus L. collected from Changde City in China and characterized using a polyphasic approach. Based on 16S rRNA gene sequence analysis, strain HUAS 13-4T shared the highest sequence similarities to Streptomyces leeuwenhoekii C34T (98.90%), Streptomyces harenosi PRKS01-65T (98.83%) and Streptomyces glomeratus LMG 19903T (98.76%). Phylogenetic analysis of 16S rRNA gene sequence indicated that strain HUAS 13-4T was clustered together with Streptomyces bluensis ISP 5564T and Streptomyces cavernae SYSU K10008T. Phylogenomic analysis revealed that strain HUAS 13-4T was most closely related to S. glomeratus JCM 9091T. However, the average nucleotide identity and the digital DNA-DNA hybridization values between them were less than 96.7% and 70% cut-off points recommended for delineating species. Based on a comprehensive comparison of the genome sequences and phenotypic characteristics between strain HUAS 13-4T and its relative, strain HUAS 13-4T (= MCCC 1K08364T = JCM 35919T) should evidently represent a novel Streptomyces species, and the name Streptomyces cynarae sp. nov. is proposed.
Asunto(s)
Actinobacteria , Cynara scolymus , Streptomyces , Ácidos Grasos , Fosfolípidos , Cynara scolymus/genética , Análisis de Secuencia de ADN , Filogenia , ARN Ribosómico 16S/genética , Actinobacteria/genética , Composición de Base , ADN , ADN Bacteriano/genética , Técnicas de Tipificación BacterianaRESUMEN
PURPOSE: Evaluation of the efficacy and safety of IL-2 in the treatment of drug-susceptible tuberculosis. METHODS: First, the cases of diagnosed drug-susceptible tuberculosis were randomized into two groups-the control group that received the background regimen of isoniazid, rifampin, pyrazinamide, and ethambutol, and the experimental group that received the background regimen plus IL-2. The efficacy and safety evaluations were performed throughout the therapy process as well as 12 months after the treatment completion. RESULTS: A total of 1151 patients underwent the randomization, among which 539 (96.2%) of the 560 in the experimental group achieved the sputum culture conversion to negative, compared to the 551 (93.2%) of the 591 in the control group, after 2 months of treatment, with significant difference observed between the groups (P = 0.025). Cavity closure after 2 months in the IL-2 (experimental) group was 60/211 (28.4%) compared to 46/248 (18.5%) in the control group, with a significant difference between the groups (P = 0.001). After treatment completion, the proportion of favorable outcomes was 559/560 (99.8%) in the experimental group and 587/591 (99.3%) in the control group, with no significant difference between the groups. Twelve months after treatment completion, relapse occurred in 15/560 (2.6%) in the IL-2 group and 19/591 (3.2%) in the control group, with no significant difference. CONCLUSION: IL-2 may enhance culture conversion and the cavity closure rate in the early treatment phase, although the enhancement may not be significant after treatment completion.
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Tuberculosis Pulmonar , Tuberculosis , Antituberculosos/uso terapéutico , Quimioterapia Combinada , Humanos , Interleucina-2/uso terapéutico , Resultado del Tratamiento , Tuberculosis/tratamiento farmacológico , Tuberculosis Pulmonar/tratamiento farmacológicoRESUMEN
A Gram-stain negative, rod-shaped bacterial, catalase and oxidase positive strain (83-4T) that formed yellow colonies was isolated from human Meibomian gland secretions. Strain 83-4T belongs to the genus Lysobacter according to phylogenetic analysis based on 16S rRNA gene sequences. The DNA G+C content was 67.1 mol%. The circular genome was 2.6 Mb, which contained 2431 protein-coding sequences, 75 pseudogenes, 46 tRNAs, 3 rRNAs and 4 ncRNAs. A bacteriocin cluster and aryl polyene cluster were also found in the genome. The average nucleotide identity value was 79.6% between isolate 83-4T and the closely related type strain Lysobacter tolerans UM1T. The estimated DNA-DNA hybridization value between strain 83-4T and L. tolerans UM1T was 41.6%. Diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were the major polar lipids. Iso-C15:0, iso-C11:0 3-OH, iso-C11:0 and summed feature 9 (iso-C17:1ω9c) were the major fatty acids. Ubiquinone (Q-8) was the only respiratory quinone. Therefore, based on the data of phylogenetic analysis, chemotaxonomical and biochemical analyses, it is concluded that strain 83-4T represents a novel species of the genus Lysobacter with the name of Lysobacter oculi sp. nov. The type strain is 83-4T (= CGMCC 1.13464T = NRBC 113451T).
Asunto(s)
ADN Bacteriano/genética , Lysobacter/clasificación , Lysobacter/genética , Glándulas Tarsales/microbiología , Composición de Base/genética , Cardiolipinas/metabolismo , Humanos , Lysobacter/metabolismo , Fosfatidiletanolaminas/metabolismo , Filogenia , Seudogenes/genética , ARN Ribosómico 16S/genéticaRESUMEN
BACKGROUND: Because people living with HIV (PLWH) are ageing, they will inevitably develop non-communicable diseases (NCDs) and the number of non-HIV medications will increase. Drug-drug interactions(DDIs) will become an ever-increasing issue. However, little is known about this important issue in Chinese PLWH. This study aimed to investigate the prevalence and risk factors of DDIs among PLWH in China. METHODS: Chinese PLWH aged ≥18 years were enrolled prospectively from October 2018 to April 2019 and after informed consent was obtained, they were ask to fill out a questionnaire about comorbidity and co-medications. Potential DDIs were identified using the University of Liverpool HIV Drug Interaction Checker. RESULTS: A total of 1804 questionnaires were included. Antiretroviral drugs (ARVs) that most frequently were prescribed were lamivudine (96.18%), efavirenz(64.64%) and tenofovir(60.62%). 16.96% of the participations reported current co-infection with HIV and14.69% reported NCDs. 263(14.57%) participations reported they had used co-medications in the past six months while 186(10.31%) reported they were taking co-medications. Age≥50 years (p < 0.001), living in developed areas(p < 0.001) and lower CD4 cell count(p = 0.045) were independently associated with the use of co-medications. Potential DDIs were identified in 54 (19.15%) persons using co-medications. Age≥50 [OR = 2.272(1.241-4.158)], PLWH with NCDs[OR = 2.889(1.509-5.532)] and usage of protease inhibitors[OR = 2.538(1.250-5.156)] were independently associated with the potential DDIs. CONCLUSION: The prevalence of the use of co-medications and potential DDIs among Chinese PLWH are low. Older age, NCDs and use of PIs were risk factors for the potential of developing DDIs. With the aging of PLWH, co-medications and DDIs in China warrants more attention.
Asunto(s)
Antirretrovirales/farmacología , Coinfección/tratamiento farmacológico , Infecciones por VIH/tratamiento farmacológico , Polifarmacia , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Antirretrovirales/uso terapéutico , Recuento de Linfocito CD4 , China/epidemiología , Coinfección/epidemiología , Coinfección/inmunología , Comorbilidad , Estudios Transversales , Interacciones Farmacológicas , Femenino , Infecciones por VIH/sangre , Infecciones por VIH/epidemiología , Infecciones por VIH/inmunología , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Estudios Prospectivos , Factores de Riesgo , Encuestas y Cuestionarios/estadística & datos numéricos , Adulto JovenRESUMEN
3-Deoxy-d-arabino-heptulosonate-7-phosphate synthase (DAHPS) is responsible for the biosynthesis of essential aromatic compounds in microorganisms and plants. It plays a crucial role in the regulation of the carbon flow into the shikimate pathway. Until now, the crystal structures and regulatory mechanisms of dimeric DAHPS enzymes from type Iα subclass have not been reported. Here, we reported dimeric structures of the tyrosine-regulated DAHPS from Escherichia coli, both in its apo form and complex with the inhibitor tyrosine at 2.5 and 2.0â¯Å resolutions, respectively. DAHPS(Tyr) has a typical (ß/α)8 TIM barrel, which is decorated with an N-terminal extension and an antiparallel ß sheet, ß6a/ß6b. Inhibitor tyrosine binds at a cavity formed by residues of helices α3, α4, strands ß6a, ß6b and the adjacent loops, and directly interacts with residues P148, Q152, S181, I213 and N8*. Although the small angle X-ray scattering profiles from DAHPS(Tyr) with and without tyrosine shows that tyrosine binding leaves most of DAHPS(Tyr) structures unaffected. The comparison of the liganded and unliganded crystal structures reveals that conformational changes of residues P148, Q152 and I213 initiate a transmission pathway to propagate the allosteric signal from the tyrosine-binding site to the active site, which is different from DAHPS(Phe), a phenylalanine-regulated isozyme from E. coli. In addition, mutations of five tyrosine-binding residues P148, Q152, S181, I213 and N8* leads to tyrosine-resistant DAHPS(Tyr) enzymes. These findings provide a new insight into the regulatory mechanism of DAHPS enzymes and a basis for further engineering studies.
Asunto(s)
3-Desoxi-7-Fosfoheptulonato Sintasa/química , 3-Desoxi-7-Fosfoheptulonato Sintasa/ultraestructura , Escherichia coli/ultraestructura , Conformación Proteica , 3-Desoxi-7-Fosfoheptulonato Sintasa/genética , Regulación Alostérica/genética , Sitios de Unión/genética , Carbono/metabolismo , Dominio Catalítico/genética , Cristalografía por Rayos X , Escherichia coli/química , Escherichia coli/genética , Redes y Vías Metabólicas/genética , Unión Proteica , Estructura Secundaria de Proteína/genética , Ácido Shikímico/metabolismoRESUMEN
Relaxin (RLX) is a pleiotropic peptide hormone with marked renal vasodilatory actions that are physiologically important during pregnancy. RLX also has potent antifibrotic actions and is being tested therapeutically in various fibrotic diseases, including chronic kidney disease (CKD). Since renal vasodilation may expose the glomerulus to increased blood pressure [glomerular capillary pressure (PGC)], which exacerbates progression of CKD, we assessed the glomerular hemodynamic actions of acute (0.89 µg·100 g body wt-1·h-1 iv over 75 min) and chronic (1.5 µg·100 g body wt-1·h-1 sc) administration of RLX. Both acute and chronic RLX produced marked renal vasodilation and increased renal plasma flow (RPF) in euvolemic, anesthetized male rats. Glomerular filtration rate also increased with RLX, but the magnitude of the rise was much less than the increase in RPF due to concomitant decreases in filtration fraction. The fall in filtration fraction was the result of significant decreases in PGC, despite a slight increase in mean arterial blood pressure (MAP) with acute RLX and no net change in MAP with chronic RLX. This fall in PGC occurred because of the "in-series" arrangement of the afferent and efferent arteriolar resistance vessels, which can regulate PGC independently of MAP. With both acute and chronic RLX, efferent arteriolar resistance vessels relaxed to a greater extent than afferent arteriolar resistance vessels, thus producing falls in PGC. Based on this finding, RLX has a beneficial hemodynamic impact on the kidney, which, together with the antifibrotic actions of RLX, suggests a strong therapeutic potential for use in CKD.
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Presión Arterial/efectos de los fármacos , Arteriolas/efectos de los fármacos , Glomérulos Renales/irrigación sanguínea , Relaxina/administración & dosificación , Insuficiencia Renal Crónica/tratamiento farmacológico , Flujo Plasmático Renal/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Vasodilatadores/administración & dosificación , Animales , Arteriolas/fisiopatología , Tasa de Filtración Glomerular/efectos de los fármacos , Infusiones Intravenosas , Masculino , Ratas Sprague-Dawley , Insuficiencia Renal Crónica/fisiopatología , Factores de Tiempo , Resistencia Vascular/efectos de los fármacosRESUMEN
Oxidative stress and inflammation are risk factors for hypertension in pregnancy. Here, we examined the 24-h mean arterial pressure (MAP) via telemetry and the nitric oxide (NO) and redox systems in the kidney cortex, medulla, and aorta of virgin and pregnant rats treated with a high-fat/prooxidant Western diet (HFD), ANG II, and TNF-α. Female Sprague-Dawley rats were given a normal diet (ND) or a HFD for 8 wk before mating. Day 6 of pregnancy and age-matched virgins were implanted with minipumps infusing saline or ANG II (150 ng·kg(-1)·min(-1)) + TNF-α (75 ng/day) for 14 days. Groups consisted of Virgin + ND + Saline (V+ND) (n = 7), Virgin + HFD +ANG II and TNF-α (V+HFD) (n = 7), Pregnant + ND + Saline (P+ND) (n = 6), and Pregnant + HFD + ANG II and TNF-α (P+HFD) (n = 8). After day 6 of minipump implantation, V+HFD rats displayed an increase in MAP on days 7, 8, and 10-15 vs. V+ND rats. P+HFD rats, after day 6 of minipump implantation, showed an increase in MAP only on day 7 vs. P+ND rats. P+HFD rats had a normal fall in 24-h MAP, hematocrit, plasma protein concentration, and osmolality at late pregnancy. No change in kidney cortex, medulla, or aortic oxidative stress in P+HFD rats. P+HFD rats displayed a decrease in nNOSß abundance, but no change in kidney cortex NOx content vs. P+ND rats. Pregnant rats subjected to a chronic HFD and prooxidant and proinflammatory insults have a blunted increase in 24-h MAP and renal oxidative stress. Our data suggest renal NO bioavailability is not altered in pregnant rats treated with a HFD, ANG II, and TNF-α.
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Angiotensina II , Presión Arterial , Dieta Alta en Grasa , Dieta Occidental , Hipertensión/prevención & control , Corteza Renal/metabolismo , Estrés Oxidativo , Factor de Necrosis Tumoral alfa , Animales , Antioxidantes/metabolismo , Aorta/metabolismo , Aorta/fisiopatología , Peso al Nacer , Modelos Animales de Enfermedad , Femenino , Hipertensión/etiología , Hipertensión/metabolismo , Hipertensión/fisiopatología , Tamaño de la Camada , Óxido Nítrico/metabolismo , Embarazo , Ratas Sprague-Dawley , Telemetría , Factores de TiempoRESUMEN
Bacillus subtilis and its closely related species are important strains for industry, agriculture, and medicine. However, it is difficult to perform genetic manipulations using the endogenous recombination machinery. In many bacteria, phage recombineering systems have been employed to improve recombineering frequencies. To date, an efficient phage recombineering system for B. subtilis has not been reported. Here, we, for the first time, identified that GP35 from the native phage SPP1 exhibited a high recombination activity in B. subtilis. On this basis, we developed a high-efficiency GP35-meditated recombineering system. Taking single-stranded DNA (ssDNA) as a recombineering substrate, ten recombinases from diverse sources were investigated in B. subtilis W168. GP35 showed the highest recombineering frequency (1.71 ± 0.15 × 10(-1)). Besides targeting the purine nucleoside phosphorylase gene (deoD), we also demonstrated the utility of GP35 and Beta from Escherichia coli lambda phage by deleting the alpha-amylase gene (amyE) and uracil phosphoribosyltransferase gene (upp). In all three genetic loci, GP35 exhibited a higher frequency than Beta. Moreover, a phylogenetic tree comparing the kinship of different recombinase hosts with B. subtilis was constructed, and the relationship between the recombineering frequency and the kinship of the host was further analyzed. The results suggested that closer kinship to B. subtilis resulted in higher frequency in B. subtilis. In conclusion, the recombinase from native phage or prophage can significantly promote the genetic recombineering frequency in its host, providing an effective genetic tool for constructing genetically engineered strains and investigating bacterial physiology.
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Bacillus subtilis/genética , Bacteriófagos/enzimología , ADN Bacteriano/genética , ADN de Cadena Simple/genética , Ingeniería Genética/métodos , Recombinasas/metabolismo , Recombinación Genética , Proteínas Virales/metabolismo , Bacillus subtilis/metabolismo , Bacteriófagos/genética , ADN Bacteriano/metabolismo , ADN de Cadena Simple/metabolismo , Reacción en Cadena de la Polimerasa , Recombinasas/genética , Proteínas Virales/genéticaRESUMEN
Genetic transformation of bacteria harboring multiple Restriction-Modification (R-M) systems is often difficult using conventional methods. Here, we describe a mimicking-of-DNA-methylation-patterns (MoDMP) pipeline to address this problem in three difficult-to-transform bacterial strains. Twenty-four putative DNA methyltransferases (MTases) from these difficult-to-transform strains were cloned and expressed in an Escherichia coli strain lacking all of the known R-M systems and orphan MTases. Thirteen of these MTases exhibited DNA modification activity in Southwestern dot blot or Liquid Chromatography-Mass Spectrometry (LC-MS) assays. The active MTase genes were assembled into three operons using the Saccharomyces cerevisiae DNA assembler and were co-expressed in the E. coli strain lacking known R-M systems and orphan MTases. Thereafter, results from the dot blot and restriction enzyme digestion assays indicated that the DNA methylation patterns of the difficult-to-transform strains are mimicked in these E. coli hosts. The transformation of the Gram-positive Bacillus amyloliquefaciens TA208 and B. cereus ATCC 10987 strains with the shuttle plasmids prepared from MoDMP hosts showed increased efficiencies (up to four orders of magnitude) compared to those using the plasmids prepared from the E. coli strain lacking known R-M systems and orphan MTases or its parental strain. Additionally, the gene coding for uracil phosphoribosyltransferase (upp) was directly inactivated using non-replicative plasmids prepared from the MoDMP host in B. amyloliquefaciens TA208. Moreover, the Gram-negative chemoautotrophic Nitrobacter hamburgensis strain X14 was transformed and expressed Green Fluorescent Protein (GFP). Finally, the sequence specificities of active MTases were identified by restriction enzyme digestion, making the MoDMP system potentially useful for other strains. The effectiveness of the MoDMP pipeline in different bacterial groups suggests a universal potential. This pipeline could facilitate the functional genomics of the strains that are difficult to transform.
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Bacterias , Metilación de ADN/genética , ADN-Citosina Metilasas/genética , Transformación Genética , Bacillus/genética , Bacterias/genética , Bacterias/metabolismo , Enzimas de Restricción del ADN/genética , Enzimas de Restricción-Modificación del ADN/genética , ADN-Citosina Metilasas/metabolismo , Escherichia coli , Nitrobacter/genética , Saccharomyces cerevisiaeRESUMEN
PDZ domain-containing proteases, also known as HtrA family proteases, play important roles in bacterial cells by modulating disease pathogenesis and cell-envelope stress responses. These proteases have diverse functions through proteolysis- and nonproteolysis-dependent modes. Here, we report that the genome of the causative agent of rice bacterial blight, Xanthomonas oryzae pv. oryzae, encodes seven PDZ domain-containing proteins. Systematic inactivation of their encoding genes revealed that PXO_01122 and PXO_04290 (prc) are involved in virulence. prc encodes a putative HtrA family protease that localizes in the bacterial periplasm. Mutation of prc also resulted in susceptibility to multiple environmental stresses, including H2O2, sodium dodecylsulfate, and osmolarity stresses. Comparative subproteomic analyses showed that the amounts of 34 periplasmic proteins were lower in the prc mutant than in wild-type. These proteins were associated with proteolysis, biosynthesis of macromolecules, carbohydrate or energy metabolism, signal transduction, and protein translocation or folding. We provide in vivo and in vitro evidence demonstrating that Prc stabilizes and directly binds to one of these proteins, DppP, a dipeptidyl peptidase contributing to full virulence. Taken together, our results suggest that Prc contributes to bacterial virulence by acting as a periplasmic modulator of cell-envelope stress responses.
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Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Genoma Bacteriano/genética , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Xanthomonas/fisiología , Proteínas Bacterianas/genética , Peróxido de Hidrógeno/farmacología , Mutación , Presión Osmótica , Dominios PDZ , Péptido Hidrolasas/genética , Péptido Hidrolasas/metabolismo , Periplasma/metabolismo , Fenotipo , Unión Proteica , Mapeo de Interacción de Proteínas , Proteómica , Dodecil Sulfato de Sodio/farmacología , Virulencia , Xanthomonas/efectos de los fármacos , Xanthomonas/genética , Xanthomonas/patogenicidadRESUMEN
At present, it is widely believed that a 95-96% average nucleotide identity (ANI) value is equivalent to a 70% digital DNA-DNA hybridization (dDDH) value in the prokaryotic taxonomy. However, in the present study, comparative genome analysis of 29 pairs of Amycolatopsis type strains revealed that a 70% dDDH value did not correspond to a 95-96% ANI based on the MuMmer ultra-rapid aligning tool (ANIm) but approximately corresponded to a 96.6% ANIm value in the genus Amycolatopsis. Based on this corresponding relationship, phenotypic and chemotaxonomical characteristics, as well as phylogenetic analysis, an actinobacterial strain HUAS 11-8T isolated from the rhizosphere soil of Cynara scolymus, was subjected to a polyphasic taxonomic characterization. Based on EzBioCloud alignment, it was found that strain HUAS11-8T had the 16S rRNA gene similarities of 99.78% with A. rhizosphaerae JCM 32589T, 97.8% with A. dongchuanensis YIM 75904T, and < 97.8% sequence similarities to other Amycolatopsis species. Phylogenetic analysis of 16S rRNA gene sequences and whole-genome sequences revealed that strain HUAS 11-8T was closely related to A. rhizosphaerae JCM 32589T. ANIm and dDDH values between strains HUAS 11-8T and A. rhizosphaerae JCM 32589T were 96.3 and 68.5%, respectively, lower than the 96.6 and 70% thresholds recommended for the delineation of a novel Amycolatopsis species. Consequently, strain HUAS 11-8T should represent a novel Amycolatopsis species, for which the name Amycolatopsis cynarae sp. nov. (type strain HUAS 11-8T = MCCC 1K08337T = JCM 35980T) is proposed. Furthermore, based on comparative genomic analysis and rule 42 of the Prokaryotic Code, we propose that Amycolatopsis niigatensis is a later heterotypic synonym of Amycolatopsis echigonensis.
RESUMEN
Calendula officinalis L.is a versatile medicinal plant with numerous applications in various fields. However, its chloroplast genome structure, features, phylogeny, and patterns of evolution and mutation remain largely unexplored. This study examines the chloroplast genome, phylogeny, codon usage bias, and divergence time of C. officinalis, enhancing our understanding of its evolution and adaptation. The chloroplast genome of C. officinalis is a 150,465 bp circular molecule with a G + C content of 37.75% and comprises 131 genes. Phylogenetic analysis revealed a close relationship between C. officinalis, C. arvensis, and Osteospermum ecklonis. A key finding is the similarity in codon usage bias among these species, which, coupled with the divergence time analysis, supports their close phylogenetic proximity. This similarity in codon preference and divergence times underscores a parallel evolutionary adaptation journey for these species, highlighting the intricate interplay between genetic evolution and environmental adaptation in the Asteraceae family. Moreover unique evolutionary features in C. officinalis, possibly associated with certain genes were identified, laying a foundation for future research into the genetic diversity and medicinal value of C. officinalis.
Asunto(s)
Calendula , Evolución Molecular , Genoma del Cloroplasto , Filogenia , Plantas Medicinales , Plantas Medicinales/genética , Calendula/genética , Uso de Codones , Composición de Base , Cloroplastos/genéticaRESUMEN
Corynebacterium glutamicum is equipped with abundant membrane transporters to adapt to a changing environment. Many amino acid transporters have been identified in C. glutamicum, but histidine uptake has not been investigated in detail. Here, we identified the aromatic amino acid transporter encoded by aroP as a histidine transporter in C. glutamicum by a combination of the growth and histidine uptake features. Characterization of histidine uptake showed that AroP has a moderate affinity for histidine, with a Km value of 11.40 ± 2.03 µM, and histidine uptake by AroP is competitively inhibited by the aromatic amino acids. Among the four substrates, AroP exhibits a stronger preference for tryptophan than for tyrosine, phenylalanine, and histidine. Homology structure modeling and molecular docking were performed to predict the substrate binding modes and conformational changes during substrate transport. These results suggested that tryptophan is best accommodated in the binding pocket due to shape compatibility, strong hydrophobic interactions, and the lowest binding energy, which is consistent with the observed substrate preference of AroP. Furthermore, the missense mutations of the putative substrate binding sites verified that Ser24, Ala28, and Gly29 play crucial roles in substrate binding and are highly conserved in the Gram-positive bacteria. Finally, the expression of aroP is not significantly affected by extracellular histidine or aromatic amino acids, indicating that the physiological role of AroP may be correlated with the increased fitness of C. glutamicum to assimilate extracellular amino acid for avoiding the high energy cost of amino acid biosynthesis.
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Sistemas de Transporte de Aminoácidos/metabolismo , Proteínas Bacterianas/metabolismo , Corynebacterium glutamicum/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Sistemas de Transporte de Aminoácidos/genética , Aminoácidos/metabolismo , Proteínas Bacterianas/genética , Corynebacterium glutamicum/genética , Modelos Moleculares , Filogenia , Unión Proteica , Conformación ProteicaRESUMEN
The rat kidney ablation and infarction (A/I) model of subtotal or 5/6th nephrectomy is the most commonly studied model of nondiabetic chronic kidney disease (CKD). The A/I kidney at 1 wk exhibits reductions in kidney function, as determined by glomerular filtration rate, and diminished metabolic efficiency as determined by oxygen consumption per sodium transport (QO2/TNa). As renoprotective AMPK activity is affected by metabolic changes and cellular stress, we evaluated AMPK activity in this model system. We show that these early pathophysiological changes are accompanied by a paradoxical decrease in AMPK activity. Over time, these kidney parameters progressively worsen with extensive kidney structural, functional, metabolic, and fibrotic changes observed at 4 wk after A/I. We show that induction of AMPK activity with either metformin or 5-aminoimidazole-4-carboxamide ribonucleotide increases AMPK activity in this model and also corrects kidney metabolic inefficiency, improves kidney function, and ameliorates kidney fibrosis and structural alterations. We conclude that AMPK activity is reduced in the subtotal nephrectomy model of nondiabetic CKD, that altered regulation of AMPK is coincident with the progression of disease parameters, and that restoration of AMPK activity can suppress the progressive loss of function characteristic of this model. We propose that induction of AMPK activity may prove an effective therapeutic target for the treatment of nondiabetic CKD.
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Adenilato Quinasa/biosíntesis , Insuficiencia Renal Crónica/fisiopatología , Animales , Modelos Animales de Enfermedad , Inducción Enzimática , Masculino , Metformina/farmacología , Nefrectomía , Ratas , Ratas WistarRESUMEN
Cathaya argyrophylla is an ancient Pinaceae species endemic to China that is listed on the IUCN Red List. Although C. argyrophylla is an ectomycorrhizal plant, the relationship between its rhizospheric soil microbial community and soil properties related to the natural habitat remains unknown. High-throughput sequencing of bacterial 16S rRNA genes and fungal ITS region sequences was used to survey the C. argyrophylla soil community at four natural spatially distributed points in Hunan Province, China, and functional profiles were predicted using PICRUSt2 and FUNGuild. The dominant bacterial phyla included Proteobacteria, Acidobacteria, Actinobacteria, and Chloroflexi, and the dominant genus was Acidothermus. The dominant fungal phyla were Basidiomycota and Ascomycota, while Russula was the dominant genus. Soil properties were the main factors leading to changes in rhizosphere soil bacterial and fungal communities, with nitrogen being the main driver of changes in soil microbial communities. The metabolic capacities of the microbial communities were predicted to identify differences in their functional profiles, including amino acid transport and metabolism, energy production and conversion, and the presence of fungi, including saprotrophs and symbiotrophs. These findings illuminate the soil microbial ecology of C. argyrophylla, and provide a scientific basis for screening rhizosphere microorganisms that are suitable for vegetation restoration and reconstruction for this important threatened species.
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Earth's near space is an extreme atmosphere environment with high levels of radiation, low atmospheric pressure and dramatic temperature fluctuations. The region is above the flight altitude of aircraft but below the orbit of satellites, which has special and Mars-like conditions for investigating the survival and evolution of life. Technical limitations including flight devices, payloads and technologies/methodologies hinder microbiological research in near space. In this study, we investigated microbial survival and adaptive strategies in near space using a scientific balloon fight mission and multi-omics analyses. Methods for sample preparation, storage, protector and vessel were optimized to prepare the exposed microbial samples. After 3 h 17 min of exposure at a float altitude of ~32 km, only Bacillus strains were alive with survival efficiencies of 0-10-6. Diverse mutants with significantly altered metabolites were generated, firstly proving that Earth's near space could be used as a new powerful microbial breeding platform. Multi-omics analyses of mutants revealed cascade changes at the genome, transcriptome and proteome levels. In response to environmental stresses, two mutants had similar proteome changes caused by different genomic mutations and mRNA expression levels. Metabolic network analysis combined with proteins' expression levels revealed that metabolic fluxes of EMP, PPP and purine synthesis-related pathways were significantly altered to increase/decrease inosine production. Further analysis showed that proteins related to translation, molecular chaperones, cell wall/membrane, sporulation, DNA replication/repair and anti-oxidation were significantly upregulated, enabling cells to efficiently repair DNA/protein damages and improve viability against environmental stress. Overall, these results revealed genetic and metabolic responses of Bacillus to the harsh conditions in near space, providing a research basis for bacterial adaptive mechanisms in extreme environments.
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Bacillus , Bacillus/fisiología , Proteoma , Bacterias , Atmósfera , Ambientes ExtremosRESUMEN
BACKGROUND: L-Methionine is the only bulk amino acid that has not been industrially produced by the fermentation method. Due to highly complex and strictly regulated biosynthesis, the development of microbial strains for high-level L-methionine production has remained challenging in recent years. RESULTS: By strengthening the L-methionine terminal synthetic module via site-directed mutation of L-homoserine O-succinyltransferase (MetA) and overexpression of metAfbr, metC, and yjeH, L-methionine production was increased to 1.93 g/L in shake flask fermentation. Deletion of the pykA and pykF genes further improved L-methionine production to 2.51 g/L in shake flask fermentation. Computer simulation and auxotrophic experiments verified that during the synthesis of L-methionine, equimolar amounts of L-isoleucine were accumulated via the elimination reaction of cystathionine γ-synthetase MetB due to the insufficient supply of L-cysteine. To increase the supply of L-cysteine, the L-cysteine synthetic module was strengthened by overexpression of cysEfbr, serAfbr, and cysDN, which further increased the production of L-methionine by 52.9% and significantly reduced the accumulation of the byproduct L-isoleucine by 29.1%. After optimizing the addition of ammonium thiosulfate, the final metabolically engineered strain MET17 produced 21.28 g/L L-methionine in 64 h with glucose as the carbon source in a 5 L fermenter, representing the highest L-methionine titer reported to date. CONCLUSIONS: In this study, a high-efficiency strain for L-methionine production was derived from wild-type Escherichia coli W3110 by rational metabolic engineering strategies, providing an efficient platform for the industrial production of L-methionine.
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BACKGROUND: Artichoke (Cynara scolymus L.) is a typical element of a traditional Mediterranean diet and has potential health advantages for insulin resistance (IR) and type 2 diabetes mellitus (T2DM). This study aims to evaluate the effect and underlying mechanism of artichoke water extract (AWE) on palmitate (PA)-induced IR in human hepatocellular carcinoma (HepG2) cells. METHODS: The effect of AWE on cell viability was determined using CCK8 assay. Cellular glucose uptake, glucose consumption, glucose production, and glycogen content were assessed after AWE treatment. The gene expression and protein levels were examined by real-time polymerase chain reaction (qRT-PCR) and western blotting. RESULTS: The results showed that AWE dose-dependently increased cell viability in IR HepG2 cells (P < 0.01). AWE treatment significantly promoted glucose uptake and consumption, decreased glucose production, and increased the cellular glycogen content in IR HepG2 cells (P < 0.01). Mechanistically, AWE elevated the phosphorylation and total protein levels of major insulin signaling molecules in IR HepG2 cells, which resulted in a decrease in the expression of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) and the inhibition of glycogen synthase (GS) phosphorylation in IR HepG2 cells. Furthermore, the protective effect of AWE on IR HepG2 cells might be ascribed to the inhibition of the endoplasmic reticulum (ER) stress. CONCLUSION: We conclude that AWE may improve glucose metabolism by regulating IRS1/PI3K/AKT/FoxO1 and GSK-3ß signaling associated with the inhibition of ER stress in IR HepG2 cells induced by PA.