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1.
J Antimicrob Chemother ; 76(6): 1459-1466, 2021 05 12.
Artigo em Inglês | MEDLINE | ID: mdl-33704480

RESUMO

BACKGROUND: There are approximately 300 cases of enteric fever reported annually from England and Wales; most are imported infections. Clinical management of enteric fever remains a challenge with the emergence of ESBL-producing strains, especially XDR Salmonella Typhi from Sindh, Pakistan. METHODS: All strains of S. Typhi and Salmonella Paratyphi A isolated from cases presenting with symptoms of enteric fever in England and Wales, between 1 April 2014 and 31 March 2020, were characterized using WGS. Antibiotic susceptibility testing was performed using an agar dilution method. RESULTS: ESBL strains contributed to 69 cases of enteric fever (S. Typhi n = 68, S. Paratyphi A n = 1); 68 were imported (Pakistan n = 64, Iraq n = 2, Bangladesh n = 1 and India n = 1). Ages ranged from 1 to 56 years, 36/69 (52%) were children, 52% were female and the duration of hospital stay ranged from 1 to 23 days. The ESBL phenotype was conferred by the presence of blaCTX-M-15 (S. Typhi n = 67 and S. Paratyphi A n = 1) or blaCTX-M-55 (S. Typhi n = 1). An IncY plasmid harbouring blaCTX-M-15 and qnr was detected in 56 strains from Pakistan. The IncY plasmid was absent in the remaining strains and there was evidence of a 4 kb ISEcpl-blaCTX-M-15-tnp gene cassette insertion into the chromosome at one of three integration points. CONCLUSIONS: Chromosomal integration of blaCTX-M-15 within the XDR Sindh strains may lead to the maintenance of resistance in the absence of antibiotic selection pressure. Empirical treatment of cases of complicated enteric fever returning from Pakistan will henceforth have to include a carbapenem.


Assuntos
Salmonella typhi , Febre Tifoide , Adolescente , Adulto , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bangladesh , Criança , Pré-Escolar , Cromossomos , Inglaterra/epidemiologia , Feminino , Humanos , Índia , Lactente , Pessoa de Meia-Idade , Paquistão , Salmonella typhi/genética , Febre Tifoide/epidemiologia , País de Gales/epidemiologia , Adulto Jovem , beta-Lactamases/genética
2.
Faraday Discuss ; 215(0): 26-38, 2019 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-30969289

RESUMO

The high quantum efficiency of natural photosynthesis has inspired chemists for solar fuel synthesis. In photosynthesis, charge recombination in photosystems is minimized by efficient charge separation across the thylakoid membrane. Building on our previous bioelectrochemical studies of electron transfer between a light-harvesting nanoparticle (LHNP) and the decahaem subunit MtrC, we demonstrate photo-induced electron transfer through the full transmembrane MtrCAB complex in liposome membranes. Successful photoelectron transfer is demonstrated by the decomposition of a redox dye, Reactive Red 120 (RR120), encapsulated in MtrCAB proteoliposomes. The photoreduction rates are found to be dependent on the identity of the external LHNPs, specifically, dye-sensitized TiO2, amorphous carbon dots (a-CD) and graphitic carbon dots with core nitrogen doping (g-N-CDs). Agglomeration or aggregation of TiO2 NPs likely reduces the kinetics of RR120 reductive decomposition. In contrast, with the dispersed a-CD and g-N-CDs, the kinetics of the RR120 reductive decomposition are observed to be faster with the MtrCAB proteoliposomes and we propose that this is due to enhancement in the charge-separated state. Thus, we show a proof-of-concept for using MtrCAB as a lipid membrane-spanning building block for compartmentalised photocatalysis that mimics photosynthesis. Future work is focused on incorporation of fuel generating redox catalysts in the MtrCAB proteoliposome lumen.


Assuntos
Proteínas de Bactérias/metabolismo , Fotossíntese , Proteínas de Bactérias/química , Biocatálise , Transporte de Elétrons , Shewanella/química , Shewanella/metabolismo
3.
Chembiochem ; 17(24): 2324-2333, 2016 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-27685371

RESUMO

The transfer of photoenergized electrons from extracellular photosensitizers across a bacterial cell envelope to drive intracellular chemical transformations represents an attractive way to harness nature's catalytic machinery for solar-assisted chemical synthesis. In Shewanella oneidensis MR-1 (MR-1), trans-outer-membrane electron transfer is performed by the extracellular cytochromes MtrC and OmcA acting together with the outer-membrane-spanning porin⋅cytochrome complex (MtrAB). Here we demonstrate photoreduction of solutions of MtrC, OmcA, and the MtrCAB complex by soluble photosensitizers: namely, eosin Y, fluorescein, proflavine, flavin, and adenine dinucleotide, as well as by riboflavin and flavin mononucleotide, two compounds secreted by MR-1. We show photoreduction of MtrC and OmcA adsorbed on RuII -dye-sensitized TiO2 nanoparticles and that these protein-coated particles perform photocatalytic reduction of solutions of MtrC, OmcA, and MtrCAB. These findings provide a framework for informed development of strategies for using the outer-membrane-associated cytochromes of MR-1 for solar-driven microbial synthesis in natural and engineered bacteria.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Corantes/química , Grupo dos Citocromos c/metabolismo , Titânio/química , Catálise , Transporte de Elétrons , Amarelo de Eosina-(YS)/química , Compostos Férricos/química , Mononucleotídeo de Flavina/química , Luz , Nanopartículas Metálicas/química , Oxirredução , Fármacos Fotossensibilizantes/química , Shewanella
4.
Adv Funct Mater ; 25(15): 2308-2315, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26180522

RESUMO

In nature, charge recombination in light-harvesting reaction centers is minimized by efficient charge separation. Here, it is aimed to mimic this by coupling dye-sensitized TiO2 nanocrystals to a decaheme protein, MtrC from Shewanella oneidensis MR-1, where the 10 hemes of MtrC form a ≈7-nm-long molecular wire between the TiO2 and the underlying electrode. The system is assembled by forming a densely packed MtrC film on an ultra-flat gold electrode, followed by the adsorption of approximately 7 nm TiO2 nanocrystals that are modified with a phosphonated bipyridine Ru(II) dye (RuP). The step-by-step construction of the MtrC/TiO2 system is monitored with (photo)electrochemistry, quartz-crystal microbalance with dissipation (QCM-D), and atomic force microscopy (AFM). Photocurrents are dependent on the redox state of the MtrC, confirming that electrons are transferred from the TiO2 nanocrystals to the surface via the MtrC conduit. In other words, in these TiO2/MtrC hybrid photodiodes, MtrC traps the conduction-band electrons from TiO2 before transferring them to the electrode, creating a photobioelectrochemical system in which a redox protein is used to mimic the efficient charge separation found in biological photosystems.

5.
Microb Genom ; 6(7)2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32584752

RESUMO

Rearrangements of large genome fragments occur in bacteria between repeat sequences and can impact on growth and gene expression. Homologous recombination resulting in inversion between indirect repeats and excision/translocation between direct repeats enables these structural changes. One form of rearrangement occurs around ribosomal operons, found in multiple copies across many bacteria, but identification of these rearrangements by sequencing requires reads of several thousand bases to span the ribosomal operons. With long-read sequencing aiding the routine generation of complete bacterial assemblies, we have developed socru, a typing method for the order and orientation of genome fragments between ribosomal operons. It allows for a single identifier to convey the order and orientation of genome-level structure and we have successfully applied this typing to 433 of the most common bacterial species. In a focused analysis, we observed the presence of multiple structural genotypes in nine bacterial pathogens, underscoring the importance of routinely assessing this form of variation alongside traditional single-nucleotide polymorphism (SNP) typing.


Assuntos
Bactérias/genética , Óperon , Proteínas Ribossômicas/genética , Análise de Sequência de DNA/métodos , Proteínas de Bactérias/genética , Ordem dos Genes , Proteínas de Choque Térmico , Fragmentos de Peptídeos , Software
6.
Microorganisms ; 8(9)2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32883020

RESUMO

Extensively drug-resistant (XDR) Salmonella Typhi has been reported in Sindh province of Pakistan since 2016. The potential for further spread is of serious concern as remaining treatment options are severely limited. We report the phenotypic and genotypic characterization of 27 XDR S. Typhi isolated from patients attending Jinnah Hospital, Lahore, Pakistan. Isolates were identified by biochemical profiling; antimicrobial susceptibility was determined by a modified Kirby-Bauer method. These findings were confirmed using Illumina whole genome nucleotide sequence data. All sequences were compared to the outbreak strain from Southern Pakistan and typed using the S. Typhi genotyping scheme. All isolates were confirmed by a sequence analysis to harbor an IncY plasmid and the CTX-M-15 ceftriaxone resistance determinant. All isolates were of the same genotypic background as the outbreak strain from Sindh province. We report the first emergence of XDR S. Typhi in Punjab province of Pakistan confirmed by whole genome sequencing.

7.
Sci Rep ; 5: 11677, 2015 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-26126857

RESUMO

Extracellular microbe-mineral electron transfer is a major driving force for the oxidation of organic carbon in many subsurface environments. Extracellular multi-heme cytochromes of the Shewenella genus play a major role in this process but the mechanism of electron exchange at the interface between cytochrome and acceptor is widely debated. The 1.8 Å x-ray crystal structure of the decaheme MtrC revealed a highly conserved CX8C disulfide that, when substituted for AX8A, severely compromised the ability of S. oneidensis to grow under aerobic conditions. Reductive cleavage of the disulfide in the presence of flavin mononucleotide (FMN) resulted in the reversible formation of a stable flavocytochrome. Similar results were also observed with other decaheme cytochromes, OmcA, MtrF and UndA. The data suggest that these decaheme cytochromes can transition between highly reactive flavocytochromes or less reactive cytochromes, and that this transition is controlled by a redox active disulfide that responds to the presence of oxygen.


Assuntos
Proteínas de Bactérias/metabolismo , Flavinas/metabolismo , Minerais/metabolismo , Shewanella/metabolismo , Aerobiose , Motivos de Aminoácidos , Sequência de Aminoácidos , Anaerobiose , Proteínas de Bactérias/química , Cristalografia por Raios X , Citocromos/metabolismo , Dissulfetos/metabolismo , Transporte de Elétrons , Eletroforese em Gel de Poliacrilamida , Glutationa/metabolismo , Heme/metabolismo , Oxirredução , Filogenia , Alinhamento de Sequência , Shewanella/crescimento & desenvolvimento , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta
8.
Arch Dis Child ; 95(8): 639-41, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20371591

RESUMO

Concern exists about measles, mumps and rubella (MMR) vaccine in egg-allergic children, although this has been shown to be safe. Guidelines from the Royal College of Paediatrics and Child Health (RCPCH) and British Society of Allergy and Clinical Immunology (BSACI) suggesting which children should be referred to hospital for MMR, were published in 2000. We audited referrals to hospital for MMR against these guidelines. One hundred and ten children were referred for MMR to Birmingham Heartlands Hospital (2002-2004) and Alder Hey Children's Hospital (2006-2009). Eighty-two (75%) children did not meet the published criteria. Only 13 children (12%) had severe egg allergy. The first dose of MMR vaccine was delayed by >30 days in 81% of children. All children were given MMR, none had a significant reaction. Children with egg allergy do not need to be given MMR in hospital, but MMR is often delayed by unnecessary hospital referral. New BSACI guidelines encouraging MMR vaccination of egg-allergic children in primary care need to be disseminated.


Assuntos
Ovos/efeitos adversos , Hospitalização/estatística & dados numéricos , Vacina contra Sarampo-Caxumba-Rubéola/efeitos adversos , Encaminhamento e Consulta/normas , Pré-Escolar , Inglaterra , Hipersensibilidade Alimentar/complicações , Fidelidade a Diretrizes/normas , Humanos , Lactente , Auditoria Médica , Seleção de Pacientes , Guias de Prática Clínica como Assunto , Encaminhamento e Consulta/estatística & dados numéricos , Procedimentos Desnecessários/estatística & dados numéricos
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