RESUMEN
The next 20 years will see unprecedented growth in Canada's senior population, with higher demands and changing expectations challenging long-term care systems. The Canadian Institute for Health Information (CIHI) linked long-term and acute care data for over 59,000 seniors across six provinces and territories to analyze the pathways and transition patterns of seniors receiving long-term care services. The analysis revealed factors related to residential care entry and identified profiles of seniors admitted into residential care before it may be clinically necessary. This work provides critical information for health system decision-makers to ensure that our long-term care systems are responsive, effective and sustainable.
Asunto(s)
Continuidad de la Atención al Paciente/estadística & datos numéricos , Cuidados a Largo Plazo/estadística & datos numéricos , Anciano , Anciano de 80 o más Años , Canadá/epidemiología , Servicios de Salud para Ancianos , Servicios de Atención de Salud a Domicilio/estadística & datos numéricos , Hogares para Ancianos/estadística & datos numéricos , Hospitalización , HumanosRESUMEN
The formation of an incapacitating biofilm on Caenorhabditis elegans by Yersinia pseudotuberculosis represents a tractable model for investigating the genetic basis for host-pathogen interplay during the biofilm-mediated infection of a living surface. Previously we established a role for quorum sensing (QS) and the master motility regulator, FlhDC, in biofilm formation by Y. pseudotuberculosis on C. elegans. To obtain further genome-wide insights, we used transcriptomic analysis to obtain comparative information on C. elegans in the presence and absence of biofilm and on wild-type Y. pseudotuberculosis and Y. pseudotuberculosis QS mutants. Infection of C. elegans with the wild-type Y. pseudotuberculosis resulted in the differential regulation of numerous genes, including a distinct subset of nematode C-lectin (clec) and fatty acid desaturase (fat) genes. Evaluation of the corresponding C. elegans clec-49 and fat-3 deletion mutants showed delayed biofilm formation and abolished biofilm formation, respectively. Transcriptomic analysis of Y. pseudotuberculosis revealed that genes located in both of the histidine utilization (hut) operons were upregulated in both QS and flhDC mutants. In addition, mutation of the regulatory gene hutC resulted in the loss of biofilm, increased expression of flhDC, and enhanced swimming motility. These data are consistent with the existence of a regulatory cascade in which the Hut pathway links QS and flhDC. This work also indicates that biofilm formation by Y. pseudotuberculosis on C. elegans is an interactive process during which the initial attachment/recognition of Yersinia to/by C. elegans is followed by bacterial growth and biofilm formation.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Caenorhabditis elegans/inmunología , Caenorhabditis elegans/microbiología , Interacciones Huésped-Patógeno , Yersinia pseudotuberculosis/inmunología , Yersinia pseudotuberculosis/fisiología , Animales , Perfilación de la Expresión GénicaRESUMEN
Graves' disease results from thyroid-stimulating Abs (TSAbs) activating the thyrotropin receptor (TSHR). How TSAbs arise from early precursor B cells has not been established. Genetic and environmental factors may contribute to pathogenesis, including the bacterium Yersinia enterocolitica. We developed two pathogenic monoclonal TSAbs from a single experimental mouse undergoing Graves' disease, which shared the same H and L chain germline gene rearrangements and then diversified by numerous somatic hypermutations. To address the Ag specificity of the shared germline precursor of the monoclonal TSAbs, we prepared rFab germline, which showed negligible binding to TSHR, indicating importance of somatic hypermutation in acquiring TSAb activity. Using rFab chimeras, we demonstrate the dominant role of the H chain V region in TSHR recognition. The role of microbial Ags was tested with Y. enterocolitica proteins. The monoclonal TSAbs recognize 37-kDa envelope proteins, also recognized by rFab germline. MALDI-TOF identified the proteins as outer membrane porin (Omp) A and OmpC. Using recombinant OmpA, OmpC, and related OmpF, we demonstrate cross-reactivity of monoclonal TSAbs with the heterogeneous porins. Importantly, rFab germline binds recombinant OmpA, OmpC, and OmpF confirming reactivity with Y. enterocolitica. A human monoclonal TSAb, M22 with similar properties to murine TSAbs, also binds recombinant porins, showing cross-reactivity of a spontaneously arising pathogenic Ab with Y. enterocolitica. The data provide a mechanistic framework for molecular mimicry in Graves' disease, where early precursor B cells are expanded by Y. enterocolitica porins to undergo somatic hypermutation to acquire a cross-reactive pathogenic response to TSHR.
Asunto(s)
Mutación de Línea Germinal , Enfermedad de Graves/etiología , Inmunoglobulinas Estimulantes de la Tiroides/genética , Inmunoglobulinas Estimulantes de la Tiroides/inmunología , Yersinia enterocolitica/inmunología , Anticuerpos Monoclonales/genética , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/inmunología , Proteínas Bacterianas/metabolismo , Expresión Génica , Enfermedad de Graves/genética , Enfermedad de Graves/inmunología , Humanos , Fragmentos Fab de Inmunoglobulinas/genética , Fragmentos Fab de Inmunoglobulinas/inmunología , Fragmentos Fab de Inmunoglobulinas/metabolismo , Inmunoglobulina G/inmunología , Inmunoglobulina G/metabolismo , Región Variable de Inmunoglobulina/genética , Inmunoglobulinas Estimulantes de la Tiroides/metabolismo , Unión Proteica/inmunología , Subunidades de Proteína/inmunología , Subunidades de Proteína/metabolismo , Receptores de Tirotropina/química , Receptores de Tirotropina/inmunología , Receptores de Tirotropina/metabolismo , Proteínas RecombinantesRESUMEN
Ulva zoospores preferentially settle on N-acylhomoserine lactone (AHL) producing marine bacterial biofilms. To investigate whether AHL signal molecules also affect the success and rate of zoospore germination in addition to zoospore attraction, the epiphytic bacteria associated with mature Ulva linza were characterized and bacterial isolates representative of this community tested for the ability to produce AHLs. Two of these AHL-producing isolates, Sulfitobacter spp. 376 and Shewanella spp. 79, were transformed with plasmids expressing the Bacillus spp. AHL lactonase gene aiiA to generate AHL-deficient variants. The germination and growth of U. linza zoospores was studied in the presence of these AHL-deficient strains and their AHL-producing counterparts. This revealed that the AHLs produced by Sulfitobacter spp. and Shewanella spp. or the bacterial products they regulate have a negative impact on both zoospore germination and the early growth of the Ulva germling. Further experiments with Escherichia coli biofilms expressing recombinant AHL synthases and synthetic AHLs provide data to demonstrate that zoospores germinated and grown in the absence of AHLs were significantly longer than those germinated in the presence of AHLs. These results reveal an additional role for AHLsâ per se in the interactive relationships between marine bacteria and Ulva zoospores.
Asunto(s)
Acil-Butirolactonas/química , Biopelículas , Ulva/crecimiento & desarrollo , Ulva/microbiología , Bacillus/enzimología , Escherichia coli/genética , Ligasas/genética , Filogenia , Percepción de Quorum , Rhodobacteraceae/genética , Rhodobacteraceae/crecimiento & desarrollo , Shewanella/genética , Shewanella/crecimiento & desarrolloRESUMEN
The photosynthetic alphaproteobacterium Rhodospirillum rubrum S1H is part of the Micro-Ecological Life Support System Alternative (MELiSSA) project that is aiming to develop a closed life support system for oxygen, water and food production to support human life in space in forthcoming long-term space exploration missions. In the present study, R. rubrum S1H was cultured in a rotating wall vessel (RWV), simulating partial microgravity conditions on Earth. The bacterium showed a significant response to cultivation in simulated microgravity at the transcriptomic, proteomic and metabolic levels. In simulated microgravity conditions three N-acyl-l-homoserine lactones (C10-HSL, C12-HSL and 3-OH-C14-HSL) were detected in concentrations that were twice those detected under normal gravity, while no differences in cell density was detected. In addition, R. rubrum cultivated in modelled microgravity showed higher pigmentation than the normal gravity control, without change in culture oxygenation. When compared to randomized microgravity cultivation using a random positioning machine, significant overlap for the top differentially expressed genes and proteins was observed. Cultivation in this new artificial environment of simulated microgravity showed new properties of this well-known bacterium, including its first, to our knowledge, complete quorum-sensing-related N-acylhomoserine lactone profile.
Asunto(s)
Acil-Butirolactonas/metabolismo , Regulación Bacteriana de la Expresión Génica , Percepción de Quorum , Rhodospirillum rubrum/fisiología , Ingravidez , Perfilación de la Expresión Génica , Metabolómica , Pigmentos Biológicos/metabolismo , Proteoma/análisis , Rhodospirillum rubrum/citologíaRESUMEN
Yersinia pseudotuberculosis forms biofilms on Caenorhabditis elegans which block nematode feeding. This genetically amenable host-pathogen model has important implications for biofilm development on living, motile surfaces. Here we show that Y. pseudotuberculosis biofilm development on C. elegans is governed by N-acylhomoserine lactone (AHL)-mediated quorum sensing (QS) since (i) AHLs are produced in nematode associated biofilms and (ii) Y. pseudotuberculosis strains expressing an AHL-degrading enzyme or in which the AHL synthase (ypsI and ytbI) or response regulator (ypsR and ytbR) genes have been mutated, are attenuated. Although biofilm formation is also attenuated in Y. pseudotuberculosis strains carrying mutations in the QS-controlled motility regulator genes, flhDC and fliA, and the flagellin export gene, flhA, flagella are not required since fliC mutants form normal biofilms. However, in contrast to the parent and fliC mutant, Yop virulon proteins are up-regulated in flhDC, fliA and flhA mutants in a temperature and calcium independent manner. Similar observations were found for the Y. pseudotuberculosis QS mutants, indicating that the Yop virulon is repressed by QS via the master motility regulator, flhDC. By curing the pYV virulence plasmid from the ypsI/ytbI mutant, by growing YpIII under conditions permissive for type III needle formation but not Yop secretion and by mutating the type III secretion apparatus gene, yscJ, we show that biofilm formation can be restored in flhDC and ypsI/ytbI mutants. These data demonstrate that type III secretion blocks biofilm formation and is reciprocally regulated with motility via QS.
Asunto(s)
Biopelículas/crecimiento & desarrollo , Caenorhabditis elegans/fisiología , Regulación Bacteriana de la Expresión Génica/fisiología , Interacciones Huésped-Patógeno/genética , Percepción de Quorum/genética , Yersinia pseudotuberculosis/genética , Acil-Butirolactonas/metabolismo , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Caenorhabditis elegans/microbiología , Proteínas Motoras Moleculares/genética , Proteínas Motoras Moleculares/metabolismo , MutaciónRESUMEN
The distribution and transmission of Yersinia pestis, the bacterial agent of plague, responds dynamically to climate, both within wildlife reservoirs and human populations. The exact mechanisms mediating plague's response to climate are still poorly understood, particularly across large environmentally heterogeneous regions encompassing several reservoir species. A heterogeneous response to precipitation was observed in plague intensity across northern and southern China during the Third Pandemic. This has been attributed to the response of reservoir species in each region. We use environmental niche modelling and hindcasting methods to test the response of a broad range of reservoir species to precipitation. We find little support for the hypothesis that the response of reservoir species to precipitation mediated the impact of precipitation on plague intensity. We instead observed that precipitation variables were of limited importance in defining species niches and rarely showed the expected response to precipitation across northern and southern China. These findings do not suggest that precipitation-reservoir species dynamics never influence plague intensity but that instead, the response of reservoir species to precipitation across a single biome cannot be assumed and that limited numbers of reservoir species may have a disproportional impact upon plague intensity.
RESUMEN
Since its first identification in 1894 during the third pandemic in Hong Kong, there has been significant progress of understanding the lifestyle of Yersinia pestis, the pathogen that is responsible for plague. Although we now have some understanding of the pathogen's physiology, genetics, genomics, evolution, gene regulation, pathogenesis and immunity, there are many unknown aspects of the pathogen and its disease development. Here, we focus on some of the knowns and unknowns relating to Y. pestis and plague. We notably focus on some key Y. pestis physiological and virulence traits that are important for its mammal-flea-mammal life cycle but also its emergence from the enteropathogen Yersinia pseudotuberculosis. Some aspects of the genetic diversity of Y. pestis, the distribution and ecology of plague as well as the medical countermeasures to protect our population are also provided. Lastly, we present some biosafety and biosecurity information related to Y. pestis and plague.
RESUMEN
BACKGROUND: Cell-to-cell communication (quorum sensing (QS)) co-ordinates bacterial behaviour at a population level. Consequently the behaviour of a natural multi-species community is likely to depend at least in part on co-existing QS and quorum quenching (QQ) activities. Here we sought to discover novel N-acylhomoserine lactone (AHL)-dependent QS and QQ strains by investigating a bacterial community associated with the rhizosphere of ginger (Zingiber officinale) growing in the Malaysian rainforest. RESULTS: By using a basal growth medium containing N-(3-oxohexanoyl)homoserine lactone (3-oxo-C6-HSL) as the sole source of carbon and nitrogen, the ginger rhizosphere associated bacteria were enriched for strains with AHL-degrading capabilities. Three isolates belonging to the genera Acinetobacter (GG2), Burkholderia (GG4) and Klebsiella (Se14) were identified and selected for further study. Strains GG2 and Se14 exhibited the broadest spectrum of AHL-degrading activities via lactonolysis while GG4 reduced 3-oxo-AHLs to the corresponding 3-hydroxy compounds. In GG2 and GG4, QQ was found to co-exist with AHL-dependent QS and GG2 was shown to inactivate both self-generated and exogenously supplied AHLs. GG2, GG4 and Se14 were each able to attenuate virulence factor production in both human and plant pathogens. CONCLUSIONS: Collectively our data show that ginger rhizosphere bacteria which make and degrade a wide range of AHLs are likely to play a collective role in determining the QS-dependent phenotype of a polymicrobial community.
Asunto(s)
Acinetobacter/crecimiento & desarrollo , Acil-Butirolactonas/metabolismo , Burkholderia/crecimiento & desarrollo , Percepción de Quorum , Rizosfera , Zingiber officinale/microbiología , Acinetobacter/aislamiento & purificación , Acinetobacter/metabolismo , Burkholderia/aislamiento & purificación , Burkholderia/metabolismo , Medios de Cultivo , MalasiaRESUMEN
BACKGROUND: In order to identify new virulence determinants in Y. pseudotuberculosis a comparison between its genome and that of Yersinia pestis was undertaken. This reveals dozens of pseudogenes in Y. pestis, which are still putatively functional in Y. pseudotuberculosis and may be important in the enteric lifestyle. One such gene, YPTB1572 in the Y. pseudotuberculosis IP32953 genome sequence, encodes a protein with similarity to invasin, a classic adhesion/invasion protein, and to intimin, the attaching and effacing protein from enteropathogenic (EPEC) and enterohaemorraghic (EHEC) Escherichia coli. RESULTS: We termed YPTB1572 Ifp (Intimin family protein) and show that it is able to bind directly to human HEp-2 epithelial cells. Cysteine and tryptophan residues in the C-terminal region of intimin that are essential for function in EPEC and EHEC are conserved in Ifp. Protein binding occurred at distinct foci on the HEp-2 cell surface and can be disrupted by mutation of a single cysteine residue at the C-terminus of the protein. Temporal expression analysis using lux reporter constructs revealed that ifp is expressed at late log phase at 37°C in contrast to invasin, suggesting that Ifp is a late stage adhesin. An ifp defined mutant showed a reduction in adhesion to HEp-2 cells and was attenuated in the Galleria mellonella infection model. CONCLUSION: A new Y. pseudotuberculosis adhesin has been identified and characterised. This Ifp is a new member in the family of invasin/intimin outer membrane adhesins.
Asunto(s)
Adhesinas Bacterianas/genética , Adhesinas Bacterianas/metabolismo , Factores de Virulencia/genética , Factores de Virulencia/metabolismo , Yersinia pseudotuberculosis/genética , Yersinia pseudotuberculosis/patogenicidad , Sustitución de Aminoácidos/genética , Animales , Adhesión Bacteriana , Línea Celular , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Hepatocitos/microbiología , Humanos , Lepidópteros/microbiología , Mutagénesis Sitio-Dirigida , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Unión Proteica , Análisis de SupervivenciaRESUMEN
Transcription of the 74 kb Pseudomonas fluorescens mupirocin [pseudomonic acid (PA)] biosynthesis cluster depends on quorum sensing-dependent regulation via the LuxI/LuxR homologues MupI/MupR. To facilitate analysis of novel PAs from pathway mutants, we investigated factors that affect mup gene expression. First, the signal produced by MupI was identified as N-(3-oxodecanoyl)homoserine lactone, but exogenous addition of this molecule did not activate mupirocin production prematurely nor did expression of mupI in trans increase metabolite production. Second, we confirmed that mupX, encoding an amidase/hydrolase that can degrade N-acylhomoserine lactones, is also required for efficient expression, consistent with its occurrence in a regulatory module linked to unrelated genes in P. fluorescens. Third, and most significantly, mupR expression in trans to wild type and mutants can increase production of antibiotic and novel intermediates up to 17-fold.
Asunto(s)
Regulación Bacteriana de la Expresión Génica , Mupirocina/metabolismo , Pseudomonas fluorescens/fisiología , Percepción de Quorum , Regulación hacia Arriba , Antibacterianos/biosíntesis , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Pseudomonas fluorescens/genéticaRESUMEN
Quorum sensing (QS) in Yersinia pseudotuberculosis involves two pairs of LuxRI orthologues (YpsRI and YtbRI) and multiple N-acylhomoserine lactones (AHLs). In a ypsI/ytbI mutant, AHL synthesis was abolished, unaffected in a ypsR/ytbR double mutant and substantially reduced in a ypsI/ytbR mutant, indicating that neither YpsR nor YtbR is essential for AHL synthesis. To determine the interrelationship between YpsRI and YtbRI we constructed chromosomal lux-promoter fusions to ypsR, ypsI, ytbR and ytbI and examined their expression in each of the QS mutant backgrounds. The YpsRI system negatively autoregulates itself but positively regulates the expression of the ytbRI system whereas the ytbRI system is positively autoregulated but only at the level of ytbI expression. YtbRI does not control expression of ypsR or ypsI. This hierarchical QS system controls swimming motility via regulation of flhDC and fliA. The AHLs synthesized via YtbI play a dual role, activating flhDC, in conjunction with YpsR but repressing fliA in conjunction with YtbR and YpsR. In liquid and plate assays, the early onset of motility observed in ypsR and ypsI mutants was abolished in ytbI, ytbR ypsI/ytbI, ypsR/ytbR mutants, indicating that QS regulates motility both positively (via YtbRI) and negatively (via YpsRI).
Asunto(s)
Proteínas Bacterianas/metabolismo , Flagelos/metabolismo , Regulación Bacteriana de la Expresión Génica , Percepción de Quorum , Factor sigma/metabolismo , Yersinia pseudotuberculosis/fisiología , Acil-Butirolactonas/metabolismo , Proteínas Bacterianas/genética , Secuencia de Consenso , Medios de Cultivo/análisis , Flagelos/genética , Operón , Regiones Promotoras Genéticas , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Factor sigma/genética , Transactivadores/genética , Transactivadores/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Yersinia pseudotuberculosis/genéticaRESUMEN
N-acylhomoserine lactone (AHL) quorum-sensing molecules modulate the swimming behaviour of zoospores of the macroalga Ulva to facilitate the location of bacterial biofilms. Here we show that the intertidal surfaces colonized by Ulva are dominated by Alphaproteobacteria, particularly the Rhodobacteraceae family, and the Bacteroidetes family Flavobacteriaceae, and that this diverse assemblage both produces and degrades AHLs. N-acylhomoserine lactones could also be extracted from the surfaces of pebbles recovered from intertidal rock-pools. Bacteria representative of this assemblage were isolated and tested for the production and degradation of AHLs, and for their ability to modulate zoospore settlement at different biofilm densities. Of particular interest was a Shewanella sp. This strain produced three major AHLs (OC4, OC10 and OC12) in the late exponential phase, but the longer-chain AHLs were rapidly degraded in the stationary phase. Degradation occurred via both lactonase and amidase activity. A close relationship was found between AHL synthesis and Ulva zoospore settlement. The Shewanella isolate also interfered with AHL production by a Sulfitobacter isolate and its ability to enhance zoospore settlement in a polymicrobial biofilm. This influence on the attachment of Ulva zoospores suggests that AHL-degrading strains can affect bacterial community behaviour by interfering with quorum sensing between neighbouring bacteria. More importantly, these interactions may exert wider ecological effects across different kingdoms.
Asunto(s)
Acil-Butirolactonas/metabolismo , Microbiología Ambiental , Percepción de Quorum , Rhodobacteraceae/fisiología , Shewanella/fisiología , Ulva/fisiología , Antibiosis , Biodiversidad , Biopelículas/crecimiento & desarrollo , Ecosistema , Rhodobacteraceae/enzimología , Rhodobacteraceae/crecimiento & desarrollo , Rhodobacteraceae/metabolismo , Shewanella/enzimología , Shewanella/crecimiento & desarrollo , Shewanella/metabolismo , Ulva/crecimiento & desarrollo , Ulva/metabolismoRESUMEN
OmpR has been demonstrated to negatively regulate the expression of the flagellar master operon flhDC in a wide variety of bacterial species. Here we report the positive regulation of flhDC expression by OmpR in Yersinia pseudotuberculosis. A sigma(70)-dependent promoter was identified by primer extension analysis and an active region with two conserved OmpR-binding sites around the flhDC promoter was confirmed. To confirm the regulation of flhDC expression by OmpR, flhDC as well as the downstream flagellar genes fliA, flgD, flgA, flgM, fliC and flaA were fused to lacZ, and decreased expression of all these genes in an ompR mutant (Delta ompR) was detected. Furthermore, Delta ompR was defective in bacterial motility and flagella synthesis. This defect was due to the low level of expression of flhDC in Delta ompR since overproduction of FlhDC in Delta ompR restored bacterial motility. The importance of two conserved OmpR-binding sites around the flhDC promoter region in the regulation of flhDC expression by OmpR was demonstrated by the fact that mutation of either one or both sites significantly decreased the promoter activity in the wild-type but not in Delta ompR. The binding of OmpR to these two sites was also demonstrated by DNA mobility shift assay. The possible mechanism underlying this positive regulation in Y. pseudotuberculosis is discussed. To our knowledge, this is the first report to demonstrate that OmpR positively regulates flhDC expression.
Asunto(s)
Proteínas Bacterianas/metabolismo , Operón , Transactivadores/metabolismo , Yersinia pseudotuberculosis/genética , Proteínas Bacterianas/genética , Flagelos/genética , Flagelos/metabolismo , Regulación Bacteriana de la Expresión Génica , Mutación , Regiones Promotoras Genéticas , Transactivadores/genética , Sitio de Iniciación de la Transcripción , Yersinia pseudotuberculosis/metabolismoRESUMEN
Although Yersinia enterocolitica genomes are highly heterogeneous, they contain a conserved N-acylhomoserine lactone-dependent (AHL) quorum sensing (QS) system consisting of the luxR and luxI orthologs yenR and yenI respectively. Certain hypervirulent strains also contain a putative orphan luxR gene, ycoR, that is not linked to an AHL synthase. To explore the contribution of yenR/yenI/ycoR to QS-dependent phenotypes in Yersinia enterocolitica strain 8081, single and multiple mutants were constructed. AHL profiling identified N-(3-oxohexanoyl) homoserine lactone, N-hexanoylhomoserine lactone, and N-(3-oxoseptanoyl) homoserine lactone as the most abundant. The AHL profiles of the yenR, ycoR and yenR/ycoR mutants were similar to the parent suggesting that the two LuxR homologues do not regulate AHL production while the yenI mutants were AHL-negative. A role for QS in swimming motility and cell attachment was demonstrated. Down-regulation of the virulence plasmid partition gene, spyA, in yenI and yenI/yenR/ycoR mutants is consistent with the greater loss of the Y. enterocolitica pYVe virulence plasmid in the yenI mutant during serial passage at 37 °C but not at 22 °C. A role for QS-regulated spyA in virulence plasmid maintenance is suggested.
RESUMEN
The human pathogens Yersinia pseudotuberculosis and Yersinia enterocolitica cause enterocolitis, while Yersinia pestis is responsible for pneumonic, bubonic, and septicaemic plague. All three share an infection strategy that relies on a virulence factor arsenal to enable them to enter, adhere to, and colonise the host while evading host defences to avoid untimely clearance. Their arsenal includes a number of adhesins that allow the invading pathogens to establish a foothold in the host and to adhere to specific tissues later during infection. When the host innate immune system has been activated, all three pathogens produce a structure analogous to a hypodermic needle. In conjunction with the translocon, which forms a pore in the host membrane, the channel that is formed enables the transfer of six 'effector' proteins into the host cell cytoplasm. These proteins mimic host cell proteins but are more efficient than their native counterparts at modifying the host cell cytoskeleton, triggering the host cell suicide response. Such a sophisticated arsenal ensures that yersiniae maintain the upper hand despite the best efforts of the host to counteract the infecting pathogen.
RESUMEN
Many bacterial species communicate using a complex system known as quorum sensing (QS) in which gene expression is controlled in response to cell density. In this study an N-acylhomoserine lactone (AHL) synthase (Rru_A3396) knockout mutant (M68) of Rhodospirillum rubrum S1H (WT) was constructed and characterized phenotypically under light anaerobic conditions. Results showed that R. rubrum WT produces unsubstituted, 3-OH and 3-oxo-substituted AHLs with acyl chains ranging from 4 to 14 carbons, with 3-OH-C8 being the most abundant. Growth, pigment content and swimming motility were found to be under the control of this LuxI-type QS system. In addition, cultivation in a low shear environment put forward the aggregative phenotype of M68 and linked biofilm formation to QS in R. rubrum S1H. Interestingly, QS-mutant M68 continued to produce decreased levels of 3-OH-C8-HSL, probably due to the presence of an extra HdtS-type AHL synthase.
Asunto(s)
Técnicas de Inactivación de Genes , Ligasas/metabolismo , Percepción de Quorum , Rhodospirillum rubrum/fisiología , Acil-Butirolactonas/química , Acil-Butirolactonas/metabolismo , Anaerobiosis , Ligasas/genética , Locomoción , Pigmentos Biológicos/metabolismo , Rhodospirillum rubrum/genética , Rhodospirillum rubrum/metabolismoRESUMEN
A new class of bacteria-attachment-resistant materials is discovered using a multi-generation polymer microarray methodology that reduces bacterial attachment by up to 99.3% compared with a leading commercially available silver hydrogel anti-bacterial material. The coverage of three bacterial species, Pseudomonas aeruginosa, Staphylococcus aureus, and uropathogenic Escherichia coli is assessed.
Asunto(s)
Adhesión Bacteriana/efectos de los fármacos , Diseño de Fármacos , Análisis por Micromatrices , Ácidos Polimetacrílicos/farmacología , Ácidos Polimetacrílicos/química , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/fisiología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/fisiologíaRESUMEN
The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a combinatorial library of materials to be screened in a parallel, high throughput format (1). Herein is described the formation and characterization of a polymer microarray using an on-chip photopolymerization technique (2). This involves mixing monomers at varied ratios to produce a library of monomer solutions, transferring the solution to a glass slide format using a robotic printing device and curing with UV irradiation. This format is readily amenable to many biological assays, including stem cell attachment and proliferation, cell sorting and low bacterial adhesion, allowing the ready identification of 'hit' materials that fulfill a specific biological criterion (3-5). Furthermore, the use of high throughput surface characterization (HTSC) allows the biological performance to be correlated with physio-chemical properties, hence elucidating the biological-material interaction (6). HTSC makes use of water contact angle (WCA) measurements, atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). In particular, ToF-SIMS provides a chemically rich analysis of the sample that can be used to correlate the cell response with a molecular moiety. In some cases, the biological performance can be predicted from the ToF-SIMS spectra, demonstrating the chemical dependence of a biological-material interaction, and informing the development of hit materials (5,3).