Bacterial adherence and hemolysin production from Escherichia coli induces histamine and leukotriene release from various cells

We investigated the role of bacterial adherence and hemolysin production from Escherichia coli parent and genetically cloned strains as to their eft'ects on bistaJidne release from rat mast cells and leukotriene generation from human polymorphonuclear granulocytes. These mediators were involved in the induction of inftammatory disease processes and led, for example, to enhancement of vascular permeability, chemotaxis (leukotriene 84 [LTB4]), chemoaggregation, lysosomal enzyme release, and smooth muscle contraction, (LTC4, LTD4 , and LTE4). Washed bacteria (E. coli K-12 Ms+ my=; E. coli 536 Ms+ MR= my=) as weil as their culture supematants were analyzed. Washed E. coli K-12 (Hiy+), unlike Hly- strains, induced high amounts of histamine release from rat mast cells and chemotactic activity from human polymorphonuclear granulocytes. Significant leukotriene releasewas obtained with washed E. coli K-12 my+ strains and their bacterial culture supematants. Leukotriene induction was dependent on the amount of hemolysin activity present in the supematant. However, additional soluble factors should also be considered. The presence of hemolysin appeared to aceeierate and enhance the rate of phagocytosis of bacteria by neutrophUs. When E. coli 536 (MS+ MR= Hly=) strains were analyzed, the simultaneous presence of MR+ pili and hemolysin production led to an increase in histamine release as compared with MR- my+ strains. The genetically cloned MR+ my+ E. coli 536 strain induced higher amounts of IeukotrieDes as compared with the wUd-type strain. Our data soggest a potent role for adhesins and hemolysin as virulence factors in inducing the release of inftammatory mediators

Distribution, expression and long range mapping of legiolysin gene (lly) specific DNA sequences in Legionellae

The legiolysin gene (lly) cloned from Legionella pneumophila Philadelphia 1 confers the phenotypes of hemolysis and browning of the culture medium. An internal Uy-specific DNA probe was used in Southern hybridizations for the detection of Uy-specific DNA in the genomes of legioneUae and other gram-negative pathogenic bacteria. Under conditi9ns of high stringency, tlie Uy DNA probe specifically reacted with DNA fragments fr9m L. pneumophiüz isolates; by reducing stringency, hybridization was also observed for all other Legionella strains tested. No hybridization occurred with DNAs isolated from bact~ria of other genera. The Uy genewas mapped by pulsed-field gel electrophoresis to the respective genomic Notl fragments of Legionelltz isolates. By using antilegiolysin monospecific polyclonal antibodies in Western blots (immunoblots), Lly proteins could be detected only in L. pneumophila isolates

Toward vicarious calibration of microwave remote-sensing satellites in arid environments

The Soil Moisture and Ocean Salinity (SMOS) satellite marks the commencement of dedicated global surface soil moisture missions, and the first mission to make passive microwave observations at L-band. On-orbit calibration is an essential part of the instrument calibration strategy, but on-board beam-filling targets are not practical for such large apertures. Therefore, areas to serve as vicarious calibration targets need to be identified. Such sites can only be identified through field experiments including both in situ and airborne measurements. For this purpose, two field experiments were performed in central Australia. Three areas are studied as follows: 1) Lake Eyre, a typically dry salt lake; 2) Wirrangula Hill, with sparse vegetation and a dense cover of surface rock; and 3) Simpson Desert, characterized by dry sand dunes. Of those sites, only Wirrangula Hill and the Simpson Desert are found to be potentially suitable targets, as they have a spatial variation in brightness temperatures of <4 K under normal conditions. However, some limitations are observed for the Simpson Desert, where a bias of 15 K in vertical and 20 K in horizontal polarization exists between model predictions and observations, suggesting a lack of understanding of the underlying physics in this environment. Subsequent comparison with model predictions indicates a SMOS bias of 5 K in vertical and 11 K in horizontal polarization, and an unbiased root mean square difference of 10 K in both polarizations for Wirrangula Hill. Most importantly, the SMOS observations show that the brightness temperature evolution is dominated by regular seasonal patterns and that precipitation events have only little impact

Mobilisation and remobilisation of a large archetypal pathogenicity island of uropathogenic Escherichia coli in vitro support the role of conjugation for horizontal transfer of genomic islands

Background: A substantial amount of data has been accumulated supporting the important role of genomic islands (GEIs) - including pathogenicity islands (PAIs) - in bacterial genome plasticity and the evolution of bacterial pathogens. Their instability and the high level sequence similarity of different (partial) islands suggest an exchange of PAIs between strains of the same or even different bacterial species by horizontal gene transfer (HGT). Transfer events of archetypal large genomic islands of enterobacteria which often lack genes required for mobilisation or transfer have been rarely investigated so far. Results: To study mobilisation of such large genomic regions in prototypic uropathogenic E. coli (UPEC) strain 536, PAI II536 was supplemented with the mobRP4 region, an origin of replication (oriVR6K), an origin of transfer (oriTRP4) and a chloramphenicol resistance selection marker. In the presence of helper plasmid RP4, conjugative transfer of the 107-kb PAI II536 construct occured from strain 536 into an E. coli K-12 recipient. In transconjugants, PAI II536 existed either as a cytoplasmic circular intermediate (CI) or integrated site-specifically into the recipient’s chromosome at the leuX tRNA gene. This locus is the chromosomal integration site of PAI II536 in UPEC strain 536. From the E. coli K-12 recipient, the chromosomal PAI II536 construct as well as the CIs could be successfully remobilised and inserted into leuX in a PAI II536 deletion mutant of E. coli 536. Conclusions: Our results corroborate that mobilisation and conjugal transfer may contribute to evolution of bacterial pathogens through horizontal transfer of large chromosomal regions such as PAIs. Stabilisation of these mobile genetic elements in the bacterial chromosome result from selective loss of mobilisation and transfer functions of genomic islands

A strategy for assessing continuity in terrestrial and maritime landscapes from Murujuga (Dampier Archipelago), North West Shelf, Australia

Over the last 20,000 years one-third of the continental land mass of Australia, or 2.12 million km2, has been drowned by postglacial sea-level rise. Much of this drowned territory is thought to have been occupied by humans and where archaeological remains have survived inundation, they can be investigated by underwater and airborne remote sensing, survey and ground-truthing. This study of the Dampier Archipelago of NW Australia is contextualised by a review of the current state of the art of underwater prehistory. In the absence of known sites, we propose terrestrial analogy as a predictive tool for targeting submerged archaeological sites. Geological and topographic contexts are important for assessing preservation potential as is identifying landforms and features around which people may have focused their occupation. Analysis of over 2,500 known archaeological sites from the extraordinarily rich Dampier Archipelago reveals that the vast majority are rock art sites but these are interspersed by a significant number of artefact scatters, myriad stone structures, shell middens and quarry and reduction areas. The majority of these sites are focused on coastal and interior valleys, associated uplands and coastal embayments. While over two-thirds of sites occur on granophyre and basalt substrates, the others are located on quaternary sediments. Regional research on nearby continental islands shows that use of these environments can be expected to pre-date sea level rise. The most likely submerged sites include (i) compacted middens associated with rock pools and estuarine features; (ii) stone structures with associated midden on limestone pavements or with granophyre and basalt boulder fields; (iii) buried midden and other occupation deposits on protected sand sheets; (iv) quarry outcrops, extraction pits and associated reduction debris in areas of fine-grained granophyre and basalt; and (v) midden in consolidated calcarenite shoreline contexts to the north and west of the volcanic suites of the Dampier Archipelago

Uropathogenic E. coli Induce Different Immune Response in Testicular and Peritoneal Macrophages: Implications for Testicular Immune Privilege

Infertility affects one in seven couples and ascending bacterial infections of the male genitourinary tract by Escherichia coli are an important cause of male factor infertility. Thus understanding mechanisms by which immunocompetent cells such as testicular macrophages (TM) respond to infection and how bacterial pathogens manipulate defense pathways is of importance. Whole genome expression profiling of TM and peritoneal macrophages (PM) infected with uropathogenic E. coli (UPEC) revealed major differences in regulated genes. However, a multitude of genes implicated in calcium signaling pathways was a common feature which indicated a role of calcium-dependent nuclear factor of activated T cells (NFAT) signaling. UPEC-dependent NFAT activation was confirmed in both cultured TM and in TM in an in vivo UPEC infectious rat orchitis model. Elevated expression of NFATC2-regulated anti-inflammatory cytokines was found in TM (IL-4, IL-13) and PM (IL-3, IL-4, IL-13). NFATC2 is activated by rapid influx of calcium, an activity delineated to the pore forming toxin alpha-hemolysin by bacterial mutant analysis. Alpha-hemolysin suppressed IL-6 and TNF-α cytokine release from PM and caused differential activation of MAP kinase and AP-1 signaling pathways in TM and PM leading to reciprocal expression of key pro-inflammatory cytokines in PM (IL-1α, IL-1β, IL-6 downregulated) and TM (IL-1β, IL-6 upregulated). In addition, unlike PM, LPS-treated TM were refractory to NFκB activation shown by the absence of degradation of IκBα and lack of pro-inflammatory cytokine secretion (IL-6, TNF-α). Taken together, these results suggest a mechanism to the conundrum by which TM initiate immune responses to bacteria, while maintaining testicular immune privilege with its ability to tolerate neo-autoantigens expressed on developing spermatogenic cells

An Antimicrobial Peptide Regulates Tumor-Associated Macrophage Trafficking via the Chemokine Receptor CCR2, a Model for Tumorigenesis

Tumor-associated macrophages (TAMs) constitute a significant part of infiltrating inflammatory cells that are frequently correlated with progression and poor prognosis of a variety of cancers. Tumor cell-produced human β-defensin-3 (hBD-3) has been associated with TAM trafficking in oral cancer; however, its involvement in tumor-related inflammatory processes remains largely unknown., applying a cross-desensitization strategy of CCR2 and its pharmacological inhibitor (RS102895), respectively, was also carried out. outcome and demonstrates the importance of the innate immune system in the development of tumors

Activity of Desacetylcefotaxime, a Metabolite of Cefotaxime, In Vitro and in a Model of Experimental Infection with Escherichia coli

Die MHK-Werte von Desacetylcefotaxim gegen verschiedene, z. T. ampicillinresistente Stämme von Escherichia coH, die mit Hilfe einer Agardilutionsmethode erhoben wurden, waren höher als die von Cefotaxim und Ceftriaxon, jedoch niedriger als die von Cefoxitin. In einem Modell der systemischen Infektion der Maus mit einem plasmidtragenden, betalactamaseproduzierenden Stamm von E. coli führte die Therapie mit Desacetylcefotaxim zu einer starken Reduktion der Keime pro Leber. Im Vergleich zur Therapie mit Cefotaxim trat die protektive Wirkung aber verlangsamt ein. Desacetylcefotaxim ist also kein unnützes Abbauprodukt von Cefotaxim.The MIC values of desacetylcefotaxime, as determined by an agar dilution method, for several strains of Escherichia coli largely resistant to ampicillin were considerably higher than those of cefotaxime or ceftriaxone respectively. The values were, however, definitely lower than those of cefoxitin. In a mouse model of a systemic infection with a plasmid-bearing, j3-lactamase producing strain of E. coli the therapeutic potency of ampicillin, cefotaxime and desacetylcefotaxime was determined by counting the bacterial numbers per liver at days 1,3 and 7 after subcutaneous infection with about 103 viable bacteria. Desacety1cefotaxime was able to redure thc bacterial load considerably, though with a delay in comparison to cefotaxime