Role of Bacterial Surface Structures on the Interaction of Klebsiella pneumoniae with Phagocytes

Phagocytosis is a key process of the immune system. The human pathogen Klebsiella pneumoniae is a well known example of a pathogen highly resistant to phagocytosis. A wealth of evidence demonstrates that the capsule polysaccharide (CPS) plays a crucial role in resistance to phagocytosis. The amoeba Dictyostelium discoideum shares with mammalian macrophages the ability to phagocytose and kill bacteria. The fact that K. pneumoniae is ubiquitous in nature and, therefore, should avoid predation by amoebae, poses the question whether K. pneumoniae employs similar means to counteract amoebae and mammalian phagocytes. Here we developed an assay to evaluate K. pneumoniae-D. discoideum interaction. The richness of the growth medium affected the threshold at which the cps mutant was permissive for Dictyostelium and only at lower nutrient concentrations the cps mutant was susceptible to predation by amoebae. Given the critical role of bacterial surface elements on host-pathogen interactions, we explored the possible contribution of the lipopolysaccharide (LPS) and outer membrane proteins (OMPs) to combat phagoyctosis by D. discoideum. We uncover that, in addition to the CPS, the LPS O-polysaccharide and the first core sugar participate in Klebsiella resistance to predation by D. discoideum. K. pneumoniae LPS lipid A decorations are also necessary to avoid predation by amoebae although PagP-dependent palmitoylation plays a more important role than the lipid A modification with aminoarabinose. Mutants lacking OMPs OmpA or OmpK36 were also permissive for D. discoideium growth. Except the LPS O-polysaccharide mutants, all mutants were more susceptible to phagocytosis by mouse alveolar macrophages. Finally, we found a correlation between virulence, using the pneumonia mouse model, and resistance to phagocytosis. Altogether, this work reveals novel K. pneumoniae determinants involved in resistance to phagocytosis and supports the notion that Dictyostelium amoebae might be useful as host model to measure K. pneumoniae virulence and not only phagocytosis. © 2013 March et al.

Modeling Klebsiella pneumoniae pathogenesis by infection of the wax moth Galleria mellonella

The implementation of infection models that approximate human disease is essential for understanding pathogenesis at the molecular level and for testing new therapies before they are entered into clinical stages. Insects are increasingly being used as surrogate hosts because they share, with mammals, essential aspects of the innate immune response to infections. We examined whether the larva of the wax moth Galleria mellonella could be used as a host model to conceptually approximate Klebsiella pneumoniae-triggered pneumonia. We report that the G. mellonella model is capable of distinguishing between pathogenic and nonpathogenic Klebsiella strains. Moreover, K. pneumoniae infection of G. mellonella models some of the known features of Klebsiella-induced pneumonia, i.e., cell death associated with bacterial replication, avoidance of phagocytosis by phagocytes, and the attenuation of host defense responses, chiefly the production of antimicrobial factors. Similar to the case for the mouse pneumonia model, activation of innate responses improved G. mellonella survival against subsequent Klebsiella challenge. Virulence factors necessary in the mouse pneumonia model were also implicated in the Galleria model. We found that mutants lacking capsule polysaccharide, lipid A decorations, or the outer membrane proteins OmpA and OmpK36 were attenuated in Galleria. All mutants activated G. mellonella defensive responses. The Galleria model also allowed us to monitor Klebsiella gene expression. The expression levels of cps and the loci implicated in lipid A remodeling peaked during the first hours postinfection, in a PhoPQ- and PmrAB-governed process. Taken together, these results support the utility of G. mellonella as a surrogate host for assessing infections with K. pneumoniae.

The PRIME 1996 cruise: an overview

The UK PRIME cruise, June-July 1996 in the NE Atlantic, consisted of two legs. During the first, detailed chemical and biological observations were made in time-series mode adjacent to the centre of a cold-core eddy in the vicinity of 59 degreesN 20 degreesW using SF6 tracer techniques as the basis for the Lagrangian study. The eddy, which appeared to have been formed the previous winter, remained coherent over the 9 days of the survey and advected only slowly. The phytoplankton community in the eddy was dominated by the coccolithophorid Coccolithus huxleyi. High microzooplankton grazing rates indicated minimal export losses from the surface layer. Significant shifts in many, but not all, of the chemical and biological properties measured were observed over the course of the experiment, especially after the passage of a storm event, which resulted in considerable deepening of the mixed layer followed by a return to fully stratified conditions. The second leg consisted of a transect from 59 degreesN 20 degreesW to 37 degreesN 19 degreesW, with a further Lagrangian time-series study based on a drogue marker initiated at the southern end of the transect. Maximal biological activity was generally encountered in the region between two fronts located at 52.5 degreesN and 48 degreesN, while to the south of 48 degreesN oligotrophic conditions prevailed. At the southern Lagrangian site, a deep chlorophyll maximum was present and high column new production was recorded as a result of the euphotic zone extending below the depth of the nutricline. Microzooplankton grazing rates were lower at this location than at the northern eddy site. The influx of a warm, saline water body into the upper layers during the southern survey led to a major shift in many of the biological and chemical properties being measured. At both the northern and southern Lagrangian sites, the biomass of the mesozooplankton exceeded that of the microzooplankton. (C) 2001 Elsevier Science Ltd. All rights reserved.

Comparisons of plankton at 59 degrees N and 37 degrees N during two Lagrangian drift experiments in the North Atlantic in June and July 1996

Data are summarised for two Lagrangian experiments in the North Atlantic in early summer 1996. At 59 degreesN 20 degreesW, plankton dynamics was studied in an SF, tracer release experiment within a mesoscale eddy over a 9-day period. At 37 degreesN 20 degreesW, a second experiment followed a drifting buoy for 7 days. The data obtained in these two experiments have been averaged for 3 depth strata; the euphotic zone, the surface mixed layer (SML), and the seasonal thermocline immediately beneath the surface mixed layer. At 59 degreesN, the euphotic zone was only marginally deeper than the SML, but at 37 degreesN the SML was ca 30 m and the euphotic depth was ca 110 m. At 37 degreesN, nutrient concentrations in the SML were low but significant new production occurred in the thermocline because of light penetration into the nutricline. The particulate organic carbon (POC) concentration of the SML at 59 degreesN was 13-15 mu mol C kg(-1), but at 37 degreesN POC concentrations were 4 mu mol C kg(-1). These POC measurements include biota and detritus. As a way of investigating latitudinal differences in the plankton communities, estimates have been made of the carbon and nitrogen content of phytoplankton, bacterioplankton, microzooplankton and mesozooplankton. At both 59 degreesN and 37 degreesN, phytoplankton was the largest component, accounting for ca 50% of the planktonic biomass in the SML. At 59 degreesN, microzooplankton was 16% of the planktonic carbon, but at 37 degreesN this reduced to 8% of the total. Mesozooplankton was a relatively constant proportion (ca 20%) of the planktonic carbon in the SML at both 59 degreesN and 37 degreesN. Bacterioplankton was 14% of the biomass at 59 degreesN, increasing to 24% in the microbial loop-dominated system at 37 degreesN. Mean carbon fixation rate in the oligotrophic southern station was 24% of that at the north, with more carbon fixation below the SML at 37 degreesN than at 59 degreesN. Respiration rates showed little variation with latitude, and the rates at 37 degreesN were 80% of those at 59 degreesN. Nitrate and ammonium uptake rates were very low in the oligotrophic conditions in the SML at 37 degreesN, but nitrate uptake in the euphotic zone was comparable to that at 59 degreesN. Ammonium uptake by phytoplankton was also significantly greater at 37 degreesN, in both the euphotic zone and thermocline, but uptake in the SML was only 20% of that in the SML at 59 degreesN. (C) 2001 Elsevier Science Ltd. All rights reserved.

The 1996 ‘Muslim holiday’ affair. Religious competition and state mediation in contemporary Mozambique

Political commentators often cast religious con? ict as the result of the numerical growth and political rise of a single faith. When Islam is involved, arguments about religious fundamentalism are quick to surface and often stand as an explanation in their own right. Yet, as useful as this type of explanation may be, it usually fails to address properly, if at all, two sets of important issues. It avoids, Ž rst, the question of the rise of other religions and their contribution to tensions and con? icts. Second, it reduces the role of the State to a reactive one. The State becomes an object of contest or conquest, or it is simply ignored. Adopting a different approach, this article investigates a controversy that took place in Mozambique in 1996 around the ‘ofŽ cialisation’ of two Islamic holidays. It looks at the role played by religious competition and state mediation. The article shows that the State’s abandonment of religious regulation – the establishment of a free ‘religious market’ – fostered religious competition that created tensions between faiths. It suggests that strife ensued because deregulation was almost absolute: the State did not take a clear stand in religious matters and faith organisations started to believe that the State was becoming, or could become, confessional. The conclusion discusses theoretical implications for the understanding of religious strife as well as Church and State relations. It also draws some implications for the case of Mozambique more speciŽ cally, implications which should have relevance for countries such as Malawi, Zambia and Zimbabwe where problems of a similar nature have arisen.

Annotation of Case C-272/94, Criminal proceedings against Michel Guiot and Climatec SA, ECR 1996 I-1905

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