Correlation of Campylobacter bacteriophage with reduced presence of hosts in broiler chicken ceca

Campylobacter jejuni and Campylobacter-specific bacteriophage were enumerated from broiler chicken ceca selected from 90 United Kingdom flocks (n = 205). C. jejuni counts in the presence of bacteriophage (mean log(10) 5.1 CFU/g) were associated with a significant (P < 0.001) reduction compared to samples with Campylobacter alone (mean log(10) 6.9 CFU/g).     

Hypoxia accelerates nitric oxide-dependent inhibition of mitochondrial complex I in activated macrophages

Excess production of nitric oxide (NO) is implicated in the development of multiple organ failure, with a putative mechanism involving direct mitochondrial inhibition, predominantly affecting complex I. The persistent effects of NO on complex I may be mediated through S-nitrosylation and/or nitration. The temporal contribution of these chemical modifications to the inhibition of respiration and the influence of concurrent hypoxia have not been previously examined. We therefore addressed these questions using J774 macrophages activated by endotoxin and interferon-gamma over a 24-h period, incubated at 21% and 1% oxygen. Oxygen consumption and complex I activity fell progressively over time in the activated cells. This was largely prevented by coincubation with the nonspecific NO synthase inhibitor L-N5-(1-iminoethyl)-ornithine. Addition of glutathione ethyl ester reversed the inhibition at initial time points, suggesting an early mechanism involving nitrosylation. Thereafter, the inhibition of complex I became more persistent, coinciding with a progressive increase in mitochondrial nitration. Hypoxia accelerated the persistent inhibition of complex I, despite a reduction in the total amount of NO generated. Our results suggest that hypoxia amplified the mitochondrial inhibition induced by NO generated during inflammatory disease states.     

The docking protein FRS2alpha controls a MAP kinase-mediated negative feedback mechanism for signaling by FGF receptors

The docking protein FRS2alpha functions as a major mediator of signaling by FGF and NGF receptors. Here we demonstrate that, in addition to tyrosine phosphorylation, FRS2alpha is phosphorylated by MAP kinase on multiple threonine residues in response to FGF stimulation or by insulin, EGF, and PDGF, extracellular stimuli that do not induce tyrosine phosphorylation of FRS2alpha. Prevention of FRS2alpha threonine phosphorylation results in constitutive tyrosine phosphorylation of FRS2alpha in unstimulated cells and enhanced tyrosine phosphorylation of FRS2alpha, MAPK stimulation, cell migration, and proliferation in FGF-stimulated cells. Expression of an FRS2alpha mutant deficient in MAPK phosphorylation sites induces anchorage-independent cell growth and colony formation in soft agar. These experiments reveal a novel MAPK-mediated, negative feedback mechanism for control of signaling pathways that are dependent on FRS2 and a mechanism for heterologous control of signaling via FGF receptors.     

p53-induced inhibition of protein synthesis is independent of apoptosis.

Activation of a temperature-sensitive form of p53 in murine erythroleukaemia cells results in a rapid impairment of protein synthesis that precedes inhibition of cell proliferation and loss of cell viability by several hours. The inhibition of translation is associated with specific cleavages of polypeptide chain initiation factors eIF4GI and eIF4B, a phenomenon previously observed in cells induced to undergo apoptosis in response to other stimuli. Although caspase activity is enhanced in the cells in which p53 is activated, both the effects on translation and the cleavages of the initiation factors are completely resistant to inhibition of caspase activity. Moreover, exposure of the cells to a combination of the caspase inhibitor z-VAD.FMK and the survival factor erythropoietin prevents p53-induced cell death but does not reverse the inhibition of protein synthesis. We conclude that the p53-regulated cleavages of eIF4GI and eIF4B, as well as the overall inhibition of protein synthesis, are caspase-independent events that can be dissociated from the induction of apoptosis per se.     

UV RADIATION,PHOSPHORUS, AND THEIR COMBINED EFFECTS ON THE TAXONOMIC COMPOSITION OF PHYTOPLANKTON IN A BOREAL LAKE1

We examined how UV radiation and phosphorus (P) affect the taxonomic composition, abundance, and biomass of phytoplankton in an oligotrophic boreal lake. We exposed phytoplankton to three different solar radiation regimes (PAR + UV‐A radiation [UVAR]+ UV‐B radiation [UVBR], PAR + UVAR, and PAR only) and to five levels of P. The biomass of small chrysophytes was reduced by 350% after exposure to PAR + UVAR + UVBR compared with PAR only. No other taxa were found to be negatively affected by exposure to UVBR. Several taxa (e.g. Chry‐ sochromulina laurentiana Kling) were sensitive to UVAR, whereas others (e.g. Tabellaria flocculosa (Roth) Kutzing) were not affected by UV radiation exposure. Principal components analysis ordination separated phytoplankton that were negatively affected by UV radiation and/or positively affected by P treatments (e.g. small chrysophytes, Cryptomonas rostratiformis, T. flocculosa) from those that generally were unaffected by either treatment (e.g. desmids, some Cyanobacteria). Richness, Shannon‐Weaver diversity, and evenness were significantly higher in phytoplankton communities shielded from UVAR and UVBR. The relationship between diversity and richness was positive in all phytoplankton samples except in those exposed to UVBR. Thus, UVBR‐exposed phytoplankton communities were dominated by a few species even though the number of taxa remained relatively unchanged. Consequently, alterations in the UV environments of lakes resulting from climate warming (e.g. drought) and land‐use change (e.g. increased P export) will likely promote shifts in the community composition of lake phytoplankton.     

Seasonal plankton cycles in a temperate fjord and comments on the match-mismatch hypothesis

As a field test of the ‘match-mismatch hypothesis’(Cushing, 1972, 1975, 1990), we sampled the ichthyoplanktonand different size fractions of zooplankton at two stationsin a temperate fjord (Dabob Bay, Washington, USA) between May1985 and October 1987. We present the results of this studytogether with historical data on seasonal abundances of phytoplanktonand invertebrate predators. The peak abundance of fish larvaein late winter/early spring preceded the peak abundance of preyorganisms by an average of 3 months. Correlation analysis resultedin one negatively significant and three non-significant correlationsbetween ichthyoplankton and appropriately sized prey organisms.These field observations, showing a distinct temporal lag betweenabundances of larval fish and their zooplankton prey, do notsupport the match-mismatch hypothesis as originally formulated.We use these field data and a simulation model of the growthof young fish to discuss the role of two alternative processes—foodlimitation during late larval/juvenile stages and predation—thatmay constrain the timing of spawning of marine fish and whichmay have implications for recruitment variability as well. Thisleads us to propose that the match-mismatch hypothesis be extendedto include a ‘critical period’ that lasts longerand occurs later in the life history of marine fish than haspreviously been supposed.     

Detection of legionellae in hospital water samples by quantitative real-time LightCycler PCR

Contamination of hospital water systems with legionellae is a well-known cause of nosocomial legionellosis. We describe a new real-time LightCycler PCR assay for quantitative determination of legionellae in potable water samples. Primers that amplify both a 386-bp fragment of the 16S rRNA gene from Legionella spp. and a specifically cloned fragment of the phage lambda, added to each sample as an internal inhibitor control, were used. The amplified products were detected by use of a dual-color hybridization probe assay design and quantified with external standards composed of Legionella pneumophila genomic DNA. The PCR assay had a sensitivity of 1 fg of Legionella DNA (i.e., less than one Legionella organism) per assay and detected 44 Legionella species and serogroups. Seventy-seven water samples from three hospitals were investigated by PCR and culture. The rates of detection of legionellae were 98.7% (76 of 77) by the PCR assay and 70.1% (54 of 77) by culture; PCR inhibitors were detected in one sample. The amounts of legionellae calculated from the PCR results were associated with the CFU detected by culture (r = 0.57; P < 0.001), but PCR results were mostly higher than the culture results. Since L. pneumophila is the main cause of legionellosis, we further developed a quantitative L. pneumophila-specific PCR assay targeting the macrophage infectivity potentiator (mip) gene, which codes for an immunophilin of the FK506 binding protein family. All but one of the 16S rRNA gene PCR-positive water samples were also positive in the mip gene PCR, and the results of the two PCR assays were correlated. In conclusion, the newly developed Legionella genus-specific and L. pneumophila species-specific PCR assays proved to be valuable tools for investigation of Legionella contamination in potable water systems.