The homeostatic chemokine CCL21 predicts mortality and may play a pathogenic role in heart failure

Background

CCL19 and CCL21, acting through CCR7, are termed homeostatic chemokines. Based on their role in concerting immunological responses and their proposed involvement in tissue remodeling, we hypothesized that these chemokines could play a pathogenic role in heart failure (HF).

Methodology/Principal Findings

Our main findings were: (i) Serum levels of CCL19 and particularly CCL21 were markedly raised in patients with chronic HF (n = 150) as compared with healthy controls (n = 20). A CCL21 level above median was independently associated with all-cause mortality. (ii) In patients with HF following acute myocardial infarction (MI; n = 232), high versus low CCL21 levels 1 month post-MI were associated with cardiovascular mortality, even after adjustment for established risk factors. (iii). Explanted failing human LV tissue (n = 29) had markedly increased expression of CCL21 as compared with non-failing myocardium (n = 5). (iv) Our studies in CCR7^−/− mice showed improved survival and attenuated increase in markers of myocardial dysfunction and wall stress in post-MI HF after 1 week, accompanied by increased myocardial expression of markers of regulatory T cells. (v) Six weeks post-MI, there was an increase in markers of myocardial dysfunction and wall stress in CCR7 deficient mice.

Conclusions/Significance

High serum levels of CCL21 are independently associated with mortality in chronic and acute post-MI HF. Our findings in CCR7 deficient mice may suggest that CCL21 is not only a marker, but also a mediator of myocardial failure. However, while short term inhibition of CCR7 may be beneficial following MI, a total lack of CCR7 during long-term follow-up could be harmful.     

Experimental evidence of density dependent activity pattern of a large herbivore in an alpine ecosystem

Density dependent processes affecting foraging strategies may in turn influence vital rates and population regulation in large herbivores. Increased competition may lower both forage availability and quality, but whether the main activity constraint at high density is increased searching time or increased digestion time is poorly investigated. In a fully replicated landscape‐scale experiment, we used long‐term data (2003–2009) from domestic sheep grazing at high and low density (80 and 25 sheep km^–2, respectively) on alpine summer ranges to test density dependence in allocation of time to feeding (moving) vs digestion (resting) activities and how this in turn affected body growth. Sheep at high density spent more time actively feeding than sheep at low density, but sheep moved shorter distances while foraging at high density. Increased activity levels at high density suggest that the main activity constraint at high density was availability of high‐quality food increasing searching time and possibly reducing intake rates. Increased movement distances at low density is consistent with a higher selection for more productive vegetation types since high‐quality patches are dispersed in the landscape. The alternative hypothesis, that food processing time increased at high density was not supported as it would have reduced overall activity levels. Individual activity levels increased body growth, but this was not sufficient to fully compensate for lower habitat quality leading to an overall reduced body growth at high density. Our experiment clearly documents changes in activity budgets and movement distances of a large herbivore at high population density, providing one potential behavioural mechanism of density dependent responses observed in vital rates.     

Antimicrobial susceptibility of Francisella noatunensis subsp. noatunensis strains isolated from Atlantic cod Gadus morhua in Norway

A total of 30 isolates of Francisella noatunensis subsp. noatunensis isolated from Atlantic cod Gadus morhua L. were tested for susceptibility, in the form of minimal inhibitory concentration (MIC) values, against the following antibacterial agents: flumequine, oxolinic acid, ciprofloxacin, florfenicol, oxytetracycline, erythromycin, streptomycin sulphate, trimetoprim/sulphadiazine and rifampin. All the isolates had a low susceptibility to oxytetracycline, trimetoprim/sulphadiazine (Tribrissen?), erythromycin, ciprofloxacin and streptomycin with MIC values of 64, 64 to 128, 16, 8 and 32 to 128 μg ml-1, respectively. The strains were, on the other hand, susceptible to florfenicol, oxolinic acid, flumequine and rifampin with MIC values of 0.5, 0.25, 0.25 and 0.25 to 1 μg ml-1, respectively.     

The importance of grazing food chain for energy flow and production in three intertidal sand bottom communities of the northern Wadden Sea

In three intertidal sand bottom communities of the Königshafen (Island of Sylt, North Sea), the biomass production and respiration of phytobenthos, phytoplankton, macrozoobenthos, and in situ community metabolism were measured monthly during 1980. The study sites were characterized by different communities (Nereis-Corophium-belt, seagrass-bed,Arenicola-flat) and by a high abundance of the molluscHydrobia ulvae. Benthic diatoms are the major constituents of plant biomass in theArenicola-flat. In this community, gross primary productivity amounts to 148 g C m^–2 a^–1. 82 % of this productivity is caused by microbenthos, whereas phytoplankton constitutes only 18 %. In the seagrass-bed, gross primary productivity amounts to 473 g C m^–2 a^–1. 79 % of this is generated by seagrass and its epiphytes, whereas microphytobenthos contributes 19 %. In theNereis-Corophium-belt, only microphytobenthos is important for biomass and primary productivity (gross: 152 g C m^–2 a^–1). Annual production of macrofauna proved to be similar in theArenicola-flat (30 g C m^–2 a^–1) to that in the seagrass-bed (29 g C m^–2 a^–1). Only one third of this amount is produced in theNereis-Corophium-belt (10 g C m^–2 a^–1). The main part of secondary production and animal respiration is contributed by grazingH. ulvae. In the seagrass-bed, 83 % of the energy used for production is obtained from the grazing food chain. In theArenicola-flat and theNereis-Corophium-belt, the importance of non-grazing species is greater. A synchrony of seasonal development of plant biomass and monthly secondary production was observed. In theArenicola-flat and the seagrass-bed, where density and production of macrofauna are high, a conspicuous decrease in biomass of microbenthos occurs during the warmer season, whereas in theNereis-Corophium-belt primary production causes an increase in microphytobenthic biomass in summer and autumn. Energy flow through the macrofauna amounts to 69 g C m^–2 a^–1 in theArenicola-flat, 85 g C m^–2 a^–1 in the seagrass-bed and 35 g C m^–2 a^–1 in theNereis-Corophium-belt. Based on the assumption that sources of food are used in proportion to their availability, 49 g C m^–2 a^–1 (Arenicola-flat), 72 g C m^–2 a^–1 (seagrass-bed) and 26 g C m^–2 a^–1 (Nereis-Corophium-belt) are estimated as taken up by the grazing food chain. All three subsystems are able to support the energy requirements from their own primary production and are not dependent on energy import from adjacent ecosystems.     

Variation in the fermentative pattern of some Saccharomyces species

Summary The acquisition of the ability to ferment galactose, maltose, sucrose, raffinose and melibiose by yeast strains of the genus Saccharomyces was investigated. During cultivation in selective media 11 strains belonging to 5 species gained the ability to ferment one or several of these sugars. De-adaptation was not usually observed after cultivation in glucose medium, indicating that the saltants are stable in this medium.     

Some factors influencing the reproduction and longevity ofCoccinella septempunctata Linnaeus [Coleoptera: Coccinellidae]

Zusammenfassung Coccinella septempunctata hat in Norwegen eine Generation pro Jahr. Die überwinterten Imagines haben in den ersten Wochen einen hohen Nahrungsbedarf (fig. 1). Das Ablegen der Eier erstreckt sich über mehrere Monate und man kann Larven fast über den ganzen Sommer finden. Die neu geschlüpften K?fer ben?tigen Nahrung, jedoch nimmt der Nahrungsbedarf nach den ersten Wochen ab. Der gesamte Nahrungsbedarf einer Population steigt w?hrend der gesamten Saison an. Die K?fer k?nnen mit künstlichen Nahrungsstoffen gefüttert werden; sie leben mehrere Monate, doch legen sie dann keine Eier ab. Gibt man ihnen jedoch in einem solchen zustand Blattl?use zu fressen, so beginnt die Eiproduktion nach etwa einer Wochen wieder. Weibchen, welche im Felde überwinterten, übertrafen die Laboratoriumstiere sowohl bezüglich der Eizahl als auch der Schlüpfprozente. Es wird die Schlussfolgerung gezogen, dassC. septempunctata geeignet ist, die Fluktuation der Blattlauspopulationen im Felde zu regulieren. Das Ausschlüpfen der K?fer sollte, damit die Blattlauspopulation klein gehalten werden kann, am besten zu einem frühen Zeitpunkt der Saison geschehen.      

The instability of physiological properties used as criteria in the taxonomy of yeasts

Summary Stock cultures of yeast strains which have acquired the ability to ferment and assimilate various sugars often represent a mixed population in regard to these features.Usually de-adaptation was not found after cultivation in glucose medium, indicating that the saltations are stable in this medium.The lyophilized cultures revived in glucose medium have the original characteristics of the strains.When single-cell colonies of various strains unable to utilize the appropriate sugar were grown in a liquid medium containing either galactose, sucrose or maltose, cells fermenting the sugar offered were found in the cultures. The acquisition of maltose-fermenting ability and the simultaneous acquisition of ability to utilize either galactose and raffinose, sucrose and raffinose, maltose and sucrose, or maltose, sucrose and -methyl-d-glucoside were observed.By an identification according to the acquired features the yeasts concerned have to be classified in other species than the original strain.     

Degradation of fully water-soluble, partially N-acetylated chitosans with lysozyme

Chitosans, prepared by homogeneous N-deacetylation of chitin, with degrees of N-acetylation ranging from 4 to 60% (F_A = 0·04 to 0·60) exhibiting full water solubility and known random distribution of acetyl groups, were degraded with lysozyme. Initial degradation rates (r) were determined from plots of the viscosity decrease (Δ1/[η]) against time of degradation. The time course of degradation of chitosans with lysozyme were non-linear, while the time course of degradation of chitosans with an oxidative-reductive depolymerization reaction (using H₂O₂) showed the expected linear relationship for a first-order, random depolymerization reaction, independent of the chemical composition of the chitosan.

The effect of lysozyme concentration and substrate concentration on the initial degradation rates were determined, showing that this lysozyme-chitosan system obeys Michaelis-Menten kinetics.

The initial degradation rates of chitosan with lysozyme increased strongly with increasing fraction of acetylated units (F_A). From a Michaelis-Menten analysis of the degradation data that assumes different catalytic activities of lysozyme for the different hexameric substrates in the polysaccharide chain, it is concluded that the hexameric substrates that contain three-four or more acetylated units contribute mostly to the initial degradation rate when lysozyme degrades partially N-acetylated chitosans.

A chitosan with a very low fraction of acetylated units (F_A = 0·010) was studied as an enzyme inhibitor. Initial degradation rates of chitosan (with different F_A values) decreased as the inhibitor concentration increased, while the relative rates stayed constant, indicating that the ratio between initial reaction rates for productive sites (hexamers containing three-four or more N-acetylated units) are unaffected by non-productive sites, as deduced from the theory of competing substrates.     

Characterization of human cytomegalovirus uracil DNA glycosylase (UL114) and its interaction with polymerase processivity factor (UL44)

Here, we report the molecular characterization of the human cytomegalovirus uracil DNA glycosylase (UNG) UL114. Purified UL114 was shown to be a DNA glycosylase, which removes uracil from double-stranded and single-stranded DNA. However, kinetic analysis has shown that viral UNG removed uracil more slowly compared with the core form of human UNG (Δ84hUNG), which has a catalytic efficiency (k_cat/K_M) 350- to 650-fold higher than that of UL114. Furthermore, UL114 showed a maximum level of DNA glycosylase activity at equimolar concentrations of the viral polymerase processivity factor UL44. Next, UL114 was coprecipitated with DNA immobilized to magnetic beads only in the presence of UL44, suggesting that UL44 facilitated the loading of UL114 on DNA. Moreover, mutant analysis demonstrated that the C-terminal part of UL44 (residues 291-433) is important for the interplay with UL114. Immunofluorescence microscopy revealed that UL44 and UL114 colocalized in numerous small punctuate foci at the immediate-early (5 and 8 hpi) phases of infection and that these foci grew in size throughout the infection. Furthermore, coimmunoprecipitation assays with cellular extracts of infected cells confirmed that UL44 associated with UL114. Finally, the nuclear concentration of UL114 was estimated to be 5- to 10-fold higher than that of UL44 in infected cells, which indicated a UL44-independent role of UL114. In summary, our data have demonstrated a catalytically inefficient viral UNG that was highly enriched in viral replication foci, thus supporting an important role of UL114 in replication rather than repair of the viral genome.