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51.
The innate immune system plays a central role in host defence against viruses. While many studies portray mechanisms in early antiviral immune responses of humans and mice, much remains to be discovered about these mechanisms in the cat. With the objective of shedding light on early host-virus interactions in felids, we have developed 12 real-time TaqMan(?) qPCR systems for feline genes relevant to innate responses to viral infection, including those encoding for various IFNα and IFNω subtypes, IFNβ, intracellular antiviral factor Mx, NK cell stimulator IL-15 and effectors perforin and granzyme B, as well as Toll-like receptors (TLRs) 3 and 8. Using these newly developed assays and others previously described, we measured the relative expression of selected markers at early time points after viral infection in vitro and in vivo. Feline embryonic fibroblasts (FEA) inoculated with feline leukemia virus (FeLV) indicated peak levels of IFNα, IFNβ and Mx expression already 6h after infection. In contrast, Crandell-Rees feline kidney (CrFK) cells inoculated with feline herpes virus (FHV) responded to infection with high levels of IFNα and IFNβ only after 24h, and no induction of Mx could be detected. In feline PBMCs challenged in vitro with feline immunodeficiency virus (FIV), maximal expression levels of IFNα, β and ω subtype genes as well as IL-15 and TLRs 3, 7 and 8 were measured between 12 and 24h after infection, whereas expression levels of proinflammatory cytokine gene IL-6 were consistently downregulated until 48h post inoculation. A marginal upregulation of granzyme B was also observed within 3h after infection. In an in vivo experiment, cats challenged with FIV exhibited a 2.4-fold increase in IFNα expression in blood 1 week post infection. We furthermore demonstrate the possibility of stimulating feline immune cells in vitro with various immune response modifiers (IRMs) already known for their immunostimulatory properties in mice and humans, namely Poly IC, Resiquimod (R-848) and dSLIM?, a synthetic oligonucleotide containing several unmethylated CpG motifs. Stimulation of feline PBMCs with dSLIM? and R-848 effectively enhanced expression of IFNα within 12h by factors of 6 and 12, respectively, and Poly IC induced an increase in Mx mRNA expression of 28-fold. Altogether, we describe new molecular tools and their successful use for the characterization of innate immune responses against viruses in the cat and provide evidence that feline cells can be stimulated by synthetic molecules to enhance their antiviral defence mechanisms.  相似文献   
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This greenhouse study aimed to analyze the impact of arbuscular mycorrhizal (AM) fungal associations on maize (Zea mays L. hybrid Pioneer 3905) in order to compare their functional compatibility and efficiency. The AM fungus species used for this study were Glomus aggregatum, G. etunicatum, G. mosseae, and G. versiforme. Shoot and leaf masses, chlorophyll, soluble protein, total and reducing sugar, carbon (C), and nitrogen (N) concentrations, and glutamine synthetase (GS) activity in the maize leaves were analyzed. The root colonization ranged from 26% to 72% depending on the AM fungus species. Leaf mass was significantly higher when maize plants were colonized with G. etunicatum in comparison to the non‐AM control. The mycorrhizal effect on dry leaf mass ranged from 15.9% to 23.9% depending on the AM species. However, the total shoot mass did not differ significantly among the treatments. The mycorrhizal treatment had a marginally significant effect on the chlorophyll concentrations in maize leaves. The protein concentration was the highest in the plants colonized with G. etunicatum and the N percentage was significantly higher in the leaves of plants colonized by G. versiforme or G. aggregatum than those with G. mosseae. However, the AM colonization did not significantly alter the GS activity among the treatments. The highest sugar concentrations were detected in the leaves of plants colonized by G. versiforme. The sugar concentrations as well as the C percentage were lower in the leaves of plants colonized by G. etunicatum compared to the other mycorrhizal treatments but the values were comparable to the non‐AM control. Our overall results suggest that the expression of the mycorrhizal potential in the maize host plants varies among AM fungal species.  相似文献   
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Genetic maps published for elite durum wheat (Triticum turgidum durum Desf.), were mainly related to Italian and North American pools. To provide additional information dedicated to durum elite pool, a recombinant inbred population (217 RILs) derived form 2 French commercial varieties (Ixos and Primadur), has been mapped with 529 loci (98 AFLPs, 67 SSRs, 1 ISBP and 363 DArTs). Genetic map spanned a total of 2,082 cM (5.7 cM/marker on average). Recombination rate variation over the genome was documented through the analysis of six segregating populations representing a total of 48 RILs × 6, issued from an half diallel design based on four elite lines—Neodur, Ixos, Lloyd and Primadur—including the 2 previous genitors. Each set of three genetically connected populations (48 RILs × 3) was used to build the consensus parental maps for each genitor. To construct the four consensus parental maps, marker ordering was determined on 217 RILs map which was an extended population of the most polymorphic cross (Ixos × Primadur). Sizeable, recombination rate variation has been observed between the four parental maps both at the marker pair scale and at the linkage group scale. Two of the parents, Lloyd and Primadur, had strong, opposite effects increasing and decreasing, respectively the recombination rate. Additional studies are proposed to increase our understanding of this variation, with the identification of environmental or genetic factors affecting recombination rate.  相似文献   
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Microbial communities in soil A horizons derive their carbon from several potential sources: organic carbon (C) transported down from overlying litter and organic horizons, root-derived C, or soil organic matter. We took advantage of a multi-year experiment that manipulated the 14C isotope signature of surface leaf litter inputs in a temperate forest at the Oak Ridge Reservation, Tennessee, USA, to quantify the contribution of recent leaf litter C to microbial respiration and biomarkers in the underlying mineral soil. We observed no measurable difference (<∼40‰ given our current analytical methods) in the radiocarbon signatures of microbial phospholipid fatty acids (PLFA) isolated from the top 10 cm of mineral soil in plots that experienced 3 years of litterfall that differed in each year by ∼750‰ between high-14C and low-14C treatments. Assuming any difference in 14C between the high- and low-14C plots would reflect C derived from these manipulated litter additions, we estimate that <∼6% of the microbial C after 4 years was derived from the added 1-4-year-old surface litter. Large contributions of C from litter < 1 year (or >4 years) old (which fell after (or prior to) the manipulation and therefore did not differ between plots) are not supported because the 14C signatures of the PLFA compounds (averaging 200-220‰) is much higher that of the 2004-5 leaf litter (115‰) or pre-2000 litter. A mesocosm experiment further demonstrated that C leached from 14C-enriched surface litter or the O horizon was not a detectable C source in underlying mineral soil microbes during the first eight months after litter addition. Instead a decline in the 14C of PLFA over the mesocosm experiment likely reflected the loss of a pre-existing substrate not associated with added leaf litter. Measured PLFA Δ14C signatures were higher than those measured in bulk mineral soil organic matter in our experiments, but fell within the range of 14C values measured in mineral soil roots. Together, our experiments suggest that root-derived C is the major (>60%) source of C for microbes in these temperate deciduous forest soils.  相似文献   
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Two layouts for microtitre plates, which should serve for in-vitro susceptibility testing in routine diagnostics, have been set up by the working group "Antibiotic resistance" of the German Society for Veterinary Medicine. One of these layouts was designed for the testing of bacteria from cases of mastitis and the other for bacteria from infections in large food-producing animals. The choice of the antimicrobial agents and their concentrations to be included in these layouts were based on (1) the bacteria frequently associated with the respective diseases/animals, (2) the antimicrobial agents licensed for therapeutic use in these diseases/animals, (3) the currently available breakpoints, and (4) cross-resistances between the antimicrobial agerts so far known to occur in the respective bacteria.  相似文献   
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Despite the numerous available possibilities for the surgical treatment of peripheral nerve lesions found in the dog, the success of these treatments is often unsatisfactory. It has been proven that Schwann cells (SC) have a positive influence on the regeneration of nerve stumps. Implanting a guidance channel seeded with autologous SC at the lesion site could be a new therapeutic approach. The aim of this research was to investigate the in vitro cultivation and expansion of canine SC as the main requirement for the treatment referred to above. Biopsies were carried out on 17 nerve samples originating from dogs of different breed, age, gender and condition. The reexplantation method was employed, followed by dissociation using hyaluronidase, collagenase and trypsin and further expansion. The samples were divided into six groups which were treated with a varying combination of mitogens (forskolin, bovine PEX, choleratoxin, heregulin). To obtain the quantities of SC, the specimens were immunostained by a p75-antibody. By employing a growing number of agents it was possible to obtain an increase in both the quantity of cells and purity of cultures. A maximum of 16x10(5) cells per millilitre of suspension was achieved. The largest SC purity measured 27.1%. The maximum SC quantity achieved was 43.3x10(4) SC per millilitre.  相似文献   
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