ATM engages autodegradation of the E3 ubiquitin ligase COP1 after DNA damage

The ataxia telangiectasia mutated (ATM) protein kinase is a critical component of a DNA-damage response network configured to maintain genomic integrity. The abundance of an essential downstream effecter of this pathway, the tumor suppressor protein p53, is tightly regulated by controlled degradation through COP1 and other E3 ubiquitin ligases, such as MDM2 and Pirh2; however, the signal transduction pathway that regulates the COP1-p53 axis following DNA damage remains enigmatic. We observed that in response to DNA damage, ATM phosphorylated COP1 on Ser(387) and stimulated a rapid autodegradation mechanism. Ionizing radiation triggered an ATM-dependent movement of COP1 from the nucleus to the cytoplasm, and ATM-dependent phosphorylation of COP1 on Ser(387) was both necessary and sufficient to disrupt the COP1-p53 complex and subsequently to abrogate the ubiquitination and degradation of p53. Furthermore, phosphorylation of COP1 on Ser(387) was required to permit p53 to become stabilized and to exert its tumor suppressor properties in response to DNA damage.     

Bayesian estimation of flock-level sensitivity of detection of Salmonella spp., Enteritidis and Typhimurium according to the sampling procedure in French laying-hen houses

A study was carried out to estimate the prevalence of flocks infected by Salmonella spp., S. Enteritidis and S. Typhimurium in 521 French laying-hen farms from October 1st 2004 to September 30th 2005 as part of a European Union-wide baseline study to define targets for Salmonella reduction in member states. The sampling scheme prescribed and financed by the European Commission to detect Salmonella in laying-hen flocks was based on 2 dust-samples and 5 faeces-samples per farm. A latent-class Bayesian approach for correlated tests was used to estimate the sensitivity of detection of reduced sampling schemes corresponding to the 16 combinations of 2 dust- and 5 faeces-samples. For each model the full sampling scheme (7 samples) and the reduced protocol were considered as two correlated tests, the biological principle being identical and the reduced protocol being a subset of the full sampling scheme. As the observed apparent prevalence in cage flocks was higher than in other systems (barns, outdoor, or organic) these two sub-populations were considered separately. Bayesian estimation of posterior medians with 95% probability intervals for true prevalence in cage flocks were 0.34 (0.29; 0.39) and 0.13 (0.10; 0.18) for Salmonella spp. and Salmonella Enteritidis+Typhimurium respectively. In alternative flocks posterior medians with 95% probability intervals for true prevalence were 0.09 (0.06; 0.13) and 0.05 (0.03; 0.08) for Salmonella spp. and Salmonella Enteritidis+Typhimurium, respectively. In cage flocks Bayesian estimation of posterior distributions for sensitivity indicated that at least 5 samples, including 2 dust samples were necessary to attain comparable sensitivity levels to the full sampling scheme. In alternative flocks and for Salmonella spp. 6 samples were required to ensure a comparable sensitivity level to the full sampling scheme. Detection sensitivity was improved by increasing the number of dust samples in cage farms and by increasing the total number of samples whatever their type in alternative farms.     

Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

Interleukin (IL)-10 exerts potent anti-inflammatory effects by suppression of both T-help (Th) 1 and Th2 cells. Previous studies have reported that IL-10 can ameliorate various inflammatory disorders. The present study was performed to examine whether IL-10 plasmid DNA could suppress development of atopic dermatitis (AD)-like skin lesions in NC/Nga mice, as an initial step towards the development of an appliance for use in dogs with AD. Intradermal injection of IL-10 plasmid DNA markedly inhibited the development of AD-like skin lesions, as evidenced by a marked decrease in skin symptoms and reduced inflammation within the skin lesions. Efficacy was confirmed by significant decreases in eosinophil ratio and serum IgE concentration, and a reduction in the number of Staphylococcus aureus recovered from the ear. Moreover, relative mRNA expression levels of IL-4 and interferon-γ in the skin lesions of mice injected with IL-10 plasmid DNA were also decreased compared with those of control mice. Of note, higher serum IL-10 levels in mice injected with IL-10 plasmid DNA were maintained compared with those in control mice. Taken together, the results indicate that IL-10 plasmid DNA can suppress the development of AD-like skin lesions by suppressing both Th1 and Th2 cell responses. Beneficial effects of IL-10 plasmid DNA may be expected in dogs with AD.     

Biochemical properties and microbial community structure of five different soils after atrazine addition

Atrazine is one of the most used herbicides worldwide; however, consequences of its long-term agricultural use are still unknown. A laboratory study was performed to examine changes in microbial properties following ethylamino-¹⁵N-atrazine addition, at recommended agronomic dose, to five acidic soils from Galicia (NW Spain) showing different physico-chemical characteristics, as well as atrazine application history. Net N mineralization was observed in all soils, with nitrate being the predominant substance formed. The highest values were detected in soils with low atrazine application history. From 2% to 23% of the atrazine-¹⁵N was found in the soil inorganic-N pool, the highest values being detected after 9 weeks in soils with longer atrazine application history and lower indigenous soil N mineralization. The application of atrazine slightly reduced the amount of soil N mineralized and microbial biomass at short term. Soluble carbohydrates and β-glucosidase and urease activity decreased with incubation time, but were not significantly affected by the single application of atrazine. Microbial community structure changed as consequence of both soil type and incubation time, but no changes in the phospholipid fatty acid (PLFA) pattern were detected due to recent atrazine addition at normal doses. The saturated 17- to 20-carbon fatty acids had higher relative abundance in soils with a longer atrazine history and fungal biomass, as indicated by the PLFA 18:2ω6,9, decreased with the incubation time. The results suggested that the PLFA pattern and soil N dynamics can detect the long-term impact of repeated atrazine application to agricultural soils.     

Infectiousness of pigs infected by the Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is time-dependent

The time-dependent transmission rate of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and the correlation between infectiousness, virological parameters and antibody responses of the infected pigs were studied in experimental conditions. Seven successive transmission trials involving a total of 77 specific pathogen-free piglets were carried out from 7 to 63 days post-inoculation (dpi). A semi-quantitative real time RT-PCR was developed to assess the evolution of the viral genome load in blood and nasal swabs from inoculated and contact pigs, with time. Virus genome in blood was detectable in inoculated pigs from 7 to 77 dpi, whereas viral genome shedding was detectable from nasal swabs from 2 to 48 dpi. The infectiousness of inoculated pigs, assessed from the frequency of occurrence of infected pigs in susceptible groups in each contact trial, increased from 7 to 14 dpi and then decreased slowly until 42 dpi (3, 7, 2, 1 and 0 pigs infected at 7, 14, 21, 28 and 42 dpi, respectively). These data were used to model the time-dependent infectiousness by a lognormal-like function with a latency period of 1 day and led to an estimated basic reproduction ratio, R₀ of 2.6 [1.8, 3.3]. The evolution of infectiousness was mainly correlated with the time-course of viral genome load in the blood whereas the decrease of infectiousness was strongly related to the increase in total antibodies.     

An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome

Porcine circovirus type 2 (PCV2) plays a crucial role in the pathogenesis of post-weaning multisystemic wasting syndrome (PMWS) in swine. As PCV2 displays significant homology with PCV1 (a non-pathogenic virus) at the nucleotide and amino-acid level, a discriminative antigen is needed for specific serological diagnosis. The ORF2-encoded capsid protein from PCV2 was used to develop an indirect enzyme-linked immunosorbent assay (ELISA). GST-fused capsid protein from PCV2 and GST alone (both expressed in recombinant baculovirus-infected cells) were used as antigens for serodiagnosis. The specificity of the ELISA for detection of PCV2 antibodies was demonstrated in sera from pigs experimentally infected with PCV1, PCV2 and other swine viruses. The semi-quantitative nature of the test was evaluated versus an immunoperoxidase monolayer assay (IPMA). The ELISA was performed on 322 sera from pigs in eight Brittany herds and compared with IPMA. The sensitivity (98.2%) and specificity (94.5%) of this test were considered suitable for individual serological detection. High PCV2 seroprevalence was found in sows and pigs at the end of the growth phase (18-19 weeks) in all eight herds. The seroprevalence in piglets (11-17 weeks) was statistically correlated with clinical symptoms of PMWS (93% in affected versus 54%, in non-affected farms). A cohort study performed in PMWS-free farms showed that 57% of piglets exhibited active seroconversion after 13 weeks, indicating that PCV2 infection occurred earlier in PMWS-affected piglets.     

射频-热风与热风处理保鲜白面包的比较

为将射频技术应用到袋装面包的保鲜处理中,以有效延长袋装面包的保存期,论文研究了射频-热风处理对于袋装面包的保鲜效果,并从灭菌效应、加热速率以及处理后面包的品质变化3方面与传统的热风处理方式作了对比。研究表明,与传统的热风处理相比,射频-热风处理可在更低的温度和更短的时间内获得满意的杀菌效果。对袋装切片面包进行射频-热风处理,其加热速率约为传统热风处理的30倍,并且可以更好地保持面包的品质,但其缺点是会引起面包表面积较大的收缩。     

Photo-induced living cationic polymerization of tetrahydrofuran. III. Synthesis of poly(THF-co-3-MTHF)

Poly(3-methyltetrahydrofuran)(3-MTHF) and poly(tetrahydrofuran-co-3-MTHF), having very narrow molecular weight distribution were successfully synthesized via photo-induced living cationic polymerization in the presence of diphenyliodonium hexafluorophosphate. Linear relationship between % conversion and number average molecular weight of resulting poly(3-MTHF) in the polymerization of 3-MTHF, carried out at −22°C, indicates that the 5-membered cyclic oxonium ion, being responsible for the cationic propagation is stabilized by ion pair formation with hexafluorophosphate anion, supplied from the salt. The linear relationship between two parameters, mentioned above was also observed in the copolymerization of 3-MTHF with THF, carried out at 0 and −22°C. The molecular structures including the copolymer composition and average molecular weight and its distribution is determined by reaction parameters such as monomer feed ratio and reaction temperature.     

Genetic parameters and genotype x environment interaction for feed efficiency traits in steers fed grower and finisher diets

The objective of this study was to examine the genetic parameters and genetic correlations of feed efficiency traits in steers (n = 490) fed grower or finisher diets in 2 feeding periods. A bivariate model was used to estimate phenotypic and genetic parameters using steers that received the grower and finisher diets in successive feeding periods, whereas a repeated animal model was used to estimate the permanent environmental effects. Genetic correlations between the grower-fed and finisher-fed regimens were 0.50 ± 0.48 and 0.78 ± 0.43 for residual feed intake (RFI) and G:F, respectively. The moderate genetic correlation between the 2 feeding regimens may indicate the presence of a genotype × environment interaction for RFI. Permanent environmental effects (expressed in percentage of phenotypic variance) were detected in the grower-fed steers for ADG (38%), DMI (30%), RFI (18%), and G:F (40%) and also in the finisher-fed steers for ADG (28%), DMI (35%), metabolic mid-weight (23%), and RFI (10%). Heritability estimates were 0.08 ± 0.10 and 0.14 ± 0.15 for the grower-fed steers and 0.42 ± 0.16 and 0.40 ± 17 for the finisher-fed steers for RFI and G:F, respectively. The dependency of the RFI on the feeding regimen may have serious implications when selecting animals in the beef industry. Because of the higher cost of grains, feed efficiency in the feedlot might be overemphasized, whereas efficiency in the cow herd and the backgrounding segments may have less emphasis. These results may also favor the retention (for subsequent breeding) of cows whose steers were efficient in the feedlot sector. Therefore, comprehensive feeding trials may be necessary to provide more insight into the mechanisms surrounding genotype × environment interaction in steers.     

A pmp genes-based PCR as a valuable tool for the diagnosis of avian chlamydiosis

In a previous study we described the use of a new set of PCR primers (CpsiA/CpsiB) specific of the conserved pmp-family genes of Chlamydophila abortus as an efficient tool for the detection of these bacteria in ruminants including also preliminary results on avian strains. In this work, the use of this set of primers was extended to representative strains of the six major avian serovars (serovars A-F) and to field isolates of C. psittaci. For all the studied representative strains, using purified genomic DNA as a template, CpsiA/CpsiB primers allowed, as observed for C. abortus, a minimal 10-fold PCR signal increase compared to the one observed with ompA specific primers. In comparison to primers targeting the 16S-23S rRNA intergenic spacer, similar or increased sensitivity was observed depending on the strain. All the field isolates were amplified with CpsiA/CpsiB primers. On clinical samples, our primers are the best among those tested for detection of C. psittaci by simple conventional PCR. RFLP experiments performed using PCR fragments amplified with the CpsiA/CpsiB primers gave promising results demonstrating that these primers may provide an interesting tool for molecular typing when the bacterium cannot be grown from pathological samples.