DNA methylation patterns of Brachypodium distachyon chromosomes and their alteration by 5-azacytidine treatment

Sequential immunolocalisation of 5-methylcytosine (5-MeC) and fluorescence in situ hybridisation with chromosome-specific BAC clones were performed on Brachypodium distachyon mitotic metaphase chromosomes to determine specific DNA methylation patterns of each chromosome in the complement. In the majority of cells examined, chromosomes Bd4 and Bd5, which bear the loci of 5S and 35S ribosomal DNA, respectively, had characteristic 5-MeC patterns. In contrast, the distribution of 5-MeC along the metacentric chromosome pairs Bd1, Bd2 and Bd3 was more variable. There were numerous differences in distribution of methylated sites between homologous chromosomes as well as between chromosome arms. Some chromosome sites, such as pericentromeric regions, were highly methylated in all chromosomes. Additionally, the influence of a hypomethylating agent, 5-azacytidine, on B. distachyon chromosome methylation patterns was confirmed. It was found that some chromosome pairs underwent demethylation more easily than others, but there was no apparent regularity in demethylation of particular chromosome segments

Relationship Between Anti-DFS70 Autoantibodies and Oxidative Stress

Background: The anti-DFS70 autoantibodies are one of the most commonly and widely described agent of unknown clinical significance, frequently detected in healthy individuals. It is not known whether the DFS70 autoantibodies are protective or pathogenic. One of the factors suspected of inducing the formation of anti-DFS70 antibodies is increased oxidative stress. We evaluated the coexistence of anti-DFS70 antibodies with selected markers of oxidative stress and investigated whether these antibodies could be considered as indirect markers of oxidative stress. Methods: The intensity of oxidative stress was measured in all samples via indices of free-radical damage to lipids and proteins such as total oxidant status (TOS), concentrations of lipid hydroperoxides (LPH), lipofuscin (LPS), and malondialdehyde (MDA). The parameters of the non-enzymatic antioxidant system, such as total antioxidant status (TAS) and uric acid concentration (UA), were also measured, as well as the activity of superoxide dismutase (SOD). Based on TOS and TAS values, the oxidative stress index (OSI) was calculated. All samples were also tested with indirect immunofluorescence assay (IFA) and 357 samples were selected for direct monospecific anti DFS70 enzyme-linked immunosorbent assay (ELISA) testing. Results: The anti-DFS70 antibodies were confirmed by ELISA test in 21.29% of samples. Compared with anti-DFS70 negative samples we observed 23% lower concentration of LPH (P =.038) and 11% lower concentration of UA (P =.005). TOS was 20% lower (P =.014). The activity of SOD was up to 5% higher (P =.037). The Pearson correlation showed weak negative correlation for LPH, UA, and TOS and a weak positive correlation for SOD activity. Conclusion: In samples positive for the anti-DFS70 antibody a decreased level of oxidative stress was observed, especially in the case of samples with a high antibody titer. Anti-DFS70 antibodies can be considered as an indirect marker of reduced oxidative stress or a marker indicating the recent intensification of antioxidant processes

Highly multiplexed quantitative PCR-based platform for evaluation of chicken immune responses

To address the need for sensitive high-throughput assays to analyse avian innate and adaptive immune responses, we developed and validated a highly multiplexed qPCR 96.96 Fluidigm Dynamic Array to analyse the transcription of chicken immune-related genes. This microfluidic system permits the simultaneous analysis of expression of 96 transcripts in 96 samples in 6 nanolitre reactions and the 9,216 reactions are ready for interpretation immediately. A panel of 89 genes was selected from an RNA-seq analysis of the transcriptional response of chicken macrophages, dendritic cells and heterophils to agonists of innate immunity and from published transcriptome data. Assays were confirmed to be highly specific by amplicon sequencing and melting curve analysis and the reverse transcription and preamplification steps were optimised. The array was applied to RNA of various tissues from a commercial line of broiler chickens housed at two different levels of biosecurity. Gut-associated lymphoid tissues, bursa, spleen and peripheral blood leukocytes were isolated and transcript levels for immune-related genes were defined. The results identified blood cells as a potentially reliable indicator of immune responses among all the tissues tested with the highest number of genes significantly differentially transcribed between birds housed under varying biosecurity levels. Conventional qPCR analysis of three differentially transcribed genes confirmed the results from the multiplex qPCR array. A highly multiplexed qPCR-based platform for evaluation of chicken immune responses has been optimised and validated using samples from commercial chickens. Apart from applications in selective breeding programmes, the array could be used to analyse the complex interplay between the avian immune system and pathogens by including pathogen-specific probes, to screen vaccine responses, and as a predictive tool for immune robustness

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

Effect of intensity of enzymatic darkening on formation of impact black spot in advanced breeding material of potato after long-term storage

Celem podjętych badań było określenie wpływu temperatury przechowywania nowych genotypów ziemniaka na intensywność ciemnej plamistości pouderzeniowej. Materiałem badawczym było 178 genotypów ziemniaka trzech grup wczesności. Materiał hodowlany uprawiano na polu doświadczalnym IHAR Oddział Jadwisin. Po zbiorze i po przygotowawczym przechowywaniu bulwy umieszczono w doświadczalnej przechowalni w dwóch temperaturach 80C i 50C, przy wilgotności względnej powietrza 90-95%. Doświadczenie prowadzono przez pięć sezonów przechowalniczych. Podatność na ciemną plamistość pouderzeniową oraz ciemnienie enzymatyczne genotypów oznaczano w dwóch terminach: bezpośrednio po zbiorze oraz po 7 miesiącach przechowywania. Na podstawie przeprowadzonych badań wykazano, że temperatura przechowywania zaawansowanych materiałów hodowlanych ziemniaka miała istotny wpływ na powstawanie ciemnej plamistości pouderzeniowej (CPP) oraz intensywność ciemnienia enzymatycznego bulw. Istnieje istotna współzależność cech pomiędzy intensywnością ciemnienia enzymatycznego, a podatnością genotypów ziemniaka mało wrażliwego i wrażliwego na ciemną plamistość pouderzenio-wą przechowywanych w niskiej temperaturze. Takich zależności nie obserwowano w genotypach ziemniaka ocenianych bezpośrednio po zbiorze oraz składowanych w wyższej temperaturze. Najniższy współczynnik zbieżności (φ2 = 36%) otrzymano dla genotypów mało wrażliwych na powstawanie CPP przechowywanych w niskiej temperaturze powietrza. Dla takich genotypów intensywność ciemnienia enzymatycznego może być szybkim, subiektywnym wskaźnikiem oceny podatności bulw ziemniaka na powstawianie CPP.The aim of this study was to determine the effect of storage temperature of new potato genotypes on the intensity of black spot. The study was conducted on 178 potato genotypes representing three groups of earliness. The breeding material was grown on the experimental field of IHAR Jadwisin Division. After harvest (third decade of September ) tubers were placed in an experimental storage facility at two temperatures, 8°C and 5°C, at relative humidity of 90-95%. The experiment was conducted in five storage seasons, in the years 2008-2012. Susceptibility of the genotypes to black spot was determined in two periods: immediately after harvest and after 7 months of storage at two temperatures. On the basis of the studies it was demonstrated that storage temperature of potato breeding material had a significant impact on blackspot formation and enzymatic browning intensity. There is a significant correlation between the intensity of enzymatic darkening and the susceptibility of potato genotypes low sensitive and sensitive to blackspot stored at low temperature. Such a relationship was not observed in potato genotypes directly after harvest and stored at higher temperature. The lowest rate of convergence (φ2 = 36%) was obtained for genotypes low sensitive to the formation of impact blackspot stored at low temperature. For these genotypes enzymatic darkening intensity may be a subjective indicator for the assessment the susceptibility of potatoes to the formation of impact black spot

Air pollution and respiratory health of children: the PEACE panel study in Katowice, Poland.

This study was carried out within the framework of the multicentre Pollution Effects on Asthmatic Children in Europe (PEACE) project. Two panels of mildly asthmatic children were studied. Seventy two children living in the Upper Silesia (the largest Polish industrial agglomeration) and 73 children in the control panel were followed up during two winter months in 1994. Ambient concentration of particles with a 50% cut-off aerodynamic diameter of 10 μm (PM10) black smoke, SO2 and NO2 were measured and peak respiratory flows and respiratory symptoms were recorded on a daily basis. There were no substantial differences in exposure to air pollution and the prevalence of respiratory symptoms between the urban and the control panels. No severe smog episodes were observed. No consistent association between daily changes of air pollution levels and daily variations of peak expiratory flows or respiratory symptoms prevalence and incidence was found. In conclusion, no clear effect of air pollution on respiratory health could be observed