The nucleotide composition of microsatellites impacts both replication fidelity and mismatch repair in human colorectal cells

Microsatellite instability is a key mechanism of colon carcinogenesis. We have previously studied mutations within a (CA)13 microsatellite using an enhanced green fluorescent protein (EGFP)-based reporter assay that allows the distinction of replication errors and mismatch repair (MMR) activity. Here we utilize this assay to compare mutations of mono- and dinucleotide repeats in human colorectal cells. HCT116 and HCT116+chr3 cells were stably transfected with EGFP-based plasmids harboring A10, G10, G16, (CA)13 and (CA)26 repeats. EGFP-positive mutant fractions were quantitated by flow cytometry, mutation rates were calculated and the mutant spectrum was analyzed by cycle sequencing. EGFP fluorescence pattern changed with the microsatellite's nucleotide sequence and cell type and clonal variations were observed in mononucleotide repeats. Replication errors (as calculated in HCT116) at A10 repeats were 5–10-fold higher than in G10, G16 were 30-fold higher than G10 and (CA)26 were 10-fold higher than (CA)13. The mutation rates in hMLH1-proficient HCT116+chr3 were 30–230-fold lower than in HCT116. MMR was more efficient in G16 than in A10 clones leading to a higher stability of poly-G tracts. Mutation spectra revealed predominantly 1-unit deletions in A10, (CA)13 and G10 and 2-unit deletions or 1-unit insertion in (CA)26. These findings indicate that both replication fidelity and MMR are affected by the microsatellite's nucleotide composition

Repeatability of strongyle egg counts in naturally infected horses

The selective treatment of horses is used to decrease the number of anthelmintic treatments by only treating a proportion of animals in the population. One way to select animals for treatment is to identify low and high egg-shedders using faecal egg counts (FEC); then to treat only the high egg-shedders. The value of this method is enhanced if differences among individuals in the level of egg-shedding remain consistent over time. One way to assess the stability of the rankings of animals over time is to measure the repeatability which is defined as the variance between horses divided by the total variance. The repeatability varies between 0 (no consistency in the values) to 1 (perfect consistency). To determine the repeatability of egg-shedding in naturally infected horses over time, 2637 FEC and raw egg counts (REC; i.e. originally counted eggs without multiplication factor) from 303 horses were analysed. The distribution of FEC was more overdispersed than a Poisson distribution. Therefore, a negative-binomial model was used. The within-horse-repeatability of RECs was 0.52. In a second analysis, we excluded horses that were treated with anthelmintic drugs during the study by eliminating all REC within the egg-reappearance-period. Here, the within-horse-repeatability was very similar at 0.53. The results show that egg-shedding of individual horses stays fairly consistent over time. They also show that animals which shed relatively high numbers of nematode eggs can be identified and targeted for treatment

Copeptin serum levels and 30 day survival.

<p>Serum levels of copeptin in 30-day survivors and non-survivors in the total cohort (A), in medical patients (B) and in patients that were admitted because of cardiac surgery or heart valve intervention (C); outliers not shown; * p<0.05; ¶ p<0.005</p

Survival according to tertiles of copeptin.

<p>Survival according to tertiles of copeptin in the total cohort (A), in medical patients (B) and in patients that were admitted due to cardiac surgery or heart valve intervention (C);</p

Clinical and demographic baseline characteristics of the study population.

<p>Clinical and demographic baseline characteristics of the study population.</p