The systemic inflammatory response to exercise in adults with cystic fibrosis

AbstractExercise is associated with release of inflammatory mediators in the circulation and there is evidence that the exercising muscles and tendons are sources of interleukin-6. Due to the catabolic effects of some cytokines, increased release in circulation might contribute to alterations in body composition in adults with cystic fibrosis. We hypothesised that exercise of moderate intensity would generate increased blood concentrations of some inflammatory mediators.We investigated the change in blood concentrations of interleukin-6, tumour necrosis factor alpha and their soluble receptors after a structured exercise (box stepping) of intensity similar to that encountered during activities of daily living in 12 adults with cystic fibrosis and mean (95% confidence interval) FEV1 55.6 (44.4, 66.8)% predicted, body mass index 23.0 (21.3, 24.6) kg/m2 and 12 healthy subjects.The increments post-exercise for all inflammatory mediators and lactate corrected for the work performed until voluntary exhaustion were greater for patients, while the total work was less for patients (all p<0.01). Daytime variability of the inflammatory mediators was assessed in eight patients and was less than the change due to exercise.We report greater increments in circulating concentrations of some cytokines with moderate exercise in adults with cystic fibrosis compared to healthy subjects

Identification of Genetic Variation on the Horse Y Chromosome and the Tracing of Male Founder Lineages in Modern Breeds

<div><p>The paternally inherited Y chromosome displays the population genetic history of males. While modern domestic horses (<i>Equus caballus</i>) exhibit abundant diversity within maternally inherited mitochondrial DNA, no significant Y-chromosomal sequence diversity has been detected. We used high throughput sequencing technology to identify the first polymorphic Y-chromosomal markers useful for tracing paternal lines. The nucleotide variability of the modern horse Y chromosome is extremely low, resulting in six haplotypes (HT), all clearly distinct from the Przewalski horse (<i>E. przewalskii</i>). The most widespread HT1 is ancestral and the other five haplotypes apparently arose on the background of HT1 by mutation or gene conversion after domestication. Two haplotypes (HT2 and HT3) are widely distributed at high frequencies among modern European horse breeds. Using pedigree information, we trace the distribution of Y-haplotype diversity to particular founders. The mutation leading to HT3 occurred in the germline of the famous English Thoroughbred stallion “Eclipse” or his son or grandson and its prevalence demonstrates the influence of this popular paternal line on modern sport horse breeds. The pervasive introgression of Thoroughbred stallions during the last 200 years to refine autochthonous breeds has strongly affected the distribution of Y-chromosomal variation in modern horse breeds and has led to the replacement of autochthonous Y chromosomes. Only a few northern European breeds bear unique variants at high frequencies or fixed within but not shared among breeds. Our Y-chromosomal data complement the well established mtDNA lineages and document the male side of the genetic history of modern horse breeds and breeding practices.</p> </div

Haplotype network of the six modern and two Przewalski horse HTs.

<p>Circles represent the haplotypes with the area proportional to the observed frequency in 20 male horses in the initial Y-chromosomal sequence analysis (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060015#pone.0060015.s014" target="_blank">Table S4</a>). HT1, n = 7 (three Lipizzan, two Arabian, one Shetland pony, one Shire horse); HT2, n = 5 (five Lipizzan); HT3, n = 3 (one Thoroughbred, one Trakehner, one Quarter horse); HT 4 (one Icelandic horse), HT5 (one Norwegian Fjord horse), HT6 (one Shetland pony), HTPrz1 (one Przewalski horse), HTPrz2 (one Przewalski horse). A dashed line between the haplotypes indicates, that the polymorphism is located on the highly variable contig YE3, which was omitted when estimating divergence time and nucleotide diversity.</p

Geographic distribution and history of Y-haplotypes in modern horse breeds.

<p>(a) Geographic distribution of Y-chromosomal haplotypes in a set of modern horse breeds. Only a few important breeds are specified, the full list with information on breeds and HT frequencies is given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060015#pone.0060015.s017" target="_blank">Table S7</a>. (b) Origin of modern domestic horse founders deduced from pedigree data. Each founder is represented by a drum with its size proportionally to the number of offspring in the dataset. The number in the drums serve as founder identifiers. Detailed information on founders (name, year of birth, breed, origin, information on import) is listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060015#pone-0060015-t001" target="_blank">Table 1</a>. (c) Male introgression routes deduced from the pedigree and the distribution of HT2 and HT3 in our dataset. HT2 (yellow arrows) arrived from South-East at early times and has been spread during the Neapolitan and Oriental introgression waves, but did not reach Northern Europe and the Iberian peninsula. The English wave in red is well documented through pedigree data and the spread of HT3 (red arrows). Due to the ubiquitous occurrence of HT1, this haplotype is not considered. The black solid lines reflect the limits of the observation of HT2 and HT3.</p

Pedigree of Darley Arabians progeny depicting the origin of HT3 from HT2.

<p>Breeds of analysed males are listed on the bottom and the haplotypes of their ancestors are reconstructed (HT2-yellow, HT3-red, unknown-grey). Selected famous stallions are shown by name; dotted lines connect relatives where at least one ancestor is omitted. No descendants from “Pot8os” and “Waxy” were available apart from “Whalebone, 1807”. The mutation leading to HT3 must have occurred either in the germline of stallion “Eclipse” <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060015#pone.0060015-Eclipse1" target="_blank">[54]</a> or in his son “Pot8os” or in his grandson “Waxy” and rose to very high frequency in the English Thoroughbred and many sport horse breeds through the progeny of the stallion “Whalebone”.</p

Data_file_Russian_Reindeer

Whole-genome analysis of four female samples of Rangifer tarandus inhabiting Russia (Suppl_Bovine_Russian_Reindeer and Suppl_Ovine_Russian_Reindeer): 1-column -Chromosome; 2 column - SNP; 3- column –Control; 4 column-Allele1; 5 column –Allele2; 6 column -Number of homozygous animals1; 7 column -Number of heterozygous animals; 8 column -Number of homozygous animals2; 9 column -Observed heterozygosity; 10 column- Expected heterozygosity; 11-Hardy-Weinberg equilibrium; 12-Number of animals

Specificity and time dependence of the biological activity of r28M.

<p>(A) CSPG4 positive (IPC-298) or negative cells (U-251 MG) were incubated together with PBMC and 1000 ng/ml enriched r28M fraction in a final volume of 150 μl for 24, 48 as well as 72 hrs to measure tumor cell viability. Results show samples normalized to the controls. Cells incubated without r28M served as controls and their number was considered as 1. Values are means ± SE. Significance levels: x: p ≤ 0.05; xx: p ≤ 0.01; xxx: p ≤ 0.001. (B) Plates were coated with the indicated antibodies or PBS and incubated with PBMC for 30 min, 2 hrs, 8 hrs as well as 24 hrs, respectively. Preparations were pipetted in triplicates. The supernatants of these triplicates were pooled and the IL-2 concentrations were measured by ELISA.</p

Impact of detergents on the r28M activity.

<p>The effects of the addition of the detergents Tween® 20 (A), Tween® 80 (B), saponin (C) and Na-deoxycholate (D) on the efficacy of PBMC to kill CSPG4 positive tumor cells (IPC-298) induced by 1000 ng/ml enriched r28M fraction are shown. Full grey bars: considered concentration of detergents (no positive or negative influence on the growth rate of CSPG4 positive cells and/or PBMC—see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0140471#pone.0140471.s002" target="_blank">S2 Fig</a>). Dashed bars: concentration of detergents that influenced cell growth either positively or negatively (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0140471#pone.0140471.s002" target="_blank">S2 Fig</a>); therefore data was not included in the given statistical analysis. Results show samples normalized to the controls. Cells incubated without r28M served as controls, their number was considered as 1. Values are means ± SE. Significance levels: x: p ≤ 0.05; xx: p ≤ 0.01; xxx: p ≤ 0.001.</p

Optimized purification of r28M from plasma from transgenic cattle.

<p>The r28M (A) as well as BSA contents (B) of plasma after precipitation with ammonium sulfate (AS) or polyethylene glycol (PEG) were determined by competitive ELISAs and depicted as percentages of the bovine plasma feedstock used for purification of r28M. One representative example out of four independent experiments is shown. (C) The flow-through (FT) and eluate fraction (E) of plasma purified via Protein A or G on two different gels are shown. 500 ng of each fraction were separated by a 8% SDS-PAGE under non-reducing conditions and subsequently silver-stained. Cut FT and E sections of Protein A were on the same gel, but not on nearby slots. One representative example out of three independent experiments is shown.</p