Elevated expression of both mRNA and protein levels of IL-17A in sputum of stable Cystic Fibrosis patients

Background

T helper 17 (Th17) cells can recruit neutrophils to inflammatory sites through production of IL-17, which induces chemokine release. IL-23 is an important inducer of IL-17 and IL-22 production. Our aim was to study the role of Th17 cells in cystic fibrosis (CF) lung disease by measuring IL-17 protein and mRNA levels and IL-22 and IL-23 mRNA in sputum of clinically stable CF patients and by comparing these levels with healthy controls.

Methods

Sputum induction was performed in adult CF patients outside of an exacerbation and healthy control subjects. IL-17A protein levels were measured in supernatants with cytometric bead array (CBA) and RNA was isolated and quantitative RT-PCR was performed for IL-17A, IL-22 and IL-23.

Results

We found significantly higher levels of IL-17A protein and mRNA levels (both: p < 0.0001) and IL-23 mRNA levels (p < 0.0001) in the sputum of CF group as compared to controls. We found very low levels of IL-22 mRNA in the CF group. The levels of IL-17 and IL-23 mRNA were higher in patients chronically infected with Pseudomonas aeruginosa (P. aeruginosa) as compared to those who were not chronically infected with P. aeruginosa. The presence of Staphylococcus aureus (S. aureus) on sputum did not affect the IL-17 or IL-23 levels. There was no correlation between IL-17 or IL-23 levels and FEV₁ nor sputum neutrophilia.

Conclusion

The elevated levels of IL-17 and IL-23 might indicate that Th17 cells are implicated in the persistent neutrophil infiltration in CF lung disease and chronic infection with P. aeruginosa.     

A novel aux/IAA28 signaling cascade activates GATA23-dependent specification of lateral root founder cell identity

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Highlights? GATA23 is involved in lateral root founder cell specification ? A coordinated migration of pericycle nuclei precedes the first asymmetric division ? An Aux/IAA28-dependent auxin signaling cascade controls GATA23 expression     

Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells

The ability to rapidly and efficiently generate reliable Cre/loxP conditional transgenic mice would greatly complement global high-throughput gene targeting initiatives aimed at identifying gene function in the mouse. We report here the generation of Cre/loxP conditional ROSA26-targeted ES cells within 3–4 weeks by using Gateway® cloning to build the target vectors. The cDNA of the gene of interest can be expressed either directly by the ROSA26 promoter providing a moderate level of expression or by a CAGG promoter placed in the ROSA26 locus providing higher transgene expression. Utilization of F1 hybrid ES cells with exceptional developmental potential allows the production of germ line transmitting, fully or highly ES cell-derived mice by aggregation of cells with diploid embryos. The presented streamlined procedures accelerate the examination of phenotypical consequences of transgene expression. It also provides a unique tool for comparing the biological activity of polymorphic or splice variants of a gene, or products of different genes functioning in the same or parallel pathways in an overlapping manner.     

The phylogeny of the genus Nitrobacter based on comparative rep-PCR, 16S rRNA and nitrite oxidoreductase gene sequence analysis

Strains of Nitrobacter mediate the second step in the nitrification process by oxidizing nitrite to nitrate. The phylogenetic diversity of the genus is currently not well investigated. In this study, a rep-PCR profile and the nearly complete 16S rRNA gene sequence of 30 strains, comprising a wide physiological as well as ecological diversity and encompassing representatives of the four species, were determined. The sequence diversity of the 16S rRNA gene between different species was low, indicating the need for additional phylogenetic markers. Therefore, primers were developed for amplifying the complete nxrX gene and a 380bp fragment of the nxrB1 gene, which are both genes involved in the nitrite oxidation process. These genes confirmed the division into phylogenetic groups revealed by the 16S rRNA gene but showed a better discriminatory power. They can be a valuable additional tool for phylogenetic analysis within the genus Nitrobacter and can assist in the identification of new Nitrobacter isolates.     

An omnibus consistent adaptive percentile modified Wilcoxon rank sum test with applications in gene expression studies

Summary We present an adaptive percentile modified Wilcoxon rank sum test for the two‐sample problem. The test is basically a Wilcoxon rank sum test applied on a fraction of the sample observations, and the fraction is adaptively determined by the sample observations. Most of the theory is developed under a location‐shift model, but we demonstrate that the test is also meaningful for testing against more general alternatives. The test may be particularly useful for the analysis of massive datasets in which quasi‐automatic hypothesis testing is required. We investigate the power characteristics of the new test in a simulation study, and we apply the test to a microarray experiment on colorectal cancer. These empirical studies demonstrate that the new test has good overall power and that it succeeds better in finding differentially expressed genes as compared to other popular tests. We conclude that the new nonparametric test is widely applicable and that its power is comparable to the power of the Baumgartner‐Weiß‐Schindler test.     

Comparative and functional morphology of the buccal cavity of Diplogastrina (Nematoda) and a first outline of the phylogeny of this taxon*

The Diplogastrina include about 290 species of free living nematodes. Traditional classifications of this taxon are not based upon hypotheses of phylogenetic relationships. The highly variable structures of the buccal cavity were examined in 21 species using light microscopy and SEM. The function of the stomatal structures was studied with the aid of video recordings of living worms. The morphological data were used to reconstruct a first outline of the phylogenetic relationships of the Dipolgastrina. A rhabditoid gymnostomatal tube which is longer than wide, a short stegostom and a small dorsal tooth as in Pseudodiplogasteroides belong to the stem species pattern of Diplogastrina. Diplogastrina with a ' Rhabditis '-like gymnostomatal tube feed on bacteria and small fungal spores. A short and broad gymnostom as well as a right subventral tooth which forms a functional unit with the dorsal tooth were acquired step by step in the ancestral line leading to Mononchoides and Tylopharynx . The cuticularized cheilostom was divided into six plates connected by pliable regions twice independently within the Diplogastrina. The teeth-bearing posterior part of the buccal capsule can move forewards by pushing apart the plates of the cheilostom so that the teeth can get in contact with food items that are too big to be sucked into the buccal cavity. Diplogastrina with a divided cheilostom can feed not only on bacteria, but also on larger fungal spores, ciliates or other nematodes. Tylopharynx is specialized to rip apart the cell wall of fungal hyphae with the movements of a dorsal and a subventral tooth in order to suck out the contents of the fungus. This shows that the transformation of the buccal cavity in Diplogastrina is linked with an expansion of ecological niches.     

The use of functional MRI to evaluate cervical flexor activity during different cervical flexion exercises.

The purpose of this study was to investigate the recruitment pattern of deep and superficial neck flexors evoked by three different cervical flexion exercises using muscle functional MRI. In 19 healthy participants, transverse relaxation time (T2) values were calculated for the longus colli (Lco), longus capitis (Lca), and sternocleidomastoid (SCM) at rest and following three exercises: conventional cervical flexion (CF), craniocervical flexion (CCF), and a combined craniocervical flexion and cervical flexion (CCF-CF). CCF-CF gave the highest T2 increase for all muscles. CCF displayed a significantly higher T2 increase for the Lca compared with the Lco and the SCM. When comparing the CCF and CF, no significant difference was found for the Lca, whereas the Lco and SCM displayed a higher T2 increase during CF compared with CCF. This study shows that muscle functional MRI can be used to characterize the specific activation levels and recruitment patterns of the superficial and deep neck flexors during different cervical flexion exercises. During CCF-CF, all synergists are maximally recruited, which makes this exercise useful for high-load training. CCF may provide a more specific method to assess and retrain Lca muscle performance compared with CF and CCF-CF. This study highlights the need to differentiate between the Lco and Lca when evaluating their function, since these results demonstrate a clear difference in activation of both muscles.