Comparison of mechanical and electrical activity and interstitial cells of Cajal in urinary bladders from wild-type and W/W-v mice

Background and purpose: W/Wv and wild-type murine bladders were studied to determine whether the W/Wv phenotype, which causes a reduction in, but not abolition of, tyrosine kinase activity, is a useful tool to study the function of bladder interstitial cells of Cajal (ICC).

Experimental approach: Immunohistochemistry, tension recordings and microelectrode recordings of membrane potential were performed on wild-type and mutant bladders.

Key results: Wild-type and W/Wv detrusors contained c-Kit- and vimentin-immunopositive cells in comparable quantities, distribution and morphology. Electrical field stimulation evoked tetrodotoxin-sensitive contractions in wild-type and W/Wv detrusor strips. Atropine reduced wild-type responses by 50% whereas a 25% reduction occurred in W/Wv strips. The atropine-insensitive component was blocked by pyridoxal-5-phosphate-6-azophenyl-2',4'-disulphonic acid in both tissue types. Wild-type and W/Wv detrusors had similar resting membrane potentials of -48 mV. Spontaneous electrical activity in both tissue types comprised action potentials and unitary potentials. Action potentials were nifedipine-sensitive whereas unitary potentials were not. Excitatory junction potentials were evoked by single pulses in both tissues. These were reduced by atropine in wild-type tissues but not in W/Wv preparations. The atropine-insensitive component was abolished by pyridoxal-5-phosphate-6-azophenyl-2',4'-disulphonic acid in both preparations.

Conclusions and implications: Bladders from W/Wv mice contain c-Kit- and vimentin-immunopositive ICC. There are similarities in the electrical and contractile properties of W/Wv and wild-type detrusors. However, significant differences were found in the pharmacology of the responses to neurogenic stimulation with an apparent up-regulation of the purinergic component. These findings indicate that the W/Wv strain may not be the best model to study ICC function in the bladder.

Design of a salisbury screen absorber using frequency selective surfaces to improve bandwidth and angular stability performance

A new design method that greatly enhances the reflectivity bandwidth and angular stability beyond what is possible with a simple Salisbury screen is described. The performance improvement is obtained from a frequency selective surface (FSS) which is sandwiched between the outermost 377 Ω/square resistive sheet and the ground plane. This is designed to generate additional reflection nulls at two predetermined frequencies by selecting the size of the two unequal length printed dipoles in each unit cell. A multiband Salisbury screen is realised by adjusting the reflection phase of the FSS to position one null above and the other below the inherent absorption band of the structure. Alternatively by incorporating resistive elements midway on the dipoles, it is shown that the three absorption bands can be merged to create a structure with a −10 dB reflectivity bandwidth which is 52% larger and relatively insensitive to incident angle compared to a classical Salisbury screen having the same thickness. CST Microwave Studio was used to optimise the reflectivity performance and simulate the radar backscatter from the structure. The numerical results are shown to be in close agreement with bistatic measurements for incident angles up to 40° over the frequency range 5.4−18 GHz.

'Like waking up in a Franz Kafka novel': Service users' experiences of the child protection system when domestic violence and acrimonious separations are involved

The challenges that arise in respect of child abuse reports made in the context of domestic violence and/or acrimonious separation have been the subject of recent academic discussion. This paper adds a service user perspective to the debate and reports on the findings from a study conducted in the Republic of Ireland. In addition to the previously established evidence about such cases, it demonstrates the level of powerlessness and frustration experienced by families who found it difficult to have their needs heard or met. It also illustrates the very detrimental emotional impact on children and parents who frequently encountered indifference as well as insensitive and gendered responses from child protection staff. The findings indicate that mainstream statutory child protection services do not have the capacity to deal with these complex cases, and advocates the adoption of alternative approaches. Importantly, the study demonstrates the necessity to pay attention to the views of service users in developing an appropriate response.

The development of national in-group bias: English children's attributions of characteristics to English, American and German people

This study investigated the development of national in-group bias in 5-11-year-old children. Three hundred and seven English children were asked to attribute characteristics to their own national group either on its own or in conjunction with attributing characteristics to one of two national out-groups, either Americans or Germans. The importance which the children ascribed to their own national identity in relationship to their other social identities was also assessed. It was found that, with increasing age, there was an increase in the number of negative characteristics attributed to the national in-group, and an increase in the number of positive characteristics attributed to the two out-groups, the net result being an overall reduction in in-group bias across this age range. However, in-group favouritism was still exhibited at all ages. Greater importance was attributed to national identity with increasing age. However, the characteristics attributed to the English in-group did not vary as a function of the comparative out-group which was present while the attributions were being made. The presence of a comparative out-group also did not affect the importance that was ascribed to the national identity. These findings suggest that children are relatively insensitive to the prevailing comparative context when making judgments about national groups.

SGK1 activity in Na+ absorbing airway epithelial cells monitored by assaying NDRG1-Thr(346/356/366) phosphorylation

Studies of HeLa cells and serum- and glucocorticoid-regulated kinase 1 (SGK1) knockout mice identified threonine residues in the n-myc downstream-regulated gene 1 protein (NDRG1-Thr(346/356/366)) that are phosphorylated by SGK1 but not by related kinases (Murray et al., Biochem J 385:1-12, 2005). We have, therefore, monitored the phosphorylation of NDRG1-Thr(346/356/366) in order to explore the changes in SGK1 activity associated with the induction and regulation of the glucocorticoid-dependent Na+ conductance (G (Na)) in human airway epithelial cells. Transient expression of active (SGK1-S422D) and inactive (SGK1-K127A) SGK1 mutants confirmed that activating SGK1 stimulates NDRG1-Thr(346/356/366) phosphorylation. Although G (Na) is negligible in hormone-deprived cells, these cells displayed basal SGK1 activity that was sensitive to LY294002, an inhibitor of 3-phosphatidylinositol phosphate kinase (PI3K). Dexamethasone (0.2 mu M) acutely activated SGK1 and the peak of this response (2-3 h) coincided with the induction of G (Na), and both responses were PI3K-dependent. While these data suggest that SGK1 might mediate the rise in G (Na), transient expression of the inactive SGK1-K127A mutant did not affect the hormonal induction of G (Na) but did suppress the activation of SGK1. Dexamethasone-treated cells grown on permeable supports formed confluent epithelial sheets that generated short circuit current due to electrogenic Na+ absorption. Forskolin and insulin both stimulated this current and the response to insulin, but not forskolin, was LY294002-sensitive and associated with the activation of SGK1. While these data suggest that SGK1 is involved in the control of G (Na), its role may be minor, which could explain why sgk1 knockout has different effects upon different tissues.

Targeted 3' processing of antisense transcripts triggers Arabidopsis FLC chromatin silencing.

Noncoding RNA is emerging as an important regulator of gene expression in many organisms. We are characterizing RNA-mediated chromatin silencing of the Arabidopsis major floral repressor gene, FLC. Through suppressor mutagenesis, we identify a requirement for CstF64 and CstF77, two conserved RNA 3'-end-processing factors, in FLC silencing. However, FLC sense transcript 3' processing is not affected in the mutants. Instead, CstF64 and CstF77 are required for 3' processing of FLC antisense transcripts. A specific RNA-binding protein directs their activity to a proximal antisense polyadenylation site. This targeted processing triggers localized histone demethylase activity and results in reduced FLC sense transcription. Targeted 3' processing of antisense transcripts may be a common mechanism triggering transcriptional silencing of the corresponding sense gene.