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.

Anatomy and facies association of a drumlin in Co. Down, Northern Ireland, from seismic and electrical resistivity surveys

Seismic refraction and electrical resistivity geophysical techniques were used to reconstruct the internal architecture of a drumlin in Co. Down, Northern Ireland. Geophysical results were both validated and complemented by borehole drilling, ground water flow modelling, and geologic mapping. The geophysical anatomy of the drumlin consists of five successive layers with depth including; topsoil, partially saturated and saturated glacial tills, and weathered and more competent greywacke bedrock. There are numerous, often extensive inclusions of clay, sand, gravel, cobbles, and boulders within the topsoil and the till units. Together geophysical and geotechnical findings imply that the drumlin is part of the subglacial lodgement, melt-out, debris flow, sheet flow facies described by previous authors, and formed by re-sedimentation and streamlining of pre-existing sediments during deglaciation of the Late Devensian ice sheet. Seismic refraction imaging is particularly well suited to delineating layering within the drumlin, and is able to reconstruct depths to interfaces to within ± 0.5 m accuracy. Refraction imaging ascertained that the weathered bedrock layer is continuous and of substantial thickness, so that it acts as a basal aquifer which underdrains the bulk of the drumlin. Electrical resistivity imaging was found to be capable of delineating relative spatial changes in the moisture content of the till units, as well as mapping sedimentary inclusions within the till. The moisture content appeared to be elevated near the margins of the drumlin, which may infer a weakening of the drumlin slopes. Our findings advocate the use of seismic refraction and electrical resistivity methods in future sedimentological and geotechnical studies of internal drumlin architecture and drumlin formation, owing particularly to the superior, 3- D spatial coverage of these methods.

Identification of gene networks associated with acute myeloid leukemia by comparative molecular methylation and expression profiling

Around 80% of acute myeloid leukemia (AML) patients achieve a complete remission, however many will relapse and ultimately die of their disease. The association between karyotype and prognosis has been studied extensively and identified patient cohorts as having favourable [e.g. t(8; 21), inv (16)/t(16; 16), t(15; 17)], intermediate [e.g. cytogenetically normal (NK-AML)] or adverse risk [e.g. complex karyotypes]. Previous studies have shown that gene expression profiling signatures can classify the sub-types of AML, although few reports have shown a similar feature by using methylation markers. The global methylation patterns in 19 diagnostic AML samples were investigated using the Methylated CpG Island Amplification Microarray (MCAM) method and CpG island microarrays containing 12,000 CpG sites. The first analysis, comparing favourable and intermediate cytogenetic risk groups, revealed significantly differentially methylated CpG sites (594 CpG islands) between the two subgroups. Mutations in the NPM1 gene occur at a high frequency (40%) within the NK-AML subgroup and are associated with a more favourable prognosis in these patients. A second analysis comparing the NPM1 mutant and wild-type research study subjects again identified distinct methylation profiles between these two subgroups. Network and pathway analysis revealed possible molecular mechanisms associated with the different risk and/or mutation sub-groups. This may result in a better classification of the risk groups, improved monitoring targets, or the identification of novel molecular therapies.

Electrical characterization of ALD Al2O3 and HfO2 films on germanium

Al2O3 and HfO2 films were deposited on germanium substrates by atomic layer deposition (ALD) and analyzed by MOS capacitor electrical characterization. In-situ plasma nitridation performed prior to ALD was found to improve the stability of the interface. For Al 2O3/GeON/Ge capacitors, a 450°C anneal in nitrogen ambient reduced hysteresis and oxide fixed charge to 90 mV and 1012 cm-2 respectively, with low leakage current density. On the contrary, degradation was observed for un-nitrided Al2O3/Ge capacitors after 300 and 400°C post-metal anneals. HfO2/GeON/Ge capacitors benefitted from a 400°C densification anneal but exhibited degradation after post-metal anneals at temperatures greater than 300°C. This degradation is attributed to the influence of Al electrodes on the HfO 2 gate stack. HfO2 is considered to be a suitable material for the gate stack and Al2O3 for the buried dielectric in a GeOI structure. ©The Electrochemical Society.

The inhibitor profiling of the caspase family of proteases using substrate-derived peptide glyoxals.

A series of substrate-based a-keto-ß-aldehyde (glyoxal) sequences have been synthesised and evaluated as inhibitors of the caspase family of cysteine proteases. A number of potent inhibitor sequences have been identified. For example, a palmitic acid containing sequence pal-Tyr-Val-Ala-Asp-glyoxal was demonstrated to be an extremely effective inhibitor of caspase-1, inhibiting not only the action of the protease against synthetic fluorogenic substrates (Ki = 0.3 nM) but also blocking its processing of pro-interleukin-1beta (pro-IL-1ß). In addition, the peptide Ac-Asp-Glu-Val-Asp-glyoxal, which is based on the consensus cleavage sequence for caspase-3, is a potent inhibitor of this protease (Ki = 0.26 nM) yet only functions as a comparatively modest inhibitor of caspase-1 (Ki = 451 nM). Potent inhibitor sequences were also identified for caspases-6 and -8. However, the degree of discrimination between the family members is limited. The ability of Ac-Asp-Glu-Val-Asp-glyoxal to block caspase-3 like activity in whole cells and to delay the development of apoptosis was assessed. When tested against caspase-3 like activity in cell lysates, Ac-Asp-Glu-Val-Asp-glyoxal displayed effective inhibition similar to that observed against recombinant caspase-3. Treatment of whole cells with this potent caspase-3 inhibitor was however, not sufficient to significantly stall the development of apoptosis in-vitro.

Plasma biomarker profiling in the detection of growth promoter use in calves

The detection of illicit growth promoter use during meat production within the European Union is reliant on residue testing which is a limiting factor on the number of animals which can be tested and consequently compromises the efficacy of testing procedures. The present study examined a novel detection strategy based on the profiling of plasma component concentrations in response to growth promoter administrations. Calves subjected to nortestosterone decanoate, 17 beta-oestradiol benzoate and dexamethasone were found to have altered urea, aminoterminal propeptide of type III procollagen and sex hormone binding globulin profiles in response to treatments. These findings demonstrate the potential of using the identification of perturbed profiles within a panel of biomarkers which cover a spectrum of biological activity to reveal growth promoter abuse.

Micro-Raman and Spreading Resistance Analysis on Beveled Implanted Germanium for Layer Transfer Applications

Raman and spreading resistance profiling have been used to analyze defects in germanium caused by hydrogen and helium implants, of typical fluences used in layer transfer applications. Beveling has been used to facilitate probing beyond the laser penetration depth. Results of Raman mapping along the projection area reveal that after post-implant annealing at 400°C, some crystal damage remains, while at 600°C, the crystal damage has been repaired. Helium implants create acceptor states beyond the projected range, and for both hydrogen and helium, 1×1016 acceptors/cm2 remain after 600°C. These are thought to be vacancy-related point defect clusters.

Cross-validation of geo-electrical and hydrogeological models to evaluate seawater intrusion in coastal aquifers

The validation of variable-density flow models simulating seawater intrusion in coastal aquifers requires information about concentration distribution in groundwater. Electrical resistivity tomography (ERT) provides relevant data for this purpose. However, inverse modeling is not accurate because of the non-uniqueness of solutions. Such difficulties in evaluating seawater intrusion can be overcome by coupling geophysical data and groundwater modeling. First, the resistivity distribution obtained by inverse geo-electrical modeling is established. Second, a 3-D variable-density flow hydrogeological model is developed. Third, using Archie's Law, the electrical resistivity model deduced from salt concentration is compared to the formerly interpreted electrical model. Finally, aside from that usual comparison-validation, the theoretical geophysical response of concentrations simulated with the groundwater model can be compared to field-measured resistivity data. This constitutes a cross-validation of both the inverse geo-electrical model and the groundwater model.
[Comte, J.-C., and O. Banton (2007), Cross-validation of geo-electrical and hydrogeological models to evaluate seawater intrusion in coastal aquifers, Geophys. Res. Lett., 34, L10402, doi:10.1029/2007GL029981.]