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AimConstitutive release of NO blunts intrinsic and stimulated contractile activity in cerebral arteries (CA). Here, we explored whether phosphorylation and expression levels of the PKG-sensitive, leucine zipper positive (LZ(+)) splice variants of the regulatory subunit of myosin phosphatase (MYPT1) are involved and whether its expression is associated with higher cGMP sensitivity. MethodsVascular contractility was investigated by wire myography. Phosphorylation of MYPT1 was determined by Western blotting. ResultsConstitutive phosphorylation of MYPT1-T696 and T853 was lower and that of S695 and S668 was higher in cerebral arteries from the circulus arteriosus (CA-w) than in femoral arteries (FA), while total MYPT1 expression was not different. In CA-w but not in FA, L-NAME lowered phosphorylation of S695/S668 and increased phosphorylation of T696/T853 and of MLC20-S19, plus basal tone. The increase in basal tone was attenuated in CA-w and basilar arteries (BA) from heterozygous MYPT1-T696A/+ mice. Compared to FA, expression of the LZ(+)-isoform was 2-fold higher in CA-w coincident with a higher sensitivity to DEA-NONOate, cinaciguat and Y27632 in BA and 8-Br-cGMP (1 mol/L) in pre-constricted (pCa 6.1) -toxin permeabilized CAs. In contrast, 6-Bnz-cAMP (10 mol/L) relaxed BA and FA similarly by 80%. ConclusionOur results indicate that (i) regulation of the intrinsic contractile activity in CA involves phosphorylation of MYPT1 at T696 and S695/S668, (ii) the higher NO/cGMP/PKG sensitivity of CAs can be ascribed to the higher expression level of the LZ(+)-MYPT1 isoform and (iii) relaxation by cAMP/PKA pathway is less dependent on the expression level of the LZ(+) splice variants of MYPT1.

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Ore was processed from 1956 to 1984 at Moab, Utah, USA. Interim cover placement on the tailings pile was completed in 1995 and in 1990 pumping of the water began from wells in the pile to the top of the pile to accelerate dewatering and consolidation. In 2000 a system of vertical band drains or wicks was installed, which penetrated the centre of the tailings pile in a triangular pattern. The wicks were connected to a manifold which allowed gravity drainage to a sump, and a small evaporation pond was constructed for the elimination of collected water. Excavation of the pile in 2010 cut into the wicks, several of which began to discharge pore water with a pH as low as 2.5 and high concentrations of ammonia, U and salts. A project was developed to safely handle the pore water while allowing excavation and conditioning activities to continue. Earthen berms of fine-grained tailings were constructed within the contaminated area near the base of the excavation to contain the pore water. Seven ponds were constructed capable of holding 2 400 000 l while preparations were made to eliminate the water. Groundwater contaminated with ammonia and U is pumped to a 1.6 ha lined pond on top of the tailings pile. The pond promotes evaporation to eliminate the water and acts as temporary storage until trucks withdraw the water and apply it on the tailings pile and roads in the contaminated areas for dust control. By October 2010 the amount of pore water was substantially reduced and most of the ponds were eliminated or consolidated into two ponds containing less than 380 000 l, and little additional pore water was encountered as at June 2011., Ore was processed from 1956 to 1984 at Moab, Utah, USA. Interim cover placement on the tailings pile was completed in 1995 and in 1990 pumping of the water began from wells in the pile to the top of the pile to accelerate dewatering and consolidation. In 2000 a system of vertical band drains or wicks was installed, which penetrated the centre of the tailings pile in a triangular pattern. The wicks were connected to a manifold which allowed gravity drainage to a sump, and a small evaporation pond was constructed for the elimination of collected water. Excavation of the pile in 2010 cut into the wicks, several of which began to discharge pore water with a pH as low as 2.5 and high concentrations of ammonia, U and salts. A project was developed to safely handle the pore water while allowing excavation and conditioning activities to continue. Earthen berms of fine-grained tailings were constructed within the contaminated area near the base of the excavation to contain the pore water. Seven ponds were constructed capable of holding 2 400 000 l while preparations were made to eliminate the water. Groundwater contaminated with ammonia and U is pumped to a 1.6 ha lined pond on top of the tailings pile. The pond promotes evaporation to eliminate the water and acts as temporary storage until trucks withdraw the water and apply it on the tailings pile and roads in the contaminated areas for dust control. By October 2010 the amount of pore water was substantially reduced and most of the ponds were eliminated or consolidated into two ponds containing less than 380 000 l, and little additional pore water was encountered as at June 2011.

Anaplastic large cell lymphoma (ALCL) is a main type of T-cell lymphomas and comprises three distinct entities: systemic anaplastic lymphoma kinase (ALK) positive, systemic ALK(-) and cutaneous ALK(-) ALCL (cALCL). Little is known about their pathogenesis and their cellular origin, and morphological and immunophenotypical overlap exists between ALK(-) ALCL and classical Hodgkin lymphoma (cHL). We conducted gene expression profiling of microdissected lymphoma cells of five ALK(+) and four ALK(-) systemic ALCL, seven cALCL and sixteen cHL, and of eight subsets of normal T and NK cells. The analysis supports a derivation of ALCL from activated T cells, but the lymphoma cells acquired a gene expression pattern hampering an assignment to a CD4(+), CD8(+) or CD30(+) T-cell origin. Indeed, ALCL display a down-modulation of many T-cell characteristic molecules. All ALCL types show significant expression of NFkappaB target genes and upregulation of genes involved in oncogenesis (e.g. EZH2). Surprisingly, few genes are differentially expressed between systemic and cALCL despite their different clinical behaviour, and between ALK(-) ALCL and cHL despite their different cellular origin. ALK(+) ALCL are characterized by expression of genes regulated by pathways constitutively activated by ALK. This study provides multiple novel insights into the molecular biology and pathogenesis of ALCL.

BACKGROUND: Although current molecular clock methods offer greater flexibility in modelling evolutionary events, calibration of the clock with dates from the fossil record is still problematic for many groups. Here we implement several new approaches in molecular dating to estimate the evolutionary ages of Lacertidae, an Old World family of lizards with a poor fossil record and uncertain phylogeny. Four different models of rate variation are tested in a new program for Bayesian phylogenetic analysis called TreeTime, based on a combination of mitochondrial and nuclear gene sequences. We incorporate paleontological uncertainty into divergence estimates by expressing multiple calibration dates as a range of probabilistic distributions. We also test the reliability of our proposed calibrations by exploring effects of individual priors on posterior estimates. RESULTS: According to the most reliable model, as indicated by Bayes factor comparison, modern lacertids arose shortly after the K/T transition and entered Africa about 45 million years ago, with the majority of their African radiation occurring in the Eocene and Oligocene. Our findings indicate much earlier origins for these clades than previously reported, and we discuss our results in light of paleogeographic trends during the Cenozoic. CONCLUSION: This study represents the first attempt to estimate evolutionary ages of a specific group of reptiles exhibiting uncertain phylogenetic relationships, molecular rate variation and a poor fossil record. Our results emphasize the sensitivity of molecular divergence dates to fossil calibrations, and support the use of combined molecular data sets and multiple, well-spaced dates from the fossil record as minimum node constraints. The bioinformatics program used here, TreeTime, is publicly available, and we recommend its use for molecular dating of taxa faced with similar challenges.