Modulation of agonist-stimulated second messenger and contractile events in bovine tracheal smooth muscle with cyclic nucleotide PDE inhibitors.

The aim of this thesis was to investigate and characterize the effects of isoenzyme- selective phosphodiesterase (PDE) inhibitors on a range of biochemical and functional parameters in bovine airway smooth muscle in order to further our understanding of 'cross-talk' between different second messenger pathways in this tissue.Cyclic nucleotide levels were measured after selective or non-selective PDE isoenzyme inhibition under conditions of basal and stimulated adenylyl and guanylyl cyclase activity. These data led to the conclusion that PDE IV plays an important role in the regulation of agonist-stimulated increases in cAMP levels, since under conditions of isoprenaline stimulation, incubation with the PDE IV-selective inhibitor rolipram resulted in an increase in cAMP from a control level of 20 2.7 to 50.3 4.0 pmol/mg protein. This response was potentiated in a synergistic manner by simultaneously inhibiting PDE III with ORG 9935. Co-inhibition of PDE III/IV also resulted in a significant increase in basal cAMP levels in tracheal smooth muscle slices and a marked decrease in the rolipram EC50 for cAMP accumulation in isoprenaline-stimulated slices from 205 102 to 7.3 3.0 muM. This suggested that metabolism of cAMP by PDE III is also functionally important in this tissue at least when PDE IV activity is compromized. Evidence was also obtained to suggest that cGMP is metabolized by PDE V, since incubation with zaprinast resulted in a 47% increase in basal cGMP values; however, where cGMP levels were elevated a greater increase in cGMP accumulation was seen in the presence of the nonspecific PDE inhibitor, IBMX (24.3 1.6 pmol/mg protein) compared to that seen with zaprinast (15.2 1.6 pmol/mg protein) suggesting that cGMP metabolism by other PDEs plays a significant role under these conditions. The effects of selective PDE inhibition on agonist-stimulated inositol phosphate accumulation was then investigated. Stimulation with maximally-effective concentrations of carbachol and histamine for 30 min resulted in 36- and 10-fold increases in inositol phosphate accumulations. The response to carbachol (1-100 muM) was largely unaffected by increases in cAMP accumulation caused by isoprenaline or PDE inhibition, however both manipulations inhibited the response to histamine (100muM) by approximately 80%. Again the inhibitory effects of rolipram were potentiated by ORG 9935 such that the EC50 value for rolipram-mediated inhibition was decreased from 120 27 to 1.5 0.9 muM. Such results suggest that PDE III/IV inhibition may be effective in relaxing airway smooth muscle. Consequently it was established that whilst rolipram could inhibit smooth muscle contraction stimulated by histamine or sub-maximal concentrations of methacholine, co-inhibition of PDEs with rolipram and ORG 9935 resulted in a much more potent anti-spasmogenic action. Furthermore, in contrast with the effects of rolipram alone the PDE III/IV inhibitor combination also significantly inhibited phasic contractions generated by either agonist. For example rolipram/ORG 9935 completely abolished the ability of 30uM histamine to cause a phasic contractile response. In common with previous suggestions that the membrane hyperpolarizing actions of cAMP-elevating agents may be responsible, at least in part, for the relaxation of trachealis muscle, experiments reported here suggest that PDE inhibition causes membrane hyperpolarization, possibly through increasing the open-state probability of the high- conductance, calcium-activated potassium channel, and this action may account for the mechanism whereby selective PDE inhibitors can inhibit phosphoinositide turnover and possibly as a consequence relax airway smooth muscle

Additional file 3: of Deep genome sequencing and variation analysis of 13 inbred mouse strains defines candidate phenotypic alleles, private variation and homozygous truncating mutations

Pairwise variant comparisons. The number of SNPs and indels shared between any two strains contained in the MGP variation catalogue. (XLSX 24 kb

Complete listing of the mutant mouse genes screened and the associated allele information

A subset of genes (29) were screened as 2 different alleles and for 1 gene a total of 3 alleles have been screened. The different alleles were generally tm1a and tm1b to discern if there was any difference after cre-mediated excision of the critical exon and targeting cassette (for details of the EUCOMM/KOMP-CSD tm1a/tm1b allele nomenclature refer to Skarnes et al 9). In some instances, one allele was a point mutation and the other targeting the whole gene. ‘Screened gene’ is the gene name as it appears in Data Record 2, followed by the ‘full allele name’, ‘allele superscript’ and ‘allele MGI ID’ (where a record exists). The ‘unique identifier’ is the internal colony prefix that was used for tracking and is specific to the gene and allele within the institute. In some instances, there are multiple colony prefixes for the same gene and allele – this occurred when additional mice were generated for testing outside of the high-throughput screening project. ‘MGI Gene/Marker ID’ allows tracking of the gene independent of any change in gene name for which a number have changed. The ‘current symbol’ is the most recent gene name and ‘synonym screened’ is where the gene has since been changed at MGI (and also internally) so allows for cross checking to the original data. The full current gene name and feature annotation is listed together with the chromosomal location of the gene and the Ensembl ID

Euphoria longana Lam.

原著和名: リウガン科名: ムクロジ科 = Sapindaceae採集地: 鹿児島県 肝属郡 佐多町 伊座敷 (大隅 肝属郡 佐多町 伊座敷)採集日: 1957/7/10採集者: 萩庭丈壽整理番号: JH026103国立科学博物館整理番号: TNS-VS-97610

Complete data set for all the mice comprising the 810 genes screened

Results of the B16-F10 pulmonary metastasis screen for the 810 genes consisting of 13,426 individual mice

Additional file 6: of Deep genome sequencing and variation analysis of 13 inbred mouse strains defines candidate phenotypic alleles, private variation and homozygous truncating mutations

Private SNPs with multiple VEP consequences. Private SNPs with multiple predicted VEP consequences for each strain strain. Consequence predictions are based on gene models obtained from Ensembl 78. (TXT 66669 kb

Additional file 1: Table S1. of Explorations to improve the completeness of exome sequencing

Coverage of exons in 54 probands. Table S2. Genes with low coverage exons in all 54 probands. (XLSX 9613 kb

Treatment response and neurofilament light chain levels with long-term patisiran in hereditary transthyretin-mediated amyloidosis with polyneuropathy: 24-month results of an open-label extension study

Longitudinal changes in neurofilament light chain (NfL) levels were evaluated alongside prespecified clinical assessments 24 months into the patisiran Global open-label extension (OLE) study in patients with ATTRv amyloidosis with polyneuropathy. All patients enrolled in the Global OLE, from phase III APOLLO and phase II OLE parent studies, received patisiran. Assessments included measures of polyneuropathy (modified Neuropathy Impairment Score+7 (mNIS+7)), quality of life (QOL; Norfolk QOL-Diabetic Neuropathy questionnaire (Norfolk QOL-DN)), and plasma NfL. Patients receiving patisiran in the parent study (APOLLO-patisiran, n = 137; phase II OLE-patisiran, n = 25) demonstrated sustained improvements in mNIS+7 (mean change from parent study baseline (95% confidence interval): APOLLO-patisiran −4.8 (−8.9, −0.6); phase II OLE-patisiran −5.8 (−10.5, −1.2)) and Norfolk QOL-DN (APOLLO-patisiran −2.4 (−7.2, 2.3)), and maintained reduced NfL levels at Global OLE 24 months. After initiating patisiran in the Global OLE, APOLLO-placebo patients (n = 49) demonstrated stabilized mNIS+7, improved Norfolk QOL-DN, and significantly reduced NfL levels. Patisiran continued to demonstrate an acceptable safety profile. Earlier patisiran initiation was associated with a lower exposure-adjusted mortality rate. Long-term patisiran treatment led to sustained improvements in neuropathy and QOL, with NfL demonstrating potential as a biomarker for disease progression and treatment response in ATTRv amyloidosis with polyneuropathy.</p

MOESM1 of Retrospective analysis in oculocutaneous albinism patients for the 2.7Â kb deletion in the OCA2 gene revealed a co-segregation of the controversial variant, p.R305W

Additional file 1: Table S1. Primers for TYR intronic regions

Additional file 3: Figure S2. of Somatic drivers of B-ALL in a model of ETV6-RUNX1; Pax5 +/− leukemia

Accelerated tumor development in Ink4a−/−; Etv6 +/RUNX1-SB mice. (PPT 162 kb