3,305 research outputs found
Targeted next-generation sequencing for routine clinical screening of mutations
In many fields it is now desirable to sequence large panels of genes for mutation, to aid management of patients. The need for extensive sample preparation means that current approaches for assessing mutation status in the clinical setting are limited. A recent publication demonstrates a single-step, targeted, true single-molecule sequencing approach to assessing the mutational status of BRCA1. Fragmented DNA samples are loaded directly onto a flow cell and sequenced, thus detecting both small- and large-scale mutations with minimal sample preparation and high accuracy
Cost minimisation study of transdermal glyceryl trinitrate in reducing failures of peripheral intravenous infusion
This article has been made available through the Brunel Open Access Publishing Fund and is available from the specified link - Copyright @ 1989 BMJ Publishing Group
High affinity binding of the peptide agonist TIP-39 to the parathyroid hormone 2 (PTHâ) receptor requires the hydroxyl group of Tyr-318 on transmembrane helix 5
TIP39 (âtuberoinfundibular peptide of 39 residuesâ) acts via the parathyroid hormone 2 receptor, PTHâ, a Family B G protein-coupled receptor (GPCR). Despite the importance of GPCRs in human physiology and pharmacotherapy, little is known about the molecular details of the TIP39-PTHâ interaction. To address this, we utilised the different pharmacological profiles of TIP39 and PTH(1-34) at PTHâ and its related receptor PTHâ: TIP39 being an agonist at the former but an antagonist at the latter, while PTH(1-34) activates both. A total of 23 site-directed mutations of PTHâ, in which residues were substituted to the equivalent in PTHâ, were made and pharmacologically screened for agonist activity. Follow-up mutations were analysed by radioligand binding and cAMP assays. A model of the TIP39-PTHâ complex was built and analysed using molecular dynamics. Only Tyr318-Ile displayed reduced TIP39 potency, despite having increased PTH(1-34) potency, and further mutagenesis and analysis at this site demonstrated that this was due to reduced TIP39 affinity at Tyr318-Ile (pIC50 = 6.01±0.03) compared with wild type (pICâ
â = 7.81±0.03). The hydroxyl group of the Tyr-318âs side chain was shown to be important for TIP39 binding, with the Tyr318-Phe mutant displaying 13-fold lower affinity and 35-fold lower potency compared with wild type. TIP39 truncated by up to 5 residues at the N-terminus was still sensitive to the mutations at Tyr-318, suggesting that it interacts with a region within TIP39(6-39). Molecular modelling and molecular dynamics simulations suggest that the selectivity is based on an interaction between the Tyr-318 hydroxyl group with the carboxylate side chain of Asp-7 of the peptide
"L'art poétique à l'usage du Petit Séminaire de Québec"
Y-a-t-il un « art poĂ©tique » surrĂ©aliste ? Si l'on en croit AndrĂ© Breton, non : lui se situe rĂ©solument du cĂŽtĂ© du « Manifeste », oĂč l'art poĂ©tique se voit dĂ©niĂ© sur ses deux versants - celui de l'artefact (techniques rhĂ©toriques) et celui du poĂ©tique, engagĂ© dans une « littĂ©rature » marquĂ©e ici du sceau de l'infamie. Le rationalisme Ă©clate sous la pression d'une « raison Ă©largie ». SimultanĂ©ment, l'esprit du manifeste fait triompher les voies du dĂ©sir : le « il y aura une fois » de « l'imagination ». D'oĂč une tĂ©lĂ©ologie de l'art, orientĂ© vers sa « prophĂ©tie », et une Ă©pistĂ©mologie de sa rĂ©ception, fondĂ©e sur l'Ă©ros. Ce qui suppose de ne pas glisser aux « techniques » qui mĂšnent aux poncifs : d'oĂč la Terreur.Is there a surrealist "poetic art" ? If we are to believe AndrĂ© Breton, the answer is No ; Breton for his part resolutely adheres to the "Manifesto". The possibility of a poetic art is denied both as art (rhetorical technique) and as poetry (a literary project here under the seal of infamy. The rationale of setting poetics in order collapses under the constraint of a "broader reason". At the same time, the spirit of the "Manifesto" marks the triumph of the ways of dĂ©sire: the imperative "let there be" of the imagination. Hence there is a teleology of Art founded on "prophecy", and an epistemology of its rĂ©ception based on Eros.The besetting danger is that "technique" (and consequently clichĂ©) will predominate : hence the Terror
Mobile element insertions are frequent in oesophageal adenocarcinomas and can mislead paired-end sequencing analysis.
BACKGROUND: Mobile elements are active in the human genome, both in the germline and cancers, where they can mutate driver genes. RESULTS: While analysing whole genome paired-end sequencing of oesophageal adenocarcinomas to find genomic rearrangements, we identified three ways in which new mobile element insertions appear in the data, resembling translocation or insertion junctions: inserts where unique sequence has been transduced by an L1 (Long interspersed element 1) mobile element; novel inserts that are confidently, but often incorrectly, mapped by alignment software to L1s or polyA tracts in the reference sequence; and a combination of these two ways, where different sequences within one insert are mapped to different loci. We identified nine unique sequences that were transduced by neighbouring L1s, both L1s in the reference genome and L1s not present in the reference. Many of the resulting inserts were small fragments that include little or no recognisable mobile element sequence. We found 6 loci in the reference genome to which sequence reads from inserts were frequently mapped, probably erroneously, by alignment software: these were either L1 sequence or particularly long polyA runs. Inserts identified from such apparent rearrangement junctions averaged 16 inserts/tumour, range 0-153 insertions in 43 tumours. However, many inserts would not be detected by mapping the sequences to the reference genome, because they do not include sufficient mappable sequence. To estimate total somatic inserts we searched for polyA sequences that were not present in the matched normal or other normals from the same tumour batch, and were not associated with known polymorphisms. Samples of these candidate inserts were verified by sequencing across them or manual inspection of surrounding reads: at least 85 % were somatic and resembled L1-mediated events, most including L1Hs sequence. Approximately 100 such inserts were detected per tumour on average (range zero to approximately 700). CONCLUSIONS: Somatic mobile elements insertions are abundant in these tumours, with over 75 % of cases having a number of novel inserts detected. The inserts create a variety of problems for the interpretation of paired-end sequencing data.Funding
was primarily from Cancer Research UK program grants to RCF and ST
(C14478/A15874 and C14303/A17197), with additional support awarded to
RCF from UK Medical Research Council, NHS National Institute for Health
Research (NIHR), the Experimental Cancer Medicine Centre Network and
the NIHR Cambridge Biomedical Research Centre, and Cancer Research UK
Project grant C1023/A14545 to PAWE. JMJW was funded by a Wellcome
Trust Translational Medicine and Therapeutics grant
Testing the translational power of the zebrafish: An interspecies analysis of responses to cardiovascular drugs.
This is the final version. Available from Frontiers Media via the DOI in this record.âŻDATA AVAILABILITY:
All datasets generated for this study are included in the
manuscript and the supplementary files.The zebrafish is rapidly emerging as a promising alternative in vivo model for the detection of drug-induced cardiovascular effects. Despite its increasing popularity, the ability of this model to inform the drug development process is often limited by the uncertainties around the quantitative relevance of zebrafish responses compared with nonclinical mammalian species and ultimately humans. In this test of concept study, we provide a comparative quantitative analysis of the in vivo cardiovascular responses of zebrafish, rat, dog, and human to three model compounds (propranolol, losartan, and captopril), which act as modulators of two key systems (beta-adrenergic and renin-angiotensin systems) involved in the regulation of cardiovascular functions. We used in vivo imaging techniques to generate novel experimental data of drug-mediated cardiovascular effects in zebrafish larvae. These data were combined with a database of interspecies mammalian responses (i.e., heart rate, blood flow, vessel diameter, and stroke volume) extracted from the literature to perform a meta-analysis of effect size and direction across multiple species. In spite of the high heterogeneity of study design parameters, our analysis highlighted that zebrafish and human responses were largely comparable in >80% of drug/endpoint combinations. However, it also revealed a high intraspecies variability, which, in some cases, prevented a conclusive interpretation of the drug-induced effect. Despite the shortcomings of our study, the meta-analysis approach, combined with a suitable data visualization strategy, enabled us to observe patterns of response that would likely remain undetected with more traditional methods of qualitative comparative analysis. We propose that expanding this approach to larger datasets encompassing multiple drugs and modes of action would enable a rigorous and systematic assessment of the applicability domain of the zebrafish from both a mechanistic and phenotypic standpoint. This will increase the confidence in its application for the early detection of adverse drug reactions in any major organ system.Biotechnology and Biological Sciences Research CouncilAstraZenec
Variation in the Large-Scale Organization of Gene Expression Levels in the Hippocampus Relates to Stable Epigenetic Variability in Behavior
Despite sharing the same genes, identical twins demonstrate substantial variability in behavioral traits and in their risk for disease. Epigenetic factors-DNA and chromatin modifications that affect levels of gene expression without affecting the DNA sequence-are thought to be important in establishing this variability. Epigenetically-mediated differences in the levels of gene expression that are associated with individual variability traditionally are thought to occur only in a gene-specific manner. We challenge this idea by exploring the large-scale organizational patterns of gene expression in an epigenetic model of behavioral variability.To study the effects of epigenetic influences on behavioral variability, we examine gene expression in genetically identical mice. Using a novel approach to microarray analysis, we show that variability in the large-scale organization of gene expression levels, rather than differences in the expression levels of specific genes, is associated with individual differences in behavior. Specifically, increased activity in the open field is associated with increased variance of log-transformed measures of gene expression in the hippocampus, a brain region involved in open field activity. Early life experience that increases adult activity in the open field also similarly modifies the variance of gene expression levels. The same association of the variance of gene expression levels with behavioral variability is found with levels of gene expression in the hippocampus of genetically heterogeneous outbred populations of mice, suggesting that variation in the large-scale organization of gene expression levels may also be relevant to phenotypic differences in outbred populations such as humans. We find that the increased variance in gene expression levels is attributable to an increasing separation of several large, log-normally distributed families of gene expression levels. We also show that the presence of these multiple log-normal distributions of gene expression levels is a universal characteristic of gene expression in eurkaryotes. We use data from the MicroArray Quality Control Project (MAQC) to demonstrate that our method is robust and that it reliably detects biological differences in the large-scale organization of gene expression levels.Our results contrast with the traditional belief that epigenetic effects on gene expression occur only at the level of specific genes and suggest instead that the large-scale organization of gene expression levels provides important insights into the relationship of gene expression with behavioral variability. Understanding the epigenetic, genetic, and environmental factors that regulate the large-scale organization of gene expression levels, and how changes in this large-scale organization influences brain development and behavior will be a major future challenge in the field of behavioral genomics
Carbamazepine-Induced Tics
A variety of movement disorders are known to occur in association with carbamazepine (CBZ) therapy in adults and children, but development of tics has been described infrequently and only in patients with underlying Tourette's syndrome or other movement disorders. We report 3 children with epilepsy who developed facial motor tics after initiation of CBZ for complex partial seizures. All 3 had documented CBZ blood levels in the therapeutic range at the time, and none had other symptoms or signs of clinical intoxication. Neurologic examinations were normal in 2 and showed developmental de lay of expressive language in the third. Brain imaging was normal in all. After development of the tics in 2, CBZ was continued at the same or higher dose, and the tics abated and then ceased spontaneously â€6 months. In the third child, the tics ceased after CBZ discontinuation. These cases demonstrate that CBZ can induce simple motor tics in children. These idiosyncratic reactions may be transient and do not always necessitate drug discontinuation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66313/1/j.1528-1157.1993.tb02119.x.pd
ESHAP and G-CSF is a superior blood stem cell mobilizing regimen compared to cyclophosphamide 1.5 g mâ2 and G-CSF for pre-treated lymphoma patients: a matched pairs analysis of 78 patients
Cyclophosphamide 1.5 g mâ2followed by granulocyte colony-stimulating factor (G-CSF) is an effective peripheral blood stem cell (PBSC) mobilizing regimen, but has limited anti-lymphoma activity. We therefore assessed the mobilizing potential of ESHAP (etoposide, ara-C, methylprednisolone and cisplatin), a potent second-line lymphoma regimen followed by G-CSF. The results were compared in 78 patients with relapsed or resistant lymphomas with the use of cyclophosphamide 1.5 g mâ2followed by G-CSF in a matched pairs analysis, matching the ESHAP recipients (for predetermined prognostic factors) from a cohort of 178 lymphoma patients mobilized with cyclophosphamide and G-CSF. The total numbers of mononuclear cells collected at apheresis was similar with both regimens but ESHAP plus G-CSF resulted in a significantly higher percentage of CD34+ cells, absolute number of CD34+ cells and GM-CFC (all with P -values < 0.001). The number of patients requiring only one apheresis harvest to achieve a CD34+ cell yield of > 2.0 Ă 106kgâ1was greatly increased in the ESHAP recipients (56/78 vs 17/78, P< 0.001). The total number of progenitor cells collected was not significantly different with the two mobilization regimens because of this higher number of apheresis in the cyclophosphamide group. The proportion of patients who failed to achieve a minimum CD34+ cell target of 1 Ă 106kgâ1with the pooled harvests was less in the ESHAP arm (four patients vs nine patients) despite an increased number of aphereses in the cyclophosphamide recipients. ESHAP plus G-CSF is well tolerated and is an excellent mobilization regimen in patients with pre treated lymphoma. © 2000 Cancer Research Campaig
Fluctuation scaling in complex systems: Taylor's law and beyond
Complex systems consist of many interacting elements which participate in
some dynamical process. The activity of various elements is often different and
the fluctuation in the activity of an element grows monotonically with the
average activity. This relationship is often of the form "", where the exponent is predominantly in
the range . This power law has been observed in a very wide range of
disciplines, ranging from population dynamics through the Internet to the stock
market and it is often treated under the names \emph{Taylor's law} or
\emph{fluctuation scaling}. This review attempts to show how general the above
scaling relationship is by surveying the literature, as well as by reporting
some new empirical data and model calculations. We also show some basic
principles that can underlie the generality of the phenomenon. This is followed
by a mean-field framework based on sums of random variables. In this context
the emergence of fluctuation scaling is equivalent to some corresponding limit
theorems. In certain physical systems fluctuation scaling can be related to
finite size scaling.Comment: 33 pages, 20 figures, 2 tables, submitted to Advances in Physic
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