Uptake, biodistribution, and time course of naked plasmid DNA trafficking after intratumoral in vivo jet injection

Nonviral jet injection is an applicable technology for in vivo gene transfer of naked DNA. However, little is known about the biodistribution and clearance of jet-injected DNA, or about its localization within tissue and cells. Therefore, in this study we analyzed the intratumoral and systemic biodistribution of jet-injected naked DNA in human colon carcinoma-bearing NCr-nu/nu mice, which were jet-injected with the pCMVbeta plasmid DNA. Intratumoral and systemic plasmid DNA biodistribution was analyzed 5, 10, 20, and 40 min and 3, 6, 24, 48, and 72 hr after jet injection, using quantitative real-time polymerase chain reaction. In the tumors, a rapid drop in naked DNA load within 24 hr of jet injection was shown. Detailed analysis of intratumoral distribution of rhodamine-labeled DNA revealed the presence of plasmid DNA within tumor cells 5 min after jet injection and further accumulation of significant DNA amounts in the cell nuclei 30 to 60 min after jet injection. In the blood, DNA amounts rapidly dropped within 10 to 40 min of jet injection to less than 0.001 pg of plasmid per 250 ng of tissue DNA and only minimal plasmid DNA dissemination was detected in liver, lung, spleen, kidney, and ovaries, which was cleared 3 to 6 hr after jet injection. By contrast, in heart, bone marrow, and brain almost no plasmid DNA was detectable

Flexible provisioning of Web service workflows

Web services promise to revolutionise the way computational resources and business processes are offered and invoked in open, distributed systems, such as the Internet. These services are described using machine-readable meta-data, which enables consumer applications to automatically discover and provision suitable services for their workflows at run-time. However, current approaches have typically assumed service descriptions are accurate and deterministic, and so have neglected to account for the fact that services in these open systems are inherently unreliable and uncertain. Specifically, network failures, software bugs and competition for services may regularly lead to execution delays or even service failures. To address this problem, the process of provisioning services needs to be performed in a more flexible manner than has so far been considered, in order to proactively deal with failures and to recover workflows that have partially failed. To this end, we devise and present a heuristic strategy that varies the provisioning of services according to their predicted performance. Using simulation, we then benchmark our algorithm and show that it leads to a 700% improvement in average utility, while successfully completing up to eight times as many workflows as approaches that do not consider service failures

Evidence of Spawning Capable Tarpon (Megalops atlanticus) Off the Louisiana Coast

Despite the fact that the tarpon (Megalops atlanticus) is a popular sport fish in the Gulf of Mexico (GOM; Ault et al. 2008), little is known of its spawning behavior. Spawning M. atlanticus have never been documented and fertilized eggs have not been observed in situ (Ault et al. 2008). While it has been suggested that adult M. atlanticus move to deep water off the southwest coast of Florida and into the Yucatan Channel to spawn, the actual locations of spawning grounds remain unknown (Smith 1980, Crabtree et al. 1995, Crabtree et al. 1997). Distribution patterns of larvae (leptocephali) have served as the basis for most of what has been inferred about the spawning areas (Smith 1980, Crabtree et al. 1997). For example, leptocephali were collected from the southwestern GOM, the Yucatan Channel, and along the west coast of Florida, and based on the their size, it was assumed that M. atlanticus spawned in nearby areas (Smith 1980). Histological examination of gonads has also been used to estimate the location of M. atlanticus spawning habitat. Females collected from the Florida Keys and Boca Grande Pass off the west coast of Florida and contained ovaries with post ovulatory follicles (POF) and advanced vitellogenic oocytes, suggesting M. atlanticus spawn in this region from April through July (Crabtree et al. 1997). Examination of gonads from M. atlanticus caught off the coast of equatorial Ceara State, Brazil suggested that spawning occurs there from October through January (de Menezes and Paiva 1966). We report here the first evidence of spawning capable M. atlanticus off the coast of Louisiana in the northern GOM based on histological examination of gonads

EMPOWERing older people and their communities to manage their own CARE (EMPOWERCARE): Evaluation study of a social innovation initiative across four European countries

This paper outlines the evaluation strategy of the EMPOWERCARE, an EU Interreg 2 Seas funded social innovation project, which involves a partnership of local authorities, universities and non-governmental organisations from four European countries: Belgium, France, the Netherlands and the United Kingdom. Partners are working in collaboration with local people aged 65+, and those aged 50+ with at least one chronic condition, to respond to current gaps in the care of older people. The project aims specifically to contribute to person-centred care and technology knowledge transfer whilst reducing isolation, loneliness and increasing solidarity amongst older people and their communities. During the project, partners are jointly developing a strategy for implementing good-practice models, a workforce transformation approach via shared training and a technology blueprint with emphasis on caring digital technologies to address the rising demand for health and social care services for older people. The project addresses this challenge by drawing on community assets, involving older people in decisions about their own health and wellbeing, enabling them to keep healthier and in their own homes and communities safely for longer. The project’s evaluation aims to measure the impact of the EMPOWERCARE initiative across 7 pilot sites in the 4 participating countries. Given the Covid-19 pandemic, the evaluation design has been developed with a view to being agile and able to respond to complex and shifting situations, and especially so in terms of what and how data are collected. A realist synthesis approach (Pawson & Tilley, 2004) guides the evaluation within a descriptive case study design (Yin 2003) to identify and contextualise the project strategies that are influential within and across the 7 different case study sites. The evaluation uses multiple interdisciplinary methods, such as surveys, Photovoice and Social Return on Investment, to capture a range of perspectives across three timepoints, baseline (T0), mid-point (T1) and end-point (T2). Online surveys are conducted with both end-users and the workforce in all pilot sites at T0 and T2. At T1, end-users are actively involved in creating visual data through Photovoice to capture their lived experiences with local initiatives in pilot sites. Visual data are going to be displayed at pilot site exhibitions targeted to key local stakeholders and members of the workforce. Focus groups will be conducted with all participants reflecting on the visual data exhibition to explore insights about the initiative from current and future perspectives. This aspect is informed by a Social Return on Investment approach. The sequential data collection of multiple sources and longitudinal study design identify patterns of change and impact. The evaluation design will deliver a theory-driven rich explanation of what works about EMPOWERCARE and for whom, why, how and in what circumstances it works. The analysis of the evaluation findings will contribute to a more empowered person-centred approach, with more inclusive and caring digital health solutions that will allow policy makers to develop localised, efficient and social-value driven services to meet the rising needs of older people in the European regions

Renormalon Model Predictions for Power-Corrections to Flavour Singlet Deep Inelastic Structure Functions

We analyze power corrections to flavour singlet deep inelastic scattering structure functions in the framework of the infrared renormalon model. Our calculations, together with previous results for the non-singlet contribution, allow to model the x-dependence of higher twist corrections to F_2, F_L and g_1 in the whole x domain.Comment: LaTeX, 25 pages, 8 eps figures included, one figure was added. Final version for publication in Nucl.Phys.

Genetic deletion of hepatic NCOR1 protects from atherosclerosis by promoting alternative bile acid-metabolism and sterol excretion

BACKGROUND The nuclear receptor corepressor 1 (NCOR1) plays an important role in the regulation of gene expression in immunometabolic conditions by connecting chromatin-modifying enzymes, coregulators and transcription factors. NCOR1 has been shown to be involved in cardiometabolic diseases. Recently, we demonstrated that the deletion of macrophage NCOR1 aggravates atherosclerosis by promoting CD36-triggered foam cell formation via PPARG derepression. PURPOSE Since NCOR1 modulates the function of several key regulators involved in hepatic lipid and bile acid metabolism, we hypothesized that its deletion in hepatocytes alters lipid metabolism and atherogenesis. METHODS To test this hypothesis, we generated hepatocyte-specific Ncor1 knockout mice on a Ldlr-/- background. Besides assessing the progression of the disease in thoracoabdominal aortae en face, we analyzed hepatic cholesterol and bile acid metabolism at expression and functional levels. RESULTS Our data demonstrate that liver-specific Ncor1 knockout mice on an atherosclerosis-prone background develop less atherosclerotic lesions than controls. Interestingly, under chow diet, plasma cholesterol levels of liver-specific Ncor1 knockout mice were slightly higher compared to control, but strongly reduced compared to control mice after feeding them an atherogenic diet for 12 weeks. Moreover, the hepatic cholesterol content was decreased in liver-specific Ncor1 knockout compared to control mice. Our mechanistic data revealed that NCOR1 reprograms the synthesis of bile acids towards the alternative pathway, which in turn reduce bile hydrophobicity and enhances fecal cholesterol excretion. CONCLUSIONS Our data suggest that hepatic Ncor1 deletion in mice decreases atherosclerosis development by reprograming bile acid metabolism and enhancing fecal cholesterol excretion

A standard variation file format for human genome sequences

Here we describe the Genome Variation Format (GVF) and the 10Gen dataset. GVF, an extension of Generic Feature Format version 3 (GFF3), is a simple tab-delimited format for DNA variant files, which uses Sequence Ontology to describe genome variation data. The 10Gen dataset, ten human genomes in GVF format, is freely available for community analysis from the Sequence Ontology website and from an Amazon elastic block storage (EBS) snapshot for use in Amazon's EC2 cloud computing environment

Feasibility of a hyper-acute stroke unit model of care across England: a modelling analysis.

The policy of centralising hyperacute stroke units (HASUs) in England aims to provide stroke care in units that are both large enough to sustain expertise (>600 admissions/year) and dispersed enough to rapidly deliver time-critical treatments (<30 min maximum travel time). Currently, just over half (56%) of patients with stroke access care in such a unit. We sought to model national configurations of HASUs that would optimise both institutional size and geographical access to stroke care, to maximise the population benefit from the centralisation of stroke care.This article is freely available via Open Access. Click on the Additional Link above to access the full-text via the publisher's site.Publishe

An Analysis of the Next-to-Leading Order Corrections to the g_T(=g_1+g_2) Scaling Function

We present a general method for obtaining the quantum chromodynamical radiative corrections to the higher-twist (power-suppressed) contributions to inclusive deep-inelastic scattering in terms of light-cone correlation functions of the fundamental fields of quantum chromodynamics. Using this procedure, we calculate the previously unknown ${\cal O}(\alpha_s)$ corrections to the twist-three part of the spin scaling function $g_T(x_B,Q^2) (=g_1(x_B,Q^2)+g_2(x_B,Q^2))$ and the corresponding forward Compton amplitude $S_T(\nu,Q^2)$. Expanding our result about the unphysical point $x_B=\infty$, we arrive at an operator product expansion of the nonlocal product of two electromagnetic current operators involving twist-two and -three operators valid to ${\cal O}(\alpha_s)$ for forward matrix elements. We find that the Wandzura-Wilczek relation between $g_1(x_B,Q^2)$ and the twist-two part of $g_T(x_B,Q^2)$ is respected in both the singlet and non-singlet sectors at this order, and argue its validity to all orders. The large-$N_c$ limit does not appreciably simplify the twist-three Wilson coefficients.Comment: 41 pages, 9 figures, corrected minor erro

Nematode.net update 2008: improvements enabling more efficient data mining and comparative nematode genomics

Nematode.net (http://nematode.net) is a publicly available resource dedicated to the study of parasitic nematodes. In 2000, the Genome Center at Washington University (GC) joined a consortium including the Nematode Genomics group in Edinburgh, and the Pathogen Sequencing Unit of the Sanger Institute to generate expressed sequence tags (ESTs) as an inexpensive and efficient solution for gene discovery in parasitic nematodes. As of 2008 the GC, sampling key parasites of humans, animals and plants, has generated over 500 000 ESTs and 1.2 million genome survey sequences from more than 30 non-Caenorhabditis elegans nematodes. Nematode.net was implemented to offer user-friendly access to data produced by this project. In addition to sequence data, the site hosts: assembled NemaGene clusters in GBrowse views characterizing composition and protein homology, functional Gene Ontology annotations presented via the AmiGO browser, KEGG-based graphical display of NemaGene clusters mapped to metabolic pathways, codon usage tables, NemFam protein families which represent conserved nematode-restricted coding sequences not found in public protein databases, a web-based WU-BLAST search tool that allows complex querying and other assorted resources. The primary aim of Nematode.net is the dissemination of this diverse collection of information to the broader scientific community in a way that is useful, consistent, centralized and enduring