Vision and Valuation of a Citizen-Centric Shared Information Portal

The administrative burden the government puts on citizens is substantial, whereas, generally speaking, service levels are low and a ‘customer’ orientation is lacking. There is a growing understanding that e-government can play an important role in tackling these issues by better exchange of information and electronic availability. This paper reports on the development and evaluation of an e-government vision as part of a strategic planning trajectory for the social security sector and other government agencies in the Netherlands. The vision approaches governmental service delivery from the citizen viewpoint and helps governmental organisations to take service- and citizen orientation to a higher level. The concepts used in the vision were tested by boardroom sessions as well as a survey, and has become the guiding principles for a number of e-government developments

Implementing Electronic Services Transnational Guidelines and Perspectives

Electronic services to citizens are a growing concern to governments all over the world, not inthe least in the domains of social security and labor market. It was at the Montreal Conference of the ISSA –the International Social Security Association – in 1999 that many organizations in many countries showedto be grappling with many questions concerning the implementation of electronic service delivery. In orderto elaborate on experiences of implementation, the ISSA and three Dutch member organizations arranged anexpert work shop on implementation strategies for E-government in social security in the Autumn of 2000.This report summarizes the experts conclusions on strategies, methods, do´s and don´ts. It emphasizes theimportance of a mix of technological, political, legislational and organizational prerequisites.The considerations encompass the following domains or perspectives:(i) Infrastructure, being the technical devices such as network components, servers, protocols, instrumentsfor client identification, which needs some cooperation or coordination between social securityorganizations;(ii) Data management, which poses the question how governments can avoid to ask citizens or employersfor the same information twice;(iii) Standards and responsibilities, dealing with scope, and with how they are to be established,implemented and maintained;(iv) Client appreciation, one of the key issues when designing the services, which ones and how;(v) Issues of flexibility, which are related to changes in legislation, in technical standards and clientappreciation; and last but not least:(vi) Costs and benefits, the context of justification for investments.For each domain or theme, context, goals and experiences are stated first. Only a few examples aredescribed in the report itself. Each theme ends with do´s and don´ts, aiming at the promotion of action, atthe reduction not the ignorance of complexity. A range of illustrative cases is described in a separateappendix

Global Gene Expression Profiling of Individual Human Oocytes and Embryos Demonstrates Heterogeneity in Early Development

Early development in humans is characterised by low and variable embryonic viability, reflected in low fecundity and high rates of miscarriage, relative to other mammals. Data from assisted reproduction programmes provides additional evidence that this is largely mediated at the level of embryonic competence and is highly heterogeneous among embryos. Understanding the basis of this heterogeneity has important implications in a number of areas including: the regulation of early human development, disorders of pregnancy, assisted reproduction programmes, the long term health of children which may be programmed in early development, and the molecular basis of pluripotency in human stem cell populations. We have therefore investigated global gene expression profiles using polyAPCR amplification and microarray technology applied to individual human oocytes and 4-cell and blastocyst stage embryos. In order to explore the basis of any variability in detail, each developmental stage is replicated in triplicate. Our data show that although transcript profiles are highly stage-specific, within each stage they are relatively variable. We describe expression of a number of gene families and pathways including apoptosis, cell cycle and amino acid metabolism, which are variably expressed and may be reflective of embryonic developmental competence. Overall, our data suggest that heterogeneity in human embryo developmental competence is reflected in global transcript profiles, and that the vast majority of existing human embryo gene expression data based on pooled oocytes and embryos need to be reinterpreted

Global gene expression profiling of individual human oocytes and embryos demonstrates heterogeneity in early development

Early development in humans is characterised by low and variable embryonic viability, reflected in low fecundity and high rates of miscarriage, relative to other mammals. Data from assisted reproduction programmes provides additional evidence that this is largely mediated at the level of embryonic competence and is highly heterogeneous among embryos. Understanding the basis of this heterogeneity has important implications in a number of areas including: the regulation of early human development, disorders of pregnancy, assisted reproduction programmes, the long term health of children which may be programmed in early development, and the molecular basis of pluripotency in human stem cell populations. We have therefore investigated global gene expression profiles using polyAPCR amplification and microarray technology applied to individual human oocytes and 4-cell and blastocyst stage embryos. In order to explore the basis of any variability in detail, each developmental stage is replicated in triplicate. Our data show that although transcript profiles are highly stage-specific, within each stage they are relatively variable. We describe expression of a number of gene families and pathways including apoptosis, cell cycle and amino acid metabolism, which are variably expressed and may be reflective of embryonic developmental competence. Overall, our data suggest that heterogeneity in human embryo developmental competence is reflected in global transcript profiles, and that the vast majority of existing human embryo gene expression data based on pooled oocytes and embryos need to be reinterpreted

Chick tendon fibroblast transcriptome and shape depend on whether the cell has made its own collagen matrix

Collagen- and fibrin-based gels are extensively used to study cell behaviour. However, 2D-3D and collagen-fibrin comparisons of gene expression, cell shape and mechanotransduction, with an in vivo reference, have not been reported. Here we compared chick tendon fibroblasts (CTFs) at three stages of embryonic development with CTFs cultured in collagen- or fibrin-based tissue engineered constructs (TECs). CTFs synthesised their own collagen matrix in fibrin-based TECs and better recapitulated the gene expression, collagen fibril alignment and cell shape seen in vivo. In contrast, cells in 3D collagen gels exhibited a 2D-like morphology and expressed fewer of the genes expressed in vivo. Analysis of YAP/TAZ target genes showed that collagen gels desensitise mechanotransduction pathways. In conclusion, gene expression and cell shape are similar on plastic and 3D collagen whereas cells in 3D fibrin have a shape and transcriptome better resembling the in vivo situation. Implications for wound healing are discussed

PSR1 is a global transcriptional regulator of phosphorus deficiency responses and carbon storage metabolism in Chlamydomonas reinhardtii.

Many eukaryotic microalgae modify their metabolism in response to nutrient stresses such as phosphorus (P) starvation, which substantially induces storage metabolite biosynthesis, but the genetic mechanisms regulating this response are poorly understood. Here, we show that P starvation-induced lipid and starch accumulation is inhibited in a Chlamydomonas reinhardtii mutant lacking the transcription factor Pi Starvation Response1 (PSR1). Transcriptomic analysis identified specific metabolism transcripts that are induced by P starvation but misregulated in the psr1 mutant. These include transcripts for starch and triacylglycerol synthesis but also transcripts for photosynthesis-, redox-, and stress signaling-related proteins. To further examine the role of PSR1 in regulating lipid and starch metabolism, PSR1 complementation lines in the psr1 strain and PSR1 overexpression lines in a cell wall-deficient strain were generated. PSR1 expression in the psr1 lines was shown to be functional due to rescue of the psr1 phenotype. PSR1 overexpression lines exhibited increased starch content and number of starch granules per cell, which correlated with a higher expression of specific starch metabolism genes but reduced neutral lipid content. Furthermore, this phenotype was consistent in the presence and absence of acetate. Together, these results identify a key transcriptional regulator in global metabolism and demonstrate transcriptional engineering in microalgae to modulate starch biosynthesis

A revised bacterial polypeptide chain elongation cycle with a stepwise increase in restriction of unwanted ternary complexes by the ribosome

Metals in Catalysis, Biomimetics & Inorganic Material

Expulsion of Trichuris muris is associated with increased expression of angiogenin 4 in the gut and increased acidity of mucins within the goblet cell

<p>Abstract</p> <p>Background</p> <p><it>Trichuris muris </it>in the mouse is an invaluable model for infection of man with the gastrointestinal nematode <it>Trichuris trichiura</it>. Three <it>T. muris </it>isolates have been studied, the Edinburgh (E), the Japan (J) and the Sobreda (S) isolates. The S isolate survives to chronicity within the C57BL/6 host whereas E and J are expelled prior to reaching fecundity. How the S isolate survives so successfully in its host is unclear.</p> <p>Results</p> <p>Microarray analysis was used as a tool to identify genes whose expression could determine the differences in expulsion kinetics between the E and S <it>T. muris </it>isolates. Clear differences in gene expression profiles were evident as early as day 7 post-infection (p.i.). 43 probe sets associated with immune and defence responses were up-regulated in gut tissue from an E isolate-infected C57BL/6 mouse compared to tissue from an S isolate infection, including the message for the anti-microbial protein, angiogenin 4 (Ang4). This led to the identification of distinct differences in the goblet cell phenotype post-infection with the two isolates.</p> <p>Conclusion</p> <p>Differences in gene expression levels identified between the S and E-infected mice early during infection have furthered our knowledge of how the S isolate persists for longer than the E isolate in the C57BL/6 mouse. Potential new targets for manipulation in order to aid expulsion have been identified. Further we provide evidence for a potential new marker involving the acidity of the mucins within the goblet cell which may predict outcome of infection within days of parasite exposure.</p

Human notochordal cell transcriptome unveils potential regulators of cell function in the developing intervertebral disc

The adult nucleus pulposus originates from the embryonic notochord, but loss of notochordal cells with skeletal maturity in humans is thought to contribute to the onset of intervertebral disc degeneration. Thus, defining the phenotype of human embryonic/fetal notochordal cells is essential for understanding their roles and for development of novel therapies. However, a detailed transcriptomic profiling of human notochordal cells has never been achieved. In this study, the notochord-specific marker CD24 was used to specifically label and isolate (using FACS) notochordal cells from human embryonic and fetal spines (7.5-14 weeks post-conception). Microarray analysis and qPCR validation identified CD24, STMN2, RTN1, PRPH, CXCL12, IGF1, MAP1B, ISL1, CLDN1 and THBS2 as notochord-specific markers. Expression of these markers was confirmed in nucleus pulposus cells from aged and degenerate discs. Ingenuity pathway analysis revealed molecules involved in inhibition of vascularisation (WISP2, Noggin and EDN2) and inflammation (IL1-RN) to be master regulators of notochordal genes. Importantly, this study has, for the first time, defined the human notochordal cell transcriptome and suggests inhibition of inflammation and vascularisation may be key roles for notochordal cells during intervertebral disc development. The molecules and pathways identified in this study have potential for use in developing strategies to retard/prevent disc degeneration, or regenerate tissue.This research was funded by Arthritis Research UK (reference 21165 to SMR/JAH/LW) and the Henry Smith Charity (to JAH/SMR/MH). RRP was supported by a grant from the Programme for Advanced Medical Education, sponsored by Fundacão Calouste Gulbenkian, fundação Champalimaud, Ministério da Saúde, Fundação para a Ciência e Tecnologia and Apifarma, Portugal. Support was also received from the UK Medical Research Council (MR/J003352/1 to KPH), the Wellcome Trust (NAH was a senior fellow in clinical science, 088566; additional support from grant, 097820), and the British Council (14BX15NHBG to NAH). Consumable and technical support for this project (Sonal Patel) was funded by the National Institute for Health Research Manchester Musculoskeletal Biomedical Research Unit. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the National Institute for Health Research, or the Department of Health. We are very grateful to all women who consented to take part in our research programme and for the assistance of research nurses and clinical colleagues at Manchester University NHS Foundation Trust. The authors would like to thank Prof Kathy Cheah and group for advice regarding the identification of sex-specific markers within microarray datasets.info:eu-repo/semantics/publishedVersio

WT1 and its transcriptional cofactor BASP1 redirect the differentiation pathway of an established blood cell line

The Wilms' tumour suppressor WT1 (Wilms' tumour 1) is a transcriptional regulator that plays a central role in organogenesis, and is mutated or aberrantly expressed in several childhood and adult malignancies. We previously identified BASP1 (brain acid-soluble protein 1) as a WT1 cofactor that suppresses the transcriptional activation function of WT1. In the present study we have analysed the dynamic between WT1 and BASP1 in the regulation of gene expression in myelogenous leukaemia K562 cells. Our findings reveal that BASP1 is a significant regulator of WT1 that is recruited to WT1-binding sites and suppresses WT1-mediated transcriptional activation at several WT1 target genes. We find that WT1 and BASP1 can divert the differentiation programme of K562 cells to a non-blood cell type following induction by the phorbol ester PMA. WT1 and BASP1 co-operate to induce the differentiation of K562 cells to a neuronal-like morphology that exhibits extensive arborization, and the expression of several genes involved in neurite outgrowth and synapse formation. Functional analysis revealed the relevance of the transcriptional reprogramming and morphological changes, in that the cells elicited a response to the neurotransmitter ATP. Taken together, the results of the present study reveal that WT1 and BASP1 can divert the lineage potential of an established blood cell line towards a cell with neuronal characteristics