The power of network-based drug design and the interplay between metabolism and gene expression in Trypanosoma brucei

Westerhoff, H.V. [Promotor]Bakker, B.M. [Copromotor

Nurturing Neighbourhoods: Parent Outreach Worker Program, Guelph, Ontario.

The "Parent Outreach Worker Program" (POW program) aims to identify, reach out to, and assist families with children who may be isolated or otherwise struggling by providing them with increased access to services and support. The POW program is currently being piloted in two priority neighbourhoods of Guelph, Ontario. The POW program is part of a cross-sectoral, community-based "Nurturing Neighbourhoods" initiative that is grounded in the philosophies and principles related to the social determinants of health, health equity, and early help in child welfare. This report details the findings of an evaluation that was conducted in 2013-2014 to determine whether the POW program is being delivered as planned and the extent to which it is achieving its intended outcomes. Evidence used for the evaluation include program implementation data inputted by the Parent Outreach Workers (POWs) and a series of interviews and focus groups conducted with program stakeholders including the POWs, parents who use the POW program, and a variety of associated service providers and community resource people. This early evaluation evidence indicates clearly that the POW program is generating very significant outcomes for priority families as direct supports and as intermediaries between those families and the community resources they need. The POWs occupy an important space in marginalized communities, the evaluation evidence shows, between formal services, informal community supports, and the families most in need of support. Also emerging from the evaluation data are some key "Impact Pathways" through which the POW program is generating results. These Impact Pathways reflect many of the strategies found to be effective in research literature on other outreach programs, and as such may serve as a best practices framework for the establishment of similar programs in other neighbourhoods.Guelph Community Health Centre, Wellington-Dufferin-Guelph Public Health, Family & Children's Services of Guelph and Wellington Coun

Nurturing Neighbourhoods: Parent Outreach Worker Program, Guelph, Ontario. Evaluation Report: November 2014

The "Parent Outreach Worker Program" (POW program) aims to identify, reach out to, and assist families with children who may be isolated or otherwise struggling by providing them with increased access to services and support. The POW program is currently being piloted in two priority neighbourhoods of Guelph, Ontario. The POW program is part of a cross-sectoral, community-based "Nurturing Neighbourhoods" initiative that is grounded in the philosophies and principles related to the social determinants of health, health equity, and early help in child welfare. This report details the findings of an evaluation that was conducted in 2013-2014 to determine whether the POW program is being delivered as planned and the extent to which it is achieving its intended outcomes. Evidence used for the evaluation include program implementation data inputted by the Parent Outreach Workers (POWs) and a series of interviews and focus groups conducted with program stakeholders including the POWs, parents who use the POW program, and a variety of associated service providers and community resource people. This early evaluation evidence indicates clearly that the POW program is generating very significant outcomes for priority families as direct supports and as intermediaries between those families and the community resources they need. The POWs occupy an important space in marginalized communities, the evaluation evidence shows, between formal services, informal community supports, and the families most in need of support. Also emerging from the evaluation data are some key "Impact Pathways" through which the POW program is generating results. These Impact Pathways reflect many of the strategies found to be effective in research literature on other outreach programs, and as such may serve as a best practices framework for the establishment of similar programs in other neighbourhoods

Explicit consideration of topological and parameter uncertainty gives new insights into a well-established model of glycolysis

Previous models of glycolysis in the sleeping sickness parasite Trypanosoma brucei assumed that the core part of glycolysis in this unicellular parasite is tightly compartimentalized within an organelle, the glycosome, which had previously been shown to contain most of the glycolytic enzymes. The glycosomes were assumed to be largely impermeable, and exchange of metabolites between the cytosol and the glycosome was assumed to be regulated by specific transporters in the glycosomal membrane. This tight compartmentalization was considered to be essential for parasite viability. Recently, size-specific metabolite pores were discovered in the membrane of glycosomes. These channels are proposed to allow smaller metabolites to diffuse across the membrane but not larger ones. In light of this new finding, we re-analyzed the model taking into account uncertainty about the topology of the metabolic system in T. brucei, as well as uncertainty about the values of all parameters of individual enzymatic reactions. Our analysis shows that these newly-discovered nonspecific pores are not necessarily incompatible with our current knowledge of the glycosomal metabolic system, provided that the known cytosolic activities of the glycosomal enzymes play an important role in the regulation of glycolytic fluxes and the concentration of metabolic intermediates of the pathway

Design principles for successful adoption of life cycle thinking in asset management decision-making

While Asset Management (AM) organizations are increasingly using Life Cycle Thinking (LCT) studies to assess their long-term plans, the translation of the LCT outcomes into actionable decisions often appears to be lacking. The international industry standard on AM can aid in improving organizational sustainability and is therefore used to study fourteen LCT cases. Subsequently, several design criteria and principles are suggested, based on existing literature. Clarity about ownership is necessary in order to align the perspectives of project leaders and LCT performers. Furthermore, attention should be distributed across the analysis and subsequent action, and better-fitting management styles should be applied

Costimulation blockade and regulatory T-cells in a non-human primate model of kidney allograft transplantation

Successful tolerance induction therapies in rodents are for the most part unsuccessful in larger primates. Costimulation blockade by anti-CD40 or anti-CD40 + anti-CD86 in the life-supporting kidney allograft model in the rhesus monkey prevented graft rejection during treatment but did not induce tolerance. Costimulation blockade has to be integrated in conventional immunosuppressive therapies. However, conventional immunosuppressive drugs may antagonize the immunoregulatory effects of costimulation blockade. We describe how costimulation blockade prevented graft rejection in the immediate post transplantation period. CsA treatment was initiated only after 42 days post transplantation and this had a beneficial effect on graft survival, resulting in two of 4 monkeys surviving long-term without additional immunosuppressive treatment. Furthermore we found no beneficial effect of ATG induction therapy on costimulation blockade treatment. ATG induced rapid reappearance of CD8+ memory T-cells in the peripheral blood, possibly responsible for the observed accelerated rejection. Infiltrating cells in kidney graft biopsies and tissues revealed high expression of FOXP3 and other regulatory T-cell markers during rejection. We also have described the phenotypic and functional characteristics of naturally occurring regulatory T-cells in rhesus monkeys.Dutch Kidney Foundation (Nierstichting) Dutch Transplantation Society (NTV) PanGenetics BV BPRCUBL - phd migration 201

Characterization of resistance genes to Cladosporium fulvum on the short arm of chromosome 1 of tomato

Plant breeders generally use qualitative resistance that is associated with a hypersensitive reaction (HR) to obtain cultivars that are resistant to pathogens and pests. The genetics of this resistance is based on the gene-for-gene relationship, which involves the product of a plant resistance gene and the product of an avirulence gene of the pathogen occurs. The interaction between leaf mold ( Cladosporium fulvum ) and its solely host, tomato ( Lycopersicon esculentum ), complies with this model. In the last few years, the isolation of several avirulence ( Avr ) and resistance ( R ) genes have contributed to an increase in our knowledge on this interaction. Several resistance genes to C. fulvum ( Cf genes) have been isolated from tomato. These Cf genes are located on two different clusters on the tomato genome, which contain not only functional Cf genes, but also several homologs with yet unknown function.. The short arm of Chromosome 1 harbors one of these clusters, designated "Milky Way", comprising functional Cf genes ( Cf-4 , Cf-4A , Cf-9 ). Moreover, two other clusters are located on the short arm of Chromosome 1, designated "Northern Lights" and "Southern Cross", which only harbor homologs ( Hcr9 s), but no functional Cf genes. Also, there are several reports about the presence of other Cf genes on the short arm of Chromosome 1.To increase our knowledge on the genetic and molecular organization of Cf genes on the short arm of Chromosome 1, an experimental approach was chosen to identify Cf genes with novel specificities that map on the short arm of Chromosome 1. To saturate the tomato genome with molecular markers, an integrated high-density AFLP-RFLP map was constructed using two different L. esculentum x L. pennellii F 2 mapping populations. Although 1175 AFLP markers were mapped on the tomato genome, covering 1482 cM, only nine AFLP markers were detected between the RFLP markers CT233 and TG51, which mark a 23.6 cM interval, comprising several Hcr9 clusters, on the short arm of Chromosome 1. This relatively low number of markers is due to the clustering of most Eco RI/ Mse I AFLP markers around the centromeres (Chapter 2).Testcross populations of 66 C. fulvum resistant Lycopersicon accessions were obtained by crossing these accessions with the near isogenic line Moneymaker-Cf4 and subsequent crossing of the F 1 with the susceptible tomato cultivar Moneymaker (Chapter 3). Using disease resistance tests with C. fulvum race 0 on only 24 plants of these testcross populations, susceptible plants were identified. An under-representation of susceptible plants identified Cf resistance linked to Cf-4 , and hence location of the unknown resistance on the short arm of Chromosome 1. Out of the 21 resistant accessions that have been tested in this way, ten showed a Cf-4 linked Cf gene. Among these ten accessions, one accession specifically recognized the extracellular protein ECP5 of C. fulvum and the corresponding gene was designated Cf-ECP5 . This gene was more accurately mapped using a testcross population of 338 plants and an F 2 population of a cross between Moneymaker and CfECP5, consisting of 233 individuals. Cf-ECP5 mapped 4 cM proximal to the Hcr9 locus Milky Way and the corresponding Cf locus was designated Aurora. An amplification product that cosegregated completely with the Cf-ECP5 gene, was cloned and nine clones were sequenced (Chapter 6). These nine clones could be classified into four groups, indicating that the Aurora locus comprises several Hcr9 s.Of the 66 resistant Lycopersicon accessions mentioned above, 64 have been screened for the presence of Cf-4 and/or Cf-9 , using PVX:: Avr4 and PVX:: Avr9 , respectively. A relatively large proportion of these accessions all harbored the functional genes Cf-4 and Cf-4A (Chapter 4). Sequence analysis of the 3' end of Cf-4 and part of the 3' untranslated region of Cf-4 showed no differences from the previously published Cf-4 sequences, hence these lines contain an introgression fragment with identical Cf-4 and Cf-4A genes. Since several of these lines were previously designated with different Cf digits, a change in nomenclature is proposed.Five out of the 66 accessions studied, showed an HR upon specific recognition of ECP2 and therefore harbor the corresponding resistance gene Cf-ECP2 (Chapter 5). Using two different testcross populations and one F 2 population from a cross between Moneymaker and CfECP2, representing in total 282 individuals, Cf-ECP2 was accurately mapped. Cf-ECP2 cosegregates with the molecular marker CT116, which is located proximal to the Milky Way and Aurora clusters, but distal to the Southern Cross locus. Southern hybridization, using Cf-9 as a probe, showed a hybridizing band of7.5 kb cosegregating with Cf-ECP2 , indicating that Cf-ECP2 is a member of a previously unidentified Hcr9 locus, that has been designated Orion.Studies in Chapters 3, 5 and 6 show that functional Cf genes can be located on several different Hcr9 loci on the short arm of Chromosome 1 and that these Hcr9 loci are highly polymorphic.</p