Dataset Information

Health Service Intervention for the Improvement of Access and Adherence to Colorectal Cancer Screening

ABSTRACT: This phase I study investigates how well a health service program called Witness CARES Services works in increasing access and adherence to colorectal cancer screening in patients who have not undergone a colorectal screening in the past year. Providing patients with navigational and screening services via Witness CARES Services may improve access to and adherence levels of colorectal screening.

DISEASE(S): Colorectal Carcinoma,Colorectal Neoplasms

PROVIDER: 2356048 | ecrin-mdr-crc |

REPOSITORIES: ECRIN MDR

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Project description:Food-borne illness arising for Shiga-toxigenic Escherichia coli is often linked to consumption of fruit and vegetables as the bacteria have the ability to interact with plants and use them as alternative or secondary hosts. Attachment of the bacteria to host tissue is one of the first steps in the interaction, and, as with mammalian hosts, has shown to be mediated by a combination of non-specific and specific adhesin-mediated interactions. We took a high-throughput positive-selection approach to investigate adherence mechanisms for E. coli O157:H7 isolate Sakai by inoculating a BAC clone library onto spinach, which was quantified by microarray hybridisation and gene loci enrichment measured using a Bayesian hierarchical model. The screen involved four successive rounds of adherence to spinach roots, resulting in 115 CDS credible candidates, covered by seven contiguous genomic regions. Two candidates regions selected for functional assessment included a chaperone-usher fimbrial gene cluster (loc6) and the type two secretion system (T2SS). The TS22 was found to significantly enhance binding to spinach roots and leaves, demonstrated with a BAC-T2SS clone and by mutagenesis of the secretin protein, EtpD. Both etpD and the inner membrane anchor protein gene etpC were expressed at 18 degree celsius, and expression of etpD was demonstrated for STEC (Sakai) resident in the apoplastic spaces in spinach leaf tissue. Together, these data indicate a novel function for STEC T2SS in adherence to plant tissue. Experiment 1: screening E coli O157:H7 Sakai genes for adherence to spinach roots. A BAC library of Sakai clones in an E coli DH10B background (which has poor root adherence) defined as the 'Input pool', was incubated with spinach roots for 4 rounds of enrichment, defined as the 'Output pool'. Control samples (defined as 'Input control' & 'Output control') were cultures of pV41 vector only. DNA extractions from test pools were labelled with Cy3 throughout. DH10B DNA was used for grid alignment and labelled with Cy5 throughout.

Project description:We developed spotted oligonucleotide arrays of the S. pyogenes SF370 (an M1 serotype) genome and compared the transcriptomes of streptococci that adhere to Detroit 562 human pharyngeal cells to non-adherent (“associated”) streptococci within the same experiment. We replicated experiments independently and used dye-swaps to incorporate biological and technical variation, biological replicate experiments. Following filtering and normalization, we analyzed data from four biological replicates with robust summary statistics, Bayesian statistics and permutation algorithms to identify genes differentially expressed with significance during pharyngeal cell adherence. This analysis identified 79 genes (4% of the genome) exhibiting statistically significant fold changes in expression (PF value < 0.05) during adherence from 1769 open reading frames represented on the array. The genes encoding several known virulence factors (e.g. cysteine protease SpeB and streptolysin O) were differentially expressed with significance; however, many were either never before associated with adherence (e.g. phage-encoded genes including speH), or of undefined function. Genes demonstrating up-regulation (n= 45) and down-regulation (n= 34) included virulence factors, prophage-encoded transcripts, metabolic genes and transcriptional regulators (Table 2). Undefined or hypothetical genes comprised 27% of differentially expressed genes (n= 21; 11 chromosomally-encoded, 10 phage-encoded). We conducted TaqMan (qRT-PCR) analysis of 11 differentially expressed genes to validate selected microarray hybridization results. We further analyzed the data with neighbor clustering, using a set of novel computational algorithms (termed “GenomeCrawler”) for evaluating bacterial microarray data, which couples the expression profiles of genes with their physical location on the chromosome. When applied to the microarray data from this study, neighbor clustering identified a greater number of differentially expressed genes, which facilitated the reconstruction of known multimeric proteins, complete metabolic pathways and intact virulence loci that would not have been possible without its application. These data sets and algorithms are freely available for download at www.rockefeller.edu/vaf. Keywords: Pharyngeal cell adherent vs. associated

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Health Service Intervention for the Improvement of Access and Adherence to Colorectal Cancer Screening

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