Interleukin-4-mediated regulation of CD25 gene expression in human B lymphocytes

This thesis describes the regulation of CD25 gene expression by interleukin-4-activated signal transduction pathways in quiescent human B lymphocytes. In particular, the importance of a cAMP-dependent pathway, which down-regulates binding of a putative transcriptionally repressive protein factor from a negative regulatory element in the promoter region of the CD25 gene, is highlighted. CD25 is the alpha chain of the multi- component interleukin-2 receptor complex and is a requirement for formation of high affinity interleukin-2 receptors. In addition, CD25 is the only unique component of the interleukin-2 receptor and as such it confers rigourous specificity for binding of interleukin-2. To date, interleukui-4 is the only cytokine recognized to be capable of upregulating CD25 in human B lymphocytes. The initial aim of this work was to identify signalling pathways which were responsible for up-regulation of CD25 expression, by artificially activating specific second messenger systems. These early studies were based on the limited knowledge of interleukin-4-induced signal transduction pathways available at the time; treatment of resting human B cells with phorbol ester, calcium ionophore and forskolin results in modest increases in cell surface levels of CD25. Additionally, these stimuli and those required for elevation of surface CD23 levels are similar. It seemed possible therefore, that the ability of IL-4 to promote increases in intracellular calcium, protein kinase C activation and cAMP generation might have a role in IL-4-induced CD25 expression. Additionally, chelation of intracellular calcium and down-regulation of PKC, by chronic phorbol ester treatment, attenuates the ability of IL-4 to up-regulate CD25. Similarly, this latter treatment abrogates any increase in CD25 levels observed in response to anti-Iimmunoglobulin and anti-CD40 antibodies, the only other stimuli capable of inducing CD25 expression in primary human B cells. Upon binding to its receptor, IL-4 induces a biphasic elevation of intracellular cAMP; a transient increase which peaks at approximately 2 minutes and is followed by a more sustained increase after 10 minutes. This sustained cAMP generation is maintained until approximately 30 minutes after stimulation, before dropping back to basal levels. Additionally, we have demonstrated that IL-4-induced cAMP production is dose dependent and leads to the activation of the cAMP-dependent protein kinase, PKA, in primary human B cells. (Abstract shortened by ProQuest.)

Assessing occupational participation among justice involved people ‘with a personality disorder’: Quantitative assessments and their properties

Introduction: There is little evidence for what influences occupational participation for justice-involved people ‘with a personality disorder’ living in community contexts, and no validated occupational participation assessments specific to this group. We assessed a sample of justice-involved people ‘with a personality disorder’ to ascertain what influences occupational participation using commonly applied assessments and evaluated their construct validity. Method: As part of a mixed-methods study, a purposive sample of 18 justice-involved people ‘with a personality disorder’ were scored on the Model of Human Occupational Screening Tool and Occupational Performance History Interview–Version Two scales. Mean scores were calculated per Model of Human Occupational Screening Tool (MOHOST) item and Occupational Performance History Interview–Version Two (OPHI-II) items and scales and compared to published data. Mann–Whitney U Tests were used to identify within-sample differences based on demographic characteristics. Results: Participants had low scores on MOHOST items and OPHI-II items and scales. Differences were identified compared to published data. Within-sample differences were most apparent in comparisons by employment status and ethnicity. The OPHI-II scales did not operate as intended with this population and recommended adjustments impacted its construct validity. Conclusion: Replication is required with a larger random sample. Integrating these data with qualitative exploration would further elucidate factors influencing occupational participation in this population

How effective are interventions to improve social outcomes among offenders with personality disorder: a systematic review

Background: Offenders with personality disorder are supported by health, criminal justice, social care and third sectorservices. These services are tasked with reducing risk, improving health and improving social outcomes. Research hasbeen conducted into interventions that reduce risk or improve health. However, interventions to improve social outcomesare less clearly defined.Methods: To review the effectiveness of interventions to improve social outcomes we conducted a systematic reviewusing Cochrane methodology, expanded to include non-randomised trials. Anticipated high heterogeneity of the studiesinformed narrative synthesis.Results: Eleven studies met inclusion criteria. Five contained extractable data. No high-quality studies were identified.Outcomes measured clustered around employment and social functioning. Interventions vary and their mechanismsfor influencing social outcomes are poorly operationalised. Although change was observed in employment rates, therewas no evidence for the effectiveness of these interventions.Conclusions: There is a lack of evidence for effective interventions that improve social outcomes. Further research isrecommended to reach consensus on the outcomes of importance, identify the factors that influence these and designtheoretically-informed and evidence-based interventions

Developing an Intervention to Improve Occupational Participation for Justice-Involved People with a Personality Disorder: Defining and Describing Intervention Components

Occupational participation is undertaking personally meaningful and socially valued activities and roles. It is an important outcome for health and justice interventions, as it is integral to health and desistance. We report the third of a four-stage research project to develop an intervention to improve occupational participation for justice-involved people with a personality disorder in the community. We completed a Delphi survey to produce expert consensus on intervention components and their content, ascertain participant ratings of 28 factors for their level of influence on occupational participation, and the modifiability of the factors with this population. Thirty multi-disciplinary participants completed three survey rounds. Most factors were rated very influential, but few were considered easily modifiable. Participants agreed 121 statements describing intervention components and content. Twenty-seven statements did not reach consensus. In targeting specific factors in intervention, practitioners must balance their degree of influence with potential modifiability. The results will inform intervention manualization and modeling

SOCS3 Is Essential in the Regulation of Fetal Liver Erythropoiesis

AbstractSOCS3 (CIS3/JAB2) is an SH2-containing protein that binds to the activation loop of Janus kinases, inhibiting kinase activity, and thereby suppressing cytokine signaling. During embryonic development, SOCS3 is highly expressed in erythroid lineage cells and is Epo independent. Transgene-mediated expression blocks fetal erythropoiesis, resulting in embryonic lethality. SOCS3 deletion results in an embryonic lethality at 12–16 days associated with marked erythrocytosis. Moreover, the in vitro proliferative capacity of progenitors is greatly increased. SOCS3-deficient fetal liver stem cells can reconstitute hematopoiesis in lethally irradiated adults, indicating that its absence does not disturb bone marrow erythropoiesis. Reconstitution of lymphoid lineages in JAK3-deficient mice also occurs normally. The results demonstrate that SOCS3 is critical in negatively regulating fetal liver hematopoiesis

Enhanced RAD21 cohesin expression confers poor prognosis and resistance to chemotherapy in high grade luminal, basal and HER2 breast cancers

Introduction: RAD21 is a component of the cohesin complex, which is essential for chromosome segregation and error-free DNA repair. We assessed its prognostic and predictive power in a cohort of in situ and invasive breast cancers, and its effect on chemosensitivity in vitro.Methods: RAD21 immunohistochemistry was performed on 345 invasive and 60 pure in situ carcinomas. Integrated genomic and transcriptomic analyses were performed on a further 48 grade 3 invasive cancers. Chemosensitivity was assessed in breast cancer cell lines with an engineered spectrum of RAD21 expression.Results: RAD21 expression correlated with early relapse in all patients (hazard ratio (HR) 1.74, 95% confidence interval (CI) 1.06 to 2.86, P = 0.029). This was due to the effect of grade 3 tumors (but not grade 1 or 2) in which RAD21 expression correlated with early relapse in luminal (P = 0.040), basal (P = 0.018) and HER2 (P = 0.039) groups. In patients treated with chemotherapy, RAD21 expression was associated with shorter overall survival (P = 0.020). RAD21 mRNA expression correlated with DNA copy number, with amplification present in 32% (7/22) of luminal, 31% (4/13) of basal and 22% (2/9) of HER2 grade 3 cancers. Variations in RAD21 mRNA expression in the clinical samples were reflected in the gene expression data from 36 breast cancer cell lines. Knockdown of RAD21 in the MDA-MB-231 breast cancer cell line significantly enhanced sensitivity to cyclophosphamide, 5-fluorouracil and etoposide. The findings for the former two drugs recapitulated the clinical findings.Conclusions: RAD21 expression confers poor prognosis and resistance to chemotherapy in high grade luminal, basal and HER2 breast cancers. RAD21 may be a novel therapeutic target

Advance Choices, and Medical Decision Making in Intensive Care Situations : Paper by the Law Society of Scotland

[We offer a basic formulation (previously lacking) of the doctor-patient relationship, and resulting obligations, responsibilities and potential liabilities, in any situation where medical decision-making cannot proceed at all, or sufficiently quickly, in accordance with the competent and informed consent of the patient. The formulation is derived from existing principles in Scots law and some similar legal systems. Adequate clarity and certainty are however at present lacking in Scots law. It is urgently necessary to provide it, in the interests of doctors, patients and all others who might be concerned. In the matter of withholding or withdrawing life-sustaining treatment, we have considered, commented upon and evaluated the 1995 proposals by Scottish Law Commission (which were never implemented), together with such case law as has developed, as a basis for formulating legislative provision that would provide the clarity, certainty and protections for medical practitioners acting properly, that is at present lacking in Scotland. Our recommendations include suggestions as to how both topics may be taken forward to the drafting, introduction and implementation of legislation, including suggestions as to further research, consultation, and – following upon the introduction of legislation – a coordinated approach to all aspects of its successful implementation.

Polymorphisms in a Putative Enhancer at the 10q21.2 Breast Cancer Risk Locus Regulate NRBF2 Expression.

Genome-wide association studies have identified SNPs near ZNF365 at 10q21.2 that are associated with both breast cancer risk and mammographic density. To identify the most likely causal SNPs, we fine mapped the association signal by genotyping 428 SNPs across the region in 89,050 European and 12,893 Asian case and control subjects from the Breast Cancer Association Consortium. We identified four independent sets of correlated, highly trait-associated variants (iCHAVs), three of which were located within ZNF365. The most strongly risk-associated SNP, rs10995201 in iCHAV1, showed clear evidence of association with both estrogen receptor (ER)-positive (OR = 0.85 [0.82-0.88]) and ER-negative (OR = 0.87 [0.82-0.91]) disease, and was also the SNP most strongly associated with percent mammographic density. iCHAV2 (lead SNP, chr10: 64,258,684:D) and iCHAV3 (lead SNP, rs7922449) were also associated with ER-positive (OR = 0.93 [0.91-0.95] and OR = 1.06 [1.03-1.09]) and ER-negative (OR = 0.95 [0.91-0.98] and OR = 1.08 [1.04-1.13]) disease. There was weaker evidence for iCHAV4, located 5' of ADO, associated only with ER-positive breast cancer (OR = 0.93 [0.90-0.96]). We found 12, 17, 18, and 2 candidate causal SNPs for breast cancer in iCHAVs 1-4, respectively. Chromosome conformation capture analysis showed that iCHAV2 interacts with the ZNF365 and NRBF2 (more than 600 kb away) promoters in normal and cancerous breast epithelial cells. Luciferase assays did not identify SNPs that affect transactivation of ZNF365, but identified a protective haplotype in iCHAV2, associated with silencing of the NRBF2 promoter, implicating this gene in the etiology of breast cancer.This is the author accepted manuscript. The final version is available from Elsevier via http://dx.doi.org/10.1016/j.ajhg.2015.05.002

Evidence that breast cancer risk at the 2q35 locus is mediated through IGFBP5 regulation.

GWAS have identified a breast cancer susceptibility locus on 2q35. Here we report the fine mapping of this locus using data from 101,943 subjects from 50 case-control studies. We genotype 276 SNPs using the 'iCOGS' genotyping array and impute genotypes for a further 1,284 using 1000 Genomes Project data. All but two, strongly correlated SNPs (rs4442975 G/T and rs6721996 G/A) are excluded as candidate causal variants at odds against >100:1. The best functional candidate, rs4442975, is associated with oestrogen receptor positive (ER+) disease with an odds ratio (OR) in Europeans of 0.85 (95% confidence interval=0.84-0.87; P=1.7 × 10(-43)) per t-allele. This SNP flanks a transcriptional enhancer that physically interacts with the promoter of IGFBP5 (encoding insulin-like growth factor-binding protein 5) and displays allele-specific gene expression, FOXA1 binding and chromatin looping. Evidence suggests that the g-allele confers increased breast cancer susceptibility through relative downregulation of IGFBP5, a gene with known roles in breast cell biology

Genome-wide association analysis of more than 120,000 individuals identifies 15 new susceptibility loci for breast cancer.

Genome-wide association studies (GWAS) and large-scale replication studies have identified common variants in 79 loci associated with breast cancer, explaining ∼14% of the familial risk of the disease. To identify new susceptibility loci, we performed a meta-analysis of 11 GWAS, comprising 15,748 breast cancer cases and 18,084 controls together with 46,785 cases and 42,892 controls from 41 studies genotyped on a 211,155-marker custom array (iCOGS). Analyses were restricted to women of European ancestry. We generated genotypes for more than 11 million SNPs by imputation using the 1000 Genomes Project reference panel, and we identified 15 new loci associated with breast cancer at P < 5 × 10(-8). Combining association analysis with ChIP-seq chromatin binding data in mammary cell lines and ChIA-PET chromatin interaction data from ENCODE, we identified likely target genes in two regions: SETBP1 at 18q12.3 and RNF115 and PDZK1 at 1q21.1. One association appears to be driven by an amino acid substitution encoded in EXO1.BCAC is funded by Cancer Research UK (C1287/A10118, C1287/A12014) and by the European Community's Seventh Framework Programme under grant agreement 223175 (HEALTH-F2-2009-223175) (COGS). Meetings of the BCAC have been funded by the European Union COST programme (BM0606). Genotyping on the iCOGS array was funded by the European Union (HEALTH-F2-2009-223175), Cancer Research UK (C1287/A10710, C8197/A16565), the Canadian Institutes of Health Research (CIHR) for the CIHR Team in Familial Risks of Breast Cancer program and the Ministry of Economic Development, Innovation and Export Trade of Quebec, grant PSR-SIIRI-701. Combination of the GWAS data was supported in part by the US National Institutes of Health (NIH) Cancer Post-Cancer GWAS initiative, grant 1 U19 CA148065-01 (DRIVE, part of the GAME-ON initiative). For a full description of funding and acknowledgments, see the Supplementary Note.This is the author accepted manuscript. The final version is available from NPG via http://dx.doi.org/10.1038/ng.324