Developing peptide vaccines for breast cancer

The complex and heterogeneous nature of breast cancer presents significant challenges for many conventional cancer therapies. Immunotherapy has the potential to provide a more potent, less invasive and less toxic approach to breast cancer treatment, with several successes already evident in the treatment of metastatic melanoma. Three recently described tumour-associated antigens (TAAs); (1) NY-BR-1, (2) cadherin-3/P-cadherin (CDH3), and (3) bone marrow stromal cell antigen-2 (BST-2), have been implicated as candidates for breast cancer vaccination strategies. However, peptide vaccines that are designed using wild-type (WT) sequences of these proteins are likely to elicit suboptimal T-cell responses in patients. This is generally attributable to the low affinity of thymically-selected Tcell receptors (TCRs) for “self” TAAs. Here I describe a method for enhancing the immunogenicity of these three HLA-A*0201 (HLA-A2)-restricted breast cancer epitopes. Firstly, αβ CD8+ T-cell clones were generated against each of the epitopes using three different approaches; (i) T-cell lines and (ii) T-cell libraries (Chapter 3),and (iii) tumour-infiltrating lymphocytes (TILs) (Chapter 4), and the breast cancer reactivity of these clones confirmed in vitro. Then, using a positional scanning synthetic combinatorial library (PS-SCL), altered peptide ligands (APLs) were designed for each of the clones (Chapter 5). These APLs were shown to be up to 100,000-fold more potent than the respective WT epitopes. Preliminary “proof-ofconcept” CD8+ T-cell priming experiments on healthy HLA-A2+ donors were then used to establish whether a chosen BST-2 APL was capable of activating superior breast cancer specific T-cell populations in vitro (Chapter 6). From these preliminary investigations, it was found that the BST-2 APL was capable of generating a T-cell response of greater magnitude, and that was also better able to kill breast cancer cells when compared to the corresponding BST-2 WT peptide. Together, these experiments illustrate the potential use of APLs for the development of a highly effective prophylactic or therapeutic peptide vaccine for breast cancer

Assessment of patients’ self-reported levels of adherence to postoperative restrictions following total hip replacement

Background: Postoperative precautions that limit hip movement reduce the incidence of postoperative dislocation following total hip replacement (THR). It is assumed that patients adhere to these precautions, but true adherence is unknown. Aims: To assess adherence to postoperative precautions, including bending, twisting, crossing of legs, sleeping position and sitting position following primary THR surgery. Methods: A self-reporting questionnaire explored patient adherence to precautions following primary THR. Questionnaires were sent to 120 patients following elective primary THR at two orthopaedic centres in England between November 2016 and April 2017. Patients were also asked to report the duration of adherence and the difficulty associated with adherence to each of the precautions. Results: Eighty-three percent (99/120) of patients responded. Of these, 56% (56/99) were female and 44% (44/99) were male. Mean age was 66.0 (standard deviation 9.4) years. Seventy-six percent of patients were classified as ‘highly adherent’ to the precautions in hospital, and this reduced to 68% when patients went home. The median duration of adherence to the precaution to avoid crossing legs was 6 weeks [interquartile range (IQR) 6–6]. For the remaining four precautions, the median duration of adherence was consistently 6 weeks (IQR 4–6). Conclusion: High levels of adherence to precautions following THR were identified. However, patients reported difficulty adhering to the precautions, which may explain why 24% of patients were not able to adhere to the precautions> 90% of the time. Patients reported that the most difficult precaution to adhere to was supine sleeping, and the least difficult precautions to adhere to were avoiding crossing legs and using a raised chair

Borneo stalagmite evidence of significantly reduced El Niño-Southern Oscillation variability at 4.1 kyBP

The timing and geographic extent of a potential “4.2 ky event” remain highly contested. Here we present records of ENSO variability at 3.8 kyBP and 4.1 kyBP derived from a Borneo stalagmite, which suggest a significant change in ENSO properties between these time intervals. The Borneo records show evidence of significantly reduced ENSO activity at 4.1 kyBP, relative to other measured windows within the Holocene. This reduced ENSO activity coincides with a period of drier conditions and enhanced dust events in the Middle East that took place ∼4.0–4.3 kyBP. The Borneo records show evidence of enhanced ENSO activity at 3.8 kyBP. Various hydroclimate changes attributed to the “4.2 ky event” in many regions may thus be reflecting a shift from reduced to enhanced El Niño activity that occurred between 3.8 kyBP to 4.0 kyBP

Sceptical Employees as CSR Ambassadors in Times of Financial Uncertainty

This chapter offers new insights into the understanding of internal (employee) perceptions of organizational corporate social responsibility (CSR) policies and strategies. This study explores the significance of employees’ involvement and scepticism upon CSR initiatives and focuses on the effects it may have upon word of mouth (WOM) and the development of employee–organisation relationships. Desk research introduces the research questions. Data for the research questions were gathered through a self-completion questionnaire distributed in a hardcopy form to the sample. An individual’s level of scepticism and involvement appears to affect the development of a positive effect on employees’ WOM. Involvement with the domain of the investment may be a central factor affecting relationship building within the organization, and upon generation of positive WOM. The chapter offers a conceptual framework to public relations (PR) and corporate communications practitioners, which may enrich their views and understanding of the use and value of CSR for communication strategies and practices. For-profit organisations are major institutions in today’s society. CSR is proffered as presenting advantages for (at macro level) society and (micro level) the organization and its employees. Concepts, such as involvement and scepticism, which have not been rigorously examined in PR and corporate communication literature, are addressed. By examining employee perceptions, managers and academic researchers gain insights into the acceptance, appreciation and effectiveness of CSR policies and activities upon the employee stakeholder group. This will affect current and future CSR communication strategies. The knowledge acquired from this chapter may be transferable outside the for-profit sector

T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones

Isolation of peptide-specific T-cell clones is highly desirable for determining the role of T-cells in human disease, as well as for the development of therapies and diagnostics. However, generation of monoclonal T-cells with the required specificity is challenging and time-consuming. Here we describe a library-based strategy for the simple parallel detection and isolation of multiple peptide-specific human T-cell clones from CD8+ or CD4+ polyclonal T-cell populations. T-cells were first amplified by CD3/CD28 microbeads in a 96U-well library format, prior to screening for desired peptide recognition. T-cells from peptide-reactive wells were then subjected to cytokine-mediated enrichment followed by single-cell cloning, with the entire process from sample to validated clone taking as little as 6 weeks. Overall, T-cell libraries represent an efficient and relatively rapid tool for the generation of peptide-specific T-cell clones, with applications shown here in infectious disease (Epstein–Barr virus, influenza A, and Ebola virus), autoimmunity (type 1 diabetes) and cancer

Polyolefin–polar block copolymers from versatile new macromonomers

A new metallocene-based polymerization mechanism is elucidated in which a zirconium hydride center inserts α-methylstyrene at the start of a polymer chain. The hydride is then regenerated by hydrogenation to release a polyolefin containing a single terminal α-methylstyrenyl group. Through the use of the difunctional monomer 1,3-diisopropenylbenzene, this catalytic hydride insertion polymerization is applied to the production of linear polyethylene and ethylene–hexene copolymers containing an isopropenylbenzene end group. Conducting simple radical polymerizations in the presence of this new type of macromonomer leads to diblock copolymers containing a polyolefin attached to an acrylate, methacrylate, vinyl ester, or styrenic segments. The new materials are readily available and exhibit interfacial phenomena, including the mediation of the mixing of immiscible polymer blends

Wide versus narrow excision margins for high-risk, primary cutaneous melanomas: long-term follow-up of survival in a randomised trial.

Background The necessary margin of excision for cutaneous melanomas greater than 2 mm in thickness is controversial. At a median follow-up of 5 years, findings from our previously published randomised trial of narrow (1 cm) versus wide (3 cm) excision margins in patients with thick cutaneous melanomas showed that narrow margins were associated with an increased frequency of locoregional relapse, but no significant difference in overall survival was apparent. We now report a long-term survival analysis of that trial.Methods We did a randomised, open-label multicentre trial in 59 hospitals--57 in the UK, one in Poland, and one in South Africa. Patients with one primary localised cutaneous melanoma greater than 2 mm in Breslow thickness on the trunk or limbs (excluding palms or soles) were randomly assigned (1:1) centrally to receive surgery with either a 1 cm or 3 cm excision margin following an initial surgery. The randomisation lists were generated with random permuted blocks and stratified by centre and extent of initial surgery. The endpoints of this analysis were overall survival and melanoma-specific survival. Analyses were done in the intention-to-treat population. This trial was not registered because it predated mandatory trial registration.Findings Between Dec 16, 1992, and May 22, 2001, we randomly assigned 900 patients to surgery with either a 1 cm excision margin (n=453) or a 3 cm excision margin (n=447). At a median follow-up of 8·8 years (106 months [IQR 76-135], 494 patients had died, with 359 of these deaths attributed to melanoma. 194 deaths were attributed to melanoma in the 1 cm group compared with 165 in the 3 cm group (unadjusted hazard ratio [HR] 1·24 [95% CI 1·01-1·53]; p=0·041). Although a higher number of deaths overall occurred in the 1 cm group compared with the 3 cm group (253 vs 241), the difference was not significant (unadjusted HR 1·14 [95% CI 0·96-1·36]; p=0·14). Surgical complications were reported in 35 (8%) patients in the 1 cm excision margin group and 65 (15%) patients in the 3 cm group.Interpretation Our findings suggest that a 1 cm excision margin is inadequate for cutaneous melanoma with Breslow thickness greater than 2 mm on the trunk and limbs. Current guidelines advise a 2 cm margin for melanomas greater than 2 mm in thickness but only a 1 cm margin for thinner melanomas. The adequacy of a 1 cm margin for thinner melanomas with poor prognostic features should be addressed in future randomised studies.Funding Cancer Research UK, North Thames National Health Service Executive, Northern and Yorkshire National Health Service Executive, British United Provident Association Foundation, British Association of Plastic Surgeons, the Meirion Thomas Cancer Research Fund, and the National Institute for Health and Research Biomedical Research Centre at The Royal Marsden NHS Foundation Trust

Knowledge and behavior of Nigerian dentists concerning the treatment of children with special needs

BACKGROUND: Children with special needs (CSN) are reported to receive less adequate dental care due to various behavioral problems and barriers created by dental professionals. This study was carried out to determine the knowledge and behaviour of Nigerian dentists concerning the treatment of CSN. METHODS: Questionnaires consisting of open and closed ended questions requesting socio-demographic information, type of practice, undergraduate and postgraduate training, self-rated knowledge and behaviour concerning care of CSN, were hand delivered to 359 dentists in the 3 geographical zones of Nigeria over a period of 8 weeks. Responses were compared across age groups, gender, type of practice and training received. RESULT: Two hundred and eighty questionnaires were returned completed, constituting 79.9% response rate. Most of the respondents were aged 30 – 39 years (44.3%). There were more males (56.1%) and more recent graduates of 10 years and below (78.5%). Over 80% of respondents had treated children with disabilities, those with physical disabilities being most encountered. Only 19.3% of respondents rated their knowledge of management of CSN as adequate, with no significant difference across age groups and gender, but with a significantly higher number of older graduates reporting to have adequate knowledge (p < 0.05). Those who had undergraduate training in care of CSN were 69.5% compared with only 12.8% who had post graduate training. Only 11.8% rated their undergraduate training as adequate. Thirty seven percent of respondents rated the CSN they had treated as very challenging. A higher proportion of older graduates (of more than 10 years post graduation) and those who rated their undergraduate training as inadequate used sedation and general anaesthesia. Seventy one percent of respondents were willing to treat CSN, with no significant difference across age groups, gender and training, but with a significantly higher percentage among those who had rated their knowledge as adequate. Most of those who were unwilling to treat CSN felt their management was tedious and challenging. CONCLUSION: From this study, very few dentists reported to have adequate knowledge of management of CSN, irrespective of age, gender and place of practice. A significant number of those with more experience rated their knowledge as adequate. Although most dentists rated the children's behaviour as challenging, they indicated their willingness to treat them in their practices