Current status of pharmacogenomics testing for anti-tumor drug therapies : approaches to non-melanoma skin cancer

Skin cancer is one of the most commonly occurring cancer types, with substantial social, physical, and financial burdens on both individuals and societies. Although the role of UV light in initiating skin cancer development has been well characterized, genetic studies continue to show that predisposing factors can influence an individual's susceptibility to skin cancer and response to treatment. In the future, it is hoped that genetic profiles, comprising a number of genetic markers collectively involved in skin cancer susceptibility and response to treatment or prognosis, will aid in more accurately informing practitioners' choices of treatment. Individualized treatment based on these profiles has the potential to increase the efficacy of treatments, saving both time and money for the patient by avoiding the need for extensive or repeated treatment. Increased treatment responses may in turn prevent recurrence of skin cancers, reducing the burden of this disease on society. Currently existing pharmacogenomic tests, such as those that assess variation in the metabolism of the anticancer drug fluorouracil, have the potential to reduce the toxic effects of anti-tumor drugs used in the treatment of non-melanoma skin cancer (NMSC) by determining individualized appropriate dosage. If the savings generated by reducing adverse events negate the costs of developing these tests, pharmacogenomic testing may increasingly inform personalized NMSC treatment.

Droplet digital PCR as a useful tool for the quantitative detection of Enterovirus 71

Hand, foot and mouth disease (HFMD) is a contagious viral disease that frequently affects infants and children and present with blisters and flu-like symptoms. This disease is caused by a group of enteroviruses such as enterovirus 71 (EV71) and coxsackievirus A16 (CA16). However, unlike other HFMD causing enteroviruses, EV71 have also been shown to be associated with more severe clinical manifestation such as aseptic meningitis, brainstem and cerebellar encephalitis which may lead to cardiopulmonary failure and death. Clinically, HFMD caused by EV71 is indistinguishable from other HFMD causing enteroviruses such as CA16. Molecular diagnosis methods such as the use of real-time PCR has been used commonly for the identification of EV71. In this study, two platforms namely the real-time PCR and the droplet digital PCR were compared for the detection quantitation of known EV71 viral copy number. The results reveal accurate and consistent results between the two platforms. In summary, the droplet digital PCR was demonstrated to be a promising technology for the identification and quantitation of EV71 viral copy number.

IASDR 2015 Interplay Exhibition 2-5 NOV 2015

It has been said that we are living in a golden age of innovation. New products, systems and services aimed to enable a better future, have emerged from novel interconnections between design and design research with science, technology and the arts. These intersections are now, more than ever, catalysts that enrich daily activities for health and safety, education, personal computing, entertainment and sustainability, to name a few. Interactive functions made possible by new materials, technology, and emerging manufacturing solutions demonstrate an ongoing interplay between cross-disciplinary knowledge and research. Such interactive interplay bring up questions concerning: (i) how art and design provide a focus for developing design solutions and research in technology; (ii) how theories emerging from the interactions of cross-disciplinary knowledge inform both the practice and research of design and (iii) how research and design work together in a mutually beneficial way. The IASDR2015 INTERPLAY EXHIBITION provides some examples of these interconnections of design research with science, technology and the arts. This is done through the presentation of objects, artefacts and demonstrations that are contextualised into everyday activities across various areas including health, education, safety, furniture, fashion and wearable design. The exhibits provide a setting to explore the various ways in which design research interacts across discipline knowledge and approaches to stimulate innovation. In education, Designing South African Children’s Health Education as Generative Play (A Bennett, F Cassim, M van der Merwe, K van Zijil, and M Ribbens) presents a set of toolkits that resulted from design research entailing generative play. The toolkits are systems that engender pleasure and responsibility, and are aimed at cultivating South African’s youth awareness of nutrition, hygiene, disease awareness and prevention, and social health. In safety, AVAnav: Avalanche Rescue Helmet (Jason Germany) delivers an interactive system as a tool to contribute to reduce the time to locate buried avalanche victims. Helmet-mounted this system responds to the contextual needs of rescuers and has since led to further design research on the interface design of rescuing devices. In apparel design and manufacturing, Shrinking Violets: Fashion design for disassembly (Alice Payne) proposes a design for disassembly through the use of beautiful reversible mono-material garments that interactively responds to the challenges of garment construction in the fashion industry, capturing the metaphor for the interplay between technology and craft in the fashion manufacturing industry. Harvest: A biotextile future (Dean Brough and Alice Payne), explores the interplay of biotechnology, materiality and textile design in the creation of sustainable, biodegradable vegan textile through the process of a symbiotic culture of bacteria and yeast (SCOBY). SCOBY is a pellicle curd that can be harvested, machine washed, dried and cut into a variety of designs and texture combinations. The exploration of smart materials, wearable design and micro-electronics led to creative and aesthetically coherent stimulus-reactive jewellery; Symbiotic Microcosms: Crafting Digital Interaction (K Vones). This creation aims to bridge the gap between craft practitioner and scientific discovery, proposing a move towards the notion of a post-human body, where wearable design is seen as potential ground for new human-computer interactions, affording the development of visually engaging multifunctional enhancements. In furniture design, Smart Assistive chair for older adults (Chao Zhao) demonstrates how cross-disciplinary knowledge interacting with design strategies provide solution that employed new technological developments in older aged care, and the participation of multiple stakeholders: designers, health care system and community based health systems. In health, Molecular diagnosis system for newborns deafness genetic screening (Chao Zhao) presents an ambitious and complex project that includes a medical device aimed at resolving a number of challenges: technical feasibility for city and rural contexts, compatibility with standard laboratory and hospital systems, access to health system, and support the work of different hospital specialists. The interplay between cross-disciplines is evident in this work, demonstrating how design research moves forward through technology developments. These works exemplify the intersection between domains as a means to innovation. Novel design problems are identified as design intersects with the various areas. Research informs this process, and in different ways. We see the background investigation into the contextualising domain (e.g. on-snow studies, garment recycling, South African health concerns, the post human body) to identify gaps in the area and design criteria; the technologies and materials reviews (e.g. AR, biotextiles) to offer plausible technical means to solve these, as well as design criteria. Theoretical reviews can also inform the design (e.g. play, flow). These work together to equip the design practitioner with a robust set of ‘tools’ for design innovation – tools that are based in research. The process identifies innovative opportunity and criteria for design and this, in turn, provides a means for evaluating the success of the design outcomes. Such an approach has the potential to come full circle between research and design – where the design can function as an exemplar, evidencing how the research-articulated problems can be solved. Core to this, however, is the evaluation of the design outcome itself and identifying knowledge outcomes. In some cases, this is fairly straightforward that is, easily measurable. For example the efficacy of Jason Germany’s helmet can be determined by measuring the reduced response time in the rescuer. Similarly the improved ability to recycle Payne’s panel garments can be clearly determined by comparing it to those recycling processes (and her identified criteria of separating textile elements!); while the sustainability and durability of the Brough & Payne’s biotextile can be assessed by documenting the growth and decay processes, or comparative strength studies. There are however situations where knowledge outcomes and insights are not so easily determined. Many of the works here are open-ended in their nature, as they emphasise the holistic experience of one or more designs, in context: “the end result of the art activity that provides the health benefit or outcome but rather, the value lies in the delivery and experience of the activity” (Bennet et al.) Similarly, reconfiguring layers of laser cut silk in Payne’s Shrinking Violets constitutes a customisable, creative process of clothing oneself since it “could be layered to create multiple visual effects”. Symbiotic Microcosms also has room for facilitating experience, as the work is described to facilitate “serendipitous discovery”. These examples show the diverse emphasis of enquiry as on the experience versus the product. Open-ended experiences are ambiguous, multifaceted and differ from person to person and moment to moment (Eco 1962). Determining the success is not always clear or immediately discernible; it may also not be the most useful question to ask. Rather, research that seeks to understand the nature of the experience afforded by the artefact is most useful in these situations. It can inform the design practitioner by helping them with subsequent re-design as well as potentially being generalizable to other designers and design contexts. Bennett et. al exemplify how this may be approached from a theoretical perspective. This work is concerned with facilitating engaging experiences to educate and, ultimately impact on that community. The research is concerned with the nature of that experience as well, and in order to do so the authors have employed theoretical lenses – here these are of flow, pleasure, play. An alternative or complementary approach to using theory, is using qualitative studies such as interviews with users to ask them about what they experienced? Here the user insights become evidence for generalising across, potentially revealing insight into relevant concerns – such as the range of possible ‘playful’ or experiences that may be afforded, or the situation that preceded a ‘serendipitous discovery’. As shown, IASDR2015 INTERPLAY EXHIBITION provides a platform for exploration, discussion and interrogation around the interplay of design research across diverse domains. We look forward with excitement as IASDR continues to bring research and design together, and as our communities of practitioners continue to push the envelope of what is design and how this can be expanded and better understood with research to foster new work and ultimately, stimulate innovation.

Molecular approaches to diagnosis and management of ovarian cancer

The recent advances in the understanding of the pathogenesis of ovarian cancer have been helpful in addressing issues in diagnosis, prognosis and management. The study of ovarian tumours by novel techniques such as immunohistochemistry, fluorescent in situ hybridisation, comparative genomic hybridisation, polymerase chain reaction and new tumour markers have aided the evaluation and application of new concepts into clinical practice. The correlation of novel surrogate tumour specific features with response to treatment and outcome in patients has defined prognostic factors which may allow the future design of tailored therapy based on a molecular profile of the tumour. These have also been used to design new approaches to therapy such as antibody targeting and gene therapy. The delineation of roles of c-erbB2, c-fms and other novel receptor kinases in the pathogenesis of ovarian cancer has led initially to the development of anti-c-erbB2 monoclonal antibody therapy. The discovery of BRCA1 and BRCA2 genes will have an impact in the diagnosis and the prevention of familial ovarian cancer. The important role played by recessive genes such as p53 in cancer has raised the possibility of restoration of gene function by gene therapy. Although the pathological diagnosis of ovarian cancer is still confirmed principally on morphological features, addition of newer investigations will increasingly be useful in addressing difficult diagnostic problems. The increasingly rapid pace of discovery of genes important in disease, makes it imperative that the evaluation of their contribution in the pathogenesis of ovarian cancer is undertaken swiftly, thus improving the overall management of patients and their outcome.

Purine Nucleoside Phosphorylase mediated molecular chemotherapy and conventional chemotherapy: A tangible union against chemoresistant cancer

Background Late stage Ovarian Cancer is essentially incurable primarily due to late diagnosis and its inherent heterogeneity. Single agent treatments are inadequate and generally lead to severe side effects at therapeutic doses. It is crucial to develop clinically relevant novel combination regimens involving synergistic modalities that target a wider repertoire of cells and lead to lowered individual doses. Stemming from this premise, this is the first report of two- and three-way synergies between Adenovirus-mediated Purine Nucleoside Phosphorylase based gene directed enzyme prodrug therapy (PNP-GDEPT), docetaxel and/or carboplatin in multidrug-resistant ovarian cancer cells. Methods The effects of PNP-GDEPT on different cellular processes were determined using Shotgun Proteomics analyses. The in vitro cell growth inhibition in differentially treated drug resistant human ovarian cancer cell lines was established using a cell-viability assay. The extent of synergy, additivity, or antagonism between treatments was evaluated using CalcuSyn statistical analyses. The involvement of apoptosis and implicated proteins in effects of different treatments was established using flow cytometry based detection of M30 (an early marker of apoptosis), cell cycle analyses and finally western blot based analyses. Results Efficacy of the trimodal treatment was significantly greater than that achieved with bimodal- or individual treatments with potential for 10-50 fold dose reduction compared to that required for individual treatments. Of note was the marked enhancement in apoptosis that specifically accompanied the combinations that included PNP-GDEPT and accordingly correlated with a shift in the expression of anti- and pro-apoptotic proteins. PNP-GDEPT mediated enhancement of apoptosis was reinforced by cell cycle analyses. Proteomic analyses of PNP-GDEPT treated cells indicated a dowregulation of proteins involved in oncogenesis or cancer drug resistance in treated cells with accompanying upregulation of apoptotic- and tumour- suppressor proteins. Conclusion Inclusion of PNP-GDEPT in regular chemotherapy regimens can lead to significant enhancement of the cancer cell susceptibility to the combined treatment. Overall, these data will underpin the development of regimens that can benefit patients with late stage ovarian cancer leading to significantly improved efficacy and increased quality of life.

Molecular mechanisms of migraine

Migraine is a common complex disorder that affects a large portion of the population and thus incurs a substantial economic burden on society. The disorder is characterized by recurrent headaches that are unilateral and usually accompanied by nausea, vomiting, photophobia, and phonophobia. The range of clinical characteristics is broad and there is evidence of comorbidity with other neurological diseases, complicating both the diagnosis and management of the disorder. Although the class of drugs known as the triptans (serotonin 5-HT1B/1D agonists) has been shown to be effective in treating a significant number of patients with migraine, treatment may in the future be further enhanced by identifying drugs that selectively target molecular mechanisms causing susceptibility to the disease. Genetically, migraine is a complex familial disorder in which the severity and susceptibility of individuals is most likely governed by several genes that may be different among families. Identification of the genomic variants involved in genetic predisposition to migraine should facilitate the development of more effective diagnostic and therapeutic applications. Genetic profiling, combined with our knowledge of therapeutic response to drugs, should enable the development of specific, individually-tailored treatment.

Eph receptors and their ligands : promising molecular biomarkers and therapeutic targets in prostate cancer

Although at present, there is a high incidence of prostate cancer, particularly in the Western world, mortality from this disease is declining and occurs primarily only from clinically significant late stage tumors with a poor prognosis. A major current focus of this field is the identification of new biomarkers which can detect earlier, and more effectively, clinically significant tumors from those deemed “low risk”, as well as predict the prognostic course of a particular cancer. This strategy can in turn offer novel avenues for targeted therapies. The large family of Receptor Tyrosine Kinases, the Ephs, and their binding partners, the ephrins, has been implicated in many cancers of epithelial origin through stimulation of oncogenic transformation, tumor angiogenesis, and promotion of increased cell survival, invasion and migration. They also show promise as both biomarkers of diagnostic and prognostic value and as targeted therapies in cancer. This review will briefly discuss the complex roles and biological mechanisms of action of these receptors and ligands and, with regard to prostate cancer, highlight their potential as biomarkers for both diagnosis and prognosis, their application as imaging agents, and current approaches to assessing them as therapeutic targets. This review demonstrates the need for future studies into those particular family members that will prove helpful in understanding the biology and potential as targets for treatment of prostate cancer.

Carcinoma ex pleomorphic adenoma : a comprehensive review of clinical, pathological and molecular data

Carcinoma ex pleomorphic adenoma (Ca ex PA) is a carcinoma arising from a primary or recurrent benign pleomorphic adenoma. It often poses a diagnostic challenge to clinicians and pathologists. This study intends to review the literature and highlight the current clinical and molecular perspectives about this entity. The most common clinical presentation of CA ex PA is of a firm mass in the parotid gland. The proportion of adenoma and carcinoma components determines the macroscopic features of this neoplasm. The entity is difficult to diagnose pre-operatively. Pathologic assessment is the gold standard for making the diagnosis. Treatment for Ca ex PA often involves an ablative surgical procedure which may be followed by radiotherapy. Overall, patients with Ca ex PA have a poor prognosis. Accurate diagnosis and aggressive surgical management of patients presenting with Ca ex PA can increase their survival rates. Molecular studies have revealed that the development of Ca ex PA follows a multi-step model of carcinogenesis, with the progressive loss of heterozygosity at chromosomal arms 8q, then 12q and finally 17p. There are specific candidate genes in these regions that are associated with particular stages in the progression of Ca ex PA. In addition, many genes which regulate tumour suppression, cell cycle control, growth factors and cell-cell adhesion play a role in the development and progression of Ca ex PA. It is hopeful that these molecular data can give clues for the diagnosis and management of the disease.

2nd ESMO consensus conference on lung cancer : early-stage non-small-cell lung cancer consensus on diagnosis, treatment and follow-up

To complement the existing treatment guidelines for all tumour types, ESMO organises consensus conferences to focus on specific issues in each type of tumour. The 2nd ESMO Consensus Conference on Lung Cancer was held on 11-12 May 2013 in Lugano. A total of 35 experts met to address several questions on non-small-cell lung cancer (NSCLC) in each of four areas: pathology and molecular biomarkers, first-line/second and further lines in advanced disease, earlystage disease and locally advanced disease. For each question, recommendations were made including reference to the grade of recommendation and level of evidence. This consensus paper focuses on early-stage disease. © The Author 2014. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved.

Second ESMO consensus conference on lung cancer : pathology and molecular biomarkers for non-small-cell lung cancer

To complement the existing treatment guidelines for all tumour types, ESMO organises consensus conferences to focus on specific issues in each type of tumour. The Second ESMO Consensus Conference on Lung Cancer was held on 11-12 May 2013 in Lugano. A total of 35 experts met to address several questions on management of patients with nonsmall- cell lung cancer (NSCLC) in each of four areas: pathology and molecular biomarkers, early stage disease, locally advanced disease and advanced (metastatic) disease. For each question, recommendations were made including reference to the grade of recommendation and level of evidence. This consensus paper focuses on recommendations for pathology and molecular biomarkers in relation to the diagnosis of lung cancer, primarily non-small-cell carcinomas.

Molecular cloning, expression and immunological characterisation of Pas n 1, the major allergen of Bahia grass Paspalum notatum pollen

Bahia grass, Paspalum notatum, is a clinically important subtropical grass with a prolonged pollination season from spring to autumn. We aimed to clone and characterise the major Bahia grass pollen allergen, Pas n 1. Grass pollen-allergic patients presenting to a tertiary hospital allergy clinic were tested for IgE reactivity with Bahia grass pollen extract by skin prick testing, ImmunoCAP, ELISA and immunoblotting. Using primers deduced from the N-terminal peptide sequence of a group 1 allergen of Bahia grass pollen extract separated by two-dimensional gel electrophoresis, the complete Pas n 1 cDNA was obtained by rapid amplification of cDNA ends and cloned. Biological relevance of recombinant Pas n 1 expressed in Escherichia coli was assessed by serum IgE reactivity and basophil activation. Twenty-nine of 34 (85%) consecutive patients presenting with grass pollen allergy were skin prick test positive to Bahia grass pollen. The Pas n 1 cDNA has sequence homology with the β-expansin 1 glycoprotein family and is more closely related to the maize pollen group 1 allergen (85% identity) than to ryegrass Lol p 1 or Timothy grass Phl p 1 (64 and 66% identity, respectively). rPas n 1 reacted with serum IgE in 47 of 55 (85%) Bahia grass pollen-allergic patients, activated basophils and inhibited serum IgE reactivity with the 29 kDa band of Bahia grass pollen extract. In conclusion the cDNA for the major group 1 allergen of the subtropical Bahia grass pollen, Pas n 1, was identified and cloned. rPas n 1 is immunologically active and is a valuable reagent for diagnosis and specific immunotherapy of grass pollen allergy.

Synthesis of a multimodal molecular imaging probe based on a hyperbranched polymer architecture

Molecular imaging is utilised in modern medicine to aid in the diagnosis and treatment of disease by allowing its spatiotemporal state to be examined in vivo. This study focuses on the development of novel multimodal molecular imaging agents based on hyperbranched polymers that combine the complementary capabilities of optical fluorescence imaging and positron emission tomography-computed tomography (PET/CT) into one construct. RAFT-mediated polymerisation was used to prepare two hydrophilic hyperbranched polymers that were differentiated by their size and level of branching. The multiple functional end-groups facilitated covalent attachment of both near infrared fluorescent dyes for optical imaging, as well as a copper chelator allowing binding of 64Cu as a PET radio nuclei. In vivo multimodal imaging of mice using PET/CT and planar optical imaging was first used to assess the biodistribution of the polymeric materials and it was shown that the larger and more branched polymer had a significantly longer circulation time. The larger constructs were also shown to exhibit enhanced accumulation in solid tumours in a murine B16 melanoma model. Importantly, it was demonstrated that the PET modality gave rise to high sensitivity immediately after injection of the agent, while the optical modality facilitated extended longitudinal studies, thus highlighting how the complementary capabilities of the molecular imaging agents can be useful for studying various diseases, including cancer.