Which colour do you want for your cells, pink or blue? -Improving the synthesis of cyanine dyes

Cyanine dyes are fluorescent compounds commonly used in biosensors because of their great compatibility in vivo and high extinction coefficient, which has made it the perfect choice for our previous studies investigations into prostate cancer. Although cyanine dyes are commercially available, they are very expensive with only few suppliers and problematic synthesis. To facilitate our cancer research projects, we have sought to develop improved synthetic routes to these useful compounds. In this presentation I will describe our work towards the improved synthesis of some of the cyanine dyes, Cy3 and Cy5. The difficulties (and successes) of the synthesis will be analysed and extension of these methods to other members of the family will be discussed

Synthesis of Selective CDK2/SPY1 Inhibitors employing Stereochemical Control - An invaluable tool in an Organic Chemist’s belt

The cell cycle of a healthy eukaryotic cell depends on the efficiency of cyclin-dependent kinase (CDKs) checkmarks, to ensure normal cell proliferation. CDK2 is responsible for progression of cells into the S and M phases, and it is critical to the abnormal growth processes of cancer cells. Examination of different kinds of human cancers, for their vulnerability to CDK2 inhibition, has revealed CDK2 as a good therapeutic target. In the past two decades, various CDK2 inhibitors have been designed but have stumbled on the roadblock of selectivity issue, since CDK2 shares 74 and 68% sequence identity and active sites with its family members’ CDK3 and CDK1, respectively. Moreover, it’s not CDK2 alone that needs to be targeted but the activated complex it forms with Spy1, a protein that can activate CDK2 in the same way as cyclin but is highly upregulated in cancer cells. After extensive computational studies, we found some unique yet challenging CDK2/SPY1 inhibitors. In this presentation, I will discuss the importance of stereochemical control in the design and synthesis of novel and selective CDK2/SPY1 inhibitors. The synthesis ensures that the inhibitors are stereochemically pure, and thus the biological activity can be accurately evaluated. These results can then be used to refine our computational models to further improve the selectivity of our drug candidates

Drug Discovery: Towards the Synthesis of Novel CDK2-Spy1 Inhibitors

As vital regulatory proteins in the cell cycle, cyclin-dependent kinases (CDKs) and cyclins ensure normal cell division and growth by monitoring check points in the cell cycle. CDKs are inactive on its own, but when a cyclin binds to CDK the activated CDK-Cyclin complexes then carry out their role as cell cycle regulators. Unregulated CDK-Cyclin complexes cause cells to grow and divide at a premature stage, and that can lead to uncontrolled cell growth. Existing treatments such as CKI (cyclin-dependent kinase inhibitor) therapy have cytotoxicity issues because of their inability to differentiate cancer cells from healthy cells, resulting in unwanted side effects. Our collaborators in the Porter Lab have identified a new target: CDK2-Spy1 complex. Spy proteins are alternative activators to CDKs in cancer cells, but not in healthy cells, making them an ideal therapeutic target. Notably, the Spy1 gene is among the top 50 genes associated with carcinoma, yet the CDK2-Spy1 complex has never been selectively targeted before in terms of CKI therapy. We therefore aim to synthesize molecules that selectively target CDK2-Spy1 complexes to develop a chemotherapy that has minimal cytotoxicity issues. In this presentation I will discuss our current progress towards small molecule inhibitors that show promising selectivity for CDK2-Spy1 complexes based on computational studies. Our synthetic routes and the analytical techniques employed to characterise these compounds will be described

Understanding Lived Experiences of Stigma for People Living with HIV: A Community Based Participatory Research Study

The goal of this project was to better understand the experiences and impacts of HIV stigma and discrimination on people living with HIV and to co-create knowledge that has the potential to challenge existing stigma within the healthcare, social services, and public policy sectors in the province of Alberta, Canada. We employed community-based participatory research and a mixed methods design (survey methods and qualitative interviews) to address these questions. An online survey was completed by 148 people living with HIV and semi-structured interviews were conducted with an additional 20 participants. The research findings have been conceptualized within a social ecological model. The four main categories that emerged from the data included personal level factors attributed to HIV stigma, interpersonal factors related to HIV stigma, community factors related to HIV stigma, and HIV stigma in systems and institutions. Within each ecological domain we highlight the strengths and coping strategies people living with HIV identified in the study. Results will be of interest to health researchers and HIV service providers

New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk.

Levels of circulating glucose are tightly regulated. To identify new loci influencing glycemic traits, we performed meta-analyses of 21 genome-wide association studies informative for fasting glucose, fasting insulin and indices of beta-cell function (HOMA-B) and insulin resistance (HOMA-IR) in up to 46,186 nondiabetic participants. Follow-up of 25 loci in up to 76,558 additional subjects identified 16 loci associated with fasting glucose and HOMA-B and two loci associated with fasting insulin and HOMA-IR. These include nine loci newly associated with fasting glucose (in or near ADCY5, MADD, ADRA2A, CRY2, FADS1, GLIS3, SLC2A2, PROX1 and C2CD4B) and one influencing fasting insulin and HOMA-IR (near IGF1). We also demonstrated association of ADCY5, PROX1, GCK, GCKR and DGKB-TMEM195 with type 2 diabetes. Within these loci, likely biological candidate genes influence signal transduction, cell proliferation, development, glucose-sensing and circadian regulation. Our results demonstrate that genetic studies of glycemic traits can identify type 2 diabetes risk loci, as well as loci containing gene variants that are associated with a modest elevation in glucose levels but are not associated with overt diabetes

Effective health care for older people living and dying in care homes: A realist review

Background: Care home residents in England have variable access to health care services. There is currently no coherent policy or consensus about the best arrangements to meet these needs. The purpose of this review was to explore the evidence for how different service delivery models for care home residents support and/or improve wellbeing and health-related outcomes in older people living and dying in care homes. Methods: We conceptualised models of health care provision to care homes as complex interventions. We used a realist review approach to develop a preliminary understanding of what supported good health care provision to care homes. We completed a scoping of the literature and interviewed National Health Service and Local Authority commissioners, providers of services to care homes, representatives from the Regulator, care home managers, residents and their families. We used these data to develop theoretical propositions to be tested in the literature to explain why an intervention may be effective in some situations and not others. We searched electronic databases and related grey literature. Finally the findings were reviewed with an external advisory group. Results: Strategies that support and sustain relational working between care home staff and visiting health care professionals explained the observed differences in how health care interventions were accepted and embedded into care home practice. Actions that encouraged visiting health care professionals and care home staff jointly to identify, plan and implement care home appropriate protocols for care, when supported by ongoing facilitation from visiting clinicians, were important. Contextual factors such as financial incentives or sanctions, agreed protocols, clinical expertise and structured approaches to assessment and care planning could support relational working to occur, but of themselves appeared insufficient to achieve change. Conclusion: How relational working is structured between health and care home staff is key to whether health service interventions achieve health related outcomes for residents and their respective organisations. The belief that either paying clinicians to do more in care homes and/or investing in training of care home staff is sufficient for better outcomes was not supported.This research was funded by National Institute of Health Research Health Service Delivery and Research programme (HSDR 11/021/02)

Long-term species, sexual and individual variations in foraging strategies of fur seals revealed by stable isotopes in whiskers

Background: Individual variations in the use of the species niche are an important component of diversity in trophic interactions. A challenge in testing consistency of individual foraging strategy is the repeated collection of information on the same individuals. Methodology/Principal Findings: The foraging strategies of sympatric fur seals (Arctocephalus gazella and A. tropicalis) were examined using the stable isotope signature of serially sampled whiskers. Most whiskers exhibited synchronous delta C-13 and delta N-15 oscillations that correspond to the seal annual movements over the long term (up to 8 years). delta C-13 and delta N-15 values were spread over large ranges, with differences between species, sexes and individuals. The main segregating mechanism operates at the spatial scale. Most seals favored foraging in subantarctic waters (where the Crozet Islands are located) where they fed on myctophids. However, A. gazella dispersed in the Antarctic Zone and A. tropicalis more in the subtropics. Gender differences in annual time budget shape the seal movements. Males that do not perform any parental care exhibited large isotopic oscillations reflecting broad annual migrations, while isotopic values of females confined to a limited foraging range during lactation exhibited smaller changes. Limited inter-individual isotopic variations occurred in female seals and in male A. tropicalis. In contrast, male A. gazella showed large inter-individual variations, with some males migrating repeatedly to high-Antarctic waters where they fed on krill, thus meaning that individual specialization occurred over years. Conclusions/Significance: Whisker isotopic signature yields unique long-term information on individual behaviour that integrates the spatial, trophic and temporal dimensions of the ecological niche. The method allows depicting the entire realized niche of the species, including some of its less well-known components such as age-, sex-, individual- and migration-related changes. It highlights intrapopulation heterogeneity in foraging strategies that could have important implications for likely demographic responses to environmental variability

Pre-Bilaterian Origins of the Hox Cluster and the Hox Code: Evidence from the Sea Anemone, Nematostella vectensis

BACKGROUND: Hox genes were critical to many morphological innovations of bilaterian animals. However, early Hox evolution remains obscure. Phylogenetic, developmental, and genomic analyses on the cnidarian sea anemone Nematostella vectensis challenge recent claims that the Hox code is a bilaterian invention and that no “true” Hox genes exist in the phylum Cnidaria. METHODOLOGY/PRINCIPAL FINDINGS: Phylogenetic analyses of 18 Hox-related genes from Nematostella identify putative Hox1, Hox2, and Hox9+ genes. Statistical comparisons among competing hypotheses bolster these findings, including an explicit consideration of the gene losses implied by alternate topologies. In situ hybridization studies of 20 Hox-related genes reveal that multiple Hox genes are expressed in distinct regions along the primary body axis, supporting the existence of a pre-bilaterian Hox code. Additionally, several Hox genes are expressed in nested domains along the secondary body axis, suggesting a role in “dorsoventral” patterning. CONCLUSIONS/SIGNIFICANCE: A cluster of anterior and posterior Hox genes, as well as ParaHox cluster of genes evolved prior to the cnidarian-bilaterian split. There is evidence to suggest that these clusters were formed from a series of tandem gene duplication events and played a role in patterning both the primary and secondary body axes in a bilaterally symmetrical common ancestor. Cnidarians and bilaterians shared a common ancestor some 570 to 700 million years ago, and as such, are derived from a common body plan. Our work reveals several conserved genetic components that are found in both of these diverse lineages. This finding is consistent with the hypothesis that a set of developmental rules established in the common ancestor of cnidarians and bilaterians is still at work today

A systematic review of rodent pest research in Afro-Malagasy small-holder farming systems: Are we asking the right questions?

Rodent pests are especially problematic in terms of agriculture and public health since they can inflict considerable economic damage associated with their abundance, diversity, generalist feeding habits and high reproductive rates. To quantify rodent pest impacts and identify trends in rodent pest research impacting on small-holder agriculture in the Afro-Malagasy region we did a systematic review of research outputs from 1910 to 2015, by developing an a priori defined set of criteria to allow for replication of the review process. We followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. We reviewed 162 publications, and while rodent pest research was spatially distributed across Africa (32 countries, including Madagascar), there was a disparity in number of studies per country with research biased towards four countries (Tanzania [25%], Nigeria [9%], Ethiopia [9%], Kenya [8%]) accounting for 51% of all rodent pest research in the Afro-Malagasy region. There was a disparity in the research themes addressed by Tanzanian publications compared to publications from the rest of the Afro-Malagasy region where research in Tanzania had a much more applied focus (50%) compared to a more basic research approach (92%) in the rest of the Afro-Malagasy region. We found that pest rodents have a significant negative effect on the Afro-Malagasy small-holder farming communities. Crop losses varied between cropping stages, storage and crops and the highest losses occurred during early cropping stages (46% median loss during seedling stage) and the mature stage (15% median loss). There was a scarcity of studies investigating the effectiveness of various management actions on rodent pest damage and population abundance. Our analysis highlights that there are inadequate empirical studies focused on developing sustainable control methods for rodent pests and rodent pests in the Africa-Malagasy context is generally ignored as a research topic

Genome-wide association analysis identifies six new loci associated with forced vital capacity

Forced vital capacity (FVC), a spirometric measure of pulmonary function, reflects lung volume and is used to diagnose and monitor lung diseases. We performed genome-wide association study meta-analysis of FVC in 52,253 individuals from 26 studies and followed up the top associations in 32,917 additional individuals of European ancestry. We found six new regions associated at genome-wide significance (P < 5 × 10−8) with FVC in or near EFEMP1, BMP6, MIR129-2–HSD17B12, PRDM11, WWOX and KCNJ2. Two loci previously associated with spirometric measures (GSTCD and PTCH1) were related to FVC. Newly implicated regions were followed up in samples from African-American, Korean, Chinese and Hispanic individuals. We detected transcripts for all six newly implicated genes in human lung tissue. The new loci may inform mechanisms involved in lung development and the pathogenesis of restrictive lung disease