Forbidden fruit (and vegies)

I am interested in macro-photography, particularly in the ways that natural objects or substances, when viewed at close range, are transformed into intriguing abstract patterns or conjure unfamiliar worlds. My favourite subjects for this are bark, sand, rock and snow, and, to a lesser degree, flowers and fruit. The colours, textures and shapes of the everyday things we take for granted or simply pass without noticing can become fascinatingly alive

POT1 mutations predispose to familial melanoma

Deleterious germline variants in CDKN2A account for around 40% of familial melanoma cases, and rare variants in CDK4, BRCA2, BAP1 and the promoter of TERT have also been linked to the disease. Here we set out to identify new high-penetrance susceptibility genes by sequencing 184 melanoma cases from 105 pedigrees recruited in the UK, The Netherlands and Australia that were negative for variants in known predisposition genes. We identified families where melanoma cosegregates with loss-of-function variants in the protection of telomeres 1 gene (POT1), with a proportion of family members presenting with an early age of onset and multiple primary tumors. We show that these variants either affect POT1 mRNA splicing or alter key residues in the highly conserved oligonucleotide/oligosaccharide-binding (OB) domains of POT1, disrupting protein-telomere binding and leading to increased telomere length. These findings suggest that POT1 variants predispose to melanoma formation via a direct effect on telomeres.D.J.A., C.D.R.-E., Z.D., J.Z.L., J.C.T., M.P. and T.M.K. were supported by Cancer Research UK and the Wellcome Trust (WT098051). C.D.R.-E. was also supported by the Consejo Nacional de Ciencia y Tecnología of Mexico. K.A.P. and A.M.D. were supported by Cancer Research UK (grants C1287/A9540 and C8197/A10123) and by the Isaac Newton Trust. N.K.H. was supported by a fellowship from the National Health and Medical Research Council of Australia (NHMRC). L.G.A. was supported by an Australia and New Zealand Banking Group Limited Trustees PhD scholarship. A.L.P. is supported by Cure Cancer Australia. The work was funded in part by the NHMRC and Cancer Council Queensland. The work of N.A.G. was in part supported by the Dutch Cancer Society (UL 2012-5489). M.H., J.A.N.-B. and D.T.B. were supported by Cancer Research UK (programme awards C588/A4994 and C588/A10589 and the Genomics Initiative). C.L.-O., A.J.R. and V.Q. are funded by the Spanish Ministry of Economy and Competitiveness through the Instituto de Salud Carlos III (ISCIII), the Red Temática de Investigación del Cáncer (RTICC) del ISCIII and the Consolider-Ingenio RNAREG Consortium. C.L.-O. is an investigator with the Botín Foundation.This is the author accepted manuscript. The final version is available from NPG via http://dx.doi.org/10.1038/ng.294

Economic Globalization, Nutrition and Health: a review of quantitative evidence

BACKGROUND: Unhealthy dietary patterns have in recent decades contributed to an endemic-level burden from non-communicable disease (NCDs) in high-income countries. In low- and middle-income countries rapid changes in diets are also increasingly linked to malnutrition in all its forms as persistent undernutrition and micronutrient deficiencies continue to coexist with a rising prevalence of obesity and associated NCDs. Economic globalization and trade liberalization have been identified as potentially important factors driving these trends, but the mechanisms, pathways and actual impact are subject to continued debate. METHODS: We use a ‘rigorous review’ to synthesize evidence from empirical quantitative studies analysing the links between economic globalization processes and nutritional outcomes, with a focus on impact as well as improving the understanding of the main underlying mechanisms and their interactions. FINDINGS: While the literature remains mixed regarding the impacts of overall globalization, trade liberalization or economic globalization on nutritional outcomes, it is possible to identify different patterns of association and impact across specific sub-components of globalization processes. Although results depend on the context and methods of analysis, foreign direct investment (FDI) appears to be more clearly associated with increases in overnutrition and NCD prevalence than to changes in undernutrition. Existing evidence does not clearly show associations between trade liberalization and NCD prevalence, but there is some evidence of a broad association with improved dietary quality and reductions in undernutrition. Socio-cultural aspects of globalization appear to play an important yet under-studied role, with potential associations with increased prevalence of overweight and obesity. The limited evidence available also suggests that the association between trade liberalization or globalization and nutritional outcomes might differ substantially across population sub-groups. Overall, our findings suggest that policymakers do not necessarily face a trade-off when considering the implications of trade or economic liberalization for malnutrition in all its forms. On the contrary, a combination of nutrition-sensitive trade policy and adequate regulation of FDI could help reduce all forms of malnutrition. In the context of trade negotiations and agreements it is fundamental, therefore, to protect the policy space for governments to adopt nutrition-sensitive interventions

Inducible but not constitutive expression of PD-L1 in human melanoma cells is dependent on activation of NF-κB.

Monoclonal antibodies against immune checkpoint blockade have proven to be a major success in the treatment of melanoma. The programmed death receptor-1 ligand-1 (PD-L1) expression on melanoma cells is believed to have an inhibitory effect on T cell responses and to be an important escape mechanism from immune attack. Previous studies have shown that PD-L1 can be expressed constitutively or can be induced by IFN-γ secreted by infiltrating lymphocytes. In the present study we have investigated the mechanism underlying these two modes of PD-L1 expression in melanoma cells including cells that had acquired resistance to the BRAF inhibitor vemurafenib. PD-L1 expression was examined by flow cytometry and immunoblotting. Specific inhibitors and siRNA knockdown approaches were used to examine the roles of the RAF/ MEK, PI3K, NF-κB, STAT3 and AP1/ c-Jun pathways. IFN-γ inducible expression of PD-L1 was dependent on NF-κB as shown by inhibition with BMS-345541, an inhibitor of IκB and the BET protein inhibitor I-BET151, as well as by siRNA knockdown of NF-κB subunits. We were unable to implicate the BRAF/MEK pathway as major regulators in PD-L1 expression on vemurafenib resistant cells. Similarly the PI3K/AKT pathway and the transcription factors STAT3 and c-Jun had only minor roles in IFN-γ induced expression of PD-L1. The mechanism underlying constitutive expression remains unresolved. We suggest these results have significance in selection of treatments that can be used in combination with monoclonal antibodies against PD1, to enhance their effectiveness and to reduce inhibitory effects melanoma cells have against cytotoxic T cell activity

Understanding the civic impact of journalism: A realistic evaluation perspective

The importance of journalism to civil society is constantly proclaimed, but empirical evidence on journalism's impact, and how this operates, is surprisingly thin. Indeed, there is confusion even about what is meant by the term “impact”. Meanwhile, the issue of the role of journalism is becoming increasingly urgent as a consequence of the rapid changes engulfing the news media, brought about by technological change and the flow-on effect to the traditional advertising-supported business model. Assessing the impact of journalism has recently been the topic of debate among practitioners and scholars particularly in the United States, where philanthropists have responded to the perceived crisis in investigative journalism by funding not-for-profit newsrooms, with resulting new pressures being placed on journalists and editors to quantify their impact on society. These recent attempts have so far failed to achieve clarity or a satisfactory conclusion, which is not surprising given the complex web of causation within which journalism operates. In this paper, the authors propose a stratified definition of journalistic impact and function. They propose a methodology for studying impact drawing on realistic evaluation—a theory-based approach developed primarily to assess large social programmes occurring in open systems. The authors argue this could allow a conceptual and methodological advance on the question of media impacts, leading to research capable of usefully informing responses at a time of worrying change

Nicotinamide Inhibits T Cell Exhaustion and Increases Differentiation of CD8 Effector T Cells

One of the limitations of immunotherapy is the development of a state referred to as T cell exhaustion (TEx) whereby T cells express inhibitory receptors (IRs) and lose production of effectors involved in killing of their targets. In the present studies we have used the repeated stimulation model with anti CD3 and anti CD28 to understand the factors involved in TEx development and treatments that may reduce changes of TEx. The results show that addition of nicotinamide (NAM) involved in energy supply to cells prevented the development of inhibitory receptors (IRs). This was particularly evident for the IRs CD39, TIM3, and to a lesser extent LAG3 and PD1 expression. NAM also prevented the inhibition of IL-2 and TNFα expression in TEx and induced differentiation of CD4+ and CD8 T cells to effector memory and terminal effector T cells. The present results showed that effects of NAM were linked to regulation of reactive oxygen species (ROS) consistent with previous studies implicating ROS in upregulation of TOX transcription factors that induce TEx. These effects of NAM in reducing changes of TEx and in increasing the differentiation of T cells to effector states appears to have important implications for the use of NAM supplements in immunotherapy against cancers and viral infections and require further exploration in vivo

Do innate killing mechanisms activated by inflammasomes have a role in treating melanoma?

Melanoma, as for many other cancers, undergoes a selection process during progression that limits many innate and adaptive tumor control mechanisms. Immunotherapy with immune checkpoint blockade overcomes one of the escape mechanisms but if the tumor is not eliminated other escape mechanisms evolve that require new approaches for tumor control. Some of the innate mechanisms that have evolved against infections with microorganisms and viruses are proving to be active against cancer cells but require better understanding of how they are activated and what inhibitory mechanisms may need to be targeted. This is particularly so for inflammasomes which have evolved against many different organisms and which recruit a number of cytotoxic mechanisms that remain poorly understood. Equally important is understanding of where these mechanisms will fit into existing treatment strategies and whether existing strategies already involve the innate killing mechanisms

Targeting NF-κB reduces PD-L1.

<p><b>A</b>. KMJR138 cells were treated with DMSO (control) or 100ng/ml IFN-γ (IFN) or indicated doses of BMS-345541 in the presence of IFN-γ for 48 hours before PD-L1 was analysed by flow cytometry. Mean Fluorescence Intensity (MFI) of PD-L1 expression is indicated; the average of two independent replicates is shown. <b>B</b>. Indicated cells were treated with either dimethyl sulfoxide (DMSO) (control) or 10μM of I-BET151 (IBET) or 5μM of BMS-345541 (BMS) for 48 hours in the absence or presence of 100ng/ml IFN-γ and PD-L1 cell surface expression was determined by flow cytometry. Mean fluorescence intensity (MFI) plotted is the average of three independent experiments. <b>C</b>. Sub-cellular fractionation of cellular lysates from a representative cell line (KMJR138) was probed for the indicated proteins. Cells were treated with DMSO (control), IFN-γ (IFN, +), I-BET151 (IBET), BMS-345541 (BMS) for 48 hours and separated into cytosolic (C) or nuclear (N) fractions. One representative blot out of two independent experiments is shown. <b>D</b>. KMJR138 cells treated with DMSO (control) or 100ng/ml IFN-γ, 5μM BMS-345541 or a combination of both for 48 hours in the absence or presence of 10μM pan-caspase inhibitor Q-VD-OPh were stained with Annexin V-APC, and PI to determine the number of apoptotic cells (left panel) and anti-PD-L1 antibody to determine the expression of PD-L1 (right panel) which is represented as the mean fluorescence intensity (MFI) over isotype controls. Average value from three independent experiments is indicated. <b>E.</b> Dual renilla-luciferase NF-κB reporter assay: KMJR138 cells were transfected with the control and NF-κB constructs for 24 hours and treated with the indicated inhibitors (10μM I-BET151 or 5μM BMS-345541) with or without 100ng/ml IFN-γ. Luciferase was measured and normalised to renilla values to determine promoter activity. Transfected cells were treated with the inhibitors for 24 hours and IFN-γ induction was carried out 1 hour prior to harvesting and recording luminescence. ***p-value = 0.0001; **p-value = 0.001; *p-value = 0.01. Average of two independent replicates experiments is indicated</p

Effect of signaling pathway inhibition in cells expressing high levels of PD-L1.

<p><b>A</b>. Patient 1 pre cells were treated with DMSO (control) or BRAF inhibitors (dabrafenib 100nM/ml or vemurafenib 10μM/ml) or 10μM of MEK inhibitor UO126 or 40μM PI3K inhibitor LY29004 for 48 hours and the average PD-L1 expression determined by flow cytometry is represented as a fold difference of the mean fluorescence intensity compared to isotype levels. The plotted average mean fluorescence intensity and SEM are derived from three independent experiments. Western blots for indicated proteins after treatment for 24 hours with trametinib or cobemitinib, LY290024, BKM120 and BEZ235 are also shown. Lanes were from the same blot and a representative blot out of two independent experiments is shown. Patient 1 pre cells were further treated with 10μM I-BET151 (IBET) or 5μM BMS-345541 (BMS) for 48 hours and PD-L1 expression determined by western blotting. <b>B.</b> Patient 1 pre cells were transfected with siRNA against the p65 and p50 subunits of NF-κB or control silencer (-) for 72 hours. Cellular lysates were immunoblotted for the indicated proteins. One representative blot out of three independent experiments is indicated. <b>C</b>. Cells were transduced using lentiviral constructs of either control pSIH-copGFP or STAT3-copGFP for 72 hours. Immunoblot for the indicated proteins is shown. Cells were also independently transfected with the control or SMART Pool c-Jun silencer for 48 hours and the cellular lysates were immunoblotted for the indicated proteins. One representative blot out of two independent experiments is indicated.</p

NF-κB is required for PD-L1 expression.

<p><b>A</b>. Indicated cells were transfected with control si (-) or siRNAs against p65 (+) for 48 hours and treated with 100ng/ml IFN-γ for an additional 48 hours before they were harvested for immunoblotting for the indicated proteins. V2 si RNA transfection was done independently with the accompanying control siRNA. One representative out of three independent blots is shown. <b>B</b>. <b>C</b>.KMJR138 cells were transfected with either control (-) or two independent siRNA targeting p105/p50 (+) for 48 hours and treated with 100ng/ml IFN-γ for an additional 48 hours before being harvested for immunoblotting for the indicated proteins. One representative out of three independent blots is shown. <b>C.</b> Cells were transfected with either IκBwt-GFP (control vector) or IκBmutant-GFP (super-repressor) using lipofectamine 2000 for 48 hours after which 100ng/ml IFN-γ was added. PD-L1 expression was detected by flow cytometry on GFP gated cells and is represented as MFI; the average of two independent experiments is indicated. *p-value = 0.01.</p