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A set of Quantitative Precipitation Estimates (QPEs) from a dual-polarisation X-band radar observation campaign in a mountainous area of Northern Scotland is assessed with reference to observed river flows as well as being compared to estimates from the UK C-band radar and raingauge networks. Employing estimation methods of varying complexity, the X-band QPEs are trialled as alternative inputs to Grid-to-Grid (G2G), a distributed hydrological model, to produce simulated river flows for comparison with observations. This hydrological assessment complements and extends a previous meteorological assessment that used point raingauge data only. Precipitation estimates for two periods over the observation campaign in 2016 (March to April and June to August) are assessed. During the second period, increased incorporation of dual-polarisation variables into the radar processing chain is found to be of considerable benefit, whereas during the first period the low height of the melting layer often restricts their use. As a result of the complex topography in Northern Scotland, the Lowest Usable Elevation (LUE) of the X-band radar observations is found to be a stronger indicator of the hydrological model performance than range from the radar. For catchments with an LUE of less than 3 km, the best X-band QPE typically performs better for modelling river flow than using an estimate from the UK C-band radar network. The hydrological assessment framework used here brings fresh insights into the performance of the different QPEs, as well as providing a stimulus for targeted improvements to dual-polarisation radar-based QPEs that have wider relevance beyond the case study situation.

The Radar Applications in Northern Scotland (RAiNS) experiment took place from February to August 2016 near Inverness, Scotland. The campaign was motivated by the need to provide enhanced weather radar observations for hydrological applications for the Inverness region. Here we describe the campaign in detail and observations over the summer period of the campaign that show the improvements that high-resolution polarimetric radar observations may have on quantitative precipitation estimates in this region compared to concurrently generated operational radar quantitative precipitation estimates (QPEs). We further provide suggestions of methods for generating QPE using dual-polarization X-band radars in similar regions.

PurposeObesity in children and adolescents is a significant public health concern. The World Health Organization Health Promoting Schools (HPS) framework promotes good nutrition and physical activity in school settings. While HPS is embraced globally, effective implementation and sustainable programmes are a continued challenge. This paper aims to report on the characteristics of current school interventions based on HPS and implementation barriers and enablers.Design/methodology/approachA literature search identified peer-reviewed studies of school health interventions reflective of the HPS framework focusing on obesity prevention. Studies from all countries were included, if conducted in primary and/or secondary schools; included a sufficient amount of qualitative implementation or process evaluation data to draw conclusions regarding key barriers and enablers to implementation; and were published in English.FindingsNine interventions (n = 9) from seven countries were included. Most were implemented in primary schools and focused on specific grade levels. Engaging parents, the home environment, teacher time constraints, fun interventions, student participation, teacher training, integration with the curriculum and stakeholder engagement all emerged as strong implementation themes. Teachers as role models, establishing community partnerships and policy support also emerged as common themes.Originality/valueFuture interventions may benefit from enhancing teacher and parent health promotion. Partnerships with initiatives focusing on environmental sustainability may simultaneously benefit human and planetary health while strengthening stakeholder engagement opportunities and consistent messaging throughout the community. More comprehensive evaluation data are needed, in particular, for long-term HPS initiatives.

Observing, in a quantitative and robust way, the dynamic space-time pattern of precipitation in mountainous terrain presents a major challenge of great practical importance. The difficulties of this task are further exacerbated in mid to high latitudes where the typical melting layer for precipitation (i.e. the 0°C isotherm) is often close to the surface during winter months. One way to address this challenge is by improving observations made using networks of weather radars. Quantitative Precipitation Estimates (QPEs) derived from these instruments have many applications, for example as input to a hydrological model to simulate river flow for flood forecasting purposes. Here, a set of QPEs - obtained from an observation campaign using the National Centre for Atmospheric Science’s mobile X-band dual-polarisation Doppler weather radar (NXPol) in a mountainous area of Northern Scotland - are assessed with reference to observed river flows. Each form of QPE is used as an input to Grid-to-Grid (G2G), a distributed hydrological model used for flood forecasting across Great Britain, and the simulated river flows compared to observations. The location of the radar was specially chosen to infill an area of reduced coverage in the existing C-band radar network for the British Isles.Assessments of radar QPE often only examine a final precipitation “best estimate” product and typically with reference to raingauges at specific locations. Here, we exploit the processing capabilities of NXPol and the hydrological modelling framework to investigate the benefits of ten separate processing methods that increase with complexity and make differing use of dual-polarisation variables. The role of the radar beam elevation and distance from the radar is investigated, and NXPol QPEs are compared to that provided by the radar network. Additionally, a preliminary investigation is carried out into the role of the drop-size distribution on the relationship between radar-reflectivity and rain-rate using disdrometer data.The hydrological assessment reported on here has the benefit of integrating the precipitation over space and time which serves to complement and extend a previous meteorological assessment using raingauge data alone. The assessment proves to be insensitive to issues affecting both raingauges (e.g. representativity, wind-induced under-catch) and local artefacts in the space-time radar-rainfall field. It facilitates a direct assessment of whether potential benefits in the new QPEs are carried forward to an end-use such as flood forecasting, providing fresh insights for the development of new dual-polarisation radar QPE methods.

Background There are limited treatments for triple negative breast cancer (TNBC) patients and an unmet need for targeted approaches in these patients. In the last 4-5 years, the prevalence of high androgen receptor (AR) expression in TNBCs has been noted in up to 50% of tumors suggesting it has clinical relevance. The non-canonical NF-kB pathway is also upregulated in this patient group and it has been reported that that there is crosstalk between these pathways. Therefore the aim of this study was to examine the expression of IKKα and AR in breast cancer tissue samples, to assess if combining these markers increased prognostic power. Methods Immunohistochemistry was performed on tissue microarray of 410 patients to assess proteins level of IKK alpha and AR. Protein expression levels were assessed using the weighted histoscore (WHS) method. The median was employed as the cut off for IKK alpha and 1% as cut off for AR. Expression was analyzed for associations with cancer-specific survival (CSS) and recurrence-free survival (RFS). Results In a cohort of 370 breast cancers nether AR nor IKK alpha alone or combined were associated with CSS or RFS. Stratifying patients by ER status did not impact CSS or RFS. However, in TNBC patients (n=82) high expression of AR was associated with shorter CSS (HR 2.55 95 CI 1.61-5.59, p=0.013). To assess if combining AR and IKK alpha increased prognostic power, AR and IKK alpha were combined into a single score: 0= low expression of both or high expression of one and 1= high expression of both. In the full cohort or when stratified by ER status the score was not associated with CSS or RFS, however in TNBC the combined score potentiated the effect observed with AR alone, (HR 1.68 95 CI 1.20-2.33, p=0.001). Patient CSS was stratified from 11.5 years to 4.6 years and was independently associated with CSS when compared with common clinicopathological factors (HR 1.56 95CI 1.11-2.21, p=0.011). In addition, the combined score was associated with decrease radiotherapy use (p=0.032), increased recurrence rate (p=0.014), decreased cytotoxic T cells (p=0.007), B cells (p=0.043) and macrophages (p=0.037). Conclusions A combined AR and IKK alpha score is an independent prognostic classification for patients with TNBC. Patients with high expression of both AR and IKK alpha a significantly reduced survival and were immune cell cold. This study suggests that this patient group will not benefit from immunotherapy but dual targeting with anti-androgens and IKK alpha selective inhibitors could offer a novel therapeutic strategy for this patient group. Citation Format: Roseweir AK, Khongthong P, Dickson K, Bennett L, Edwards J. Dual targeting of androgen receptor and IKK alpha is a potential therapeutic strategy for triple negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-10-13.

Short tandem repeat polymorphisms (STRs) are the standard markers for forensic human identification. STRs are highly polymorphic loci analyzed using a direct PCR-to-CE (capillary electrophoresis) approach. However, STRs have limitations particularly when dealing with complex mixtures. These include slippage of the polymerase during amplification causing stutter fragments that can be indistinguishable from minor contributor alleles, preferential amplification of shorter alleles, and limited number of loci that can be effectively co-amplified with CE. Massively parallel sequencing (MPS), by enabling a higher level of multiplexing and actual sequencing of the DNA, provides forensic practitioners an increased power of discrimination offered by the sequence of STR alleles and access to new sequence-based markers. Microhaplotypes (i.e., microhaps or MHs) are emerging multi-allelic loci of two or more SNPs within

This paper examines the mechanisms responsible for the production of ice in convective clouds influenced by mineral dust. Observations were made in the Ice in Clouds Experiment – Dust (ICE-D) field campaign which took place in the vicinity of Cape Verde during August 2015. Measurements made with instruments on the FAAM aircraft through the clouds on 21 August showed that ice particles were observed in high concentrations at temperatures greater than about −8 °C. Sensitivity studies were performed using existing parametrisation schemes in a cloud model to explore the impact of the freezing onset temperature, the efficiency of freezing, mineral dust as efficient ice nuclei, and multi-thermals on secondary ice production by the rime-splintering process. The simulation with the default Morrison microphysics scheme (Morrison et al., 2005) that involved a single thermal produced a concentration of secondary ice that was much lower than the observed value of total ice number concentration. Relaxing the onset temperature to a higher value, enhancing the freezing efficiency, or combinations of these, increased the secondary ice particle concentration, but not by a sufficient amount. Simulations that involved only dust particles as ice nucleating particles produced a lower concentration of secondary ice particles, since the freezing onset temperature is low. The simulations implicate that a higher concentration of ice nucleating particles with a higher freezing onset temperature may explain some of the observed high concentrations of secondary ice. However, a simulation with two thermals that used the original Morrison scheme without enhancement or relaxation produced the greatest concentration of secondary ice particles. It did so because of the increased time that graupel particles were exposed to significant cloud liquid water content in the Hallett-Mossop temperature zone. The forward-facing camera and measurements of the vertical wind in repeated passes of the same cloud suggested that these tropical clouds contained multiple thermals. Hence, in a similar way to other convective clouds observed elsewhere in the world, it is likely that multi-thermals play an important role in producing very high concentrations of secondary ice particles in some tropical clouds.

PurposeThe host inflammatory response is an important determinant of cancer outcome, however, the factor/s that regulate this response remains unclear. We aimed to determine if Human Leukocyte Antigen (HLA) class I and tumour cell proliferation are associated with CD8+ infiltration and survival in patients undergoing potentially curative resection for colorectal cancer.MethodsHLA class I expression (W6/32 and B2-microglobulin) and tumour proliferation index (Ki67) were quantified using immunohistochemistry on tissue micro arrays (TMA). The local inflammatory response at the invasive margin was assessed using the Klintrup-Makinen (K-M) score and CD8+ infiltration was assessed at the invasive margin (mCD8+), stroma (sCD8+) and cancer cell nests (cCD8+).ResultsPreserved HLA class I expression was associated with lower Dukes’ stage (p=0.032), lower T stage (p=0.040) and higher cCD8+ (p=0.003). High Ki67 was associated with a good K-M score (p+ (p=0.033), higher sCD8+ (p=0.025) and higher cCD8+ (p+ infiltration within the cancer cell nests. On multivariate survival analysis, preserved HLA class I expression was associated with disease free (HR 0.47 95%CI (0.25–0.89),p=0.020) and cancer specific survival (HR 0.52 95%CI (0.28–0.97),p=0.038).ConclusionThis study suggests that a pronounced local inflammatory response is independently associated with both, HLA class I expression and tumour proliferation.

It is increasingly appreciated that host factors within the tumor center and microenvironment play a key role in dictating colorectal cancer (CRC) outcomes. As a result, the metastatic process has now been defined as a result of epithelial–mesenchymal transition (EMT). Establishment of the role of EMT within the tumor center and its effect on the tumor microenvironment would be beneficial for prognosis and therapeutic intervention in CRC. The present study assessed five immunohistochemical EMT markers within the tumor center on a 185 Stage II/III CRC patient tissue microarray. In 185 patients with CRC, cytoplasmic snail (HR 1.94 95% confidence interval [CI] 1.15–3.29, p = 0.012) and a novel combined EMT score (HR 3.86 95% CI 2.17–6.86, p < 0.001) were associated with decreased cancer‐specific survival. The combined EMT score was also associated with increased tumor budding (p = 0.046), and systemic inflammation (p = 0.007), as well as decreased memory T‐cells within the stroma (p = 0.030) and at the invasive margin (p = 0.035). Furthermore, the combined EMT score was associated with cancer‐specific survival independent of TNM‐stage (HR 4.12 95% CI 2.30–7.39, p < 0.001). In conclusion, a novel combined EMT score stratifies patient's survival in Stage II/III CRC and associates with key factors of tumor metastasis. Therefore, the combined EMT score could be used to identify patients at risk of micrometastases and who may benefit from standard adjuvant therapy, potentially in combination with EMT blockade.

The aim of our study was to examine the relationship between tumour IKKα expression and breast cancer recurrence and survival. Immunohistochemistry was employed in a discovery and a validation tissue microarray to assess the association of tumour IKKα expression and clinico-pathological characteristics. After siRNA-mediated silencing of IKKα, cell viability and apoptosis were assessed in MCF7 and MDA-MB-231 breast cancer cells. In both the discovery and validation cohorts, associations observed between IKKα and clinical outcome measures were potentiated in oestrogen receptor (ER) positive Luminal A tumours. In the discovery cohort, cytoplasmic IKKα was associated with disease-free survival (p = 0.029) and recurrence-free survival on tamoxifen (p

The Convective Precipitation Experiment (COPE) was a joint U.K.–U.S. field campaign held during the summer of 2013 in the southwest peninsula of England, designed to study convective clouds that produce heavy rain leading to flash floods. The clouds form along convergence lines that develop regularly as a result of the topography. Major flash floods have occurred in the past, most famously at Boscastle in 2004. It has been suggested that much of the rain was produced by warm rain processes, similar to some flash floods that have occurred in the United States. The overarching goal of COPE is to improve quantitative convective precipitation forecasting by understanding the interactions of the cloud microphysics and dynamics and thereby to improve numerical weather prediction (NWP) model skill for forecasts of flash floods. Two research aircraft, the University of Wyoming King Air and the U.K. BAe 146, obtained detailed in situ and remote sensing measurements in, around, and below storms on several days. A new fast-scanning X-band dual-polarization Doppler radar made 360° volume scans over 10 elevation angles approximately every 5 min and was augmented by two Met Office C-band radars and the Chilbolton S-band radar. Detailed aerosol measurements were made on the aircraft and on the ground. This paper i) provides an overview of the COPE field campaign and the resulting dataset, ii) presents examples of heavy convective rainfall in clouds containing ice and also in relatively shallow clouds through the warm rain process alone, and iii) explains how COPE data will be used to improve high-resolution NWP models for operational use.

In recent years, mobile dual-polarisation Doppler X-band radars have become a prevalent part of the atmospheric scientist’s toolkit for examining cloud dynamics and microphysics and making quantitative precipitation estimates. Here we describe the National Centre for Atmospheric Science (NCAS) mobile X-band Dual-polarisation Doppler weather radar (NXPol) and the infrastructure used to deploy the radar and provide an overview of the technical specifications. It is the first radar of its kind in the United Kingdom. The NXPol is a Meteor 50DX manufactured by Selex-Gematronik (Selex ES GmbH), modified to operate with a larger 2.4 m diameter antenna that produces a 0.98° half-power beam width and without a radome. We provide an overview of the technical specifications of the NXPol with emphasis given to the description of the aspects of the infrastructure developed to deploy the radar as an autonomous observing facility in remote locations. To demonstrate the radar’s capabilities, we also present examples of its use in three recent field campaigns and its ongoing observations at the Chilbolton Facility for Atmospheric and Radio Research (CFARR).

Analyses of the radar-observed structure and derived rainfall statistics of warm-season convection developing columns of enhanced positive differential reflectivity ZDR over England’s southwest peninsula are presented here. Previous observations of ZDR columns in developing cumulonimbus clouds over England were rare. The observations presented herein suggest otherwise, at least in the southwesterly winds over the peninsula. The results are the most extensive of their kind in the United Kingdom; the data were collected using the National Centre for Atmospheric Science dual-polarization X-band radar (NXPol) during the Convective Precipitation Experiment (COPE). In contrast to recent studies of ZDR columns focused on deep clouds that developed in high-instability environments, the COPE measurements show relatively frequent ZDR columns in shallower clouds, many only 4–5 km deep. The presence of ZDR columns is used to infer that an active warm rain process has contributed to precipitation evolution in convection deep enough for liquid and ice growth to take place. Clouds with ZDR columns were identified objectively in three COPE deployments, with both discrete convection and clouds embedded in larger convective complexes developing columns. Positive ZDR values typically extended to 1–1.25 km above 0°C in the columns, with ZDR ≥ 1 dB sometimes extending nearly 4 km above 0°C. Values above 3 dB typically occurred in the lowest 500 m above 0°C, with coincident airborne measurements confirming the presence of supercooled raindrops. Statistical analyses indicated that the convection that produced ZDR columns was consistently associated with the larger derived rainfall rates when compared with the overall convective population sampled by the NXPol during COPE.

Background:\ud Aromatase inhibitors improve disease-free survival compared with tamoxifen in postmenopausal women with hormone receptor–positive breast cancer. The Tamoxifen and Exemestane Adjuvant Multinational (TEAM) trial compared exemestane monotherapy with sequential therapy of tamoxifen followed by exemestane. The trial failed to show a statistically significant difference between treatment arms. A robust translational program was established to investigate predictive biomarkers.\ud \ud Methods:\ud A tissue microarray was retrospectively constructed using a subset of patient tissues (n = 4631) from the TEAM trial (n = 9766). Immunohistochemistry was performed for biomarkers, classed into three groups: MAPK pathway, NF-kappa B pathway, and estrogen receptor (ER) phosphorylation. Expression was analyzed for association with relapse-free survival (RFS) at 2.5 and 10 years and treatment regimen using Kaplan-Meier curves and log-rank analysis. All statistical tests were two-sided.\ud \ud Results:\ud In univariate analysis, ER167 (hazard ratio [HR] = 0.71, 95% confidence interval [CI] = 0.59 to 0.85, P < .001), IKKα (HR = 0.74, 95% CI = 0.60 to 0.92, P = .005), Raf-1338 (HR = 0.64, 95% CI = 0.52 to 0.80, P < .001), and p44/42 MAPK202/204 (HR = 0.77, 95% CI = 0.64 to 0.92, P = .004) were statistically significantly associated with improved RFS at 10 years in patients receiving sequential therapy. Associations were strengthened when IKKα, Raf-1338, and ER167 were combined into a cumulative prognostic score (HR = 0.64, 95% CI = 0.52 to 0.77, P

It is increasingly appreciated that host factors within the tumor center and microenvironment play a key role in dictating colorectal cancer (CRC) outcomes. As a result, the metastatic process has now been defined as a result of epithelial-mesenchymal transition (EMT). Establishment of the role of EMT within the tumor center and its effect on the tumor microenvironment would be beneficial for prognosis and therapeutic intervention in CRC. The present study assessed five immunohistochemical EMT markers within the tumor center on a 185 Stage II/III CRC patient tissue microarray. In 185 patients with CRC, cytoplasmic snail (HR 1.94 95% confidence interval [CI] 1.15-3.29, p = 0.012) and a novel combined EMT score (HR 3.86 95% CI 2.17-6.86, p0.001) were associated with decreased cancer-specific survival. The combined EMT score was also associated with increased tumor budding (p = 0.046), and systemic inflammation (p = 0.007), as well as decreased memory T-cells within the stroma (p = 0.030) and at the invasive margin (p = 0.035). Furthermore, the combined EMT score was associated with cancer-specific survival independent of TNM-stage (HR 4.12 95% CI 2.30-7.39, p0.001). In conclusion, a novel combined EMT score stratifies patient's survival in Stage II/III CRC and associates with key factors of tumor metastasis. Therefore, the combined EMT score could be used to identify patients at risk of micrometastases and who may benefit from standard adjuvant therapy, potentially in combination with EMT blockade.

N-glycosylation profoundly affects the biological stability and function of therapeutic proteins, which explains the recent interest in glycoengineering technologies as methods to develop biobetter therapeutics. In current manufacturing processes, N-glycosylation is host-specific and remains difficult to control in a production environment that changes with scale and production batches leading to glycosylation heterogeneity and inconsistency. On the other hand, in vitro chemoenzymatic glycan remodeling has been successful in producing homogeneous pre-defined protein glycoforms, but needs to be combined with a cost-effective and scalable production method. An efficient chemoenzymatic glycan remodeling technology using a plant expression system that combines in vivo deglycosylation with an in vitro chemoenzymatic glycosylation is described. Using the monoclonal antibody rituximab as a model therapeutic protein, a uniform Gal2GlcNAc2Man3GlcNAc2 (A2G2) glycoform without α-1,6-fucose, plant-specific α-1,3-fucose or β-1,2-xylose residues was produced. When compared with the innovator product Rituxan®, the plant-made remodeled afucosylated antibody showed similar binding affinity to the CD20 antigen but significantly enhanced cell cytotoxicity in vitro. Using a scalable plant expression system and reducing the in vitro deglycosylation burden creates the potential to eliminate glycan heterogeneity and provide affordable customization of therapeutics' glycosylation for maximal and targeted biological activity. This feature can reduce cost and provide an affordable platform to manufacture biobetter antibodies.

Making high-resolution observations of heavy rainfall from cumulonimbus clouds involves choosing a good location where convection occurs frequently and with some degree of predictability so that aircraft do not have to chase clouds, and radars and other ground-based instruments can be optimally placed. It also requires significant resources, international collaboration and a good deal of serendipity. This paper describes several observational campaigns that were designed to understand relatively small-scale convective clouds that produce heavy precipitation, and presents results that have been found over the years. It complements the companion papers in this special issue by giving an observational perspective. It is by no means comprehensive and is limited to cumulonimbus clouds. The Convective Storm Initiation Project (CSIP) was designed to understand the phenomena responsible for the initiation of convection in the maritime environment of southern England. There was a similar objective in the Convective Orographically-induced Precipitation Study (COPS), but for the complex terrain of the Black Forest mountains, Germany, and the Vosges Mountains, France. The recent Convective Precipitation Experiment (COPE) was designed to understand the entire life cycle of convective clouds that formed along sea-breeze convergence lines in the southwestern peninsula of England. Its particular focus was on the dynamics and microphysics of the convective clouds. Examples of observations from these and other field campaigns designed to study the initiation of convection, cumulonimbus clouds and convective precipitation are presented, with a view to demonstrating how they contribute to a physical understanding of the phenomena and the processes involved.

// Lindsay Bennett 1, 3 , Elizabeth A. Mallon 2 , Paul G. Horgan 3 , Andrew Paul 4 , Donald C. McMillan 3 and Joanne Edwards 1 1 Wolfson Wohl Cancer Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, Scotland, United Kingdom 2 Department of Pathology, Queen Elizabeth University Hospital, Glasgow, Scotland, United Kingdom 3 Academic Unit of Surgery, School of Medicine, University of Glasgow, Glasgow Royal Infirmary, Glasgow, Scotland, United Kingdom 4 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, Scotland, United Kingdom Correspondence to: Joanne Edwards, email: joanne.edwards@glasgow.ac.uk Keywords: breast cancer, NF-κB, tumour microenvironment, survival Received: November 17, 2016 Accepted: February 12, 2017 Published: March 09, 2017 ABSTRACT The aim of the present study was to examine the relationship between tumour NF-κB activation, tumour microenvironment including local inflammatory response (LIR) and cancer-specific survival in patients with operable ductal breast cancer. Immunohistochemistry (tissue microarray of 376 patients) and western blotting (MCF7 and MDA-MB-231 breast cancer cells) was performed to assess expression of key members of the canonical NF-κB pathway (inhibitory kappa B kinase (IKKβ) and phosphorylated p65 Ser-536 (p-p65)). Following silencing of IKKβ, cell viability and apoptosis was assessed in both MCF7 and MDA-MB-231 cell lines. P-p65 was associated with cancer-specific survival (CSS) (nuclear P=0.042 and total P=0.025). High total p-p65 was associated with increase grade tumour grade (P=0.010), ER positivity (P=0.023), molecular subtype (P=0.005), lower Klintrup-Makinen grade (P=0.013) and decreased CD138 count (P=0.032). On multivariate analysis, total p-p65 expression independently associated with poorer CSS (P=0.020). In vitro work demonstrated that the canonical NF-κB pathway was inducible by exposure to TNFα in ER-positive MCF7 cells and to a lesser extent in ER-negative MDA-MB-231 cells. Reduction of IKKβ expression by siRNA transfection increased levels of apoptosis and reduced cell viability in both MCF7 (P= 0.001, P=0.002, respectively). This is consistent with the hypothesis that canonical IKKβ-NF-κB signalling drives tumour survival. These results suggest that activation of the canonical NF-κB pathway is an important determinant of poor outcome in patients with invasive ductal breast cancer.

The original version of this article contained an author name error. In this article, Katrina Madella has been corrected to Katrina Maddela.

The circadian clock regulates the expression of many genes involved in a wide range of biological functions through output pathways such as mitogen-activated protein kinase (MAPK) pathways. We demonstrate here that the clock regulates the phosphorylation, and thus activation, of the MAPKs MAK-1 and MAK-2 in the filamentous fungus Neurospora crassa . In this study, we identified genetic targets of the MAK-1 pathway, which is homologous to the cell wall integrity pathway in Saccharomyces cerevisiae and the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in mammals. When MAK-1 was deleted from Neurospora cells, vegetative growth was reduced and the transcript levels for over 500 genes were affected, with significant enrichment for genes involved in protein synthesis, biogenesis of cellular components, metabolism, energy production, and transcription. Additionally, of the ∼500 genes affected by the disruption of MAK-1, more than 25% were previously identified as putative clock-controlled genes. We show that MAK-1 is necessary for robust rhythms of two morning-specific genes, i.e., ccg-1 and the mitochondrial phosphate carrier protein gene NCU07465. Additionally, we show clock regulation of a predicted chitin synthase gene, NCU04352, whose rhythmic accumulation is also dependent upon MAK-1. Together, these data establish a role for the MAK-1 pathway as an output pathway of the circadian clock and suggest a link between rhythmic MAK-1 activity and circadian control of cellular growth.

The aim of our study was to examine the relationship between tumour IKKα expression and breast cancer recurrence and survival. Immunohistochemistry was employed in a discovery and a validation tissue microarray to assess the association of tumour IKKα expression and clinico-pathological characteristics. After siRNA-mediated silencing of IKKα, cell viability and apoptosis were assessed in MCF7 and MDA-MB-231 breast cancer cells. In both the discovery and validation cohorts, associations observed between IKKα and clinical outcome measures were potentiated in oestrogen receptor (ER) positive Luminal A tumours. In the discovery cohort, cytoplasmic IKKα was associated with disease-free survival (p = 0.029) and recurrence-free survival on tamoxifen (p 0.001) in Luminal A tumours. Nuclear IKKα and a combination of cytoplasmic and nuclear IKKα (total tumour cell IKKα) were associated with cancer-specific survival (p = 0.012 and p = 0.007, respectively) and recurrence-free survival on tamoxifen (p = 0.013 and p 0.001, respectively) in Luminal A tumours. In the validation cohort, cytoplasmic IKKα was associated with cancer-specific survival (p = 0.023), disease-free survival (p = 0.002) and recurrence-free survival on tamoxifen (p = 0.009) in Luminal A tumours. Parallel experiment with breast cancer cells in vitro demonstrated the non-canonical NF-κB pathway was inducible by exposure to lymphotoxin in ER-positive MCF7 cells and not in ER-negative MDA-MB-231 cells. Reduction in IKKα expression by siRNA transfection increased levels of apoptosis and reduced cell viability in MCF7 but not in MDA-MB-231 cells. IKKα is an important determinant of poor outcome in patients with ER-positive invasive ductal breast cancer and thus may represent a potential therapeutic target.

// Antonia K. Roseweir 1, 2 , Arfon G.M.T. Powell 2, 3 , Lindsay Bennett 2 , Hester C. Van Wyk 1 , James Park 1 , Donald C McMillan 1 , Paul G. Horgan 1 , Joanne Edwards 2 1 Academic Unit of Surgery, School of Medicine, University of Glasgow, Royal Infirmary, Glasgow, United Kingdom 2 Unit of Experimental Therapeutics, Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Glasgow, United Kingdom 3 Division of Cancer and Genetics, Cardiff University, Heath Park, Cardiff, United Kingdom Correspondence to: Antonia K. Roseweir, email: antonia.roseweir@glasgow.ac.uk Keywords: colorectal cancer, PTEN, Akt, COX-2, inflammation Received: June 30, 2016 Accepted: September 04, 2016 Published: September 20, 2016 ABSTRACT In patients with colorectal cancer (CRC), local and systemic inflammatory responses have been extensively reported to associate with cancer survival. However, the specific signalling pathways responsible for inflammatory responses are not clear. The PTEN/Akt pathway is a plausible candidate as it may play a role in mediating inflammation via COX-2, and has been associated with cancer progression. This study therefore examined the relationship between tumour PTEN/Akt/COX-2 expression, inflammatory responses and survival in CRC patients using a tissue microarray. In 201 CRC patients, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (12.0yrs v 7.3yrs, P =0.032), poorer differentiation ( P =0.032), venous invasion ( P =0.008) and peritoneal involvement ( P =0.004). Patients were stratified for peri-nuclear expression of COX-2 to examine associations with inflammatory responses. In patients with absent peri-nuclear COX-2 expression, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (11.9yrs v 5.4yrs , P =0.001), poorer differentiation ( P =0.018), venous invasion ( P =0.003) and peritoneal involvement ( P =0.001). However, no associations were seen with either the local or systemic inflammatory responses. In CRC patients, tumour-specific PTEN/Akt pathway activation was significantly associated with poorer CSS, particularly when peri-nuclear COX-2 expression was absent. However, activation of the PTEN/Akt pathway appears not to be responsible for the regulation of inflammatory responses.

Background: Cell line models suggest that activation of NFκB is associated with progression of prostate cancer. This pathway may be a therapeutic target if these observations translate to clinical specimens. Methods: Immunohistochemistry measured NFκBp65 (p65), NFκBp65 nuclear localisation signal (NLS), NFκBp65 phosphorylated at ser 276 (p65ser276), NFκBp65 phosphorylated at ser 536 (p65ser536), IκBα phosphorylated at ser 32/36 (pIκBαser32/36) and MMP-9 protein expression in 61 matched hormone naive prostate cancer (HNPC) and castrate-resistant prostate cancer (CRPC) tumours. Animal and cell models were used to investigate the role of NFκB inhibition in prostate carcinogenesis. Results: In HNPC tumours, NLS expression significantly associated with a shorter time to disease recurrence and disease-specific death. In CRPC tumours p65, pIκBαser32/36 and MMP-9 expression significantly associated with shorter time to death from disease recurrence and shorter disease-specific death. MMP-9 and pIκBαser32/36 expression significantly associated with metastases at recurrence and were independent of Gleason sum and prostate-specific antigen at recurrence. Expression of phosphorylated Akt was associated with increased p65 activation in mouse models and inhibition of NFκB in LNCaP cells significantly reduced cellular proliferation and induced apoptosis. Conclusion: These results provide further evidence that the NFκB pathway could be exploited as a target for CRPC.

Doppler-On-Wheels radar observations made during the Convective and Orographically-induced Precipitation Study (COPS) on 12 August 2007 showed that precipitating clouds only developed between the north–south orientated Murg and Nagold Valleys of the northern Black Forest. The clouds produced moderate precipitation. The Weather Research and Forecasting (WRF) model run at 700 m horizontal resolution in the inner domain was able to simulate the location of the precipitation. Insight is therefore gained into the physical mechanisms responsible for the initiation and development of the convection. Convergence lines resulting from thermally driven flows constrained the initial location of the convection within warm and moist cores produced by heating on elevated surfaces. The heaviest precipitation was later produced by secondary convection, which was initiated within the cores at the boundary between cold-pool outflows and thermal flows. Copyright © 2011 Royal Meteorological Society

Within the framework of the international field campaign COPS (Convective and Orographically induced Precipitation Study), a large suite of state-of-the-art meteorological instrumentation was operated, partially combined for the first time. This includes networks of in situ and remote-sensing systems such as the Global Positioning System as well as a synergy of multi-wavelength passive and active remote-sensing instruments such as advanced radar and lidar systems. The COPS field phase was performed from 01 June to 31 August 2007 in a low-mountain area in southwestern Germany/eastern France covering the Vosges mountains, the Rhine valley and the Black Forest mountains. The collected dataset covers the entire evolution of convective precipitation events in complex terrain from their initiation, to their development and mature phase until their decay. Eighteen Intensive Observations Periods with 37 operation days and eight additional Special Observations Periods were performed, providing a comprehensive dataset covering different forcing conditions. In this article, an overview of the COPS scientific strategy, the field phase, and its first accomplishments is given. Highlights of the campaign are illustrated with several measurement examples. It is demonstrated that COPS research provides new insight into key processes leading to convection initiation and to the modification of precipitation by orography, in the improvement of quantitative precipitation forecasting by the assimilation of new observations, and in the performance of ensembles of convection-permitting models in complex terrain.

The canonical NF-κB pathway regulates the transcription of many genes which may be involved in processes such as inflammation and proliferation, suggesting a potential link between the tumor, the tumor microenvironment and cancer-specific survival in colorectal cancer patients. Immunohistochemistry was used to assess the expression of the upstream kinase TAK1/pTAK1 and four members of the canonical NF-κB pathway in tissue microarrays. Protein expression was determined using the weighted histoscore method. BRaf status, tumor stroma percentage, local inflammation, systemic inflammation and cancer-specific survival were examined in patients with colorectal cancer. Cytoplasmic IKKβ was significantly associated with the inflammatory cell infiltrate (p=0.0015), systemic inflammation (p=0.03) and cancer-specific survival (p=0.046). In contrast, no significant association was found with the other three members. Cytoplasmic pTAK1 was associated with the tumor microenvironment (p=0.045) and cancer-specific survival (p=0.032). On multivariate analysis only cytoplasmic IKKβ associated with survival (HR1.75, 95%CI 1.05-291, p=0.033) and this was independent of TNM stage and markers of the tumor microenvironment. When cytoplasmic IKKβ was stratified with BRaf status (wild type (n=151) or mutant (n=48)), it was no longer associated with cancer-specific survival. However, when cytoplasmic pTAK1 was stratified with BRaf status its association with cancer-specific survival was strengthened. Cytoplasmic pTAK1 was significantly associated with cancer-specific survival in patients with wild type BRaf (p=0.014). In patients with BRaf mutations there was no significant association with cancer-specific survival (p=0.105), however the numbers were low and this is currently being investigated in a larger cohort. The results of the present study show that high expression of cytoplasmic IKKβ was associated with decreased cancer-specific survival and with markers of the tumor microenvironment in patients with colorectal cancer. Expression of cytoplasmic pTAK1 was associated with cancer-specific survival and this was enhanced in patients with tumors expressing wild type BRaf. Citation Format: Jean A. Quinn, Lindsay Bennett, Meera Patel, Lynette Loi, Mikaela Frixou, Antonia Roseweir, James H. Park, Paul G. Horgan, Donald C. McMillan, Joanne Edwards. The relationship between members of the canonical NF-κB pathway, components of the tumor microenvironment and cancer-specific survival in colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4070.

The Boundary Layer Evolution (BLE) missions of the International H2O Project (IHOP_2002) were designed to provide comprehensive observations of the distribution of water vapor in the quiescent boundary layer and its evolution during the early morning. The case study discussed in this paper presents detailed observations of the development of the boundary layer from before sunrise through to the period of growth of the mature convective boundary layer (CBL) during the 14 June 2002 BLE mission. The large number of remote sensing platforms, including the multiple instruments collocated at the Homestead Profiling Site, provided a detailed set of measurements of the growth and structure of the CBL. The observations describe the classic evolution of a daytime CBL, beginning with a shallow nocturnal boundary layer (NBL) below the remnants of the previous day’s mixed layer, or residual layer. The vertical distribution of humidity in these layers during the early morning was affected by advection of dry air and by gravity waves. About an hour after sunrise a CBL developed, and gradually deepened with time as it mixed out the NBL and residual layer. The growth of the top of the CBL was particularly well observed because of the strong vertical gradients in temperature, humidity, and aerosol concentration. As the CBL deepened and the average CBL wind speed decreased, the mode of convective organization evolved from horizontal convective rolls to open-celled convection. A unique set of detailed measurements of the structure of the open cells was obtained from multiple instruments including the Doppler-on-Wheels radar, the Mobile Integrated Profiling System wind profiler, and the Scanning Raman lidar. They showed the relationship between open cells, thermals, mantle echoes, and the CBL top.

BPV-1 and less commonly BPV-2 are associated with the pathogenesis of equine skin tumours termed sarcoids. We recently documented the transcriptional changes that are induced by BPV-1 in equine fibroblasts using microarray analyses. TLR4 expression was found to be significantly down-regulated by BPV-1. In the present study, we show that TLR4 expression is significantly decreased following the exogenous expression of BPV-1 E2 and E7 in primary equine fibroblasts. The results were confirmed by the demonstration of increased TLR4 expression following siRNA suppression of BPV-1 E2 and E7 viral gene expression. These data imply that BPV-1 is able to subvert the innate immune response by downregulation of TLR4.

Students’ perceptions of school safety and experiences with bullying were examined in a large Canadian cohort of 5,493 girls and 5,659 boys in Grades 4 to 12. Results indicate notable differences in when and where students felt safe based on their own perceptions of safety and their own experiences with bullying, particularly across elementary and secondary schools. For elementary students, especially those involved in bullying, the playground/school yard and outside recess/break time were particularly hazardous, whereas for secondary students involved in bullying, the hallways, school lunchroom/cafeteria, and outside recess/break were considered especially dangerous. The commonality across student-identified unsafe areas is that they tend to not be well supervised by school personnel. Accordingly, the present results underscore the need to increase adult supervision in areas in which an overwhelming majority of students report feeling unsafe.

Circadian clocks and mitogen-activated protein kinase (MAPK) signaling pathways are fundamental features of eukaryotic cells. Both pathways provide mechanisms for cells to respond to environmental stimuli, and links between them are known. We recently reported that the circadian clock in Neurospora crassa regulates daily rhythms in accumulation of phosphorylated, and thus active, OS-2 MAPK, a relative of mammalian p38 MAPK, when cells are grown in constant conditions. In the absence of acute stress, rhythmically activated MAPK then signals to downstream effector molecules to regulate rhythmic expression of target genes of the pathway. Clock regulation of MAPK signaling pathways provides a mechanism to coordinately control major groups of genes such that they peak at the appropriate times of day to provide a growth and survival advantage to the organism by anticipating stresses. MAPK pathways are well known for their role in cell proliferation and tumor suppression. New evidence reveals that some mammalian clock components also function as tumor suppressors and rhythms in phospho-MAPK have been observed in higher eukaryotes. Thus, the role of the clock in regulation of the activity of MAPK pathways provides important clues into the function of the circadian clock as a tumor suppressor.

Rapid, large-scale manufacture of medical countermeasures can be uniquely met by the plant-made-pharmaceutical platform technology. As a participant in the Defense Advanced Research Projects Agency (DARPA) Blue Angel project, the Caliber Biotherapeutics facility was designed, constructed, commissioned and released a therapeutic target (H1N1 influenza subunit vaccine) in

In Neurospora crassa, FRQ, WC-1, and WC-2 proteins comprise the core circadian FRQ-based oscillator that is directly responsive to light and drives daily rhythms in spore development and gene expression. However, physiological and biochemical studies have demonstrated the existence of additional oscillators in the cell that function in the absence of FRQ (collectively termed FRQ-less oscillators [FLOs]). Whether or not these represent temperature-compensated, entrainable circadian oscillators is not known. The authors previously identified an evening-peaking gene, W06H2 (now called clock-controlled gene 16 [ ccg-16]), which is expressed with a robust daily rhythm in cells that lack FRQ protein, suggesting that ccg-16 is regulated by a FLO. In this study, the authors provide evidence that the FLO driving ccg-16 rhythmicity is a circadian oscillator. They find that ccg-16 rhythms are generated by a temperature-responsive, temperature-compensated circadian FLO that, similar to the FRQ-based oscillator, requires functional WC-1 and WC-2 proteins for activity. They also find that FRQ is not essential for rhythmic WC-1 protein levels, raising the possibility that this WCFLO is involved in the generation of WC-1 rhythms. The results are consistent with the presence of 2 circadian oscillators within Neurospora cells, which the authors speculate may interact with each other through the shared WC proteins.

Recent severe weather events have prompted the European scientific community to assess the current understanding of convective processes with a view to more detailed and accurate forecasting. The initial development of convective cells remains one of the least understood aspects and one in which limited research has taken place. The important processes can be split into three main areas: boundary-layer forcing, upper-level forcing and secondary generation. This paper is a review of the mechanisms responsible for the initiation of precipitating convection in the United Kingdom; i.e. why convective clouds form and develop into precipitating clouds in a particular location. The topography of the United Kingdom has a large influence on the initiation of convection. Boundary-layer forcings determine the specific location where convection is triggered within larger regions of potential instability. These latter regions are created by mesoscale or synoptic-scale features at a higher level such as dry intrusions and mesoscale vortices. Second-generation cells are those formed by the interaction of outflow from convective clouds with the surrounding environmental air. Large, long-lived thunderstorm complexes can develop when new cells are repeatedly triggered on one side of the system. Current and future field campaigns along with the development of high-resolution modelling will enable these processes to be investigated in more detail than has previously been achieved. © Royal Meteorological Society, 2006. Contributions by P. A. Clark and M. E. B. Gray are Crown Copyright.

631 Background: In patients with colorectal cancer (CRC) the local and systemic inflammatory responses (LIR and SIR) are important determinants of disease progression. The present study examines the association of transcription factor RelB, a key member of the non-canonical NF-κB pathway and its association with LIR and SIR in patients undergoing resection of CRC. Methods: Patients with stage I-III CRC who underwent curative resection in a single institution and were in a previously constructed tissue microarray were studied. IHC was utilised to examine cytoplasmic and nuclear RelB expression. The relationship between RelB, clinicopathological characteristics, LIR (Klintrup-Mäkinen (KM) grade, CD3+ and CD8+T-cell density), SIR and cancer-specific survival (CSS) was examined. Results: 208 patients were included in the analysis. Cytoplasmic RelB (cyto-RelB) was associated with nuclear RelB ( p=0.006). High expression of cyto-RelB was associated with MMR competence ( p=0.010) but not with TNM stage ( p=0.468), venous invasion ( p=0.973), tumour budding ( p=0.068), tumour necrosis ( p=0.786), tumour cell proliferation ( p=0.907), BRAF V600E mutation ( p=0.585) or administration of adjuvant chemotherapy ( p=0.853). High cyto-RelB was inversely associated with mGPS (mGPS >1: low cyto-RelB – 19% vs. high cyto-RelB – 8%, p=0.017). Also, cyto-RelB was inversely associated with tumour inflammatory cell infiltrate at the margin, Klintrup-Mäkinen grade ( p=0.059), (CD3+ p=0.010, CD8+ p=0.007) and in the tumour (CD3+ p=0.002) and a trend with tumour stroma percentage ( p=0.079). High expression of cyto-RelB was not significantly associated with CSS ( p=0.052). Conclusions: In patients undergoing CRC resection, high expression of cyto-RelB was associated with an adverse host local inflammatory response. Up-regulation of the non-canonical NF- κB pathway may be an important mechanism whereby the tumour deregulates local inflammatory responses and evades host immunosurveillance. Further investigation of inflammation based signal transduction pathways in patients with colorectal cancer is warranted.

A case study of orographic convection initiation (CI) that occurred along the eastern slopes of the Vosges Mountains in France on 6 August 2007 during the Convective and Orographically-Induced Precipitation Study (COPS) is presented. Global positioning system (GPS) receivers and two Doppler on Wheels (DOW) mobile radars sampled the preconvective and storm environments and were respectively used to retrieve three-dimensional tomographic water vapor and wind fields. These retrieved data were supplemented with temperature, moisture, and winds from radiosondes from a site in the eastern Rhine Valley. High-resolution numerical simulations with the Weather Research and Forecasting (WRF) Model were used to further investigate the physical processes leading to convective precipitation. This unique, time-varying combination of derived water vapor and winds from observations illustrated an increase in low-level moisture and convergence between upslope easterlies and downslope westerlies along the eastern slope of the Vosges Mountains. Uplift associated with these shallow, colliding boundary layer flows eventually led to the initiation of moist convection. WRF reproduced many features of the observed complicated flow, such as cyclonic (anticyclonic) flow around the southern (northern) end of the Vosges Mountains and the east-side convergent flow below the ridgeline. The WRF simulations also illustrated spatial and temporal variability in buoyancy and the removal of the lids prior to convective development. The timing and location of CI from the WRF simulations was surprisingly close to that observed.

Raf Kinase inhibitory protein (RKIP) is a well-established metastasis suppressor that is frequently downregulated in aggressive cancers. The impact of RKIP and its phosphorylated form on disease-free survival (DFS) and other clinicopathological parameters in breast cancer is yet to be discovered. To this end, we examined RKIP expression in 3 independent breast cancer cohorts. At the Protein level, loss or reduced total RKIP expression was associated with large-sized tumors characterized by high proliferative index, high-grade and diminished estrogen (ER) and progesterone receptor expression. Loss or diminution of RKIP expression was significantly associated with shorter DFS in all cohorts. Moreover, the complete loss of p-RKIP was an independent prognostic factor using multivariate analysis in operable invasive ductal breast cancer. We show for the first time that ER, partly, drives RKIP expression through MTA3-Snail axis. Consistent with this finding, we found that, at the mRNA level, RKIP expression varied significantly across the different molecular subtypes of breast cancer with the Luminal (ER+) subtype expressing high levels of RKIP and the more aggressive Claudin-low (ER-) subtype, which depicted the highest epithelial to mesenchymal transition (EMT) registered the lowest RKIP expression levels. In conclusion, loss of expression/diminution of RKIP or its phosphorylated form is associated with poor diseases-free survival in breast cancer. Determining the expression of RKIP and p-RKIP adds significant prognostic value to the management and subtyping of this disease.

We previously reported that AR phosphorylation at serine 213 was associated with poor outcome and may contribute to prostate cancer development and progression. This study investigates if specific AR phosphorylation sites have differing roles in the progression of hormone naïve prostate cancer (HNPC) to castrate resistant disease (CRPC). A panel of phosphospecific antibodies were employed to study AR phosphorylation in 84 matched HNPC and CRPC tumours. Immunohistochemistry measured Androgen receptor expression phosphorylated at serine residues 94 (pAR94), 308 (pAR308), 650(pAR650) and 791(pAR791). No correlations with clinical parameters were observed for pAR94 or pAR650 in HNPC or CRPC tumours. In contrast to our previous observation with serine 213, high pAR308 is significantly associated with a longer time to disease specific death (p= 0.011) and high pAR791 expression significantly associated with a longer time to disease recurrence (p= 0.018) in HNPC tumours and longer time to death from disease recurrence (p= 0.040) in CRPC tumours. This observation in CRPC tumours was attenuated in high apoptotic tumours (p= 0.022) and low proliferating tumours (p= 0.004). These results demonstrate that understanding the differing roles of AR phosphorylation is necessary before this can be exploited as a target for castrate resistant prostate cancer.

The canonical NF-κB pathway regulates transcription of a wide range of genes involved in various processes including inflammation and proliferation, suggesting that it might provide a link between the systemic inflammatory response (SIR), local inflammatory response (LIR) and tumor signalling, the present study was to examine the relationship between SIR, tumor NF-κB pathway, tumor microenvironment including LIR and cancer specific survival in patients with colorectal cancer Immunohistochemistry of inhibitory kappa β kinase (IKK)-β, p65 (Rel A) and phosphorylated p65 (p-p65) was performed using colorectal cancer tissue microarray of 151 patients and the weight histoscore method was employed to assess protein expression. The relationship between these proteins, BRAF status, tumor stroma percentage, local inflammation, systemic inflammation, and cancer-specific survival were examined. In addition, western blotting was performed to assess expression of key members of the canonical NF-κB pathway following stimulation with either TNFα or IL-1 in BRAF wildtype (T84) and BRAF V600E mutated cells (HT-29). Expression of neither p65 nor p-p65, at any cellular location, was associated with clinical outcome measures. However a significant association between cancer specific survival and cytoplasmic IKKβ (p = 0.015), was observed and 10 year survival stratified from 78% to 59%. In addition, when patients were stratified by BRAF status the association with cancer specific survival and cytoplasmic IKKβ was upheld in those patients with wild type BRAF (p = 0.014) but negated in those with mutated BRAF (p = 0.287). Cytoplasmic IKKβ was significantly associated with venous invasion (p = 0.024), immunoscore (p = 0.023), Klintrup-Makinen grade (p = 0.050) and Glasgow microenvironment score. Total cell IKKβ expression was also associated with cancer specific survival, (p = 0.015) and 10 year survival was stratified from 73% to 53%. Total cell IKKβ was significantly associated with TNM (p = 0.004), tumor differentiation (p = 0.001), venous invasion (p = 0.011), necrosis (p = 0.003) and tumor stoma percentage (p = 0.048). In vitro, the canonical NF-κB pathway was induced in colon cancer cell lines T84 and HT-29 by exposure to either TNFα or IL-1 (p-p65 levels increased at 15 min), IKKβ expression remained constant. In colorectal cancer patients, cytoplasmic and total cell IKKβ expression is associated with cancer specific survival and this was related to tumor microenviroment and LIR but not the SIR. Interactions between stromal and NF-κB pathway may be an important method by which disease progression occurs. Citation Format: Jean A. Quinn, Lindsay Bennett, Antonia Roseweir, James H. Park, Paul G. Horgan, Donald C. McMillan, Joanne Edwards. The relationship between members of the canonical NF-κB pathway, components of the microenvironment and survival in patients with colorectal cancer [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B125.

MAPK signal transduction pathways are important regulators of stress responses, cellular growth, and differentiation. In Neurospora, the circadian clock controls rhythms in phosphorylation of the p38-like MAPK (OS-2); however, the mechanism for this regulation is not known. We show that the WCC, a transcription factor and clock component, binds to the os-4 MAPKKK promoter in response to light and rhythmically in constant darkness, peaking in the subjective morning. Deletion of the WCC binding sites in the os-4 promoter disrupts both os-4 mRNA and OS-2 phosphorylation rhythms. The clock also indirectly regulates rhythmic expression of the histidyl-phosphotransferase gene, hpt-1, which peaks in the evening. Anti-phase expression of positive (OS-4) and negative (HPT-1) MAPK pathway regulators likely coordinate to enhance rhythmic MAPK activation to prepare cells to respond to osmotic stress during the day in the natural environment. Consistent with this idea, we show that wild type cells have a clock-dependent morning kinetic advantage in glycerol accumulation after salt stress as compared to evening treatment. Thus, circadian transcriptional control of MAPK pathway components leads to striking time-of-day-specific effects on the signaling status and physiological response of the pathway.

Copyright @ 2007 AMS The Convective Storm Initiation Project (CSIP) is an international project to understand precisely where, when, and how convective clouds form and develop into showers in the mainly maritime environment of southern England. A major aim of CSIP is to compare the results of the very high resolution Met Office weather forecasting model with detailed observations of the early stages of convective clouds and to use the newly gained understanding to improve the predictions of the model. A large array of ground-based instruments plus two instrumented aircraft, from the U.K. National Centre for Atmospheric Science (NCAS) and the German Institute for Meteorology and Climate Research (IMK), Karlsruhe, were deployed in southern England, over an area centered on the meteorological radars at Chilbolton, during the summers of 2004 and 2005. In addition to a variety of ground-based remote-sensing instruments, numerous rawin-sondes were released at one- to two-hourly intervals from six closely spaced sites. The Met Office weather radar network and Meteosat satellite imagery were used to provide context for the observations made by the instruments deployed during CSIP. This article presents an overview of the CSIP field campaign and examples from CSIP of the types of convective initiation phenomena that are typical in the United Kingdom. It shows the way in which certain kinds of observational data are able to reveal these phenomena and gives an explanation of how the analyses of data from the field campaign will be used in the development of an improved very high resolution NWP model for operational use. This work is funded by the National Environment Research Council following an initial award from the HEFCE Joint Infrastructure Fund.

588 Background: Although breast cancer morality rates have fallen, resistance to treatment remains a major problem. The NF-κB pathway regulates transcription of a wide range of genes involved in inflammation, proliferation and apoptosis and are hypothesized to play a role in tumor progression. Methods: Immunohistochemistry was employed to assess expression of members of the NF-κB pathway in a cohort of 544 breast cancer specimens with full clinical follow-up. Antibodies for the p65 subunit, phosphorylated p65 at the serine 536 residue (p-p65 S536), nuclear localization signal of p65 (p65 NLS), IKKα and IKKb had specificity confirmed by both a single band of appropriate size on a western blot and appropriate expression patterns in cell pellets treated with TNFα. Results: When the full cohort was considered patients with tumors that expressed high levels of cytoplasmic IKKα had significantly shorter disease free survival compared to those with low expression (P=0.015). Increased expression of nuclear p-p65 S536 was also associated with shorter disease free survival (P=0.005). By western blot analysis we observed that expression levels of p-p65 S536 were higher in MD-MBA-231 cells which are ER negative compared to MCF-7 cells that are ER positive, suggesting that NF-κB may have a prominent role in ER negative disease. However when the cohort was subdivided, by ER status the association observed for IKKα expression and shorter disease free survival was attenuated in ER positive tumors (p=0.008). In addition this attenuation with the ER positive cohort was also observed for nuclear p-p65 S536 (P=0.002). Other members of the pathway investigated (p65, p65 NLS and IKKb) were not associated with disease free survival. Conclusions: These results suggest that in the clinical setting the NF-kB canonical pathway is associated with poor prognosis in ER positive tumors and not with ER negative tumors as the cell line work would suggest. These results require validation in an independent cohort, but highlight the need to consider subtypes of breast cancer as individual diseases with increasingly different therapeutic targets.

Although therapies exist for breast cancer and mortality rates have fallen, resistance to current therapies remains a major problem. The NF-κB pathways regulate the transcription of a wide range of genes involved in the immune response, inflammation, proliferation and apoptosis. Many of these processes are hallmarks of cancer and NF-κB has been hypothesised to be a link between inflammation and tumourigenesis. The aim of the current study was to investigate the role of both NF-κB pathways in the pathogenesis and recurrence of breast cancer. Immunohistochemistry was employed to assess key components of the canonical (p65 and IKKβ) and non-canonical (NIK, RelB, IKKα) pathways on a TMA of 544 patients with full clinical follow up and clinical information including ER status, subtype (luminal A/B, triple negative, HER2 enriched), necrosis, apoptosis and angiogenesis. Nuclear expression of p-p65 S536 was observed in 81% (336/414) of patients and was associated with angiogenesis (p=0.028) and disease free survival (p=0.003). Those patients with tumours that expressed high levels of p-p65 S536 in the nuclear compartment had significantly shorter disease free survival compared to those with low expression (6.95 years vrs 7.91 years). The cohort was then subdivided by ER status and this observation was attenuated in ER positive tumours (p=0.002, 7.10 years vrs 8.34 years). When split further into subtype this was negated in luminal A tumours (p=0.120) but significance increased in luminal B tumours (p=2.54x10-4, 5.52 years vrs 8.16 years). A differing role was observed for the non-canonical pathway. Cytoplasmic expression of IKKα was observed in 97% (367/378) of patients and was associated with apoptosis (p=0.024) and necrosis (p=0.020) and a shorter disease free survival in those with high expression (p=0.014, 7.16 years vrs 7.64 years). This observation was attenuated in ER positive tumours when the cohort was subdivided by ER status (p=0.008, 7.56 years vrs 8.00 years). When split into subtype, the relationship between cytoplasmic IKKα and disease free survival was negated in all subtypes except luminal A (p=0.015, 7.92 years vrs 8.32 years). Other members of the NF-κB pathways (p65 NLS, IKKβ, NIK and RelB) were not associated with disease free survival. Our results suggest that the NF-κB pathways are involved in promoting recurrence in patients with ER positive tumours and may be involved with development of endocrine resistance. However, each pathway seems to be involved in different subtypes with the canonical pathway associating with recurrence in the luminal B subtype and the non-canonical with the luminal A subtype. Further studies in an independent cohort combined with mechanistic studies are required. These results highlight that patients with ER positive tumours may have different signalling pathways driving recurrence and therefore benefit from different therapeutic strategies. Citation Format: Lindsay Bennett, Clare Orange, Elizabeth Mallon, Julie Doughty, Paul G. Horgan, Andrew Paul, Joanne Edwards. NF-κB pathways are associated with increased recurrence in different subtypes of ER positive breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3468. doi:10.1158/1538-7445.AM2013-3468