Calcium signalling links MYC to NUAK1

NUAK1 is a member of the AMPK-related family of kinases. Recent evidence suggests that NUAK1 is an important regulator of cell adhesion and migration, cellular and organismal metabolism, and regulation of TAU stability. As such, NUAK1 may play key roles in multiple diseases ranging from neurodegeneration to diabetes and metastatic cancer. Previous work revealed a crucial role for NUAK1 in supporting viability of tumour cells specifically when MYC is overexpressed. This role is surprising, given that NUAK1 is activated by the tumour suppressor LKB1. Here we show that, in tumour cells lacking LKB1, NUAK1 activity is maintained by an alternative pathway involving calcium-dependent activation of PKCα. Calcium/PKCα-dependent activation of NUAK1 supports engagement of the AMPK-TORC1 metabolic checkpoint, thereby protecting tumour cells from MYC-driven cell death, and indeed, MYC selects for this pathway in part via transcriptional regulation of PKCα and ITPR. Our data point to a novel role for calcium in supporting tumour cell viability and clarify the synthetic lethal interaction between NUAK1 and MYC

Differential effects of depleting agents on cytoplasmic and nuclear non-protein sulphydryls: a fluorescence image cytometry study.

The intracellular distribution of glutathione (GSH) was measured by a quantitative image cytometry method, using the sulphydryl-reactive agent mercury orange. This readily forms fluorescent adducts with GSH and other non-protein sulphydryls (NPSH), but reacts much more slowly with protein sulphydryls. Under optimum staining conditions mean integrated mercury orange fluorescence per cell was closely correlated with a standard biochemical assay for GSH. Use of the DNA dye DAPI as a counterstain allowed measurement of nuclear NPSH. The mean nuclear-cytoplasmic ratio was 0.57 +/- 0.05. Isolation of nuclei under aqueous conditions resulted in the loss of approximately 90% of mercury orange fluorescence, compared with nuclear fluorescence from intact cells, suggesting that background labelling of protein sulphydryls or other macromolecules is low. Depletion of GSH with N-ethylmaleimide or diethylmaleate decreased mercury orange fluorescence in the nucleus and cytoplasm to a similar extent. In contrast, mercury orange fluorescence in the nucleus was much more resistant to DL-buthionine-S,R-sulphoximine (BSO) depletion than that in the cytoplasm. This finding is compatible with a distinct pool of GSH in the nucleus that is comparatively resistant to BSO depletion. Alternatively, the retention of fluorescence in the nucleus following GSH depletion by BSO treatment might be due to accumulation of cysteine. These findings have implications for cancer treatment since the level of NPSH in the nucleus might be a more important determinant of resistance to DNA-damaging agents than that in cytoplasm. The image cytometry method described here is quantitative, allows a measure of tumour cell heterogeneity and can be applied to small biopsy samples obtained by fine-needle aspiration. Thus it appears suitable for prospective clinical studies in cancer patients, and for monitoring the effects of GSH-depleting agents used as adjuncts to cancer chemotherapy or radiotherapy

Comparison of 99mTc-sestamibi and doxorubicin to monitor inhibition of P-glycoprotein function

P-glycoprotein (Pgp) overexpression is a well-recognized factor in resistance to chemotherapy. Doxorubicin flow cytometry is used to monitor Pgp function in haematological specimens and biopsies from other cancers, and radionuclide imaging with sestamibi has recently shown promise for non-invasive monitoring. In the present study the two methods were directly compared in single-cell suspensions of three variants of the human breast carcinoma cell line MCF7: sensitive MCF7/WT, doxorubicin-selected MCF7/AdrR, and MDR1 -gene-transfected MCF7/BC19 cells with doxorubicin resistance factors of 1, 192, and 14, respectively. Accumulation of sestamibi and mean fluorescence of doxorubicin (5.5 μM) were assessed over 60 min in the presence and absence of Pgp modulators GG918 (0.01 to 0.2 μM) and PSC833 (0.05 to 2.0 μM). Accumulation curves for sestamibi and doxorubicin differed among the cell variants under control conditions, with sestamibi showing a significantly greater difference between WT and resistant cells than doxorubicin. Both GG918 and PSC833 reversed uptake deficits to WT levels for sestamibi in MCF7/BC19 cells and doxorubicin in MCF7/BC19 and MCF7/AdrR cells, but failed to show the same effect for sestamibi in MCF7/AdrR cells (∼30% of MCF7/WT level). Thus, both methods clearly distinguished sensitive from resistant MCF7 variants, with the radionuclide method showing greater sensitivity. © 2001 Cancer Research Campaign http://www.bjcancer.co

Prevalence of Presenting Conditions in Grey Seal Pups (Halichoerus grypus) Admitted for Rehabilitation

A retrospective survey was performed on the presenting conditions of 205 live grey seal pups (Halichoerus grypus) admitted to the Cornish Seal Sanctuary in Gweek, United Kingdom between May 2005 and March 2011. The purpose of the survey was to examine the prevalence of various presenting signs at the sanctuary. The presenting signs were classified into nine non-mutually exclusive categories: ocular disorders, nasal disorders, oral disorders, respiratory disorders, orthopaedic disorders, puncture wounds, abrasions, netting injuries, and onychia. The sex ratio of seal pups in this study was 1.35 males per female. Of the 205 examined for rehabilitation, 22 (10.73%) did not survive to release. 68.78% of grey seal pups presented with puncture wounds, 47.80% with respiratory disorders, 46.34% with ocular disorders, 42.63% malnourished, 36.59% with abrasions, 25.37% with oral disorders, 23.90% with nasal disorders, 11.71% with orthopaedic disorders, 9.27% with onychia, and 3.41% presented with netting injuries. 52% were normothermic, 42% were hyperthermic, and 5% were hypothermic. Associations between gender, outcome of rehabilitation, hospitalisation time and presenting disorders were examined. In addition, admissions rates were found to display seasonality. The results of this study will aid in future preparation of grey seal rehabilitation facilities

Satellite-derived bathymetry in optically complex waters using a model inversion approach and Sentinel-2 data

This study presents an assessment of a model inversion approach to derive shallow water bathymetry in optically complex waters, with the aim of both understanding localised capability and contributing to the global evaluation of Sentinel-2 for coastal monitoring. A dataset of 12 Sentinel-MSI images, in three different study areas along the Irish coast, has been analysed. Before the application of the bathymetric model two atmospheric correction procedures were tested: Deep Water Correction (DWC) and Case 2 Regional Coastal Color (C2RCC) processor. DWC outperformed C2RCC in the majority of the satellite images showing more consistent results. Using DWC for atmospheric correction before the application of the bathymetric model, the lowest average RMSE was found in Dublin Bay (RMSE ¼ 1.60, bias ¼ \u100000 0.51), followed by Mulroy Bay (RMSE ¼ 1.66, bias ¼ 1.30) while Brandon Bay showed the highest average error (RMSE ¼ 2.43, bias ¼ 1.86). However, when the optimal imagery selection was considered, depth estimations with a bias less than 0.1 m and a spread of 1.40 m were achieved up to 10 m. These results were comparable to those achieved by empirical tuning methods, despite not relying on any in situ depth data. This conclusion is of particular relevance as model inversion approaches might allow future modifications in crucial parts of the processing chain leading to improved results. Atmospheric correction, the selection of optimal images (e.g. low turbidity), the definition of suitably limited ranges for the per-pixel occurrence of optical constituents (phytoplankton, CDOM, backscatter) and seabed reflectances, in combination with the understanding of the specifics characteristics at each particular site, were critical steps in the derivation of satellite bathymetry

A clinical prediction rule for diagnosing severe acute respiratory syndrome in the emergency department

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Nanoscale mobility mapping in semiconducting polymer films

This work was supported by grant No 19-12-00066 of the Russian Science Foundation.Local electrical properties of thin films of the polymer PTB7 are studied by conductive atomic force microscopy (C-AFM). Non-uniform nanoscale current distribution in the neat PTB7 film is revealed and connected with the existence of ordered PTB7 crystallites. The shape of local I-V curves is explained by the presence of space charge limited current. We modify an existing semi-empirical model for estimation of the nanoscale hole mobility from our experimental C-AFM measurements. The procedure of nanoscale charge mobility estimation was described and applied to the PTB7 films. The calculated average C-AFM hole mobility is in good agreement with macroscopic values reported for this material. Mapping of nanoscale hole mobility was achieved using the described procedure. Local mobility values, influenced by nanoscale structure, vary more than two times in value and have a root-mean-square value 0.22 × 10−8 m2/(Vs), which is almost 20% from average hole mobility.PostprintPeer reviewe