Small Molecular Inhibitors Block TRPM4 Currents in Prostate Cancer Cells,with Limited Impact on Cancer Hallmark Functions

Transient receptor potential melastatin 4 (TRPM4) is a broadly expressed Ca²⁺ activated monovalent cation channel that contributes to the pathophysiology of several diseases.For this study, we generated stable CRISPR/Cas9 TRPM4 knockout (K.O.) cells from the human prostate cancer cell line DU145 and analyzed the cells for changes in cancer hallmark functions. Both TRPM4-K.O. clones demonstrated lower proliferation and viability compared to the parental cells. Migration was also impaired in the TRPM4-K.O. cells. Additionally, analysis of 210 prostate cancer patient tissues demonstrates a positive association between TRPM4 protein expression and local/metastatic progression. Moreover, a decreased adhesion rate was detected in the two K.O. clones compared to DU145 cells.Next, we tested three novel TRPM4 inhibitors with whole-cell patch clamp technique for their potential to block TRPM4 currents. CBA, NBA and LBA partially inhibited TRPM4 currents in DU145 cells. However, none of these inhibitors demonstrated any TRPM4-specific effect in the cellular assays.To evaluate if the observed effect of TRPM4 K.O. on migration, viability, and cell cycle is linked to TRPM4 ion conductivity, we transfected TRPM4-K.O. cells with either TRPM4 wild-type or a dominant-negative mutant, non-permeable to Na⁺. Our data showed a partial rescue of the viability of cells expressing functional TRPM4, while the pore mutant was not able to rescue this phenotype. For cell cycle distribution, TRPM4 ion conductivity was not essential since TRPM4 wild-type and the pore mutant rescued the phenotype.In conclusion, TRPM4 contributes to viability, migration, cell cycle shift, and adhesion; however, blocking TRPM4 ion conductivity is insufficient to prevent its role in cancer hallmark functions in prostate cancer cells.     

Local synthesis of the phosphatidylinositol-3,4-bisphosphate lipid drives focal adhesion turnover

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Hydro-sedimentology of the Johnstone River estuary

This paper examines the physical consequences of increased catchment sediment yields on the sediment budget and the hydrodynamic setting of the South Johnstone River estuary in North Queensland. A combined study involving hydrological monitoring, assessment of sediment sources, estimation of riverine sediment budget and hydro-sedimentological numerical modelling for estuarine sediment transport is currently underway. Initial field and laboratory observations indicate that the sediment delivery from highly erosion-prone sugar cane cultivations in the tropical catchment has increased dramatically during the last 10 years. This has subsequently given rise to elevated flood levels in both the lower and upper catchment areas, as well as significant modifications to the river bed morphology.     

The particle size characteristics of fluvial suspended sediment: an overview

The particle size characteristics of suspended sediment are of fundamental importance in understanding its role in a variety of environmental processes. Existing knowledge concerning the spatial and temporal variability of the grain size composition of suspended sediment is, however, relatively limited. At the global scale, major contrasts may exist between individual rivers in the calibre of their suspended load and this may be related to a number of controls including climate, catchment geology and basin scale. Any attempt to understand the precise relationship between the grain size characteristics of suspended sediment and those of its source material must also take account of the selectivity of erosion and delivery processes. A local case study undertaken by the authors in the 1500 km² basin of the River Exe in Devon, UK is used to illustrate the considerable spatial variability that may occur within a relatively small area and the complexity of the associated controls.Available evidence concerning the temporal variability of the grain size characteristics of suspended sediment emphasises the diverse patterns of behaviour that may exist and the complexity of the controls involved. In some rivers the sediment may become coarser as flow increases, in others it may become finer, whilst in others it may exhibit a relatively constant grain size composition. Data from the local case study in the Exe basin are again used to highlight the considerable diversity in response to changing discharge that may occur within a relatively small area.Any attempt to understand the dynamics of sediment movement through a river system must also take account of the potential contrast between the ultimate and effective particle size distribution of suspended sediment in response to aggregation. Results from the Exe basin study indicate that even in rivers with relatively low solute concentrations, almost an order of magnitude difference may exist between the median particle size associated with the ultimate and effective grain size distributions.     

Pollen as a chronometer and sediment tracer,Burrinjuck Reservoir,Australia

Pollen analysis is widely used to reconstruct vegetation and land use histories, but can also provide sedimentological information. At Burrinjuck Reservoir, in south-eastern Australia, annual grass pollen peaks are used to distinguish each year's sediment, even when there are no visible laminations. In conjunction with other dating methods, this allows the determination of year by year influxes of all sediment components. Pollen grains in the Burrinjuck sediments are shown to be predominantly waterborne so that they can be used to trace sediment to its source in particular vegetation stands. Pollen concentration and the proportion of damaged pollen might also distinguish sediment eroded from topsoils and that from subsoils. Pollen analysis can thus be used to locate specific erosion events in both time and space.     

Experimental revegetation of the regulated lake Ontojärvi in northern Finland

Water level regulation causes large-scale ecological changes in the littoral areas of lakes in northern Finland. If the summertime water level is raised, intensive erosion processes begin, causing a sudden decline in shore vegetation. The need for shore protection is obvious in areas of high recreational value. At lake Ontojärvi, planting experiments with littoral helophytes and bushes were carried out during the years 1990–92. All the experiments were carried out in the eroded sandy areas, which were partly protected by mechanical barriers. Several plant species were planted on the shore which had been treated with different peat mixtures, etc. The frequencies of the different species were followed monthly. After the first summer, the average survival rates were about 45 % due to the drying of seedlings. A rapid decrease in the survival rates took place during the high water level period in 1991 at which point only 20% of the planted individuals were alive. The best results were obtained for the helophyte Carex rostrata Stokes, of which 30% had survived erosion. Tall willows (Salix phylicifolia L.) were also erosion-resistant with a survival rate of 80%.     

Effects of erosion on ecto- and VA-mycorrhizal inoculum potential of soil following forest fire in southwest Oregon

The Longwood Complex wildfire in the Siskiyou Mountains of southern Oregon in August 1987 created an opportunity to study erosion and its effects on mycorrhizal fungus inoculum potential of a forest soil on steep slopes. As measured by the erosion-bridge method, most erosion occurred in a single, intense storm in December after the fire and amounted to an estimated 2 to 4 cm of surface soil. Captured eroded soil had a higher pH and P and Mg levels than residual soil. Seedlings ofLibocedrus decurrens andPseudotsuga menziesii were planted on eroded plots with additions of captured eroded soil (ET) or pasteurized eroded soil (PET) transferred to the planting holes. After one growing season,Libocedrus seedlings formed nearly 4 times the vesicular-arbuscular mycorrhizae in ET treatments and more than twice as much in PET treatments than in controls. Survival and basal area growth were significantly better in ET than in the other treatments, and both ET and PET produced more seedling shoot growth than did controls.Pseudotsuga seedlings did not differ in measured characteristics between treatments; ectomycorrhiza formation was slight, evidently the result of reduced inoculum potential resulting from the fire.     

PYK2 is overexpressed in chronic lymphocytic leukaemia: A potential new therapeutic target

Chronic Lymphocytic Leukaemia (CLL) is the most common adult B-cell leukaemia and despite improvement in patients' outcome, following the use of targeted therapies, it remains incurable. CLL supportive microenvironment plays a key role in both CLL progression and drug resistance through signals that can be sensed by the main components of the focal adhesion complex, such as FAK and PYK2 kinases. Dysregulations of both kinases have been observed in several metastatic cancers, but their role in haematological malignancies is still poorly defined. We characterized FAK and PYK2 expression and observed that PYK2 expression is higher in leukaemic B cells and its overexpression significantly correlates with their malignant transformation. When targeting both FAK and PYK2 with the specific inhibitor defactinib, we observed a dose–response effect on CLL cells viability and survival. In vivo treatment of a CLL mouse model showed a decrease of the leukaemic clone in all the lymphoid organs along with a significant reduction of macrophages and of the spleen weight and size. Our results first define a possible prognostic value for PYK2 in CLL, and show that both FAK and PYK2 might become putative targets for both CLL and its microenvironment (e.g. macrophages), thus paving the way to an innovative therapeutic strategy.