ALBERT

Beschreibung: Ras GTPase-activating proteins (GAPs) are important regulators for Ras activation, which is instrumental in tumor development. However, the mechanism underlying this regulation remains elusive. We demonstrate here that activated EGFR phosphorylates the Y593 residue of the protein known as family with sequence similarity 129, member B (FAM129B), which is overexpressed in many types of human cancer. FAM129B phosphorylation increased the interaction between FAM129B and Ras, resulting in reduced binding of p120-RasGAP to Ras. FAM129B phosphorylation promoted Ras activation, increasing ERK1/2- and PKM2-dependent β-catenin transactivation and leading to the enhanced glycolytic gene expression and the Warburg effect; promoting tumor cell proliferation and invasion; and supporting brain tumorigenesis. Our studies unearthed a novel and important mechanism underlying EGFR-mediated Ras activation in tumor development.

Beschreibung: Single-span membrane proteins (ssMPs) represent approximately one-half of all membrane proteins and play important roles in cellular communications. However, like all membrane proteins, ssMPs are prone to misfolding and aggregation because of the hydrophobicity of transmembrane helices, making them difficult to study using common aqueous solution-based approaches. Detergents and membrane mimetics can solubilize membrane proteins but do not always result in proper folding and functionality. Here, we use cell-free protein synthesis in the presence of oil drops to create a one-pot system for the synthesis, assembly, and display of functional ssMPs. Our studies suggest that oil drops prevent aggregation of some in vitro-synthesized ssMPs by allowing these ssMPs to localize on oil surfaces. We speculate that oil drops may provide a hydrophobic interior for cotranslational insertion of the transmembrane helices and a fluidic surface for proper assembly and display of the ectodomains. These functionalized oil drop surfaces could mimic cell surfaces and allow ssMPs to interact with cell surface receptors under an environment closest to cell–cell communication. Using this approach, we showed that apoptosis-inducing human transmembrane proteins, FasL and TRAIL, synthesized and displayed on oil drops induce apoptosis of cultured tumor cells. In addition, we take advantage of hydrophobic interactions of transmembrane helices to manipulate the assembly of ssMPs and create artificial clusters on oil drop surfaces. Thus, by coupling protein synthesis with self-assembly at the water–oil interface, we create a platform that can use recombinant ssMPs to communicate with cells.

Beschreibung: Wetting of actual surfaces involves diverse hysteretic phenomena stemming from ever-present imperfections. Here, we clarify the origin of wetting hysteresis for a liquid front advancing or receding across an isolated defect of nanometric size. Various kinds of chemical and topographical nanodefects, which represent salient features of actual heterogeneous surfaces, are investigated. The most probable wetting path across surface heterogeneities is identified by combining, within an innovative approach, microscopic classical density functional theory and the string method devised for the study of rare events. The computed rugged free-energy landscape demonstrates that hysteresis emerges as a consequence of metastable pinning of the liquid front at the defects; the barriers for thermally activated defect crossing, the pinning force, and hysteresis are quantified and related to the geometry and chemistry of the defects allowing for the occurrence of nanoscopic effects. The main result of our calculations is that even weak nanoscale defects, which are difficult to characterize in generic microfluidic experiments, can be the source of a plethora of hysteretical phenomena, including the pinning of nanobubbles.

Beschreibung: Pathologic extraskeletal bone formation, or heterotopic ossification (HO), occurs following mechanical trauma, burns, orthopedic operations, and in patients with hyperactivating mutations of the type I bone morphogenetic protein receptor ACVR1 (Activin type 1 receptor). Extraskeletal bone forms through an endochondral process with a cartilage intermediary prompting the hypothesis that hypoxic signaling present during cartilage formation drives HO development and that HO precursor cells derive from a mesenchymal lineage as defined by Paired related homeobox 1 (Prx). Here we demonstrate that Hypoxia inducible factor-1α (Hif1α), a key mediator of cellular adaptation to hypoxia, is highly expressed and active in three separate mouse models: trauma-induced, genetic, and a hybrid model of genetic and trauma-induced HO. In each of these models, Hif1α expression coincides with the expression of master transcription factor of cartilage, Sox9 [(sex determining region Y)-box 9]. Pharmacologic inhibition of Hif1α using PX-478 or rapamycin significantly decreased or inhibited extraskeletal bone formation. Importantly, de novo soft-tissue HO was eliminated or significantly diminished in treated mice. Lineage-tracing mice demonstrate that cells forming HO belong to the Prx lineage. Burn/tenotomy performed in lineage-specific Hif1α knockout mice (Prx-Cre/Hif1αfl:fl) resulted in substantially decreased HO, and again lack of de novo soft-tissue HO. Genetic loss of Hif1α in mesenchymal cells marked by Prx-cre prevents the formation of the mesenchymal condensations as shown by routine histology and immunostaining for Sox9 and PDGFRα. Pharmacologic inhibition of Hif1α had a similar effect on mesenchymal condensation development. Our findings indicate that Hif1α represents a promising target to prevent and treat pathologic extraskeletal bone.

Beschreibung: Cilia (eukaryotic flagella) are present in diverse eukaryotic lineages and have essential motility and sensory functions. The cilium’s capacity to sense and transduce extracellular signals depends on dynamic trafficking of ciliary membrane proteins. This trafficking is often mediated by the Bardet–Biedl Syndrome complex (BBSome), a protein complex for which the precise subcellular distribution and mechanisms of action are unclear. In humans, BBSome defects perturb ciliary membrane protein distribution and manifest clinically as Bardet–Biedl Syndrome. Cilia are also important in several parasites that cause tremendous human suffering worldwide, yet biology of the parasite BBSome remains largely unexplored. We examined BBSome functions in Trypanosoma brucei, a flagellated protozoan parasite that causes African sleeping sickness in humans. We report that T. brucei BBS proteins assemble into a BBSome that interacts with clathrin and is localized to membranes of the flagellar pocket and adjacent cytoplasmic vesicles. Using BBS gene knockouts and a mouse infection model, we show the T. brucei BBSome is dispensable for flagellar assembly, motility, bulk endocytosis, and cell viability but required for parasite virulence. Quantitative proteomics reveal alterations in the parasite surface proteome of BBSome mutants, suggesting that virulence defects are caused by failure to maintain fidelity of the host–parasite interface. Interestingly, among proteins altered are those with ubiquitination-dependent localization, and we find that the BBSome interacts with ubiquitin. Collectively, our data indicate that the BBSome facilitates endocytic sorting of select membrane proteins at the base of the cilium, illuminating BBSome roles at a critical host–pathogen interface and offering insights into BBSome molecular mechanisms.

Beschreibung: Publication date: Available online 29 December 2015 Source: Geodesy and Geodynamics Author(s): Fuqiong Huang, Yong Chen, Ping Ji, Kexin Ren, Fuwang Gao, Lingkong Zhang The Kunlunshan Mountain M s8.1 earthquake, occurred in Nov.14, 2001, is the first event with magnitude more than 8 in the China earthquake monitoring history, specifically at the beginning of digital techniques in precursor monitoring networks. Any investigation of recorded data on this earthquake is very important for testing the operation of the digital monitoring networks and understanding the preparation, occurrence, and adjustment of stress/strain of strong continental earthquakes. In this paper we investigated the co-seismic response changes of well water level of groundwater and volume strain meter of bore hole in digital earthquake monitoring network of Capital area and its vicinity, due to the Nov.14, 2001 M s8.1 Kunlun Mountain earthquake. The responding time, shapes or manners, amplitudes, and lasting time of well water level and strain-meters to seismic wave are studied in comparison. Then we discussed the possibility that the response changes of groundwater to strong distant earthquakes can be understood as one kind of observing evidence of stress/strain changes induced by distant earthquake.

Beschreibung: A long-term study of O, H and C stable isotopes has been undertaken on river waters across the 7000 km 2 upper Thames lowland river basin in the southern UK. During the period, flow conditions ranged from drought to flood. A 10-year monthly record (2003–2012) of the main River Thames showed a maximum variation of 3‰ (δ 18 O) and 20‰ (δ 2 H), though inter-annual average values varied little around a mean of –6.5‰ (δ 18 O) and –44‰ (δ 2 H). A δ 2 H/δ 18 O slope of 5.3 suggested a degree of evaporative enrichment, consistent with derivation from local rainfall with a weighted mean of –7.2‰ (δ 18 O) and –48‰ (δ 2 H) for the period. A tendency towards isotopic depletion of the river with increasing flowrate was noted, but at very high flows (〉100 m 3 /s) a reversion to the mean was interpreted as the displacement of bank storage by rising groundwater levels (corroborated by measurements of specific electrical conductivity). A shorter quarterly study (October 2011 – April 2013) of isotope variations in 15 tributaries with varying geology revealed different responses to evaporation, with a well-correlated inverse relationship between Δ 18 O and baseflow index (BFI) for most of the rivers. A comparison with aquifer waters in the basin showed that even at low flow, rivers rarely consist solely of isotopically unmodified groundwater. Long-term monitoring (2003–2007) of carbon stable isotopes in dissolved inorganic carbon (DIC) in the Thames revealed a complex interplay between respiration, photosynthesis and evasion, but with a mean inter-annual δ 13 C-DIC value of –14.8 ± 0.5‰, exchange with atmospheric carbon could be ruled out. Quarterly monitoring of the tributaries (October 2011 – April 2013) indicated that in addition to the above factors, river flow variations and catchment characteristics were likely to affect δ 13 C-DIC. Comparison with basin groundwaters of different alkalinity and δ 13 C-DIC values showed that the origin of river baseflow is usually obscured. The findings show that long-term monitoring of environmental tracers can help to improve the understanding of how lowland river catchments function. This article is protected by copyright. All rights reserved.

Beschreibung: Publication date: Available online 29 December 2015 Source: Journal of Rock Mechanics and Geotechnical Engineering Author(s): Yankun Sun, Qi Li, Duoxing Yang, Xuehao Liu Carbon dioxide (CO 2 ) geosequestration in deep saline aquifers has been currently deemed as a preferable and practicable mitigation means for reducing anthropogenic greenhouse gases (GHGs) emissions to the atmosphere, as deep saline aquifers can offer the greatest potential from a capacity point of view. Hence, research on core-scale CO 2 /brine multiphase migration processes is of great significance for precisely estimating storage efficiency, ensuring storage security, and predicting the long-term effects of the sequestered CO 2 in subsurface saline aquifers. This review article initially presents a brief description of the essential aspects of CO 2 subsurface transport and geological trapping mechanisms, and then outlines the state-of-the-art laboratory core flooding experimental apparatus that has been adopted for simulating CO 2 injection and migration processes in the literature over the past decade. Finally, a summary of the characteristics, components and applications of publicly reported core flooding equipment as well as major research gaps and areas in need of further study are given in relevance to laboratory-scale core flooding experiments in CO 2 geosequestration under reservoir conditions.

Beschreibung: UK flood policy needs a complete overhaul says the Environment Agency, including accepting that flooding is inevitable