Matriptase-2 inhibits HECV motility and tubule formation in vitro and tumour angiogenesis in vivo

Ezrin, primarily acts as a linker between the plasma membrane and the cytoskeleton, is involved in many cellular functions, including regulation of actin cytoskeleton, control of cell shape, adhesion, motility, and modulation of signaling pathways. Although ezrin is now recognized as a key component in tumor metastasis, its roles and the underlying mechanisms remain unclear. In the present study, we chose highly metastatic human lung carcinoma 95D cells, which highly express the ezrin proteins, as a model to examine the functional roles of ezrin in tumor suppression. An ezrin-silenced 95D cell line was established using lentivirus-mediated short hairpin RNA method. CCK-8 assay and soft agar assay analysis showed that downregulation of ezrin significantly suppressed the tumorigenicity and proliferation of 95D cells in vitro. cell migration and invasion studies showed that ezrin-specific deficiency in the cells caused the substantial reduction of the cell migration and invasion. In parallel, it also induced rearrangements of the actin cytoskeleton. Flow cytometry assay showed that changes in the ezrin protein level significantly affected the cell cycle distribution and eventual apoptosis. Furthermore, further studies showed that ezrin regulated the expression level of E-cadherin and CD44, which are key molecules involved in cell growth, migration, and invasion. Meanwhile, the suppression of ezrin expression also sensitized cells to antitumor drugs. Altogether, our results demonstrated that ezrin played an important role in the tumorigenicity and metastasis of lung cancer cells, which will benefit the development of therapeutic strategy for lung cancer.     

Ciliopathies with skeletal anomalies and renal insufficiency due to mutations in the IFT-A gene WDR19

A subset of ciliopathies, including Sensenbrenner, Jeune, and short-rib polydactyly syndromes are characterized by skeletal anomalies accompanied by multiorgan defects such as chronic renal failure and retinitis pigmentosa. Through exome sequencing we identified compound heterozygous mutations in WDR19 in a Norwegian family with Sensenbrenner syndrome. In a Dutch family with the clinically overlapping Jeune syndrome, a homozygous missense mutation in the same gene was found. Both families displayed a nephronophthisis-like nephropathy. Independently, we also identified compound heterozygous WDR19 mutations by exome sequencing in a Moroccan family with isolated nephronophthisis. WDR19 encodes IFT144, a member of the intraflagellar transport (IFT) complex A that drives retrograde ciliary transport. We show that IFT144 is absent from the cilia of fibroblasts from one of the Sensenbrenner patients and that ciliary abundance and morphology is perturbed, demonstrating the ciliary pathogenesis. Our results suggest that isolated nephronophthisis, Jeune, and Sensenbrenner syndromes are clinically overlapping disorders that can result from a similar molecular cause.     

Attenuation of scratch-induced reactive astrogliosis by novel EphA4 kinase inhibitors

The EphA4 receptor and its ephrin ligands are involved in astrocytic gliosis following CNS injury. Therefore, a strategy aimed at the blockade of EphA4 signaling could have broad therapeutic interest in brain disorders. We have identified novel small molecule inhibitors of EphA4 kinase in specific enzymatic and cell-based assays. In addition, we have demonstrated in two in vitro models of scratch injury that EphA4 receptor kinase is activated through phosphorylation and is involved in the repopulation of the wound after the scratch. A potent EphA4 kinase inhibitor significantly inhibited wound closure and reduced the accumulation of the reactive astrocytes inside the scratch. We have also shown that after the transient focal cerebral ischemia in rats, a large glial scar is formed by the accumulation of astrocytes and chondroitin sulfate proteoglycan surrounding the infarcted tissue at 7 days and 14 days of reperfusion. EphA4 protein expression is highly up-regulated in the same areas at these time points, supporting its potential role in the glial scar formation and maintenance. Taken together, these results suggest that EphA4 kinase inhibitors might interfere with the astrogliosis reaction and thereby lead to improved neurological outcome after ischemic injury.     

Assessment of the applicability of the Hertzian contact theory to edge-loaded prosthetic hip bearings

The components of prosthetic hip bearings may experience in-vivo subluxation and edge loading on the acetabular socket as a result of joint laxity, causing abnormally high, damaging contact stresses. In this research, edge-loaded contact of prosthetic hips is examined analytically and experimentally in the most commonly used categories of material pairs. In edge-loaded ceramic-on-ceramic hips, the Hertzian contact theory yields accurate (conservatively, <10% error) predictions of the contact dimensions. Moreover, the Hertzian theory successfully captures slope and curvature trends in the dependence of contact patch geometry on the applied load. In an edge-loaded ceramic-on-metal pair, a similar degree of accuracy is observed in the contact patch length; however, the contact width is less accurately predicted due to the onset of subsurface plasticity, which is predicted for loads >400N. The Hertzian contact theory is shown to be ill-suited to edge-loaded ceramic-on-polyethylene pairs due to polyethylene's nonlinear material behavior. This work elucidates the methods and the accuracy of applying classical contact theory to edge-loaded hip bearings. The results help to define the applicability of the Hertzian theory to the design of new components and materials to better resist severe edge loading contact stresses.     

Microbial production of bulk chemicals: development of anaerobic processes

Innovative fermentation processes are necessary for the cost-effective production of bulk chemicals from renewable resources. Current microbial processes are either anaerobic processes, with high yield and productivity, or less-efficient aerobic processes. Oxygen utilization plays an important role in energy generation and redox metabolism that is necessary for product formation. The aerobic productivity, however, is relatively low because of rate-limiting volumetric oxygen transfer; whereas the product yield in the presence of oxygen is generally low because part of the substrate is completely oxidized to CO?. Hence, new microbial conversion processes for the production of bulk chemicals should be anaerobic. In this opinion article, we describe different scenarios for the development of highly efficient microbial conversion processes for the anaerobic production of bulk chemicals.     

Structural basis for the Trembler-J phenotype of Charcot-Marie-Tooth disease

Mutations in peripheral myelin protein 22 (PMP22) can result in the common peripheral neuropathy Charcot-Marie-Tooth disease (CMTD). The Leu16Pro mutation in PMP22 results in misassembly of the protein, which causes the Trembler-J (TrJ) disease phenotype. Here we elucidate the structural defects present in a partially folded state of TrJ PMP22 that are decisive in promoting CMTD-causing misfolding. In this state, transmembrane helices 2-4 (TM2-4) form a molten globular bundle, while transmembrane helix 1 (TM1) is dissociated from this bundle. The TrJ mutation was seen to profoundly disrupt the TM1 helix, resulting in increased backbone dynamics and changes in the tertiary interactions of TM1 with the PMP22 TM2-4 core in the folded state. Consequently, TM1 undergoes enhanced dissociation from the other transmembrane segments in TrJ PMP22, becoming available for recognition and sequestration by protein-folding quality control, which leads to loss of function and toxic accumulation of aggregates that result in CMTD.     

High concentration electrophysiology-based fragment screen: discovery of novel acid-sensing ion channel 3 (ASIC3) inhibitors

The Merck Fragment Library was screened versus acid-sensing ion channel 3 (ASIC3), a novel target for the treatment of pain. Fragment hits were optimized using two strategies, and potency was improved from 0.7 mM to 3 μM with retention of good ligand efficiency and incorporation of reasonable physical properties, off-target profile, and rat pharmacokinetics.     

Towards an Absolute Chronology for the Aegean Iron Age: New Radiocarbon Dates from Lefkandi,Kalapodi and Corinth

The relative chronology of the Aegean Iron Age is robust. It is based on minute stylistic changes in the Submycenaean, Protogeometric and Geometric styles and their sub-phases. Yet, the absolute chronology of the time-span between the final stages of Late Helladic IIIC in the late second millennium BCE and the archaic colonization of Italy and Sicily toward the end of the 8^th century BCE lacks archaeological contexts that can be directly related to events carrying absolute dates mentioned in Egyptian/Near Eastern historical sources, or to well-dated Egyptian/Near Eastern rulers. The small number of radiocarbon dates available for this time span is not sufficient to establish an absolute chronological sequence. Here we present a new set of short-lived radiocarbon dates from the sites of Lefkandi, Kalapodi and Corinth in Greece. We focus on the crucial transition from the Submycenaean to the Protogeometric periods. This transition is placed in the late 11^th century BCE according to the Conventional Aegean Chronology and in the late 12^th century BCE according to the High Aegean Chronology. Our results place it in the second half of the 11^th century BCE.