Eph family receptor tyrosine kinases (including EphA3, EphB4) direct pathfinding of neurons within migratory fields of cells expressing gradients of their membrane-bound ligands. Others (EphB1 and ...EphA2) direct vascular network assembly, affecting endothelial migration, capillary morphogenesis, and angiogenesis. To explore how ephrins could provide positional labels for cell targeting, we tested whether endogenous endothelial and P19 cell EphB1 (ELK) and EphB2 (Nuk) receptors discriminate between different oligomeric forms of an ephrin-B1/Fc fusion ligand. Receptor tyrosine phosphorylation was stimulated by both dimeric and clustered multimeric ephrin-B1, yet only ephrin-B1 multimers (tetramers) promoted endothelial capillary-like assembly, cell attachment, and the recruitment of low-molecular-weight phosphotyrosine phosphatase (LMW-PTP) to receptor complexes. Cell-cell contact among cells expressing both EphB1 and ephrin-B1 was required for EphB1 activation and recruitment of LMW-PTP to EphB1 complexes. The EphB1-binding site for LMW-PTP was mapped and shown to be required for tetrameric ephrin-B1 to recruit LMW-PTP and to promote attachment. Thus, distinct EphB1-signaling complexes are assembled and different cellular attachment responses are determined by a receptor switch mechanism responsive to distinct ephrin-B1 oligomers.
The crystal structure of HPTP-B, a human isoenzyme of the low molecular weight phosphotyrosyl phosphatase (LMW PTPase) is reported here at a resolution of 1.6 Å. This high resolution structure of the ...second human LMW PTPase isoenzyme provides the opportunity to examine the structural basis of different substrate and inhibitor/activator responses. The crystal packing of HPTP-B positions a normally surface-exposed arginine in a position equivalent to the tyrosyl substrate. A comparison of all deposited crystallographic coordinates of these PTPases reveals three atomic positions within the active site cavity occupied by hydrogen bond donor or acceptor atoms on bound molecules, suggesting useful design elements for synthetic inhibitors. A selection of inhibitor and activator molecules as well as small molecule and peptide substrates were tested against each human isoenzyme. These results along with the crystal packing seen in HPTP-B suggest relevant sequence elements in the currently unknown target sequence.
Angiogenesis is a tightly controlled process in which signaling by the receptors for vascular endothelial growth factor (VEGF)
plays a key role. In order to define signaling pathways downstream of ...VEGF receptors (VEGFR), the kinase domain of VEGFR2
(Flk-1) was used as a bait to screen a human fetal heart library in the yeast two-hybrid system. One of the signaling molecules
identified in this effort was HCPTPA, a low molecular weight, cytoplasmic protein tyrosine phosphatase. Although HCPTPA possesses
no identifiable phosphotyrosine binding domains ( i.e. SH2 or phosphotyrosine binding domains), it bound specifically to active, autophosphorylated VEGFR2 but not to a mutated,
kinase-inactive VEGFR2. Recombinant VEGFR2 and endogenous VEGFR2 were substrates for recombinant HCPTPA, and HCPTPA was co-expressed
with VEGFR2 in endothelial cell lines, suggesting that HCPTPA may be a negative regulator of VEGFR2 signal transduction. To
pursue this possibility, an adenovirus directing the expression of HCPTPA was constructed. When used to infect cultured endothelial
cells, this adenovirus directed high level expression of HCPTPA that resulted in impairment of VEGF-mediated VEGFR2 autophosphorylation
and mitogen-activated protein kinase activation. Adenovirus-mediated overexpression of HCPTPA also inhibited VEGF-induced
cellular responses (endothelial cell migration and proliferation) and inhibited angiogenesis in the rat aortic ring assay.
Taken together, these findings indicate that HCPTPA may be an important regulator of VEGF-mediated signaling and biological
activity. Potential interactions with other signaling pathways and possible therapeutic implications are discussed.
The local hadronic calibration scheme developed for the reconstruction and calibration of jets and missing transverse energy in ATLAS has been evaluated using data obtained during combined beam tests ...of modules of the ATLAS liquid argon endcap and forward calorimeters. These tests covered the pseudorapidity range of 2.5 < | ? | < 4.0 . The analysis has been performed using special sets of calibration weights and corrections obtained with the Geant4 simulation of a detailed beam-test setup. The evaluation itself has been performed through the careful study of specific calorimeter performance parameters such as e.g. energy response and resolution, shower shapes, as well as different physics lists of the Geant4 simulation.
A full azimuthal
φ
-wedge of the ATLAS liquid argon end-cap calorimeter has been exposed to beams of electrons, muons and pions in the energy range
6
GeV
⩽
E
⩽
200
GeV
at the CERN SPS. The angular ...region studied corresponds to the ATLAS impact position around the pseudorapidity interval
1.6
<
|
η
|
<
1.8
. The beam test setup is described. A detailed study of the performance is given as well as the related intercalibration constants obtained. Following the ATLAS hadronic calibration proposal, a first study of the hadron calibration using a weighting ansatz is presented. The results are compared to predictions from Monte Carlo simulations, based on GEANT 3 and GEANT 4 models.
The local hadronic calibration scheme developed for the reconstruction and calibration of jets and missing transverse energy in ATLAS has been evaluated using data obtained during combined beam tests ...of modules of the ATLAS liquid argon endcap and forward calorimeters. These tests covered the pseudorapidity range of 2.5<|η|<4.0. The analysis has been performed using special sets of calibration weights and corrections obtained with the Geant4 simulation of a detailed beam-test setup. The evaluation itself has been performed through the careful study of specific calorimeter performance parameters such as e.g. energy response and resolution, shower shapes, as well as different physics lists of the Geant4 simulation.
Abstract Introduction Patients with unresectable liver tumors who fail initial treatment modalities have a poor prognosis (<1 yr). Although effective, delivery of high dose radiation therapy to these ...tumors is limited by proximity of radiosensitive bowel. We have previously reported that placement of a biologic mesh spacer (BMS) can effectively displace the bowel allowing for dose-intense radiation to be delivered with low short-term toxicity. The purpose of this study was to assess and report the long-term safety and oncologic outcomes of this cohort. Methods From 2012 to 2014 seven patients with unresectable hepatic malignancy (6 IHCC, 1 CRLM) underwent BMS (acellular human dermis) placement (2 open, 5 MIS) prior to radiation therapy. Prospective registry data were reviewed for tumor and treatment details, progression, metastasis and survival. RTOG guidelines were used to define radiation toxicities. Results Mean patient age was 50.4 years (30-62 years) and 4 patients were male (57.1%). Prior to surgery, all patients had been treated for an average of 12.5 months with surgery, chemotherapy, radiation and/or TACE. After surgery, all patients recovered well and received a mean radiation dose of 76.1 Gy (58.1-100 Gy) over 13 -25 fractions. 1 patient received SBRT; 4 fractions, 10 Gy each. Maximum dose delivered was 100 Gy (Biologic Equivalent Dose of 140 Gy, α/β =10). Mean time to initiation of radiation therapy was 24 days (12-48 days) from surgery. No significant GI toxicity was recorded, and no GI bleeding or ulcers were observed. Mean follow-up after XRT was 18.2 months (5.5 -31 months). Three patients had no loco-regional progression of disease. 2 patients had infield progression of liver disease and another had progressive lymphadenopathy. 3 patients developed pulmonary metastasis, at a mean time to distant failure of 3 months. There are 4 survivors over 2-years from surgery. Conclusion For patients with unresectable liver tumors, placement of a BMS enhances the safety and efficacy of high-dose radiotherapy, providing a survival benefit via delay in time to progression compared to traditional treatments with no significant short or long term GI toxicity.
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