Blockade of the extracellular signal-regulated kinase pathway by U0126 attenuates neuronal damage following circulatory arrest

AbstractObjectivesThe extracellular signal-regulated kinase pathway of the mitogen-activated protein kinase signal transduction cascade has been implicated in the neuronal and endothelial dysfunction witnessed following cerebral ischemia-reperfusion injury. Extracellular signal-regulated kinase is activated by mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2. We evaluated the ability of a mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2–specific inhibitor (U0126) to block extracellular signal-regulated kinase activation and mitigate ischemic neuronal damage in a model of deep hypothermic circulatory arrest.MethodsPiglets underwent normal flow cardiopulmonary bypass (control, n = 4), deep hypothermic circulatory arrest (n = 6), and deep hypothermic circulatory arrest with U0126 (n = 5) at 20°C for 60 minutes. The deep hypothermic circulatory arrest with U0126 group was given 200 μg/kg of U0126 45 minutes prior to initiation of bypass followed by 100 μg/kg at reperfusion. Following 24 hours of post–cardiopulmonary bypass recovery, brains were harvested. Eleven distinct cortical regions were evaluated for neuronal damage using hematoxylin and eosin staining. A section of ischemic cortex was further evaluated by immunohistochemistry with rabbit polyclonal antibody against phosphorylated extracellular signal-regulated kinase 1/2.ResultsThe deep hypothermic circulatory arrest and deep hypothermic circulatory arrest with U0126 groups displayed diffuse ischemic changes. However, the deep hypothermic circulatory arrest with U0126 group possessed significantly lower neuronal damage scores in the right frontal watershed zone of cerebral cortex, basal ganglia, and thalamus (P ≤ .05) and an overall trend toward neuroprotection versus the deep hypothermic circulatory arrest group. This neuroprotection was accompanied by nearly complete blockade of phosphorylated extracellular signal-regulated kinase in the cerebral vascular endothelium.ConclusionsIn this experimental model of deep hypothermic circulatory arrest, U0126 blocked extracellular signal-regulated kinase activation and provided a significant neuroprotective effect. These results support targeting of the extracellular signal-regulated kinase pathway for inhibition as a novel therapeutic approach to mitigate neuronal damage following deep hypothermic circulatory arrest

Spin Correlation in tt-bar Production from pp-bar Collisions at sqrt(s)=1.8 TeV

The D0 collaboration has performed a study of spin correlation in tt-bar production for the process tt-bar to bb-bar W^+W^-, where the W bosons decay to e-nu or mu-nu. A sample of six events was collected during an exposure of the D0 detector to an integrated luminosity of approximately 125 pb^-1 of sqrt{s}=1.8 TeV pp-bar collisions. The standard model (SM) predicts that the short lifetime of the top quark ensures the transmission of any spin information at production to the tt-bar decay products. The degree of spin correlation is characterized by a correlation coefficient k. We find that k>-0.25 at the 68% confidence level, in agreement with the SM prediction of k=0.88.Comment: Submitted to PRL, Added references, minor changes to tex

Characterization of TEM1/endosialin in human and murine brain tumors

<p>Abstract</p> <p>Background</p> <p><it>TEM1/endosialin </it>is an emerging microvascular marker of tumor angiogenesis. We characterized the expression pattern of <it>TEM1/endosialin </it>in astrocytic and metastatic brain tumors and investigated its role as a therapeutic target in human endothelial cells and mouse xenograft models.</p> <p>Methods</p> <p><it>In situ </it>hybridization (ISH), immunohistochemistry (IH) and immunofluorescence (IF) were used to localize <it>TEM1/endosialin </it>expression in grade II-IV astrocytomas and metastatic brain tumors on tissue microarrays. Changes in <it>TEM1/endosialin </it>expression in response to pro-angiogenic conditions were assessed in human endothelial cells grown <it>in vitro</it>. Intracranial U87MG glioblastoma (GBM) xenografts were analyzed in nude <it>TEM1/endosialin </it>knockout (KO) and wildtype (WT) mice.</p> <p>Results</p> <p><it>TEM1/endosialin </it>was upregulated in primary and metastatic human brain tumors, where it localized primarily to the tumor vasculature and a subset of tumor stromal cells. Analysis of 275 arrayed grade II-IV astrocytomas demonstrated <it>TEM1/endosialin </it>expression in 79% of tumors. Robust <it>TEM1/endosialin </it>expression occurred in 31% of glioblastomas (grade IV astroctyomas). <it>TEM1/endosialin </it>expression was inversely correlated with patient age. TEM1/endosialin showed limited co-localization with CD31, αSMA and fibronectin in clinical specimens. <it>In vitro</it>, <it>TEM1/endosialin </it>was upregulated in human endothelial cells cultured in matrigel. Vascular <it>Tem1/endosialin </it>was induced in intracranial U87MG GBM xenografts grown in mice. <it>Tem1/endosialin </it>KO vs WT mice demonstrated equivalent survival and tumor growth when implanted with intracranial GBM xenografts, although <it>Tem1/endosialin </it>KO tumors were significantly more vascular than the WT counterparts.</p> <p>Conclusion</p> <p><it>TEM1/endosialin </it>was induced in the vasculature of high-grade brain tumors where its expression was inversely correlated with patient age. Although lack of <it>TEM1/endosialin </it>did not suppress growth of intracranial GBM xenografts, it did increase tumor vascularity. The cellular localization of <it>TEM1/endosialin </it>and its expression profile in primary and metastatic brain tumors support efforts to therapeutically target this protein, potentially via antibody mediated drug delivery strategies.</p

Search for mSUGRA in single-electron events with jets and large missing transverse energy in ppbar collisions at center of mass energy of 1.8 TeV

We describe a search for evidence of minimal supergravity (mSUGRA) in 92.7 pb^(-1) of data collected with the D{\O}detector at the Fermilab Tevatron ppbar collider at center of mass energy of 1.8 TeV. Events with a single electron, four or more jets, and large missing transverse energy were used in this search. The major backgrounds are from W+jets, misidentified multijet, ttbar, and WW production. We observe no excess above the expected number of background events in our data. A new limit in terms of mSUGRA model parameters is obtained.Comment: 24 pages, 17 figures; submitted to PRD; FERMILAB-Pub-02/074-

Helicity of the W Boson in Lepton+Jets ttbar Events

We examine properties of ttbar candidates events in lepton+jets final states to establish the helicities of the W bosons in t->W+b decays. Our analysis is based on a direct calculation of a probability that each event corresponds to a ttbar final state, as a function of the helicity of the W boson. We use the 125 events/pb sample of data collected by the DO experiment during Run I of the Fermilab Tevatron collider at sqrt{s}=1.8 TeV, and obtain a longitudinal helicity fraction of F_0=0.56+/-0.31, which is consistent with the prediction of F_0=0.70 from the standard model

Model-independent measurement of t\boldsymbol{t}-channel single top quark production in ppˉ\boldsymbol{p\bar{p}} collisions at s=1.96\boldsymbol{\sqrt{s}=1.96} TeV

We present a model-independent measurement of $t$-channel electroweak production of single top quarks in \ppbar collisions at $\sqrt{s}=1.96\;\rm TeV$. Using $5.4\;\rm fb^{-1}$ of integrated luminosity collected by the D0 detector at the Fermilab Tevatron Collider, and selecting events containing an isolated electron or muon, missing transverse energy and one or two jets originating from the fragmentation of $b$ quarks, we measure a cross section \sigma({\ppbar}{\rargap}tqb+X) = 2.90 \pm 0.59\;\rm (stat+syst)\; pb for a top quark mass of $172.5\;\rm GeV$. The probability of the background to fluctuate and produce a signal as large as the one observed is $1.6\times10^{-8}$, corresponding to a significance of 5.5 standard deviations.Comment: 8 pages, 4 figures, submitted to Phys. Lett.

Search for single top quark production in ppbar collisions at sqrt(s)=1.96 TeV

We present a search for electroweak production of single top quarks in the s-channel and t-channel using neural networks for signal-background separation. We have analyzed 230 pb$^{-1}$ of data collected with the D0 detector at the Fermilab Tevatron Collider at a center-of-mass energy of 1.96 TeV and find no evidence for a single top quark signal. The resulting 95% confidence level upper limits on the single top quark production cross sections are 6.4 pb in the s-channel and 5.0 pb in the t-channel.Comment: 9 pages, 4 figure

Mesothelin Expression in the Leptomeninges and Meningiomas

The identity and functions of surface proteins on human leptomeningeal and meningioma cells are incompletely characterized. Some structural and functional similarities between the leptomeninges and pleura suggest that proteins important to pleural function and tumorigenesis might also be relevant to leptomeningeal disease. Mesothelin is a recently described, 40-kDa membrane protein expressed in pleura. Its functions in this tissue are under investigation. Sections of 20 normal adult brains with leptomeninges and 49 World Health Organization (WHO) grade I, 21 grade II, and 2 grade III meningiomas were analyzed using an extensively characterized monoclonal antibody to mesothelin and streptavidin-biotin complex immunohistochemistry. Five meningiomas were also evaluated by Western blot. Mesothelin immunoreactivity was detected in the arachnoid in 6 of 20 cases and in 23 of 49 WHO grade I meningiomas. It was also detected in 7 of 21 WHO II tumors and 1 of the 2 anaplastic meningiomas. By Western blot, all five meningiomas exhibited mesothelin precursor protein, including one where notable immunoreactivity was not identified in a formalin-fixed tissue section. These findings suggest that mesothelin is expressed in at least some arachnoid and meningioma cells. Future studies may clarify its role in the development of meningiomas, meningeal seeding of gliomas, and metastases to the leptomeninges. (J Histochem Cytochem 56:579–585, 2008