Neuromonitoring und Neuroprotektion in der Kardioanästhesie: Bundesweite Umfrage des Arbeitskreises Kardioanästhesie der Deutschen Gesellschaft für Anästhesiologie und Intensivmedizin e.V

Zusammenfassung: Fragestellung: Primäres Ziel dieser in deutschen kardioanästhesiologischen Abteilungen durchgeführten Umfrage war, die aktuelle Praxis von Neuromonitoring und Neuroprotektion zu erheben. Methodik: Zwischen Oktober 2007 und Januar 2008 wurden hierzu Daten mithilfe eines 26Punkte umfassenden, anonymisierten Fragebogens erhoben. Ermittelt wurden: präoperative Evaluation der hirnversorgenden Arterien, intraoperatives Neuromonitoring, zerebroprotektive Maßnahmen, Perfusionsmanagement während extrakorporaler Zirkulation, postoperative Erhebung des neurologischen Status und Aus-/Weiterbildung im zerebralen Monitoring. Ergebnisse: 55% der Fragebögen wurden mit folgenden Angaben beantwortet: präoperative Duplexsonographie der Hirngefäße in 90% der Kliniken; intraoperatives Neuromonitoring mithilfe der Elektroenzephalographie (EEG; 60%) bei Typ-A-Dissektionen (38,1%), bei elektiven Operationen an der thorakalen bzw. thorakoabdominellen Aorta (34,1% resp. 31,6%) und in der Karotischirurgie (43,2%), weiterhin Einsatz der Nahinfrarotspektroskopie (40%), Ableitung evozierter Potenziale (30%) und transkranielle Dopplersonographie (17,5%). Auch kombinierte Verfahren wurden angewandt. Während Bypass-, Klappen und minimal-invasiven Operationen erfolgt meistens kein Monitoring des Zentralnervensystems. Zur Zerebroprotektion werden die Kühlung des Patienten an Herz-Lungen-Maschine (HLM; 100%), externe Kühlung des Kopfes (65%), Gabe von Kortikosteroiden (58%), Barbituraten (50%) und Antiepileptika (10%) eingesetzt. Als neuroprotektive Anästhesieverfahren gelten Inhalationsanästhesie (32,5%; Favorit: Sevofluran 76,5%) und total intravenöse Anästhesie (20%; Favoriten: Propofol und Barbiturate mit je 46,2%). Standardmäßig kühlen 72,5% der Krankenhäuser die Patienten bei Operationen mit Herz-Kreislauf-Stillstand, 37,5% bei allen Operationen mit HLM. Unter normothermen Bedingungen entspricht in 84,6% der Kliniken der HLM-Fluss dem errechneten Herzzeitvolumen (HZV), der anzustrebende mittlere arterielle Druck (MAP) liegt bei 60-70mmHg (43,9%) bzw. 50-60mmHg (41,5%). Bei einer Körpertemperatur unter 18°C wird der HLM-Fluss unter das errechnete HZV gesenkt (70%), während in 27% der Kliniken normotherme Flussraten gefahren werden. Der bevorzugte MAP unter Hypothermie liegt zwischen 50 und 60mmHg (59%). Intraoperatives Neuromonitoring wird im Narkoseprotokoll (77%) dokumentiert. Postoperativ wird der neurologische Status in 42,5% der Kliniken durch individuelle Einschätzung des Anästhesisten (77,5%) erhoben. Fortbildungen zum Thema Neuromonitoring werden in 32,5% der Kliniken regelmäßig organisiert, in 37,5% dem Arzt selbst überlassen. Schlussfolgerung: Das kardioanästhesiologische Vorgehen in Deutschland ist im Bereich Neuromonitoring und neuroprotektive Therapie nicht standardisiert. Ein "multimodales Neuromonitoring" wäre wünschenswer

Higgs Boson Masses in the Complex NMSSM at One-Loop Level

The Next-to-Minimal Supersymmetric Extension of the Standard Model (NMSSM) with a Higgs sector containing five neutral and two charged Higgs bosons allows for a rich phenomenology. In addition, the plethora of parameters provides many sources of CP violation. In contrast to the Minimal Supersymmetric Extension, CP violation in the Higgs sector is already possible at tree-level. For a reliable understanding and interpretation of the experimental results of the Higgs boson search, and for a proper distinction of Higgs sectors provided by the Standard Model or possible extensions, the Higgs boson masses have to be known as precisely as possible including higher-order corrections. In this paper we calculate the one-loop corrections to the neutral Higgs boson masses in the complex NMSSM in a Feynman diagrammatic approach adopting a mixed renormalization scheme based on on-shell and $\bar{DR}$ conditions. We study various scenarios where we allow for tree-level CP-violating phases in the Higgs sector and where we also study radiatively induced CP violation due to a non-vanishing phase of the trilinear coupling $A_t$ in the stop sector. The effects on the Higgs boson phenomenology are found to be significant. We furthermore estimate the theoretical error due to unknown higher-order corrections by both varying the renormalization scheme of the top and bottom quark masses and by adopting different renormalization scales. The residual theoretical error can be estimated to about 10%

A precision study of the fine tuning in the DiracNMSSM

Recently the DiracNMSSM has been proposed as a possible solution to reduce the fine tuning in supersymmetry. We determine the degree of fine tuning needed in the DiracNMSSM with and without non-universal gaugino masses and compare it with the fine tuning in the GNMSSM. To apply reasonable cuts on the allowed parameter regions we perform a precise calculation of the Higgs mass. In addition, we include the limits from direct SUSY searches and dark matter abundance. We find that both models are comparable in terms of fine tuning, with the minimal fine tuning in the GNMSSM slightly smaller.Comment: 20 pages + appendices, 10 figure

A high-content small molecule screen identifies sensitivity of glioblastoma stem cells to inhibition of polo-like kinase 1

Glioblastoma multiforme (GBM) is the most common primary brain cancer in adults and there are few effective treatments. GBMs contain cells with molecular and cellular characteristics of neural stem cells that drive tumour growth. Here we compare responses of human glioblastoma-derived neural stem (GNS) cells and genetically normal neural stem (NS) cells to a panel of 160 small molecule kinase inhibitors. We used live-cell imaging and high content image analysis tools and identified JNJ-10198409 (J101) as an agent that induces mitotic arrest at prometaphase in GNS cells but not NS cells. Antibody microarrays and kinase profiling suggested that J101 responses are triggered by suppression of the active phosphorylated form of polo-like kinase 1 (Plk1) (phospho T210), with resultant spindle defects and arrest at prometaphase. We found that potent and specific Plk1 inhibitors already in clinical development (BI 2536, BI 6727 and GSK 461364) phenocopied J101 and were selective against GNS cells. Using a porcine brain endothelial cell blood-brain barrier model we also observed that these compounds exhibited greater blood-brain barrier permeability in vitro than J101. Our analysis of mouse mutant NS cells (INK4a/ARF(-/-), or p53(-/-)), as well as the acute genetic deletion of p53 from a conditional p53 floxed NS cell line, suggests that the sensitivity of GNS cells to BI 2536 or J101 may be explained by the lack of a p53-mediated compensatory pathway. Together these data indicate that GBM stem cells are acutely susceptible to proliferative disruption by Plk1 inhibitors and that such agents may have immediate therapeutic value

Quantification of SLIT-ROBO transcripts in hepatocellular carcinoma reveals two groups of genes with coordinate expression

<p>Abstract</p> <p>Background</p> <p>SLIT-ROBO families of proteins mediate axon pathfinding and their expression is not solely confined to nervous system. Aberrant expression of <it>SLIT-ROBO </it>genes was repeatedly shown in a wide variety of cancers, yet data about their collective behavior in hepatocellular carcinoma (HCC) is missing. Hence, we quantified <it>SLIT-ROBO </it>transcripts in HCC cell lines, and in normal and tumor tissues from liver.</p> <p>Methods</p> <p>Expression of <it>SLIT-ROBO </it>family members was quantified by real-time qRT-PCR in 14 HCC cell lines, 8 normal and 35 tumor tissues from the liver. ANOVA and Pearson's correlation analyses were performed in R environment, and different clinicopathological subgroups were pairwise compared in Minitab. Gene expression matrices of cell lines and tissues were analyzed by Mantel's association test.</p> <p>Results</p> <p>Genewise hierarchical clustering revealed two subgroups with coordinate expression pattern in both the HCC cell lines and tissues: <it>ROBO1</it>, <it>ROBO2</it>, <it>SLIT1 </it>in one cluster, and <it>ROBO4</it>, <it>SLIT2</it>, <it>SLIT3 </it>in the other, respectively. Moreover, <it>SLIT-ROBO </it>expression predicted <it>AFP</it>-dependent subgrouping of HCC cell lines, but not that of liver tissues. <it>ROBO1 </it>and <it>ROBO2 </it>were significantly up-regulated, whereas <it>SLIT3 </it>was significantly down-regulated in cell lines with high-<it>AFP </it>background. When compared to normal liver tissue, <it>ROBO1 </it>was found to be significantly overexpressed, while <it>ROBO4 </it>was down-regulated in HCC. We also observed that <it>ROBO1 </it>and <it>SLIT2 </it>differentiated histopathological subgroups of liver tissues depending on both tumor staging and differentiation status. However, <it>ROBO4 </it>could discriminate poorly differentiated HCC from other subgroups.</p> <p>Conclusion</p> <p>The present study is the first in comprehensive and quantitative evaluation of <it>SLIT-ROBO </it>family gene expression in HCC, and suggests that the expression of <it>SLIT-ROBO </it>genes is regulated in hepatocarcinogenesis. Our results implicate that <it>SLIT-ROBO </it>transcription profile is bi-modular in nature, and that each module shows intrinsic variability. We also provide quantitative evidence for potential use of <it>ROBO1</it>, <it>ROBO4 </it>and <it>SLIT2 </it>for prediction of tumor stage and differentiation status.</p

AGO6 Functions in RNA-Mediated Transcriptional Gene Silencing in Shoot and Root Meristems in Arabidopsis thaliana

RNA-directed DNA methylation (RdDM) is a small interfering RNA (siRNA)-mediated epigenetic modification that contributes to transposon silencing in plants. RdDM requires a complex transcriptional machinery that includes specialized RNA polymerases, named Pol IV and Pol V, as well as chromatin remodelling proteins, transcription factors, RNA binding proteins, and other plant-specific proteins whose functions are not yet clarified. In Arabidopsis thaliana, DICER-LIKE3 and members of the ARGONAUTE4 group of ARGONAUTE (AGO) proteins are involved, respectively, in generating and using 24-nt siRNAs that trigger methylation and transcriptional gene silencing of homologous promoter sequences. AGO4 is the main AGO protein implicated in the RdDM pathway. Here we report the identification of the related AGO6 in a forward genetic screen for mutants defective in RdDM and transcriptional gene silencing in shoot and root apical meristems in Arabidopsis thaliana. The identification of AGO6, and not AGO4, in our screen is consistent with the primary expression of AGO6 in shoot and root growing points

Growth arrest-specific transcript 5 associated snoRNA levels are related to p53 expression and DNA damage in colorectal cancer

BACKGROUND The growth arrest-specific transcript 5 gene (GAS5) encodes a long noncoding RNA (lncRNA) and hosts a number of small nucleolar RNAs (snoRNAs) that have recently been implicated in multiple cellular processes and cancer. Here, we investigate the relationship between DNA damage, p53, and the GAS5 snoRNAs to gain further insight into the potential role of this locus in cell survival and oncogenesis both in vivo and in vitro. METHODS We used quantitative techniques to analyse the effect of DNA damage on GAS5 snoRNA expression and to assess the relationship between p53 and the GAS5 snoRNAs in cancer cell lines and in normal, pre-malignant, and malignant human colorectal tissue and used biological techniques to suggest potential roles for these snoRNAs in the DNA damage response. RESULTS GAS5-derived snoRNA expression was induced by DNA damage in a p53-dependent manner in colorectal cancer cell lines and their levels were not affected by DICER. Furthermore, p53 levels strongly correlated with GAS5-derived snoRNA expression in colorectal tissue. CONCLUSIONS In aggregate, these data suggest that the GAS5-derived snoRNAs are under control of p53 and that they have an important role in mediating the p53 response to DNA damage, which may not relate to their function in the ribosome. We suggest that these snoRNAs are not processed by DICER to form smaller snoRNA-derived RNAs with microRNA (miRNA)-like functions, but their precise role requires further evaluation. Furthermore, since GAS5 host snoRNAs are often used as endogenous controls in qPCR quantifications we show that their use as housekeeping genes in DNA damage experiments can lead to inaccurate results

MicroRNA-96 Directly Inhibits γ-Globin Expression in Human Erythropoiesis

Fetal hemoglobin, HbF (α2γ2), is the main hemoglobin synthesized up to birth, but it subsequently declines and adult hemoglobin, HbA (α2β2), becomes predominant. Several studies have indicated that expression of the HbF subunit γ-globin might be regulated post-transcriptionally. This could be confered by ∼22-nucleotide long microRNAs that associate with argonaute proteins to specifically target γ-globin mRNAs and inhibit protein expression. Indeed, applying immunopurifications, we found that γ-globin mRNA was associated with argonaute 2 isolated from reticulocytes that contain low levels of HbF (<1%), whereas association was significantly lower in reticulocytes with high levels of HbF (90%). Comparing microRNA expression in reticulocytes from cord blood and adult blood, we identified several miRNAs that were preferentially expressed in adults, among them miRNA-96. The overexpression of microRNA-96 in human ex vivo erythropoiesis decreased γ-globin expression by 50%, whereas the knock-down of endogenous microRNA-96 increased γ-globin expression by 20%. Moreover, luciferase reporter assays showed that microRNA-96 negatively regulates expression of γ-globin in HEK293 cells, which depends on a seedless but highly complementary target site located within the coding sequence of γ-globin. Based on these results we conclude that microRNA-96 directly suppresses γ-globin expression and thus contributes to HbF regulation

TRAIL Death Receptor-4, Decoy Receptor-1 and Decoy Receptor-2 Expression on CD8+ T Cells Correlate with the Disease Severity in Patients with Rheumatoid Arthritis

BACKGROUND: Rheumatoid Arthritis (RA) is a chronic autoimmune inflammatory disorder. Although the pathogenesis of disease is unclear, it is well known that T cells play a major role in both development and perpetuation of RA through activating macrophages and B cells. Since the lack of TNF-Related Apoptosis Inducing Ligand (TRAIL) expression resulted in defective thymocyte apoptosis leading to an autoimmune disease, we explored evidence for alterations in TRAIL/TRAIL receptor expression on peripheral T lymphocytes in the molecular mechanism of RA development. METHODS: The expression of TRAIL/TRAIL receptors on T cells in 20 RA patients and 12 control individuals were analyzed using flow cytometry. The correlation of TRAIL and its receptor expression profile was compared with clinical RA parameters (RA activity scored as per DAS28) using Spearman Rho Analysis. RESULTS: While no change was detected in the ratio of CD4+ to CD8+ T cells between controls and RA patient groups, upregulation of TRAIL and its receptors (both death and decoy) was detected on both CD4+ and CD8+ T cells in RA patients compared to control individuals. Death Receptor-4 (DR4) and the decoy receptors DcR1 and DcR2 on CD8+ T cells, but not on CD4+ T cells, were positively correlated with patients' DAS scores. CONCLUSIONS: Our data suggest that TRAIL/TRAIL receptor expression profiles on T cells might be important in revelation of RA pathogenesis