TY - JOUR A1 - Koch, Oliver A1 - Cappel, Daniel A1 - Nocker, Monika A1 - Jaeger, Timo A1 - Flohé, Leopold A1 - Sotriffer, Christoph A1 - Selzer, Paul T1 - Virtual screening using structure-based consensus pharmacophore models and ensemble docking based on MD-generated conformations : [From 6th German Conference on Chemoinformatics, GCC 2010, Goslar, Germany. 7-9 November 2010] JF - Journal of Cheminformatics N2 - No abstract available. KW - chemistry Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142830 VL - 3 IS - Suppl. 1 ER - TY - JOUR A1 - Allignol, Arthur A1 - Schumacher, Martin A1 - Wanner, Christoph A1 - Drechsler, Christiane A1 - Beyersmann, Jan T1 - Understanding competing risks: a simulation point of view JF - BMC Medical Research Methodology N2 - Background: Competing risks methodology allows for an event-specific analysis of the single components of composite time-to-event endpoints. A key feature of competing risks is that there are as many hazards as there are competing risks. This is not always well accounted for in the applied literature. Methods: We advocate a simulation point of view for understanding competing risks. The hazards are envisaged as momentary event forces. They jointly determine the event time. Their relative magnitude determines the event type. 'Empirical simulations' using data from a recent study on cardiovascular events in diabetes patients illustrate subsequent interpretation. The method avoids concerns on identifiability and plausibility known from the latent failure time approach. Results: The 'empirical simulations' served as a proof of concept. Additionally manipulating baseline hazards and treatment effects illustrated both scenarios that require greater care for interpretation and how the simulation point of view aids the interpretation. The simulation algorithm applied to real data also provides for a general tool for study planning. Conclusions: There are as many hazards as there are competing risks. All of them should be analysed. This includes estimation of baseline hazards. Study planning must equally account for these aspects. KW - Cumulative incidence function KW - Clinical-trials KW - Sample-sizes KW - Regression KW - Subdistribution KW - Hazards KW - Model KW - Probabilities KW - Tests Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142811 VL - 11 IS - 86 ER - TY - JOUR A1 - Binder, Andreas A1 - May, Denisa A1 - Baron, Ralf A1 - Maier, Christoph A1 - Tölle, Thomas R. A1 - Treede, Rolf-Detlef A1 - Berthele, Achim A1 - Faltraco, Frank A1 - Flor, Herta A1 - Gierthmühlen, Janne A1 - Haenisch, Sierk A1 - Huge, Volker A1 - Magerl, Walter A1 - Maihöfner, Christian A1 - Richter, Helmut A1 - Rolke, Roman A1 - Scherens, Andrea A1 - Üçeyler, Nurcan A1 - Ufer, Mike A1 - Wasner, Gunnar A1 - Zhu, Jihong A1 - Cascorbi, Ingolf T1 - Transient Receptor Potential Channel Polymorphisms Are Associated with the Somatosensory Function in Neuropathic Pain Patients JF - PLoS ONE N2 - Transient receptor potential channels are important mediators of thermal and mechanical stimuli and play an important role in neuropathic pain. The contribution of hereditary variants in the genes of transient receptor potential channels to neuropathic pain is unknown. We investigated the frequency of transient receptor potential ankyrin 1, transient receptor potential melastin 8 and transient receptor potential vanilloid 1 single nucleotide polymorphisms and their impact on somatosensory abnormalities in neuropathic pain patients. Within the German Research Network on Neuropathic Pain (Deutscher Forscbungsverbund Neuropathischer Schmerz) 371 neuropathic pain patients were phenotypically characterized using standardized quantitative sensory testing. Pyrosequencing was employed to determine a total of eleven single nucleotide polymorphisms in transient receptor potential channel genes of the neuropathic pain patients and a cohort of 253 German healthy volunteers. Associations of quantitative sensory testing parameters and single nucleotide polymorphisms between and within groups and subgroups, based on sensory phenotypes, were analyzed. Single nucleotide polymorphisms frequencies did not differ between both the cohorts. However, in neuropathic pain patients transient receptor potential ankyrin 1 710G>A (rs920829, E179K) was associated with the presence of paradoxical heat sensation (p=0.03), and transient receptor potential vanilloid 1 1911A>G (rs8065080, I585V) with cold hypoalgesia (p=0.0035). Two main subgroups characterized by preserved (1) and impaired (2) sensory function were identified. In subgroup 1 transient receptor potential vanilloid 1 1911A>G led to significantly less heat hyperalgesia, pinprick hyperalgesia and mechanical hypaesthesia (p=0.006, p=0.005 and p<0.001) and transient receptor potential vanilloid 1 1103C>G (rs222747, M315I) to cold hypaesthesia (p=0.002), but there was absence of associations in subgroup 2. In this study we found no evidence that genetic variants of transient receptor potential channels are involved in the expression of neuropathic pain, but transient receptor potential channel polymorphisms contributed significantly to the somatosensory abnormalities of neuropathic pain patients. KW - Paradoxical heat sensation KW - Neurogenic inflammation KW - Capsaicin receptor KW - TRP Channels KW - Cold KW - Mechanisms KW - Hyperalgesia KW - Sensitivity KW - Expression KW - Stimuli Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142782 VL - 6 IS - 3 ER - TY - JOUR A1 - Arhondakis, Stilianos A1 - Frousios, Kimon A1 - Iliopoulos, Costas S. A1 - Pissis, Solon P. A1 - Tischler, German A1 - Kossida, Sophia T1 - Transcriptome map of mouse isochores JF - BMC Genomics N2 - Background: The availability of fully sequenced genomes and the implementation of transcriptome technologies have increased the studies investigating the expression profiles for a variety of tissues, conditions, and species. In this study, using RNA-seq data for three distinct tissues (brain, liver, and muscle), we investigate how base composition affects mammalian gene expression, an issue of prime practical and evolutionary interest. Results: We present the transcriptome map of the mouse isochores (DNA segments with a fairly homogeneous base composition) for the three different tissues and the effects of isochores' base composition on their expression activity. Our analyses also cover the relations between the genes' expression activity and their localization in the isochore families. Conclusions: This study is the first where next-generation sequencing data are used to associate the effects of both genomic and genic compositional properties to their corresponding expression activity. Our findings confirm previous results, and further support the existence of a relationship between isochores and gene expression. This relationship corroborates that isochores are primarily a product of evolutionary adaptation rather than a simple by-product of neutral evolutionary processes. KW - Biased gene conversion KW - Human genome KW - GC-Content KW - Mammalian genomes KW - Base composition KW - Expresses genes KW - Higher rates KW - RNA-SEQ KW - Evolution KW - Rodents Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142773 VL - 12 IS - 511 ER - TY - JOUR A1 - Rauert, H. A1 - Stühmer, T. A1 - Bargou, R. A1 - Wajant, H. A1 - Siegmund, D. T1 - TNFR1 and TNFR2 regulate the extrinsic apoptotic pathway in myeloma cells by multiple mechanisms JF - Cell Death and Disease N2 - The huge majority of myeloma cell lines express TNFR2 while a substantial subset of them failed to show TNFR1 expression. Stimulation of TNFR1 in the TNFR1-expressing subset of MM cell lines had no or only a very mild effect on cellular viability. Surprisingly, however, TNF stimulation enhanced cell death induction by CD95L and attenuated the apoptotic effect of TRAIL. The contrasting regulation of TRAIL- and CD95L-induced cell death by TNF could be traced back to the concomitant NFjBmediated upregulation of CD95 and the antiapoptotic FLIP protein. It appeared that CD95 induction, due to its strength, overcompensated a rather moderate upregulation of FLIP so that the net effect of TNF-induced NFjB activation in the context of CD95 signaling is pro-apoptotic. TRAIL-induced cell death, however, was antagonized in response to TNF because in this context only the induction of FLIP is relevant. Stimulation of TNFR2 in myeloma cells leads to TRAF2 depletion. In line with this, we observed cell death induction in TNFR1-TNFR2-costimulated JJN3 cells. Our studies revealed that the TNF-TNF receptor system adjusts the responsiveness of the extrinsic apoptotic pathway in myeloma cells by multiple mechanisms that generate a highly context-dependent net effect on myeloma cell survival KW - apoptosis KW - CD95 KW - multiple myeloma KW - NFkB KW - TNF KW - TRAIL KW - NF-Kappa-B KW - Tumor-necrosis-factor KW - Factor receptor KW - Factor-alpha KW - Activation KW - Polymorphisms KW - Inhibitor KW - Promoter KW - Transcription KW - Expression Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-133486 VL - 2 ER - TY - JOUR A1 - Schwede, Angela A1 - Jones, Nicola A1 - Engstler, Markus A1 - Carrington, Mark T1 - The VSG C-terminal domain is inaccessible to antibodies on live trypanosomes JF - Molecular & Biochemical Parasitology N2 - In the mammalian host, the Trypanosoma brucei cell surface is covered with a densely packed protein coat of a single protein, the variant surface glycoprotein (VSG). The VSG is believed to shield invariant surface proteins from host antibodies but there is limited information on how far antibodies can penetrate into the VSG monolayer. Here, the VSG surface coat was probed to determine whether it acts as a barrier to binding of antibodies to the membrane proximal VSG C-terminal domain. The binding of C-terminal domain antibodies to VSG221 or VSG118 was compared with antibodies recognising the cognate whole VSGs. The C-terminal VSG domain was inaccessible to antibodies on live cells but not on fixed cells. This provides further evidence that the VSG coat acts as a barrier and protects the cell from antibodies that would otherwise bind to some of the other externally disposed proteins. KW - Trypanosome KW - VSG KW - Trypanosoma brucei KW - Cell surface KW - Antibody Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142746 VL - 175 IS - 2 ER - TY - JOUR A1 - Thormann, Birthe A1 - Raupach, Michael J. A1 - Wagner, Thomas A1 - Wägele, Johann W. A1 - Peters, Marcell K. T1 - Testing a Short Nuclear Marker for Inferring Staphylinid Beetle Diversity in an African Tropical Rain Forest JF - PLoS ONE N2 - Background: The use of DNA based methods for assessing biodiversity has become increasingly common during the last years. Especially in speciose biomes as tropical rain forests and/or in hyperdiverse or understudied taxa they may efficiently complement morphological approaches. The most successful molecular approach in this field is DNA barcoding based on cytochrome c oxidase I (COI) marker, but other markers are used as well. Whereas most studies aim at identifying or describing species, there are only few attempts to use DNA markers for inventorying all animal species found in environmental samples to describe variations of biodiversity patterns. Methodology/Principal Findings: In this study, an analysis of the nuclear D3 region of the 28S rRNA gene to delimit species-like units is compared to results based on distinction of morphospecies. Data derived from both approaches are used to assess diversity and composition of staphylinid beetle communities of a Guineo-Congolian rain forest in Kenya. Beetles were collected with a standardized sampling design across six transects in primary and secondary forests using pitfall traps. Sequences could be obtained of 99% of all individuals. In total, 76 molecular operational taxonomic units (MOTUs) were found in contrast to 70 discernible morphospecies. Despite this difference both approaches revealed highly similar biodiversity patterns, with species richness being equal in primary and secondary forests, but with divergent species communities in different habitats. The D3-MOTU approach proved to be an efficient tool for biodiversity analyses. Conclusions/Significance: Our data illustrate that the use of MOTUs as a proxy for species can provide an alternative to morphospecies identification for the analysis of changes in community structure of hyperdiverse insect taxa. The efficient amplification of the D3-marker and the ability of the D3-MOTUs to reveal similar biodiversity patterns as analyses of morphospecies recommend its use in future molecular studies on biodiversity. KW - DNA barcodes KW - Biological identifications KW - Species richness KW - Taxonomy KW - Conservation KW - Coleoptera KW - Parataxonomy KW - Assemblages KW - Madagascar Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142666 VL - 6 IS - 3 ER - TY - JOUR A1 - Waldholm, Johan A1 - Wang, Zhi A1 - Brodin, David A1 - Tyagi, Anu A1 - Yu, Simei A1 - Theopold, Ulrich A1 - Östlund Farrants, Ann Kristin A1 - Visa, Neus T1 - SWI/SNF regulates the alternative processing of a specific subset of pre-mRNAs in \(Drosophila\) \(melanogaster\) JF - BMC Molecular Biology N2 - Background: The SWI/SNF chromatin remodeling factors have the ability to remodel nucleosomes and play essential roles in key developmental processes. SWI/SNF complexes contain one subunit with ATPase activity, which in Drosophila melanogaster is called Brahma (Brm). The regulatory activities of SWI/SNF have been attributed to its influence on chromatin structure and transcription regulation, but recent observations have revealed that the levels of Brm affect the relative abundances of transcripts that are formed by alternative splicing and/or polyadenylation of the same pre-mRNA. Results: We have investigated whether the function of Brm in pre-mRNA processing in Drosophila melanogaster is mediated by Brm alone or by the SWI/SNF complex. We have analyzed the effects of depleting individual SWI/SNF subunits on pre-mRNA processing throughout the genome, and we have identified a subset of transcripts that are affected by depletion of the SWI/SNF core subunits Brm, Snr1 or Mor. The fact that depletion of different subunits targets a subset of common transcripts suggests that the SWI/SNF complex is responsible for the effects observed on pre-mRNA processing when knocking down Brm. We have also depleted Brm in larvae and we have shown that the levels of SWI/SNF affect the pre-mRNA processing outcome in vivo. Conclusions: We have shown that SWI/SNF can modulate alternative pre-mRNA processing, not only in cultured cells but also in vivo. The effect is restricted to and specific for a subset of transcripts. Our results provide novel insights into the mechanisms by which SWI/SNF regulates transcript diversity and proteomic diversity in higher eukaryotes. KW - Chromatin-remodeling complexes KW - In-vivo KW - Genes KW - Distinct KW - Brahma KW - Transcription KW - Trithorax KW - Subunit KW - Exons KW - BRM Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142613 VL - 12 IS - 46 ER - TY - JOUR A1 - Pham, Mirko A1 - Helluy, Xavier A1 - Kleinschnitz, Christoph A1 - Kraft, Peter A1 - Bartsch, Andreas J. A1 - Jakob, Peter A1 - Nieswandt, Bernhard A1 - Bendszus, Martin A1 - Guido, Stoll T1 - Sustained Reperfusion after Blockade of Glycoprotein-Receptor-Ib in Focal Cerebral Ischemia: An MRI Study at 17.6 Tesla JF - PLoS ONE N2 - Background: Inhibition of early platelet adhesion by blockade of glycoprotein-IB (GPIb) protects mice from ischemic stroke. To elucidate underlying mechanisms in-vivo, infarct development was followed by ultra-high field MRI at 17.6 Tesla. Methods: Cerebral infarction was induced by transient-middle-cerebral-artery-occlusion (tMCAO) for 1 hour in C57/BL6 control mice (N = 10) and mice treated with 100 mg Fab-fragments of the GPIb blocking antibody p0p/B 1 h after tMCAO (N = 10). To control for the effect of reperfusion, additional mice underwent permanent occlusion and received anti-GPIb treatment (N = 6; pMCAO) or remained without treatment (N = 3; pMCAO). MRI 2 h and 24 h after MCAO measured cerebral-blood-flow (CBF) by continuous arterial-spin labelling, the apparent-diffusion-coefficient (ADC), quantitative-T2 and T2-weighted imaging. All images were registered to a standard mouse brain MRI atlas and statistically analysed voxel-wise, and by cortico-subcortical ROI analysis. Results: Anti-GPIb treatment led to a relative increase of postischemic CBF vs. controls in the cortical territory of the MCA (2 h: 44.2 +/- 6.9 ml/100g/min versus 24 h: 60.5 +/- 8.4; p = 0.0012, F((1,18)) = 14.63) after tMCAO. Subcortical CBF 2 h after tMCAO was higher in anti-GPIb treated animals (45.3 +/- 5.9 vs. controls: 33.6 +/- 4.3; p = 0.04). In both regions, CBF findings were clearly related to a lower probability of infarction (Cortex/Subcortex of treated group: 35%/65% vs. controls: 95%/100%) and improved quantitative-T2 and ADC. After pMCAO, anti-GPIb treated mice developed similar infarcts preceded by severe irreversible hypoperfusion as controls after tMCAO indicating dependency of stroke protection on reperfusion. Conclusion: Blockade of platelet adhesion by anti-GPIb-Fab-fragments results in substantially improved CBF early during reperfusion. This finding was in exact spatial correspondence with the prevention of cerebral infarction and indicates in-vivo an increased patency of the microcirculation. Thus, progression of infarction during early ischemia and reperfusion can be mitigated by anti-platelet treatment. KW - Von-Willebrand-factor KW - Experimental stroke KW - Magnetic-resonance KW - Arterial water KW - Brain KW - Perfusion KW - Mice KW - Inflammation KW - Coefficient KW - mechanisms Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142608 VL - 6 IS - 4 ER - TY - JOUR A1 - Müller-Sienerth, Nicole A1 - Dietz, Lena A1 - Holtz, Philipp A1 - Kapp, Markus A1 - Grigoleit, Götz Ulrich A1 - Schmuck, Carsten A1 - Wajant, Harald A1 - Siegmund, Daniela T1 - SMAC Mimetic BV6 Induces Cell Death in Monocytes and Maturation of Monocyte-Derived Dendritic Cells JF - PLoS ONE N2 - Background: Compounds mimicking the inhibitory effect of SMAC / DIABLO on X-linked inhibitor of apoptosis (XIAP) have been developed with the aim to achieve sensitization for apoptosis of tumor cells resistant due to deregulated XIAP expression. It turned out that SMAC mimetics also have complex effects on the NF kappa B system and TNF signaling. In view of the overwhelming importance of the NF kappa B transcription factors in the immune system, we analyzed here the effects of the SMAC mimetic BV6 on immune cells. Principal Findings: BV6 induced apoptotic and necrotic cell death in monocytes while T-cells, dendritic cells and macrophages were largely protected against BV6-induced cell death. In immature dendritic cells BV6 treatment resulted in moderate activation of the classical NF kappa B pathway, but it also diminished the stronger NF kappa B-inducing effect of TNF and CD40L. Despite its inhibitory effect on TNF- and CD40L signaling, BV6 was able to trigger maturation of immature DCs as indicated by upregulation of CD83, CD86 and IL12. Significance: The demonstrated effects of SMAC mimetics on immune cells may complicate the development of tumor therapeutic concepts based on these compounds but also arise the possibility to exploit them for the development of immune stimulatory therapies. KW - Kappa-B activation KW - Alpha-dependent apoptosis KW - Caspase-8 activation KW - Cancer KW - TRAF2 KW - CIAP1 KW - Necrosis KW - Complex KW - C-IAP1 KW - RIP1 Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142415 VL - 6 IS - 6 ER - TY - JOUR A1 - Bergmiller, Tobias A1 - Pena-Miller, Rafael A1 - Boehm, Alexander A1 - Ackermann, Martin T1 - Single-cell time-lapse analysis of depletion of the universally conserved essential protein YgjD JF - BMC Microbiology N2 - Background: The essential Escherichia coli gene ygjD belongs to a universally conserved group of genes whose function has been the focus of a number of recent studies. Here, we put ygjD under control of an inducible promoter, and used time-lapse microscopy and single cell analysis to investigate the phenotypic consequences of the depletion of YgjD protein from growing cells. Results: We show that loss of YgjD leads to a marked decrease in cell size and termination of cell division. The transition towards smaller size occurs in a controlled manner: cell elongation and cell division remain coupled, but cell size at division decreases. We also find evidence that depletion of YgjD leads to the synthesis of the intracellular signaling molecule (p) ppGpp, inducing a cellular reaction resembling the stringent response. Concomitant deletion of the relA and spoT genes - leading to a strain that is uncapable of synthesizing (p) ppGpp abrogates the decrease in cell size, but does not prevent termination of cell division upon YgjD depletion. Conclusions: Depletion of YgjD protein from growing cells leads to a decrease in cell size that is contingent on (p) ppGpp, and to a termination of cell division. The combination of single-cell time-lapse microscopy and statistical analysis can give detailed insights into the phenotypic consequences of the loss of essential genes, and can thus serve as a new tool to study the function of essential genes. KW - Transfer-RNA modification KW - Escherichia-coli K-12 KW - Gene KW - Division KW - Expression KW - Inactivation KW - Maintenance KW - Growth KW - Level KW - Ftsz Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142324 VL - 11 IS - 118 ER - TY - JOUR A1 - Haeussinger, Florian B. A1 - Heinzel, Sebastian A1 - Hahn, Tim A1 - Schecklmann, Martin A1 - Ehlis, Ann-Christine A1 - Fallgatter, Andreas J. T1 - Simulation of Near-Infrared Light Absorption Considering Individual Head and Prefrontal Cortex Anatomy: Implications for Optical Neuroimaging JF - PLoS ONE N2 - Functional near-infrared spectroscopy (fNIRS) is an established optical neuroimaging method for measuring functional hemodynamic responses to infer neural activation. However, the impact of individual anatomy on the sensitivity of fNIRS measuring hemodynamics within cortical gray matter is still unknown. By means of Monte Carlo simulations and structural MRI of 23 healthy subjects (mean age: (25.0 +/- 2.8) years), we characterized the individual distribution of tissue-specific NIR-light absorption underneath 24 prefrontal fNIRS channels. We, thereby, investigated the impact of scalp-cortex distance (SCD), frontal sinus volume as well as sulcal morphology on gray matter volumes (V(gray)) traversed by NIR-light, i.e. anatomy-dependent fNIRS sensitivity. The NIR-light absorption between optodes was distributed describing a rotational ellipsoid with a mean penetration depth of (23.6 +/- 0.7) mm considering the deepest 5% of light. Of the detected photon packages scalp and bone absorbed (96.4 +/- 9: 7)% and V(gray) absorbed (3.1 +/- 1.8)% of the energy. The mean V(gray) volume (1.1 +/- 0.4)cm(3) was negatively correlated (r = - .76) with the SCD and frontal sinus volume (r = - .57) and was reduced by 41.5% in subjects with relatively large compared to small frontal sinus. Head circumference was significantly positively correlated with the mean SCD (r = .46) and the traversed frontal sinus volume (r = .43). Sulcal morphology had no significant impact on V(gray). Our findings suggest to consider individual SCD and frontal sinus volume as anatomical factors impacting fNIRS sensitivity. Head circumference may represent a practical measure to partly control for these sources of error variance. KW - Beer-lambert law KW - Adult head KW - Human brain KW - Spectroscopy fnirs KW - Photon migration KW - Propagation KW - Scattering KW - Model KW - Tissues KW - Media Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142311 VL - 6 IS - 10 ER - TY - JOUR A1 - Chen, Nanhai G. A1 - Yu, Yong A. A1 - Zhang, Qian A1 - Szalay, Aladar A. T1 - Replication efficiency of oncolytic vaccinia virus in cell cultures prognosticates the virulence and antitumor efficacy in mice JF - Journal of Translational Medicine N2 - Background: We have shown that insertion of the three vaccinia virus (VACV) promoter-driven foreign gene expression cassettes encoding Renilla luciferase-Aequorea GFP fusion protein, beta-galactosidase, and beta-glucuronidase into the F14.5L, J2R, and A56R loci of the VACV LIVP genome, respectively, results in a highly attenuated mutant strain GLV 1h68. This strain shows tumor specific replication and is capable of eradicating tumors with little or no virulence in mice. This study aimed to distinguish the contribution of added VACV promoter-driven transcriptional units as inserts from the effects of insertional inactivation of three viral genes, and to determine the correlation between replication efficiency of oncolytic vaccinia virus in cell cultures and the virulence and antitumor efficacy in mice Methods: A series of recombinant VACV strains was generated by replacing one, two, or all three of the expression cassettes in GLV 1h68 with short non coding DNA sequences. The replication efficiency and tumor cell killing capacity of these newly generated VACV strains were compared with those of the parent virus GLV-1h68 in cell cultures. The virus replication efficiency in tumors and antitumor efficacy as well as the virulence were evaluated in nu/nu (nude) mice bearing human breast tumor xenografts. Results: we found that virus replication efficiency increased with removal of each of the expression cassettes. The increase in virus replication efficiency was proportionate to the strength of removed VACV promoters linked to foreign genes. The replication efficiency of the new VACV strains paralleled their cytotoxicity in cell cultures. The increased replication efficiency in tumor xenografts resulted in enhanced antitumor efficacy in nude mice. Similarly, the enhanced virus replication efficiency was indicative of increased virulence in nude mice. Conclusions: These data demonstrated that insertion of VACV promoter-driven transcriptional units into the viral genome for the purpose of insertional mutagenesis did modulate the efficiency of virus replication together with antitumor efficacy as well as virulence. Replication efficiency of oncolytic VACV in cell cultures can predict the virulence and therapeutic efficacy in nude mice. These findings may be essential for rational design of safe and potent VACV strains for vaccination and virotherapy of cancer in humans and animals. KW - Recombinant vaccinia KW - Nude-mice KW - Cancer KW - GLV-1H68 KW - Therapy KW - Agent KW - Regression KW - Carcinoma KW - Deletion KW - Protein KW - modulation of virus replication KW - GI-101A tumor xenografts KW - oncolytic virotherapy Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142268 VL - 9 IS - 164 ER - TY - JOUR A1 - Hartl, Maximilian J. A1 - Bodem, Jochen A1 - Jochheim, Fabian A1 - Rethwilm, Axel A1 - Rösch, Paul A1 - Wöhrl, Birgitta M. T1 - Regulation of foamy virus protease activity by viral RNA JF - Retrovirology N2 - No abstract available. KW - Virologie Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142248 VL - 8 IS - Suppl. 1 ER - TY - JOUR A1 - Weis, Eva A1 - Schoen, Holger A1 - Victor, Anja A1 - Spix, Claudia A1 - Ludwig, Marco A1 - Schneider-Raetzke, Brigitte A1 - Kohlschmidt, Nicolai A1 - Bartsch, Oliver A1 - Gerhold-Ay, Aslihan A1 - Boehm, Nils A1 - Grus, Franz A1 - Haaf, Thomas A1 - Galetzka, Danuta T1 - Reduced mRNA and Protein Expression of the Genomic Caretaker RAD9A in Primary Fibroblasts of Individuals with Childhood and Independent Second Cancer JF - PLoS ONE N2 - Background: The etiology of secondary cancer in childhood cancer survivors is largely unclear. Exposure of normal somatic cells to radiation and/or chemotherapy can damage DNA and if not all DNA lesions are properly fixed, the mis-repair may lead to pathological consequences. It is plausible to assume that genetic differences, i.e. in the pathways responsible for cell cycle control and DNA repair, play a critical role in the development of secondary cancer. Methodology/Findings: To identify factors that may influence the susceptibility for second cancer formation, we recruited 20 individuals who survived a childhood malignancy and then developed a second cancer as well as 20 carefully matched control individuals with childhood malignancy but without a second cancer. By antibody microarrays, we screened primary fibroblasts of matched patients for differences in the amount of representative DNA repair-associated proteins. We found constitutively decreased levels of RAD9A and several other DNA repair proteins in two-cancer patients, compared to one-cancer patients. The RAD9A protein level increased in response to DNA damage, however to a lesser extent in the two-cancer patients. Quantification of mRNA expression by real-time RT PCR revealed lower RAD9A mRNA levels in both untreated and 1 Gy gamma-irradiated cells of two-cancer patients. Conclusions/Significance: Collectively, our results support the idea that modulation of RAD9A and other cell cycle arrest and DNA repair proteins contribute to the risk of developing a second malignancy in childhood cancer patients. KW - DNA methylation KW - Malignant neoplasms KW - Genes KW - Instability KW - Stability KW - Susceptibility KW - Checkpoints KW - Repair KW - Damage Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141838 VL - 6 IS - 10 ER - TY - JOUR A1 - Werner, Katharina A1 - Schwede, Frank A1 - Genieser, Hans-Gottfried A1 - Geiger, Jörg A1 - Butt, Elke T1 - Quantification of cAMP and cGMP analogs in intact cells: pitfalls in enzyme immunoassays for cyclic nucleotides JF - Naunyn-Schmiedeberg's Archives of Pharmacology N2 - Immunoassays are routinely used as research tools to measure intracellular cAMP and cGMP concentrations. Ideally, this application requires antibodies with high sensitivity and specificity. The present work evaluates the cross-reactivity of commercially available cyclic nucleotide analogs with two non-radioactive and one radioactive cAMP and cGMP immunoassay. Most of the tested cyclic nucleotide analogs showed low degree competition with the antibodies; however, with Rp-cAMPS, 8-Br-cGMP and 8-pCPT-cGMP, a strong cross-reactivity with the corresponding cAMP and cGMP, respectively, immunoassays was observed. The determined EIA-binding constants enabled the measurement of the intracellular cyclic nucleotide concentrations and revealed a time- and lipophilicity-dependent cell membrane permeability of the compounds in the range of 10–30% of the extracellular applied concentration, thus allowing a more accurate prediction of the intracellular analog levels in a given experiment. KW - Cyclic nucleotides KW - Enzyme immunoassay KW - Lipophilicity KW - Cell permeability Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141828 VL - 384 IS - 2 ER - TY - JOUR A1 - Schöttker, Björn A1 - Schmidt-Wolf, Ingo G. H. T1 - Pulsing with blast cell lysate or blast-derived total RNA reverses the dendritic cell-mediated cytotoxic activity of cytokine-induced killer cells against allogeneic acute myelogenous leukemia cells T1 - Pulsen mit Blastenzelllysat oder Blasten-Gesamt-RNA richtet die durch dendritische Zellen vermittelte Aktivität von Zytokin-induzierten Killerzellen gegen allogene akute myeloische Zellen JF - GMS German Medical Science N2 - Immunotherapeutic strategies may be a treatment option in patients with refractory acute myelogenous leukemia (AML) or, in cases of complete remission after conventional therapy regimens, may help to reduce disease recurrence or delay time to progression. Evidence suggests a key role of dendritic cells (DCs) in cancer immunotherapy due to their capacity to present tumour antigens to effector cells. We generated cytokine-induced killer (CIK) cells from healthy donors and examined their responses in vitro in an LDH release assay against three cell lines and allogeneic HLA non-matched blasts from three patients with de novo AML after coincubation with autologous peripheral blood monocyte-derived DCs. Although DCs were unable to enhance CIK cell effects against all three cell lines tested, the cytotoxic activity against the patients’ AML cells increased after coculture with mature DCs, which was significant in two of three patients. However, neither prior pulsing of the DCs with blast cell lysates nor with leukemic cell-derived total RNA further enhanced the lytic capacity of the CIK cells. On the contrary, pulsing reduced or even reversed the cytotoxic activity of the effector cells. This decrease of allogeneic cytotoxicity led us to conclude that monocyte-derived DCs may be useful in autologous or allogeneic vaccine strategies for the treatment of AML or in priming donor lymphocytes in vitro, but unfractionated antigens as pulsing agents may have inhibitory effects on T cell efficiency and their employment in immunotherapeutic strategies for AML seems questionable. N2 - Immuntherapeutische Strategien können eine Behandlungsoption bei Patienten mit refraktärer akuter myeloischer Leukämie (AML) sein oder in den Fällen einer kompletten Remission nach konventionellen Therapieformen helfen, das Wiederauftreten der Krankheit zu verhindern oder die Zeit bis zur Progression zu verlängern. Es gibt Hinweise darauf, dass dendritische Zellen (DCs) eine zentrale Rolle in der Krebs-Immuntherapie spielen aufgrund ihrer Fähigkeit, tumorantigene Effektor-Zellen zu präsentieren. Wir stellten Zytokin-induzierte Killer (CIK)-Zellen von gesunden Spendern her und untersuchten deren Reaktionen in vitro in einem Laktatdehydrogenase (LDH)-Assay gegen Zelllinien und allogene HLA nicht übereinstimmende Blasten von drei Patienten mit de novo AML nach Koinkubation mit autologen aus dem peripheren Blut abgeleiteten DCs. Obwohl DCs die CIK Zellen Wirksamkeit gegen alle drei getesteten Zelllinien nicht verbessern konnten, wurde die zytotoxische Aktivität gegen die Patienten-AML-Zellen nach Kokultur mit reifen DCs in zwei von drei Patienten signifikant erhöht. Doch weder ein Pulsen der DCs mit blastären Zelllysaten noch mit aus leukämischen Zellen gewonnener Gesamt-RNA konnten die lytische Kapazität der CIK-Zellen weiter verbessern. Im Gegenteil, gepulste DCs reduzierten sogar die zytotoxische Aktivität der Effektorzellen. Dieser Rückgang der allogenen Zytotoxizität führte uns zu dem Schluss, dass von Monozyten abgeleitete DCs nützlich sein könnten in autologen oder allogenen Impfstrategien zur Behandlung von AML. Unfraktionierte Antigene zum Pulsen von DC können dagegen hemmende Wirkung auf T-Zellen haben. KW - dendritic cells KW - AML KW - blast-derived RNA KW - Zytokin-induzierte Killerzellen KW - Zelllysat KW - cytokine-induced killer cells KW - blast cell lysate KW - dendritische Zellen KW - RNA Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:0183-0001410 VL - 9 IS - Doc18 ER - TY - JOUR A1 - Goepel, Johanna A1 - Biehl, Stefanie C. A1 - Kissler, Johanna A1 - Paul-Jordanov, Isabelle T1 - Pro- and antisaccades in children elicited by visual and acoustic targets - does modality matter? JF - BMC Pediatrics N2 - Background: Children are able to inhibit a prepotent reaction to suddenly arising visual stimuli, although this skill is not yet as pronounced as it is in adulthood. However, up to now the inhibition mechanism to acoustic stimuli has been scarcely investigated Methods: Reflexive (prosaccade) and inhibitory (antisaccade) responses to visual and acoustic targets were examined with an eye tracker system in 31 children between seven and twelve years of age using a gap-overlap task and two target eccentricities. Results: Acoustically cued saccades had longer reaction times than visually cued saccades. A gap effect (i.e., shorter reaction time in the gap than the overlap condition) was only found for visually elicited saccades, whereas an eccentricity effect (i.e., faster saccades to more laterally presented targets - 12 degrees vs. 6 degrees or rather 90 degrees vs. 45 degrees) was only present in the acoustic condition. Longer reaction times of antisaccades compared to prosaccades were found only in the visual task. Across both tasks the typical pattern of elevated error rates in the antisaccade condition was found. Antisaccade errors declined with age, indicating an ongoing development of inhibitory functions. Conclusions: The present results lay the ground for further studies of acoustically triggered saccades in typically as well as atypically developing children and it might thus be possible to upgrade physiological diagnostic tools. KW - Saccadic eye-movements KW - Auditory targets KW - Voluntary control KW - Task-performance KW - Latency KW - Prosaccade KW - Vergence KW - Stimuli KW - GAP Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141807 VL - 11 IS - 116 ER - TY - JOUR A1 - Nell, Manuel A1 - Burgkart, Rainer H. A1 - Gradl, Guntmar A1 - von Eisenhart-Rothe, Rüdiger A1 - Schaeffeler, Christoph A1 - Trappe, Dennis A1 - Prazeres da Costa, Clarissa A1 - Gradinger, Reiner A1 - Kirchhoff, Chlodwig T1 - Primary extrahepatic alveolar echinococcosis of the lumbar spine and the psoas muscle JF - Annals of Clinical Microbiology and Antimicrobials N2 - Alveolar echinococcosis (AE) of human being caused by Echinococcus multilocularis is a rare but important zoonosis especially in tempered zones of middle Europe and Northern America with endemic character in many countries. Due to the long incubation period, various clinical manifestations, critical prognosis, and outcome AE presents a serious and severe disease. The primary focus of infection is usually the liver. Although secondary affection of visceral organs is possible extrahepatic AE is highly uncommon. Moreover, the involvement of bone and muscle presents with an even lower incidence. In the literature numerous cases on hepatic AE have been reported. However, extrahepatic AE involving bones and/or muscles was described very rarely. We report a case of an 80-year-old man with primary extrahepatic alveolar Echinococcosis of the lumbar spine and the psoas muscle. The etiology, diagnosis, differential diagnoses, treatment options and outcome of this rare disease are discussed in context with the current literature. KW - Medicine Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141796 VL - 10 IS - 13 ER - TY - JOUR A1 - Stelzig, Yvonne A1 - Jacob, Roland A1 - Mueller, Joachim T1 - Preliminary speech recognition results after cochlear implantation in patients with unilateral hearing loss: a case series JF - Journal of Medical Case Reports N2 - Introduction Cochlear implants known to provide support in individuals with bilateral hearing loss may also be of great benefit for individuals with unilateral hearing loss. This case report demonstrates the positive effects of cochlear implantation on speech understanding in noise conditions in patients with unilateral hearing loss and normal hearing on the contralateral side. To the best of our knowledge, the data presented here are from the first few cases to receive a cochlear implant for unilateral hearing loss. Case presentation Four Caucasian German men, two aged 48 and the others aged 51 and 57 years old, with post-lingual unilateral hearing loss and normal hearing on the contralateral side were implanted with a cochlear implant. All our patients were members of the German army. Before and after implantation, they were given a battery of speech tests in different hearing conditions to assess the effect of unilateral cochlear implantation on speech understanding in noise conditions. Test results showed that all patients benefited from unilateral cochlear implantation, particularly in terms of speech understanding in noise conditions. Conclusions Unilateral cochlear implantation might be a successful treatment method for patients with unilateral hearing loss not benefiting from alternative treatment options. The results of this case report open up the field of cochlear implantation for expanded criteria and new areas of research. KW - medicin Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141722 VL - 5 IS - 343 ER -