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Chronic myeloid leukaemia (CML) is a clonal myeloproliferative stem cell disorder characterized by the constitutively active BCR‐ABL tyrosine kinase. The LIM and SH3 domain protein 1 (LASP1) has recently been identified as a novel BCR‐ABL substrate and is associated with proliferation, migration, tumorigenesis and chemoresistance in several cancers. Furthermore, LASP1 was shown to bind to the chemokine receptor 4 (CXCR4), thought to be involved in mechanisms of relapse. In order to identify potential LASP1‐mediated pathways and related factors that may help to further eradicate minimal residual disease (MRD), the effect of LASP1 on processes involved in progression and maintenance of CML was investigated. The present data indicate that not only overexpression of CXCR4, but also knockout of LASP1 contributes to proliferation, reduced apoptosis and migration as well as increased adhesive potential of K562 CML cells. Furthermore, LASP1 depletion in K562 CML cells leads to decreased cytokine release and reduced NK cell‐mediated cytotoxicity towards CML cells. Taken together, these results indicate that in CML, reduced levels of LASP1 alone and in combination with high CXCR4 expression may contribute to TKI resistance.
Background
Neisseria meningitidis is a naturally transformable, facultative pathogen colonizing the human nasopharynx. Here, we analyze on a genome-wide level the impact of recombination on gene-complement diversity and virulence evolution in N. meningitidis. We combined comparative genome hybridization using microarrays (mCGH) and multilocus sequence typing (MLST) of 29 meningococcal isolates with computational comparison of a subset of seven meningococcal genome sequences.
Principal Findings
We found that lateral gene transfer of minimal mobile elements as well as prophages are major forces shaping meningococcal population structure. Extensive gene content comparison revealed novel associations of virulence with genetic elements besides the recently discovered meningococcal disease associated (MDA) island. In particular, we identified an association of virulence with a recently described canonical genomic island termed IHT-E and a differential distribution of genes encoding RTX toxin- and two-partner secretion systems among hyperinvasive and non-hyperinvasive lineages. By computationally screening also the core genome for signs of recombination, we provided evidence that about 40% of the meningococcal core genes are affected by recombination primarily within metabolic genes as well as genes involved in DNA replication and repair. By comparison with the results of previous mCGH studies, our data indicated that genetic structuring as revealed by mCGH is stable over time and highly similar for isolates from different geographic origins.
Conclusions
Recombination comprising lateral transfer of entire genes as well as homologous intragenic recombination has a profound impact on meningococcal population structure and genome composition. Our data support the hypothesis that meningococcal virulence is polygenic in nature and that differences in metabolism might contribute to virulence.
Virotherapy using oncolytic vaccinia virus strains is one of the most promising new strategies for cancer therapy. In this study, we analyzed for the first time the therapeutic efficacy of the oncolytic vaccinia virus GLV-1h68 in two human hepatocellular carcinoma cell lines HuH7 and PLC/PRF/5 (PLC) in cell culture and in tumor xenograft models. By viral proliferation assays and cell survival tests, we demonstrated that GLV-1h68 efficiently colonized, replicated in, and did lyse these cancer cells in culture. Experiments with HuH7 and PLC xenografts have revealed that a single intravenous injection (i.v.) of mice with GLV-1h68 resulted in a significant reduction of primary tumor sizes compared to uninjected controls. In addition, replication of GLV-1h68 in tumor cells led to strong inflammatory and oncolytic effects resulting in intense infiltration of MHC class II-positive cells like neutrophils, macrophages, B cells and dendritic cells and in up-regulation of 13 pro-inflammatory cytokines. Furthermore, GLV-1h68 infection of PLC tumors inhibited the formation of hemorrhagic structures which occur naturally in PLC tumors. Interestingly, we found a strongly reduced vascular density in infected PLC tumors only, but not in the non-hemorrhagic HuH7 tumor model. These data demonstrate that the GLV-1h68 vaccinia virus may have an enormous potential for treatment of human hepatocellular carcinoma in man.
Das Ziel der vorliegenden Arbeit war es, einen Beitrag zu leisten zur Klärung der Beziehung zwischen Alexithymie und den subjektiven und physiologischen Reaktionen auf emotionale Situationen. Kern des Persönlichkeitsmerkmals ‘Alexithymie’ ist die Schwierigkeit, eigene Gefühle zu identifizieren und anderen mitzuteilen (Bagby & Taylor, 1999a). Ähnlich wie bei anderen Formen emotionaler Hemmung wurde auch bei Alexithymie eine erhöhte physiologische Reaktivität angenommen, die auch mit einem erhöhten Erkrankungsrisiko verbunden sein soll (Stress-Alexithymie Hypothese, Martin & Pihl, 1985). Demnach führt eine in Stresssituationen durch mangelnde Emotionsregulation erhöhte und verlängerte physiologische Aktivität bei alexithymen Personen zu körperlichen Erkrankungen. In der Entkopplungshypothese (Papciak, Feuerstein & Spiegel, 1985) geht man bei Alexithymie unspezifischer als bei der Stress-Alexithymie Hypothese von einer Dissoziation der physiologischen Aktivität und den subjektiven Angaben zu Gefühlen oder emotionaler Erregung aus. Zu diesen Hypothesen liegen jedoch nur wenige und zudem widersprüchliche empirische Befunde vor. Die zentrale Frage der vorliegenden Arbeit lautete daher, ob sich hoch und niedrig alexithyme Personen in ihren subjektiven und physiologischen Reaktionen auf emotionale und belastende Situationen unterscheiden. Dazu wurde je eine experimentelle Untersuchung mit gesunden Probanden (n=43) und mit Patienten einer psychosomatischen Klinik (n=82) durchgeführt. Alle Probanden wurden nach der 20-Item Toronto-Alexithymieskala (Bagby, Parker & Taylor, 1994) in hoch und niedrig alexithyme Personen eingeteilt. Nach der Induktion von Emotionen und Belastungen (durch Filmausschnitte, Hyperventilation und einen modifizierten Stroop-Test) wurden die Reaktionen der Versuchspersonen hinsichtlich ihrer Gefühle, Körperempfindungen und physiologischen Parameter erfasst. Wie erwartet berichteten hoch alexithyme Gesunde und besonders Patienten im Vergleich zu niedrig Alexithymen stärkere negative Emotionen (v.a. Angst) und in einigen Bereichen stärkere körperliche Empfindungen im tonischen Niveau (vor der Emotionsinduktion). Jedoch ergaben sich entgegen den Erwartungen keine Gruppenunterschiede in den physiologischen Variablen. Durch Darbietung von Filmausschnitten wurden die Zielemotionen Traurigkeit und Ärger in ausreichender Stärke induziert. Während der Filme zeigten hoch Alexithyme stärkere Angst als niedrig Alexithyme. Signifikante Unterschiede zwischen hoch und niedrig alexithymen Personen in den Zielemotionen der Filmausschnitte oder anderen Emotionen fanden sich jedoch nicht. Allerdings beurteilten in beiden Untersuchungen weniger hoch als niedrig alexithyme Personen die Zielemotion Traurigkeit als stärkste Emotion während der traurigkeitsinduzierenden Filme. Hoch alexithyme Gesunde und stärker noch Patienten berichteten stärkere körperliche Empfindungen sowie größere Schwierigkeiten, ihre Gefühle während der Filmausschnitte in Worte zu fassen. Signifikante Unterschiede in der physiologischen Reaktivität auf die Filmausschnitte waren jedoch nicht nachweisbar. Vergleichbare Ergebnisse wie bei der Emotionsinduktion zeigten sich ebenfalls bei körperlicher und kognitiver Belastung. Die Befunde der vorliegenden Untersuchungen gelten damit für emotionale Situationen sowie auch für körperliche und kognitive Belastungen. Weder die Vorhersagen der Stress-Alexithymie Hypothese noch die der Entkopplungshypothese konnten in den vorliegenden Untersuchungen bestätigt werden. Ingesamt sprechen die Befunde daher dafür, dass eine mögliche höhere Vulnerabilität alexithymer Personen für körperliche Krankheiten nicht auf eine verstärkte physiologische Reaktivität auf spezifische emotionale Situationen zurückzuführen ist. Die Ergebnisse weisen allerdings auf eine in der Entkopplungshypothese nicht postulierte Dissoziation zwischen der objektiv messbaren und der wahrgenommenen physiologischen Reaktivität bei hoch alexithymen Patienten hin. Die stärkere Fokussierung auf körperliche Empfindungen lässt einen verstärkten Bericht körperlicher Symptome sowie ein verstärktes Krankheitsverhalten dieser Patienten erwarten.
Recently, we have described novel pyridyl indole esters and peptidomimetics as potent inhibitors of the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) main protease. Here, we analysed the impact of these compounds on viral replication. It has been shown that some antivirals against SARS-CoV-2 act in a cell line-specific way. Thus, the compounds were tested in Vero, Huh-7, and Calu-3 cells. We showed that the protease inhibitors at 30 µM suppress viral replication by up to 5 orders of magnitude in Huh-7 cells, while in Calu-3 cells, suppression by 2 orders of magnitude was achieved. Three pyridin-3-yl indole-carboxylates inhibited viral replication in all cell lines, indicating that they might repress viral replication in human tissue as well. Thus, we investigated three compounds in human precision-cut lung slices and observed donor-dependent antiviral activity in this patient-near system. Our results provide evidence that even direct-acting antivirals may act in a cell line-specific manner.
Animal models point towards a key role of brain-derived neurotrophic factor (BDNF), insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor (VEGF) in mediating exercise-induced structural and functional changes in the hippocampus. Recently, also platelet derived growth factor-C (PDGF-C) has been shown to promote blood vessel growth and neuronal survival. Moreover, reductions of these neurotrophic and angiogenic factors in old age have been related to hippocampal atrophy, decreased vascularization and cognitive decline. In a 3-month aerobic exercise study, forty healthy older humans (60 to 77 years) were pseudo-randomly assigned to either an aerobic exercise group (indoor treadmill, n = 21) or to a control group (indoor progressive-muscle relaxation/stretching, n = 19). As reported recently, we found evidence for fitness-related perfusion changes of the aged human hippocampus that were closely linked to changes in episodic memory function. Here, we test whether peripheral levels of BDNF, IGF-I, VEGF or PDGF-C are related to changes in hippocampal blood flow, volume and memory performance. Growth factor levels were not significantly affected by exercise, and their changes were not related to changes in fitness or perfusion. However, changes in IGF-I levels were positively correlated with hippocampal volume changes (derived by manual volumetry and voxel-based morphometry) and late verbal recall performance, a relationship that seemed to be independent of fitness, perfusion or their changes over time. These preliminary findings link IGF-I levels to hippocampal volume changes and putatively hippocampus-dependent memory changes that seem to occur over time independently of exercise. We discuss methodological shortcomings of our study and potential differences in the temporal dynamics of how IGF-1, VEGF and BDNF may be affected by exercise and to what extent these differences may have led to the negative findings reported here.
Background:
Commensal bacteria like Neisseria meningitidis sometimes cause serious disease. However, genomic comparison of hyperinvasive and apathogenic lineages did not reveal unambiguous hints towards indispensable virulence factors. Here, in a systems biological approach we compared gene expression of the invasive strain MC58 and the carriage strain α522 under different ex vivo conditions mimicking commensal and virulence compartments to assess the strain-specific impact of gene regulation on meningococcal virulence.
Results:
Despite indistinguishable ex vivo phenotypes, both strains differed in the expression of over 500 genes under infection mimicking conditions. These differences comprised in particular metabolic and information processing genes as well as genes known to be involved in host-damage such as the nitrite reductase and numerous LOS biosynthesis genes. A model based analysis of the transcriptomic differences in human blood suggested ensuing metabolic flux differences in energy, glutamine and cysteine metabolic pathways along with differences in the activation of the stringent response in both strains. In support of the computational findings, experimental analyses revealed differences in cysteine and glutamine auxotrophy in both strains as well as a strain and condition dependent essentiality of the (p)ppGpp synthetase gene relA and of a short non-coding AT-rich repeat element in its promoter region.
Conclusions:
Our data suggest that meningococcal virulence is linked to transcriptional buffering of cryptic genetic variation in metabolic genes including global stress responses. They further highlight the role of regulatory elements for bacterial virulence and the limitations of model strain approaches when studying such genetically diverse species as N. meningitidis.
Some members of the physiological human microbiome occasionally cause life-threatening disease even in immunocompetent individuals. A prime example of such a commensal pathogen is Neisseria meningitidis, which normally resides in the human nasopharynx but is also a leading cause of sepsis and epidemic meningitis. Using N. meningitidis as model organism, we tested the hypothesis that virulence of commensal pathogens is a consequence of within host evolution and selection of invasive variants due to mutations at contingency genes, a mechanism called phase variation. In line with the hypothesis that phase variation evolved as an adaptation to colonize diverse hosts, computational comparisons of all 27 to date completely sequenced and annotated meningococcal genomes retrieved from public databases showed that contingency genes are indeed enriched for genes involved in host interactions. To assess within-host genetic changes in meningococci, we further used ultra-deep whole-genome sequencing of throat-blood strain pairs isolated from four patients suffering from invasive meningococcal disease. We detected up to three mutations per strain pair, affecting predominantly contingency genes involved in type IV pilus biogenesis. However, there was not a single (set) of mutation(s) that could invariably be found in all four pairs of strains. Phenotypic assays further showed that these genetic changes were generally not associated with increased serum resistance, higher fitness in human blood ex vivo or differences in the interaction with human epithelial and endothelial cells in vitro. In conclusion, we hypothesize that virulence of meningococci results from accidental emergence of invasive variants during carriage and without within host evolution of invasive phenotypes during disease progression in vivo.