TY - THES A1 - Zude, Ingmar T1 - Characterization of virulence-associated traits of Escherichia coli bovine mastitis isolates T1 - Charakterisierung Virulenz-assozierter Eigenschaften von Escherichia coli Isolaten boviner Mastitis N2 - Bacterial mastitis is caused by invasion of the udder, bacterial multiplication and induction of inflammatory responses in the bovine mammary gland. Disease severity and the cause of disease are influenced by environmental factors, the cow’s immune response as well as bacterial traits. Escherichia coli (E. coli) is one of the main causes of acute bovine mastitis, but although pathogenic E. coli strains can be classified into different pathotypes, E. coli causing mastitis cannot unambiguously be distinguished from commensal E. coli nor has a common set of virulence factors been described for mastitis isolates. This project focussed on the characterization of virulence- associated traits of E. coli mastitis isolates in comprehensive analyses under conditions either mimicking initial pathogenesis or conditions that E. coli mastitis isolates should encounter while entering the udder. Virulence-associated traits as well as fitness traits of selected bovine mastitis or faecal E. coli strains were identified and analyzed in comparative phenotypic assays. Raw milk whey was introduced to test bacterial fitness in native mammary secretion known to confer antimicrobial effects. Accordingly, E. coli isolates from bovine faeces represented a heterogeneous group of which some isolates showed reduced ability to survive in milk whey whereas others phenotypically resembled mastitis isolates that represented a homogeneous group in that they showed similar survival and growth characteristics in milk whey. In contrast, mastitis isolates did not exhibit such a uniform phenotype when challenged with iron shortage, lactose as sole carbon source and lingual antimicrobial peptide (LAP) as a main defensin of milk. Reduced bacterial fitness could be related to LAP suggesting that bacterial adaptation to an intramammary lifestyle requires resistance to host defensins present in mammary secretions, at least LAP. E. coli strain 1303 and ECC-1470 lack particular virulence genes associated to mastitis isolates. To find out whether differences in gene expression may contribute to the ability of E. coli variants to cause mastitis, the transcriptome of E. coli model mastitis isolates 1303 and ECC-1470 were analyzed to identify candidate genes involved in bacterium-host interaction, fitness or even pathogenicity during bovine mastitis. DNA microarray analysis was employed to assess the transcriptional response of E. coli 1303 and ECC-1470 upon cocultivation with MAC-T immortalized bovine mammary gland epithelial cells to identify candidate genes involved in bacterium-host interaction. Additionally, the cell adhesion and invasion ability of E. coli strain 1303 and ECC-1470 was investigated. The transcriptonal response to the presence of host cells rather suggested competition for nutrients and oxygen between E. coli and MAC-T cells than marked signs of adhesion and invasion. Accordingly, mostly fitness traits that may also contribute to efficient colonization of the E. coli primary habitat, the gut, have been utilized by the mastitis isolates under these conditions. In this study, RNA-Seq was employed to assess the bacterial transcriptional response to milk whey. According to our transcriptome data, the lack of positively deregulated and also of true virulence-associated determinants in both of the mastitis isolates indicated that E. coli might have adapted by other means to the udder (or at least mammary secretion) as an inflammatory site. We identified traits that promote bacterial growth and survival in milk whey. The ability to utilize citrate promotes fitness and survival of E. coli that are thriving in mammary secretions. According to our results, lactoferrin has only weak impact on E. coli in mammary secretions. At the same time bacterial determinants involved in iron assimilation were negatively regulated, suggesting that, at least during the first hours, iron assimilation is not a challenge to E. coli colonizing the mammary gland. It has been hypothesized that cellular iron stores cause temporary independency to extracellular accessible iron. According to our transcriptome data, this hypothesis was supported and places iron uptake systems beyond the speculative importance that has been suggested before, at least during early phases of infection. It has also been shown that the ability to resist extracytoplasmic stress, by oxidative conditions as well as host defensins, is of substantial importance for bacterial survival in mammary secretions. In summary, the presented thesis addresses important aspects of host-pathogen interaction and bacterial conversion to hostile conditions during colonization of the mastitis inflammatory site, the mammary gland. N2 - Bei der bakteriellen Mastitis handelt es sich um eine Infektion der bovinen Milchdrüse, ausgelöst durch Eintritt und Wachstum der Bakterien im Euter der Kuh. Krankheitsverlauf und Ursache werden beeinflusst durch Umweltfaktoren, das Immunsystem des Wirtes und die Eigenschaften des bakteriellen Erregers. Die Spezies Escherichia coli (E. coli) ist einer der häufigsten Erreger der akuten bovinen Mastitis. Generell können pathogene E. coli -Stämme entsprechend ihres Infektionsortes in verschiedene Pathotypen klassifiziert werden, die durch eine individuelle Kombination verschiedener Virulenzfaktoren gekennzeichnet sind. Eine eindeutige Unterscheidung von E. coli –Mastitiserregern und kommensalen E. coli -Stämmen ist bisher nicht beschrieben. Diese Studie befasst sich mit der Charakterisierung virulenz-assozierter Eigenschaften von E. coli –Isolaten der bovinen Mastitis. Dazu wurden Untersuchungen unter Bedingungen durchgeführt, die denen während der Anfangsphase der Mastitis entsprechen. Die Virulenz und Fitness-assoziierten Eigenschaften ausgewählter E. coli Mastitis- und Fäkalisolate wurden in vergleichenden phenotypischen Assays identifiziert und analysiert. Zur Untersuchung der bakteriellen Fitness in Milchdrüsensekreten wurde native Molke mit antimikrobiellen Eigenschaften von Rohmilch genutzt. Dabei stellte sich heraus dass E. coli Fäkalisolate eine heterogene Gruppe bilden. Innerhalb dieser Gruppe wiesen einige Isolate eine verminderte Überlebensrate auf. Andere Fäkalisolate zeigten eine höhere Überlebensrate, ähnlich der Überlebensrate von Mastitiserregern. Im Gegensatz zum ihrem grundsätzlich guten Überleben in Molke zeigten Mastitisisolate keine einheitlichen phänotypischen Merkmale bei Wachstum mit 1) Lactose als einziger Kohlenstoffquelle, 2) Eisenlimitierung, oder 3) unter Einfluss von lingualem antimikrobiellem Peptid (LAP), einem bedeutenden Defensin der Wirtsantwort im Euter. Die verminderte Fähigkeit in Milchdrüsensekreten zu überleben korrelierte mit der konzentrationsabhängigen Überlebensfähigkeit in Gegenwart von LAP. Dies lässt vermuten dass eine Anpassung der Bakterien an die Lebensbedingungen in der bovinen Milchdrüse der Resistenz gegenüber Defensinen (u.a. LAP) bedarf. Den Mastitis-isolaten E. coli 1303 und ECC 1470 fehlen diverse Virulenzgene die bereits mit Mastitis assoziiert werden konnten. Um zu bestimmen ob Unterschiede in der Genexpression beider E. coli Isolate dazu beitragen Mastitis auszulösen, wurden Transkriptomanalysen durchgeführt. Dabei sollten vor allem Kandidatengene bestimmt werden, die an der Wirt-Pathogen-Interaktion beteiligt sind oder zur bakteriellen Fitness oder Virulenz der Erreger beitragen. Auf der Basis von DNA Microarrays wurde die Genexpression von E. coli 1303 und ECC 1470 in Gegenwart von immortalisierten Zellen des bovinen Milchdrüsenepithels (MAC-T) bestimmt. Zusätzlich wurde die Fähigkeit zur Zelladhäsion und Internalisierung beider Isolate untersucht. Die bakterielle Transkriptionsantwort in Gegenwart der Wirtszellen ergab, dass Erreger und Wirtszellen eher um den Bedarf an Nährstoffen und Sauerstoff konkurrierten, anstatt deutliche Anzeichen der Zelladhäsion oder Invasion zu zeigen. Beide Isolate nutzten vornehmlich Fitnesseigenschaften, die auch bei der Besiedlung des Darms als dem primären Habitat von E. coli verwendet werden. In dieser Studie wurde außerdem die Genexpression von E. coli 1303 und ECC 1470 in Reaktion auf Molke aus Rohmilch mittels Gesamt-Transkriptom-Sequenzierung (RNA Seq) untersucht. Die Transkriptomanalyse ergab keine wirklich deregulierten virulenz-assozierten Gene in einer der beiden E. coli Mastitis Isolate. Ferner konnten Eigenschaften identifiziert werden, die zum Wachstum und Überleben in nativer Molke beitragen. Die Fähigkeit, Citrat zu verwerten, begünstigt das erfolgreiche Überleben in Milchdrüsensekten und stellt einen wichtigen Fitnessfaktor dar. Unsere Transkriptomdaten bestätigen dass Lactoferrin nur geringen Einfluss auf das Wachstum, von E. coli in Milchdrüsensekreten, hat. Die Expression bakterieller Determinanten, die an der Aufnahme von Eisen beteiligt sind, wurde herunterreguliert. Dies lässt darauf schließen dass Eisenaufnahme in den ersten Stunden der Kolonisierung durch die Erreger keine essentielle Fitnesseigenschaft darstellt. Vermutlich reicht die intrazelluläre Menge an Eisen aus, um eine zeitweise Unabhängigkeit von extrazellular verfügbarem Eisen zu ermöglichen. Diese These konnte durch unsere Transkriptomdaten gestützt werden und stellt eine wichtige Entdeckung in Bezug auf die Verfügbarkeit von Eisen während der Kolonisierung der Milchdrüse dar. Unsere Daten zeigen, dass die Resistenz gegenüber extrazellulärem Stress durch oxidative Bedingungen und Defensine des Wirtes von großer Bedeutung für das bakterielle Überleben in Milchdrüsensekreten ist. Die vorliegende Thesis befasst sich mit wichtigen Aspekten der Wirt-Pathogen-Interaktion und der Anpassung an die antimikrobiellen Bedingungen während der Kolonisierung der Milchdrüse als Ort der Infektion. KW - Escherichia coli KW - mastitis KW - infection KW - virulence KW - transcriptome analysis KW - Kuh KW - Brustdrüsenentzündung KW - Virulenz Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-100934 ER - TY - JOUR A1 - Wu, Hao A1 - Zhao, Xiufeng A1 - Hochrein, Sophia M. A1 - Eckstein, Miriam A1 - Gubert, Gabriela F. A1 - Knöpper, Konrad A1 - Mansilla, Ana Maria A1 - Öner, Arman A1 - Doucet-Ladevèze, Remi A1 - Schmitz, Werner A1 - Ghesquière, Bart A1 - Theurich, Sebastian A1 - Dudek, Jan A1 - Gasteiger, Georg A1 - Zernecke, Alma A1 - Kobold, Sebastian A1 - Kastenmüller, Wolfgang A1 - Vaeth, Martin T1 - Mitochondrial dysfunction promotes the transition of precursor to terminally exhausted T cells through HIF-1α-mediated glycolytic reprogramming JF - Nature Communications N2 - T cell exhaustion is a hallmark of cancer and persistent infections, marked by inhibitory receptor upregulation, diminished cytokine secretion, and impaired cytolytic activity. Terminally exhausted T cells are steadily replenished by a precursor population (Tpex), but the metabolic principles governing Tpex maintenance and the regulatory circuits that control their exhaustion remain incompletely understood. Using a combination of gene-deficient mice, single-cell transcriptomics, and metabolomic analyses, we show that mitochondrial insufficiency is a cell-intrinsic trigger that initiates the functional exhaustion of T cells. At the molecular level, we find that mitochondrial dysfunction causes redox stress, which inhibits the proteasomal degradation of hypoxia-inducible factor 1α (HIF-1α) and promotes the transcriptional and metabolic reprogramming of Tpex cells into terminally exhausted T cells. Our findings also bear clinical significance, as metabolic engineering of chimeric antigen receptor (CAR) T cells is a promising strategy to enhance the stemness and functionality of Tpex cells for cancer immunotherapy. KW - cytotoxic T cells KW - infection KW - lymphocyte differentiation KW - translational research Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-358052 VL - 14 ER - TY - JOUR A1 - Wu, Hao A1 - Reimann, Sabine A1 - Siddiqui, Sophiya A1 - Haag, Rainer A1 - Siegmund, Britta A1 - Dernedde, Jens A1 - Glauben, Rainer T1 - dPGS Regulates the Phenotype of Macrophages via Metabolic Switching JF - Macromolecular Bioscience N2 - The synthetic compound dendritic polyglycerol sulfate (dPGS) is a pleiotropic acting molecule but shows a high binding affinity to immunological active molecules as L‐/P‐selectin or complement proteins leading to well described anti‐inflammatory properties in various mouse models. In order to make a comprehensive evaluation of the direct effect on the innate immune system, macrophage polarization is analyzed in the presence of dPGS on a phenotypic but also metabolic level. dPGS administered macrophages show a significant increase of MCP1 production paralleled by a reduction of IL‐10 secretion. Metabolic analysis reveals that dPGS could potently enhance the glycolysis and mitochondrial respiration in M0 macrophages as well as decrease the mitochondrial respiration of M2 macrophages. In summary the data indicate that dPGS polarizes macrophages into a pro‐inflammatory phenotype in a metabolic pathway‐dependent manner. KW - infection KW - macrophage polarization KW - MCP1 KW - metabolic switch KW - polyglycerol sulfates Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-212711 VL - 19 IS - 12 ER - TY - JOUR A1 - Unnewehr, Markus A1 - Stich, August T1 - Fighting Hepatitis B in North Korea: Feasibility of a Bi-modal Prevention Strategy JF - Journal of Korean Medical Science N2 - In North Korea, the prevalence of hepatitis B is high due to natural factors, gaps in vaccination, and the lack of antiviral treatment. Aid projects are urgently needed, however impeded by North Korea's political and economical situation and isolation. The feasibility of a joint North Korean and German humanitarian hepatitis B prevention program was assessed. Part 1: Hepatitis B vaccination catch-up campaign. Part 2: Implementation of endoscopic ligation of esophageal varices (EVL) by trainings in Germany and North Korea. By vaccinating 7 million children between 2010 and 2012, the hepatitis B vaccination gap was closed. Coverage of 99.23% was reached. A total of 11 hepatitis B-induced liver cirrhosis patients (mean age 41.1 yr) with severe esophageal varices and previous bleedings were successfully treated by EVL without major complications. A clinical standard operating procedure, a feedback system and a follow-up plan were developed. The bi-modal preventive strategy was implemented successfully. Parts of the project can serve as an example for other low-income countries, however its general transferability is limited due to the special circumstances in North Korea. KW - therapy KW - hepatitis B KW - Democratic People's Republic of Korea KW - ligation KW - cirrhosis KW - diagnosis KW - infection KW - health KW - primary prophylaxis KW - mass vaccination KW - esophagoscopy Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-138773 VL - 30 ER - TY - JOUR A1 - Twisselmann, Nele A1 - Pagel, Julia A1 - Künstner, Axel A1 - Weckmann, Markus A1 - Hartz, Annika A1 - Glaser, Kirsten A1 - Hilgendorff, Anne A1 - Göpel, Wolfgang A1 - Busch, Hauke A1 - Herting, Egbert A1 - Weinberg, Jason B. A1 - Härtel, Christoph T1 - Hyperoxia/Hypoxia Exposure Primes a Sustained Pro-Inflammatory Profile of Preterm Infant Macrophages Upon LPS Stimulation JF - Frontiers in Immunology N2 - Preterm infants are highly susceptible to sustained lung inflammation, which may be triggered by exposure to multiple environmental cues such as supplemental oxygen (O\(_2\)) and infections. We hypothesized that dysregulated macrophage (MФ) activation is a key feature leading to inflammation-mediated development of bronchopulmonary dysplasia (BPD) in preterm infants. Therefore, we aimed to determine age-dependent differences in immune responses of monocyte-derived MФ comparing cord blood samples derived from preterm (n=14) and term (n=19) infants as well as peripheral blood samples from healthy adults (n=17) after lipopolysaccharide (LPS) exposure. Compared to term and adult MФ, LPS-stimulated preterm MФ showed an enhanced and sustained pro-inflammatory immune response determined by transcriptome analysis, cytokine release inducing a RORC upregulation due to T cell polarization of neonatal T cells, and TLR4 surface expression. In addition, a double-hit model was developed to study pulmonary relevant exposure factors by priming MФ with hyperoxia (O\(_2\) = 65%) or hypoxia (O\(_2\) = 3%) followed by lipopolysaccharide (LPS, 100ng/ml). When primed by 65% O\(_2\), subsequent LPS stimulation in preterm MФ led to an exaggerated pro-inflammatory response (e.g. increased HLA-DR expression and cytokine release) compared to LPS stimulation alone. Both, exposure to 65% or 3% O\(_2\) together with subsequent LPS stimulation, resulted in an exaggerated pro-inflammatory response of preterm MФ determined by transcriptome analysis. Downregulation of two major transcriptional factors, early growth response gene (Egr)-2 and growth factor independence 1 (Gfi1), were identified to play a role in the exaggerated pro-inflammatory response of preterm MФ to LPS insult after priming with 65% or 3% O\(_2\). Preterm MФ responses to LPS and hyperoxia/hypoxia suggest their involvement in excessive inflammation due to age-dependent differences, potentially mediated by downregulation of Egr2 and Gfi1 in the developing lung. KW - preterm infants KW - sustained inflammation KW - macrophages KW - hyperoxia KW - hypoxia KW - infection KW - bronchopulmonary dysplasia Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-250356 SN - 1664-3224 VL - 12 ER - TY - JOUR A1 - Tappe, D. A1 - Stich, A. A1 - Langeheinecke, A. A1 - von Sonnenburg, F. A1 - Muntau, B. A1 - Schaefer, J. A1 - Slesak, G. T1 - Suspected new wave of muscular sarcocystosis in travellers returning from Tioman Island, Malaysia, May 2014 JF - Eurosurveillance N2 - In May 2014, six patients presented in Germany with a Sarcocystis-associated febrile myositis syndrome after returning from Tioman Island, Malaysia. During two earlier waves of infections, in 2011 and 2012, about 100 travellers returning to various European countries from the island were affected. While the first two waves were associated with travel to Tioman Island mostly during the summer months, this current series of infections is associated with travel in early spring, possibly indicating an upcoming new epidemic. KW - infection KW - nesbitti KW - outbreak Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-116195 VL - 19 IS - 21 ER - TY - JOUR A1 - Strobel, Lea A1 - Johswich, Kay O. T1 - Anticoagulants impact on innate immune responses and bacterial survival in whole blood models of Neisseria meningitidis infection JF - Scientific Reports N2 - Neisseria meningitidis (meningococcus) causes invasive diseases such as meningitis or septicaemia. Ex vivo infection of human whole blood is a valuable tool to study meningococcal virulence factors and the host innate immune responses. In order to consider effects of cellular mediators, the coagulation cascade must be inhibited to avoid clotting. There is considerable variation in the anticoagulants used among studies of N. meningitidis whole blood infections, featuring citrate, heparin or derivatives of hirudin, a polypeptide from leech saliva. Here, we compare the influence of these three different anticoagulants, and additionally Mg/EGTA, on host innate immune responses as well as on viability of N. meningitidis strains isolated from healthy carriers and disease cases, reflecting different sequence types and capsule phenotypes. We found that the anticoagulants significantly impact on cellular responses and, strain-dependently, also on bacterial survival. Hirudin does not inhibit complement and is therefore superior over the other anticoagulants; indeed hirudin-plasma most closely reflects the characteristics of serum during N. meningitidis infection. We further demonstrate the impact of heparin on complement activation on N. meningitidis and its consequences on meningococcal survival in immune sera, which appears to be independent of the heparin binding antigens Opc and NHBA. KW - infection KW - pathogens KW - Neisseria meningitidis KW - anticoagulants Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-176226 VL - 8 IS - 10225 ER - TY - JOUR A1 - Srivastava, Mugdha A1 - Bencurova, Elena A1 - Gupta, Shishir K. A1 - Weiss, Esther A1 - Löffler, Jürgen A1 - Dandekar, Thomas T1 - Aspergillus fumigatus challenged by human dendritic cells: metabolic and regulatory pathway responses testify a tight battle JF - Frontiers in Cellular and Infection Microbiology N2 - Dendritic cells (DCs) are antigen presenting cells which serve as a passage between the innate and the acquired immunity. Aspergillosis is a major lethal condition in immunocompromised patients caused by the adaptable saprophytic fungus Aspergillus fumigatus. The healthy human immune system is capable to ward off A. fumigatus infections however immune-deficient patients are highly vulnerable to invasive aspergillosis. A. fumigatus can persist during infection due to its ability to survive the immune response of human DCs. Therefore, the study of the metabolism specific to the context of infection may allow us to gain insight into the adaptation strategies of both the pathogen and the immune cells. We established a metabolic model of A. fumigatus central metabolism during infection of DCs and calculated the metabolic pathway (elementary modes; EMs). Transcriptome data were used to identify pathways activated when A. fumigatus is challenged with DCs. In particular, amino acid metabolic pathways, alternative carbon metabolic pathways and stress regulating enzymes were found to be active. Metabolic flux modeling identified further active enzymes such as alcohol dehydrogenase, inositol oxygenase and GTP cyclohydrolase participating in different stress responses in A. fumigatus. These were further validated by qRT-PCR from RNA extracted under these different conditions. For DCs, we outlined the activation of metabolic pathways in response to the confrontation with A. fumigatus. We found the fatty acid metabolism plays a crucial role, along with other metabolic changes. The gene expression data and their analysis illuminate additional regulatory pathways activated in the DCs apart from interleukin regulation. In particular, Toll-like receptor signaling, NOD-like receptor signaling and RIG-I-like receptor signaling were active pathways. Moreover, we identified subnetworks and several novel key regulators such as UBC, EGFR, and CUL3 of DCs to be activated in response to A. fumigatus. In conclusion, we analyze the metabolic and regulatory responses of A. fumigatus and DCs when confronted with each other. KW - infection KW - dendritic cells KW - Aspergillus fumigalus KW - metabolic modelling KW - signalling Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201368 VL - 9 ER - TY - JOUR A1 - Sporbert, Anje A1 - Cseresnyes, Zoltan A1 - Heidbreder, Meike A1 - Domaing, Petra A1 - Hauser, Stefan A1 - Kaltschmidt, Barbara A1 - Kaltschmidt, Christian A1 - Heilemann, Mike A1 - Widera, Darius T1 - Simple Method for Sub-Diffraction Resolution Imaging of Cellular Structures on Standard Confocal Microscopes by Three-Photon Absorption of Quantum Dots JF - PLoS ONE N2 - This study describes a simple technique that improves a recently developed 3D sub-diffraction imaging method based on three-photon absorption of commercially available quantum dots. The method combines imaging of biological samples via tri-exciton generation in quantum dots with deconvolution and spectral multiplexing, resulting in a novel approach for multi-color imaging of even thick biological samples at a 1.4 to 1.9-fold better spatial resolution. This approach is realized on a conventional confocal microscope equipped with standard continuous-wave lasers. We demonstrate the potential of multi-color tri-exciton imaging of quantum dots combined with deconvolution on viral vesicles in lentivirally transduced cells as well as intermediate filaments in three-dimensional clusters of mouse-derived neural stem cells (neurospheres) and dense microtubuli arrays in myotubes formed by stacks of differentiated C2C12 myoblasts. KW - HIV KW - stem-cell KW - infection Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130963 VL - 8 IS - 5 ER - TY - JOUR A1 - Smith, Craig J. A1 - Bray, Benjamin D. A1 - Hoffman, Alex A1 - Meisel, Andreas A1 - Heuschmann, Peter U. A1 - Wolfe, Charles D. A. A1 - Tyrrell, Pippa J. A1 - Rudd, Anthony G. T1 - Can a novel clinical risk score improve pneumonia prediction in acute stroke care? A UK multicenter cohort study JF - Journal of the American Heart Association N2 - Background Pneumonia frequently complicates stroke and has amajor impact on outcome. We derived and internally validated a simple clinical risk score for predicting stroke-associated pneumonia (SAP), and compared the performance with an existing score (A\(^{2}\)DS\(^{2}\)). Methods and Results We extracted data for patients with ischemic stroke or intracerebral hemorrhage from the Sentinel Stroke National Audit Programme multicenter UK registry. The data were randomly allocated into derivation (n=11 551) and validation (n=11 648) samples. A multivariable logistic regression model was fitted to the derivation data to predict SAP in the first 7 days of admission. The characteristics of the score were evaluated using receiver operating characteristics (discrimination) and by plotting predicted versus observed SAP frequency in deciles of risk (calibration). Prevalence of SAP was 6.7% overall. The final 22-point score (ISAN: prestroke Independence [modified Rankin scale], Sex, Age, National Institutes of Health Stroke Scale) exhibited good discrimination in the ischemic stroke derivation (C-statistic 0.79; 95% CI 0.77 to 0.81) and validation (C-statistic 0.78; 95% CI 0.76 to 0.80) samples. It was well calibrated in ischemic stroke and was further classified into meaningful risk groups (low 0 to 5, medium6 to 10, high 11 to 14, and very high >= 15) associated with SAP frequencies of 1.6%, 4.9%, 12.6%, and 26.4%, respectively, in the validation sample. Discrimination for both scores was similar, although they performed less well in the intracerebral hemorrhage patients with an apparent ceiling effect. Conclusions The ISAN score is a simple tool for predicting SAP in clinical practice. External validation is required in ischemic and hemorrhagic stroke cohorts. KW - acute ischemic stroke KW - medical complications KW - infection KW - diagnosis KW - stroke-associated pneumonia KW - clinical risk score KW - pneumonia KW - stroke, acute KW - metaanalysis KW - reliability KW - dysphagia KW - scale KW - mortality KW - intracerebral hemorrhage Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144602 VL - 4 IS - 1 ER - TY - JOUR A1 - Rose, Markus A. A1 - Damm, Oliver A1 - Greiner, Wolfgang A1 - Knuf, Markus A1 - Wutzler, Peter A1 - Liese, Johannes G. A1 - Krüger, Hagen A1 - Wahn, Ulrich A1 - Schaberg, Tom A1 - Schwehm, Markus A1 - Kochmann, Thomas F. A1 - Eichner, Martin T1 - The epidemiological impact of childhood influenza vaccination using live-attenuated influenza vaccine (LAIV) in Germany: predictions of a simulation study JF - BMC Infectious Diseases N2 - Background: Routine annual influenza vaccination is primarily recommended for all persons aged 60 and above and for people with underlying chronic conditions in Germany. Other countries have already adopted additional childhood influenza immunisation programmes. The objective of this study is to determine the potential epidemiological impact of implementing paediatric influenza vaccination using intranasally administered live-attenuated influenza vaccine (LAIV) in Germany. Methods: A deterministic age-structured model is used to simulate the population-level impact of different vaccination strategies on the transmission dynamics of seasonal influenza in Germany. In our base-case analysis, we estimate the effects of adding a LAIV-based immunisation programme targeting children 2 to 17 years of age to the existing influenza vaccination policy. The data used in the model is based on published evidence complemented by expert opinion. Results: In our model, additional vaccination of children 2 to 17 years of age with LAIV leads to the prevention of 23.9 million influenza infections and nearly 16 million symptomatic influenza cases within 10 years. This reduction in burden of disease is not restricted to children. About one third of all adult cases can indirectly be prevented by LAIV immunisation of children. Conclusions: Our results demonstrate that vaccinating children 2-17 years of age is likely associated with a significant reduction in the burden of paediatric influenza. Furthermore, annual routine childhood vaccination against seasonal influenza is expected to decrease the incidence of influenza among adults and older people due to indirect effects of herd protection. In summary, our model provides data supporting the introduction of a paediatric influenza immunisation programme in Germany. KW - vaccination KW - live-attenuated influenza vaccine KW - children KW - transmission model KW - metanalysis KW - recommendations KW - hospitalizations KW - burden KW - infection KW - young children KW - seasonal influenza KW - United States KW - disease KW - efficacy KW - Germany KW - influenza Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117563 SN - 1471-2334 VL - 14 IS - 40 ER - TY - JOUR A1 - Roesler, Joachim A1 - Segerer, Florian A1 - Morbach, Henner A1 - Kleinert, Stefan A1 - Thieme, Sebastian A1 - Rösen-Wolff, Angela A1 - Liese, Johannes G. T1 - P67-phox (NCF2) Lacking Exons 11 and 12 Is Functionally Active and Leads to an Extremely Late Diagnosis of Chronic Granulomatous Disease (CGD) JF - PLoS One N2 - Two brothers in their fifties presented with a medical history of suspected fungal allergy, allergic bronchopulmonary aspergillosis, alveolitis, and invasive aspergillosis and pulmonary fistula, respectively. Eventually, after a delay of 50 years, chronic granulomatous disease (CGD) was diagnosed in the index patient. We found a new splice mutation in the NCF2 (p67-phox) gene, c.1000+2T -> G, that led to several splice products one of which lacked exons 11 and 12. This deletion was in frame and allowed for remarkable residual NADPH oxidase activity as determined by transduction experiments using a retroviral vector. We conclude that p67-phox which lacks the 34 amino acids encoded by the two exons can still exert considerable functional activity. This activity can partially explain the long-term survival of the patients without adequate diagnosis and treatment, but could not prevent progressing lung damage. KW - P67(PHOX) KW - NADPH oxidase KW - European experience KW - interferon gamma KW - gene KW - region KW - prophylaxis KW - infection KW - mutation Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-134948 VL - 7 IS - 4 ER - TY - JOUR A1 - Rieger, C. T. A1 - Liss, B. A1 - Mellinghoff, S. A1 - Buchheidt, D. A1 - Cornely, O. A. A1 - Egerer, G. A1 - Heinz, W. J. A1 - Hentrich, M. A1 - Maschmeyer, G. A1 - Mayer, K. A1 - Sandherr, M. A1 - Silling, G. A1 - Ullmann, A. A1 - Vehreschild, M. J. G. T. A1 - von Lilienfeld-Toal, M. A1 - Wolf, H. H. A1 - Lehners, N. T1 - Anti-infective vaccination strategies in patients with hematologic malignancies or solid tumors-Guideline of the Infectious Diseases Working Party (AGIHO) of the German Society for Hematology and Medical Oncology (DGHO) JF - Annals of Oncology N2 - Infectious complications are a significant cause of morbidity and mortality in patients with malignancies specifically when receiving anticancer treatments. Prevention of infection through vaccines is an important aspect of clinical care of cancer patients. Immunocompromising effects of the underlying disease as well as of antineoplastic therapies need to be considered when devising vaccination strategies. This guideline provides clinical recommendations on vaccine use in cancer patients including autologous stem cell transplant recipients, while allogeneic stem cell transplantation is subject of a separate guideline. The document was prepared by the Infectious Diseases Working Party (AGIHO) of the German Society for Hematology and Medical Oncology (DGHO) by reviewing currently available data and applying evidence-based medicine criteria. KW - infection KW - anti-infective vaccination KW - cancer KW - immunosuppression KW - autologous stem cell transplantation Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-226196 VL - 29 IS - 6 ER - TY - THES A1 - Pekárek, Lukáš T1 - Single-Molecule Approaches To Study Frameshifting Mechanisms T1 - Einzelmolekülansätze zur Untersuchung von Frameshifting-Mechanismen N2 - The RNAs of many viruses contain a frameshift stimulatory element (FSE) that grants access to an alternate reading frame via −1 programmed ribosomal frameshifting (PRF). This −1PRF is essential for effective viral replication. The −1PRF efficiency relies on the presence of conserved RNA elements within the FSE, such as a slippery sequence, spacer, and a downstream secondary structure – often a hairpin or a pseudoknot. The PRF efficiency is also affected by trans-acting factors such as proteins, miRNAs and metabolites. The interactions of these factors with the RNA and the translation machinery have not yet been completely understood. Traditional ensemble methods used previously to study these events focus on the whole population of molecular species. This results in innate averaging of the molecular behavior and a loss of heterogeneity information. Here, we first established the experimental workflow to study the RNA structures and the effect of potential trans-acting factors using single-molecule force spectroscopy technique, optical tweezers. Additionally, to streamline the data analysis, we developed an algorithm for automatized data processing. Next, we harnessed this knowledge to study viral RNA elements responsible for stimulation of PRF and how the presence of trans-acting factors affects the RNA behavior. We further complemented these single-molecule structural data with ensemble functional assays to gain a complex view on the dynamics behind the programmed ribosomal frameshifting. Specifically, two different viral RNA elements have been studied in the presented work. First, the dynamics of SARS-CoV-2 FSE and the role of extended sequences have been explored. Then, the mode of action of the host-encoded trans-acting factor ZAP-S inhibition of SARS-CoV-2 PRF has been examined. Finally, the mechanism of the trans-acting viral factor induced PRF in Encephalomyocarditis virus (EMCV) has been uncovered. N2 - Die RNAs vieler Viren enthalten ein Lese-Rasterverschiebung-stimulierendes Element (FSE), das über die −1 programmierte ribosomale Rasterverschiebung (PRF) Zugriff auf einen alternativen Leserahmen gewährt. Dieser −1PRF ist für eine effektive Virusreplikation unerlässlich. Die −1PRF-Effizienz beruht auf dem Vorhandensein konservierter RNA-Elemente innerhalb des FSE, wie z.B. einer Slippery-Sequenz, einem Platzhalter und einer nachgelagerten Sekundärstruktur – oft eine Haarnadel oder ein Pseudoknoten. Die −1PRF-Effizienz wird auch durch trans-aktive Faktoren wie Proteine, miRNAs und Metaboliten beeinflusst. Die Wechselwirkungen dieser Faktoren mit der RNA und der Translationsmaschinerie sind noch nicht vollständig verstanden. Traditionelle Ensemble-Methoden, die früher zur Untersuchung dieser Ereignisse verwendet wurden, konzentrieren sich auf die gesamte Population molekularer Spezies. Dies führt zu einer inhärenten Durchschnittsbildung des molekularen Verhaltens und einem Verlust von Heterogenitätsinformationen. Hier haben wir zunächst den experimentellen Arbeitsablauf zur Untersuchung der RNA-Strukturen und der Wirkung potenzieller trans-aktiver Faktoren mithilfe der Einzelmolekül-Kraftspektroskopietechnik Optischer Pinzetten etabliert. Um die Datenanalyse zu optimieren, haben wir außerdem einen Algorithmus zur automatisierten Datenverarbeitung entwickelt. Als nächstes nutzten wir dieses Wissen, um virale RNA-Elemente zu untersuchen, die für die Stimulierung von −1PRF verantwortlich sind, und wie sich das Vorhandensein trans-aktiver Faktoren auf das Verhalten der RNA auswirkt. Wir haben diese Einzelmolekülstrukturdaten weiter durch Ensemble-Funktionsassays ergänzt, um einen komplexen Überblick über die Dynamik hinter der programmierten ribosomalen Rasterverschiebung zu erhalten. Konkret wurden in der vorgestellten Arbeit zwei verschiedene virale RNA-Elemente untersucht. Zunächst wurden die Dynamik des SARS-CoV-2-FSE und die Rolle erweiterter Sequenzen untersucht. Anschließend wurde die hemmende Wirkungsweise des vom Wirt kodierten trans-wirkenden Faktors ZAP-S auf SARS-CoV-2-PRF untersucht. Schließlich wurde der Mechanismus der, durch den trans-aktiven Virusfaktor induzierten PRF beim Enzephalomyokarditis-Virus (EMCV), entschlüsselt KW - translation KW - infection KW - frameshifting Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-346112 ER - TY - JOUR A1 - Okoro, Chinyere K. A1 - Barquist, Lars A1 - Connor, Thomas R. A1 - Harris, Simon R. A1 - Clare, Simon A1 - Stevens, Mark P. A1 - Arends, Mark J. A1 - Hale, Christine A1 - Kane, Leanne A1 - Pickard, Derek J. A1 - Hill, Jennifer A1 - Harcourt, Katherine A1 - Parkhill, Julian A1 - Dougan, Gordon A1 - Kingsley, Robert A. T1 - Signatures of adaptation in human invasive Salmonella Typhimurium ST313 populations from sub-Saharan Africa JF - PLoS Neglected Tropical Diseases N2 - Two lineages of Salmonella enterica serovar Typhimurium (S. Typhimurium) of multi-locus sequence type ST313 have been linked with the emergence of invasive Salmonella disease across sub-Saharan Africa. The expansion of these lineages has a temporal association with the HIV pandemic and antibiotic usage. We analysed the whole genome sequence of 129 ST313 isolates representative of the two lineages and found evidence of lineage-specific genome degradation, with some similarities to that observed in S. Typhi. Individual ST313 S. Typhimurium isolates exhibit a distinct metabolic signature and modified enteropathogenesis in both a murine and cattle model of colitis, compared to S. Typhimurium outside of the ST313 lineages. These data define phenotypes that distinguish ST313 isolates from other S. Typhimurium and may represent adaptation to a distinct pathogenesis and lifestyle linked to an-immuno-compromised human population. KW - genome sequence KW - infection KW - pathogenicity KW - children KW - disease KW - adults KW - identification KW - Escherichia coli KW - virulence Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143779 VL - 9 IS - 3 ER - TY - JOUR A1 - Ntoukas, Vasileios A1 - Tappe, Dennis A1 - Pfütze, Daniel A1 - Simon, Michaela A1 - Holzmann, Thomas T1 - Cerebellar Cysticercosis Caused by Larval Taenia crassiceps Tapeworm in Immunocompetent Woman, Germany JF - Emerging Infectious Diseases N2 - Human cysticercosis caused by Taenia crassiceps tapeworm larvae involves the muscles and subcutis mostly in immunocompromised patients and the eye in immunocompetent persons. We report a successfully treated cerebellar infection in an immunocompetent woman. We developed serologic tests, and the parasite was identified by histologic examination and 12s rDNA PCR and sequencing. KW - helminths KW - acquired immunodeficiency-syndrome KW - foxes vulpes-vulpes KW - invasive cysticerocsis KW - infection KW - patient KW - AIDS Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-131686 N1 - Public Domain VL - 19 IS - 12 ER - TY - JOUR A1 - Nadella, Vinod A1 - Mohanty, Aparna A1 - Sharma, Lalita A1 - Yellaboina, Sailu A1 - Mollenkopf, Hans-Joachim A1 - Mazumdar, Varadendra Balaji A1 - Palaparthi, Ramesh A1 - Mylavarapu, Madhavi B. A1 - Maurya, Radheshyam A1 - Kurukuti, Sreenivasulu A1 - Rudel, Thomas A1 - Prakash, Hridayesh T1 - Inhibitors of Apoptosis Protein Antagonists (Smac Mimetic Compounds) Control Polarization of Macrophages during Microbial Challenge and Sterile Inflammatory Responses JF - Frontiers in Immunology N2 - Apoptosis is a physiological cell death process essential for development, tissue homeostasis, and for immune defense of multicellular animals. Inhibitors of apoptosis proteins (IAPs) regulate apoptosis in response to various cellular assaults. Using both genetic and pharmacological approaches we demonstrate here that the IAPs not only support opportunistic survival of intracellular human pathogens like Chlamydia pneumoniae but also control plasticity of iNOS+ M1 macrophage during the course of infection and render them refractory for immune stimulation. Treatment of Th1 primed macrophages with birinapant (IAP-specific antagonist) inhibited NO generation and relevant proteins involved in innate immune signaling. Accordingly, birinapant promoted hypoxia, angiogenesis, and tumor-induced M2 polarization of iNOS+ M1 macrophages. Interestingly, birinapant-driven changes in immune signaling were accompanied with changes in the expression of various proteins involved in the metabolism, and thus revealing the new role of IAPs in immune metabolic reprogramming in committed macrophages. Taken together, our study reveals the significance of IAP targeting approaches (Smac mimetic compounds) for the management of infectious and inflammatory diseases relying on macrophage plasticity. KW - apoptosis KW - macrophages immunobiology KW - inflammation mediators KW - polarization KW - infection KW - hypothalamus Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-197484 SN - 1664-3224 VL - 8 IS - 1792 ER - TY - JOUR A1 - Milanez-Almeida, P. A1 - Ulas, T. A1 - Pasztoi, M. A1 - Glage, S. A1 - Schughart, K. A1 - Lutz, M. B. A1 - Schultze, J. L. A1 - Huehn, J. T1 - CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells with suppressive activity towards T cells accumulate in lungs of influenza A virus-infected mice JF - European Journal of Microbiology and Immunology N2 - Influenza A virus (IAV) infection causes an acute respiratory disease characterized by a strong inflammatory immune response and severe immunopathology. Proinflammatory mechanisms are well described in the murine IAV infection model, but less is known about the mechanisms leading to the resolution of inflammation. Here, we analyzed the contribution of CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells to this process. An accumulation of CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells within the lungs was observed during the course of IAV infection. Phenotypic characterization of these CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells by flow cytometry and RNA-Seq revealed an activated phenotype showing both pro- and anti-inflammatory features, including the expression of inducible nitric oxide synthase (iNOS) by a fraction of cells in an IFN-γ-dependent manner. Moreover, CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells isolated from lungs of IAV-infected animals displayed suppressive activity when tested in vitro, and iNOS inhibitors could abrogate this suppressive activity. Collectively, our data suggest that during IAV infection, CD11b\(^{+}\)Ly6C\(^{++}\)Ly6G\(^{-}\) cells acquire immunoregulatory function, which might contribute to the prevention of pathology during this life-threatening disease. KW - monocytes KW - inducible nitric oxide synthase KW - influenza A virus KW - infection KW - immuno suppression Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-149583 VL - 5 IS - 4 ER - TY - JOUR A1 - Makgotlho, Phuti E. A1 - Marincola, Gabriella A1 - Schäfer, Daniel A1 - Liu, Quian A1 - Bae, Taeok A1 - Geiger, Tobias A1 - Wasserman, Elizabeth A1 - Wolz, Christine A1 - Ziebuhr, Wilma A1 - Sinha, Bhanu T1 - SDS Interferes with SaeS Signaling of Staphylococcus aureus Independently of SaePQ JF - PLOS ONE N2 - The Staphylococcus aureus regulatory saePQRS system controls the expression of numerous virulence factors, including extracellular adherence protein (Eap), which amongst others facilitates invasion of host cells. The saePQRS operon codes for 4 proteins: the histidine kinase SaeS, the response regulator SaeR, the lipoprotein SaeP and the transmembrane protein SaeQ. S. aureus strain Newman has a single amino acid substitution in the transmembrane domain of SaeS (L18P) which results in constitutive kinase activity. SDS was shown to be one of the signals interfering with SaeS activity leading to inhibition of the sae target gene eap in strains with SaeS(L) but causing activation in strains containing SaeS(P). Here, we analyzed the possible involvement of the SaeP protein and saePQ region in SDS-mediated sae/eap expression. We found that SaePQ is not needed for SDS-mediated SaeS signaling. Furthermore, we could show that SaeS activity is closely linked to the expression of Eap and the capacity to invade host cells in a number of clinical isolates. This suggests that SaeS activity might be directly modulated by structurally non-complex environmental signals, as SDS, which possibly altering its kinase/phosphatase activity. KW - host-cell invasion KW - 2-component system KW - strain Newman KW - allelic replacement KW - genome sequence KW - locus KW - gene KW - activation KW - expression KW - infection Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-128469 SN - 1932-6203 VL - 8 IS - 8 ER - TY - JOUR A1 - León-Calvijo, María A. A1 - Leal-Castro, Aura L. A1 - Almanzar-Reina, Giovanni A. A1 - Rosas-Pérez, Jaiver E. A1 - García-Castañeda, Javier E. A1 - Rivera-Monroy, Zuly J. T1 - Antibacterial activity of synthetic peptides derived from lactoferricin against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212 JF - BioMed Research International N2 - Peptides derived from human and bovine lactoferricin were designed, synthesized, purified, and characterized using RP-HPLC and MALDI-TOF-MS. Specific changes in the sequences were designed as (i) the incorporation of unnatural amino acids in the sequence, the (ii) reduction or (iii) elongation of the peptide chain length, and (iv) synthesis of molecules with different number of branches containing the same sequence. For each peptide, the antibacterial activity against Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212 was evaluated. Our results showed that Peptides I.2 (RWQWRWQWR) and I.4 ((RRWQWR)\(_{4}\)K\(_{2}\)Ahx\(_{2}\)C\(_{2}\)) exhibit bigger or similar activity against E. coli (MIC 4-33 μM) and E. faecalis (MIC 10-33 μM) when they were compared with lactoferricin protein (LF) and some of its derivate peptides as II.1 (FKCRRWQWRMKKLGA) and IV.1 (FKCRRWQWRMKKLGAPSITCVRRAE). It should be pointed out that Peptides I.2 and I.4, containing the RWQWR motif, are short and easy to synthesize; our results demonstrate that it is possible to design and obtain synthetic peptides that exhibit enhanced antibacterial activity using a methodology that is fast and low-cost and that allows obtaining products with a high degree of purity and high yield. KW - bovine lactoferricin KW - antimicrobial activity KW - infection KW - spectrum KW - mice KW - cells KW - inhibit KW - derivatives KW - loop region Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144591 IS - 453826 ER - TY - JOUR A1 - Kunz, Tobias C. A1 - Kozjak-Pavlovic, Vera T1 - Diverse facets of sphingolipid involvement in bacterial infections JF - Frontiers in Cell and Developmental Biology N2 - Sphingolipids are constituents of the cell membrane that perform various tasks as structural elements and signaling molecules, in addition to regulating many important cellular processes, such as apoptosis and autophagy. In recent years, it has become increasingly clear that sphingolipids and sphingolipid signaling play a vital role in infection processes. In many cases the attachment and uptake of pathogenic bacteria, as well as bacterial development and survival within the host cell depend on sphingolipids. In addition, sphingolipids can serve as antimicrobials, inhibiting bacterial growth and formation of biofilms. This review will give an overview of our current information about these various aspects of sphingolipid involvement in bacterial infections. KW - infection KW - pathogenic bacteria KW - sphingolipids KW - ceramide KW - autophagy Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-201757 VL - 7 IS - 203 ER - TY - JOUR A1 - Kincaid, Rodney P. A1 - Chen, Yating A1 - Cox, Jennifer E. A1 - Rethwilm, Axel A1 - Sullivan, Christopher S. T1 - Noncanonical MicroRNA (miRNA) Biogenesis Gives Rise to Retroviral Mimics of Lymphoproliferative and Immunosuppressive Host miRNAs JF - mBio N2 - MicroRNAs (miRNAs) play regulatory roles in diverse processes in both eukaryotic hosts and their viruses, yet fundamental questions remain about which viruses code for miRNAs and the functions that they serve. Simian foamy viruses (SFVs) of Old World monkeys and apes can zoonotically infect humans and, by ill-defined mechanisms, take up lifelong infections in their hosts. Here, we report that SFVs encode multiple miRNAs via a noncanonical mode of biogenesis. The primary SFV miRNA transcripts (pri-miRNAs) are transcribed by RNA polymerase III (RNAP III) and take multiple forms, including some that are cleaved by Drosha. However, these miRNAs are generated in a context-dependent fashion, as longer RNAP II transcripts spanning this region are resistant to Drosha cleavage. This suggests that the virus may avoid any fitness penalty that could be associated with viral genome/transcript cleavage. Two SFV miRNAs share sequence similarity and functionality with notable host miRNAs, the lymphoproliferative miRNA miR-155 and the innate immunity suppressor miR-132. These results have important implications regarding foamy virus biology, viral miRNAs, and the development of retroviral-based vectors. IMPORTANCE Fundamental questions remain about which viruses encode miRNAs and their associated functions. Currently, few natural viruses with RNA genomes have been reported to encode miRNAs. Simian foamy viruses are retroviruses that are prevalent in nonhuman host populations, and some can zoonotically infect humans who hunt primates or work as animal caretakers. We identify a cluster of miRNAs encoded by SFV. Characterization of these miRNAs reveals evolutionarily conserved, unconventional mechanisms to generate small RNAs. Several SFV miRNAs share sequence similarity and functionality with host miRNAs, including the oncogenic miRNA miR-155 and innate immunity suppressor miR-132. Strikingly, unrelated herpesviruses also tap into one or both of these same regulatory pathways, implying relevance to a broad range of viruses. These findings provide new insights with respect to foamy virus biology and vectorology. KW - MIR-155 KW - simian foamy viruses KW - long terminal repeat KW - viral microRNAs KW - RNA KW - infection KW - herpesvirus KW - recognition KW - prediction KW - ortholog Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-117216 SN - 2150-7511 VL - 5 IS - 2 ER - TY - JOUR A1 - Heydarian, Motaharehsadat A1 - Yang, Tao A1 - Schweinlin, Matthias A1 - Steinke, Maria A1 - Walles, Heike A1 - Rudel, Thomas A1 - Kozjak-Pavlovic, Vera T1 - Biomimetic human tissue model for long-term study of Neisseria gonorrhoeae infection JF - Frontiers in Microbiology N2 - Gonorrhea is the second most common sexually transmitted infection in the world and is caused by Gram-negative diplococcus Neisseria gonorrhoeae. Since N. gonorrhoeae is a human-specific pathogen, animal infection models are only of limited use. Therefore, a suitable in vitro cell culture model for studying the complete infection including adhesion, transmigration and transport to deeper tissue layers is required. In the present study, we generated three independent 3D tissue models based on porcine small intestinal submucosa (SIS) scaffold by co-culturing human dermal fibroblasts with human colorectal carcinoma, endometrial epithelial, and male uroepithelial cells. Functional analyses such as transepithelial electrical resistance (TEER) and FITC-dextran assay indicated the high barrier integrity of the created monolayer. The histological, immunohistochemical, and ultra-structural analyses showed that the 3D SIS scaffold-based models closely mimic the main characteristics of the site of gonococcal infection in human host including the epithelial monolayer, the underlying connective tissue, mucus production, tight junction, and microvilli formation. We infected the established 3D tissue models with different N. gonorrhoeae strains and derivatives presenting various phenotypes regarding adhesion and invasion. The results indicated that the disruption of tight junctions and increase in interleukin production in response to the infection is strain and cell type-dependent. In addition, the models supported bacterial survival and proved to be better suitable for studying infection over the course of several days in comparison to commonly used Transwell® models. This was primarily due to increased resilience of the SIS scaffold models to infection in terms of changes in permeability, cell destruction and bacterial transmigration. In summary, the SIS scaffold-based 3D tissue models of human mucosal tissues represent promising tools for investigating N. gonorrhoeae infections under close-to-natural conditions. KW - 3D tissue model KW - small intestinal submucosa scaffold KW - co-culture KW - infection KW - Neisseria gonorrhoeae Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-197912 SN - 1664-302X VL - 10 IS - 1740 ER - TY - JOUR A1 - Heidrich, Benjamin A1 - Wiegand, Steffen B. A1 - Buggisch, Peter A1 - Hinrichsen, Holger A1 - Link, Ralph A1 - Möller, Bernd A1 - Böker, Klaus H. W. A1 - Teuber, Gerlinde A1 - Klinker, Hartwig A1 - Zehnter, Elmar A1 - Naumann, Uwe A1 - Busch, Heiner W. A1 - Maasoumy, Benjamin A1 - Baum, Undine A1 - Hardtke, Svenja A1 - Manns, Michael P. A1 - Wedemeyer, Heiner A1 - Petersen, Jörg A1 - Cornberg, Markus T1 - Treatment of Naive Patients with Chronic Hepatitis C Genotypes 2 and 3 with Pegylated Interferon Alpha and Ribavirin in a Real World Setting: Relevance for the New Era of DAA JF - PLOS ONE N2 - Evidence based clinical guidelines are implemented to treat patients efficiently that include efficacy, tolerability but also health economic considerations. This is of particular relevance to the new direct acting antiviral agents that have revolutionized treatment of chronic hepatitis C. For hepatitis C genotypes 2/3 interferon free treatment is already available with sofosbuvir plus ribavirin. However, treatment with sofosbuvir-based regimens is 10-20 times more expensive compared to pegylated interferon alfa and ribavirin (PegIFN/RBV). It has to be discussed if PegIFN/RBV is still an option for easy to treat patients. We assessed the treatment of patients with chronic hepatitis C genotypes 2/3 with PegIFN/RBV in a real world setting according to the latest German guidelines. Overall, 1006 patients were recruited into a prospective patient registry with 959 having started treatment. The intention-to-treat analysis showed poor SVR (GT2 61%, GT3 47%) while patients with adherence had excellent SVR in the per protocol analysis (GT2 96%, GT3 90%). According to guidelines, 283 patients were candidates for shorter treatment duration, namely a treatment of 16 weeks (baseline HCV-RNA <800.000 IU/mL, no cirrhosis and RVR). However, 65% of these easy to treat patients have been treated longer than recommended that resulted in higher costs but not higher SVR rates. In conclusion, treatment with PegIFN/RBV in a real world setting can be highly effective yet similar effective than PegIFN +/- sofosbuvir/RBV in well-selected naive G2/3 patients. Full adherence to guidelines could be further improved, because it would be important in the new era with DAA, especially to safe resources. KW - peginterferon alpha-2B KW - HCV genotype-2 KW - sofosbuvir KW - infection KW - epidemology Y1 - 2014 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-115149 SN - 1932-6203 VL - 9 IS - 10 ER - TY - JOUR A1 - Heidrich, Benjamin A1 - Cordes, Hans-Jörg A1 - Klinker, Hartwig A1 - Möller, Bernd A1 - Naumann, Uwe A1 - Rössle, Martin A1 - Kraus, Michael R. A1 - Böker, Klaus H. A1 - Roggel, Christoph A1 - Schuchmann, Marcus A1 - Stoehr, Albrecht A1 - Trein, Andreas A1 - Hardtke, Svenja A1 - Gonnermann, Andrea A1 - Koch, Armin A1 - Wedemeyer, Heiner A1 - Manns, Michael P. A1 - Cornberg, Markus T1 - Treatment Extension of Pegylated Interferon Alpha and Ribavirin Does Not Improve SVR in Patients with Genotypes 2/3 without Rapid Virological Response (OPTEX Trial): A Prospective, Randomized, Two-Arm, Multicentre Phase IV Clinical Trial JF - PLoS ONE N2 - Although sofosbuvir has been approved for patients with genotypes 2/3 (G2/3), many parts of the world still consider pegylated Interferon alpha (P) and ribavirin (R) as standard of care for G2/3. Patients with rapid virological response (RVR) show response rates >80%. However, SVR (sustained virological response) in non-RVR patients is not satisfactory. Longer treatment duration may be required but evidence from prospective trials are lacking. A total of 1006 chronic HCV genotype 2/3 patients treated with P/R were recruited into a German HepNet multicenter screening registry. Of those, only 226 patients were still HCV RNA positive at week 4 (non-RVR). Non-RVR patients with ongoing response after 24 weeks P-2b/R qualified for OPTEX, a randomized trial investigating treatment extension of additional 24 weeks (total 48 weeks, Group A) or additional 12 weeks (total 36 weeks, group B) of 1.5 \(\mu\)g/kg P-2b and 800-1400 mg R. Due to the low number of patients without RVR, the number of 150 anticipated study patients was not met and only 99 non-RVR patients (n=50 Group A, n=49 Group B) could be enrolled into the OPTEX trial. Baseline factors did not differ between groups. Sixteen patients had G2 and 83 patients G3. Based on the ITT (intention-to-treat) analysis, 68% [55%; 81%] in Group A and 57% [43%; 71%] in Group B achieved SVR (p=0.31). The primary endpoint of better SVR rates in Group A compared to a historical control group (SVR 70%) was not met. In conclusion, approximately 23% of G2/3 patients did not achieve RVR in a real world setting. However, subsequent recruitment in a treatment-extension study was difficult. Prolonged therapy beyond 24 weeks did not result in higher SVR compared to a historical control group. KW - chronic hepatitis C KW - peginterferon alpha-2b KW - infection KW - sofosbuvir KW - therapy KW - HCV genotype 2 Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151811 VL - 10 IS - 6 ER - TY - JOUR A1 - Glaser, Kirsten A1 - Silwedel, Christine A1 - Fehrholz, Markus A1 - Waaga-Gasser, Ana M. A1 - Henrich, Birgit A1 - Claus, Heike A1 - Speer, Christian P. T1 - Ureaplasma Species Differentially Modulate Pro- and Anti-Inflammatory Cytokine Responses in Newborn and Adult Human Monocytes Pushing the State Toward Pro-Inflammation JF - Frontiers in Cellular and Infection Microbiology N2 - Background: Ureaplasma species have been associated with chorioamnionitis and preterm birth and have been implicated in the pathogenesis of neonatal short and long-term morbidity. However, being mostly commensal bacteria, controversy remains on the pro-inflammatory capacity of Ureaplasma. Discussions are ongoing on the incidence and impact of prenatal, perinatal, and postnatal infection. The present study addressed the impact of Ureaplasma isolates on monocyte-driven inflammation. Methods: Cord blood monocytes of term neonates and adult monocytes, either native or LPS-primed, were cultured with Ureaplasma urealyticum (U. urealyticum) serovar 8 (Uu8) and Ureaplasma parvum serovar 3 (Up3). Using qRT-PCR, cytokine flow cytometry, and multi-analyte immunoassay, we assessed mRNA and protein expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-8, IL-12p40, IL-10, and IL-1 receptor antagonist (IL-1ra) as well as Toll-like receptor (TLR) 2 and TLR4. Results: Uu8 and Up3 induced mRNA expression and protein release of TNF-α, IL-1β and IL-8 in term neonatal and adult monocytes (p < 0.01 and p < 0.05). Intracellular protein expression of TNF-α, IL-1β and IL-8 in Ureaplasma-stimulated cells paralleled those results. Ureaplasma-induced cytokine levels did not significantly differ from LPS-mediated levels except for lower intracellular IL-1β in adult monocytes (Uu8: p < 0.05). Remarkably, ureaplasmas did not induce IL-12p40 response and promoted lower amounts of anti-inflammatory IL-10 and IL-1ra than LPS, provoking a cytokine imbalance more in favor of pro-inflammation (IL-1β/IL-10, IL-8/IL-10 and IL-8/IL-1ra: p < 0.01, vs. LPS). In contrast to LPS, both isolates induced TLR2 mRNA in neonatal and adult cells (p < 0.001 and p < 0.05) and suppressed TLR4 mRNA in adult monocytes (p < 0.05). Upon co-stimulation, Uu8 and Up3 inhibited LPS-induced intracellular IL-1β (p < 0.001 and p < 0.05) and IL-8 in adult monocytes (p < 0.01), while LPS-induced neonatal cytokines were maintained or aggravated (p < 0.05). Conclusion: Our data demonstrate a considerable pro-inflammatory capacity of Ureaplasma isolates in human monocytes. Stimulating pro-inflammatory cytokine responses while hardly inducing immunomodulatory and anti-inflammatory cytokines, ureaplasmas might push monocyte immune responses toward pro-inflammation. Inhibition of LPS-induced cytokines in adult monocytes in contrast to sustained inflammation in term neonatal monocytes indicates a differential modulation of host immune responses to a second stimulus. Modification of TLR2 and TLR4 expression may shape host susceptibility to inflammation. KW - Ureaplasma KW - infection KW - inflammation KW - immunomodulation KW - chorioamnionitis KW - neonatal morbidity KW - monocytes KW - cord blood Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-169958 VL - 7 IS - 484 ER - TY - JOUR A1 - Gerova, Milan A1 - Wicke, Laura A1 - Chihara, Kotaro A1 - Schneider, Cornelius A1 - Lavigne, Rob A1 - Vogel, Jörg T1 - A grad-seq view of RNA and protein complexes in Pseudomonas aeruginosa under standard and bacteriophage predation conditions JF - mbio N2 - The Gram-negative rod-shaped bacterium Pseudomonas aeruginosa is not only a major cause of nosocomial infections but also serves as a model species of bacterial RNA biology. While its transcriptome architecture and posttranscriptional regulation through the RNA-binding proteins Hfq, RsmA, and RsmN have been studied in detail, global information about stable RNA-protein complexes in this human pathogen is currently lacking. Here, we implement gradient profiling by sequencing (Grad-seq) in exponentially growing P. aeruginosa cells to comprehensively predict RNA and protein complexes, based on glycerol gradient sedimentation profiles of >73% of all transcripts and ∼40% of all proteins. As to benchmarking, our global profiles readily reported complexes of stable RNAs of P. aeruginosa, including 6S RNA with RNA polymerase and associated product RNAs (pRNAs). We observe specific clusters of noncoding RNAs, which correlate with Hfq and RsmA/N, and provide a first hint that P. aeruginosa expresses a ProQ-like FinO domain-containing RNA-binding protein. To understand how biological stress may perturb cellular RNA/protein complexes, we performed Grad-seq after infection by the bacteriophage ΦKZ. This model phage, which has a well-defined transcription profile during host takeover, displayed efficient translational utilization of phage mRNAs and tRNAs, as evident from their increased cosedimentation with ribosomal subunits. Additionally, Grad-seq experimentally determines previously overlooked phage-encoded noncoding RNAs. Taken together, the Pseudomonas protein and RNA complex data provided here will pave the way to a better understanding of RNA-protein interactions during viral predation of the bacterial cell. IMPORTANCE Stable complexes by cellular proteins and RNA molecules lie at the heart of gene regulation and physiology in any bacterium of interest. It is therefore crucial to globally determine these complexes in order to identify and characterize new molecular players and regulation mechanisms. Pseudomonads harbor some of the largest genomes known in bacteria, encoding ∼5,500 different proteins. Here, we provide a first glimpse on which proteins and cellular transcripts form stable complexes in the human pathogen Pseudomonas aeruginosa. We additionally performed this analysis with bacteria subjected to the important and frequently encountered biological stress of a bacteriophage infection. We identified several molecules with established roles in a variety of cellular pathways, which were affected by the phage and can now be explored for their role during phage infection. Most importantly, we observed strong colocalization of phage transcripts and host ribosomes, indicating the existence of specialized translation mechanisms during phage infection. All data are publicly available in an interactive and easy to use browser. KW - Grad-seq KW - Pseudomonas KW - UKZ KW - bacteriophage KW - infection KW - Pseudomonas aeruginosa KW - RNA-binding proteins KW - noncoding RNA KW - phage Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259054 VL - 12 IS - 1 ER - TY - JOUR A1 - García-Betancur, Juan-Carlos A1 - Goñi-Moreno, Angel A1 - Horger, Thomas A1 - Schott, Melanie A1 - Sharan, Malvika A1 - Eikmeier, Julian A1 - Wohlmuth, Barbara A1 - Zernecke, Alma A1 - Ohlsen, Knut A1 - Kuttler, Christina A1 - Lopez, Daniel T1 - Cell differentiation defines acute and chronic infection cell types in Staphylococcus aureus JF - eLife N2 - A central question to biology is how pathogenic bacteria initiate acute or chronic infections. Here we describe a genetic program for cell-fate decision in the opportunistic human pathogen Staphylococcus aureus, which generates the phenotypic bifurcation of the cells into two genetically identical but different cell types during the course of an infection. Whereas one cell type promotes the formation of biofilms that contribute to chronic infections, the second type is planktonic and produces the toxins that contribute to acute bacteremia. We identified a bimodal switch in the agr quorum sensing system that antagonistically regulates the differentiation of these two physiologically distinct cell types. We found that extracellular signals affect the behavior of the agr bimodal switch and modify the size of the specialized subpopulations in specific colonization niches. For instance, magnesium-enriched colonization niches causes magnesium binding to S. aureusteichoic acids and increases bacterial cell wall rigidity. This signal triggers a genetic program that ultimately downregulates the agr bimodal switch. Colonization niches with different magnesium concentrations influence the bimodal system activity, which defines a distinct ratio between these subpopulations; this in turn leads to distinct infection outcomes in vitro and in an in vivo murine infection model. Cell differentiation generates physiological heterogeneity in clonal bacterial infections and helps to determine the distinct infection types. KW - Staphylococcus aureus KW - infection KW - cell differentiation KW - pathogenic bacteria Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170346 VL - 6 IS - e28023 ER - TY - JOUR A1 - Dühring, Sybille A1 - Germerodt, Sebastian A1 - Skerka, Christine A1 - Zipfel, Peter F. A1 - Dandekar, Thomas A1 - Schuster, Stefan T1 - Host-pathogen interactions between the human innate immune system and Candida albicans - understanding and modeling defense and evasion strategies JF - Frontiers in Microbiology N2 - The diploid, polymorphic yeast Candida albicans is one of the most important human pathogenic fungi. C. albicans can grow, proliferate and coexist as a commensal on or within the human host for a long time. However, alterations in the host environment can render C. albicans virulent. In this review, we describe the immunological cross-talk between C. albicans and the human innate immune system. We give an overview in form of pairs of human defense strategies including immunological mechanisms as well as general stressors such as nutrient limitation, pH, fever etc. and the corresponding fungal response and evasion mechanisms. Furthermore, Computational Systems Biology approaches to model and investigate these complex interactions are highlighted with a special focus on game-theoretical methods and agent-based models. An outlook on interesting questions to be tackled by Systems Biology regarding entangled defense and evasion mechanisms is given. KW - agent-based model KW - antimicrobial peptides KW - fungal pathogens KW - Candida albicans KW - immunological cross-talk KW - beta-lactamase inhibition KW - in vitro KW - biomaterial surfaces KW - biofilm formation KW - dendritic cells KW - infection KW - resistance KW - human immune system KW - host-pathogen interaction KW - computational systems biology KW - defense and evasion strategies Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-151621 VL - 6 IS - 625 ER - TY - JOUR A1 - Däullary, Thomas A1 - Imdahl, Fabian A1 - Dietrich, Oliver A1 - Hepp, Laura A1 - Krammer, Tobias A1 - Fey, Christina A1 - Neuhaus, Winfried A1 - Metzger, Marco A1 - Vogel, Jörg A1 - Westermann, Alexander J. A1 - Saliba, Antoine-Emmanuel A1 - Zdzieblo, Daniela T1 - A primary cell-based in vitro model of the human small intestine reveals host olfactomedin 4 induction in response to Salmonella Typhimurium infection JF - Gut Microbes N2 - Infection research largely relies on classical cell culture or mouse models. Despite having delivered invaluable insights into host-pathogen interactions, both have limitations in translating mechanistic principles to human pathologies. Alternatives can be derived from modern Tissue Engineering approaches, allowing the reconstruction of functional tissue models in vitro. Here, we combined a biological extracellular matrix with primary tissue-derived enteroids to establish an in vitro model of the human small intestinal epithelium exhibiting in vivo-like characteristics. Using the foodborne pathogen Salmonella enterica serovar Typhimurium, we demonstrated the applicability of our model to enteric infection research in the human context. Infection assays coupled to spatio-temporal readouts recapitulated the established key steps of epithelial infection by this pathogen in our model. Besides, we detected the upregulation of olfactomedin 4 in infected cells, a hitherto unrecognized aspect of the host response to Salmonella infection. Together, this primary human small intestinal tissue model fills the gap between simplistic cell culture and animal models of infection, and shall prove valuable in uncovering human-specific features of host-pathogen interplay. KW - intestinal enteroids KW - biological scaffold KW - Salmonella Typhimurium KW - OLFM4 KW - NOTCH KW - filamentous Salmonella Typhimurium KW - bacterial migration KW - bacterial virulence KW - 3D tissue model KW - olfactomedin 4 KW - infection Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-350451 VL - 15 IS - 1 ER - TY - JOUR A1 - Dembek, Marcin A1 - Barquist, Lars A1 - Boinett, Christine J. A1 - Cain, Amy K. A1 - Mayho, Matthew A1 - Lawley, Trevor D. A1 - Fairweather, Neil F. A1 - Fagan, Robert P. T1 - High-throughput analysis of gene essentiality and sporulation in Clostridium difficile JF - mBio N2 - Clostridium difficile is the most common cause of antibiotic-associated intestinal infections and a significant cause of morbidity and mortality. Infection with C. difficile requires disruption of the intestinal microbiota, most commonly by antibiotic usage. Therapeutic intervention largely relies on a small number of broad-spectrum antibiotics, which further exacerbate intestinal dysbiosis and leave the patient acutely sensitive to reinfection. Development of novel targeted therapeutic interventions will require a detailed knowledge of essential cellular processes, which represent attractive targets, and species-specific processes, such as bacterial sporulation. Our knowledge of the genetic basis of C. difficile infection has been hampered by a lack of genetic tools, although recent developments have made some headway in addressing this limitation. Here we describe the development of a method for rapidly generating large numbers of transposon mutants in clinically important strains of C. difficile. We validated our transposon mutagenesis approach in a model strain of C. difficile and then generated a comprehensive transposon library in the highly virulent epidemic strain R20291 (027/BI/NAP1) containing more than 70,000 unique mutants. Using transposon-directed insertion site sequencing (TraDIS), we have identified a core set of 404 essential genes, required for growth in vitro. We then applied this technique to the process of sporulation, an absolute requirement for C. difficile transmission and pathogenesis, identifying 798 genes that are likely to impact spore production. The data generated in this study will form a valuable resource for the community and inform future research on this important human pathogen. KW - Bacillus subtilis KW - expression KW - spores KW - toxin KW - transcription KW - germination KW - transposition KW - metabolism KW - infection KW - in vitro Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143745 VL - 6 IS - 2 ER - TY - JOUR A1 - Dasenbrook, Elliot C. A1 - Lu, Luan A1 - Donnola, Shannon A1 - Weaver, David E. A1 - Gulani, Viskas A1 - Jakob, Peter M. A1 - Konstan, Michael W. A1 - Flask, Chris A. T1 - Normalized T1 Magnetic Resonance Imaging for Assessment of Regional Lung Function in Adult Cystic Fibrosis Patients - A Cross-Sectional Study JF - PLOS ONE N2 - Background: Cystic fibrosis (CF) patients would benefit from a safe and effective tool to detect early-stage, regional lung disease to allow for early intervention. Magnetic Resonance Imaging (MRI) is a safe, non-invasive procedure capable of providing quantitative assessments of disease without ionizing radiation. We developed a rapid normalized T1 MRI technique to detect regional lung disease in early-stage CF patients. Materials and Methods: Conventional multislice, pulmonary T1 relaxation time maps were obtained for 10 adult CF patients with normal spirometry and 5 healthy non-CF control subjects using a rapid Look-Locker MRI acquisition (5 seconds/imaging slice). Each lung absolute T1 map was separated into six regions of interest (ROI) by manually selecting upper, central, and lower lung regions in the left and right lungs. In order to reduce the effects of subject-to-subject variation, normalized T1 maps were calculated by dividing each pixel in the absolute T1 maps by the mean T1 time in the central lung region. The primary outcome was the differences in mean normalized T1 values in the upper lung regions between CF patients with normal spirometry and healthy volunteers. Results: Normalized T1 (nT1) maps showed visibly reduced subject-to-subject variation in comparison to conventional absolute T1 maps for healthy volunteers. An ROI analysis showed that the variation in the nT1 values in all regions was <= 2% of the mean. The primary outcome, the mean (SD) of the normalized T1 values in the upper right lung regions, was significantly lower in the CF subjects [.914 (.037)] compared to the upper right lung regions of the healthy subjects [.983 (.003)] [difference of .069 (95% confidence interval .032-.105); p=.001). Similar results were seen in the upper left lung region. Conclusion: Rapid normalized T1 MRI relaxometry obtained in 5 seconds/imaging slice may be used to detect regional early-stage lung disease in CF patients. KW - infants KW - disease KW - ventilation KW - infection KW - guidelines KW - diagnosis KW - children Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-128346 SN - 1932-6203 VL - 8 IS - 9 ER - TY - JOUR A1 - Correia Santos, Sara A1 - Bischler, Thorsten A1 - Westermann, Alexander J. A1 - Vogel, Jörg T1 - MAPS integrates regulation of actin-targeting effector SteC into the virulence control network of Salmonella small RNA PinT JF - Cell Reports N2 - A full understanding of the contribution of small RNAs (sRNAs) to bacterial virulence demands knowledge of their target suites under infection-relevant conditions. Here, we take an integrative approach to capturing targets of the Hfq-associated sRNA PinT, a known post-transcriptional timer of the two major virulence programs of Salmonella enterica. Using MS2 affinity purification and RNA sequencing (MAPS), we identify PinT ligands in bacteria under in vitro conditions mimicking specific stages of the infection cycle and in bacteria growing inside macrophages. This reveals PinT-mediated translational inhibition of the secreted effector kinase SteC, which had gone unnoticed in previous target searches. Using genetic, biochemical, and microscopic assays, we provide evidence for PinT-mediated repression of steC mRNA, eventually delaying actin rearrangements in infected host cells. Our findings support the role of PinT as a central post-transcriptional regulator in Salmonella virulence and illustrate the need for complementary methods to reveal the full target suites of sRNAs. KW - gene expression KW - nondocing RNA KW - chaperone HFQ KW - soluble-RNA KW - SEQ KW - interactome KW - repression KW - secretion KW - infection KW - biology Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259134 VL - 34 IS - 5 ER - TY - JOUR A1 - Chumduri, Cindrilla A1 - Turco, Margherita Y. T1 - Organoids of the female reproductive tract JF - Journal of Molecular Medicine N2 - Healthy functioning of the female reproductive tract (FRT) depends on balanced and dynamic regulation by hormones during the menstrual cycle, pregnancy and childbirth. The mucosal epithelial lining of different regions of the FRT—ovaries, fallopian tubes, uterus, cervix and vagina—facilitates the selective transport of gametes and successful transfer of the zygote to the uterus where it implants and pregnancy takes place. It also prevents pathogen entry. Recent developments in three-dimensional (3D) organoid systems from the FRT now provide crucial experimental models that recapitulate the cellular heterogeneity and physiological, anatomical and functional properties of the organ in vitro. In this review, we summarise the state of the art on organoids generated from different regions of the FRT. We discuss the potential applications of these powerful in vitro models to study normal physiology, fertility, infections, diseases, drug discovery and personalised medicine. KW - female reproductive tract KW - organoids KW - reproductive health KW - pregnancy KW - fertility KW - infection KW - cancers Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-328374 VL - 99 IS - 4 ER - TY - JOUR A1 - Bakari-Soale, Majeed A1 - Ikenga, Nonso Josephat A1 - Scheibe, Marion A1 - Butter, Falk A1 - Jones, Nicola G. A1 - Kramer, Susanne A1 - Engstler, Markus T1 - The nucleolar DExD/H protein Hel66 is involved in ribosome biogenesis in Trypanosoma brucei JF - Scientific Reports N2 - The biosynthesis of ribosomes is a complex cellular process involving ribosomal RNA, ribosomal proteins and several further trans-acting factors. DExD/H box proteins constitute the largest family of trans-acting protein factors involved in this process. Several members of this protein family have been directly implicated in ribosome biogenesis in yeast. In trypanosomes, ribosome biogenesis differs in several features from the process described in yeast. Here, we have identified the DExD/H box helicase Hel66 as being involved in ribosome biogenesis. The protein is unique to Kinetoplastida, localises to the nucleolus and its depletion via RNAi caused a severe growth defect. Loss of the protein resulted in a decrease of global translation and accumulation of rRNA processing intermediates for both the small and large ribosomal subunits. Only a few factors involved in trypanosome rRNA biogenesis have been described so far and our findings contribute to gaining a more comprehensive picture of this essential process. KW - infection KW - parasite evolution KW - parasite genetics KW - RNA Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-263872 VL - 11 IS - 1 ER - TY - JOUR A1 - Ascierto, Maria Libera A1 - Worschech, Andrea A1 - Yu, Zhiya A1 - Adams, Sharon A1 - Reinboth, Jennifer A1 - Chen, Nanhai G A1 - Pos, Zoltan A1 - Roychoudhuri, Rahul A1 - Di Pasquale, Giovanni A1 - Bedognetti, Davide A1 - Uccellini, Lorenzo A1 - Rossano, Fabio A1 - Ascierto, Paolo A A1 - Stroncek, David F A1 - Restifo, Nicholas P A1 - Wang, Ena A1 - Szalay, Aladar A A1 - Marincola, Francesco M T1 - Permissivity of the NCI-60 cancer cell lines to oncolytic Vaccinia Virus GLV-1h68 JF - BMC Cancer N2 - Background: Oncolytic viral therapy represents an alternative therapeutic strategy for the treatment of cancer. We previously described GLV-1h68, a modified Vaccinia Virus with exclusive tropism for tumor cells, and we observed a cell line-specific relationship between the ability of GLV-1h68 to replicate in vitro and its ability to colonize and eliminate tumor in vivo. Methods: In the current study we surveyed the in vitro permissivity to GLV-1h68 replication of the NCI-60 panel of cell lines. Selected cell lines were also tested for permissivity to another Vaccinia Virus and a vesicular stomatitis virus (VSV) strain. In order to identify correlates of permissity to viral infection, we measured transcriptional profiles of the cell lines prior infection. Results: We observed highly heterogeneous permissivity to VACV infection amongst the cell lines. The heterogeneity of permissivity was independent of tissue with the exception of B cell derivation. Cell lines were also tested for permissivity to another Vaccinia Virus and a vesicular stomatitis virus (VSV) strain and a significant correlation was found suggesting a common permissive phenotype. While no clear transcriptional pattern could be identified as predictor of permissivity to infection, some associations were observed suggesting multifactorial basis permissivity to viral infection. Conclusions: Our findings have implications for the design of oncolytic therapies for cancer and offer insights into the nature of permissivity of tumor cells to viral infection. KW - gene-therapy KW - adenovirus KW - receptor KW - identification KW - infection KW - CD9 KW - panel Y1 - 2011 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-141503 VL - 11 IS - 451 ER - TY - JOUR A1 - Apsemidou, Athanasia A1 - Füller, Miriam Antonie A1 - Idelevich, Evgeny A. A1 - Kurzai, Oliver A1 - Tragiannidis, Athanasios A1 - Groll, Andreas H. T1 - Candida lusitaniae breakthrough fungemia in an immuno-compromised adolescent: case report and review of the literature JF - Journal of Fungi N2 - Candida lusitaniae is a rare cause of candidemia that is known for its unique capability to rapidly acquire resistance to amphotericin B. We report the case of an adolescent with grade IV graft-vs.-host disease after hematopoietic cell transplantation who developed catheter-associated C. lusitaniae candidemia while on therapeutic doses of liposomal amphotericin B. We review the epidemiology of C. lusitaniae bloodstream infections in adult and pediatric patients, the development of resistance, and its role in breakthrough candidemia. Appropriate species identification, in vitro susceptibility testing, and source control are pivotal to optimal management of C. lusitaniae candidemia. Initial antifungal therapy may consist of an echinocandin and be guided by in vitro susceptibility and clinical response. KW - Candida lusitaniae KW - candidemia KW - resistance KW - breakthrough KW - infection KW - transplantation Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-220125 SN - 2309-608X VL - 6 IS - 4 ER - TY - JOUR A1 - Afonso-Grunz, Fabian A1 - Hoffmeier, Klaus A1 - Müller, Sören A1 - Westermann, Alexander J. A1 - Rotter, Björn A1 - Vogel, Jörg A1 - Winter, Peter A1 - Kahl, Günter T1 - Dual 3'Seq using deepSuperSAGE uncovers transcriptomes of interacting Salmonella enterica Typhimurium and human host cells JF - BMC Genomics N2 - Background: The interaction of eukaryotic host and prokaryotic pathogen cells is linked to specific changes in the cellular proteome, and consequently to infection-related gene expression patterns of the involved cells. To simultaneously assess the transcriptomes of both organisms during their interaction we developed dual 3'Seq, a tag-based sequencing protocol that allows for exact quantification of differentially expressed transcripts in interacting pro-and eukaryotic cells without prior fixation or physical disruption of the interaction. Results: Human epithelial cells were infected with Salmonella enterica Typhimurium as a model system for invasion of the intestinal epithelium, and the transcriptional response of the infected host cells together with the differential expression of invading and intracellular pathogen cells was determined by dual 3'Seq coupled with the next-generation sequencing-based transcriptome profiling technique deepSuperSAGE (deep Serial Analysis of Gene Expression). Annotation to reference transcriptomes comprising the operon structure of the employed S. enterica Typhimurium strain allowed for in silico separation of the interacting cells including quantification of polycistronic RNAs. Eighty-nine percent of the known loci are found to be transcribed in prokaryotic cells prior or subsequent to infection of the host, while 75% of all protein-coding loci are represented in the polyadenylated transcriptomes of human host cells. Conclusions: Dual 3'Seq was alternatively coupled to MACE (Massive Analysis of cDNA ends) to assess the advantages and drawbacks of a library preparation procedure that allows for sequencing of longer fragments. Additionally, the identified expression patterns of both organisms were validated by qRT-PCR using three independent biological replicates, which confirmed that RELB along with NFKB1 and NFKB2 are involved in the initial immune response of epithelial cells after infection with S. enterica Typhimurium. KW - complete genome sequence KW - secretion systems KW - RNA-Seq KW - deepSuperSAGE KW - transcriptome KW - gene expression KW - serovar Typhimurium KW - human macrophages KW - epithelial cells KW - infection KW - SuperSAGE KW - receptors KW - Dual 3'seq KW - MACE KW - tag based KW - simultaneous KW - genome wide KW - gene expression profiling KW - host pathogen interaction KW - Salmonella enterica Typhimurium strain SL1344 Y1 - 2015 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143230 VL - 16 IS - 323 ER -