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The optimal treatment strategy for secondary prevention in patients with cryptogenic stroke and patent foramen ovale (PFO) has been a matter of controversy for decades. After three randomized trials failed to show a benefit of closure with an excess of complications in the interventional arm, two large recent trials suggest a benefit with regard of preventing further ischemic strokes. With this discrepancy in results it is important to discuss recent trials in detail and evolve an informed clinical approach for daily practice.
This review analyses the aesthetic engagement with Nazi atrocities during WWII and belonging in post-war Germany as presented in Nora Krug’s graphic novel Heimat: A German Family Album. The authors employ Marianne Hirsch’s concept of ‘postmemory’ as an analytical tool that helps them locate the complex historical and emotional contexts from which this graphic novel receives its impulses. The concrete scenes from the novel are presented and subsequently related to the field of memory and postmemory scholarship. Wider critical debates on how aesthetic articulations of past atrocities influence the next generations of ‘victims’ and ‘perpetrators’ are examined, to ask: What does it mean to inhabit memories of ghostly narratives about perpetrators and how does it form a feeling of post-home?
This dissertation is an investigation of pitch accent, or lexical tone, in standard Croatian. The first chapter presents an in-depth overview of the history of the Croatian language, its relationship to Serbo-Croatian, its dialect groups and pronunciation variants, and general phonology. The second chapter explains the difference between various types of prosodic prominence and describes systems of pitch accent in various languages from different parts of the world: Yucatec Maya, Lithuanian and Limburgian. Following is a detailed account of the history of tone in Serbo-Croatian and Croatian, the specifics of its tonal system, intonational phonology and finally, a review of the most prominent phonetic investigations of tone in that language.
The focal point of this dissertation is a production experiment, in which ten native speakers of Croatian from the region of Slavonia were recorded. The material recorded included a diverse selection of monosyllabic, bisyllabic, trisyllabic and quadrisyllabic words, containing all four accents of standard Croatian: short falling, long falling, short rising and long rising. Each target word was spoken in initial, medial and final positions of natural Croatian sentences. This research fills several gaps in the existing literature. Namely, the production of tone was investigated in words with a syllabic /r̩/, in pretonal syllables and in non-initial context. Acoustic parameters measured included duration, F0 in every 10% of the nucleus duration, overall pitch, pitch range and pitch peak alignment.
Results showed that differences between falling and rising accents in Croatian are produced mainly with tonal parameters and that the most salient features were pitch peak alignment and overall pitch. The difference between long and short accents was primarily durational and optionally tonal. Words produced in initial and medial sentence positions had a rising contour in their accented syllable, while in the final, segments were usually falling.
Progression of pupil dilation (PD) in response to visual stimuli may indicate distinct internal processes. No study has been performed on PD progression during a social cognition task. Here, we describe PD progression during the Movie for the Assessment of Social Cognition (MASC) test in n = 23 adolescents with Autism Spectrum Disorder (ASD) and n = 24 age, IQ and sex‐matched neurotypical controls (NTC). The MASC consists of 43 video sequences depicting human social interactions, each followed by a multiple‐choice question concerning characters' mental states. PD progression data were extracted by eye tracking and controlled for fixation behavior. Segmenting PD progression during video sequences by principal component analysis, three sequential PD components were unveiled. In ASD compared with NTC, a distinct PD progression was observed with increased constriction amplitude, increased dilation latency, and increased dilation amplitude that correlated with PD progression components. These components predicted social cognition performance. The first and second PD components correlated positively with MASC behavioral performance in ASD but negatively in NTC. These PD components may be interpreted as indicators of sensory‐perceptual processing and attention function. In ASD, aberrant sensory‐perceptual processing and attention function could contribute to attenuated social cognition performance. This needs to be tested by additional studies combining the respective cognitive tests and the outlined PD progression analysis. Phasic activity of the locus coeruleus–norepinephrine system is discussed as putatively shared underlying mechanism.
Background. Extracts from Viscum album L. (VE) are used in the complementary cancer therapy in Europe for decades. VE contain several compounds like the mistletoe lectins (MLs) 1-3 and viscotoxins and also several minor ingredients. Since mistletoe lectin 1 (ML-1) has been described as the main component of VE harboring antitumor activity, purified native or recombinant ML-1 has been recently used in clinical trials. MLs stimulate the immune system, induce cytotoxicity, are able to modify the expression of cancer-associated genes, and influence the proliferation and motility of tumor cells.
Objective. In this study our goal was to determine anticancer effects of the VE ISCADOR Qu, of recombinant ML-1 (Aviscumine), and of native ML-1 in the treatment of glioblastoma (GBM), the most common and highly malignant brain tumor in adults. Additionally we were interested whether these drugs, used in combination with a temozolomide-(TMZ)-based radio-chemotherapy, provide synergistic effects.
Methods. Cell culture assays, ex vivo murine hippocampal brain slice cultures, human GBM cryosections, and a xenograft orthotopic glioblastoma mouse model were used.
Results. In cells, the expression of the ML receptor CD75s, which is also expressed in GBM specimen, but not in normal brain, correlates with the drug-induced cytotoxicity. In GBM cells, the drugs induce cell death in a concentration-dependent manner and reduce cell growth by inducing cell cycle arrest in the G2/M phase. The cell cycle arrest was paralleled by modifications in the expression of cell cycle regulating genes. ML containing drugs, if combined with glioma standard therapy, provide synergistic and additive anticancer effects. Despite not reaching statistical significance, a single intratumoral application of Aviscumine prolonged the median survival of GBM mice longer than tumor irradiation. Moreover, intratumorally applied Aviscumine prolonged the survival of GBM-bearing mice if used in combination with irradiation and TMZ for further 6.5 days compared to the radio-chemotherapy.
Conclusion. Our results suggest that an adjuvant treatment of glioma patients with ML-containing drugs might be beneficial.
Background: Despite significant advances in the understanding of glioblastoma genetics and biology, survival is still poor. Hypoxia and nutrient depletion in the tumour microenvironment induce adaptive signalling and metabolic responses, which can influence sensitivity to therapeutic regimens. DNA damage-inducible transcript 4 (DDIT4) is a protein induced by hypoxia and in response to DNA stress. Mechanistically, DDIT4 inhibits mammalian target of rapamycin complex 1 (mTORC1) signalling by activation of the tuberous sclerosis 1/2 (TSC1/2) complex.
Methods: Using short hairpin RNA-mediated gene suppression as well as doxycycline-regulated gene induction, we developed a glioblastoma cell model to study effects of DDIT4 under conditions of the glioblastoma microenvironment and therapy.
Results: We found an intact DDIT4-mTORC1 signalling axis in human glioblastoma cells that was inducible by hypoxia. Temozolomide and radiotherapy also induced DDIT4 and repressed mTORC1 activity in some glioblastoma cell lines. DDIT4 gene suppression sensitised glioma cells towards hypoxia-induced cell death, while DDIT4 overexpression protected them. Additionally, in clonogenic survival analyses, DDIT4 induction conferred protection from radiotherapy and temozolomide, while DDIT4 gene suppression sensitised cells.
Conclusions: We identified DDIT4 as a cell-intrinsic regulator for adaptive responses and therapy resistance in glioblastoma cells which may interfere with cell death induction by temozolomide, radiotherapy or hypoxia by inhibiting mTORC1 activity.
KDEL receptors (KDELRs) represent transmembrane proteins of the secretory pathway which regulate the retention of soluble ER-residents as well as retrograde and anterograde vesicle trafficking. In addition, KDELRs are involved in the regulation of cellular stress response and ECM degradation. For a deeper insight into KDELR1 specific functions, we characterised a KDELR1-KO cell line (HAP1) through whole transcriptome analysis by comparing KDELR1-KO cells with its respective HAP1 wild-type. Our data indicate more than 300 significantly and differentially expressed genes whose gene products are mainly involved in developmental processes such as cell adhesion and ECM composition, pointing out to severe cellular disorders due to a loss of KDELR1. Impaired adhesion capacity of KDELR1-KO cells was further demonstrated through in vitro adhesion assays, while collagen- and/or laminin-coating nearly doubled the adhesion property of KDELR1-KO cells compared to wild-type, confirming a transcriptional adaptation to improve or restore the cellular adhesion capability. Perturbations within the secretory pathway were verified by an increased secretion of ER-resident PDI and decreased cell viability under ER stress conditions, suggesting KDELR1-KO cells to be severely impaired in maintaining cellular homeostasis.
Wetlands such as bogs, swamps, or freshwater marshes are hotspots of biodiversity. For 5.1 million km2 of inland wetlands, the dynamics of area and water storage, which strongly impact biodiversity and ecosystem services, were simulated using the global hydrological model WaterGAP. For the first time, the impacts of both human water use and man‐made reservoirs (WUR) and future climate change (CC) on wetlands around the globe were quantified. WUR impacts are concentrated in arid/semiarid regions, where WUR decreased mean wetland water storage by more than 5% on 8.2% of the mean wetland area during 1986–2005 (Am), with highest decreases in groundwater depletion area. Using output of three climate models, CC impacts on wetlands were quantified, distinguishing unavoidable impacts [i.e., at 2 °C global warming (GW)] from avoidable impacts (difference between 3 °C and 2 °C impacts). Even unavoidable CC impacts are projected to be much larger than WUR impacts, also in arid/semiarid regions. On most wetland area with reliable estimates, avoidable CC impacts are more than twice as large as unavoidable impacts. In case of 2 °C GW, half of Am is estimated to be unaffected by mean storage changes of more than 5%, but only one third in case of 3 °C GW. Temporal variability of water storage will increase for most wetlands. Wetlands in dry regions will be affected the most, particularly by water storage decreases in the dry season. Different from wealthier countries, low‐income countries will dominantly suffer from a decrease in wetland water storage due to CC.
The endosteal bone marrow niche and vascular endothelial cells provide sanctuaries to leukemic cells. In murine chronic myeloid leukemia (CML) CD44 on leukemia cells and E-selectin on bone marrow endothelium are essential mediators for the engraftment of leukemic stem cells (LSC). We hypothesized that non-adhesion of CML-initiating cells to E-selectin on the bone marrow endothelium may lead to superior eradication of LSC in CML after treatment with imatinib than imatinib alone. Indeed, here we show that treatment with the E-selectin inhibitor GMI-1271 in combination with imatinib prolongs survival of mice with CML via decreased contact time of leukemia cells with bone marrow endothelium. Non-adhesion of BCR-ABL1+ cells leads to an increase of cell cycle progression and an increase of expression of the hematopoietic transcription factor and protooncogene Scl/Tal1 in leukemia-initiating cells (LIC). We implicate SCL/TAL1 as indirect phosphorylation target of BCR-ABL1 and as a negative transcriptional regulator of CD44 expression. We show that increased SCL/TAL1 expression is associated with improved outcome in human CML. These data demonstrate the BCR-ABL1-specific, cell-intrinsic pathways leading to altered interactions with the vascular niche via the modulation of adhesion molecules - a strategy therapeutically exploitable in future.
Die Plasmamembran eukaryotischer Zellen dient als Barriere zwischen dem Inneren einer Zelle und ihrer Umgebung. Eine wichtige Aufgabe von Proteinen, die sich in der Plasmamembran befinden, besteht in der Erkennung der Umgebung, der Übermittlung dieser Informationen über die Plasmamembran in das Innere einer Zelle und der Einleitung einer zellulären Antwort. Membranrezeptoren binden Liganden, was zu ihrer Aktivierung und der Rekrutierung von intrazellulären Proteinen führt. Funktionelle Signalkomplexe werden gebildet und leiten einen Informationstransfer durch die Zellmembran ein, so dass die Expression bestimmter Gene stimuliert oder unterdrückt wird. Eine Störung der Signalinitiierung und -übertragung tritt bei vielen Krankheiten auf, so dass Membranproteine ein wichtiges Ziel in der Medikamentenentwicklung sind.
In dieser Arbeit wird die Fragestellung bearbeitet, wie der Tumornekrosefaktor-Rezeptor 1 (TNFR1) in funktionelle Komplexe in der Plasmamembran einer intakten Zelle organisiert ist. TNFR1 besitzt vier cysteinreiche Domänen (CRDs) in seiner extrazellulären Region. Die erste und von der Plasmamembran am weitesten entfernte CRD ist die Pre-Ligand Assembly Domain (PLAD). Kristallstrukturen zeigten, dass sich in einem TNFR1-Dimer zwei PLAD in unmittelbarer Nähe befinden. Crosslinking-Experimente berichteten über mehrere oligomere Zustände von TNFR1; die Ergebnisse unterschieden sich nach Art und Konzentration des Crosslinkers. In der nativen Umgebung einer intakten Zelle wurde der oligomere Zustand von TNFR1 bisher nicht bestimmt. Der kanonische Ligand für TNFR1 ist der Tumornekrosefaktor alpha (TNF), ein Homotrimer, welches in löslicher oder membrangebundener Form vorliegt. Nach der Bindung von TNF an TNFR1 bilden sich Rezeptortrimere. Diese Proteinkomplexe rekrutieren intrazellulär Proteine und bilden einen funktionellen Membrankomplex, der intrazelluläre Signalkaskaden aktiviert. Die kanonische Signalweiterleitung erfolgt durch den nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-B), welcher Zellteilung oder Entzündung induziert. TNFR1 kann auch andere Signalwege wie beispielsweise Apoptose durch einen zytosolischen Komplex und die Procaspase-8, oder Nekroptose durch das Nekrosom und die mixed lineage kinase domain-like (MLKL)-Domäne einleiten. Die Dysregulation von TNFR1 ist bei einer Vielzahl von Krankheiten zu finden. Erhöhte TNFR1-Expressiosraten treten bei acquired immune deficiency syndrome (AIDS), multipler Sklerose und verschiedenen Krebsarten auf.
In einem zweiten Projekt wurde in Zusammenarbeit mit Prof. Dr. Michael Lanzer (Heidelberg, Germany) der Expressionsgrad des Proteins VAR2CSA in membranassoziierten knobs bestimmt, welche in Erythrozyten vorkommen, die mit dem Parasiten Plasmodium falciparum infizierten wurden. VAR2CSA gehört zur Proteinfamilie des Plasmodium falciparum erythrocyte membrane protein 1 (pfEMP1). Nach einer Infektion wird VAR2CSA zur Wirtszellmembran transportiert und in knobs eingelagert. Patienten, die Sichelzellenanämie-Erythrozyten (HbAS) aufweisen, sind im Gegensatz zu Patienten mit gesunden Erythrozyten (HbAA) immun gegen Malaria. Während die beiden Erythrozytentypen eine unterschiedliche Morphologie der knobs aufweisen, blieb ihre Zusammensetzung in Bezug auf VAR2CSA bisher ungeklärt.
Das Verständnis der Proteinfunktion erfordert eine Beschreibung der molekularen Organisation funktioneller Einheiten in der zellulären Umgebung. Hierfür ist die Fluoreszenzmikroskopie eine geeignete Methode, da sie eine gezielte Markierung von Zielproteinen ermöglicht. Die hohe Sensitivität ermöglicht die Visualisierung einzelner Proteine. Eine Einschränkung in der konventionellen Fluoreszenzmikroskopie ist die Auflösungsgrenze. Strukturelle Elemente, die kleiner als etwa die halbe Anregungswellenlänge sind (für die meisten Anwendungen 200 bis 300 nm) können nicht aufgelöst werden. Die Entwicklung der hochauflösenden Fluoreszenzmikroskopie ermöglichte es, diese Auflösungsgrenze zu umgehen und eine räumliche Auflösung von wenigen Nanometern zu erreichen, was die Visualisierung und Charakterisierung einzelner Proteinkomplexe ermöglichte. Eine Art der hochauflösenden Fluoreszenzmikroskopie ist die single-molecule localization microscopy (SMLM), die auf der Detektion einzelner Fluorophore, einer genauen Bestimmung ihrer Position (Lokalisation) und der Erzeugung eines rekonstruierten Bildes unterhalb der optischen Auflösungsgrenze basiert. Da die meisten Proben in der Fluoreszenzmikroskopie eine zu hohe räumliche Dichte an Fluorophoren aufweisen, um den Nachweis von einzelnen Fluorophoren zu ermöglichen, werden Verfahren zur Kontrolle der Emission von Fluorophoren eingesetzt. Eine Möglichkeit ist der Einsatz von Fluorophoren, die optisch zwischen einem nicht-fluoreszierenden und einem fluoreszierenden Zustand geschaltet werden können, z.B. photoschaltbare fluoreszierende Proteine in photoactivated localization microscopy (PALM) oder organische Farbstoffe in (direct) stochastic optical reconstruction microscopy ((d)STORM). SMLM erreicht eine räumliche Auflösung von 20 nm, was in den meisten Fällen ausreicht, um einzelne Proteinkomplexe in einer Zelle aufzulösen. Diese räumliche Auflösung ist jedoch nicht ausreichend, um Untereinheiten innerhalb eines Proteinkomplexes zu visualisieren. Zu diesem Zweck wurde SMLM erweitert und die verfügbare kinetische Information genutzt, die bei der Detektion einzelner Fluorophore ausgelesen wird. Viele Fluorophore weisen metastabile Dunkelzustände auf, die eine Lebensdauer von bis zu Sekunden aufweisen. Diese Übergänge erscheinen als "Blinken" der Fluoreszenzemission. In Kombination mit kinetischen Modellen kann aus der Anzahl an Blink-Ereignissen die Anzahl der Fluorophore ermittelt werden. Angewendet auf hochaufgelöste Proteinkomplexe kann die Auflösungsgrenze von hochauflösender Mikroskopie umgangen werden, und die Anzahl der Protein-Untereinheiten in einem hochaufgelösten Proteincluster ermittelt werden. Hierzu wird beispielsweise das photoschaltbare fluoreszierende Protein mEos2 an ein Zielprotein funsioniert (quantitative PALM (qPALM)).
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Cancer-associated fibroblasts (CAFs) in the tumor microenvironment contribute to all stages of tumorigenesis and are usually considered to be tumor-promoting cells. CAFs show a remarkable degree of heterogeneity, which is attributed to developmental origin or to local environmental niches, resulting in distinct CAF subsets within individual tumors. While CAF heterogeneity is frequently investigated in late-stage tumors, data on longitudinal CAF development in tumors are lacking. To this end, we used the transgenic polyoma middle T oncogene-induced mouse mammary carcinoma model and performed whole transcriptome analysis in FACS-sorted fibroblasts from early- and late-stage tumors. We observed a shift in fibroblast populations over time towards a subset previously shown to negatively correlate with patient survival, which was confirmed by multispectral immunofluorescence analysis. Moreover, we identified a transcriptomic signature distinguishing CAFs from early- and late-stage tumors. Importantly, the signature of early-stage CAFs correlated well with tumor stage and survival in human mammary carcinoma patients. A random forest analysis suggested predictive value of the complete set of differentially expressed genes between early- and late-stage CAFs on bulk tumor patient samples, supporting the clinical relevance of our findings. In conclusion, our data show transcriptome alterations in CAFs during tumorigenesis in the mammary gland, which suggest that CAFs are educated by the tumor over time to promote tumor development. Moreover, we show that murine CAF gene signatures can harbor predictive value for human cancer.
uORF-tools—workflow for the determination of translation-regulatory upstream open reading frames
(2019)
Ribosome profiling (ribo-seq) provides a means to analyze active translation by determining ribosome occupancy in a transcriptome-wide manner. The vast majority of ribosome protected fragments (RPFs) resides within the protein-coding sequence of mRNAs. However, commonly reads are also found within the transcript leader sequence (TLS) (aka 5’ untranslated region) preceding the main open reading frame (ORF), indicating the translation of regulatory upstream ORFs (uORFs). Here, we present a workflow for the identification of translation-regulatory uORFs. Specifically, uORF-Tools uses Ribo-TISH to identify uORFs within a given dataset and generates a uORF annotation file. In addition, a comprehensive human uORF annotation file, based on 35 ribo-seq files, is provided, which can serve as an alternative input file for the workflow. To assess the translation-regulatory activity of the uORFs, stimulus-induced changes in the ratio of the RPFs residing in the main ORFs relative to those found in the associated uORFs are determined. The resulting output file allows for the easy identification of candidate uORFs, which have translation-inhibitory effects on their associated main ORFs. uORF-Tools is available as a free and open Snakemake workflow at https://github.com/Biochemistry1-FFM/uORF-Tools. It is easily installed and all necessary tools are provided in a version-controlled manner, which also ensures lasting usability. uORF-Tools is designed for intuitive use and requires only limited computing times and resources.
The interdependence of selective cues during development of regulatory T cells (Treg cells) in the thymus and their suppressive function remains incompletely understood. Here, we analyzed this interdependence by taking advantage of highly dynamic changes in expression of microRNA 181 family members miR-181a-1 and miR-181b-1 (miR-181a/b-1) during late T-cell development with very high levels of expression during thymocyte selection, followed by massive down-regulation in the periphery. Loss of miR-181a/b-1 resulted in inefficient de novo generation of Treg cells in the thymus but simultaneously permitted homeostatic expansion in the periphery in the absence of competition. Modulation of T-cell receptor (TCR) signal strength in vivo indicated that miR-181a/b-1 controlled Treg-cell formation via establishing adequate signaling thresholds. Unexpectedly, miR-181a/b-1–deficient Treg cells displayed elevated suppressive capacity in vivo, in line with elevated levels of cytotoxic T-lymphocyte–associated 4 (CTLA-4) protein, but not mRNA, in thymic and peripheral Treg cells. Therefore, we propose that intrathymic miR-181a/b-1 controls development of Treg cells and imposes a developmental legacy on their peripheral function.
Post-transcriptional gene regulation through microRNA (miRNA) has emerged as a major control mechanism of multiple biological processes, including development and function of T cells. T cells are vital components of the immune system, with conventional T cells playing a central role in adaptive immunity and unconventional T cells having additional functions reminiscent of both innate and adaptive immunity, such as involvement in stress responses and tissue homeostasis. Unconventional T cells encompass cells expressing semi-invariant T cell receptors (TCRs), such as invariant Natural Killer T (iNKT) and Mucosal-Associated Invariant T (MAIT) cells. Additionally, some T cells with diverse TCR repertoires, including γδT cells, intraepithelial lymphocytes (IEL) and regulatory T (Treg) cells, share some functional and/or developmental features with their semi-invariant unconventional counterparts. Unconventional T cells are particularly sensitive to disruption of miRNA function, both globally and on the individual miRNA level. Here, we review the role of miRNA in the development and function of unconventional T cells from an iNKT-centric point of view. The function of single miRNAs can provide important insights into shared and individual pathways for the formation of different unconventional T cell subsets.
B-cell development and function depend on stage-specific signaling through the B-cell antigen receptor (BCR). Signaling and intracellular trafficking of the BCR are connected, but the molecular mechanisms of this link are incompletely understood. Here, we investigated the role of the endosomal adaptor protein and member of the LAMTOR/Ragulator complex LAMTOR2 (p14) in B-cell development. Efficient conditional deletion of LAMTOR2 at the pre-B1 stage using mb1-Cre mice resulted in complete developmental arrest. Deletion of LAMTOR2 using Cd19-Cre mice permitted analysis of residual B cells at later developmental stages, revealing that LAMTOR2 was critical for the generation and activation of mature B lymphocytes. Loss of LAMTOR2 resulted in aberrant BCR signaling due to delayed receptor internalization and endosomal trafficking. In conclusion, we identify LAMTOR2 as critical regulator of BCR trafficking and signaling that is essential for early B-cell development in mice.
Of the hepatic cell lines developed for in vitro studies of hepatic functions as alternatives to primary human hepatocytes, many have lost major liver-like functions, but not HepaRG cells. The increasing use of the latter worldwide raises the need for establishing the reference functional status of early biobanked HepaRG cells. Using deep proteome and secretome analyses, the levels of master regulators of the hepatic phenotype and of the structural elements ensuring biliary polarity were found to be close to those in primary hepatocytes. HepaRG cells proved to be highly differentiated, with functional mitochondria, hepatokine secretion abilities, and an adequate response to insulin. Among differences between primary human hepatocytes and HepaRG cells, the factors that possibly support HepaRG transdifferentiation properties are discussed. The HepaRG cell system thus appears as a robust surrogate for primary hepatocytes, which is versatile enough to study not only xenobiotic detoxification, but also the control of hepatic energy metabolism, secretory function and disease-related mechanisms.
Background: Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as "Large Oncosomes" (LO), have been identified in highly migratory and invasive PCa cells. We recently developed and characterized the DU145R80 subline, selected from parental DU145 cells as resistant to inhibitors of mevalonate pathway. DU145R80 showed different proteomic profile compared to parental DU145 cells, along with altered cytoskeleton dynamics and a more aggressive phenotype.
Methods: Immunofluorescence staining and western blotting were used to identify blebbing and EVs protein cargo. EVs, purified by gradient ultra-centrifugations, were analyzed by tunable resistive pulse sensing and multi-parametric flow cytometry approach coupled with high-resolution imaging technologies. LO functional effects were tested in vitro by adhesion and invasion assays and in vivo xenograft model in nude mice. Xenograft and patient tumor tissues were analyzed by immunohistochemistry.
Results: We found spontaneous blebbing and increased shedding of LO from DU145R80 compared to DU145 cells. LO from DU145R80, compared to those from DU145, carried increased amounts of key-molecules involved in PCa progression including integrin alpha V (αV-integrin). By incubating DU145 cells with DU145R80-derived LO we demonstrated that αV-integrin on LO surface was functionally involved in the increased adhesion and invasion of recipient cells, via AKT. Indeed either the pre-incubation of LO with an αV-integrin blocking antibody, or a specific AKT inhibition in recipient cells are able to revert the LO-induced functional effects. Moreover, DU145R80-derived LO also increased DU145 tumor engraftment in a mice model. Finally, we identified αV-integrin positive LO-like structures in tumor xenografts as well as in PCa patient tissues. Increased αV-integrin tumor expression correlated with high Gleason score and lymph node status.
Conclusions: Overall, this study is the first to demonstrate the critical role of αV-integrin positive LO in PCa aggressive features, adding new insights in biological function of these large EVs and suggesting their potential use as PCa prognostic markers.
Evaluation of INSTAND e.V.’s external quality assessment for C-reactive protein and procalcitonin
(2019)
Background: The purpose of this paper was to analyze the general diagnostic strength and performance of in vitro diagnostics for C-reactive protein and procalcitonin based on the results of external quality assessment schemes (EQAs).
Methods: We analyzed qualitative and quantitative data on both markers collected by the Society for Promotion Quality Assurance in Medical Laboratories (INSTAND e.V.) from 20 EQAs. The C-reactive protein evaluation was method-specific and the procalcitonin evaluation manufacturer-specific (pseudonymized). Coefficients of variation were determined in order to evaluate interlaboratory comparability and the performance of individual laboratories during the analyzed period was examined.
Results: Overall most of our participants were able to correctly distinguish the positive from the negative samples, but we occasionally observed also false-positive results for the immunological detection of C-reactive protein. For the semi-quantitative results of C-reactive protein we observed an overall median difference below 5% except for dry chemistry methods (≤ 21%). For procalcitonin two manufacturer collectives showed a good comparability, while one manufacturer detected up to 42% higher results. The coefficients of variation are promising for both analytes even though they surpass the manufacturer’s indication for some collectives. The performance of individual laboratories during the analyzed period was more stable for C-reactive protein than for procalcitonin.
Conclusion: In-vitro diagnostic testing for C-reactive protein and procalcitonin showed promising results in our EQAs but still further improvements are needed. We recommend stepping up research on reference measurement methods for both parameters to possibly enhancing the accuracy and diagnostic strength of such assays.
To date, in-cell NMR has elucidated various aspects of protein behaviour by associating structures in physiological conditions. Meanwhile, current studies of this method mostly have deduced protein states in cells exclusively based on ‘indirect’ structural information from peak patterns and chemical shift changes but not ‘direct’ data explicitly including interatomic distances and angles. To fully understand the functions and physical properties of proteins inside cells, it is indispensable to obtain explicit structural data or determine three-dimensional (3D) structures of proteins in cells. Whilst the short lifetime of cells in a sample tube, low sample concentrations, and massive background signals make it difficult to observe NMR signals from proteins inside cells, several methodological advances help to overcome the problems. Paramagnetic effects have an outstanding potential for in-cell structural analysis. The combination of a limited amount of experimental in-cell data with software for ab initio protein structure prediction opens an avenue to visualise 3D protein structures inside cells. Conventional nuclear Overhauser effect spectroscopy (NOESY)-based structure determination is advantageous to elucidate the conformations of side-chain atoms of proteins as well as global structures. In this article, we review current progress for the structure analysis of proteins in living systems and discuss the feasibility of its future works.
1H-detected solid-state NMR experiments feasible at fast magic-angle spinning (MAS) frequencies allow accessing 1H chemical shifts of proteins in solids, which enables their interpretation in terms of secondary structure. Here we present 1H and 13C-detected NMR spectra of the RNA polymerase subunit Rpo7 in complex with unlabeled Rpo4 and use the 13C, 15N, and 1H chemical-shift values deduced from them to study the secondary structure of the protein in comparison to a known crystal structure. We applied the automated resonance assignment approach FLYA including 1H-detected solid-state NMR spectra and show its success in comparison to manual spectral assignment. Our results show that reasonably reliable secondary-structure information can be obtained from 1H secondary chemical shifts (SCS) alone by using the sum of 1Hα and 1HN SCS rather than by TALOS. The confidence, especially at the boundaries of the observed secondary structure elements, is found to increase when evaluating 13C chemical shifts, here either by using TALOS or in terms of 13C SCS.
In this work we provided additional insights into our understanding of bulk QCD matter through the study of the transport coeffcients which govern the non-equilibrium microscopical processes of statistical ensembles. Specically, we focused on the low energy regime corresponding to the hadron gas, as the properties of this region of the phase diagram are still relatively unknown, and existing calculations for the transport coeffcients are either scarce, contradictory, or somewhat limited in scope; this thesis' main goal was thus to shed some light on this by providing new independent calculations of these quantities.
We subsequently presented two formalisms which can be used to calculate transport coeffcients. The first one (which also was the main tool we used in the following chapters to produce our results) relies on the development of so-called Green-Kubo formulas, which relate non-equilibrium dissipative fluctuations with transport coeffcients; notably, the off-diagonal components of the energy-momentum tensor are shown to be related to the shear viscosity, its diagonal components to the bulk viscosity and fluctuations in the electric current can be related to the electric conductivity. We additionally introduced two new conductivities, namely the baryon-electric and strange electric conductivities, which we dubbed, together with the already known electric one, the "cross-conductivity", which encodes information about how electric fluctuations are correlated to changes in electric, baryonic or strange currents, or vice-versa. The second way of calculating transport coeffcient which we discussed consists in linearizing the collision term of the Boltzmann equation through the Chapman-Enskog formalism. While in principle providing direct semi-analytical results for the transport coeffcients, this approach is complicated to implement when more than a few species are considered, and as such was then mostly used as a tool to calibrate our Green-Kubo calculations.
The hadron gas model that we used for all calculations, namely the transport approach SMASH, was then presented. The main features of the model were explained, such as the collision criterion, the considered degrees of freedom and the specific way in which they microscopically interact with each other. It was verified that SMASH does reproduce analytical results of the Boltzmann equation in an expanding universe scenario, thus showing the equivalence of this transport approach and the associated kinetic theory results. A special care was taken to detail the ways in which a state of thermal and chemical equilibrium (which is necessary for Green-Kubo relations to be valid) can be reached and described using SMASH.
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Lattice QCD with heavy quarks reduces to a three-dimensional effective theory of Polyakov loops, which is amenable to series expansion methods. We analyse the effective theory in the cold and dense regime for a general number of colours, Nc. In particular, we investigate the transition from a hadron gas to baryon condensation. For any finite lattice spacing, we find the transition to become stronger, i.e. ultimately first-order, as Nc is made large. Moreover, in the baryon condensed regime, we find the pressure to scale as p ∼ Nc through three orders in the hopping expansion. Such a phase differs from a hadron gas with p ∼ N0c, or a quark gluon plasma, p ∼ N2c, and was termed quarkyonic in the literature, since it shows both baryon-like and quark-like aspects. A lattice filling with baryon number shows a rapid and smooth transition from condensing baryons to a crystal of saturated quark matter, due to the Pauli principle, and is consistent with this picture. For continuum physics, the continuum limit needs to be taken before the large Nc limit, which is not yet possible in practice. However, in the controlled range of lattice spacings and Nc-values, our results are stable when the limits are approached in this order. We discuss possible implications for physical QCD.
Alexander Ludwig: The discussion about lower delayed retirement credits in the German public pension system misses the point. Instead, it would be more important to increase both, delayed retirement credits and early retirement penalties, and to link them to the longer life expectancy of the working population.
Genetic code expansion facilitates position‐selective labeling of rna for biophysical studies
(2019)
Nature relies on reading and synthesizing the genetic code with high fidelity. Nucleic acid building blocks that are orthogonal to the canonical A‐T and G‐C base‐pairs are therefore uniquely suitable to facilitate position‐specific labeling of nucleic acids. Here, we employ the orthogonal kappa‐xanthosine‐base‐pair for in vitro transcription of labeled RNA. We devised an improved synthetic route to obtain the phosphoramidite of the deoxy‐version of the kappa nucleoside in solid phase synthesis. From this DNA template, we demonstrate the reliable incorporation of xanthosine during in vitro transcription. Using NMR spectroscopy, we show that xanthosine introduces only minor structural changes in an RNA helix. We furthermore synthesized a clickable 7‐deaza‐xanthosine, which allows to site‐specifically modify transcribed RNA molecules with fluorophores or other labels.
Background: Glial cells in the central nervous system play a key role in neuroinflammation and subsequent central sensitization to pain. They are therefore involved in the development of persistent pain. One of the main sites of interaction of the immune system with persistent pain has been identified as neuro-immune crosstalk at the glialopioid interface. The present study examined a potential association between the DNA methylation of two key players of glial/opioid intersection and persistent postoperative pain. Methods: In a cohort of 140 women who had undergone breast cancer surgery, and were assigned based on a 3year follow-up to either a persistent or non-persistent pain phenotype, the role of epigenetic regulation of key players in the glial-opioid interface was assessed. The methylation of genes coding for the Toll-like receptor 4 (TLR4) as a major mediator of glial contributions to persistent pain or for the μ-opioid receptor (OPRM1) was analyzed and its association with the pain phenotype was compared with that conferred by global genome-wide DNA methylation assessed via quantification of the methylation in the retrotransposon LINE1. Results: Training of machine learning algorithms indicated that the global DNA methylation provided a similar diagnostic accuracy for persistent pain as previously established non-genetic predictors. However, the diagnosis can be based on a single DNA based marker. By contrast, the methylation of TLR4 or OPRM1 genes could not contribute further to the allocation of the patients to the pain-related phenotype groups. Conclusions: While clearly supporting a predictive utility of epigenetic testing, the present analysis cannot provide support for specific epigenetic modulation of persistent postoperative pain via methylation of two key genes of the glial-opioid interface.
We report on the observation of coherent terahertz (THz) emission from the quasi-one-dimensional charge-density wave (CDW) system, blue bronze (K0.3MoO3), upon photo-excitation with ultrashort near-infrared optical pulses. The emission contains a broadband, low-frequency component due to the photo-Dember effect, which is present over the whole temperature range studied (30–300 K), as well as a narrow-band doublet centered at 1.5 THz, which is only observed in the CDW state and results from the generation of coherent transverse-optical phonons polarized perpendicular to the incommensurate CDW b-axis. As K0.3MoO3 is centrosymmetric, the lowest-order generation mechanism which can account for the polarization dependence of the phonon emission involves either a static surface field or quadrupolar terms due to the optical field gradients at the surface. This phonon signature is also present in the ground-state conductivity, and decays in strength with increasing temperature to vanish above $T\sim 100\,{\rm{K}}$, i.e. significantly below the CDW transition temperature. The temporal behavior of the phonon emission can be well described by a simple model with two coupled modes, which initially oscillate with opposite polarity.
Background: Hepatitis B (HepB) is a major public health concern in Malaysia yet little is known about knowledge and awareness of this infection in the country. Such information is essential for designing effective intervention strategies for HepB prevention and control. The aim of this study was to characterize knowledge and awareness regarding HepB in Malaysia and to identify their associated sociodemographic determinants.
Methods: A community-based cross-sectional survey was conducted between January and May 2016 in Selangor state of Malaysia. A two-stage cluster random sampling design was used and one adult member of selected households was interviewed face-to-face. Logistic regression was used to estimate the differences in knowledge and awareness between groups.
Results: A total of 764 households completed the interviews and were included in the final analysis. Only 36.9 and 38.8% of the participants had good knowledge and awareness, respectively. The factors associated with good knowledge were being in the 35–44 year age group, Malay ethnicity, high educational attainment and high family income. Being Chinese, being older and having high educational attainment were determinants of having good awareness towards HepB. Participants who had good knowledge were 2.5 times more likely to also have good awareness (OR: 2.41, 95% CI: 1.78–3.26, p < 0.001).
Conclusions: This study reveals a low level of knowledge and awareness of HepB among households in Malaysia. This finding highlights the need to improve public knowledge and awareness through well-designed programs targeting vulnerable groups in order to reduce hepatitis B virus transmission and achieve the governmental target of eliminating viral hepatitis as a public health concern by 2030.
PfEMP1 (erythrocyte membrane protein 1) adhesins play a pivotal role in the pathophysiology of falciparum malaria, by mediating sequestration of Plasmodium falciparum-infected erythrocytes in the microvasculature. PfEMP1 variants are expressed by var genes and are presented on membrane elevations, termed knobs. However, the organization of PfEMP1 on knobs is largely unclear. Here, we use super-resolution microscopy and genetically altered parasites expressing a modified var2csa gene in which the coding sequence of the photoactivatable mEOS2 was inserted to determine the number and distribution of PfEMP1 on single knobs. The data were verified by quantitative fluorescence-activated cell sorting analysis and immuno-electron microscopy together with stereology methods. We show that knobs contain 3.3 ± 1.7 and 4.3 ± 2.5 PfEMP1 molecules, predominantly placed on the knob tip, in parasitized erythrocytes containing wild type and sickle haemoglobin, respectively. The ramifications of our findings for cytoadhesion and immune evasion are discussed.
Objective: Excessive inflammation in the central nervous system (CNS) and the periphery can result in neurodegeneration and parkinsonism. Recent evidence suggests that immune responses in Parkinson disease patients are dysregulated, leading to an increased inflammatory reaction to unspecific triggers. Although α‐synuclein pathology is the hallmark of Parkinson disease, it has not been investigated whether pathologic α‐synuclein is a specific trigger for excessive inflammatory responses in Parkinson disease.
Methods: We investigated the immune response of primary human monocytes and a microglial cell line to pathologic forms of α‐synuclein by assessing cytokine release upon exposure.
Results: We show that pathologic α‐synuclein (mutations, aggregation) results in a robust inflammatory activation of human monocytes and microglial BV2 cells. The activation is conformation‐ dependent, with increasing fibrillation and early onset mutations having the strongest effect on immune activation. We also found that activation of immune cells by extracellular α‐synuclein is potentiated by extracellular vesicles, possibly by facilitating the uptake of α‐synuclein. Blood extracellular vesicles from Parkinson disease patients induce a stronger activation of monocytes than blood extracellular vesicles from healthy controls. Most importantly, monocytes from Parkinson disease patients are dysregulated and hyperactive in response to stimulation with pathologic α‐synuclein. Furthermore, we demonstrate that α‐synuclein pathology in the CNS is sufficient to induce the monocyte dysregulation in the periphery of a mouse model.
Interpretation: Taken together, our data suggest that α‐synuclein pathology and dysregulation of monocytes in Parkinson disease can act together to induce excessive inflammatory responses to α‐synuclein. ANN NEUROL 2019;86:593–606
Background: The aim of this study was to assess the knowledge and attitudes towards pregnancy-related issues of Zika virus (ZIKV) infection among general practitioners (GPs), a frontline healthcare worker group, in Indonesia.
Methods: A cross-sectional, online survey assessing knowledge and attitudes towards ZIKV infection on multiple-item scales was sent to GPs in the Sumatra and Java islands of Indonesia. The associations between independent factors and either knowledge or attitude were assessed with logistic regressions. The correlation and association between knowledge and attitude were estimated.
Results: We included 457 (53.7%) out of 850 responses in the analysis. Among these, 304 (66.5%) and 111 (24.2%) respondents had a good knowledge and attitude, respectively. No demographic, workplace, professional development, or experiential characteristics related to ZIKV infection were associated with knowledge. In the multivariate analysis, only contact experience was associated with attitude. There was a significant, positive correlation between knowledge and attitude scores.
Conclusions: Although knowledge of pregnancy-related complications of ZIKV infection is relatively high among GPs in Indonesia, more than 75% of them had a poor attitude towards pregnancy-related issues of Zika. Strategies for enhancing the capacity of GPs to develop positive attitudes and respond to ZIKV infection are needed.
Light-matter interaction in the strong coupling regime is of profound interest for fundamental quantum optics, information processing and the realization of ultrahigh-resolution sensors. Here, we report a new way to realize strong light-matter interaction, by coupling metamaterial plasmonic "quasi-particles" with photons in a photonic cavity, in the terahertz frequency range. The resultant cavity polaritons exhibit a splitting which can reach the ultra-strong coupling regime, even with the comparatively low density of quasi-particles, and inherit the high Q-factor of the cavity despite the relatively broad resonances of the Swiss-cross and split-ring-resonator metamaterials used. We also demonstrate nonlocal collective interaction of spatially separated metamaterial layers mediated by the cavity photons. By applying the quantum electrodynamic formalism to the density dependence of the polariton splitting, we can deduce the intrinsic transition dipole moment for single-quantum excitation of the metamaterial quasi-particles, which is orders of magnitude larger than those of natural atoms. These findings are of interest for the investigation of fundamental strong-coupling phenomena, but also for applications such as ultra-low-threshold terahertz polariton lasing, voltage-controlled modulators and frequency filters, and ultra-sensitive chemical and biological sensing.
Surface plasmon polaritons on (silver) nanowires are promising components for future photonic technologies. Here, we study near-field patterns on silver nanowires with a scattering-type scanning near-field optical microscope that enables the direct mapping of surface waves. We analyze the spatial pattern of the plasmon signatures for different excitation geometries and polarization and observe a plasmon wave pattern that is canted relative to the nanowire axis, which we show is due to a superposition of two different plasmon modes, as supported by electromagnetic simulations including the influence of the substrate. These findings yield new insights into the excitation and propagation of plasmon polaritons for applications in nanoplasmonic devices.
Ampharete oculicirrata sp. nov. (Annelida: Ampharetidae) is described from samples collected by the Joint Nature Conservation Committee and Marine Scotland Science, in the West Shetland Shelf NCMPA in the NE Atlantic. This species is characterised by a very small body size, thin and slender paleae, twelve thoracic and eleven abdominal uncinigers, presence of eyes both in the prostomium and the pygidium, the latter provided with a pair of long lateral cirri. The external micro-morphology of the new taxon was studied using scanning electron microscopy and compared with species described or reported from the North Atlantic. Two complementary keys to all species of Ampharete in the area are also provided.
Rhabdomyosarcoma (RMS) is the most frequent pediatric soft-tissue sarcoma comprising two major subtypes – the alveolar and the embryonal rhabdomyosarcoma. The current therapeutic regime is multimodal including surgery, radiation and chemotherapy with cytostatic drugs. Although the prognosis for RMS patients has steadily improved to a 5-year overall survival rate of 70% for ERMS and 50% for ARMS, prognosis for subgroups with primary metastases or relapsed patients is still less than 25%, highlighting the need for development of new therapies for these subgroups. Since cancer cells are addicted to their cancer promoting transcriptional program, remodeling transcription by targeting bromodomain and extraterminal (BET) proteins has emerged as compelling anticancer strategy. However, in many cancer types BET inhibition was proved cytostatic but not cytotoxic emphasizing the need for combination protocols.
In this study we identify a novel synergistic interaction of the BET inhibitor JQ1 with p110α-isoform-specific Phosphoinositid-3-Kinase (PI3K) inhibitor BYL719 (Alpelisib) to induce mitochondrial apoptosis and global reallocation of BRD4 to chromatin. At first, we showed that JQ1 single treatment had cytostatic effects at nanomolar concentrations and inhibited MYC and Hedgehog (Hh) signaling in RMS known to promote proliferation of RMS. However, JQ1 single treatment barely induced cell death in RMS cells even at concentrations of up to 20 µM (< 20% cell death). Thus, we next tested combination approaches to elicit cell death. Since we previously identified synergistic cell death induction of Hh inhibition and PI3K inhibition in RMS cells we tested JQ1 in combination with the pan-PI3K/mTOR inhibitor PI-103 and the p110α-isoform-specific PI3K inhibitor BYL719. In addition, we tested JQ1 in combination with distinct HDAC inhibitors namely JNJ-26481585, SAHA (Vorinostat), MS-275 (Entinostat) and LBH-589 (Panobinostat) since the synergistic interaction of BET and HDAC inhibition has previously been described for other tumor entities.
Interestingly the synergism of cell death induction of JQ1/BYL719 co-treatment is superior to the synergism of JQ1 with pan-PI3K/mTOR inhibitor PI-103 or the tested HDAC inhibitors as confirmed by calculation of combination index. To investigate the molecular mechanisms underlying the synergy of JQ1/BYL719 co-treatment, we performed RNA-Seq and BRD4 ChIP-Seq experiments. RNA-Seq exhibited, that JQ1/BYL719 co-treatment shifted the overall balance of BCL-2 family gene expression towards apoptosis and increased gene expression of proapoptotic BMF, BCL2L11 (BIM) and PMAIP1 (NOXA) while decreasing gene expression of antiapoptotic BCL2L1 (BCL xL). These changes were verified by qRT-PCR and Western blot. Notably, BRD4 is phosphorylated upon JQ1/BYL719 co-treatment and globally reallocates BRD4 to chromatin. This BRD4 reallocation includes enrichment of BRD4 at the super-enhancer site of BMF, at the super-enhancer, typical enhancer and promoter regions of BCL2L11 (BIM) and at the PMAIP1 (NOXA) promoter, while JQ1 alone, as expected, reduces global chromatin binding of BRD4. Integration of RNA-Seq and BRD4 ChIP-Seq data underlines the transcriptional relevance of reallocated BRD4 upon JQ1/BYL719 co-treatment. Immunopreciptation studies showed, that RMS cells are initially primed to undergo mitochondrial apoptosis since BIM is constitutively bound to antiapoptotic BCL-2, BCL xL and MCL-1. JQ1/BYL719 co-treatment increased BIM expression and its neutralization of antiapoptotic BCL-2, BCL-xL and MCL-1 thereby rebalancing the ratio of pro- and antiapoptotic BCL-2 proteins in favor of apoptosis. This promotes activation of BAK and BAX resulting in caspase-dependent apoptosis. The functional relevance of proapoptotic re-balancing for the execution of JQ1/BYL719-mediated apoptosis was confirmed by individual silencing of BMF, BIM, NOXA or overexpression of BCL-2 or MCL-1, which all significantly rescued JQ1/BYL719-induced cell death. Execution of cell death by mitochondrial caspase-dependent apoptosis was veryfied by individual knockdown of BAK and BAX or caspase inhibitor N-Benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethylketone (zVAD.fmk), which all significantly rescued JQ1/BYL719-induced cell death.
In summary, combined BET and PI3Kα inhibition cooperatively induces mitochondrial apoptosis by proapoptotic re-balancing of BCL-2 family proteins accompanied by reallocation of BRD4 to transcriptional regulatory elements of BH3-only proteins.
Demographic change is supposed to be the most important indirect driver for changing biodiversity. In this article, a systematic review of 148 studies was conducted to examine the scientific evidence for this relationship and to identify potential gaps in research. We explored the spatial distribution of studies, the categories addressed with respect to biodiversity and demographic change, and the ways in which their relationships were conceptualised (spatially and temporally) and valued. The majority of studies were carried out in Africa, Europe and North America. Our analysis confirms the trend that demographic phenomena were mostly found to negatively influence biodiversity. However, a considerable number of studies also point towards impacts that were context dependent, either positive or negative under certain circumstances. In addition to that we identified significant gaps in research. In particular, there is a lack of addressing (1) other demographic aspects such as population decline, age structure or gender differences, (2) spatial variability of, e.g. human population growth, (3) long-term effects of demographic processes, and (4) the context dependency (e.g. regulations/law enforcement, type of human activities, and choice of scale or proxy). We conclude there is evidence that the relationship between biodiversity and demographic change is much more complex than expected and so far represented in research. Thus, we call for a social–ecological biodiversity research that particularly focusses on the functional relation between biodiversity and human activities, namely the different types, context, and interdependent dynamics (spatial and temporal) of this complex relation.
A machine-learned analysis suggests non-redundant diagnostic information in olfactory subtests
(2019)
Background: The functional performance of the human sense of smell can be approached via assessment of the olfactory threshold, the ability to discriminate odors or the ability to identify odors. Contemporary clinical test batteries include all or a selection of these components, with some dissent about the required number and choice.
Methods: Olfactory thresholds, odor discrimination and odor identification scores were available from 10,714 subjects (3662 with anomia, 4299 with hyposmia, and 2752 with normal olfactory function). To assess, whether the olfactory subtests confer the same information or each subtest confers at least partly non-redundant information relevant to the olfactory diagnosis, we compared the diagnostic accuracy of supervised machine learning algorithms trained with the complete information from all three subtests with that obtained when performing the training with the information of only two or one subtests.
Results: The training of machine-learned algorithms with the full information about olfactory thresholds, odor discrimination and odor identification from 2/3 of the cases, resulted in a balanced olfactory diagnostic accuracy of 98% or better in the 1/3 remaining cases. The most pronounced decrease in the balanced accuracy, to approximately 85%, was observed when omitting olfactory thresholds from the training, whereas omitting odor discrimination or identification was associated with smaller decreases (balanced accuracies approximately 90%).
Conclusions: Results support partly non-redundant contributions of each olfactory subtest to the clinical olfactory diagnosis. Olfactory thresholds provided the largest amount of non-redundant information to the olfactory diagnosis.
The ubiquitin-related SUMO system represents a versatile post-translational modification pathway controlling a variety of cellular signalling networks. In mammalian cells, lysine residues of target proteins can be covalently modified with three SUMO isoforms (SUMO1, SUMO2 and SUMO3) resulting in conjugation of either single SUMO moieties or formation of poly-SUMO chains. Importantly, SUMO modification is a reversible process, where the deconjugation of SUMO from its substrates is mediated by SUMO proteases. In humans, the best-characterized subfamily is the SENP family of SUMO-specific isopeptidases comprised of SENP1-3 and SENP5-7. For undisturbed cellular signalling events, a proper balance of SUMO conjugation and deconjugation is crucial. SENPs fulfil the important function of counteracting SUMOylation. A key question is how the relatively low number of SENPs specifically controls the SUMOylation status of hundreds of cellular proteins.
The aim of this thesis was to uncover the regulation and substrate specificity of distinct SUMO isopeptidases in order to better understand their role in cellular signalling pathways.
In the first part of this work, we investigated the influence of hypoxia on SUMO signalling, in particular on the activity of SENPs. Importantly, we found that the catalytic activity of distinct SENPs (especially SENP1 and SENP3) is strongly but reversibly diminished under low oxygen. As a consequence, the SUMO modification of a specific subset of proteins is changed under hypoxia. We specifically identified proteins being hyperSUMOylated after 24 hours of hypoxia by SUMO1 immunoprecipitation followed by mass spectrometry. We further validated the transcriptional co-repressor BHLHE40 as hypoxic SUMO target and confirmed SENP1 as responsible isopeptidase for deconjugation of SUMOylated BHLHE40. We provide evidence that SUMO conjugation to BHLHE40 enhances its repressive functions on the expression of the metabolic master regulator PGC-1α. Therefore we propose a model where inactivation of SENP1 under hypoxia results in SUMOylated BHLHE40, possibly contributing to metabolic reprogramming under hypoxia.
To get insight into substrate selectivity of SENP family members, in particular SENP3 and SENP6, we choose a proteomic profiling strategy. For the identification of specific SUMO substrates controlled by SENP3, we applied a large-scale IP-MS approach in SENP3 KO and WT cells. The most strongly induced SUMO targets in the absence of SENP3 were key regulators of ribosome maturation. We identified factors involved in the remodelling of both 90S and 60S pre-ribosomes. SENP3 has already been described as being critically involved in maturation of the pre-60S subunit and 28S rRNA processing. Previously described SENP3-regulated master targets in this process are the ribosome maturation factors PELP1 and Las1L. Importantly, both were also identified as the most significantly regulated SENP3 targets in our unbiased proteomic approach. Importantly, however, enhanced SUMOylation was also detected on 90S-associated regulators, such as BMS1. Altogether, these data strengthen the functional link between SENP3 and ribosome biogenesis and point to a role of SENP3 beyond 60S maturation.
In addition to SENP3, we explored the substrate specificity of SENP6, which mainly acts on polymeric SUMO2/3 chains. Applying a proteomic profiling strategy, we were able to identify SENP6-controlled SUMO networks functioning in DNA damage response as well as chromatin organization. We demonstrated that SENP6 reverses polySUMOylation of several subunits of the cohesin complex, thereby regulating the SUMOylation status and chromatin association of this complex. Furthermore, we found a tight interaction of SENP6 with the hPSO4/PRP19 complex, involved in DNA damage response by activation of the ATR-CHK1 signalling cascade. In cells depleted of SENP6, we observe deficient recruitment of the co-activator ATRIP to chromatin which results in diminished CHK1 activation. We therefore illustrate a general role of SENP6 in the control of chromatin-associated protein networks involved in genome integrity and chromatin organization.
Enolase is a glycolytic enzyme, which catalyzes the inter-conversion of 2-phosphoglycerate to phosphoenolpyruvate. Altered expression of this enzyme is frequently observed in cancer and accounts for the Warburg effect, an adaptive response of tumor cells to hypoxia. In addition to its catalytic function, ENO-1 exhibits other activities, which strongly depend on its cellular and extracellular localization. For example, the association of ENO-1 with mitochondria membrane was found to be important for the stability of the mitochondrial membrane, and ENO-1 sequestration on the cell surface was crucial for plasmin-mediated pericellular proteolysis. The latter activity of ENO-1 enables many pathogens but also immune and cancer cells to invade the tissue, leading further to infection, inflammation or metastasis formation. The ability of ENO-1 to conduct so many diverse processes is reflected by its contribution to a high number of pathologies, including type 2 diabetes, cardiovascular hypertrophy, fungal and bacterial infections, cancer, systemic lupus erythematosus, hepatic fibrosis, Alzheimer’s disease, rheumatoid arthritis, and systemic sclerosis. These unexpected non-catalytic functions of ENO-1 and their contributions to diseases are the subjects of this review.
Background: Pediatric community acquired pneumonia (pedCAP) is one of the leading causes for childhood morbidity accounting for up to 20% of pediatric hospital admissions in high income countries. In spite of its high morbidity, updated epidemiological and pathogen data after introduction of preventive vaccination and novel pathogen screening strategies are limited. Moreover, there is a need for validated recommendations on diagnostic and treatment regimens in pedCAP. Through collection of patient data and analysis of pathogen and host factors in a large sample of unselected pedCAP patients in Germany, we aim to address and substantially improve this situation.
Methods: pedCAPNETZ is an observational, multi-center study on pedCAP. Thus far, nine study centers in hospitals, outpatient clinics and practices have been initiated and more than 400 patients with radiologically confirmed pneumonia have been enrolled, aiming at a total of 1000 study participants. Employing an online data base, information on disease course, treatment as well as demographical and socioeconomical data is recorded. Patients are followed up until day 90 after enrollment; Comprehensive biosample collection and a central pedCAPNETZ biobank allow for in-depth analyses of pathogen and host factors. Standardized workflows to assure sample logistics and data management in more than fifteen future study centers have been established.
Discussion: Through comprehensive epidemiological, clinical and biological analyses, pedCAPNETZ fills an important gap in pediatric and infection research. To secure dissemination of the registry, we will raise clinical and scientific awareness at all levels. We aim at participating in decision making processes for guidelines and prevention strategies. Ultimately, we hope the results of the pedCAPNETZ registry will help to improve care and quality of life in pedCAP patients in the future.
By running a temperature series of molecular dynamics (MD) simulations starting from the known low-temperature phase, the experimentally observed phase transition in a `jumping crystal' was captured, thereby providing a prediction of the unknown crystal structure of the high-temperature phase and clarifying the phase-transition mechanism. The phase transition is accompanied by a discontinuity in two of the unit-cell parameters. The structure of the high-temperature phase is very similar to that of the low-temperature phase. The anisotropic displacement parameters calculated from the MD simulations readily identified libration as the driving force behind the phase transition. Both the predicted crystal structure and the phase-transition mechanism were verified experimentally using TLS (translation, libration, screw) refinement against X-ray powder diffraction data.
We have encountered two polymorphs of the title compound, C24H16B2OS2, both of which display almost the same unit-cell parameters. Compound (I) crystallizes in the non-centrosymmetric space group P21 with four molecules in the asymmetric unit. These molecules are related by pseudosymmetry. As a result, the space group looks like P21/c, but the structure cannot be refined successfully in that space group. Compound (II) on the other hand crystallizes in the centrosymmetric space group P21/c with only two molecules in the asymmetric unit. The crystals studied for (I) and (II) were both non-merohedral twins.
Iodo(triphenyl)silane
(2019)
The molecular structure of the title compound, C18H15ISi, which crystallizes in the space group C2/c, does not exhibit any unusual features. Two weak C—H⋯π interactions may help to consolidate the packing. The present structure is not isostructural with the known Ph3SiX (X = F, Cl or Br) compounds.
Men and women differ in their vulnerability to a variety of stress-related illnesses, but the underlying neurobiological mechanisms are not well understood. This is likely due to a comparative dearth of neurobiological studies that assess male and female rodents at the same time, while human neuroimaging studies often don’t model sex as a variable of interest. These sex differences are often attributed to the actions of sex hormones, i.e. estrogens, progestogens and androgens. In this review, we summarize the results on sex hormone actions in the hippocampus and seek to bridge the gap between animal models and findings in humans. However, while effects of sex hormones on the hippocampus are largely consistent in animals and humans, methodological differences challenge the comparability of animal and human studies on stress effects. We summarise our current understanding of the neurobiological mechanisms that underlie sex-related differences in behavior and discuss implications for stress-related illnesses.
Background: The effects of blood flow restriction (training) may serve as a model of peripheral artery disease. In both conditions, circulating micro RNAs (miRNAs) are suggested to play a crucial role during exercise-induced arteriogenesis. We aimed to determine whether the profile of circulating miRNAs is altered after acute resistance training during blood flow restriction (BFR) as compared with unrestricted low- and high-volume training, and we hypothesized that miRNA that are relevant for arteriogenesis are affected after resistance training.
Methods: Eighteen healthy volunteers (aged 25 ± 2 years) were enrolled in this three-arm, randomized-balanced crossover study. The arms were single bouts of leg flexion/extension resistance training at (1) 70% of the individual single-repetition maximum (1RM), (2) at 30% of the 1RM, and (3) at 30% of the 1RM with BFR (artificially applied by a cuff at 300 mm Hg). Before the first exercise intervention, the individual 1RM (N) and the blood flow velocity (m/s) used to validate the BFR application were determined. During each training intervention, load-associated outcomes (fatigue, heart rate, and exhaustion) were monitored. Acute effects (circulating miRNAs, lactate) were determined using pre-and post-intervention measurements.
Results: All training interventions increased lactate concentration and heart rate (p < 0.001). The high-intensity intervention (HI) resulted in a higher lactate concentration than both lower-intensity training protocols with BFR (LI-BFR) and without (LI) (LI, p = 0.003; 30% LI-BFR, p = 0.008). The level of miR-143-3p was down-regulated by LI-BFR, and miR-139-5p, miR-143-3p, miR-195-5p, miR-197-3p, miR-30a-5p, and miR-10b-5p were up-regulated after HI. The lactate concentration and miR-143-3p expression showed a significant positive linear correlation (p = 0.009, r = 0.52). A partial correlation (intervention partialized) showed a systematic impact of the type of training (LI-BFR vs. HI) on the association (r = 0.35 remaining after partialization of training type).
Conclusions: The strong effects of LI-BFR and HI on lactate- and arteriogenesis-associated miRNA-143-3p in young and healthy athletes are consistent with an important role of this particular miRNA in metabolic processes during (here) artificial blood flow restriction. BFR may be able to mimic the occlusion of a larger artery which leads to increased collateral flow, and it may therefore serve as an external stimulus of arteriogenesis.
Members of conflicting groups experience threats to different identity dimensions, resulting in the need to restore the aspect of identity that was threatened. Do these needs translate into specific goals in social interactions? In the present research, we examined the hypotheses that (1) experiencing one’s ingroup as illegitimately disadvantaged or victimized arouses agentic goals (to act and appear assertive and confident) when interacting with the advantaged or victimizing group, while (2) experiencing one’s ingroup as illegitimately advantaged or perpetrating transgressions arouses communal goals (to act and appear warm and trustworthy) when interacting with the disadvantaged or victimized group. Study 1 (N = 391) generally supported both hypotheses across diverse intergroup contexts involving gender, national/ethnic, and consumer identities. Study 2 (N = 122) replicated this pattern in a context of occupational identities. Study 2 further showed that the effect of ingroup role on agentic and communal intergroup goals was not moderated by participants’ general dispositional preferences for agentic and communal goals in interpersonal interactions, thus demonstrating how ingroup role exerts a distinct and robust influence on goals for interactions with other groups. Theoretical and practical implications are discussed.
Among the causality assessment methods used for the diagnosis of drug-induced liver injury (DILI), Roussel Uclaf Causality Assessment Method (RUCAM) remains the most widely used not only for individual cases but also for prospective and retrospective studies worldwide. This first place is justified by the characteristics of the method such as precise definition and classification of the liver injury, which determines the right scale in the scoring system, precise definition of the seven criteria, and the validation approach based on cases with positive rechallenge. RUCAM is used not only for any types of drugs but also for herbal medicines causing herb-induced liver injury, (HILI) and dietary supplements. In 2016, the updated RUCAM provided further specifications of criteria and instructions to improve interobserver variability. Although this method was criticized for criteria such as the age and alcohol consumption, recent consensus meeting of experts has recognized their value and recommended their incorporation into any method. While early studies searching for DILI in large databases especially in electronic medical records were based on codes of diseases or natural language without causality assessment, the recommendation is now to include RUCAM in the search for DILI/HILI. There are still studies on DILI detection or the identification of biomarkers that take into consideration the cases assessed as “possible,” although it is well known that these cases reduce the strength of the association between the cases and the offending compound or the new biomarker to be validated. Attempts to build electronic RUCAM or automatized application of this method were successful despite some weaknesses to be corrected. In the future, more reflections are needed on an expert system to standardize the exclusion of alternative causes according to the clinical context. Education and training on RUCAM should be encouraged to improve the results of the studies and the day-to-day work in pharmacovigilance departments in companies or in regulatory agencies. It is also expected to improve RUCAM with biomarkers or other criteria provided that the validation process replaces expert opinion by robust standards such as those used for the original method.
Influence of structured reporting of tooth-colored indirect restorations on clinical decision-making
(2019)
The aim of the present study was to discover what influence structured reporting (study group = A) of toothcoloured lab-fabricated restorations has on clinical decision-making following international guidelines. By way of comparison, the conventional approach in the form of short reporting with 5 items (control group = B) was used as gold standard. The study was carried out in the first clinical semester of dentistry (n = 68) at the Goethe University in Frankfurt am Main. In the study group, indirect ceramic restorations were assessed on a scale of 1 (very good) to 5 (insufficient) using structured reporting (7 items, each with 5 subgroups) in accordance to World Dental Federation (FDI) - standards. Following this, the clinical decision on the insertion of the restoration was made. To evaluate the quality of the structured reporting, sensitivity, specificity, confidence intervals (Cl) and the respective predictive values (positive = PPV, negative= NPV) were determined. Based on FDI reporting, a ceramic inlay is also favored with a great degree of certainty in clinical decisions: this was the true in 34 procedures out of a total of 38 clinically incorporated ceramic inlays [sensitivity 67% (95% CI: 46%83%); specificity 89% (95% Cl: 75%-97%); PPV 82%, NPV 79%]. In the control group, sensitivity was 56% (95% CI: 35%-75%); specificity 92% (95% CI: 79%-98%); PPV 83%, NPV 74%. No significant differences could be determined between A and B (p = 0.813).
Due to the higher sensitivity and efficiency given comparable specificity, structured reporting of tooth-coloured lab-fabricated restorations based on FDI criteria, appears more recommendable than short reporting. It is also suitable for promoting decision-making in quality assessment, thus improving the durability of dental restorations.
Strong dose response after immunotherapy with PQ grass using conjunctival provocation testing
(2019)
Background: Pollinex Quattro Grass (PQ Grass) is an effective, well-tolerated, short pre-seasonal subcutaneous immunotherapy to treat seasonal allergic rhinoconjunctivitis (SAR) due to grass pollen. In this Phase II study, 4 cumulative doses of PQ Grass and placebo were evaluated to determine its optimal cumulative dose.
Methods: Patients with grass pollen-induced SAR were randomised to either a cumulative dose of PQ Grass (5100, 14400, 27600 and 35600 SU) or placebo, administered as 6 weekly subcutaneous injections over 31–41 days (EudraCT number 2017-000333-31). Standardized conjunctival provocation tests (CPT) using grass pollen allergen extract were performed at screening, baseline and post-treatment to determine the total symptom score (TSS) assessed approximately 4 weeks after dosing. Three models were pre-defined (Emax, logistic, and linear in log-dose model) to evaluate a dose response relationship.
Results: In total, 95.5% of the 447 randomized patients received all 6 injections. A highly statistically significant (p < 0.0001), monotonic dose response was observed for all three pre-specified models. All treatment groups showed a statistically significant decrease from baseline in TSS compared to placebo, with the largest decrease observed after 27600 SU (p < 0.0001). The full course of 6 injections was completed by 95.5% of patients. Treatment-emergent adverse events were similar across PQ Grass groups, and mostly mild and transient in nature.
Conclusions: PQ Grass demonstrated a strong curvilinear dose response in TSS following CPT without compromising its safety profile.
Background: Chronic hepatitis C is a major public health burden. With new interferon-free direct-acting agents (showing sustained viral response rates of more than 98%), elimination of HCV seems feasible for the first time. However, as HCV infection often remains undiagnosed, screening is crucial for improving health outcomes of HCV-patients. Our aim was to assess the long-term cost-effectiveness of a nationwide screening strategy in Germany.
Methods: We used a Markov cohort model to simulate disease progression and examine long-term population outcomes, HCV associated costs and cost-effectiveness of HCV screening. The model divides the total population into three subpopulations: general population (GEP), people who inject drugs (PWID) and HIV-infected men who have sex with men (MSM), with total infection numbers being highest in GEP, but new infections occurring only in PWIDs and MSM. The model compares four alternative screening strategies (no/basic/advanced/total screening) differing in participation and treatment rates.
Results: Total number of HCV-infected patients declined from 275,000 in 2015 to between 125,000 (no screening) and 14,000 (total screening) in 2040. Similarly, lost quality adjusted life years (QALYs) were 320,000 QALYs lower, while costs were 2.4 billion EUR higher in total screening compared to no screening. While incremental cost-effectiveness ratio (ICER) increased sharply in GEP and MSM with more comprehensive strategies (30,000 EUR per QALY for total vs. advanced screening), ICER decreased in PWIDs (30 EUR per QALY for total vs. advanced screening).
Conclusions: Screening is key to have an efficient decline of the HCV-infected population in Germany. Recommendation for an overall population screening is to screen the total PWID subpopulation, and to apply less comprehensive advanced screening for MSM and GEP.
The paper outlines a method for investigating the speed effect due to a time limit in testing. It is assumed that the time limit enables latent processing speed to influence responses by causing omissions in the case of insufficient speed. Because of processing speed as additional latent source, the customary confirmatory factor model is enlarged by a second latent variable representing latent processing speed. For distinguishing this effect from other method effects, the factor loadings are fixed according to the cumulative normal distribution. With the second latent variable added, confirmatory factor analysis of reasoning data (N=518) including omissions because of a time limit yielded good model fit and discriminated the speed effect from other possible effects due to the item difficulty, the homogeneity of an item subset and the item positions. Because of the crucial role of the cumulative normal distribution for fixing the factor loadings a check of the normality assumption is also reported.
Post‐traumatic stress disorder (PTSD) is associated with a hypersensitivity to potential threat. This hypersensitivity manifests through differential patterns of emotional information processing and has been demonstrated in behavioral and neurophysiological experimental paradigms. However, the majority of research has been focused on adult patients with PTSD. To examine possible differences in underlying neurophysiological patterns for adolescent patients with PTSD after childhood sexual and/or physical abuse (CSA/CPA), ERP correlates of emotional word processing in 38 healthy participants and 40 adolescent participants with PTSD after experiencing CSA/CPA were studied. The experimental paradigm consisted of a passive reading task with neutral, positive (e.g., paradise), physically threatening (e.g., torment), and socially threatening (i.e., swearing, e.g., son of a bitch) words. A modulation of P3 amplitudes by emotional valence was found, with positive words inducing less elevated amplitudes over both groups. Interestingly, in later processing, the PTSD group showed augmented early late positive potential (LPP) amplitudes for socially threatening stimuli, while there were no modulations within the healthy control group. Also, region‐specific emotional modulations for anterior and posterior electrode clusters were found. For the anterior LPP, highest activations have been found for positive words, while socially and physically threatening words led to strongest modulations in the posterior LPP cluster. There were no modulations by group or emotional valence at the P1 and EPN stage. The findings suggest an enhanced conscious processing of socially threatening words in adolescent patients with PTSD after CSA/CPA, pointing to the importance of a disjoined examination of threat words in emotional processing research.
Cardiac reactions to emotional words in adolescents and young adults with PTSD after child abuse
(2019)
Post‐traumatic stress disorder (PTSD) is associated with alterations in cardiac reactivity to threat cues. Meta‐analyses have summarized that adults with PTSD have increased heart rates in response to trauma‐related stimuli. However, the opposite effect (i.e., cardiac hyporeactivity) has recently been reported in subgroups of PTSD patients. In children and adolescents with PTSD, reports of cardiac alterations are rare and ambiguous. So far, most studies in adolescents and young adults are restricted to victims of accidents, even though PTSD is highly prevalent in victims of child maltreatment. The present study aimed at investigating cardiac reactions in adolescents and young adults with PTSD after child abuse. Cardiac responses to standardized emotional words were studied in 39 adolescent and young adult PTSD patients after childhood sexual and/or physical abuse as compared to 39 healthy control subjects (age range: 15–20 years). The experimental paradigm consisted of a passive reading task with neutral, positive, physically threatening, and socially threatening (swear) words. Results showed that cardiac reactions to negative stimuli, particularly physically threatening stimuli, were less pronounced in PTSD patients than in controls. Moreover, cardiac reactions in response to socially threatening words were less variable in the PTSD group. No differences between and within groups were present in reaction to neutral or positive stimuli. Findings suggest that a physiologically blunted subtype of PTSD may already manifest during adolescence and young adulthood. Moreover, the results of the present study emphasize the relevance of individual trauma history for physiological reactions.
Background: Synovial fibroblasts (SF) play a major role in the pathogenesis of rheumatoid arthritis (RA) and develop an aggressive phenotype destroying cartilage and bone, thus termed RASF. JAK inhibitors have shown to be an efficient therapeutic option in RA treatment, but less is known about the effect of JAK inhibitors on activated RASF. The aim of the study was to examine the effects of JAK inhibitors on activated RASF.
Methods: Synovium of RA patients was obtained during knee replacement surgeries. Synoviocytes were isolated and pretreated with JAK inhibitors. Pro-inflammatory cytokines and matrix degrading proteinases were measured by ELISA in supernatant after stimulation with oncostatin M or IL-1β. The proliferation of RASF was measured by BrdU incorporation. Cell culture inserts were used to evaluate cell migration. For adhesion assays, RASF were seeded in culture plates. Then, plates were extensively shaken and adherent RASF quantified. Cell viability, cytotoxicity and apoptosis were measured using the ApoTox-Glo™ Triplex and the CellTox™ Green Cytotoxicity Assay.
Results: Tofacitinib and baricitinib decreased the IL-6 release of RASF stimulated with oncostatin M. JAK inhibition attenuated the IL-6 release of IL-1β activated and with soluble IL-6 receptor treated RASF. In contrast, only peficitinib and filgotinib decreased the IL-6 release of RASF activated with IL-1β. Peficitinib decreased also the MMP-3, CXCL8, and CXCL1 release at 5 μM. Moreover, peficitinib was the only JAK inhibitor suppressing proliferation of activated RASF at 1 μM. Peficitinib further decreased the migration of RASF without being cytotoxic or pro-apoptotic and without altering cell adhesion.
Conclusions: JAK inhibitors effectively suppress the inflammatory response induced by oncostatin M and by transsignaling of IL-6 in RASF. Only peficitinib modulated the IL-1β-induced response of RASF and their proliferation in vitro at concentrations close to reported Cmax values of well tolerated doses in vivo. In contrast to filgotinib, peficitinib also highly suppressed RASF migration showing the potential of peficitinib to target RASF.
A taxonomic revision of Desplatsia Bocq. (Malvaceae s. lat. Juss., subfamily Grewioideae Hochr., tribe Grewieae Endl.) based on about 800 herbarium specimens is presented. Desplatsia is a genus of trees and shrubs found in tropical West and Central Africa and is characterized by subulately divided stipules, the absence of an androgynophore, stamens that are fused to a tube at the base, and large and distinctive fruits that are dispersed by elephants. Four species are recognized (D. subericarpa Bocq., D. chrysochlamys (Mildbr. & Burret) Mildbr. & Burret, D. dewevrei (De Wild. & T.Durand) Burret and D. mildbraedii Burret) and 12 species names are placed into synonymy, two of which have been put into synonymy for the first time: D. floribunda Burret syn. nov. and D. trillesiana (Pierre ex De Wild.) Pierre ex A.Chev. syn. nov. All four species are widely distributed and their conservation status is assessed as Least Concern (LC). A key to the species, full species descriptions, illustrations, a specimen citation list and distribution maps are provided.
The TP53-induced glycolysis and apoptosis regulator (TIGAR) has been shown to decrease glycolysis, to activate the pentose phosphate pathway, and to provide protection against oxidative damage. Hypoxic regions are considered characteristic of glioblastoma and linked with resistance to current treatment strategies. Here, we established that LNT-229 glioma cell lines stably expressed shRNA constructs targeting TIGAR, and exposed them to hypoxia, irradiation and temozolomide. The disruption of TIGAR enhanced levels of reactive oxygen species and cell death under hypoxic conditions, as well as the effectiveness of irradiation and temozolomide. In addition, TIGAR was upregulated by HIF-1α. As a component of a complex network, TIGAR contributes to the metabolic adjustments that arise from either spontaneous or therapy-induced changes in tumor microenvironment.
As the prognosis of invasive aspergillosis remains unacceptably poor in patients undergoing hematopoietic stem cell transplantation (HSCT), there is a growing interest in the adoptive transfer of antifungal effector cells, such as Natural Killer (NK) cells. Because immunosuppressive agents are required in most HSCT recipients, knowledge of the impact of these compounds on the antifungal activity of NK cells is a prerequisite for clinical trials. We, therefore, assessed the effect of methylprednisolone (mPRED), cyclosporin A (CsA) and mycophenolic acid (MPA) at different concentrations on proliferation, apoptosis/necrosis, and the direct and indirect anti-Aspergillus activity of human NK cells. Methylprednisolone decreased proliferation and increased apoptosis of NK cells in a significant manner. After seven days, a reduction of viable NK cells was seen for all three immunosuppressants, which was significant for MPA only. Cyclosporin A significantly inhibited the direct hyphal damage by NK cells in a dose-dependent manner. None of the immunosuppressive compounds had a major impact on the measured levels of interferon-γ, granulocyte-macrophage colony-stimulating factor and RANTES (regulated on activation, normal T cell expressed and secreted; CCL5). Our data demonstrate that commonly used immunosuppressive compounds have distinct effects on proliferation, viability and antifungal activity of human NK cells, which should be considered in designing studies on the use of NK cells for adoptive antifungal immunotherapy.
In today’s "new world of work," knowledge workers are often given considerable flexibility regarding where and when to work (i.e., time-spatial flexibility) and this has become a popular approach to redesigning work. Whilst the adoption of such practices is mainly considered a top-down approach to work design, we argue that successful utilization of time-spatial flexibility requires proactivity on the part of the employee in the form of time-spatial job crafting. Previous research has demonstrated that time-spatial flexibility can have both positive and negative effects on well-being, performance, and work-life balance; yet remains mute about the underlying reasons for this and how employees can handle the given flexibility. Drawing on research from work design, we posit that in order for employees to stay well and productive in this context, they need to engage in time-spatial job crafting (i.e., a context-specific form of job crafting that entails reflection on time and place), which can be considered a future work skill. We propose a theoretical model of time-spatial job crafting in which we discuss its components, shed light on its antecedents, and explain how time-spatial job crafting is related to positive work outcomes through a time/spatial-demands fit.
Antagonistic and mutualistic species interactions provide important ecosystem functions affecting plant population dynamics and distribution. Many of these functions are important for the regeneration of plants, either by limiting or facilitating successful transition between life stages. Interactions can occur across the whole geographical range of a species and thereby encompass different environmental gradients, such as changes in temperature or water availability. Understanding the joint effects of species interactions and environmental factors on the regeneration of plants is key for understanding plant population dynamics under global change and could provide important recommendations for managing and conservation efforts.
My thesis aimed at advancing the knowledge of how species interactions depend on environmental conditions and jointly affect plant recruitment along the elevational distribution of plants. This thesis includes three chapters in which I studied the effects of animal seed deposition, seed predation, mycorrhizal and pathogenic fungi occurrences as well as abiotic and biotic environmental factors on the recruitment of Swiss stone pine (Pinus cembra). I conducted fieldwork in the Swiss Alps across the entire elevational distribution of the pine (1850 – 2250 m a.s.l). Over a period of three years, I recorded animal seed deposition by spotted nutcrackers (Nucifraga caryocatactes) and conducted seed translocation experiments. Further, I assessed fungal communities using DNA metabarcoding. I measured abiotic environmental factors such as temperature, water and light availability, pH, as well as biotic environmental factors such as distance to conspecific adults and ground vegetation cover. In my thesis, I used a broad range of community ecology approaches, from seed dispersal ecology to experimental plant ecology and microbial ecology.
First, I investigated the effects of environmental factors on four recruitment processes (i.e. seed deposition, seed predation, seed germination, seedling survival) of Swiss stone pine. Further, I aimed at identifying the most important recruitment processes potentially limiting pine regeneration across its elevational range. To investigate pine recruitment, I firstly tested how seed deposition, seed predation, seed germination and seedling survival were affected by the microhabitat characteristics ultimately determining where a seed arrives in the environment (i.e. canopy cover & ground vegetation cover). Secondly, I applied a sensitivity analysis to investigate which of the four recruitment processes poses limitation to the pines’ regeneration across its range. My results reveal that the importance of particular recruitment processes varies along the pines’ elevational range. I found that at the lower range margin and the distribution centre seed germination and seedling survival were the main limiting factors, whereas animal-mediated seed dispersal became especially important at the upper range margin. My study contributes to the field with a new approach for disentangling the relative importance of recruitment processes across environmental gradients and thereby could help to project how plant recruitment might respond to future changes in environmental conditions.
The second aim of my study was to investigate how abiotic and biotic environmental factors affect the occurrence of Swiss stone pine-associated pathogenic and mutualistic fungi by combining field measurements of environmental factors with a DNA metabarcoding approach. I identified potentially important fungal interaction partners of the pine and determined drivers shaping their occurrences. My results reveal that generalist fungi were not affected by abiotic and biotic environmental factors. However, specialist pathogens showed patterns according to the Janzen-Connell framework (i.e. accumulation of pathogen close to adult plants). Interestingly, I found evidence for an “inverse” Janzen-Connell effect, i.e. high abundance of a specialist mutualist close to adult plants, potentially mitigating effects of soil pathogens close to parent trees. Further, I found that pine-associated fungi are distributed widely within and beyond the range of their host plant, adding knowledge on how mutualisms and antagonisms might be affected when plants move their distributional range upwards.
Finally, I investigated how known and unknown plant-associated fungi affect the regeneration of Swiss stone pine in an environmental context. My results suggest that seedling establishment was most strongly affected by abiotic environmental factors, such as light availability and maximum summer temperature. Further, the results indicate that seedling survival was affected by biotic environmental factors, i.e. fungal agents, with high abundances of a known fungal pathogen co-occurring with low seedling survival rates. My results also reveal that known mycorrhizal partners as well as a large number of unknown fungal operational taxonomic units (OTUs) were associated with the survival of seedlings. My findings highlight the importance of plant-fungal interactions for plant recruitment and offer a feasible approach for the identification of hidden plant-fungal associations in highly complex DNA metabarcoding datasets. This approach offers a valuable tool for investigating plant-microbe interactions, ultimately helping to understand plant population dynamics.
My dissertation adds to a deeper understanding on the linkage between plant regeneration and species interactions, especially on how plant-animal and plant-fungal interactions in concert with environmental factors shape plant recruitment. My study reveals the importance of animal-mediated seed dispersal and fungal pathogens in plant recruitment with consequences for potential range shifts of plant species. My thesis has important implications for conservation and management efforts by informing on key species interactions under environmental change.
Understanding global biodiversity patterns is one of the main objectives of ecology. Spatial variation in species richness can be explained by several environmental factors. The relationships between species richness and environmental factors have been associated with latitudinal, longitudinal and elevational gradients. The number of species is determined by birth, death and migration rates of species in a given area. These rates are affected by abiotic and biotic factors acting at local and regional scales. Climatic seasonal variation may also influence biodiversity, directly through physiological limitations and indirectly through biotic interactions, vegetation structure and food availability. Climate and land use change are the main factors for landscape simplification and biotic homogenization. Thus, the study of community patterns across environmental gradients may help to predict the effect of projected environmental change.
I investigated how abiotic and biotic factors influence different facets of bird diversity across an elevational gradient. My study was conducted along an elevational gradient spanning 2000 m within and around Podocarpus National Park and San Francisco reserve on the southeastern slope of the Andes in Ecuador. The climate is humid tropical montane with a bimodal rain regime. The region is characterized by evergreen premontane forest at low elevations, evergreen lower montane forest at mid elevations and upper montane forest at high elevations. The elevational gradient has natural continuous forests within the protected reserves and fragmented forests surrounding the reserves in a matrix of cattle pastures. To monitor bird diversity, I placed nine 20-m radius point counts within 18 one-hectare plots, in continuous and fragmented forest at 1000, 2000 and 3000 m a.s.l. I recorded and identified all birds for 10 minutes within each point count. Bird communities were sampled eight times per plot, in the most humid season and in the least humid season of 2014 and 2015. To estimate flower and fruit availability, I recorded all plants with open flowers and ripe fruits within each point count. To obtain the relative invertebrate availability, I assessed understory invertebrate fresh biomass using a standardized sweep-netting design along 100-metre borders of each plot. Vertical vegetation heterogeneity was estimated at eight layers above the ground within each point count. Temperature for each plot was obtained using an air temperature regionalization tool and precipitation through remote sensing techniques and meteorological data.
In the first chapter of this thesis, I explored the effects of elevation, climate and vegetation structure on overall bird communities as well as on frugivorous and insectivorous birds. I found that elevation was mostly indirectly associated with bird diversity, jointly mediated via temperature, precipitation and vegetation structure. Additionally, elevation was directly and positively associated with both the overall bird community and with insectivores, but not with frugivores. My findings indicate a reduction of bird diversity due to climatic factors and vegetation structure with increasing elevation. However, the direct, positive effect of elevation suggests that bird diversity was higher than expected towards high elevations, probably due to spatial, biotic and evolutionary settings.
In the second chapter, I analysed the influence of climate and resource availability on temporal variation of bird communities. I found a higher bird diversity in the least humid season than in the most humid season. The seasonality of the bird communities was mainly driven by temperature and precipitation. While temperature had a significant positive effect at high elevations, precipitation had a significant negative effect at low elevations. Resource availability had no significant effect. My findings suggest that the temporal fluctuations in bird communities likely occur due to climate
constraints rather than due to resource limitations.
In the third chapter, I studied the effect of forest fragmentation on taxonomic and functional bird diversity. I found that taxonomic diversity was higher in fragmented compared to continuous forests, while functional diversity was negatively affected by fragmentation, but only at low elevations. The increase of taxonomic diversity in disturbed habitats suggests an increase of habitat generalists, which may compensate the loss of forest specialists. My findings suggest that taxonomic diversity can be uncoupled from functional diversity in diverse communities at low elevations.
My results show the effects of environmental factors on the spatio-temporal patterns of bird communities and the potentially uncoupled responses of taxonomic and functional diversity to forest fragmentation. My findings highlight that bird communities respond differently to abiotic and biotic factors across elevational gradients. Overall, my study helps to better understand the mechanisms that drive species communities in response to complex environmental conditions, which could be an essential contribution for the conservation of bird communities in the tropical Andes.
Since ethnic stereotyping gained new political virulence in the current ethnopopulist climate, the three-day conference, organized by five researchers from the Department of Comparative Literature at the University of Vienna, aimed to promote academic discussion on new perspectives on imagology. In twenty-six presentations, international scholars shared their findings on strategies of Othering within a transdisciplinary framework of different theoretical approaches from postcolonial theory to gender studies, from intermediality to musicology, in order to fathom the boundaries of imagological research today. An additional poster session gave students the opportunity to present their research on hetero- and auto-stereotypes within literature.
The recognition of pharmacological substances, compounds and proteins is an essential preliminary work for the recognition of relations between chemicals and other biomedically relevant units. In this paper, we describe an approach to Task 1 of the PharmaCoNER Challenge, which involves the recognition of mentions of chemicals and drugs in Spanish medical texts. We train a state-of-the-art BiLSTM-CRF sequence tagger with stacked Pooled Contextualized Embeddings, word and sub-word embeddings using the open-source framework FLAIR. We present a new corpus composed of articles and papers from Spanish health science journals, termed the Spanish Health Corpus, and use it to train domain-specific embeddings which we incorporate in our model training. We achieve a result of 89.76% F1-score using pre-trained embeddings and are able to improve these results to 90.52% F1-score using specialized embeddings.
Motivation: Complexome profiling combines native gel electrophoresis with mass spectrometry to obtain the inventory, composition and abundance of multiprotein assemblies in an organelle. Applying complexome profiling to determine the effect of a mutation on protein complexes requires separating technical and biological variations from the variations caused by that mutation.
Results: We have developed the COmplexome Profiling ALignment (COPAL) tool that aligns multiple complexome profiles with each other. It includes the abundance profiles of all proteins on two gels, using a multi-dimensional implementation of the dynamic time warping algorithm to align the gels. Subsequent progressive alignment allows us to align multiple profiles with each other. We tested COPAL on complexome profiles from control mitochondria and from Barth syndrome (BTHS) mitochondria, which have a mutation in tafazzin gene that is involved in remodeling the inner mitochondrial membrane phospholipid cardiolipin. By comparing the variation between BTHS mitochondria and controls with the variation among either, we assessed the effects of BTHS on the abundance profiles of individual proteins. Combining those profiles with gene set enrichment analysis allows detecting significantly affected protein complexes. Most of the significantly affected protein complexes are located in the inner mitochondrial membrane (mitochondrial contact site and cristae organizing system, prohibitins), or are attached to it (the large ribosomal subunit).
Availability and implementation: COPAL is written in python and is available from http://github.com/cmbi/copal.
With an increased understanding of the tumor biology of squamous cell carcinoma of the head and neck (SCCHN), targeted therapies have found their way into the clinical treatment routines against this entity. Nevertheless, to date platinum-based cytostatic agents remain the first line choice and targeting the epidermal growth factor-receptor (EGFR) with combined cetuximab and radiation therapy remains the only targeted therapy approved in the curative setting. Investigation of immune checkpoint inhibitors (ICI), such as antibodies targeting programmed cell death protein 1 (PD-1) and its ligand PD-L1, resulted in a change of paradigms in oncology and in the first approval of new drugs for treating SCCHN. Nivolumab and pembrolizumab, two anti-PD-1 antibodies, were the first agents shown to improve overall survival for patients with metastatic/recurrent tumors in recent years. Currently, several clinical trials investigate the role of ICI in different therapeutic settings. A robust set of biomarkers will be an inevitable tool for future individualized treatment approaches including radiation dose de-escalation and escalation strategies. This review aims to summarize achieved goals, the current status and future perspectives regarding targeted therapies and ICI in the management of SCCHN.
The title co-crystal, 1,3,5,7-tetraazatricyclo[3.3.1.13,7]decane (HMTA, 1)–4-fluorophenol (4-FP) (1/1), C6H12N4·C6H5FO, shows an unusual asymmetric unit that comprises eight independent molecules (Z′′ = 8), four for each component, with four formula units per asymmetric unit (Z′ = 4). In the molecular packing, each HMTA molecule bridges one 4-FP molecule via an O−H···N hydrogen bond to form a two-molecule aggregate. Differences can be observed between the bond lengths and angles of the independent HMTA and 4-FP molecules and those of the molecules in the aggregate. The C−N bonds exhibit different bond lengths in the tetrahedral cage-like structure of the HMTA molecules, but the largest differences between the molecular aggregates are in the bond lengths in the 4-fluorophenol ring. In the crystal, the HMTA and 4-FP molecules form two hydrogen-bonded (O−H···N, C−H···F and C−H···O) dimers of HMTA and 4-FP molecules, A···D and B···C inversion dimers, which generate enlarged R88(34) ring motifs in both supramolecular structures. In both structures, the crystal packing also features additional C−H···F and C−H···O interactions. The A···D and B···C dimers are linked by additional C−H···F and C−H···O hydrogen bonds, forming columns along the a and b axes, respectively. The importance of the C−H···F interaction to the structure and crystal packing has been demonstrated.
Three experiments investigated the interpretation and production of pronouns in German. The first two experiments probed the preferred interpretation of a pronoun in contexts containing two potential antecedents by having participants complete a sentence fragment starting either with a personal pronoun or a d-pronoun. We systematically varied three properties of the potential antecedents: syntactic function, linear position, and topicality. The results confirm a subject preference for personal pronouns. The preferred interpretation of d-pronouns cannot be captured by any of the three factors alone. Although a d-pronoun preferentially refers to the non-topic in many cases, this preference can be overridden by the other two factors, linear position and syntactic function. In order to test whether interpretive preferences follow from production biases as proposed by the Bayesian theory of Kehler et al. (2008), a third experiment had participants freely produce a continuation sentence for the contexts of the first two experiments. The results show that personal pronouns are used more often to refer to a subject than to an object, recapitulating the subject preference found for interpretation and thereby confirming the account of Kehler et al. (2008). The interpretation results for the d-pronoun likewise follow from the corresponding production data.
Background: Reports of head and neck ultrasound examinations are frequently written by hand as free texts. Naturally, quality and structure of free text reports is variable, depending on the examiner’s individual level of experience. Aim of the present study was to compare the quality of free text reports (FTR) and structured reports (SR) of head and neck ultrasound examinations.
Methods: Both standard FTRs and SRs of head and neck ultrasound examinations of 43 patients were acquired by nine independent examiners with comparable levels of experience. A template for structured reporting of head and neck ultrasound examinations was created using a web-based approach. FTRs and SRs were evaluated with regard to overall quality, completeness, required time to completion, and readability by four independent raters with different specializations (Paired Wilcoxon test, 95% CI) and inter-rater reliability was assessed (Fleiss’ kappa). A questionnaire was used to compare FTRs vs. SRs with respect to user satisfaction (Mann-Whitney U test, 95% CI).
Results: By comparison, completeness scores of SRs were significantly higher than FTRs’ completeness scores (94.4% vs. 45.6%, p < 0.001), and pathologies were described in more detail (91.1% vs. 54.5%, p < 0.001). Readability was significantly higher in all SRs when compared to FTRs (100% vs. 47.1%, p < 0.001). The mean time to complete a report, however, was significantly higher in SRs (176.5 vs. 107.3 s, p < 0.001). SRs achieved significantly higher user satisfaction ratings (VAS 8.87 vs. 1.41, p < 0.001) and a very high inter-rater reliability (Fleiss’ kappa 0.92).
Conclusions: As compared to FTRs, SRs of head and neck ultrasound examinations are more comprehensive and easier to understand. On the balance, the additional time needed for completing a SR is negligible. Also, SRs yield high inter-rater reliability and may be used for high-quality scientific data analyses.
State-of-the-art climate models contain, to a significant degree, empirical components. In particular, subgrid-scale (SGS) parameterizations are usually highly tuned against observations or high-resolution model data. While this enables the models to minimize the error during hindcasts, it is not guaranteed that it yields a benefit for climate projections because of climate change. In this thesis the Fluctuation-Dissipation theorem (FDT) is used to update the statistics of the system in the presence of an external forcing. If the empirical parameters are tuned objectively to the data (i.e., they depend on the statistics of the data), then they might be updated with the FDT. This ansatz is tested within a framework of a semi-empirical model (SEM) based on the leading variance patterns of a quasigeostrophic three-layer model (QG3LM) and supplemented by a purely data-driven parameterization. We show that the FDT is able to successfully update the tuning parameters of the data-driven SGS closure, resulting in a systematic improvement in model performance in comparison to an untreated SEM. Ideally, SGS parameterizations should contain little to no tuning parameters. Thus, complementary to the FDT approach we investigate a stochastic SGS closure constrained by first principles that is calculated using the stochastic mode reduction (SMR). The SMR allows for an analytic derivation of the SGS closure from the model equations while requiring only minimal tuning. We successfully apply the SMR to the QG3LM and construct the reduced stochastic model (RSM). Furthermore, we show that the RSM is more robust against an external forcing than the SEM. Additionally, we find that, under appropriate conditions, the FDT is able to update the empirical parts of the RSM. Yet, only for the response in mean streamfunction the RSM provides useful results, while the response in covariance of the streamfunction is incorrect for most cases. Nevertheless, we obtain a remarkably accurate response in both moments for the RSM in an idealized setting. In combination with the results of the FDT study this indicates that the considered RSM is too low dimensional and encourages us to investigate the response of larger RSMs in the future.
The research focuses on magic - the practice of performing tricks and illusions on stage aiming at entertaining the audience. In the late XIX-early XX century magic achieved an outstanding social recognition and became an important artistic phenomenon. This study aims to analyze the work of the most prominent magicians of the late XIX-early XX centuries and to define the historical role of magicians in the history of culture and art.
Using methods of art history and cultural studies, I analyze the autobiographies of magicians, the literature on magic published during the period in question, and contemporary press. I approach the history of magic from three different perspectives: magic as a branch of show business, magic as a cultural phenomenon, and magic as a type of performing art. It allowed me to create a detailed account of magic as a social, cultural and artistic phenomenon.
I argue that magic became a highly influential cultural phenomenon of the late 19th- early 20th centuries in Great Britain that represented the idea of real magic on stage. Magic shows reflected a complex relationship between rational and magical thinking that existed in society and produced narratives about science and the supernatural.
Moreover, very few studies have focused on the question of defining magic as an art form. In the thesis, I analyzed the theoretical works on magic written by magicians and developed a framework for further research on magic from the perspective of the history of art.
Different tissue engineering techniques are used to support rapid vascularisation. A novel technique is the use of platelet-rich fibrin (PRF), an autologous source of growth factors. This study was the first to investigate the influence of PRF matrices, isolated following different centrifugation protocols, on human dermal vascular endothelial cells (ECs) in mono-culture and co-culture with human primary fibroblasts (HFs) as an in vitro model for tissue regeneration. Focus was placed on vascular structure formation and growth factor release. HFs and ECs were cultivated with PRF prepared using a high (710 ×g) or low (44 ×g) relative centrifugation force (RCF) over 14 d. Immunofluorescence staining and immunohistochemistry were used to evaluate the microvascular formation. Cell culture supernatants were collected for evaluation of growth factor release. The results showed a PRF-mediated effect on the induction of angiogenesis in ECs. Microvessel-like structure formation was promoted when ECs were combined with low-RCF PRF as compared to high-RCF PRF or control group. The percentage of vascular lumen area was significantly higher in low-RCF PRF, especially at day 7, which coincided with statistically significantly higher growth factor [vascular endothelial factor (VEGF), transforming growth factor β1 (TGF-β1) and platelet derived growth factor (PDGF)] concentration measured in low-RCF PRF as compared to high-RCF PRF or control group. In conclusion, reducing the RCF according to the low-speed centrifugation concept (LSCC) resulted in increased growth factor release and angiogenic structure formation with EC mono-culture, suggesting that PRF may be a highly beneficial therapeutic tool for tissue engineering applications.
Platelet-rich fibrin (PRF) is a blood concentrate derived from venous blood that is processed without anticoagulants by a one-step centrifugation process. This three-dimensional scaffold contains inflammatory cells and plasma proteins entrapped in a fibrin matrix. Liquid-PRF was developed based on the previously described low-speed centrifuge concept (LSCC), which allowed the introduction of a liquid-PRF formulation of fibrinogen and thrombin prior to its conversion to fibrin. Liquid-PRF was introduced to meet the clinical demand for combination with biomaterials in a clinically applicable and easy-to-use way. The aim of the present study was to evaluate, ex vivo, the interaction of the liquid-PRF constituents with five different collagen biomaterials by histological analyses. The results first demonstrated that large variability existed between the biomaterials investigated. Liquid-PRF was able to completely invade Mucograft® (MG; Geistlich Biomaterials, Wolhusen, Switzerland) and to partly invade Bio-Gide® (BG; Geistlich Biomaterials, Wolhusen, Switzerland) and Mucoderm® (MD; Botiss Biomaterials, Berlin, Germany), and Collprotect® (CP; Botiss Biomaterials, Berlin, Germany) showed only a superficial interaction. The BEGO® collagen membrane (BCM; BEGO Implant Systems) appeared to be completely free of liquid-PRF. These results were confirmed by the different cellular penetration and liquid-PRF absorption coefficient (PAC) values of the evaluated membranes. The present study demonstrates a system for loading biomaterials with a complex autologous cell system (liquid-PRF) in a relatively short period of time and in a clinically relevant manner. The combination of biomaterials with liquid-PRF may be clinically utilized to enhance the bioactivity of collagen-based biomaterials and may act as a biomaterial-based growth factor delivery system.
Attractiveness ratings for musicians and non-musicians: An evolutionary-psychology perspective
(2019)
From an evolutionary perspective, musical behavior such as playing an instrument can be considered as part of an individual’s courting behavior. Playing a musical instrument or singing might fulfill a function similar to that of a bird’s colored feathers: attracting attention. Therefore, musicians may be rated as more attractive than non-musicians. In an online survey, 137 volunteers (95 female) with ages ranging from 16 to 39 years rated the attractiveness of fictitious persons of the opposite sex described in short verbal profiles. These profiles differed with respect to whether the described person made music or not. Additionally, the musicians’ profiles varied with regard to whether the described person played music or sang in public or in private only. Results show that musicians’ profiles were not generally rated as more attractive than non-musicians’, but attractiveness did vary according to setting: private musicians were rated as most attractive, followed by non-musicians and public musicians. Furthermore, results indicate that participants who played a musical instrument or sang themselves gave higher ratings to profiles of musicians. But for participants who do not make music themselves, higher attractiveness ratings for musicians playing instruments or sing in private settings were found. These results indicate that the impression of sharing a common interest (making music) and furthermore making music in private instrumental settings seems to make people attractive to other people. No additional support for the sexual selection hypotheses for the evolution of music was provided by the current results. The musical status of the rater affected his or her judgements, with musicians rating other people as more attractive if they share the common interest in making music. Not the display of being a musician seems to be critical for attractiveness ratings but the perceived or imagined similarity by the rater created by information on musicality, fostering the theoretical significance of the communication aspect of music.
Chimeric antigen receptor (CAR) T cells are in prime focus of current research in cancer immunotherapy. Facilitating CAR T cell generation is among the top goals. We have recently demonstrated direct in vivo generation of human CD19-CAR T cells by targeting CD8+ cells using lentiviral vectors (LVs). The anti-tumor potency of in vivo generated CAR T cells was assessed in human PBMC-transplanted NSG mice carrying i.v. injected CD19+ Nalm-6 tumor cells. A single injection of CD8-targeted LV delivering CD19-CAR was sufficient to completely eliminate the tumor cells from bone marrow and spleen, whereas control animals contained high levels of CD19+ cells. Tumor elimination was due to in vivo generated CAR+ cells. Notably, these were not only composed of T lymphocytes but also included CAR+ natural killer cells (NK and NKT). This is the first demonstration of tumor elimination by in vivo generated human CAR T cells.
Background: Network science provides powerful access to essential organizational principles of the brain. The aim of this study was to investigate longitudinal evolution of gray matter networks in early relapsing–remitting MS (RRMS) compared with healthy controls (HCs) and contrast network dynamics with conventional atrophy measurements.
Methods: For our longitudinal study, we investigated structural cortical networks over 1 year derived from 3T MRI in 203 individuals (92 early RRMS patients with mean disease duration of 12.1 ± 14.5 months and 101 HCs). Brain networks were computed based on cortical thickness inter-regional correlations and fed into graph theoretical analysis. Network connectivity measures (modularity, clustering coefficient, local efficiency, and transitivity) were compared between patients and HCs, and between patients with and without disease activity. Moreover, we calculated longitudinal brain volume changes and cortical atrophy patterns.
Results: Our analyses revealed strengthening of local network properties shown by increased modularity, clustering coefficient, local efficiency, and transitivity over time. These network dynamics were not detectable in the cortex of HCs over the same period and occurred independently of patients’ disease activity. Most notably, the described network reorganization was evident beyond detectable atrophy as characterized by conventional morphometric methods.
Conclusion: In conclusion, our findings provide evidence for gray matter network reorganization subsequent to clinical disease manifestation in patients with early RRMS. An adaptive cortical response with increased local network characteristics favoring network segregation could play a primordial role for maintaining brain function in response to neuroinflammation.
Global response of diacylglycerol kinase towards substrate binding observed by 2D and 3D MAS NMR
(2019)
Escherichia coli diacylglycerol kinase (DGK) is an integral membrane protein, which catalyses the ATP-dependent phosphorylation of diacylglycerol (DAG) to phosphatic acid (PA). It is a unique trimeric enzyme, which does not share sequence homology with typical kinases. It exhibits a notable complexity in structure and function despite of its small size. Here, chemical shift assignment of wild-type DGK within lipid bilayers was carried out based on 3D MAS NMR, utilizing manual and automatic analysis protocols. Upon nucleotide binding, extensive chemical shift perturbations could be observed. These data provide evidence for a symmetric DGK trimer with all of its three active sites concurrently occupied. Additionally, we could detect that the nucleotide substrate induces a substantial conformational change, most likely directing DGK into its catalytic active form. Furthermore, functionally relevant interprotomer interactions are identified by DNP-enhanced MAS NMR in combination with site-directed mutagenesis and functional assays.
Transport mechanism of a multidrug resistance protein investigated by pulsed EPR spectroscopy
(2019)
In human several diseases result from malfunctions of ATP-binding cassette (ABC) systems, which form one of the largest transport system superfamily. Many ABC exporters contain asymmetric nucleotide-binding sites (NBSs) and some of them are inhibited by the transported substrate.1 For the active transport of diverse chemically substrates across biological membranes, ABC transport complexes use the energy of ATP binding and subsequent hydrolysis. In this thesis, the heterodimeric ABC exporter TmrAB2,3 from Thermus thermophilus, a functional homolog of the human antigen translocation complex TAP, was investigated by using pulsed electron-electron double resonance (PELDOR/DEER) spectroscopy. In the presence of ATP, TmrAB exists in an equilibrium between inward- and outward-facing conformations. This equilibrium can be modulated by changing the ATP concentration, showing asymmetric behaviour in the open-to-close equilibrium between the consensus and the degenerate NBSs. At the degenerate NBS the closed conformation is more preferred and closure of one of the NBSs is sufficient to open the periplasmic gate at the transmembrane domain (TMD).3 By determining the temperature dependence of this conformational equilibrium, the thermodynamics of the energy coupling during ATP-induced conformational changes in TmrAB were investigated. The results demonstrate that ATP-binding alone drives the global conformational switching to the outward-facing state and allows the determination of the entropy and enthalpy changes for this step. With this knowledge, the Gibbs free energy of this ATP induced transition was calculated. Furthermore, an excess of substrate, meaning trans-inhibition of the transporter is resulting mechanistically in a reverse transition from the outward-facing state to an occluded conformation predominantly.3 This work unravels the central role of the reversible conformational equilibrium in the function and regulation of an ABC exporter. For the first time it is shown that the conformational thermodynamics of a large membrane protein complex can be investigated. The presented experiments give new possibilities to investigate other related medically important transporters with asymmetric NBSs or other similar protein complexes.
hallmark of ageing is the redistribution of body fat. Particularly, subcutaneous fat decreases paralleled by a decrease of skin collagen I are typical for age-related skin atrophy. In this paper, we hypothesize that collagen I may be a relevant molecule stimulating the differentiation of adipose-derived stem cells (ASCs) into adipocytes augmenting subcutaneous fat. In this context lipogenesis, adiponectin, and collagen I receptor expression were determined. Freshly isolated ASCs were characterized by stemness-associated surface markers by FACS analysis and then transdifferentiated into adipocytes by specific medium supplements. Lipogenesis was evaluated using Nile Red staining and documented by fluorescence microscopy or quantitatively measured by using a multiwell spectrofluorometer. Expression of adiponectin was measured by real-time RT-PCR and in cell-free supernatants by ELISA, and expression of collagen I receptors was observed by western blot analysis. It was found that supports coated with collagen I promote cell adhesion and lipogenesis of ASCs. Interestingly, a reverse correlation to adiponectin expression was observed. Moreover, we found upregulation of the collagen receptor, discoidin domain-containing receptor 2; receptors of the integrin family were absent or downregulated. These findings indicate that collagen I is able to modulate lipogenesis and adiponectin expression and therefore may contribute to metabolic dysfunctions associated with ageing.
Amorphous formulation technologies to improve oral absorption of poorly soluble active pharmaceutical ingredients (APIs) have become increasingly prevalent. Currently, polymer-based amorphous formulations manufactured by spray drying, hot melt extrusion (HME), or co-precipitation are most common. However, these technologies have challenges in terms of the successful stabilization of poor glass former compounds in the amorphous form. An alternative approach is mesoporous silica, which stabilizes APIs in non-crystalline form via molecular adsorption inside nano-scale pores. In line with these considerations, two poor glass formers, haloperidol and carbamazepine, were formulated as polymer-based solid dispersion via HME and with mesoporous silica, and their stability was compared under accelerated conditions. Changes were monitored over three months with respect to solid-state form and dissolution. The results were supported by solid-state nuclear magnetic resonance spectroscopy (SS-NMR) and scanning electron microscopy (SEM). It was demonstrated that mesoporous silica was more successful than HME in the stabilization of the selected poor glass formers. While both drugs remained non-crystalline during the study using mesoporous silica, polymer-based HME formulations showed recrystallization after one week. Thus, mesoporous silica represents an attractive technology to extend the formulation toolbox to poorly soluble poor glass formers.
The IPS e.max system by Ivoclar Vivadent, offering a variety of products and indications, is widely used for all-ceramic restorations. We analyzed the clinical track record of these products in daily clinical practice, associating their restorative survival rate with various parameters to define recommendations for long-term stability. A total of 1058 full-coverage crowns and fixed partial dentures (FPDs) were evaluated retrospectively over up to 66.48 (37.05 ± 18.4) months. All were made of IPS e.max Press, IPS e.max CAD, IPS e.max Ceram or IPS e.max ZirPress and had been delivered by a private dental practice within three years. Uses not recommended by the manufacturer were also deliberately included. The five-year cumulative survival was 94.22% (i.e., 94.69% or 90.58% for glass-ceramic crowns or FDPs and 100% or 90.06% for zirconia-based crowns or FDPs). Significantly superior outcomes emerged for conventional vs. adhesive cementation and for vital vs. non-vital abutment teeth, but not for recommended vs. non-recommended uses. Caution is required in restoring non-vital teeth, but the spectrum of recommended uses should generally be reconsidered and expanded, given our finding of high survival and success rates for IPS e.max ceramics, even for uses not currently recommended by the manufacturer.
Brain metastases are the most common intracranial tumor in adults and are associated with poor patient prognosis and median survival of only a few months. Treatment options for brain metastasis patients remain limited and largely depend on surgical resection, radio- and/or chemotherapy. The development and pre-clinical testing of novel therapeutic strategies require reliable experimental models and diagnostic tools that closely mimic technologies that are used in the clinic and reflect histopathological and biochemical changes that distinguish tumor progression from therapeutic response. In this study, we sought to test the applicability of magnetic resonance (MR) spectroscopy in combination with MR imaging to closely monitor therapeutic efficacy in a breast-to-brain metastasis model. Given the importance of radiotherapy as the standard of care for the majority of brain metastases patients, we chose to monitor the post-irradiation response by magnetic resonance spectroscopy (MRS) in combination with MR imaging (MRI) using a 7 Tesla small animal scanner. Radiation was applied as whole brain radiotherapy (WBRT) using the image-guided Small Animal Radiation Research Platform (SARRP). Here we describe alterations in different metabolites, including creatine and N-acetylaspartate, that are characteristic for brain metastases progression and lactate, which indicates hypoxia, while choline levels remained stable. Radiotherapy resulted in normalization of metabolite levels indicating tumor stasis or regression in response to treatment. Our data indicate that the use of MR spectroscopy in addition to MRI represents a valuable tool to closely monitor not only volumetrical but also metabolic changes during tumor progression and to evaluate therapeutic efficacy of intervention strategies. Adapting the analytical technology in brain metastasis models to those used in clinical settings will increase the translational significance of experimental evaluation and thus contribute to the advancement of pre-clinical assessment of novel therapeutic strategies to improve treatment options for brain metastases patients.
Myogenic vasoconstriction is an autoregulatory function of small arteries. Recently, G-protein-coupled receptors have been involved in myogenic vasoconstriction, but the downstream signalling mechanisms and the in-vivo-function of this myogenic autoregulation are poorly understood. Here, we show that small arteries from mice with smooth muscle-specific loss of G12/G13 or the Rho guanine nucleotide exchange factor ARHGEF12 have lost myogenic vasoconstriction. This defect was accompanied by loss of RhoA activation, while vessels showed normal increases in intracellular [Ca2+]. In the absence of myogenic vasoconstriction, perfusion of peripheral organs was increased, systemic vascular resistance was reduced and cardiac output and left ventricular mass were increased. In addition, animals with defective myogenic vasoconstriction showed aggravated hypotension in response to endotoxin. We conclude that G12/G13- and Rho-mediated signaling plays a key role in myogenic vasoconstriction and that myogenic tone is required to maintain local and systemic vascular resistance under physiological and pathological condition.
Objectives: To investigate the prevalence of depressive symptoms in rheumatoid arthritis (RA) patients using two previously validated questionnaires in a large patient sample, and to evaluate depressive symptoms in the context of clinical characteristics (e.g. remission of disease) and patient-reported impact of disease.
Methods: In this cross-sectional study, the previously validated Patient Health Questionnaire (PHQ-9) and Beck-Depression Inventory II (BDI-II) were used to assess the extent of depressive symptoms in RA patients. Demographic background, RA disease activity score (DAS28), RA impact of disease (RAID) score, comorbidities, anti-rheumatic therapy and antidepressive treatment, were recorded. Cut-off values for depressive symptomatology were PHQ-9 ≥5 or BDI-II ≥14 for mild depressive symptoms or worse and PHQ-9 ≥ 10 or BDI-II ≥ 20 for moderate depressive symptoms or worse. Prevalence of depressive symptomatology was derived by frequency analysis while factors independently associated with depressive symptomatology were investigated by using multiple logistic regression analyses. Ethics committee approval was obtained, and all patients provided written informed consent before participation.
Results: In 1004 RA-patients (75.1% female, mean±SD age: 61.0±12.9 years, mean disease duration: 12.2±9.9 years, DAS28 (ESR): 2.5±1.2), the prevalence of depressive symptoms was 55.4% (mild or worse) and 22.8% (moderate or worse). Characteristics independently associated with depressive symptomatology were: age <60 years (OR = 1.78), RAID score >2 (OR = 10.54) and presence of chronic pain (OR = 3.25). Of patients classified as having depressive symptoms, only 11.7% were receiving anti-depressive therapy.
Conclusions: Mild and moderate depressive symptoms were common in RA patients according to validated tools. In routine clinical practice, screening for depression with corresponding follow-up procedures is as relevant as incorporating these results with patient-reported outcomes (e.g. symptom state), because the mere assessment of clinical disease activity does not sufficiently reflect the prevalence of depressive symptoms.
Clinical trial registration number: This study is registered in the Deutsches Register Klinischer Studien (DRKS00003231) and ClinicalTrials.gov (NCT02485483).
Elucidating the immune evasion mechanisms of borrelia mayonii, the causative agent of lyme disease
(2019)
Borrelia (B.) mayonii sp. nov. has recently been reported as a novel human pathogenic spirochete causing Lyme disease (LD) in North America. Previous data reveal a higher spirochaetemia in the blood compared to patients infected by LD spirochetes belonging to the B. burgdorferi sensu lato complex, suggesting that this novel genospecies must exploit strategies to overcome innate immunity, in particular complement. To elucidate the molecular mechanisms of immune evasion, we utilized various methodologies to phenotypically characterize B. mayonii and to identify determinants involved in the interaction with complement. Employing serum bactericidal assays, we demonstrated that B. mayonii resists complement-mediated killing. To further elucidate the role of the key regulators of the alternative pathway (AP), factor H (FH), and FH-like protein 1 (FHL-1) in immune evasion of B. mayonii, serum adsorption experiments were conducted. The data revealed that viable spirochetes recruit both regulators from human serum and FH retained its factor I-mediated C3b-inactivating activity when bound to the bacterial cells. In addition, two prominent FH-binding proteins of approximately 30 and 18 kDa were detected in B. mayonii strain MN14-1420. Bioinformatics identified a gene, exhibiting 60% identity at the DNA level to the cspA encoding gene of B. burgdorferi. Following PCR amplification, the gene product was produced as a His-tagged protein. The CspA-orthologous protein of B. mayonii interacted with FH and FHL-1, and both bound regulators promoted inactivation of C3b in the presence of factor I. Additionally, the CspA ortholog counteracted complement activation by inhibiting the alternative and terminal but not the classical and Lectin pathways, respectively. Increasing concentrations of CspA of B. mayonii also strongly affected C9 polymerization, terminating the formation of the membrane attack complex. To assess the role of CspA of B. mayonii in facilitating serum resistance, a gain-of-function strain was generated, harboring a shuttle vector allowing expression of the CspA encoding gene under its native promotor. Spirochetes producing the native protein on the cell surface overcame complement-mediated killing, indicating that CspA facilitates serum resistance of B. mayonii. In conclusion, here we describe the molecular mechanism utilized by B. mayonii to resists complement-mediated killing by capturing human immune regulators.
The continental expression of global cooling during the Miocene Climate Transition in Central Asia is poorly documented, as the tectonically active setting complicates the correlation of Neogene regional and global climatic developments. This study presents new geochemical data (CaSO4 content, carbonate δ13C and δ18O) from the endorheic alluvial‐lacustrine Aktau succession (Ili Basin, south‐east Kazakhstan) combined with findings from the previously published facies evolution. Time series analysis revealed long‐eccentricity forcing of the paleohydrology throughout the entire succession, split into several facies‐dependent segments. Orbital tuning, constrained by new laser ablation U‐Pb dates and a preexisting magnetostratigraphy, places the succession in a 5.0 Ma long interval in the middle to late Miocene (15.6 to 10.6 Ma). The long‐term water accumulation in the Ili Basin followed the timing of the Miocene Climate Transition, suggesting increased precipitation in the catchment area in response to climate cooling and stronger westerly winds. This was paced by minima of the 2.4 Ma eccentricity cycle, which favored the establishment of a discharge playa (~14.3 Ma) and a perennial lake (12.6 to 11.8 Ma). Furthermore, low obliquity amplitudes (nodes) caused a transient weakening of the westerlies at ~13.7 to 13.5 Ma and at ~12.7 Ma, resulting in negative hydrological budgets and salinization. Flooding of the windward Ili Basin coeval with aridification in the leeward basins suggests that the Tian Shan was a climate boundary already in the middle Miocene. Our results emphasize the impact of climate fluctuations on the westerlies' strength and thus on Central Asian hydrology.
Objectives: Large-scale clinical studies investigating associations between intestinal microbiota signatures and human diseases usually rely on stool samples. However, the timing of repeated stool sample collection cannot be predefined in longitudinal settings. Rectal swabs, being straightforward to obtain, have the potential to overcome this drawback. Therefore, we assessed the usability of rectal swabs for microbiome sampling in a cohort of hematological and oncological patients.
Study design: We used a pipeline for intestinal microbiota analysis from deep rectal swabs which was established and validated with test samples and negative controls. Consecutively, a cohort of patients from hematology and oncology wards was established and weekly deep rectal swabs taken during their admissions and re-admissions.
Results: Validation of our newly developed pipeline for intestinal microbiota analysis from rectal swabs revealed consistent and reproducible results. Over a period of nine months, 418 rectal swabs were collected longitudinally from 41 patients. Adherence to the intended sampling protocol was 97%. After DNA extraction, sequencing, read pre-processing and filtering of chimeric sequences, 405 of 418 samples (96.9%) were eligible for further analyses. Follow-up samples and those taken under current antibiotic exposure showed a significant decrease in alpha diversity as compared to baseline samples. Microbial domination occurred most frequently by Enterococcaceae (99 samples, 24.4%) on family level and Enterococcus (90 samples, 22.2%) on genus level. Furthermore, we noticed a high abundance of potential skin commensals in 99 samples (24.4%).
Summary: Deep rectal swabs were shown to be reliable for microbiome sampling and analysis, with practical advantages related to high sampling adherence, easy timing, transport and storage. The relatively high abundance of putative skin commensals in this patient cohort may be of potential interest and should be further investigated. Generally, previous findings on alpha diversity dynamics obtained from stool samples were confirmed.
Background: Previous research demonstrated atypical attention in children with attention‐deficit/hyperactivity disorder (ADHD) and autism spectrum disorder (ASD). Regarding visual orienting, findings suggest a differential impairment: Atypical orienting to relatively unexpected targets in ASD, and atypical processing of alerting cues in ADHD. The locus coeruleus‐norepinephrine (LC‐NE) system plays an important role in exploiting alerting cues to increase attention and task performance. The present study’s aim was to examine differential subcortical processes underlying visual orienting in ASD and ADHD with pupil dilation (PD) as index of LC activity.
Methods: Pupil dilation (PD) progression metrics during visual orienting were calculated for task‐evoked PD locked to cue, stimulus onset, and behavioral response. Group differences in PD and reaction time (RT) were compared between children with ASD without ADHD (ASD‐) (N = 18), ADHD without ASD (ADHD‐) (N = 28), both disorders (ASD + ADHD) (N = 14), and typically developing children (TD) (N = 31) using linear mixed models (LMM). To further explore the modulatory role of the LC‐NE system group differences in the effect of task‐evoked PD metrics on RT were examined exploratively.
Results: ASD (+ADHD) showed slower orienting responses to relatively unexpected spatial target stimuli as compared to TD, which was accompanied by higher PD amplitudes relative to ADHD− and TD. In ADHD−, shorter cue‐evoked PD latencies relative to ASD−, ASD + ADHD, and TD were found. Group differences in the effect of cue‐ and stimulus‐evoked PD amplitudes on RT were found in ASD− relative to TD.
Conclusions: Study findings provide new evidence for a specific role of the LC‐NE system in impaired reflexive orienting responses in ASD, and atypical visual processing of alerting cues in ADHD.
Constitutive Wnt activation upon loss of Adenoma polyposis coli (APC) acts as main driver of colorectal cancer (CRC). Targeting Wnt signaling has proven difficult because the pathway is crucial for homeostasis and stem cell renewal. To distinguish oncogenic from physiological Wnt activity, we have performed transcriptome and proteome profiling in isogenic human colon organoids. Culture in the presence or absence of exogenous ligand allowed us to discriminate receptor-mediated signaling from the effects of CRISPR/Cas9-induced APC loss. We could catalog two nonoverlapping molecular signatures that were stable at distinct levels of stimulation. Newly identified markers for normal stem/progenitor cells and adenomas were validated by immunohistochemistry and flow cytometry. We found that oncogenic Wnt signals are associated with good prognosis in tumors of the consensus molecular subtype 2 (CMS2). In contrast, receptor-mediated signaling was linked to CMS4 tumors and poor prognosis. Together, our data represent a valuable resource for biomarkers that allow more precise stratification of Wnt responses in CRC.
Problematisation: The credibility and transparency of industrial and organisational psychological (IOP) research within South Africa was recently challenged by Efendic and Van Zyl (2019). The authors briefly showed inconsistencies in statistical results reported by authors of the South African Journal of Industrial Psychology (SAJIP), that various studies were insufficiently powered, that best-practice guidelines for the reporting of results were mostly only partially followed and that no transparency exists with regard to the research process. They demonstrated that authors of the SAJIP may knowingly or unknowingly be engaging in questionable research practices, which directly affects the credibility of both the discipline and the journal. Furthermore, they suggested practical guidelines for both authors and the SAJIP on how this could be managed.
Implications: Based on these suggestions, the authors invited prominent members of the IOP scientific community to provide scholarly commentary on their paper in order to aid in the development of ‘a clear strategy on how [the confidence crisis in IOP] could be managed, what the role of SAJIP is in this process and how SAJIP and its contributors could proactively engage to address these issues’. Seven members of the editorial board and two international scholars provided commentaries in an attempt to further the debate about the nature, causes, consequences and management of the credibility crisis within the South African context.
Purpose: The purpose of this final rebuttal article was to summarise and critically reflect on the commentaries of the nine articles to advance the debate on the confidence crisis within the South African IOP discipline.
Recommendations: All SAJIP’s stakeholders (authors, editors, reviewers, the publication house, universities and the journal) can play an active role in enhancing the credibility of the discipline. It is suggested that SAJIP should develop a clear and structured strategy to promote credible, transparent and ethical research practices within South Africa.
Background: Persistent antiphospholipid antibodies (aPL) constitute the serological hallmark of the antiphospholipid syndrome (APS). Recently, various new assay technologies for the detection of aPL better suited to multiplex reaction environments than ELISAs emerged. We evaluated the diagnostic performance of such a novel line immunoassay (LIA) for the simultaneous detection of 10 different aPL.
Methods: Fifty-three APS patients and 34 healthy controls were investigated for criteria (antibodies against cardiolipin [aCL], β2-glycoprotein I [aβ2-GPI]) and non-criteria aPL (antibodies against phosphatidic acid [aPA], phosphatidyl-choline [aPC], -ethanolamine [aPE], -glycerol [aPG], -inositol [aPI], -serine [aPS], annexin V [aAnnV], prothrombin [aPT]) IgG and IgM by LIA. Criteria aPL were additionally determined with the established Alegria (ALE), AcuStar (ACU), UniCap (UNI), and AESKULISA (AES) systems and non-criteria aPL with the AES system. Diagnostic performance was evaluated with a gold standard for criteria aPL derived from the results of the four established assays via latent class analysis and with the clinical diagnosis as gold standard for non-criteria aPL.
Results: Assay performance of the LIA for criteria aPL was comparable to that of ALE, ACU, UNI, and AES. For non-criteria aPL, sensitivities of the LIA for aPA-, aPI-, aPS-IgG and aPA-IgM were significantly higher and for aPC-, aPE-, aAnnV-IgG and aPC- and aPE-IgM significantly lower than AES. Specificities did not differ significantly.
Conclusions: The LIA constitutes a valuable diagnostic tool for aPL profiling. It offers increased sensitivity for the detection of aPL against anionic phospholipids. In contrast, ELISAs exhibit strengths for the sensitive detection of aPL against neutral phospholipids.
Uncontrolled constitutive activation of Wnt signaling is a hallmark of colorectal cancer (CRC), which is responsible for the initiation of the vast majority of CRC cases (Fearon and Vogelstein, 1990; Morin et al., 1997; Wood et al., 2007). Paneth cells support the small intestinal stem cells by providing them with the required niche factors and especially Wnt3. Although the normal colonic epithelium does not contain Paneth cells, Paneth cell metaplasia is frequently observed in human and mouse adenoma (Joo et al., 2009). The occurrence of Paneth cells suggests the presence of high levels of Wnt ligands with unknown function in the tumor microenvironment of Wnt-independent tumor cells. Tumor progression is recognized as result of evolving crosstalk between tumor cells and their surrounding non-transformed stromal cells (Hanahan and Weinberg, 2011; Wang et al., 2017). Although Wnt signaling has been intensively studied in colorectal cancer (CRC) cells (Zhan et al., 2017), it remains unclear whether Wnt activity in the tumor-associated stroma contributes to the tumor malignancy. The present thesis used the organoid 3D cell culture system, genetically modified mouse models as well as next generation sequencing technology to identify and characterise the role of Wnt signaling in the tumor microenvironment of CRC.
This introduction outlines new developments in the field of cultural and media memory studies in the wake of the transcultural turn. It pays specific attention to the twofold dynamics of memory’s travel and locatedness. While in recent memory studies discourse there has been a tendency to see travel as the inspiration for innovative research, locatedness has become associated with old-fashioned, bounded approaches. Rather than reproduce the positive charging of travel and negative charging of locatedness, this special issue aims to emphasise the complexity of memory dynamics resulting from the interaction of the two poles and to make visible that the production, (re)mediation, and reception of the past in the present is constituted by both travel and locatedness.
Summary
Wild relatives of crops thrive in habitats where environmental conditions can be restrictive for productivity and survival of cultivated species. The genetic basis of this variability, particularly for tolerance to high temperatures, is not well understood. We examined the capacity of wild and cultivated accessions to acclimate to rapid temperature elevations that cause heat stress (HS).
We investigated genotypic variation in thermotolerance of seedlings of wild and cultivated accessions. The contribution of polymorphisms associated with thermotolerance variation was examined regarding alterations in function of the identified gene.
We show that tomato germplasm underwent a progressive loss of acclimation to strong temperature elevations. Sensitivity is associated with intronic polymorphisms in the HS transcription factor HsfA2 which affect the splicing efficiency of its pre‐mRNA. Intron splicing in wild species results in increased synthesis of isoform HsfA2‐II, implicated in the early stress response, at the expense of HsfA2‐I which is involved in establishing short‐term acclimation and thermotolerance.
We propose that the selection for modern HsfA2 haplotypes reduced the ability of cultivated tomatoes to rapidly acclimate to temperature elevations, but enhanced their short‐term acclimation capacity. Hence, we provide evidence that alternative splicing has a central role in the definition of plant fitness plasticity to stressful conditions.
Cysteinyl leukotriene receptor 1 antagonists (CysLT1RA) are frequently used as add-on medication for the treatment of asthma. Recently, these compounds have shown protective effects in cardiovascular diseases. This prompted us to investigate their influence on soluble epoxide hydrolase (sEH) and peroxisome proliferator activated receptor (PPAR) activities, two targets known to play an important role in CVD and the metabolic syndrome. Montelukast, pranlukast and zafirlukast inhibited human sEH with IC50 values of 1.9, 14.1, and 0.8 μM, respectively. In contrast, only montelukast and zafirlukast activated PPARγ in the reporter gene assay with EC50 values of 1.17 μM (21.9% max. activation) and 2.49 μM (148% max. activation), respectively. PPARα and δ were not affected by any of the compounds. The activation of PPARγ was further investigated in 3T3-L1 adipocytes. Analysis of lipid accumulation, mRNA and protein expression of target genes as well as PPARγ phosphorylation revealed that montelukast was not able to induce adipocyte differentiation. In contrast, zafirlukast triggered moderate lipid accumulation compared to rosiglitazone and upregulated PPARγ target genes. In addition, we found that montelukast and zafirlukast display antagonistic activities concerning recruitment of the PPARγ cofactor CBP upon ligand binding suggesting that both compounds act as PPARγ modulators. In addition, zafirlukast impaired the TNFα triggered phosphorylation of PPARγ2 on serine 273. Thus, zafirlukast is a novel dual sEH/PPARγ modulator representing an excellent starting point for the further development of this compound class.
Persistent and, in particular, neuropathic pain is a major healthcare problem with still insufficient pharmacological treatment options. This triggered research activities aimed at finding analgesics with a novel mechanism of action. Results of these efforts will need to pass through the phases of drug development, in which experimental human pain models are established components e.g. implemented as chemical hyperalgesia induced by capsaicin. We aimed at ranking the various readouts of a human capsaicin–based pain model with respect to the most relevant information about the effects of a potential reference analgesic. In a placebo‐controlled, randomized cross‐over study, seven different pain‐related readouts were acquired in 16 healthy individuals before and after oral administration of 300 mg pregabalin. The sizes of the effect on pain induced by intradermal injection of capsaicin were quantified by calculating Cohen's d. While in four of the seven pain‐related parameters, pregabalin provided a small effect judged by values of Cohen's d exceeding 0.2, an item categorization technique implemented as computed ABC analysis identified the pain intensities in the area of secondary hyperalgesia and of allodynia as the most suitable parameters to quantify the analgesic effects of pregabalin. Results of this study provide further support for the ability of the intradermal capsaicin pain model to show analgesic effects of pregabalin. Results can serve as a basis for the designs of studies where the inclusion of this particular pain model and pregabalin is planned.
A review will be presented on the algebraic extension of the standard Teory of Relativity (GR) to the pseudocomplex formulation (pc-GR). Te pc-GR predicts the existence of a dark energy outside and inside the mass distribution, corresponding to a modifcation of the GR-metric. Te structure of the emission profle of an accretion disc changes also inside a star. Discussed are the consequences of the dark energy for cosmological models, permitting diferent outcomes on the evolution of the universe.