63 research outputs found

    Vergleichende Untersuchungen über die Entwicklung und Kompartimentierung der sakralen und kokzygealen Somiten im Hühnerembryo

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    Die vorliegende Arbeit befasst sich mit der Bildung und Reifung der sakralen und kokzygealen Somiten. Es handelt sich um eine systematische Übersicht, welche den Zeitpunkt des epithelio-mesenchymalen Übergangs in morphologischer und molekularer Sicht darstellt. Zum einen wurde die Morphologie des zunächst epithelialen und später kompartimentierten Somiten in semidünnen Schnitten untersucht. Zum anderen wurden molekulare Marker des Sklerotoms und des Myotoms dargestellt, die den Zeitpunkt der Kompartimentierung festlegen. Das Kompartiment Sklerotom wurde durch In situ-Hybridisierung mit dem Gen Pax1 dargestellt, das Myotom durch die Gene MyoD und Myf5, sowie durch das Protein MF20 ergänzt. Es wurden die Stadien HH 17 bis HH 27 (nach Hamburger und Hamilton) untersucht, wobei im Stadium HH 17, 18 und 19 sakrale Somiten gebildet werden und in den Stadien HH 19, 20, 21, 22 und 23 kokzygeale Somiten. Morphologie: Der zunächst noch epitheliale Somit entwickelt sich im Somitenstadium III bzw. IV in seine Kompartimente Sklerotom und Dermomyotom. Im Stadium HH 24 vollzieht sich der epithelio-mesenchymale Übergang bei Somit II. Ab Stadium HH 25 sind alle Somiten vollständig kompartimentiert. Als Besonderheit wurde eine nicht vollständige Epithelialisierung bei den Somiten I und II der Stadien HH 18 und 19 festgestellt. Sklerotom-Darstellung durch Pax1: Die Expression von Pax1 erscheint zunächst bei Somitenstadium III. Ab dem Stadium HH 22 erscheint sie schon bei Somit II; ab Stadium HH 25 enthalten alle Somiten Pax1-exprimierende Zellen. Die Expression zeichnet sich zunächst im dorsoventralen Sklerotomwinkel ab und weitet sich dann nach lateral aus. Myotom-Darstellung durch MyoD: Die ersten MyoD-exprimierenden Zellen treten bei den Somitenstadien III bzw. IV auf. Hier befindet sich die Expression zuerst im medialen Anteil und erstreckt sich später nach lateral, direkt ventral des Dermomyotoms. Ab dem Stadium HH 23 erscheint die MyoD-Expression des Myotoms bei Somit II. Ab Stadium HH 24 enthalten alle Somiten MyoD-exprimierende Zellen. Myotom-Darstellung durch Myf5: Myf5 wird in den Stadien HH 17 und HH 18 bei Somit III erstmals exprimiert. Die Stadien HH 19 und HH 22 bis HH 24 enthalten Myf5-exprimierende Zellen bei Somitenstadium II. Das Somitenstadium IV bei HH 20 und HH 21 zeigt eine Myf5-Expression. Ab HH 25 enthalten alle Somiten Myf5-exprimierende Zellen. Die Untersuchungen ergeben, dass der Prozess der Bildung und Reifung der kaudalen Somiten in gleicher Weise verläuft wie bei den kranialen Somiten. Jedoch vollzieht sich der epithelio-mesenchymale Übergang im kaudalen Embryoanteil ein Somitenstadium früher. Hier entwickeln sich die Somiten also schneller. Die Somitogenese verläuft beschleunigt ab.The presented study explores the formation and maturation of sacral and coccygeal somites. It provides a systematic overview of the time of the epithelio-mesenchymale transition from both a morphological and a molecular method point of view. On the one hand, the morphology of the firstly epithelial and then subdivided somite was analysed in semithinn-slides. On the other hand, molecular markers of compartments were used to indicate when compartmentalization starts. The compartments sclerotome and dermomytome were observed using in in situ-hybridisation with the genes Pax1, and MyoD and Myf5, respectivly; additionally, MF20-analysis was performed. The developmental stages of chick embryo (characterized by Hamburger and Hamilton) were explored from HH 17 to HH 27, whereas sacral somites are formed in HH 17, 18 and 19, coccygeal somites in HH 19, 20, 21, 22 and 23. Morphology: The epithelial somite develops into compartments sclerotome and dermomyotome at the stage of somite III and IV. At HH 24 the epithelio-mesenchymal transition occurs at stage of somite II; from HH 25 onwards all of the somites are transformed completely. It is worth noting, that at HH 18 and 19 the formation of the epithelial ball of somites I and II is incomplete. Labelling of sclerotome by Pax1: Pax1 is expressed at stage of somite III for the first time. From HH 22, the expression appears at somite II; from HH 25 all somites include cells that express Pax1. Signaling begins at the dorsoventral angle of the sclerotome and extends laterally later on. Labelling of myotome by MyoD: The first cells who express MyoD appear at stage of somite III and IV. At first, signaling can be observed at the medial part and then extends laterally, ventral of the dermomyotome. At HH 23, signaling of the myotome occurs at stage of somite II; from HH 24 all somites show expression of MyoD. Labelling of myotome by Myf5: At HH 17 and HH 18, Myf5 is expressed for the first time at stage of somite III by in situ-hybridisation. At HH 19 and from HH 22 to 24 Myf5-expression appears at stage of somite II. Stage of somite IV expresses Myf5 between HH 20 and 21. From HH 25 all somites show expression of Myf5. In conclusion, the study shows that the developments of caudal and of cranial somites happen in the same way. However, the epithelio-to-mesenchymal transition in the caudal region occurs one stage of somite earlier; thus demonstrating the caudal development of somites is faster than the cranial development of somites

    Performance in Sound-Symbol Learning Predicts Reading Performance 3 Years Later

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    To master the task of reading, children need to acquire a coding system representing speech as a sequence of visual symbols. Recent research suggested that performance in the processing of artificial script that relies on the association of sound and symbol may be associated with reading skill. The current longitudinal study examined the predictive value of a preschool sound-symbol paradigm (SSP) of reading performance 3 years later. The Morse-like SSP, IQ, and letter knowledge (LK) was assessed in young preschool children. Reading outcome measures were examined 3 years later. Word reading, pseudoword reading, and reading comprehension were predicted with age, IQ, LK, and SSP. The results showed that SSP substantially predicted reading fluency and reading comprehension 3 years later. For reading fluency measures, the influence of further predictor variables was not significant and SSP served as a sole predictor. Reading comprehension was best explained by SSP and age. The amount of variance SSP explained in reading 3 years later was remarkably high, with an explained variance between 63 and 82%, depending on the outcome reading variable. SSP turned out to be a substantial predictor of later reading performance in a language with statistically reliable spelling-to-sound relations. As LK is highly dependent on educational support, we assume that children in our socioeconomically diverse sample did not have much opportunity to acquire LK in their home environment. In contrast, the SSP challenges students to acquire new spelling-to-sound relations, simulating a core aspect of natural reading acquisition. Future work will test this paradigm in less transparent languages like English and explore its potential as a future standard assessment in the study of early reading development

    TOX defines the degree of CD8+ T cell dysfunction in distinct phases of chronic HBV infection

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    Objective Chronic hepatitis B virus (HBV) infection is characterised by HBV-specific CD8+ T cell dysfunction that has been linked to Tcell exhaustion, a distinct differentiation programme associated with persisting antigen recognition. Recently, Thymocyte Selection-Associated High Mobility Group Box (TOX) was identified as master regulator of CD8+ T cell exhaustion. Here, we addressed the role of TOX in HBV-specific CD8+ T cell dysfunction associated with different clinical phases of infection. Design We investigated TOX expression in HBV-specific CD8+ T cells from 53 HLA-A*01:01, HLA-A*11:01 and HLA-A*02:01 positive patients from different HBV infection phases and compared it to hepatitis C virus (HCV)-specific, cytomegalovirus (CMV)-specific, Epstein-Barr virus (EBV)-specific and influenza virus (FLU)-specific CD8+ T cells. Phenotypic and functional analyses of virus-specific CD8+ T cells were performed after peptide-loaded tetramer-enrichment and peptide-specific expansion. Results Our results show that TOX expression in HBV-specific CD8+ T cells is linked to chronic antigen stimulation, correlates with viral load and is associated with phenotypic and functional characteristics of T-cell exhaustion. In contrast, similar TOX expression in EBV-specific and CMV-specific CD8+ T cells is not linked to T-cell dysfunction suggesting different underlying programmes. TOX expression in HBV-specific CD8+ T cells is also affected by targeted antigens, for example, core versus polymerase. In HBV-specific CD8+ T cells, TOX expression is maintained after spontaneous or therapy-mediated viral control in chronic but not self-limiting acute HBV infection indicating a permanent molecular imprint after chronic but not temporary stimulation. Conclusion Our data highlight TOX as biomarker specific for dysfunctional virus-specific CD8+ T cells in the context of an actively persisting infection

    Phenotypic and functional differences of HBV core-specific versus HBV polymerase-specific CD8+ T cells in chronically HBV-infected patients with low viral load

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    Objective A hallmark of chronic HBV (cHBV) infection is the presence of impaired HBV-specific CD8+ T cell responses. Functional T cell exhaustion induced by persistent antigen stimulation is considered a major mechanism underlying this impairment. However, due to their low frequencies in chronic infection, it is currently unknown whether HBV-specific CD8+ T cells targeting different epitopes are similarly impaired and share molecular profiles indicative of T cell exhaustion. Design By applying peptide-loaded MHC I tetramer-based enrichment, we could detect HBV-specific CD8+ T cells targeting epitopes in the HBV core and the polymerase proteins in the majority of 85 tested cHBV patients with low viral loads. Lower detection rates were obtained for envelope-specific CD8+ T cells. Subsequently, we performed phenotypic and functional in-depth analyses. Results HBV-specific CD8+ T cells are not terminally exhausted but rather exhibit a memory-like phenotype in patients with low viral load possibly reflecting weak ongoing cognate antigen recognition. Moreover, HBV-specific CD8+ T cells targeting core versus polymerase epitopes significantly differed in frequency, phenotype and function. In particular, in comparison with core-specific CD8+ T cells, a higher frequency of polymerase-specific CD8+ T cells expressed CD38, KLRG1 and Eomes accompanied by low T-bet expression and downregulated CD127 indicative of a more severe T cell exhaustion. In addition, polymerase-specific CD8+ T cells exhibited a reduced expansion capacity that was linked to a dysbalanced TCF1/BCL2 expression. Conclusions Overall, the molecular mechanisms underlying impaired T cell responses differ with respect to the targeted HBV antigens. These results have potential implications for immunotherapeutic approaches in HBV cure

    CD169/SIGLEC1 is expressed on circulating monocytes in COVID-19 and expression levels are associated with disease severity

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    Coronavirus disease 2019 (COVID-19) is caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Type I interferons are important in the defense of viral infections. Recently, neutralizing IgG auto-antibodies against type I interferons were found in patients with severe COVID-19 infection. Here, we analyzed expression of CD169/SIGLEC1, a well described downstream molecule in interferon signaling, and found increased monocytic CD169/SIGLEC1 expression levels in patients with mild, acute COVID-19, compared to patients with severe disease. We recommend further clinical studies to evaluate the value of CD169/SIGLEC1 expression in patients with COVID-19 with or without auto-antibodies against type I interferons

    Addiction Research Consortium: Losing and regaining control over drug intake (ReCoDe)—From trajectories to mechanisms and interventions

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    One of the major risk factors for global death and disability is alcohol, tobacco, and illicit drug use. While there is increasing knowledge with respect to individual factors promoting the initiation and maintenance of substance use disorders (SUDs), disease trajectories involved in losing and regaining control over drug intake (ReCoDe) are still not well described. Our newly formed German Collaborative Research Centre (CRC) on ReCoDe has an interdisciplinary approach funded by the German Research Foundation (DFG) with a 12-year perspective. The main goals of our research consortium are (i) to identify triggers and modifying factors that longitudinally modulate the trajectories of losing and regaining control over drug consumption in real life, (ii) to study underlying behavioral, cognitive, and neurobiological mechanisms, and (iii) to implicate mechanism-based interventions. These goals will be achieved by: (i) using mobile health (m-health) tools to longitudinally monitor the effects of triggers (drug cues, stressors, and priming doses) and modify factors (eg, age, gender, physical activity, and cognitive control) on drug consumption patterns in real-life conditions and in animal models of addiction; (ii) the identification and computational modeling of key mechanisms mediating the effects of such triggers and modifying factors on goal-directed, habitual, and compulsive aspects of behavior from human studies and animal models; and (iii) developing and testing interventions that specifically target the underlying mechanisms for regaining control over drug intake

    Patterns of Alcohol Consumption Among Individuals With Alcohol Use Disorder During the COVID-19 Pandemic and Lockdowns in Germany

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    Importance Alcohol consumption (AC) leads to death and disability worldwide. Ongoing discussions on potential negative effects of the COVID-19 pandemic on AC need to be informed by real-world evidence. Objective To examine whether lockdown measures are associated with AC and consumption-related temporal and psychological within-person mechanisms. Design, Setting, and Participants This quantitative, intensive, longitudinal cohort study recruited 1743 participants from 3 sites from February 20, 2020, to February 28, 2021. Data were provided before and within the second lockdown of the COVID-19 pandemic in Germany: before lockdown (October 2 to November 1, 2020); light lockdown (November 2 to December 15, 2020); and hard lockdown (December 16, 2020, to February 28, 2021). Main Outcomes and Measures Daily ratings of AC (main outcome) captured during 3 lockdown phases (main variable) and temporal (weekends and holidays) and psychological (social isolation and drinking intention) correlates. Results Of the 1743 screened participants, 189 (119 [63.0%] male; median [IQR] age, 37 [27.5-52.0] years) with at least 2 alcohol use disorder (AUD) criteria according to the Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition) yet without the need for medically supervised alcohol withdrawal were included. These individuals provided 14 694 smartphone ratings from October 2020 through February 2021. Multilevel modeling revealed significantly higher AC (grams of alcohol per day) on weekend days vs weekdays (β = 11.39; 95% CI, 10.00-12.77; P < .001). Alcohol consumption was above the overall average on Christmas (β = 26.82; 95% CI, 21.87-31.77; P < .001) and New Year’s Eve (β = 66.88; 95% CI, 59.22-74.54; P < .001). During the hard lockdown, perceived social isolation was significantly higher (β = 0.12; 95% CI, 0.06-0.15; P < .001), but AC was significantly lower (β = −5.45; 95% CI, −8.00 to −2.90; P = .001). Independent of lockdown, intention to drink less alcohol was associated with lower AC (β = −11.10; 95% CI, −13.63 to −8.58; P < .001). Notably, differences in AC between weekend and weekdays decreased both during the hard lockdown (β = −6.14; 95% CI, −9.96 to −2.31; P = .002) and in participants with severe AUD (β = −6.26; 95% CI, −10.18 to −2.34; P = .002). Conclusions and Relevance This 5-month cohort study found no immediate negative associations of lockdown measures with overall AC. Rather, weekend-weekday and holiday AC patterns exceeded lockdown effects. Differences in AC between weekend days and weekdays evinced that weekend drinking cycles decreased as a function of AUD severity and lockdown measures, indicating a potential mechanism of losing and regaining control. This finding suggests that temporal patterns and drinking intention constitute promising targets for prevention and intervention, even in high-risk individuals

    Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

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    A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

    Measurements of top-quark pair differential cross-sections in the eμe\mu channel in pppp collisions at s=13\sqrt{s} = 13 TeV using the ATLAS detector

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