CRISPRi screening reveals regulators of tau pathology shared between exosomal and vesicle-free tau

The aggregation of the microtubule-associated protein tau is a defining feature of Alzheimer's disease and other tauopathies. Tau pathology is believed to be driven by free tau aggregates and tau carried within exosome-like extracellular vesicles, both of which propagate trans-synaptically and induce tau pathology in recipient neurons by a corrupting process of seeding. Here, we performed a genome-wide CRISPRi screen in tau biosensor cells and identified cellular regulators shared by both mechanisms of tau seeding. We identified ANKLE2, BANF1, NUSAP1, EIF1AD, and VPS18 as the top validated regulators that restrict tau aggre-gation initiated by both exosomal and vesicle-free tau seeds. None of our validated hits affected the uptake of either form of tau seeds, supporting the notion that they operate through a cell -autonomous mechanism downstream of the seed uptake. Lastly, validation studies with human brain tissue also revealed that several of the identified protein hits are down-regulated in the brains of Alzheimer's patients, suggesting that their decreased activity may be required for the emergence or progression of tau pathology in the human brain.Peer reviewe

Association Between Intravenous Thrombolysis and Clinical Outcomes Among Patients With Ischemic Stroke and Unsuccessful Mechanical Reperfusion.

IMPORTANCE Clinical evidence of the potential treatment benefit of intravenous thrombolysis preceding unsuccessful mechanical thrombectomy (MT) is scarce. OBJECTIVE To determine whether intravenous thrombolysis (IVT) prior to unsuccessful MT improves functional outcomes in patients with acute ischemic stroke. DESIGN, SETTING, AND PARTICIPANTS Patients were enrolled in this retrospective cohort study from the prospective, observational, multicenter German Stroke Registry-Endovascular Treatment between May 1, 2015, and December 31, 2021. This study compared IVT plus MT vs MT alone in patients with acute ischemic stroke due to anterior circulation large-vessel occlusion in whom mechanical reperfusion was unsuccessful. Unsuccessful mechanical reperfusion was defined as failed (final modified Thrombolysis in Cerebral Infarction grade of 0 or 1) or partial (grade 2a). Patients meeting the inclusion criteria were matched by treatment group using 1:1 propensity score matching. INTERVENTIONS Mechanical thrombectomy with or without IVT. MAIN OUTCOMES AND MEASURES Primary outcome was functional independence at 90 days, defined as a modified Rankin Scale score of 0 to 2. Safety outcomes were the occurrence of symptomatic intracranial hemorrhage and death. RESULTS After matching, 746 patients were compared by treatment arms (median age, 78 [IQR, 68-84] years; 438 women [58.7%]). The proportion of patients who were functionally independent at 90 days was 68 of 373 (18.2%) in the IVT plus MT and 42 of 373 (11.3%) in the MT alone group (adjusted odds ratio [AOR], 2.63 [95% CI, 1.41-5.11]; P = .003). There was a shift toward better functional outcomes on the modified Rankin Scale favoring IVT plus MT (adjusted common OR, 1.98 [95% CI, 1.35-2.92]; P < .001). The treatment benefit of IVT was greater in patients with partial reperfusion compared with failed reperfusion. There was no difference in symptomatic intracranial hemorrhages between treatment groups (AOR, 0.71 [95% CI, 0.29-1.81]; P = .45), while the death rate was lower after IVT plus MT (AOR, 0.54 [95% CI, 0.34-0.86]; P = .01). CONCLUSIONS AND RELEVANCE These findings suggest that prior IVT was safe and improved functional outcomes at 90 days. Partial reperfusion was associated with a greater treatment benefit of IVT, indicating a positive interaction between IVT and MT. These results support current guidelines that all eligible patients with stroke should receive IVT before MT and add a new perspective to the debate on noninferiority of combined stroke treatment

Redirecting T Cells to Ewing's Sarcoma Family of Tumors by a Chimeric NKG2D Receptor Expressed by Lentiviral Transduction or mRNA Transfection

We explored the possibility to target Ewing's sarcoma family of tumors (ESFT) by redirecting T cells. To this aim, we considered NKG2D-ligands (NKG2D-Ls) as possible target antigens. Detailed analysis of the expression of MICA, MICB, ULBP-1, -2, and -3 in fourteen ESFT cell lines revealed consistent expression of at least one NKG2D-L. Thus, for redirecting T cells, we fused a CD3ζ/CD28-derived signaling domain to the ectodomain of NKG2D, however, opposite transmembrane orientation of this signaling domain and NKG2D required inverse orientation fusion of either of them. We hypothesized that the particularly located C-terminus of the NKG2D ectodomain should allow reengineering of the membrane anchoring from a native N-terminal to an artificial C-terminal linkage. Indeed, the resulting chimeric NKG2D receptor (chNKG2D) was functional and efficiently mediated ESFT cell death triggered by activated T cells. Notably, ESFT cells with even low NKG2D-L expression were killed by CD8pos and also CD4pos cells. Both, mRNA transfection and lentiviral transduction resulted in high level surface expression of chNKG2D. However, upon target-cell recognition receptor surface levels were maintained by tranfected RNA only during the first couple of hours after transfection. Later, target-cell contact resulted in strong and irreversible receptor down-modulation, whereas lentivirally mediated expression of chNKG2D remained constant under these conditions. Together, our study defines NKG2D-Ls as targets for a CAR-mediated T cell based immunotherapy of ESFT. A comparison of two different methods of gene transfer reveals strong differences in the susceptibility to ligand-induced receptor down-modulation with possible implications for the applicability of RNA transfection

The THESEUS space mission concept: science case, design and expected performances

THESEUS is a space mission concept aimed at exploiting Gamma-Ray Bursts for investigating the early Universe and at providing a substantial advancement of multi-messenger and time-domain astrophysics. These goals will be achieved through a unique combination of instruments allowing GRB and X-ray transient detection over a broad field of view (more than 1sr) with 0.5¿1 arcmin localization, an energy band extending from several MeV down to 0.3¿keV and high sensitivity to transient sources in the soft X-ray domain, as well as on-board prompt (few minutes) follow-up with a 0.7¿m class IR telescope with both imaging and spectroscopic capabilities. THESEUS will be perfectly suited for addressing the main open issues in cosmology such as, e.g., star formation rate and metallicity evolution of the inter-stellar and intra-galactic medium up to redshift 10, signatures of Pop III stars, sources and physics of re-ionization, and the faint end of the galaxy luminosity function. In addition, it will provide unprecedented capability to monitor the X-ray variable sky, thus detecting, localizing, and identifying the electromagnetic counterparts to sources of gravitational radiation, which may be routinely detected in the late ¿20s/early ¿30s by next generation facilities like aLIGO/ aVirgo, eLISA, KAGRA, and Einstein Telescope. THESEUS will also provide powerful synergies with the next generation of multi-wavelength observatories (e.g., LSST, ELT, SKA, CTA, ATHENA).© 2018 COSPARS.E. acknowledges the financial support from contracts ASI-INAF 1/009/10/0, NARO15 ASI-INAF 1/037/12/0 and ASI 2015-046-R.0. R.H. acknowledges GACR grant 13-33324S. S.V. research leading to these results has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no 606176. D.S. was supported by the Czech grant 1601116S GA CR. Maria Giovanna Dainotti acknowledges funding from the European Union through the Marie Curie Action FP7-PEOPLE-2013-IOF, under grant agreement No. 626267 (>Cosmological Candles>)

A multi-country test of brief reappraisal interventions on emotions during the COVID-19 pandemic.

The COVID-19 pandemic has increased negative emotions and decreased positive emotions globally. Left unchecked, these emotional changes might have a wide array of adverse impacts. To reduce negative emotions and increase positive emotions, we tested the effectiveness of reappraisal, an emotion-regulation strategy that modifies how one thinks about a situation. Participants from 87 countries and regions (n = 21,644) were randomly assigned to one of two brief reappraisal interventions (reconstrual or repurposing) or one of two control conditions (active or passive). Results revealed that both reappraisal interventions (vesus both control conditions) consistently reduced negative emotions and increased positive emotions across different measures. Reconstrual and repurposing interventions had similar effects. Importantly, planned exploratory analyses indicated that reappraisal interventions did not reduce intentions to practice preventive health behaviours. The findings demonstrate the viability of creating scalable, low-cost interventions for use around the world

Ökonomische Überlegungen zur theoretischen Wiedervereinigung Koreas : Auswirkungen auf umliegende Staaten, Währung und die Gesamtwirtschaft

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An examination of the influence of social changes (change of values) on the motivation of suicide

Titel und Inhaltsverzeichnis Einleitung Kapitel 1: Phänomenologie des Suizides Kapitel 2: Werte Kapitel 3: Suizidtheorien Kapitel 4: Das sozialpsychologische Dissonanz-Einengungskonstrukt Kapitel 5: Quantitative Untersuchungen Kapitel 6: Qualitative Untersuchung Kapitel 7: Hypothesenprüfung und Zusammenfassung der wichtigsten Ergebnisse Kapitel 8: Suizidforen: Neuland für die professionellen Helfer Kapitel 9: LiteraturDie spezifische Konzentration der Dissertation auf die gesellschaftlichen Entwicklungen in beiden deutschen Staaten ab 1961 bis zur Wende und darüber hinaus und auf den Zusammenhang dieser Entwicklungen mit der Suizidalität basiert auf der Annahme, dass sich der Einfluss gesellschaftlicher Veränderungsprozesse (Wertewandel) auf ein individuelles Geschehen wie den Suizid am besten an einem solchen gesellschaftlichen Großereignis studieren lässt. Im Mittelpunkt steht die regio-und geschlechtsspezifische Betrachtung des Suizidgeschehens bei der jungen Generation im Alter von 10 bis 25 Jahren. Im theoretischen Teil wird eine phänomenologische Skizze der verschiedenen Betrachtungsweisen und Erklärungen von Suizid, Suizidalität, parasuizidalen Handlungen vorgenommen, der eine Auseinandersetzung mit der Werteproblematik folgt. Dabei wird aus der Perspektive des lernenden Individuums die Begegnung mit Werten und Normen, Werteerziehung und Wertevermittlung dargestellt. Daran anschließend werden Formen von Wertewandel anhand historischer Beispiele beschrieben. Häufig in der Fachliteratur erwähnte Suiziderklärungsansätze sowie prädisponierende und protektive Faktoren werden im Anschluss vorgestellt. Angesichts der Tatsache, dass die beschriebenen Erklärungsansätze einzeln als Rahmentheorie unzulänglich sind, erfolgt der Entwurf eines Handlungsmodells in Form eines Personen-Umwelt-Interaktionsansatzes. Zur Durchführung der Untersuchung werden zwei Methoden kombiniert. Als erstes erfolgt eine quantitative Längschnittuntersuchung, basierend auf Daten aus amtlichen Statistiken, die das Suizidgeschehen in Ost und West zwischen 1961 und 2005 in seinem quantitativen Verlauf analysiert. Im Anschluss folgt eine Analyse der Fachliteratur der DDR und früheren Bundesrepublik. Die qualitative Untersuchung basiert auf Ergebnissen von 27 durchgeführten leitfadengestützten Experteninterviews. Die verschiedenen Teile der empirischen Untersuchung werden anschließend zusammengeführt. Die quantitative Untersuchung der Suizidraten zeigt eine anhaltende Konstanz höherer Suizidmortalität in der DDR im Vergleich zur früheren Bundesrepublik. Im Verlauf der 80er Jahre ist in beiden Gebieten ein deutlicher Rückgang der Suizidraten festzustellen, der sich über die Wiedervereinigung hinaus fortsetzt und im Zusammenhang mit allmählichen gesellschaftlichen Wandlungsprozessen zu sehen ist, die sich in Form von veränderten Verhaltensbereitschaften und geändertem Wertinteresse konkretisieren. Vor dem Hintergrund sich angleichender gesellschaftlicher Rahmenbedingungen nach der Wiedervereinigung nähern sich die Suizidraten in Ost und West an. Es kann festgestellt werden, dass mit dem Großereignis der Wende nur vorübergehend ein Anstieg der Suizidhäufigkeit bei der jungen Bevölkerung in den neuen Bundesländern verbunden ist. Die Zusammenführung beider Gebiete Deutschlands in Form eines Transformationsprozesses an die westlichen Werte-und Normenfeststellungen wurde von der jungen Generation in der ehemaligen DDR hingegen früherer Vorhersagungen offensichtlich besser verkraftet. Die qualitativen Untersuchungsergebnisse bestätigen Zusammenhänge zwischen Wertewandel und Suizidgeschehen und weisen auch auf die Bedeutung von Werten und Wertevermittlung hin. Darüber hinaus berichten die Interviewten Einstellungsveränderungen zur Suizidthematik.The specific emphasis of the dissertation, the development of society in the two German nations since 1961 until the unification and beyond and apparent relationship to suicidality, is based on the assumption that the influence of social changes in society (change of values) on individual behavior like suicide, can best be researched from the view point of such key event in society. The focus has been on suicide mortality by region and gender of the young generation, age 10 to 25. The theoretical part sketches out the different points of view and explanations on suicide, suicidality, para- suicidal actions and is followed by a discussion on value issues. It presents a perspective of the learning individual facing values, norms as well as education on values and the passing on of sets of values. Thereafter, cases of social change are presented in a historical context. It follows a presentation often used suicide theories in expert literature of suicide like pre- disposition and protective factors. Due to the fact that the described explanation attempts are not valid individually as a theoretical framework, a design of a theory has been created based on an individual-environment interaction model. Two methods have been applied in the research. A quantitative analysis was done based on public data on suicide in East and West between 1961 and 2005, followed by an analysis of expert literature in the former GDR and FRG. The qualititative part was derived from the results of 27 expert interviews conducted by way of specific questionnaires. Then, different empirical parts of the research have been combined. The quantitative research of suicide rates points to a consistently higher degree of suicide mortality in the GDR compared to the FRG data. During the 1980s, the suicide mortality rates show a sharp decrease in both regions. This trend has continued after unification due to gradual social changes resulting from alterations of behaviors and interest in values. As a consequence of the alignment of the framework after unification, the suicide rates in East and West have come closer. Findings show that there was only a temporary increase of suicide mortality among the young population of the new German states after unification. Contrary to former predictions, the young generation of the former GDR has apparently been coping much better with the unification of the two german nations, done through a transformation process towards western value standards. The qualititative research results confirm the relationship between social change (change of values) and suicide rates and furthermore point to the importance of values the passing on of them. More importantly, the interviewees report changes in attitudes towards the subject of suicide

Rodent models for Alzheimer disease

Animal models are indispensable tools for Alzheimer disease (AD) research. Over the course of more than two decades, an increasing number of complementary rodent models has been generated. These models have facilitated testing hypotheses about the aetiology and progression of AD, dissecting the associated pathomechanisms and validating therapeutic interventions, thereby providing guidance for the design of human clinical trials. However, the lack of success in translating rodent data into therapeutic outcomes may challenge the validity of the current models. This Review critically evaluates the genetic and non-genetic strategies used in AD modelling, discussing their strengths and limitations, as well as new opportunities for the development of better models for the disease

Three methods for examining the de novo proteome of microglia using BONCAT bioorthogonal labeling and FUNCAT click chemistry

Summary: Bioorthogonal labeling and click chemistry techniques allow the detailed examination of cellular physiology through tagging and visualizing newly synthesized proteins. Here, we describe three methods applying bioorthogonal non-canonical amino acid tagging and fluorescent non-canonical amino acid tagging to quantify protein synthesis in microglia. We describe steps for cell seeding and labeling. We then detail microscopy, flow cytometry, and Western blotting techniques. These methods can be easily adapted for other cell types to explore cellular physiology in health and disease.For complete details on the use and execution of this protocol, please refer to Evans et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics

Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation

The formation of spatial long-term memory (LTM) requires the de novo synthesis of distinct sets of proteins; however, a non-biased examination of the de novo proteome in this process is lacking. Here, we generated a novel mouse strain, which enables cell-type-specific labelling of newly synthesised proteins with non-canonical amino acids (NCAAs) by genetically restricting the expression of the mutant tRNA synthetase, NLL-MetRS, to hippocampal neurons. By combining this labelling technique with an accelerated version of the active place avoidance task and bio-orthogonal non-canonical amino acid tagging (BONCAT) followed by SWATH quantitative mass spectrometry, we identified 156 proteins that were altered in synthesis in hippocampal neurons during spatial memory formation. In addition to observing increased synthesis of known proteins important in memory-related processes, such as glutamate receptor recycling, we also identified altered synthesis of proteins associated with mRNA splicing as a potential mechanism involved in spatial LTM formation