Expression, purification and functional validation of a cancer-associated isoform of the HBx protein from human hepatitis B virus

The human hepatitis B virus (HBV) causes hepatitis B, a liver infection that can be acute or chronic. HBV encodes four proteins, among which the X protein (HBx) plays a critical role in viral replication. During chronic HBV infection, in which the viral DNA is integrated into the host genome, the HBx1-120 isoform, comprising the N-terminal 120 residues, is highly expressed. Here, we describe a protocol for the recombinant overexpression and purification of untagged HBx1-120 from bacterial cells. The procedure is compatible with stable isotope labelling in minimal media. Following cell lysis, HBx1-120 was recovered from inclusion bodies (IBs), solubilized in urea, and purified by ion-exchange (IEX) and size-exclusion chromatography (SEC). The purified protein was extensively characterized, including by mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. Functionality was confirmed by a pulldown assay with a known interacting partner, Spindlin1. This protocol provides a robust framework to obtain untagged HBx1-120 for structural and functional in vitro studies

Predicting pacemaker dependency after transcatheter aortic valve replacement: Analysis of ECG, clinical, and CT-imaging parameters

Objectives Conduction disturbances necessitating permanent pacemaker (PPM) implantation following transcatheter aortic valve replacement (TAVR) have been observed. However, limited data exist on ECG, clinical, and CT-imaging factors predicting PPM dependency after TAVR. This study aimed to identify predictors of pacemaker dependency in selected patients who required PPM implantation after TAVR with SAPIEN 3 prostheses. Materials and Methods This study included consecutive patients who underwent transfemoral TAVR with SAPIEN 3 prostheses at our institution between May 2012 and December 2019. Exclusion criteria were incomplete or non-diagnostic data, valve-in-valve procedures, TAVR in mitral position, previous surgical repair, and pre-implanted PPM. The primary endpoint was PPM dependency, defined as ventricular pacing percentage ≥ 1 % at the first outpatient follow-up after PPM implantation post-TAVR. Regression analysis was performed to compare a limited prediction model for PPM dependency using only selected variables to a full model with all available variables. Results Out of 2105 patients who received TAVR, 350 (16.6 %) required pacemaker implantation post-TAVR. After exclusions, 301 patients remained, with 168 (55.8 %) PPM-dependent and 133 (44.2 %) non-dependent patients. Multivariate analysis identified prosthesis oversizing (OR: 1.09, p < 0.001), calcification below the left coronary cusp (LCC) (OR: 1.02, p < 0.001), and right bundle branch block (RBBB) prior to TAVR (OR: 2.20, p = 0.025) as significant predictors. A limited regression model predicted PPM dependency with an AUC of 0.752, significantly outperforming the full model (AUC: 0.660, p = 0.037). Conclusion RBBB prior to TAVR was the strongest predictor of PPM dependency post-TAVR, followed by prosthesis oversizing and calcification below the LCC. A limited prediction model with these variables demonstrated moderate predictive accuracy

High-stress crystal plasticity of titanite and other minerals recording coseismic off-fault damage in strike-slip and thrust tectonic settings

This study compares coseismic off-fault damage in a strike-slip (Sesia zone) and a thrust (Silvretta) tectonic setting to evaluate the different stress-strain histories. High-stress crystal plasticity at greenschist-facies conditions is recorded by mylonites from the Sesia zone and pseudotachylyte-bearing gneisses from the Silvretta basal thrust, European Alps. Twinned titanite occurring in both fault rocks highlights the similarities and differences in the recorded deformation. Fine-lamellar ( twins in titanite from the Sesia mylonites with twin planes close to {221} show densities of 0.5 µm−1. Consistent with twinned jadeite, the differential stresses indicated are on the order of 0.5 GPa. In the Silvretta fault rocks, the twin density is higher with 2.5 µm−1 and additionally, twin planes close to {112} occur, indicating higher stress/strain-rate conditions, consistent with twinned amphibole and ilmenite as well as the presence of pseudotachylytes. The Silvretta fault rocks do not record subsequent creep, indicating rapidly decreasing stresses. In contrast, in the Sesia mylonites, subsequent creep of the surrounding quartz matrix at decreasing stresses resulted in sets of subparallel intragranular fractures in titanite, garnet, jadeite and zircon oriented at angles between 60° and 80° to the mylonitic foliation. The similarities of high-stress crystal plasticity in both settings with twinning at high differential stresses, as well as the differences with pseudotachylyte formation in the Silvretta fault rocks and creep at more slowly decreasing stresses in the Sesia zone mylonites, demonstrate the importance of deformation at transient high stresses for the subsequent stress-strain history

In vivo synergistic enhancement of MIF‐mediated inflammation in acute lung injury by the plant ortholog ArabidopsisMDL1

Recent research has uncovered Arabidopsis thaliana proteins that are similar to the human inflammatory cytokine MIF. Plant MIF/D-dopachrome tautomerase (D-DT)-like proteins (MDLs) can interact with human MIF, yet the significance of these findings in living organisms has not been investigated. Given MIF's key role in acute respiratory distress syndrome promoting pulmonary inflammation, pathology, and leukocyte infiltration, here we set out to investigate the interplay between MIF and MDL1, one of three A. thaliana MIF orthologs, in an in vivo mouse model of MIF-induced acute lung injury (ALI). Human MIF and MDL1 were administered to C57BL/6 mice via inhalation, individually or in combination. Inhalation of MIF promoted various parameters of lung injury as evaluated by flow cytometry, immunofluorescence microscopy, RT-qPCR, and ELISA, while MDL1 inhalation alone had no effect. Intriguingly, combined treatment with MIF and MDL1 synergistically enhanced pulmonary infiltration of neutrophils and monocytic cells, accompanied by an upregulation of pro-inflammatory cytokine genes. Thus, the plant-derived MIF ortholog MDL1 potentiates MIF-induced inflammation in ALI. These data support the growing evidence of interactions between plant-derived compounds and human inflammatory mediators and illustrate how they may impact human health

Learning regular cross-trial shifts of the target location in serial search involves awareness – An eye-tracking study

People can learn and use both static and dynamic (cross-trial) regularities in the positioning of target items during parallel, ‘pop-out’ visual search. Static target-location learning also works in serial search, however, acquiring dynamic regularities is hindered by the demands of item-by-item scanning. Also, questions have been raised regarding whether explicit awareness is necessary for using dynamic regularities to optimize performance. The present study re-examined if dynamic regularities can be learned in serial search when regular shifts of the target location occur frequently, and if such learning correlates with awareness of the dynamic rule. We adopted the same regularity used by Yu et al. (2023) to demonstrate dynamic learning in parallel search: a cross-trial shift of the target location in a (counter-)clockwise direction within a circular array in 80 % of the trials, compared to irregular shifts in the opposite direction (10 %) or some other random direction (10 %). The results showed that about 70 % of participants learned the dynamic regularity, with performance gains correlating with awareness: the more accurately they estimated the likelihood of the target shifting in the frequent direction, the greater their gains. Importantly, part of the gains accrued already early during the search: a large proportion of the very first and short-latency eye movements were directed to the predicted location, regardless of the target appeared there. We discuss whether this rule-driven behavior is causally mediated by conscious control

Impact of Pulmonary Hypertension on Outcomes After Transcatheter Tricuspid Valve Edge-to-Edge Repair

Background Data regarding the association of pulmonary hypertension (PH) and outcomes in patients undergoing transcatheter tricuspid valve edge-to-edge repair (T-TEER) are scarce. Objectives The aims of this study were: 1) to investigate the impact of PH on outcomes after T-TEER; and 2) to shed further light on the role of precapillary- and postcapillary PH in patients undergoing T-TEER for relevant tricuspid regurgitation (TR). Methods The study included patients from EuroTR (European Registry of Transcatheter Repair for Tricuspid Regurgitation; NCT06307262) who underwent T-TEER for relevant TR from 2016 until 2023 with available invasive evaluation of systolic pulmonary artery pressure (sPAP) using right heart catheterization. Study endpoints were procedural TR reduction, improvement in NYHA function class, and a combined endpoint of death or heart failure hospitalization (HFH) at 2 years. Results Among a total of 1,230 patients (mean age 78.6 ± 7.0 years, 51.4% women), increasing sPAP was independently associated with increasing rates of 2-year death or HFH (HR: 1.027; 95% CI: 1.003-1.052; P = 0.030; median survival follow-up 343 days [Q1-Q3: 114-645 days]). No significant survival differences were observed for patients with pre- vs postcapillary PH. Sensitivity analysis revealed an sPAP value of 46 mm Hg as the optimized threshold for the prediction of death or HFH. Being observed in 526 patients (42.8%), elevated sPAP (>46 mm Hg) was associated with more severe heart failure symptoms at baseline and follow-up. Importantly, NYHA functional class significantly improved and TR severity was significantly reduced irrespective of PH. Conclusions PH is an important outcome predictor in patients undergoing T-TEER for relevant TR. In contrast to previous studies, no significant differences were observed for patients with precapillary and postcapillary PH in terms of survival free from HFH

Alzheimer-Demenz bei Menschen mit einem Down-Syndrom

Hintergrund : Menschen mit einem Down-Syndrom (MmDS) haben ein genetisch bedingt stark erhöhtes Risiko, an einer früh beginnenden Alzheimer-Demenz zu erkranken. In einer Interviewstudie mit Ärzt:innen, Patientenvertretungen sowie Mitarbeitenden in Wohn- und Arbeitseinrichtungen wurden Defizite im medizinischen Versorgungsprozess und Lösungsansätze erhoben. Methodik : Es wurden 14 teilstrukturierte Interviews durchgeführt und über eine qualitative Inhaltsanalyse ausgewertet. Ergebnisse : Fehlende Kenntnisse und Erfahrungen von Ärzt:innen im Umgang mit sowie in der medizinischen Versorgung von MmDS zeigten sich als zentrale Herausforderung. Zudem ist die Diagnostik der Demenz bei MmDS aufgrund verschiedener Ursachen (u. a. Fehlen geeigneter Diagnostikinstrumente in der Regelversorgung, fehlende zeitliche/finanzielle Ressourcen) erschwert. Zweifel an der Wirksamkeit von Antidementiva wurden geäußert sowie Hintergründe für einen erhöhten Einsatz sedierender Medikamente diskutiert. Eine aufmerksame Verhaltensbeobachtung und eine Einbindung von Betreuenden, eine regelmäßige Überprüfung und Reduktion von Multimedikation sowie der Einsatz alternativer Verhaltensmodifikationstechniken wurden als Lösungsansätze genannt. Schlussfolgerung : Die identifizierten Defizite in der medizinischen Versorgung der Zielpopulation sowie die Lösungsansätze gehen in die Entwicklung gesundheitspolitischer Handlungsempfehlungen ein, um die Versorgungssituation nachhaltig zu optimieren.Background : People with Down syndrome have a genetically increased risk of developing early onset Alzheimer’s dementia. An interview study with healthcare providers, patient representatives and employees in residential and work facilities was conducted to identify deficits in the healthcare process and approaches to overcoming them. Method : In this study 14 semi-structured interviews were conducted and analyzed using qualitative content analysis. Results : A lack of knowledge and experience on the part of medical service providers in dealing with and providing medical care for people with Down syndrome was identified as a key challenge. In addition, the diagnosis of dementia in people with Down syndrome is difficult for various reasons (including lack of appropriate diagnostic tools in standard care and lack of time or financial resources). Doubts were expressed about the efficacy of antidementia medications and the reasons for the increased use of sedatives were discussed. Attentive observation of behavior and involvement of caregivers, regular review and reduction of polypharmacy and the use of alternative behavior modification techniques were mentioned as possible solutions. Conclusion : The identified deficits in the medical care of the target population and the approaches to solving them will be incorporated into the development of health policy recommendations in order to optimize the care situation of those affected in the long term

Ethiopic Palimpsests

While spectacular scholarly breakthroughs and technological advances have led to a surge in recent scholarship on palimpsests, their existence within the Ethiopic manuscript tradition has hitherto received virtually no attention. Nonetheless, at least sixteen Ethiopic palimpsests have been identified, with many more to be found within collections inaccessible at present. Those available for study exhibit strong tendencies towards having biblical commentaries overwritten and being manufactured around the seventeenth century, though the only such manuscript to have undergone multi-spectral imaging thus far (Berlin, Staatsbibliothek zu Berlin, Petermann II Nachtrag 24) is of slightly later date. This manuscript contains no less than ten discrete scriptiones inferiores, but in this regard it likely constitutes an outlier to palimpsesting as practised within the Ethiopic tradition more broadly

Fixing sustainability through technoscience and diversity: The case of EU agriculture policy

Sustainability is a conveniently vague boundary term with which a variety of interest groups can identify. Over time, it has grown together with a technoscientific paradigm which demands a closer look at how actors envision science, technology, digitization, and innovation to foster said sustainability, and how the latter has shifted as a result. Sustainability also continues to hold strong value and political weight in the EU, where technoscientific optimism has had a binding effect, particularly in efforts of environmental protection in agriculture (in light of the Green Deal), in an increasingly decentralized political union. This paper discusses these processes in the recent reform of the Common Agriculture Policy (CAP; 2023–2027) by focusing on the one hand on how sustainability’s three pillars – the environmental, the social, and the economic – are ‘reconciled,’ and on the other, on the new ‘eco-schemes’ as an instrument to achieve a more sustainable agriculture. Empirical data gleaned from participant observation, expert interviews and policy document analysis show how in EU agriculture policy science, digitization/technology and innovation are imagined as fixtures that cohere these pillars, thereby maintaining a growth paradigm imminent to dominant sustainability discourses. This technoscientific sustainability is also evident in agriculture measures on the ground, in the new eco-schemes, which offer a diversity of farming approaches for EU's member states, ranging from agroforestry to precision farming. In this technocratic instrument, holistic systems, like agroecology, are rendered technical ‘tools’ that member states can combine at will, fostering a politics of toolkit diversity that accommodates diverse farming approaches and philosophies while evading environmental compliance. Scientific epistemology, technical quantification, digital tools and innovation thus act as wider discursive fixture that not only hold together the holy trinity of sustainability, but also accommodates diverse landscapes and member states, and through that the political union of an increasingly decentralized EU

Designing a Digital Flash Reading Test for Data-Based Decisions in Inclusive Classrooms: Duration and Word Length as Difficulty-Generating-Item Characteristics

Standardized assessment tools are essential for informed, data-driven decision-making. Reading speedily is a crucial early skill that all students should have the opportunity to develop in inclusive classrooms. To facilitate classroom-based reading diagnostics in this area of reading, we developed a flash reading test that reliably measures the performance of students with and without learning disabilities and intellectual disabilities. This test can be administered in the classroom and completed independently by students, taking only a few minutes, without requiring them to read aloud. The test is designed to provide an accurate assessment of the speed of lexical recall for all students. To evaluate the difficulty-generating-item characteristics of the new instrument, 400 primary and special school students participated in the test. The results indicate that students with low abilities and disabilities are particularly differentiated by the combination of a short display duration and short words. We provide information for test developers interested in designing similar assessments and teachers who can use this instrument to make informed decisions in the classroom