Neuromodulators control mood, arousal, and behavior by inducing synaptic plasticity via G-protein-coupled receptors. While long-term presynaptic potentiation requires structural changes, mechanisms enabling potentiation within minutes remain unclear. Using the Drosophila neuromuscular junction, we show that octopamine, the invertebrate analog of norepinephrine, potentiates evoked neurotransmitter release on the timescale of one minute via a G-protein-coupled pathway involving presynaptic OAMB receptors and phospholipase C. This fast potentiation correlates with elevated signals of the release factor Unc13A and the scaffolding protein Bruchpilot. Live, single-molecule imaging of endogenously tagged Unc13 revealed its instantly reduced motility and increased concentration in synaptic nanoclusters with potentiation. Presynaptic knockdown of Unc13A fully blocked fast potentiation. Moreover, deleting its N-terminal localization sequence mislocalized the protein fragment to the cytosol, but still allowed for rapid plasma membrane recruitment by diacylglycerol (DAG) analog phorbol esters and octopamine, implicating a role of more C-terminal domains. A point mutation of endogenous Unc13 in its DAG-binding C1 domain blocked plasticity-induced nanoscopic enrichment and synaptic potentiation. The mutation increased basal neurotransmission but reduced Unc13 levels, revealing a gain of function and potential homeostatic compensation. The mutation also blocked phorbol ester–induced potentiation, decreased the calcium sensitivity of neurotransmission, and caused short-term synaptic depression. Homeostatic potentiation induced by postsynaptic receptor block mirrored octopamine-induced Unc13 recruitment and required presynaptic OAMB receptors, indicating overlapping machinery. Thus, rapid Unc13 immobilization and nanoscale compaction are salient features of fast presynaptic potentiation.

Background

Family climate substantially influences children’s socio-emotional development. We examined mothers’ mental representations of their children and their relationships in three groups of mothers with young children (0–6 years): mothers (1) with a borderline personality disorder (BPD) (2), with a depressive or anxiety disorder but no BPD (AD/D), or (3) without a current mental disorder (CON). We expected both clinical groups to show more negative mental representations – more expressed emotion reflecting a critical attitude toward the child in general, more hostile attributions to child misbehavior in particular, and a less balanced view of the child (i.e., lower narrative coherence) – than CON mothers. We also expected mothers with BPD to have more impaired mental representations than mothers with AD/D.

Methods

Data were collected as part of an intervention study (at the pre-intervention assessment). To assess parental attributions, 172 mothers with BPD, 69 mothers with AD/D, and 96 CON mothers provided responses to vignettes and participated in a five-minute speech sample coded for expressed emotion and narrative coherence.

Results

BPD was associated with more criticism (OR = 3.17 and OR = 3.93) in comparison with CON mothers and mothers with AD/D, and with lower narrative coherence (OR = 5.45) compared with CON mothers but not compared with mothers with AD/D (OR = 1.41). Only narrative coherence remained significantly associated with group membership after education was controlled for. Mothers with BPD also showed more hostile attributions than CON mothers, with the AD/D group in between.

Conclusion

Without controlling for maternal education, critical attitudes toward the child in general were specifically associated with BPD, hostile attributions were less clearly associated, and narrative coherence was transdiagnostically associated with mental disorders in general. Once education was controlled for, disorder-specific associations were no longer observed, while transdiagnostic associations were maintained. Early interventions may specifically aim to decrease levels of criticism, help mothers increase non-hostile attributions of child misbehavior, and support mothers in building more coherent mental representations of their children.

Trial registration

This study was pre-registered at the German Registry of Clinical Studies (DRKS-ID: DRKS00020460).

Using Chinese panel data, we examine whether citizen well-being impacts the formation of political trust, which is key to regime stability. Through a quasi-experimental method, we demonstrate how an improvement in subjective well-being directly leads to increased political trust. In a supplementary analysis, we also demonstrate how low political trust is predictive of actions that undermine regime stability. These findings suggest that any government, even an authoritarian one, has an incentive to foster the happiness of its citizens.

Transfer RNA (tRNA) plays an essential role in protein translation, and tRNA modifications are important to their function. Recently, nanopore direct RNA sequencing (dRNA-seq) has shown promising results in the detection of complex tRNA modifications. However, its wider adoption in the tRNA field has been limited by a lack of (de)multiplexing solutions. Here, we present WarpDemuX-tRNA: an extension to the WarpDemuX method specifically optimized for multiplexed nanopore tRNA sequencing. Using consensus-based signal analysis using (soft) dynamic time warping and barycenter averaging, our approach improves barcode feature generation and achieves more robust barcode identification. WarpDemuX-tRNA outperforms the original method and achieves 99% precision and 95% recovery for four barcodes, while reducing computational complexity and runtime to 6 min per one million reads. WarpDemuX-tRNA is an open-source and free-to-use solution to high-throughput nanopore tRNA sequencing, facilitating more accessible, cost-effective, and high-throughput studies of tRNA modifications and their regulatory mechanisms.

We investigate the existence and uniqueness of solutions to first-order Stieltjes differential problems, focusing on the role of the Stieltjes derivative and its kernel. Unlike the classical case, the kernel of the Stieltjes derivative operator is nontrivial, leading to non-uniqueness issues in Cauchy problems. We characterize this kernel by providing necessary and sufficient conditions for a function to have a zero Stieltjes derivative. To address the implications of this nontrivial kernel, we introduce a function space which serves as a suitable framework for studying Stieltjes differential problems. We explore its topological structure and propose a metric that facilitates the formulation of existence and uniqueness results. Our findings demonstrate that solutions to first-order Stieltjes differential equations are, in general, not unique, underscoring the need for a refined analytical approach to such problems.

Here, we present the first fully varve-based chronology for the deposits of the deep-water Lake Shira (Chulym-Yenisei Basin, South Siberia), derived from a new sediment core. The very well-preserved varves show typical properties of the clastic-biogenic and endogenic types that can be subdivided into four sublayers representing winter–early spring, late spring, early summer and late summer–autumn. The analysed sediment section of 147 cm length comprises 2491 varve years with a total counting error of 1.6 % (i.e. ±40 years), making the new sediment core from Lake Shira a unique high-resolution archive for multi-proxy studies of past climate/environmental change and human-environment interactions. Direct comparison of nine AMS radiocarbon (14C) dates based on sediment bulk organic fractions with an age-depth model based on varve counting made it possible to examine the 14C reservoir effect in the lake. The reservoir effect is a common issue when estimating the age of environmental proxies from lacustrine sedimentary archives in Inner Asia. Although a constant reservoir effect is commonly used to correct the 14C dates from a single core or lake basin, our results from Lake Shira demonstrate that it varies significantly over the last 2500 years, ranging from 240 ± 30 to 1045 ± 30 years. The spatiotemporal variability of the reservoir effect can considerably reduce the accuracy of age-depth models based solely on the bulk organic sediment fraction. Where varved sediment is unavailable, as is usually the case, lignin phenols, terrestrial plant remains and purified pollen concentrates should be considered as alternative dating materials.

Oesophagostomum spp. and Ascaris suum represent the most common porcine nematodes and anthelmintic resistance (AR) to various anthelmintics has been reported for Oesophagostomum. However, the current AR status for worm populations on German farms and practical methods facilitating reliable AR detection are missing. Herein, the efficacy of benzimidazoles (BZ) (fenbendazole, 5 mg/kg body weight, single dose) was analysed on 13 farms with outdoor access. The Faecal Egg Count Reduction Test (FECRT) estimates for strongyles on the farms (range 99.8–100 %) exceeded the target efficacy (99 %) of the new W.A.A.V.P. guideline for Oesophagostomum dentatum. Deep amplicon sequencing was used for the first time for porcine nematodes and revealed no polymorphisms associated with BZ-resistance in codons 134, 167, 198 and 200 of the isotype-1 β-tubulin gene. Nemabiome analysis using ITS-2 deep amplicon sequencing, based on two pre- and post-treatment samples, showed a significant increase (p < 0.001) of Oesophagostomum quadrispinulatum after BZ treatment. For A. suum, the interpretation of FECRT estimates can be hindered due to coprophagy-associated false-positive egg counts in pigs. Therefore, two FECRT analysis for A. suum were pursued, the first analyses included all EPG data, the second considered EPGs <200 pre- and post-treatment as negative. An in ovo larval development assay (LDA) was developed for the in vitro analysis of BZ-susceptibility in A. suum. Computed EC50 values ranged from 1.50 to 3.36 μM thiabendazole (mean 2.24 μM). An EC50 of 3.90 μM thiabendazole (mean EC50 + 3 × SD) as provisional cut-off for detection of resistant populations is suggested. In conclusion, no AR was detected in Oesophagostomum using the FECRT and β-tubulin deep amplicon sequencing. For A. suum the FECRT results were ambiguous, in some cases even when excluding the low egg counts from calculations. With the in ovo LDA all investigated A. suum populations were identified as susceptible to BZ.

Metal-metal bonds between metal cations are ubiquitous in coordination complexes, whereas similar bonding characteristics between non-metal and metal cations are not. Here, we report an X-ray crystal structure of a centrosymmetric complex [Ag(m-O3SCF3)2{(4MePyNO)2I}]2 (1-AgI), where the iodine(I) cation forms an unusual 3-centre 4-electron [O–I–O]+ halogen bond with two oxygen atoms from two 4-methylpyridine N-oxide (4MePyNO). The iodine(I) atoms from two [4MePyNO−I−ONPyMe4]+ cations, which, together with the two silver(I) atoms from the paddlewheel [Ag(m-O3SCF3)2]22− structure, establish two unique I+–Ag+ bonds at 2.863(2) Å. This bond length is characteristic of bimetallic coordination bonds such as Au+−Ag+ [2.8553(6) Å], Cu+−Ag+ [2.8616(9) Å], and Pt2+−Ag+ [2.8602(4) Å]. Density Functional Theory calculations unambiguously support the existence of the I+–Ag+ coordination bond between non-metal and metal cations in complex 1-AgI.

Clathrin-mediated endocytosis depends on complex protein interactions. Eps15 plays a key role through interactions of its three EH domains with Asn-Pro-Phe (NPF) motifs in intrinsically disordered regions (IDRs) of other endocytic proteins. Using nuclear magnetic resonance spectroscopy, we investigate the interaction between Eps15’s EH domains and a highly disordered Dab2 fragment (Dab2320-495). We find that the EH domains exhibit binding promiscuity, recognizing not only the NPF motif of Dab2 but also other phenylalanine containing motifs. This promiscuity enables interactions with Eps15’s own IDR (Eps15IDR), which lacks NPF motifs, suggesting a self-inhibitory state that promotes liquid-liquid phase separation. Despite competing for the same EH domain binding sites, Eps15IDR and Dab2320-495 can bind EH123 simultaneously, forming a highly dynamic interaction network that facilitates the recruitment of Dab2320-495 into Eps15 condensates. Our findings provide molecular insights into the competitive interactions shaping the early stages of clathrin-mediated endocytosis.

Encoding and manipulating digital information in quantum degrees of freedom is one of the major challenges of today’s science and technology. The valley indices of excitons in transition metal dichalcogenides (TMDs) are well-suited to addressing this challenge. Here, we employ mechanical strain to manipulate intervalley interactions and tune the valley polarization dynamics of excitons across a broader range of momentum space in monolayer TMDs. We use strain engineering to form valley-hybridized excitons that combine the advantages of bright intravalley excitons, where the valley index directly couples to light polarization, and dark intervalley excitons, characterized by low depolarization rates. We demonstrate that these valley-hybridized excitons exhibit signatures of coherently coupled states with a 100-fold reduction in valley depolarization rate and up to a 5-fold increase in steady-state valley polarization compared to previously studied excitons. Our findings of strain-tunable valley character of excitons advance the applications of TMDs in valleytronics.

Sleep is crucial for cognitive functions and life span across species. While sleep homeostasis and cognitive processes are linked through cellular and synaptic plasticity, the signaling pathways connecting them remain unclear. Here, we show that Drosophila insomniac (inc) short sleep mutants, which lack an adaptor protein for the autism-associated Cullin-3 ubiquitin ligase, exhibited enhanced Pavlovian aversive olfactory learning and memory, unlike other sleep mutants with normal or reduced memory. Through a genetic modifier screen, we found that a mild reduction of Protein Kinase A (PKA) signaling specifically rescued the sleep and longevity phenotypes of inc mutants. However, this reduction further increased their excessive memory and mushroom body overgrowth. Since inc mutants displayed higher PKA signaling, we propose that inc loss-of-function suppresses sleep via increased PKA activity, which also constrains the excessive memory of inc mutants. Our data identify a signaling cascade for balancing sleep and memory functions, and provide a plausible explanation for the sleep phenotypes of inc mutants, suggesting that memory hyperfunction can provoke sleep deficits.

Background

Human herpesvirus 6A (HHV-6A) is a member of the betaherpesvirus family and is associated with neurotropic diseases. Despite its clinical significance, HHV-6A research has been hampered by challenges in the generation of recombinant viruses. Although bacterial artificial chromosome (BAC) systems and well-established mutagenesis techniques are available for HHV-6A, its tendency towards latency and slow viral replication pose inherent challenges to reconstituting infectious virus. Virus reconstitution has been achieved by only a few laboratories worldwide and remains a hurdle for HHV-6A research.

Methods

We addressed these key bottlenecks of HHV-6A reconstitution by systematically refining nucleofection and stimulation conditions. Using a reporter virus, we improved cell preparation, implemented a dimethyl sulfoxide (DMSO) treatment, removed contaminating DNA by exonuclease V digestion, optimized the cell recovery after nucleofection, and assessed novel stimulation strategies that accelerate virus replication.

Results

In this study, we established a cost-effective and robust method for HHV-6A reconstitution. Combining DMSO and Exonuclease V pretreatment with an optimized recovery after nucleofection resulted in an increased transfection efficiency of up to 30%. Selected stimuli promoted lytic replication and facilitated the recovery of infectious virus. Combining IOX2 + hydrocortisone exceeded all other stimuli, reducing the reconstitution time to two weeks. Our optimized protocol has proven to be highly reproducible across multiple laboratories, different mutant viruses, instruments, and operators, ensuring reliability and broad applicability. It also allowed us to generate a novel reporter virus that shed light on the replication kinetics of the virus.

Discussion

This efficient HHV-6A reconstitution protocol addresses long-standing challenges, offering a widely adoptable method that simplifies recombinant virus generation and enhances future research into viral gene functions and infection mechanisms.

Background and aim

Soil health is vital for the sustainability of ecosystem services such as food and fiber production, nutrient cycling and water supply. Soil health can be assessed through a combination of physical, biological and chemical metrics. There is emerging evidence that reactive silica is a strong factor controlling soil functions.

Scope

We explain how reactive silica, specifically dissolved silicic acid and amorphous silica, can be used as a new metric for assessing soil health, complementing traditional metrics or even substituting for them.

Conclusion

The pivotal role of reactive silica for soil health is particularly important under stress conditions that are typically associated with drought and soil degradation. The status of reactive silica indicates soil degradation earlier than the currently used metrics, because reactive silica depletion is followed by soil degradation. We recommend suitable methods and benchmarks for assessing reactive silica. Furthermore, we suggest further research to improve our understanding of the importance of reactive silica for soil health. We call upon the soil research community to include reactive silica as a metric for soil health assessment.

The increasing antimicrobial resistance (AMR) in Staphylococcus pseudintermedius causing skin and soft-tissue infections (SSTIs) in companion animals is a public health concern. The aim of this study was to verify if mobile genetic elements (MGEs), in particular plasmids, are related to the carriage of AMR genes among circulating and clinically relevant S. pseudintermedius. In total, 56 S. pseudintermedius, representing predominant and emerging clonal lineages associated with SSTIs in dogs and cats collected in Lisbon (Portugal), were subjected to plasmid DNA extraction and digestion with EcoRI and XbaI. Each unique restriction pattern was assigned to a plasmid profile. A subset of 17 strains was further selected for hybrid whole genome sequencing (WGS) on Oxford Nanopore MinION and Illumina MiSeq platforms. Thirty-one of the 56 S. pseudintermedius strains carried one or more plasmid(s), mostly of small or medium sizes, corresponding to eight plasmid profiles. Two of the identified plasmids carried AMR determinants; plasmid pSP-G3C4, isolated from ST71 strains, carried the tetracycline resistance gene tet(K) and plasmid pSP5912, isolated from a ST2061 strain, harbored the qacG biocide resistance gene. Other AMR determinants were detected as part of MGEs integrated into the bacterial chromosomal DNA, namely Tn552, Tn552-like, Tn553, Tn916, Tn5405-like, Tn5801, Tn5801-like GI6287 and pRE25-like elements. In addition, a new chromosomal cassette, carrying fusC, was identified in a ST1183 strain. The 12 methicillin-resistant S. pseudintermedius studied carried staphylococcal cassette chromosome mec (SCCmec) type III (n = 5), SCCmec type IVg (n = 3), SCCmecNA45 (n = 1), ΨSCCmec57395 (n = 1), the recently described cassettes SCCmec7017–61515 (n = 1), or SCCmec type V(T)SL/154 (n = 1). Most strains carried intact prophages without AMR determinants. Intact restriction-modification systems were detected in 12 out of the 17 strains and CRISPR/Cas in five strains, four of which were methicillin-susceptible. The results of this study suggest that the AMR content in S. pseudintermedius is mainly related to MGEs integrated into the chromosomal DNA rather than located on plasmids. These results provide important insights that may lead to a better understanding of multidrug resistance in S. pseudintermedius towards improved SSTIs treatment in companion animals.

Introduction

Mental disorders are among the leading causes of the global burden of disease and are often associated with severe functional impairment and high societal costs. Psychotherapeutic, psychopharmacological and internet-based mental health interventions have proven to be helpful, but challenges remain, including only moderate response rates, high relapse rates and barriers to accessing mental healthcare. Much of the existing evidence stems from studies conducted in controlled, often standardised settings that only partially reflect real-world conditions, contributing to a ‘scientist-practitioner gap’. Moreover, the mechanisms of change, such as the interaction between treatment intensity, common factors (eg, the therapeutic relationship) and specific intervention techniques, have not been sufficiently investigated. In particular, the relationship of changes in personality functioning (PF) with mental and physical health has not yet been extensively researched.

Methods and analysis

The PSYMPACT (Psychological Impact Factors of Mental Health Treatments) study will use a longitudinal study design with a naturalistic sample (N ≈ 3000) to examine changes in psychopathology, PF and allostatic load in psychotherapeutic, psychopharmacological and internet-based treatments. The aim is to identify factors contributing to improvements and deteriorations in mental and physical health across different settings, including common and specific factors. Additionally, to provide patient perspectives, qualitative interviews will be conducted with individuals with varying levels of severity of mental health problems. Allostatic load will be assessed using repeated hair cortisol measurements. Furthermore, ecological momentary assessment will be used to examine the diurnal variability of PF as well as its more momentary correlates and longer-term outcomes. The central research questions and aims include (1) the assessment of common factors across different treatment settings, (2) associations of specific and common factors with improvements in mental health, including PF, (3 and 4) the importance of treatment intensity and interaction effects with common and specific factors, (5) the association of changes in psychopathology with changes in allostatic load, (6) the trait and state variability of PF, (7) the identification of patients who deteriorate under specific treatments and (8) patients’ perspectives on the effectiveness of different treatment modalities.

Ethics and dissemination

Approval was obtained from the Ethics Committee of the Department of Education and Psychology at the Freie Universität Berlin, Germany. Results will be submitted to peer-reviewed specialised journals and presented at national and international conferences.

Registration details

Before data collection started in November 2024, the study was registered in the German Clinical Trials Register (https://www.drks.de/search/de/trial/DRKS00035560).

Cognitive behavioral therapy (CBT) is an effective treatment for obsessive-compulsive disorder (OCD). However, CBT does not lead to a satisfying symptom reduction in a considerable number of patients with OCD. The identification of variables that predict insufficient treatment response could improve efficient treatment selection and inform the development of specific augmentative treatments. In the current study, we tested whether prediction of treatment response can be improved by including neurobiological markers during working memory (WM) performance. Forty-four patients with a primary OCD diagnosis participated in an n-back WM task with varying WM load while functional Magnetic Resonance Imaging (fMRI) was performed. Subsequently, all patients received CBT in an outpatient clinic. WM load-dependent modulation of the blood-oxygen-level-dependent (BOLD) signal in a bilateral cluster in inferior/superior parietal lobule predicted CBT response over and above clinical and sociodemographic variables (p < 0.05). Higher modulation was associated with larger relative symptom reduction. The results of the current study indicate that the ability of the WM system to flexibly adapt to changing task demands might be a useful indicator of CBT response in OCD. Possibly, this mechanism facilitates relearning processes during exposure-based CBT. However, findings need to be replicated in larger samples.

As climate change escalates, the Berlin-Brandenburg region faces new challenges. Climate change-induced extreme events including droughts, heatwaves, and floods, are expected to cause new conflicts to emerge and aggravate existing ones. To guide future research, we engaged a transdisciplinary academic community of experts to co-develop a list of key questions on these climate and water challenges in the region. Our findings highlight the urgent need for integrated and participatory research approaches. We expect this list of key questions to provide a roadmap for scientists and policymakers to foster actionable knowledge production to address climate and water challenges in the region.

The synthesis and photovoltaic effect of Cu3xBi1−xI3 (CBI) thin films were studied. Although CBI was originally inspired by the material class of Caswellsilverites (also known as Rudorffites), an XRD study showed that its crystal structure resembles a BiI3-cage with interstitial copper. CBI was found to demonstrate optoelectronic properties suitable for solar light harvesting, including a band gap of 1.66 eV and thermal stability up to 333 °C. The corresponding thin film solar cells achieved a peak power conversion efficiency of 0.65%.