Background

Panallergens are ubiquitously expressed molecules that may drive IgE sensitization across diverse allergen sources, thereby shaping complex clinical phenotypes such as asthma and pollen–food allergy syndrome (PFAS). Although several panallergen families have been extensively characterized, unexplained sensitization patterns are still observed in clinical practice. Cyclophilins, a conserved protein family, have recently been proposed as candidate panallergens; however, their molecular, clinical, and functional roles remain largely undefined.

Methods

Within the Panallergens in Pediatrics (PAN‐PED) cohort, we investigated 100 Italian children with seasonal allergic rhinitis (SAR) for IgE sensitization to the cyclophilins Bet v 7 and Ara h 18. Functional assays were performed by sensitizing mast cells with patient sera ( n  = 11) and assessing activation upon stimulation with increasing concentrations of recombinant Bet v 7 (rBet v 7), using CD63 expression as readout. Additional experiments evaluated the impact of heat treatment (60°C, 80°C, or 100°C) and saliva digestion on rBet v 7 activity.

Results

rBet v 7 induced mast cell activation in a dose‐dependent manner, with peak responses at intermediate allergen concentrations, followed by attenuation at higher doses. rBet v 7 maintained functional activity after exposure to mild heat and saliva. A strong correlation was found between specific IgE levels to Bet v 7 and Ara h 18 ( r  = 0.996, p  < 0.0001).

Conclusion

Bet v 7 has the functional capacity to activate mast cells and is resistant to mild heat and saliva digestion, supporting its role as a clinically relevant panallergen. In addition, Bet v 7 shows high similarity to cyclophilins from other plant sources and exhibits virtually identical IgE‐binding properties to Ara h 18. These findings suggest a potential rationale for considering the inclusion of cyclophilins into molecular diagnostic panels, particularly in children with SAR.

Group‐living animals form dominance hierarchies to mediate competition among members. The social rank of individuals determines their access to resources such as food and mates, as well as their exposure and opportunities to respond to social challenges. High rank often translates into greater reproductive success, but the strength of this correlation may vary considerably depending on the rank metric and the reproductive trait studied. Here, we investigated the predictive ability of two rank metrics—ordinal and standardised ranks—assumed to capture distinct underlying mechanisms of intragroup competition on six reproductive traits for female and male spotted hyenas Crocuta crocuta . We used 28 years of individual‐based data from 481 hyenas from the Ngorongoro Crater, Tanzania. Ordinal rank strongly outperformed standardised rank in its ability to predict four out of six traits. As ordinal rank reflects the absolute position of an individual in the hierarchy, its importance when predicting reproductive traits suggests the relevance of priority of access to resources for both sexes. One trait—interbirth interval—was instead best described by standardised rank, suggesting socially mediated constraints on reproductive output. Our results reveal mechanisms that have been previously overlooked in explaining rank‐related success in specific reproductive traits in the spotted hyena and emphasise the importance of selecting rank metrics that appropriately reflect the competitive context in focus.

To compensate for the high computational costs when modelling large-scale mountain glaciers, ice fields or ice sheets over multiple millennia, it is common practice to coarsen the spatial resolution of numerical models to 1 km or more, which is not sufficient to describe complex valley topographies. Here, we examine the influence of spatial resolution by modelling a growing and retreating exemplary ice field in the European Alps at resolutions ranging from 50 m to 2 km using the Instructed Glacier Model (IGM). We find that while ice-covered areas remain similar, ice volume increases substantially with coarser resolution. Compared to the reference run at 50 m spatial resolution, model results at a resolution of 300 m and finer are comparable and sufficiently accurate to simulate topographically constrained ice flow. However, at resolutions coarser than ∼800m, topographic resampling artificially lowers slope angles and mountain peaks, providing a larger accumulation area at high altitudes, with thicker glaciers that are typically warm-based, while thinner glaciers at fine resolutions remain cold-based. Raised valley floors at coarse resolutions result in slower-flowing ice with increased thickness and glacial response times. The resulting hysteresis between climate forcing and glacial response at coarse resolutions is only partially decreased with slower temperature change. Seemingly stable model results at coarse resolutions may be misleading and accurate glacier geometries might arise from parameter choices that compensate for poorly resolved topography. Similar non-linear and altitudinal-dependent resolution effects are likely in mountain regions worldwide and emphasize the need for model advances to enable simulations at sufficiently high spatial resolutions to accurately resolve glacier dynamics.

Sialyl Lewis A (sLeA), or the CA 19-9 marker, is the best validated and only FDA-approved serologic marker clinically used to monitor recurrence, progression, and therapy efficiency in pancreatic ductal adenocarcinoma (PDAC) patients, making it an attractive target for antibody development. Recent clinical trials have demonstrated satisfying safety profiles and unique expression in a range of malignancies that highlight CA 19-9 as an attractive TACA to target using a stand-alone drug or an adjuvant therapy. Hence, we set out to explore the use of synthetic sLeA in the development of additional monoclonal antibodies (mAbs) with enhanced sLeA recognition and improved efficacy. Two mAbs targeting sLeA were generated through mice immunization with synthetic sLeA glycoconjugates, synthetic glycan arrays, and hybridoma technology. We then compared the antigen-binding properties of the newly developed mAbs with those of the widely used mAb 1116-NS-19-9 and demonstrated improved affinity and specificity for native sLeA ectopically expressed in B16 melanoma cells, surpassing the performance of the established mAb 1116-NS-19-9. Of the two mAbs, GB11 was more promising. Therefore, to elucidate the structural origin of improved GB11’s antigen binding, we conducted high-resolution mapping of the molecular recognition patterns between sLeA and the different antibodies using X-ray crystallography and STD NMR. These analyses revealed subtle yet critical differences in the glycan engagement and identified key structural features underlying enhanced GB11’s recognition of sLeA. MD simulations further supported these observations, indicating distinct orientations of sLeA within the binding pockets of each mAb. Our results suggest improved recognition of the native sLeA antigen by the newly generated GB11 antibody, providing a detailed and high-resolution elucidation of the molecular interactions underlying this recognition. Our study provides a tool with improved theranostic properties against sLeA-overexpressing malignancies.

Background

Subsyndromal generalized anxiety is highly prevalent and associated with impaired well-being, elevated stress, and functional limitations, yet affected individuals often do not meet criteria for guideline-based treatment. Scalable, low-threshold digital interventions that target psychophysiological regulation may help address this gap. Guided self-hypnosis and aromatherapy using essential oils have each demonstrated anxiolytic and relaxation-promoting effects. Combining these approaches may enhance efficacy and allow for conditioning of relaxation responses via olfactory cues.

Methods

This study protocol describes a randomized controlled trial evaluating the efficacy and conditioning potential of a digital self-hypnosis intervention combined with essential oil inhalation in adults with subsyndromal generalized anxiety. A total of N  = 630 participants will be randomized into six groups. Four groups enter the primary efficacy analysis: (1) self-hypnosis + bergamot essential oil, (2) self-hypnosis + lavender essential oil, (3) self-hypnosis without essential oil, and (4) a minimal-intervention control. The intervention is delivered online over six weeks (Phase 1), followed by a two-week conditioning phase without hypnosis (Phase 2), in which stimulus-specific effects of the essential oils are tested. The primary outcome is subjective relaxation, measured by the Multidimensional Mood Questionnaire (MDBF) at baseline, post-intervention (6 weeks), and post-conditioning (8 weeks). Secondary outcomes include anxiety symptoms, perceived stress, sleep quality, well-being, and worry. In a voluntary subsample, heart rate variability (HRV) and pulse wave variability (PWV) will be assessed as physiological correlates of relaxation. In addition, the questionnaires are expanded to include open-ended questions, enabling an exploratory assessment of participants’ experiences, attitudes, and reflections on the intervention and its potential for sustainability. This approach complements quantitative results with qualitative insights and may reveal new perspectives for future research.

Discussion

This study is expected to provide evidence on the efficacy of essential oil-enhanced digital hypnosis for subsyndromal anxiety and will examine whether repeated pairing of hypnosis and olfactory stimulation induces conditioned relaxation responses. If effective, this multimodal, low-intensity intervention could represent a scalable preventive approach for individuals with increased anxiety who are not receiving formal treatment and have been medically diagnosed.

Clinical Trial Registration: https://www.drks.de/search/de/trial/DRKS00039047/details , Identifier DRKS00039047.

This special issue aims to deepen the understanding of the Europeanisation of universities from 1980 to the mid-1990s. By focussing on five universities situated in different European contexts, we aim to provide insight into how Europeanisation has taken place and changed higher education. The geographical diversity and the main differences between universities provide the opportunity to show both differences and similarities in the process of European integration that the academic system underwent during a crucial period in contemporary history. In addition, we will relate the transformations of the universities to more general and overarching currents during this period. One of them is the acceleration of European integration in the years around 1990 when new areas became part of the political project. Another one is the emergence of the so-called knowledge society as a new political discourse and self-understanding. In this context, our comparative approach will be analytically rewarding, where we draw on our expertise of individual universities set in different societies and academic systems.

Childhood trauma (CT) is associated with cognitive impairment across major psychiatric disorders. We tested a novel transdiagnostic hypothesis that atypical connectivity of the default mode network (DMN) mediates the association between childhood trauma (CT) and cognitive impairment. The sample of 1851 individuals aged 18-25 included 433 patients with depression, eating disorders, alcohol use disorder, psychosis and ADHD) and were recruited as part of the ESTRA/STRATIFY/IMAGEN studies. CT was measured using the Childhood Trauma Questionnaire (CTQ). The CANTAB spatial working memory task was administered to assess cognition. Four a priori seeds of the default mode network (DMN) were measured during face processing, namely the medial prefrontal cortex (PFC), right lateral parietal (LP), left lateral parietal (LP) and posterior cingulate cortex (PCC), according to the Harvard-Oxford Cortical and Subcortical Atlas ( http://www.cma.mgh.harvard.edu/fsl_atlas.html ) as implemented in CONN. Patients had significantly reduced DMN connectivity between the four chosen DMN seeds and the rest of the brain. Reduced DMN connectivity mediated the association between higher CT and worse cognitive performance. Our findings are transdiagnostic in nature with stronger effects in some regions observed in depression, and suggest one transdiagnostic cortical network via which CT’s effects on cognition are transmitted.

Die Digitalisierung der Pflege kann nur dann wirksam und nachhaltig gelingen, wenn sie an den realen Versorgungs- und Arbeitsprozessen partizipativ ausgerichtet und konsequent kompetenzorientiert gestaltet wird. Pflegende sind als kompetente Anwender:innen, Gestalter:innen und Multiplikator:innen technologischer und digitaler Innovationen systematisch zu stärken und frühzeitig in Entwicklungs- und Implementierungsprozesse einzubinden. Das Cluster Zukunft der Pflege, gefördert durch das Bundesministerium für Forschung, Technik und Raumfahrt, fungiert hierbei als zentrale Innovations-, Transfer-, Beratungs- und Qualifizierungsplattform und unterstützt Individuen, Gesundheits- und Bildungseinrichtungen bei der Auswahl, Erprobung und nachhaltigen Implementierung innovativer Technologien für die Pflege- und Gesundheitsversorgung. Für Wissenschaft, Gesellschaft, Wirtschaft und Politik steht das Cluster Zukunft der Pflege als kompetenter Ansprechpartner für alle Fragen rund um das Thema digitale Transformation in der Pflege- und Gesundheitsversorgung zur Verfügung. Vorliegendes Positionspapier legt den Schwerpunkt auf die Kompetenzentwicklung im Kontext der Digitalisierung. Die zentralen Forderungen beziehen sich auf die standardisierte curriculare Einbindung digitaler Kompetenzen in Studium, Aus-, Fort- und Weiterbildung in der Pflege, die Bereitstellung innovativer Lernumgebungen und Schaffung kompetenzorientierter Rahmenbedingungen für eine praxistaugliche Anwendung technologischer Innovationen in der Pflege- und Gesundheitsversorgung sowie die Entwicklung und Etablierung neuer Kompetenzprofile im Kontext der digitalen Transformation.

While large language models (LLMs) such as ChatGPT are increasingly integrated into language education, little is known about their use for real-time spoken interaction in Chinese as a Foreign Language (CFL), particularly in formal K–12 instructional settings. This study investigates how 28 high school CFL learners in Austria engaged in tightly scripted question–answer interactions with ChatGPT’s Voice Mode during a classroom-based oral practice task. Data includes 28 audio recordings, six video recordings, and 26 written reflections collected immediately after the task. The analysis draws on a conversation-analytic perspective, focusing on task initiation, conversation openings, repair, and the role of teacher scaffolding, and is complemented by a thematic analysis of learner reflections. Findings show that ChatGPT (GPT-4) can be successfully constrained to deliver predictable, beginner-level interactions in Mandarin, offering learners a safe space to practice formulaic expressions and basic turn-taking routines, however, teacher support during task initiation and breakdowns was crucial to sustaining the interaction. Learners’ reflections revealed ambivalence: while many described the experience as motivating, novel, and low-anxiety, others expressed concerns regarding robotic prosody, occasional glitches, and the absence of interactional features typical of human interlocutors. We conclude that ChatGPT’s Voice Mode holds promise as a complementary tool for oral practice in beginner CFL classrooms, provided that its use is carefully mediated by teachers and supported by further AI development to enable seamless code-switching and multilingual feedback.

We investigate state designs for continuous-variable quantum systems using the aid of latticelike quantum states. These are code states of Gottesman-Kitaev-Preskill (GKP) codes. We show that the set of all GKP states forms a rigged continuous-variable state 2-design for an 𝑛-mode system. We use these lattice state designs to construct a continuous variable shadow tomography protocol, derive sample complexity bounds for both global and local GKP shadows under reasonable physical assumptions, and provide the physical gadgets needed to implement this protocol.

Biological invasions cause animal suffering, but few studies assess these welfare impacts, and hence understanding of them is limited. We present a framework which can be used to identify relative changes to the welfare of an individual animal caused by biological invasions. We use it to assess the welfare impacts of bird and ant invasions. These impacts are a global phenomenon affecting native and introduced animals. Nevertheless, some introduced ant species cause severe impacts wherever they occur, whereas introduced birds do not. Impacts are likely to be underreported, particularly those affecting introduced animals. Physical and behavioural evidence (e.g., injuries and repetitive preening by birds) is sufficient to identify many welfare impacts. Physiological evidence (e.g., changes in ‘stress’ hormones) is more scarce, and could provide useful additional information to help quantify impact severity. Published biodiversity impacts of biological invasions are an unmined resource that may be used to assess impacts.

Time-harmonic elastography (THE) is an emerging ultrasound imaging technique that allows full-field mapping of the stiffness of deep biological tissues. THE's unique ability to rapidly capture stiffness in multiple tissues has never been applied for imaging skeletal muscle. Therefore, we addressed the lack of data on temporal changes in skeletal muscle stiffness while simultaneously covering stiffness of different muscles. Acquiring repeated THE scans every five seconds we quantified shear-wave speed (SWS) as a marker of stiffness of the long head (LHB) and short head (SHB) of biceps brachii and of the brachialis muscle (B) in ten healthy volunteers. SWS was continuously acquired during a 3-min isometric preloading phase, a 3-min loading phase with different weights (4, 8, and 12 kg), and a 9-min postloading phase. In addition, we analyzed temporal SWS standard deviation (SD) as a marker of muscle contraction regulation. Our results (median [min, max]) showed both SWS at preloading (LHB: 1.04 [0.94, 1.12] m/s, SHB: 0.86 [0.78, 0.94] m/ s, B: 0.96 [0.87, 1.09] m/s, p <0.001) and the increase in SWS with loading weight to be muscle-specific (LHB: 0.010 during loading increased continuously over time by 0.022 [0.004, 0.051] m/s/min (p < 0.01). Using an exponential decay model, we found an average relaxation time of 27 seconds during postloading. Analogously, SWS SD at preloading was p <0.05) and increased by 0.005 [0.003, 0.008] m/s/kg (p <0.01) with loading. SWS SD did not change over loading time and decreased immediately in the postloading phase. Taken together, THE of skeletal muscle is a promising imaging technique for in vivo quantification of stiffness and stiffness changes in multiple muscle groups within seconds. Both the magnitude of stiffness changes and their temporal variation during isometric exercise may reflect the functional status of skeletal muscle and provide additional information to the morphological measures obtained by conventional imaging modalities.

Variational quantum algorithms (VQAs) represent a promising pathway toward achieving practical quantum advantage on near-term hardware. Despite this promise, for generic, expressive ansätze, their scalability is critically hindered by barren plateaus—regimes of exponentially vanishing gradients. We demonstrate that initializing a hardware-efficient, Floquet-structured ansatz within the many-body localized (MBL) phase mitigates barren plateaus and enhances algorithmic trainability. Through analysis of the inverse participation ratio, entanglement entropy, and a novel low-weight stabilizer Rényi entropy, we characterize a distinct MBL--thermalization transition. Below a critical kick strength, the circuit avoids forming a unitary 2-design, exhibits robust area-law entanglement, and maintains non-vanishing gradients. Leveraging this MBL regime facilitates the efficient variational preparation of ground states for several model Hamiltonians with significantly reduced computational resources. Crucially, experiments on a 127-qubit superconducting processor provide evidence for the preservation of trainable gradients in the MBL phase for a kicked Heisenberg chain, validating our approach on contemporary noisy hardware. Our findings position MBL-based initialization as a viable strategy for developing scalable VQAs and motivate broader integration of localization into quantum algorithm design.

The research proposal aimed to investigate the composition, architecture and function of mul-ti-protein/RNA complexes or degradosomes assembled by the RNA helicase UPF1 in path-ways of functional mRNA turnover, using a combination of biochemical, structural and cell-based methods. The decay of mRNA is a crucial step in post-transcriptional gene regulation as it ensures that the level and quality of gene expression in a cell is maintained at all times. Accordingly, cells have developed mechanisms to make the process of mRNA decay as effi-cient and robust as possible. One of the ways by which cells streamline mRNA decay is by assembling decay-competent RNA-protein complexes known as degradosomes. The minimal degradosome contains an RNA helicase and a ribonuclease, while more complex assemblies might include a polymerase and a “sensor” protein that signals for RNA decay. The helicase UPF1, best known for its function in nonsense-mediated mRNA decay (NMD), is also in-volved in several pathways that mediate turnover of functional cellular mRNA. While the mo-lecular mechanisms of UPF1 in context of an NMD-assembly are very well understood, com-paratively little is known about its recruitment and functionality in pathways of mRNA turnover. To this end, I proposed to investigate two UPF1-dependent mRNA turnover pathways – Stau-fen-mediated mRNA decay (SMD) and decay of replication-dependent histone mRNAs. The SMD pathway plays an important role in cellular development and differentiation, such as adi-pogenesis and neurogenesis, while histone mRNA decay at the end of the S-phase of the cell cycle ensures genome stability by coupling the levels of histone proteins to the amount of newly synthesized DNA. In these pathways, UPF1 associates with specific trans-acting pro-tein factors to assemble an mRNP that facilitates mRNA decay. These mRNPs can be con-sidered as UPF1-centric degradosomes and are the key functional unit of each pathway. Therefore, to understand the molecular mechanisms of these pathways, it is imperative to determine the composition of these mRNPs and investigate their stepwise assembly and dis-assembly as well as the regulation of UPF1 within these degradosomes. We used a combina-tion of structural (primarily cryo-electron microscopy) and biochemical methods, together with cell-based tools, to determine the molecular mechanisms of histone mRNA decay and identify overarching principles of function and regulation of UPF1-centric degradosomes. Together with our previous studies on SMD, this project provides invaluable insights into the regulation of UPF1 and the crosstalk among different UPF1-mediated mRNA degradation pathways.

Background: The 300IR house dust mite (HDM) sublingual immunotherapy (SLIT) tablet is approved for treatment of HDM-induced allergic rhinitis (AR). To provide a comprehensive review of the 300IR HDM-SLIT tablet safety pro file based on randomized controlled trial (RCT) pooled data and post-marketing (PM) pharmacovigilance data.

Methods: Subjects (5 -65 years) with con firmed HDM-AR with or without controlled asthma were treated with 300IR or placebo in 8 RCTs. Reported treatment-emergent adverse events (TEAEs) were pooled and analyzed descriptively in subsets of adults/adolescents and children. Adverse reactions (ADRs) collected from spontaneous reporting and PM studies through a pharmacovigilance system since the first marketing authorization were also analyzed.

Results: Across RCTs, 1853 subjects were treated with the 300IR HDM-SLIT tablet and 1846 with placebo. In both subsets of adults/adolescents and children whichever their asthma status, treatment-related TEAEs of higher incidence in active groups vs placebo were mostly consistent with mild or moderate local application-site reactions. They were mainly reported on the first days of treatment and decreased over time. 4 severe laryngopharyngeal reactions (2 requiring adrenaline/epinephrine) and 1 moderate eczema considered serious rapidly resolved with medications; no anaphylaxis was reported. In PM settings, ADRs reported in more than 235,000 patients were in line with RCT findings. Severe systemic reactions occurred rarely; 12 anaphylactic reactions resolved safely (5 with adrenaline). No new safety signal was raised.

Conclusion: Safety data from RCTs and more than 7 years of real-life experience con firmed the favorable safety pro file of 300IR HDM-SLIT tablet in patients across different regions, regardless of age and asthma status. Clinical trial registrations: NCT00674700; Retrospectively registered 06 May 2008. NCT01199133; Retrospectively registered 09 September 2010. NCT01527188; Retrospectively registered 01 February 2012. NCT02443805; Registered 29 April 2015/EudraCT 2014-004223-46; Registered 16 September 2015.

We present the rare case of a 51-year-old male diagnosed with a solitary fibrous tumor (SFT) of the prostate, along with a concurrent low-grade prostate adenocarcinoma (Gleason score 3 + 3, Grade Group 1). The diagnosis was confirmed by positive immunohistochemical markers, including CD34, BCL2, and STAT6, and molecular analysis showing a NAB2-STAT6 fusion. Following successful surgical management and the simultaneous diagnosis of a pulmonary relapse from a prior thyroid carcinoma, the patient remains under clinical surveillance. This is particularly significant given the patient's history of multiple tumors, including Hodgkin's lymphoma, papillary thyroid carcinoma, prostate cancer, and SFT.

Der Scope of Practice formuliert die Grundlagen und Standards der professionellen klinischen Praxis (nursing practice) für University Nurses (Bachelor Nurses) an der Charité - Universitätsmedizin Berlin. Es ist angelehnt an internationale Handlungsrahmen und dem Scope of Practice des International Council of Nurses. Beschrieben werden die rechtlichen Regelungen für Bachelor Nurses und die acht einzuhaltenden Standards in der nursing practice. Der erste Teil befasst sich mit der rechtlichen Grundlage für die professionelle klinische Praxis von Bachelor Nurses. Erläutert werden die Rahmenbedingungen und regulierten Titel, die den Handlungsspielraum von Bachelor Nurses bestimmen. Darüber hinaus dargestellt werden zentrale gesetzliche Regelwerke wie das Pflegeberufegesetz, die Ausbildungs- und Prüfungsverordnung und die Sozialgesetzbücher. Auf dieser Basis lässt sich der spezifische Tätigkeitsbereich der Bachelor Nurses an der Charité eindeutig umgrenzen. Der zweite Teil beschreibt acht Standards, die die professionelle klinische Praxis von Bachelor Nurses konkretisieren und in die praktische Anwendung überführen. Diese Standards definieren sowohl die Prinzipien der nursing practice als auch die Bedingungen und Grenzen ihrer Ausübung die bspw. für heilkundliche Tätigkeiten. Besondere Schwerpunkte liegen auf Verantwortung, Selbstreflexion, kritischem Denken und der Weiterentwicklung fachlicher Kompetenzen. Damit wird ein einheitlicher Rahmen geschaffen, um Bachelor Nurses im klinischen Alltag professionell zu verorten.

Primary human intestinal epithelial cells (IECs) require microenvironments that reproduce various in vivo cues to maintain survival, differentiation, and function in vitro . In this study, we investigated how intestinal stem cell (ISC)-derived monolayers respond to biomimetic substrates and shear stress using 3D-printed hydrogels based on bioactive decellularized and methacrylated small intestinal submucosa (dSIS-MA) integrated into a custom millifluidic system. Extensive bulk RNA sequencing experiments revealed that, compared with Matrigel-coated tissue culture plastic, dSIS-MA hydrogels supported survival- and differentiation-related signaling, stabilized gene expression over time, and promoted absorptive lineage maturation while reducing crypt-associated signatures. By applying dynamic culture conditions to the hydrogel system, IECs underwent transcriptional remodeling, characterized by activation of metabolic and immune pathways. Longitudinal analysis further indicated that shear stress enhanced metabolic pathway–associated gene expression and promoted differentiation toward absorptive lineages. These findings establish dSIS-MA hydrogels with controlled fluid flow as a biomimetic in vitro model that supports survival, maturation of human IECs and enables transcriptional adaptation to defined biochemical and mechanical cues, supporting future applications in disease modelling, drug testing, and regenerative medicine.

Synaptic neurotransmission is a critical hallmark of brain activity and one of the first processes affected in neural diseases. Monitoring this process, particularly synaptic vesicle recycling, in living cells has been instrumental in revealing the mechanisms responsible for neurotransmitter release. However, currently available reporters suffer from limitations, such as large probe sizes or limited compatibility for human neurons, hampering the quantitative analysis of synaptic pathophysiology. Here, we describe the NbLumSyt1 toolkit, a panel of nanobody-based affinity probes that target the luminal domain of the synaptic vesicle protein Synaptotagmin 1 (Syt1). These new tools enable quantitative, noninvasive imaging and functional interrogation of Syt1 exo-endocytosis and trafficking in human neurons, with unprecedented precision, versatility and cost efficiency, in technologies ranging from fixed- and live-cell super-resolution imaging to electron microscopy and mass spectrometry. Overall, NbLumSyt1 nanobinders provide a valuable platform for studying synaptic physiology and pathophysiology, benefiting fundamental neuroscience and translational efforts to study and develop treatments for brain-related disorders. Graphical Abstract