eLife assessment

This important work has substantially advanced our understanding of the molecular basis of symmetry breaking and lineage specification in preimplantation mammalian embryos. The results generated using live imaging are compelling. Quantification of the functional assays is convincing and would be improved by increasing the number of embryos in the evaluations and clearly stating how many embryos are evaluated per experiment.

eLife assessment

This important study explores the impact of pH changes and cancer mutations on nucleosome interactions and higher-order chromatin structures. The evidence supporting the main conclusions is solid, based on rigorous computational methods, including pKa prediction, electrostatic force calculation, and molecular dynamics simulations. The findings provide insights into how protonation states and cancer-associated mutations affect nucleosome electrostatics and chromatin organization, making this work of broad interest to chromatin biologists, cancer researchers, and computational biophysicists.

eLife assessment

This valuable study investigates how inter-organ communication between the tracheal stem cells and the fat body plays a key role in the directed migration of tracheal stem cells in Drosophila pupae. While the experimental data are extensive and complementary, the evidence presented to substantiate some of the conclusions appears incomplete and requires further clarification and additional experiments. The work would be of interest to researchers in the fields of developmental biology and cancer biology.

eLife assessment

This study presents valuable findings on the relative cerebral blood volume of non-human primates that move us closer to uncovering the functional and architectonic principles that govern the interplay between neuronal and vascular networks. The evidence of areal variations is solid, but that of vessel counting and laminar analysis is incomplete. The lack of a direct comparison of their approach against better-established MRI-based methods for measuring hemodynamics and vascular structure weakens the evidence provided in the current paper version. The work will be of interest to NHP imaging scientists.

eLife assessment

In this elegant and thorough study, Sánchez-León et al. investigate the effects of tDCS on the firing of single cerebellar neurons in awake and anesthetized mice. They find heterogeneous responses depending on the orientation of the recorded Purkinje cell. The paper is important in that it may well explain part of the controversial and ambiguous outcomes of various clinical trials. It is a well-written paper on a deeply analyzed dataset and the methods in the paper are generally convincing, with the current version having some weaknesses in statistical reporting and power.

eLife assessment

This work presents valuable findings of a modulatory effect of yohimbine, an alpha2-adrenergic antagonist that raises noradrenaline levels, on the reconsolidation of emotionally neutral word-picture pairs, depending on the hippocampal and cortical reactivation during retrieval. The evidence supporting the conclusion is incomplete so far, particularly considering concerns about the median-splitting approach for reaction times and hippocampal activity. The work will be of broad interest to researchers working on memory.

eLife assessment

This study provides valuable evidence on the relationship between morphine-induced social deficits, corticotropin-releasing factor receptors, and alterations in neuronal activity in the paraventricular nucleus of the hypothalamus of mice (PVN). Convincing approaches and methods were used to show that the CRF1 receptor plays a role in sociability deficits occurring after acute morphine administration. Conclusions regarding mechanistic connections between the effect of modulation of CRF 1 receptor on sociability and PVN neuronal firing are, however, incompletely supported.

eLife assessment

This study combines experimental and theoretical approaches to examine metabolites at the single-cell level in tea plants. The authors skilfully integrated various tools available for this type of research, and meticulously presented and illustrated every step of the survey. The overall quality of the work is convincing, and it represents an important contribution to our understanding of the compartmentalization of biosynthesis pathways.

eLife assessment

This useful study provides data suggesting that subcellular localization of the spatial regulator of cell division, MinD, is an intrinsic feature of the protein's ability to associate with the membrane as both a dimer and a monomer. These findings distinguish the behavior of MinD in B. subtilis from its counterpart in E. coli and suggest that there is not a need to invoke additional localization factors. However, all three reviewers agreed that the study is incomplete: experimentally, quantitation and assessment of MinD behavior in the presence of proteins previously implicated in its localization are missing, among other assays, and the molecular modeling necessary to support the authors' conclusion that their data support a reaction-diffusion model is completely absent. Finally, the manuscript itself is difficult to read with an overly long discussion and disorganized introduction and results sections, and it will require significant revision.

eLife assessment

This important work by Zheng and colleagues uses a large cohort database from Shanghai to identify that post-infection vaccination among previously vaccinated individuals provides significant low to moderate protection against re-infection. The evidence supporting the conclusion is convincing with some limitations, e.g., lack of symptom severity as an outcome, and no inclusion of time since infection as an independent variable). This study will be of interest to vaccinologists, public health officials and clinicians.

eLife assessment

This study presents valuable evidence concerning the potential for naturalistic movie-viewing fMRI experiments to reveal some features that are correlated with the functional and topographical organization of the developing visual system in awake infants and toddlers. The data are compelling given the difficulty of studying this population, the methodology is original and validated, and the evidence supporting the conclusions is convincing and in line with prior research using resting-state and awake task-based fMRI. This study will be of interest to cognitive neuroscientists and developmental psychologists, and in particular those interested in using fMRI to investigate brain organisation in pediatric and clinical populations with limited tolerance to fMRI.

eLife assessment

In this important study, Bu et al investigate how cell overcrowding triggers a mechano-transduction pathway involving TRPV4 channels, focusing on high-grade ductal carcinoma in situ (DCIS) cells. The authors show that cell crowding in these malignant cells leads to a reduction in cell volume and promotes a pro-invasive phenotype through calcium homeostasis and TRPV4 channel trafficking to the plasma membrane; this phenomenon is specific to invasive cell lines like MCF10CA and DCIS and is corroborated by patient tissue samples. The work suggests the role of TRPV4 in cell motility and mechanical sensing, offering potential therapeutic insights for targeting cancer metastasis. While the study presents robust and convincing data, the absence of TRPV4 genetic ablation is a critical limitation, which would further confirm its role in these processes.

eLife assessment

This study provides novel insights into COVID-19 immune responses by using the delta of the normalised accessible surface area (DASA) to map IgM responses to the SARS-CoV-2 Membrane protein M1-subtype across multiple European cohorts. The evidence supporting the findings is solid, with thorough validation and comprehensive analysis, although additional clarity on T-independent B cell reactions and the impact of comorbidities would further strengthen the conclusions. The methods and data presented are valuable for advancing diagnostic and prognostic tools for COVID-19, particularly in the context of long COVID.

eLife assessment

This useful study characterises motor and somatosensory cortex neural activity during naturalistic eating and drinking tongue movement in nonhuman primates. The data, which include both electrophysiology and nerve block manipulations, will be of value to neuroscientists and neural engineers interested in tongue use. However, data analysis needs to be improved to strengthen the inadequate support for some of the main claims in the paper.

eLife assessment

The authors combined molecular dynamics simulations and experiments to study the role of ATP as a hydrotrope of protein aggregates. The topic is of major current interest and thus the study potentially makes an important contribution to the community. With the revised version, the level of evidence is considered generally solid, although there remains concern regarding the unusually high ATP concentration used in the simulation.

eLife assessment

The authors developed a method to allow a hypothermic agent, neurotensin, to cross the blood-brain barrier so it could potentially protect the brain from seizures and the adverse effects of seizures. The work is important because it is known that cooling the brain can protect it but developing a therapeutic approach based on that knowledge has not been done. The paper is well presented and the data are convincing. Revisions to clarify some of the methods and results improved the paper and more data about tolerability would improve the paper further.

eLife assessment

In this manuscript, the authors present valuable findings on the apparent role of a salience-network anterior insula node in directing fronto-parietal and default-mode network activity within a tripartite network during control of memory, drawn from an impressive invasive human neurophysiological dataset. We commend the use of a large intracranial EEG dataset to approach this question. While many aspects of the evidence are solid, there are some remaining points which are methodologically incomplete and could be further improved with some specific control analyses and word changes.

eLife assessment

This study provides compelling data that defines the structure of the S. cerevisiae APC/C. The structure reveals overall conservation of its mechanism of action compared to the human APC/C but some important differences that indicate that activation by co-activator binding and phosphorylation are not identical to the human APC/C. Thus this study will be of considerable value to the field.

eLife assessment

In this useful study, the authors investigate the regulatory mechanisms related to toxin production and pathogenicity in Aspergillus flavus. Their observations indicate that the SntB protein regulates morphogenesis, aflatoxin biosynthesis, and the oxidative stress response. The data supporting the conclusions are compelling and contribute significantly the advancing the understanding of SntB function.

eLife assessment

The regulation of mitosis and the dynamics of the mitotic spindle in it are central to cell division with high fidelity and crucial for normal division and development and defects therein can lead to disease. A key component of ensuring the fidelity is the "spindle assembly checkpoint". This valuable study using convincing experimental approaches in fission yeast has revealed novel links between the MAP-kinase signalling pathway modulating the spindle assembly checkpoint.

eLife assessment

This valuable study characterises receptors for calcitonin-related peptides from a deuterostomian animal, the echinoderm Apostichopus japonicus, by a combination of heterologous expression, pharmacological experiments, and the quantification of gene-expression levels. The authors provide solid evidence for a functional calcitonin-related peptide system in the sea cucumber, but some of the phylogenetic and statistical analyses and the evidence of a physiological function are incomplete. This work should be of interest to scientists studying the signaling pathways, functions, and evolution of neuropeptides, and could be of relevance to improving the culture conditions of this economically key species.

eLife assessment

This valuable paper uses the ChEC-seq2 technique to investigate RNA polymerase II (RNAPII) binding patterns in yeast and demonstrates that this technique is a complementary method for investigating transcription mechanisms, especially slow steps at the initiation and termination regions. The authors use ChEC-seq2 data to investigate RNAPII kinetics and obtain solid data providing new insights into transcription regulation. This study highlights the importance of careful methodological comparisons in understanding RNAPII dynamics.

eLife assessment

The manuscript by Li and coworkers analyzed astrocytic differentiation of midbrain floor plate-patterned neural cells originating from human iPS cells, with a LMX1A reporter. This valuable work identifies transcriptomic differences at the single-cell level, between astrocytes generated from LMX1A reporter positive or negative cells, as well as non-patterned astrocytes and neurons. The evidence is solid, but the paper can be strengthened by further analyses of the transcriptomic data, and astrocytic morphology; also, searching for some of the differentially expressed genes by immunohistochemistry in different regions of the mammalian brain, or in human specimens, would be very informative.

eLife assessment

In this valuable study, García-Vázquez et al. provide solid evidence suggesting that G2 and S phases expressed protein 1 (GTSE1), is a previously unappreciated non-pocket substrate of cyclin D1-CDK4/6 kinases. To this end, this study holds a promise to significantly contribute to an improved understanding of the mechanisms underpinning cell cycle progression. Notwithstanding these clear strengths of the article, it was thought that the study may benefit from establishing the precise role of cyclin D1-CDK4/6 kinase-dependent GTSE1 phosphorylation in the context of cell cycle progression, obtaining more direct evidence that cyclin D1-CDK4/6 kinase phosphorylate indicated sites on GTSE1 (e.g., S454) and mapping a degron in GTSE1 whose function may be blocked by cyclin D1-CDK4/6 kinase-dependent phosphorylation.

eLife assessment

This study presents a valuable optimization algorithm to identify polymer models that best fit population-averaged chromosome contact data that will be of interest to physicists and biologists working on chromatin organization. The conclusions are supported by solid evidence.

eLife assessment

In this valuable study, the authors studied a novel Zn2+- and NAD+-independent KDAC protein, AhCobQ, in Aeromonas hydrophila, which lacks homology with eukaryotic counterparts, thus underscoring its unique evolutionary trajectory within the bacterial domain. They attempt to demonstrate deacetylase activity, however, assays to detect this are still incomplete and require further refinement. The work will be of interest to microbiologists studying metabolism and post-translational modifications.

eLife assessment

This valuable study has characterized the unique expression of Schlemm's canal endothelial cells (SECs) using FACS-sorted specific cell bulk RNA-Seq and scRNA-/snRNA-Seq of mouse SECs. The compelling study identified novel biomarkers for SECs and molecular markers for two inner wall SEC states and outwall SECs in mouse eyes. Significant gene networks and pathways were elucidated for their potential contribution to glaucoma pathogenesis, providing targets for further research in relation to glaucoma.

eLife assessment

This important study introduces and evaluates the efficacy of a novel form of non-invasive brain stimulation in humans: kilohertz transcranial magnetic perturbation (kTMP). The evidence provided for the ability of kTMP to increase cortical excitability with minimal sensation is compelling, with two separate replication experiments. Although exploratory in nature, this work represents new avenues for non-invasive brain stimulation research that has potential long-term appeal for both clinical and research applications. This paper will be of significant interest to neuroscientists interested in brain stimulation.

eLife assessment

This study provides valuable insights and allows for hypothesis generation around diet-microbe-host interactions in alcohol use disorder. The strength of the evidence is convincing: the work is done in a rigorous manner in a well-described cohort of patients with AUD before and after withdrawal. There are several weaknesses, including validating the metabolites identified by metabolomics, the cross-sectional study design, the lack of a healthy control group, and the descriptive nature of such clinical cohort studies. Nevertheless, the study provides a wealth of new data that may be the basis for future studies that test causality and elucidate the role of single metabolites in the psychiatric sequela of AUD.

eLife assessment

This important study describes changes in excitability in motor neurons of the peripheral autonomous nervous system during aging. The manuscript provides convincing evidence indicating that sympathetic neurons from aged mice show higher excitability compared to neurons from young mice which was linked to decreased activity of KCNQ2/3 potassium channels. This research has implications for understanding the age-related changes that occur in the peripheral nervous system.

eLife assessment

This study presents useful findings for how sevoflurane anesthesia modulates the activity of corticotropin-releasing hormone neurons in the paraventricular nucleus of the hypothalamus and how manipulation of such PVHCRH neurons influences anesthesia and post-anesthesia responses. The technical approaches are solid and the data presented is largely clear. Whether PVHCRH neurons are critical for the mechanisms of sevoflurane anesthesia is a direction for the future.

eLife assessment

This valuable study explores a new strategy of lysin-derived antimicrobial peptide-primed screening to find peptidoglycan hydrolases from bacterial proteomes. Using this strategy, the authors identified five peptidoglycan hydrolases from Acinetobacter baumannii, which they tested on various Gram-positive and Gram-negative pathogens for antimicrobial activity. The revised manuscript addressed most of the prior concerns, and the data presented are solid and will be of interest to microbiologists.

eLife assessment

This study presents useful findings on the efficacy and mechanism of linalool protection against Saprolegnia parasitica oomycetes in the grass carp model. The evidence is incomplete since the claims are partially justified, thus there is a need for more experimental data and more rigorous statistical data analysis . Revisions according to the recommendations will improve the work, making it of interest to scientists within the fields of aquaculture, ichthyology, microbiology, and drug discovery.

eLife assessment

This study presents a useful finding on the interplay of CCL5 and miR-324-5p during ischemic stroke injury. Despite its importance, the evidence supporting the claims of the authors is incomplete. In particular, the lack of methodological information, inappropriate statistical testing, a flawed culture system, and the temporal mismatch in the expression of CCL5 and miR-324-5p following stroke have hindered further evaluation of the claims. The work will be of interest to neuroscientists working on brain injury such as stroke.

eLife assessment

This important study investigates the sensitivity to endogenous cosolvents of three families of intrinsically disordered proteins involved with desiccation. The findings, drawn from well-designed experiments and calculations, suggest a functional synergy between sensitivity to small molecule solutes and convergent desiccation protection strategy. The evidence is found to be convincing, and the authors provide appropriate caveats since the study's conclusions are based on a small number of proteins. This work will be of interest to biochemists and biophysicists interested in the conformation-function relationship of intrinsically disordered proteins.

eLife assessment

This study provides an important insight into the mechanisms of cooperation between Hsp70 and its cochaperones during reactivation of aggregated proteins. Based on convincing evidence, the authors demonstrate that the co-chaperone Hsp110 boosts disaggregation activity by enhancing Hsp70 recruitment to protein aggregates. This work is of broad interest to biochemists and cell biologists working in the protein homeostasis field.

eLife assessment

This fundamental study demonstrates a novel method for imaging glutamate receptors in situ via cryo-ET. The use of cutting-edge methods is well-described and is compelling. This paper is broadly relevant to biophysicists and neuroscientists.

eLife assessment

This revised study presents valuable evidence that a combination of endothelial cells, astrocytes, and neuroblastoma cells of human origin can integrate to form an in vitro brain blood barrier, that recapitulates key aspects of its natural counterpart, especially at short times. Convincingly, the mechanism by which neuroblastoma-secreted GDNF increases Claudin-5 and VE-cadherin is described. To substantiate the role of GDNF in vivo, authors demonstrated that knock-down of this neurotrophic factor, increased the permeability of the brain blood barrier in mice. This in vitro system can be used to study the permeability of the human brain blood barrier to different drugs.

eLife assessment

The present study provides valuable evidence on the neurochemical mechanisms underlying working memory in obesity. The authors' approach considering specific working memory operations (maintenance, updating) and putative dopaminergic genes is solid, though the inclusion of a more direct measure of dopamine signaling would have strengthened the work.

eLife assessment

This valuable manuscript reports alterations in autophagy present in dopaminergic neurons differentiated from iPSCs of patients with WDR45 mutations. The authors identified compounds that improved the defects present in mutant cells by generating isogenic iPSC without the mutation and performing an automated drug screening. The methodological approaches are solid, but the claims still need to be completed; showing the effects of the identified compounds on iron-related alterations is crucial. The effects of these drugs in vivo would be a great addition to the study.

eLife assessment

The aim of this valuable study is to identify novel genes involved in sleep regulation and memory consolidation. It combines transcriptomic approaches following memory induction with measurements of sleep and memory to discover molecular pathways underlying these interlinked behaviors. The authors explore transcriptional changes in specific mushroom body neurons and suggest roles for two genes involved in RNA processing, Polr1F and Regnase-1, in the regulation of sleep and memory. Although this work exploits convincing and validated methodology, the strength of the evidence is incomplete to support the main claim that these two genes establish a definitive link between sleep and memory consolidation.

eLife assessment

Through anchored phylogenomic analyses, this important study offers fresh insights into the evolutionary history of the plant diet and geographic distribution of Belidae weevil beetles. Employing robust methodological approaches, the authors propose that certain belid lineages have maintained a continuous association with Araucaria hosts since the Mesozoic era. Although the biogeographical analysis is somewhat limited by uncertainties in vicariance explanations, this convincing study enhances our understanding of Belidae's evolutionary dynamics and provides new perspectives on ancient community ecology.

eLife assessment

This study investigated the involvement of programmed cell death (PCD) in Arabidopsis thaliana root cap cells and its effect on microbial colonization. The authors have reported the importance of timely corpse clearance in the root cap and a root cap-specific transcription factor in controlling microbial colonization by beneficial fungi. By demonstrating the connection between transcriptional control of PCD and microbial colonization, this study provides fundamental insights into how relationships are established and regulated at the root-microbiome interface. The strength of the evidence presented is convincing, providing a foundation for further research concerning the spatial and temporal dynamics of microbiome recruitment along the root axis.

eLife assessment

This important manuscript presents several structures of the Kv1.2 voltage-gated potassium channel, based on state-of-the-art cryoEM techniques and algorithms. The authors present solid evidence for structures of an inactivating mutant of Kv1.2, DTX-bound Kv1.2 and of Kv1.2 in potassium-free solution (with presumably sodium ions bound within the selectivity filter). These structures advance our knowledge of the molecular basis of the slow inactivation process of potassium channels.

eLife assessment

This manuscript describes valuable findings on the expression pattern of orexin receptors in the midbrain and how manipulating this system influences several behaviors, such as context-induced locomotor activity and exploration. The overall strength of evidence - which includes anatomical, viral manipulation studies, and brain imaging - is solid and broadly supports claims in the paper, however, there are several areas in which the conclusions are only partially supported by the statistical evidence. These results have implications for understanding the neural underpinnings of reward and will be of interest to neuroscientists and cognitive scientists with an interest in the neurobiology of reward.

eLife assessment

This is a useful study depicting the ultrastructural features of layer 1 of the human temporal cortex, the authors assess various synaptic parameters, astrocytic volumetric ratio, and mitochondrial morphology. The data were collected using a solid methodology, however, the analysis of the functional vesicle pools is incomplete, and reliance solely on electron microscopy limits the scope of the work to structural observation. The work will be of interest to neuroscientists and computational researchers investigating cortical and network function.

eLife assessment

This study describes the formation of a penetration ring in the rice blast fungus Magnaporthe oryzae during host cell invasion. The work provides useful insights into how the penetration ring facilitates the transition of penetration pegs into invasive hyphae, which leads to a better understanding of plant-pathogen interactions. However, the evidence supporting the function of this novel infection structure remains incomplete and further work is needed to help clarify the exact role of the penetration ring in the infection process.

eLife assessment

This valuable work discusses the phylogenetic conservation of the hippocampal region and primary sensory cortical regions in mammalian species. The authors propose that species-specific differences in behavior and mnemonic functions may be due to differences in cortico-hippocampal connectivity patterns. However, the manuscript, in its present form, is speculative, and the strength of evidence for this proposition is incomplete.

eLife assessment

This research investigates the precision of numerosity perception in two different tasks and concludes that human performance aligns with an efficient coding model optimized for current environmental statistics and task goals. The findings may have important implications for our understanding of numerosity perception as well as the ongoing debate on different efficient coding models. However, the evidence presented in the paper to support the conclusion is still incomplete and could be strengthened by further modeling analysis or experimental data that can address potential confounds.

eLife assessment

This important study reports that slow fluctuations of serotonin release during wakefulness and non-REM sleep correspond to periods of either increased arousal or enhanced offline information processing. The evidence supporting the claim is convincing, and the methodology used in the study will benefit many in the field. The work will be of interest to neuroscientists working on sleep, memory, and neuromodulation.

eLife assessment

This valuable study aims to understand the function of ProSAP-interacting protein 1 (Prosapip1) in the brain. Using a conditional Prosapip1 KO mouse (floxed prosapip1 crossed with Syn1-Cre line), the authors performed analysis including protein biochemistry, synaptic physiology, and behavioral learning. Solid evidence from this study supports a role of Prosapip 1 in synaptic protein composition, synaptic NMDA responses, LTP, and spatial memory. Addressing some of the technical and methodological weaknesses may further improve the significance of the study.

eLife assessment

This study presents a fundamental finding to the field interested in recurrent processing and its neuromodulatory underpinnings, finding unexpectedly that memantine (blocking NMDA-receptors) enhances the decoding of features thought to rely on NMDA-receptors. The evidence is solid and would be improved by further persuading the readership of the likely functional underpinnings of this direction of result and why there was no behavioural effect. These findings will be of interest to a wide community of researchers studying consciousness, sensory processing, attention, and neurotransmitters.

eLife assessment

This study explores the role of protein synthesis in spinal cord neurons in the regulation of chronic pain. Using innovative techniques, this valuable study outlines cell-type specific gene changes that occur in the spinal cord in the early and late phases of nerve injury. The presented evidence and methods used are, however, incomplete: there are several major technical and analysis issues that need to be addressed, and in addition, deeper gene expression analysis and additional controls would have strengthened the conclusions. This work will be of broad interest to biologists studying pathological plasticity in circuits.

Security depends on cryptographically verifiable primitives. Cryptography is native to binary, which makes text-based human readable Protocols (like JSON) awkward.

eLife assessment

This important study reports the discovery of a novel nucleotide ubiquitylation activity by the DTX3L E3 ligase. Solid evidence is presented for ubiquitin attachment to single-stranded oligonucleotides. This very interesting biochemical finding can be used as a starting point for studies to establish relevance in a physiological setting.

eLife assessment

This important study investigates the molecular mechanisms underpinning how the tumor necrosis factor alpha-induced protein, TIPE, regulates aerobic glycolysis to promote tumor growth in melanoma. Solid data using multiple independent approaches provide new insights into the molecular mechanisms underpinning aerobic glycolysis, also known as the Warburg Effect, in melanoma cells. However, further investigation of a potential oncogenic effect of TIPE in melanoma patients is warranted and more advanced metabolomic and bioenergetic assays could be employed. The work will be of interest to biomedical researchers working in cancer and metabolism.

eLife assessment

In this study, Ferling and colleagues provide convincing evidence demonstrating that myeloid cells exert distinct, cargo-dependent responses during and after phagocytosis. These important findings establish previously unrecognized insights into the function(s) of myeloid cells in immunosurveillance and are thus likely to be broadly impactful across the spectrum of biomedical disciplines including immunology and cell biology. Notwithstanding these clear strengths of the article, some minor issues were noted pertinent to the relative opaqueness of the mechanisms underpinning context-specific RhoA activation.

eLife assessment

This important study reports the use of a surveillance approach in identifying emerging diseases, monitoring disease trends, and informing evidence-based interventions in the control and prevention of livestock abortions, as it relates to their public health implications. The data support the convincing finding that abortion incidence is higher during the dry season, and occurs more in cross-bred and exotic livestock breeds. Aetiological and epidemiological data can be generated through established protocols for sample collection and laboratory diagnosis. These findings are of potential interest to the fields of veterinary medicine, public health, and epidemiology.

eLife assessment

This work provides important insight into the mechanisms of hrp2 and particularly hrp3 deletion generation. The generation of additional long-read data alongside a new analysis of 19,000 public short-read sequenced genomes makes this the most detailed investigation currently available on this topic, which has high public health importance and also basic biological interest. The revised version of the manuscript provides convincing evidence for the proposed mechanisms.

eLife assessment

This important work by Malita et al. describes a mechanism by which an intestinal infection causes an increase in daytime sleep through signaling from the gut to the blood-brain barrier. Their findings suggest that cytokines upd3 and upd2 produced by the intestine following infection act on the glia of the blood-brain barrier to regulate sleep by modulating Allatostatin A signaling. The evidence supporting the claims of the authors is solid. Further verification of certain critical tools, and addressing a few discrepancies from data previously published, would improve this work.

eLife assessment

This study presents a valuable dataset regarding chromatin remodeling by the BAF complex in the context of meiotic sex chromosome inactivation. Solid data generally support the conclusions, although the partial deletion of the BAF complex in the germline could be considered limiting. This work will be of interest to researchers working on chromatin and reproductive biology.

eLife assessment

This useful study describes expression profiling by scRNA-seq of thousands of cells of recombinant yeast genotypes from a system that models natural genetic variation. The rigorous new method presented here shows promise for improving the efficiency of genotype-to-phenotype mapping in yeast, providing convincing evidence for its efficacy. This revised manuscript focuses on overcoming technical challenges with this approach and identifies several new biological insights that build upon the field of genotype-to-phenotype mapping, a central question of interest to geneticists and evolutionary biologists.

eLife assessment:

This theoretical study makes a useful contribution to our understanding of a subtype of type 2 diabetes – ketosis-prone diabetes mellitus (KPD) – with a potential impact on our broader understanding of diabetes and glucose regulation. The article presents an ordinary differential equation-based model for KPD that incorporates a number of distinct timescales – fast, slow, as well as intermediate, incorporating a key hypothesis of reversible beta cell deactivation. The presented evidence is solid and shows that observed clinical disease trajectories may be explained by a simple mathematical model in a particular parameter regime.

eLife assessment

This valuable study advances our understanding of the histopathological features of type 1 diabetes, in particular in regard to the composition and spatial organization of pancreas infiltrating immune cells. The evidence supporting the conclusions is convincingly grounded in an application of both state-of-the-art high-dimensional in situ immunostaining technology as well as a tailored image analysis strategy. The work will be of broad interest to type 1 diabetes researchers as it contributes to a better understanding of the disease's etiopathology.

eLife assessment

In the revised version of this important study, the authors present a convincing pipeline for insect genome regulatory annotation across 33 insect genomes spanning 5 orders. Despite technical limitations in the field owing to the lack of comprehensive knowledge of enhancer content in any system, the authors employ several independent downstream analyses to support the validity of their enhancer predictions for a subset of these genomes. Taken together, the revised results suggest that this prediction pipeline may have uses in identifying functional enhancers across large phylogenetic distances. Reviewers note caveats that an experimental validation is not yet available in the field to validate a large class of newly identified enhancers across such evolutionary distances, and other pipelines might be of use to compare. This work will be of interest to the computational genomics, evolutionary biology, and gene regulation fields.

eLife assessment

This manuscript compiles existing algorithms into an open-source software package that enables real-time (and offline) motor unit decomposition from muscle activity collected via grids of surface electrodes and indwelling electrode arrays. The package is valuable given that many motor neuroscience labs are using such algorithms and that there exists a host of potential applications for such data. Validation of the software package is compelling, suggesting that it can be successfully applied across a range of muscles and tasks.

eLife assessment

This study describes a useful antibody-free method to map both G-quadruplexes and R-loops in vertebrate cells independently of the BG4 and S9.6 antibodies. It also reveals that the helicase Dhx9 can affect the self-renewal and differentiation capacities of mESCs, perhaps by regulating co-localized G4s and R-loops. The datasets provided might constitute a good starting point for future functional studies, and although the strength of the evidence that DHX9 interferes with the ability of mESCs to differentiate by regulating directly the stability of either G4s or R-loops has been improved compared to a previous version, it is still incomplete.

eLife assessment

In this useful study, a solid machine learning approach based on a broad set of systems to predict the R2 relaxation rates of residues in intrinsically disordered proteins (IDPs) is described. The ability to predict the patterns of R2 will be helpful to guide experimental studies of IDPs. A potential weakness is that the predicted R2 values may include both fast and slow motions, thus the predictions provide only limited new physical insights into the nature of the underlying protein dynamics, such as the most relevant timescale.

eLife assessment

The manuscript proposes an alternative method by SDS-PAGE calibration of Halo-Myo10 signals to quantify myosin molecules in filopodia and discusses different scenarios regarding myosin 10 working models to explain intracellular diffusion and targeting to filopodia. Overall, the paper is elegantly written and the methodology is valuable in its descriptive potential as these are key numbers to know to ultimately decipher the cellular mechanism of Myo10 action as well as understand the molecular composition of a Myo10-generated filopodium. The evidence for the conclusions is compelling, but there are limitations to this study which should be kept in mind when applying this method to other systems.

eLife assessment

This manuscript reveals an important mechanism of KCNQ1/IKs channel gating and PUFA modulation of this mechanism. This mechanism is supported by convincing single channel recordings, macroscopic current recordings and mutational analyses. These findings are of importance to the ion channel field and possibly future therapeutic applications.

eLife assessment

This study presents a valuable finding on the role of cholesterol-binding site on GLP-1 receptors and functionally characterizes the impact of this mutation on receptor behavior in the membrane and downstream signaling. The computational and experimental approaches used in the study to arrive at the conclusions are solid. The clinical ramifications are unclear at this point, but the study is a helpful addition to the scientific community working on receptor biology and drug development.

eLife assessment

Kewenig et al. present a timely and valuable study that extends prior research investigating the neural basis of abstract and concrete concepts by examining how these concepts are processed in a naturalistic stimulus: during movie watching. The authors provide convincing evidence that the varying strength of the relationship between a word and a particular visual scene is associated with a change in the similarity between the brain regions active for concrete and abstract words. This work makes a contribution that will be of general interest within any field that faces the inherent challenge of quantifying context in a multimodal stimulus.

eLife assessment

This important work, leveraging state-of-the-art whole-night sleep EEG-fMRI methods, advances our understanding of the brain states underlying sleep and wakefulness. Despite a small sample size, the authors present convincing evidence for substates within N2 and REM sleep stages, with reliable transition structure, supporting the perspective that there are more than the five canonical sleep/wake states.

eLife assessment

The methods and findings of the current work are important and well-grounded. The strength of the evidence presented is convincing and backed up by rigorous methodology. The work, when elaborated on how to access the app, will have far-reaching implications for current clinical practice.

eLife assessment

This is an important paper that reports in vivo physiological abnormalities in the hippocampus of a rat model of traumatic brain injury (TBI). In this study, authors focused on changes in theta-gamma phase coupling and action potential entrainment to theta, phenomena hypothesized to be critical for cognition. While the authors provide solid evidence of deficits in both features post-TBI, the study would have been stronger with a more hypothesis-driven approach and consideration of alterations of the animal's behavioral state or sensorimotor deficits beyond memory processes.

eLife assessment

In this valuable study, the authors use deep learning models to provide solid evidence that epithelial wounding triggers bursts of cell division at a characteristic distance away from the wound. The documentation provided by the authors should allow other scientists to readily apply these methods, which are particularly appropriate where unsupervised machine-learning algorithms have difficulties.

eLife assessment

This valuable contribution follows past descriptions of ciliation defects, potentially linked to cholinergic neuronal dysfunction, associated with mutated G2019S Lrrk2 expression. The strength of evidence is considered solid and broadly supportive of the claims concerning well-characterized cilia changes in cholinergic neurons over time in the model; however, additional work may be required to define the specificity of the pRab12 antibody in the IHC technique, dependence on LRRK2, and clarification of the cilia phenotype in sporadic PD brains that exists (for the moment) only in a non-peer-reviewed pre-print, despite the prominence of these (preliminary) results highlighted in the abstract and text of the current manuscript. It is hoped that the authors will begin to address the feedback provided by the expert reviewers to help provide a more mechanistic basis for the audience interested in cholinergic defects associated with Parkinson's disease.

eLife assessment

In this manuscript, Griesius et al analyze the dendritic integration properties of NDNF and OLM interneurons, and the current dataset suggests that even though both cell types display supralinear NMDA receptor-dependent synaptic integration, this may be associated with dendritic calcium transients only in NDNF interneurons. These findings are important because they could shed light on the functional diversity of different classes of interneurons in the mouse neocortex and hippocampus, which in turn can have major implications for understanding information flow in complex neural circuits. They are considered as being currently incomplete, however, due to: (i) the large variability and small sample size of multiple datasets, which prevents a finer evaluation of cellular and molecular mechanisms accounting for the difference in the integrative properties of different interneuron types; (ii) lack of control experiments to rule out that the effect of the NMDA antagonist AP5 on synaptic integration is not confounded by potential phototoxicity damage; (iii) lack of a precise control of the uncaging location.

eLife assessment

This study used electrophysiology and imaging to show that the majority of excitatory cells in the dentate gyrus of adult mice have very slow oscillations during non-rapid eye movement (NREM) sleep. The oscillations were influenced by serotonin when it was released during NREM sleep. Moreover, the serotonin receptor type 1a mediated the effect, and reducing these receptors impaired a type of memory. The significance of the study is important and the strength of the evidence is solid, but revisions to the figures and making conclusions more consistent with the data could improve the significance and strength of evidence.

eLife assessment

This important study by Wong et al. addresses a longstanding question in the field of associative learning regarding how a motivationally relevant event can be inferred from prior learning based on neutral stimulus-stimulus associations. The research provides convincing behavioral and neurophysiological evidence to address this important question. The manuscript will be interesting for researchers in behavioral and cognitive neuroscience.

eLife assessment

This important work combines theory and experiment to assess how humans make decisions about sequences of pairs of correlated observations. The normative theory for evidence integration in correlated environments will be informative for future investigations. However, the developed theory and data analysis seem currently incomplete: it remains to be seen if the derived decision strategy is indeed normative, or only an approximation thereof, and behavioral modelling would benefit from the assessment of alternative models.

Chapter 1 Summary

eLife assessment

This important work identifies a non-autophagic role for ATG5 in lysosomal repair and the trafficking of the glucose transporter GLUT1 to the cell surface, mediated through the retromer complex. The evidence supporting the conclusions is solid.

eLife assessment

Supported by convincing data, this valuable study demonstrates that the Chitinase 3-like protein 1 (Chi3l1) interacts with gut microbiota and protects animals from intestinal injury in laboratory colitis model. The revised manuscript sufficiently addressed the reviewers' comments. The work will be of interest to scientists studying crosstalk between gut microbiota and inflammatory diseases.

eLife assessment

This study provides useful evidence substantiating a role for long noncoding RNAs in liver metabolism and organismal physiology. Using murine knockout and knock-in models, the authors invoke a previously unidentified role for the lncRNA Snhg3 in fatty liver. The revised manuscript has improved and most studies are backed by solid evidence but the study was found to be incomplete and will require future studies to substantiate some of the claims.

eLife assessment

The manuscript by Carbo et al. reports a novel role for the MltG homolog AgmT in gliding motility in M. xanthus. The authors provide convincing data to demonstrate that AgmT is a cell wall lytic enzyme (likely a lytic transglycosylase), its lytic activity is required for gliding motility, and that its activity is required for proper binding of a component of the motility apparatus to the cell wall. The findings are valuable as they contribute to our understanding of the molecular mechanisms underlying the interaction between gliding motility and the bacterial cell wall.

eLife assessment

This useful study describes a single set of label-chase mass spectrometry experiments to confirm the molecular function of YafK as a peptidoglycan hydrolase, and to describe the timing of its attachment to the peptidoglycan. Confirmation of the molecular function of YafK is helpful for further studies to examine the function and regulation of the outer membrane-peptidoglycan link in bacteria. The evidence supporting the molecular function of YafK and that lpp molecules are shuffled on and off the peptidoglycan is solid, however, some of the other data still remain incomplete in the revised version. The work will be of interest to researchers studying lipoproteins in gram negative bacteria.

eLife assessment

This study presents an important finding on the spontaneous emergence of structured activity in artificial neural networks endowed with specific connectivity profiles. The evidence supporting the claims of the authors is convincing, providing direct comparison between the properties of the model and neural data although investigating more naturalistic inputs to the network would have strengthened the main claims. The work will be of interest to systems and computational neuroscientists studying the hippocampus and memory processes.

eLife assessment

This study presents a fundamental finding on how levels of m6A levels are controlled, invoking a consolidated model where degradation of modified RNAs in the cytoplasm plays a primary role in shaping m6A patterns and dynamics, rather than needing active regulation by m6A erasers and other related processes. The evidence is compelling and uses transcriptome-wide data and mechanistic modeling. However, it is possible that m6A-erasers will have roles in specific developmental contexts or conditions, so this model may not apply universally.

eLife assessment

Utilizing transgenic lineage tracing techniques and tissue clearing-based advanced imaging and three-dimensional slices reconstruction, the authors comprehensively mapped the distribution atlas of NFATc1+ and PDGFR-α+ cells in dental and periodontal mesenchyme and tracked their in vivo fate trajectories. This important work extends our understanding of NFATc1+ and PDGFR-α+ cells in dental and periodontal mesenchyme homeostasis, and should provide impact on clinical application and investigation. The strength of this work is compelling in employing CRISPR/Cas9-mediated gene editing to generate two dual recombination systems, and mapped gNFATc1+ and PDGFR-α+cells residing in dental and periodontal mesenchyme, their capacity for progeny cell generation, and their inclusive, exclusive and hierarchical relations in homeostasis, generating a spatiotemporal atlas of these skeletal stem cell population.

eLife assessment

In this fundamental study, the authors describe a new data processing pipeline that can be used to discover causal interactions from time-lapse imaging data. The utility of this pipeline was convincingly illustrated using tumor-on-chip ecosystem data. The newly developed pipeline could be used to better understand cell-cell interactions and could also be applied to perform temporal causal discovery in other areas of science, meaning this work could potentially have a wide range of applications.

eLife assessment

This valuable study characterizes the variability in spacing and direction of entorhinal grid cells and shows how this variability can be used to disambiguate locations within an environment. These claims are supported by solid evidence, yet some aspects of the methodology should be clarified. This study will be of interest to neuroscientists working on spatial navigation and, more generally, on neural coding.

eLife assessment

In this useful study, the authors show that N-acetylation of synuclein increases clustering of synaptic vesicles in vitro and that this effect is mediated by enhanced interaction with lysophosphatidylcholine. While the evidence for enhanced clustering is largely solid, the biological significance remains unclear.

eLife assessment

The manuscript introduces an important and innovative non-AI computational method for segmenting noisy grayscale images, with a particular focus on identifying immunostained potassium ion channel clusters. This method significantly enhances accuracy over basic threshold-based techniques while remaining user-friendly and accessible, even for researchers with limited computational backgrounds. The evidence supporting the method's efficacy is convincing. Its practical application and ease of use make it a tool that will benefit a wide range of laboratories.

eLife assessment

This useful experiment seeks to better understand how memory interacts with incoming visual information to effectively guide human behavior. Using several methods, the authors identify two distinct pathways relating visual processing to the default mode network: one that emphasizes semantic cognition, and the other, spatial cognition. The evidence presented is solid and will be of interest to cognitive and systems neuroscientists.

eLife assessment

This important study shows a significant role for Mushashi-2 (Msi2) in lung adenocarcinoma. The authors provided solid data that support the requirement for Msi2 in tumor growth and progression, although the study would have been strengthened by including more patient samples and additional evidence regarding Msi2+ cells being more responsive to transformation. These findings are of interest to both the lung cancer and the RNA binding protein fields.

eLife assessment

This fundamental study provides compelling evidence for dysgranular insular involvement in top-down and bottom-up interoceptive processing by building on previous evidence using state-of-the-art methods. Its translational application in ADE patients corroborates the assumption that the mid-insula may indeed be a locus of 'interoceptive disruption' in psychiatric disorders, which underscores the study's high relevance for both body-brain as well as clinical research.

eLife assessment

This is a potentially valuable contribution, reporting a deletion analysis of the MSL1 gene to assess how different parts of the protein product interact with the MSL2 protein and roX RNA to affect the association of the MSL complex with the male X chromosome of Drosophila. However, the framework that the MSL complex mediates dosage compensation is outdated and has flaws, and the evidence is currently considered inadequate to support the claims. Because there are many ways to alter viability, sex-specific viability is insufficient to make claims regarding dosage compensation.

eLife assessment

This important study combines genetic analysis, biochemistry, and structural modeling to reveal new insights into how changes in protein-protein structure activate signal transduction as part of the bacterial general stress response. The data, collected using validated and standard methods, and the interpretations are solid, although additional experimental structural evidence would strengthen the proposed model and its potential application to other systems. This manuscript, which provides multiple avenues for follow-up studies, will be of broad interest to microbiologists, structural biologists, and cell biologists.

eLife assessment

This study provides potentially highly valuable new insight into the role of Fgf signalling in SUFU mutation-linked cerebellar tumors and indicates novel therapeutic interventions via inhibition of Fgf signalling. The evidence supporting the major claims, however, is at this point currently incomplete. A more robust analysis of gene expression patterns and deeper mechanistic insight would significantly enhance this study, which could have wide-ranging implications for the treatment of specific cerebellar tumors.

eLife assessment

This manuscript presents a valuable new quantitative crosslinking mass spectrometry approach using novel isobaric crosslinkers. The data are solid and the method has potential for a broad application in structural biology if more isobaric crosslinking channels are available and the quantitative information of the approach is exploited in more depth.

eLife assessment

The authors present a valuable study exploring the interaction between JNK signaling and high sucrose feeding. The strength of evidence supporting these observations is solid, including multi-tissue transcriptomic and metabolic analyses, followed by network modeling approaches to define the organs and pathways involved. Reviewers provided several suggestions to improve the manuscript including clarifications of model and analyses, as well as explanations for within-group variations and confirming RNA-seq results at the level of metabolite processes highlighted.

eLife assessment

Recent studies have demonstrated that depletion of nuclear TDP-43 leads to loss of its nuclear function resulting in changes in gene expression and splicing of target mRNAs. This study developed a sensitive and robust sensor for TDP-43 activity that should impact the field's ability to monitor whether TDP-43 is functional or not. Though limited to cell culture, the evidence presented is convincing and is the first demonstration that a GFP on/off system can be used to assess TDP-43 mutants as well as loss of soluble TDP-43. The findings are valuable and may represent a novel tool to investigate TDP-43-associated disease mechanisms.

eLife assessment

This work describes how the toxin-antitoxin (TA) system, which uses the cyclic di-GMP as an antitoxin, controls both the persistence of antibiotics linked to biofilms and the integrity of the bacterial genome. The authors present solid evidence linking cyclic di-GMP and the toxin HipH. The work is valuable because it establishes the relationship between bacterial persistence and biofilm resilience, which lays a strong basis for future research on the formation of bacterial biofilms and antibiotic resistance.

eLife assessment

This important study evaluates the outcomes of a single-institution pilot program designed to provide graduate students and postdoctoral fellows with internship opportunities in areas representing diverse career paths in the life sciences. The data convincingly show the benefit of internships to students and postdocs, their research advisors, and potential employers, without adverse impacts on scientific productivity. This work will be of interest to multiple stakeholders in graduate and postgraduate life sciences education and should stimulate further research into how such programs can best be broadly implemented.

eLife assessment

Understanding how genomic regulatory elements control spatiotemporal gene expression is essential for explaining cell type diversification, function, and the impact of genetic variation on disease. This important study provides solid evidence that enhancers generally combine additively to influence gene expression. Moreover, promoters, particularly weaker ones, can exhibit supra-additivity when integrating enhancer effects. These findings highlight the context-dependent nature of enhancer-promoter interactions in gene regulation, and contribute to ongoing discussions about the selectivity and combination of regulatory elements.

eLife assessment

This is a valuable contribution to our understanding of how different cell stressors (ethanol or heat-shock) elicit unique responses at the genomic and topographical level under the regulation of yeast transcription factor Hsf1, providing solid evidence documenting the temporal coupling (or lack thereof) between Hsf1 aggregation and long-range communication among co-regulated heat-shock loci versus chromatin remodeling and transcriptional activation. A particular strength is the combination of genomic and imaging-based experimental approaches applied to genetically engineered in vivo systems.

eLife assessment

This manuscript investigates how chloroplasts are broken down during light-limiting conditions as plants reorganize their energy-producing organelles during carbon limitation. The authors provide compelling live-cell imaging data of plastids and solid quantification of events, documenting that buds form on the surface of chloroplasts and pinch away, then associate with the vacuole via a mechanism that depends on autophagy machinery, but not plastid division machinery. This manuscript provides valuable groundwork for other scientists studying the regulation and breakdown of energy-producing organelles, including chloroplasts and mitochondria.

eLife assessment

This study provides valuable insights into the role of actin dynamics in regulating the transition of fusion models during homotypic fusion between late endosomes. The evidence supporting the authors' claims is convincing. However, while the observations are significant, the study could benefit from further exploration of the mechanistic details and physiological relevance.

eLife assessment

This is an important behavioral, pharmacological intervention study of the effects of the catecholamine reuptake inhibitor methylphenidate (MPH) on value-based decision-making using a combination of aversive and appetitive Pavlovian to Instrumental Transfer (PIT) in a human cohort (n=100). The design used drug dosing after learning, allowing the convincing interpretation of catecholamines being involved in the decision process, an effect dependent on baseline working memory capacity. The results also challenge the view that catecholamines operate by modulating behavioural invigoration alone.

eLife assessment

This important study offers a powerful empirical test of a highly influential hypothesis in population genetics. It incorporates a large number of animal genomes spanning a broad phylogenetic spectrum and treats them in a rigorous unified pipeline, providing the convincing negative result that effective population size scales neither with the content of transposable elements nor with overall genome size. These observations demonstrate that there is still no simple, global hypothesis that can explain the observed variation in transposable element content and genome size in animals.

eLife assessment

This valuable study investigates the immune system's role in pre-eclampsia. The authors map the immune cell landscape of the human placenta and find an increase in macrophages and Th17 cells in patients with pre-eclampsia. Following mouse studies, the authors suggest that the IGF1-IGF1R pathway might play a role in how macrophages influence T cells, potentially driving the pathology of pre-eclampsia. There is solid evidence in this study that will be of interest to immunologists and developmental biologists, however, some of the conclusions require additional detail and/or more appropriate statistical tests.

eLife assessment

This important work advances our understanding of how mechanical forces transmitted by blood flow contribute to cardiac development by identifying id2b as a flow-responsive factor that is required for valve development and calcium-mediated cardiac contractility and its downstream mechanism of action. However, the evidence supporting the conclusions is incomplete and would benefit from more rigorous approaches. With additional support of the main conclusions, the work will be of interest to those working on developmental biology, heart development, and congenital heart disease.

eLife assessment

This valuable work analyzes how specialized cells in the auditory cells, known as the octopus cells, can detect coincidences in their inputs at the submillisecond time scale. While previous work indicated that these cells receive no inhibitory inputs, the present study unambiguously demonstrates that these cells receive inhibitory glycinergic inputs. The physiologic impact of these inputs needs to be studied further. It remains incomplete at present but could be improved by addressing caveats related to similar sizes of excitatory postsynaptic potentials and spikes in the octopus neurons.

eLife assessment

This study presents a valuable finding on the role of dopamine receptor D2R in dopaminergic neurons DAN-c1 and mushroom body neurons (Y201-GAL4 pattern) on aversive and appetitive conditioning. The evidence supporting the claims of the authors is solid and promotes the investigation using fly larvae, which have interesting advantages in the time required for obtaining experimental animals and the use of optogenetics. The work will be of interest to researchers studying neuronal control of behaviour and learning and memory in general.

eLife assessment

This important study investigates the relationship between transcription factor condensate formation, transcription, and 3D gene clustering of the MET regulon in the model organism S. cerevisiae. The authors provide solid experimental evidence that transcription factor condensates enhance transcription of MET-regulated genes, but evidence for the role of Met4 IDRs and Met4-containing condensates in mediating target gene clustering in the MET regulon is not as strong. This paper will be of interest to molecular biologists working on chromatin and transcription, although its impact would be strengthened by further investigation.