eLife Assessment

This set of experiments provides a valuable finding regarding the need for prior inhibitory training to recruit the infralimbic cortex in extinction learning. The multiple clever behavioral designs supply converging lines of evidence in a compelling manner, but several issues, such as the group sizes and appropriate analysis of data, render the overall strength of support incomplete. With these issues resolved, this manuscript will be of interest to behavioral neuroscientists, especially those interested in learning & memory and/or cortical function.

eLife Assessment

This important study presents convincing findings on creating an exhaustive library of new enhancer-AAVs targeting astrocytes and oligodendrocytes with high potential for both basic and translational work, which will be of value to a large and growing community. However, the outdated description of glial biology in the Introduction, the overstated claims of utility in the Conclusion, and the loose stringency in the criteria used to assemble the library diminish the strengths of the claims. The work will be of interest to neuroscientists working on glial cell biology.

eLife Assessment

This paper presents a collection of analyses relating structure and function in the whole-brain Drosophila EM connectome and whole-brain calcium imaging data. The linkage of detailed anatomical structure with population activity is of broad interest in circuit neuroscience in light of increasingly detailed brain maps, but the analysis methods used made the evidence incomplete. The conclusions are useful for specific network observations, but a more thorough analysis of the anatomical and functional data is needed to support the overall claims.

eLife Assessment

This study by Lapao et al. uncovers a novel role for the Rab27A effector SYTL5 in regulating mitochondrial function and mitophagy under hypoxic conditions. Using a range of imaging and functional assays, the authors demonstrate that SYTL5 localizes to mitochondria in a Rab27A-dependent manner and impacts mitochondrial respiration and metabolic reprogramming. While the findings are solid and valuable in the area of cancer biology, further mechanistic clarity and improved imaging would strengthen the conclusions.

eLife Assessment

This work provides one of the first important attempts to look at Drosophila immune responses against bacterial, viral, and fungal pathogens in a way that combines the roles of four major arms in immunity (Imd signaling, Toll signaling, phagocytosis, and melanization) rather than studying them separately. The findings are compelling and the tools provided can be used as they are, or built upon, in various contexts.

eLife Assessment

This potentially valuable study presents claims of evidence for coordinated membrane potential oscillations in E. coli biofilms that can be linked to a putative K+ channel and that may serve to enhance photo-protection. The finding of waves of membrane potential would be of interest to a wide audience from molecular biology to microbiology and physical biology. Unfortunately, a major issue is that it is unclear whether the dye used can act as a Nernstian membrane potential dye in E. coli. The arguments of the authors, who largely ignore previously published contradictory evidence, are not adequate in that they do not engage with the fact that the dye behaves in their hands differently than in the hands of others. In addition, the lack of proper validation of the experimental method including key control experiments leaves the evidence incomplete.

eLife Assessment

This is a fundamental study that provides a detailed single-cell transcriptomic and epigenomic map of the mouse trabecular meshwor, identifying three distinct trabecular meshwor subtypes with specific functional roles. It links the glaucoma-associated transcription factor LMX1B to mitochondrial regulation in TM3 cells and demonstrates that nicotinamide treatment prevents IOP elevation in Lmx1bV265D/+ mutant mice, highlighting a potential metabolic therapeutic strategy for glaucoma. This convincing work would be further supported by data that link the transcriptional data with mitochondrial functional assays.

eLife Assessment

This is a valuable methodological contribution towards accurate characterization of viral genetic diversity using long-read sequencing and unique molecular identifiers (UMIs). However, the methods are currently incomplete and the sensitivity is not rigorously demonstrated. Addressing these gaps would strengthen the manuscript and make it a key addition to the field.

eLife Assessment

This study provides important insights into the role of polyUbiquitination in neurodegenerative diseases, elucidating how pUb promotes neurodegeneration by affecting proteasomal function. The findings not only offer a new perspective on the pathophysiology of neurodegenerative diseases but also provide potential targets for developing new therapeutic strategies. The experiments in the revised submission provide solid evidence to support the conclusions.

eLife Assessment

This manuscript presents an in-depth analysis of gene expression across multiple brown algal species with differing life histories, providing convincing evidence for the conservation of life cycle-specific gene expression. While largely descriptive, the study is an important step forward in understanding the core cellular processes that differ between life cycle phases, and its findings will be of broad interest to developmental and evolutionary biologists.

eLife Assessment

In this preregistered study, Kunkel and colleagues set out to compare the magnitude and duration of placebo versus nocebo effects in healthy volunteers, and also to examine the different factors contributing to these effects. The authors follow a rigorous methodology in a within-subjects design, taking into consideration standard conventions for manipulation of expectations, and using an appropriate sham condition. They present compelling evidence of long-lasting placebo and nocebo effects, with nocebo responses demonstrating consistently greater strength. These valuable results have the potential for a great impact in the field of experimental and clinical pain.

eLife Assessment

This work describes a highly complex automated algorithm for analyzing vascular imaging data from two-photon microscopy. This tool has the potential to be extremely valuable to the field and to fill gaps in knowledge of hemodynamic activity across a regional network. The solid biological application provides a demonstration of their pipeline's capabilities and suggests intriguing hypotheses around prolonged vascular tone changes, but will need to be followed up by further experiments to be conclusively demonstrated.

eLife Assessment

This paper provides important insight into how early life experience shapes adult behavior in fruit bats. The authors raised juvenile bats either in an impoverished or enriched environment and studied their foraging behaviors. The evidence is convincing that bats raised in enriched environments are more active, bold, and exploratory. The work will be of interest to ethologists and developmental psychologists.

eLife Assessment

This revised manuscript presents an important characterization of mouse auditory cortex receptive field organization, utilizing two-photon imaging of specific subpopulations. They demonstrate a degradation of tonotopic organization from the input to the output neurons. The strength of the evidence is convincing.

eLife Assessment

This study presents a valuable assessment of and solid evidence for increased similarity in visual appearance combined with increased chemical differences between two butterfly species in sympatry compared with differences between three populations of one of the two species in allopatry. The similarity in visual appearance hints to an evolutionary response to shared predators (but alternative explanations are possible). Thus, the difference in chemical signaling likely helps to avoid between-species mating in sympatry.

eLife Assessment

This study presents important information about the role of mu opioid receptors in neurotransmission between the medial habenula and the interpeduncular nucleus. The authors provide convincing evidence that mu opioid receptor activation has differential effects on transmission from substance P neurons and cholinergic neurons, and that blockade of potassium channels can unmask a nicotinic cholinergic synaptic response. This work will be of high interest to those studying this brain region, and potentially to the larger neuroscience community studying motivated behavior.

eLife Assessment

This important study elucidates the role of the exocyst component EXOC6A at distinct stages of ciliogenesis, which advances our understanding of ciliary membrane remodeling and cilium formation. The authors provide solid evidence that EXOC6A interacts with myosin-Va and is dynamically recruited via dynein-, microtubule-, and actin-dependent mechanisms, to support proper formation of the ciliary membrane. The study will be of interest to cell biologists and other researchers interested in vesicular trafficking, organellar membrane dynamics, and ciliogenesis.

eLife Assessment

This valuable study investigates the role of HIF1a signaling in epicardial activation and neonatal heart regeneration in mice. Using a combination of genetic and pharmacological approaches, the authors demonstrate that stabilization of HIF1a enhances epicardial activation and extends the regenerative capacity of the heart beyond the typical neonatal window following myocardial infarction. The main conclusion is well supported by solid data, although some minor concerns regarding experimental interpretation require further clarification to ensure accuracy.

eLife Assessment

This fundamental work substantially advances our understanding of how the glycocalyx of cells provide a non-specific barrier for the interaction of viruses with cell-surface receptors. Using both in vitro experiments and in vivo manipulations they provide compelling evidence for the properties of the glycocalyx to serve as an energy barrier as a main attribute of its mode of action. The work will be of broad interest to virologists and the cell biology community that studies host-pathogen interactions.

eLife Assessment

Endothelial cell-specific loss of TGF-beta signaling in mice leads to CNS vascular defects, specifically impairing retinal development and promoting immune cell infiltration. The data are solid, showing that loss of TGF-beta signaling triggers vascular inflammation and attracts immune cells specific to CNS vasculature. These findings are important, highlighting TGF-beta's role in maintaining vascular-immune homeostasis and its therapeutic potential in neurovascular inflammatory diseases.

eLife Assessment

This cleverly designed and potentially important work supports our understanding regarding how and whether social behaviours promoting egalitarianism can be learned, even when implementing these norms entails a cost for oneself. However, the evidence supporting the major claims is currently incomplete, with the major limitation being whether Ps truly learn egalitarianism from a teacher or instead exhibit reduced guilt across time that is reduced when observing others behaving more selfishly. With a strengthening of the supporting evidence, this work will be of interest to a wide range of fields, including cognitive psychology/neuroscience, neuroeconomics, and social psychology, as well as policy making.

eLife Assessment

This important study provides solid evidence for new insights into the role of Type-1 nNOS interneurons in driving neuronal network activity and controlling vascular network dynamics in awake, head-fixed mice. The authors use an original strategy based on the ablation of Type-1 nNOS interneurons with local injection of saporin conjugated to a substance P analogue into the somatosensory cortex. They show that ablation of type I nNOS neurons has surprisingly little effect on neurovascular coupling, although it alters neural activity and vascular dynamics.

eLife Assessment

This paper addresses a significant question regarding the low overlap between genetic discoveries for human complex diseases and those for gene expression by emphasizing the contribution of cell-type-specific chromatin accessibility QTLs. The analyses supporting the main claims are convincing, and the key conclusions are valuable and of interest to readers in the fields of human genetics and functional genomics.

eLife Assessment

This study reports important findings about the nature of feedback to primary visual cortex (V1) during object recognition. The state-of-the-art functional MRI evidence for the main claims is solid, and the combination of high-resolution fMRI with MEG yields significant insight into neural mechanisms. The findings presented here are relevant to a number of scientific fields such as object recognition, categorisation and predictive coding.

eLife Assessment

This important study identifies a plant-derived metabolite, betulin, as an effective natural insecticide against aphids and uncovers its specific molecular target. The evidence is compelling, combining greenhouse and field efficacy trials with rigorous molecular, genetic, and electrophysiological approaches that converge on a conserved binding site in the aphid GABA receptor. While additional work is needed to fully assess potential off-target effects and ecological safety, the study provides a strong mechanistic foundation. These findings will be of interest to researchers in plant biology, chemical ecology, and sustainable pest management.

eLife Assessment

This study presents a valuable finding about how receptor-ligand binding pathways with multi-site phosphorylation can show non-monotonic responses to increasing ligand affinity and to kinase activity. The authors provide compelling evidence through a simple ordinary differential equation model of such signaling networks with the key new ingredient of ligand-induced receptor degradation. The work will be of interest to physicists and biologists working on signal transduction and biological information processing.

eLife Assessment

This important study characterizes and validates a new activity marker - fast labelling of engram neurons (FLEN) - which is transiently active and driven by cFos, allowing the monitoring of intrinsic and synaptic properties of engram neurons shortly after the learning experience. The results convincingly demonstrate the utility of this novel viral tool for studying early changes in the properties of engram cells. FLEN will provide a beneficial tool for the neuroscience community once it is made available at a plasmid repository.

eLife Assessment

This work derives a valuable general theory unifying theories of efficient information transmission in the brain with population homeostasis. The general theory provides an explanation for firing rate homeostasis at the level of neural clusters with firing rate heterogeneity within clusters. Applying this theory to the primary visual cortex, the authors present solid evidence that accounts for stimulus-specific and neuron-specific adaptation. Reviewers have provided additional suggestions for improving the readability of the manuscript, as well as discussing previous results on adapting coding as well as those aspects of experimental data that are not fully explained by the present theory.

eLife Assessment

This important study investigates nerve-injury-induced allodynia by studying the role of a subpopulation of excitatory dorsal horn CCK+ neurons that express the estrogen receptor GPR30 and potentially modulate nociceptive sensitivity via direct inputs from primary somatosensory cortex. In this revised version, the authors addressed many of the critiques raised through added analyses that convincingly support the notion that spinal GPR30 neurons are indeed an excitatory subpopulation of CCK+ neurons that contribute to neuropathic pain. While evidence of a direct functional corticospinal projection to CCK+/GPR30+neurons is not fully demonstrated, this work will be of broad interest to researchers interested in the neural circuitry of pain.

eLife Assessment

This study presents a valuable finding regarding the role of oxytocin neurons in thermogenesis and behavioral thermoregulation. The use of numerous converging methods, including behavior, fiber photometry, optogenetics, thermal recordings, metabolic analyses, and more, produces a multi-dimensional dataset delivering findings that provide solid support for the conclusions. Conclusions would be strengthened with validation of the approaches, inclusion of a loss of function experiment, and further investigation of the social nature of the behavior. The maternal findings are, at present, somewhat disconnected from the conclusions. The findings are novel and open new doors for understanding the role of the PVT and oxytocin in thermoregulation work; the work will be of strong interest to the thermoregulation, social behavior, and oxytocin signaling communities.

eLife Assessment

This valuable study identifies a brown adipose tissue-specific heat shock factor 1-alcohol dehydrogenase 5 (ADH5) molecular cascade as a regulator of systemic aging, showing that ADH5 deficiency contributes to BAT dysfunction and health decline in aged mice. While there is evidence to support this mechanism, the conclusions remain incomplete, particularly regarding statistical rigor and clarity in data presentation.

eLife Assessment

This modelling study tests several hypotheses describing how seasonality and migration drive the epidemiology of Rift Valley Fever Virus among transhumant cattle in The Gambia. The work is methodologically solid, and findings offer valuable insights into how the movement of cattle in and out of the Gambia River and Sahel ecoregions could lead to source-sink transmission dynamics among cattle subpopulations, sustaining endemic transmission.

eLife Assessment

This important study identifies a putative iron and zinc transporter in the plasma membrane of the obligate intracellular pathogen, Toxoplasma gondii. Using an array of different approaches, the authors convincingly demonstrate that this transporter regulates diverse cellular processes, including parasite metabolism and differentiation. This work will be of broad interest to cell biologists and biochemists studying metal ion transport mechanisms.

eLife Assessment

In this manuscript, the authors report the fundamental finding that a secreted ubiquitin ligase of Shigella, called IpaH1.4, mediates the degradation of a host defense factor, RNF213. The data are convincing and represent a major contribution to our understanding of cell-autonomous immunity and bacterial pathogenesis as they provide new mechanistic insight into how the cytosolic bacterial pathogen Shigella flexneri evades IFN-induced host immunity.

eLife Assessment

This study presents important methodologies for repeated brain ultrasound localization microscopy (ULM) in awake mice and a set of results indicating that wakefulness reduces vascularity and blood flow velocity. The data supporting these findings are solid. This study is relevant for scientists investigating vascular physiology in the brain.

eLife Assessment

This important study investigates why the 13-lined ground squirrel (13LGS) retina is unusually rich in cone photoreceptors, the cells responsible for color and daylight vision. The authors perform deep transcriptomic and epigenetic comparisons between the mouse and the 13-lined ground squirrel (13LGS) to provide convincing evidence that identifies mechanisms that drive rod vs cone-rich retina development. Overall, this key question is investigated using an impressive collection of new data, cross-species analysis, and subsequent in vivo experiments. However, the functional analysis showing the sufficiency and necessity of Zic3 and Mef2C remains incomplete, and further analyses are needed to support the claim that these enhancers are newly evolved in 13LGS.

eLife Assessment

This important study shows that calcium stores in the endoplasmic reticulum of the parasitic protozoan, Toxoplasma gondii play a major role in buffering calcium levels in the cytosol as well as other organelles such as the mitochondrion. Advanced imaging techniques, including use of genetically encoded calcium indicators provide compelling evidence for the role of the SERCA-Ca2+ ATPase pump in regulating organellar calcium levels. However, it remains unclear whether intra-organellar calcium transport occurs via ER-mitochondria membrane contact sites or other mechanisms. This work will be of interest to cell and molecular biologists interested in calcium signalling in divergent eukaryotes.

eLife Assessment

Whole-brain imaging of neuronal activity in freely behaving animals holds great promise for neuroscience, but numerous technical challenges limit its use. In this important study, the authors describe a new set of deep learning-based tools to track and identify the activity of head neurons in freely moving nematodes (C. elegans) and jellyfish (Clytia hemisphaerica). While the tools convincingly enable high tracking speed and accuracy in the settings in which the authors have evaluated them, the claim that these tools should be easily generalizable to a wide variety of datasets is incompletely supported.

eLife Assessment

This study makes a novel and valuable contribution by adapting step selection functions, traditionally used in animal ecology, to explore human movement and environmental risk exposure in urban slums, offering a promising framework for spatial epidemiology, particularly regarding leptospirosis. The integration of GPS telemetry with environmental data and the stratification by gender and serostatus are notable strengths that enhance the study's relevance for public health applications. The strength of evidence is compelling.

eLife Assessment

This valuable study presents a theoretical model of how punctuated mutations influence multistep adaptation, supported by empirical evidence from some TCGA cancer cohorts. This solid model is noteworthy for cancer researchers as it points to the case for possible punctuated evolution rather than gradual genomic change. However, the parametrization and systematic evaluation of the theoretical framework in the context of tumor evolution remain incomplete, and alternative explanations for the empirical observations are still plausible.

eLife Assessment

This important work fills a gap in the characterization of motor architecture and chemical coupling of the male reproductive system, crucial to understanding male reproduction and fertility. The convincing analysis reveals two distinct types of glutamatergic neurons that co-release either serotonin or octopamine. While serotonergic neurons are required for male fertility, octopaminergic neurons are dispensable, indicating a division of labour. This work lays the foundations for future investigations into the conserved key principles by which multi-transmitter systems control coordinated motor outputs.

eLife Assessment

This useful study uses creative scalp EEG decoding methods to attempt to demonstrate that two forms of learned associations in a Stroop task are dissociable, despite sharing similar temporal dynamics. However, the evidence supporting the conclusions is incomplete due to concerns with the experimental design and methodology. This paper would be of interest to researchers studying cognitive control and adaptive behavior, if the concerns raised in the reviews can be addressed satisfactorily.

eLife Assessment

This important study used five metrics to compare the cost-effectiveness of intramural and extramural research funded by the National Institutes of Health in the United States between 2009 and 2019. They found that each type of research had its own set of strengths: extramural research was more cost-effective in terms of publications, whereas intramural research was more cost-effective in terms of influencing clinical work. The evidence supporting these findings is mostly solid, but there are a number of questions about the methods and data - notably about indirect cost recovery and other non-NIH sources of funding - that need to be answered.

eLife Assessment

This paper presents important new findings about the impact of the TAK-003 vaccine against dengue based on a convincing reanalysis of trial data. The results corroborate those of the original trial analyses, but with reduced uncertainty about the estimates of the impact of the vaccine. The findings will be of interest to clinicians, infectious disease epidemiologists, trial statisticians and policymakers seeking to understand the vaccine's efficacy profile and associated uncertainties.

eLife Assessment

This study makes a valuable contribution by separating two timescales of adaptation: rapid, within block reductions in learning rate, and slower, location specific, meta-learned adjustments. Behavioural data and computational modeling converge to support both processes. The evidence is solid with neuroimaging results suggesting that meta-learned learning rates are encoded in the orbitofrontal cortex, while prediction errors are represented in a distributed network including the ventral striatum and are modulated by expected error magnitude, though the specificity of these effects requires further contextualization. The manuscript is timely and clearly written; its main limitation is the weak linkage between neural signals and behavior, leaving uncertainty over whether the reported signals play a mechanistic role in learning.

eLife Assessment

The authors make an important advance in enzyme annotation by fusing biochemical knowledge with language‑model-based learning to predict catalytic residues from sequence alone. Squidly, a new ML method, outperforms existing tools on standard benchmarks and on the CataloDB dataset. The work has solid support, yet clarifications on dataset biases, ablation analyses, and uncertainty filtering would strengthen its efficiency claims.

eLife Assessment

This work demonstrates an objective way to select parameter values for a quadratic integrate-and-fire model so that its bifurcation diagram matches a specific target diagram, generated from the Wang-Buzsaki model. The method is useful for the field and is presented with convincing evidence. The method is currently limited in its ability to be applied to data, but improves our mathematical tools to treat a rarely studied type of bifurcation.

eLife Assessment

In this important study, the authors conducted extensive atomistic and coarse-grained simulations as well as a lattice Monte Carlo analysis to probe the driving force and functional impact of supercomplex formation in the inner mitochondrial membrane. The study highlighted the major contribution from membrane mechanics to the supercomplex formation and revealed interesting differences in structural and dynamical features of the protein components upon complex formation. Upon revision, the analysis is considered solid, although the magnitude of estimated membrane deformation energies seem somewhat surprisingly large. Overall, the study is thorough, creative and the impact on the field of bioenergetics is expected to be significant.

eLife Assessment

This is an important study describing the morphological changes during boundary formation between sensory and non-sensory tissues of the inner ear. The authors provided solid evidence that a transcription factor, Lmx1a and ROCK-dependent actinomyosin are key for border formation in the inner ear. However, future studies will be needed to investigate the direct relationships among boundary formation, Lmx1a and ROCK. This work will be of interest to developmental biologists interested in boundary formation.

eLife Assessment

This valuable study expands the inventory of polyadenylated RNAs cleaved by the double-stranded RNA endonuclease Rnt1 in budding yeast, using solid methodology based on high-throughput sequencing. Previous studies had anecdotally discovered mRNA substrates, and this global characterization is comprehensive with multiple complementary controls. This study sets the stage for deeper investigations into the biological function of Rnt1 and substrate cleavage.

eLife Assessment

This study combines mathematical models and experimental data to analyse the emergence of heterogeneity within clonal NK cell responses during antigen-specific cell expansion. It comprises different experimental data and extensively explores various mathematical models, to study NK cell turnover during acute immune responses and homeostatic turnover within murine cytomegalovirus infection (MCMV). The solid study presents valuable findings and provides insights on heterogeneous NK cell development

eLife Assessment

This study combines genetic, cell biological, and interaction data to propose a model of meiotic double-strand break regulation in C. elegans. Solid evidence supports the main conclusions, while by nature of a screening-type study, more may be needed to solidify speculations in future studies. Yet, comprehensive cataloging of the physical and genetic interactions of factors required for meiotic double-strand break is useful information for the field.

eLife Assessment

The study provides valuable insights into the role of thalamic nuclei in associative threat and extinction learning, supported by a large dataset and multipronged analyses. However, aspects of the evidence remain incomplete, particularly regarding the statistical methods, the claims of plasticity, and the network modeling framework. With this addressed, this manuscript will be of interest to those interested in learning and memory, fear, thalamic circuitry, and related mental heath conditions.

eLife Assessment

In this manuscript, the authors used in vivo long-term tip recordings of the long trichoid sensilla of male hawkmoths to analyze spontaneous spiking activity indicative of the ORNs' endogenous membrane potential oscillations. The authors combine extracellular electrophysiology of the hawkmoth antennae with computational modeling to predict that Orco receptor neuron (ORN) activity is required for circadian, not ultradian, firing patterns. The work provides valuable support for the hypothesis that a posttranslational feedback loop regulates daily and ultradian rhythms in neuronal excitability. Nevertheless, the evidence reported provides only incomplete support for their conclusions, especially with regard to the biological implications of their assumption-heavy models.

eLife Assessment

This study presents valuable new insights into the patterns of organelle inheritance in the protozoan parasite Toxoplasma gondii. An innovative dual-labeling approach used in this study to track maternal-derived and de novo synthesized organelles provides a technical advance with potential to be more broadly applied. Solid evidence is provided that different organelles show distinct inheritance fates during cell replication; however, the data describing the residual body component in this process is incomplete.

eLife Assessment

This is a solid paper on intermittent fasting that will be of interest to readers. The data presented are certainly valuable as a resource. The findings of both shared and tissue-specific signatures, both at the proteomic and transcriptomic levels, align well with what has been established and bring new insight into metabolic adaptation and its consequences in muscle, cortex, and liver.

eLife Assessment

This important study provides evidence for our understanding of HIV transmission dynamics by age and sex in Zambia during the PopART trial; by combining phylogenetic and individual-based mathematical modelling (IBM), it adds depth to the epidemiological literature and may inform more strategic allocation of HIV prevention resources in sub-Saharan Africa. The authors employ two complementary and well-established methodologies (phylogenetics and IBM), and this dual approach is a notable strength. However, the evidence supporting key conclusions is incomplete, with several claims insufficiently substantiated by the data presented. Improvements in data presentation (e.g., quantification of qualitative statements, statistical estimates, and clearer description of results) would substantially strengthen the paper.

eLife Assessment

This is a useful paper regarding the roles of brown adipose tissue and skeletal muscle in thermogenesis in mice, with potential significance for the field. The overall approach is innovative but on balance the evidence for the claim is incomplete, as cast immobilization, while innovative, is likely stressful, may impact muscle and BAT directly, and imposes an energetic cost of motion on the animal that is not accounted for. Further experiments are also needed to directly assess the role of adipose-derived BCAAs in thermogenesis. The authors have done a good job of textually editing their manuscript to clarify the findings and limitations of the study.

eLife Assessment

This useful study provides new insights into the liver stage antigen LSA3, its export to erythrocytes, and its role in liver stage development. While the functional importance of LSA3 is well-demonstrated, the data underlying conclusions about antibody specificity, liver stage localization, and phenotype remain incomplete. A key gain is the use of mosquito and humanized mouse models to access life cycle stages rarely studied in most laboratories.

eLife Assessment

This work, combining behavioural genetics and calcium imaging, provides evidence for a form of learning in Drosophila that derives solely from direct or (optogenetically induced) phantom experience of punishment or reward. Flies that experience foot-shock alone show a subsequent decrease in avoidance to all odorants, together with increased odor-evoked activation of reward-encoding dopaminergic neurons that innervate the mushroom body. Phantom reward, delivered via optogenetic activation of reward-encoding dopaminergic neurons, increases subsequent odour-avoidance. While the findings are valuable to the field, there are aspects of the work that are incomplete, and some of the conclusions and terminology are also not completely justified; three major issues include : (a) the use of the term "priming" to describe this form of learning seems inappropriate and inconsistent with the accepted definition of this term; (b) a key 1998 publication with an initial description of this behavioural phenomenon needs to be cited and presented as context; and (c) the work on reward induced increase in odor-aversion seems relatively preliminary.

eLife Assessment

Liang et al. have conducted a small pilot study investigating the feasibility and tolerability of a regimen of neoadjuvant chemo-immunotherapy for non-small cell lung cancer, with lower cumulative dose of chemotherapy and with the immunotherapy delivered on D8 of each cycle. The clinical data are interesting and novel, and overall the findings of the study are valuable. However, the translational data and analyses are incomplete and do not support key claims in the title.

eLife Assessment

Glioblastoma is among the most aggressive cancers without a cure, and its cells are characterized by high mitochondrial membrane potential. This manuscript provides solid evidence that glioblastoma tumorigenesis is closely linked to mitochondrial stress. The study makes a valuable contribution to the field by advancing our understanding of the metabolic mechanisms driving glioblastoma and highlighting potential therapeutic targets.

eLife Assessment

This manuscript by Kaur et al. identifies differential gene expression observed in distinct cell populations, namely myeloid and lymphoid cells, upon short-term exposure to e-cig aerosols with various flavors. Their findings are useful because they provide a single cell sequencing data resource for assessing which genes and cellular pathways are most affected by e-cig aerosols and their components. However, the evidence is incomplete due to limited analyses and replicates per condition, as well as the lack of in vivo validation.

eLife Assessment

This fundamental study uncovers the unique molecular features of Arabidopsis phloem companion cells that highly express FLOWERING LOCUS T (FT). These FT-expressing cells constitute a distinct subpopulation marked by elevated ATP biosynthesis and co-expression of small mobile proteins such as FLP1 and BFT, highlighting a fine balance between florigen and anti-florigen signals. Motif analyses and transgenic studies further identify NIGT1 transcription factors as direct, nitrogen-inducible repressors of FT, providing a mechanism for delayed flowering under nitrogen-rich conditions. Together, the compelling findings show that florigen-producing companion cells integrate energy metabolism, systemic protein signals, and nutrient-responsive repression to fine-tune the seasonal and nutritional regulation of flowering.

eLife Assessment

This manuscript presents an important finding that D1- and D2-striatal neurons receive distinct cortical inputs, offering key insights into corticostriatal function. For instance, in the context of striatal-dependent learning, this distinction is highly informative for interpreting synaptic physiology data, particularly when inputs to one neuron subtype may change independently of the other. The strength of the evidence is solid, with anatomical and electrophysiological findings aligning well with results from optogenetic and behavioral studies. The study would be of interest to neuroscientists studying basal ganglia circuits in health and disease.

eLife Assessment

This work introduces a new Python package, Avian Vocalization Analysis (AVN) that provides several key analysis pipelines for birdsong research. This tool is likely to prove useful to researchers in neuroscience and beyond, as demonstrated by convincing experiments using a wide range of publicly available birdsong data.

eLife Assessment

This valuable study tests whether prediction error or prediction uncertainty controls how the brain segments continuous experience into events. The paper uses validated models that predict human behavior to analyze multivariate neural pattern changes during naturalistic movie watching. The authors provide solid evidence that there are overlapping but partially distinct brain dynamics for each signal.

eLife Assessment

This manuscript advances the prior finding that antigen recognition in the skill helps establish skin resident memory in CD8 T cells by elucidating the role of TGFBR3 in regulating CD8+ TRM skin persistence upon topical antigen exposure. Key novelty of the your work lies in generation and use of the CD8+ T cell-specific TGFBR3 knockout model, which allows them to demonstrate the role of TGFBR3 in fine tuning the degree of CD8+ T cell skin persistence and that TGFBR3 expression is promoted by CD8+ TRM encountering their cognate antigen upon initial skin entry. This is an important finding and is supported by convincing evidence. There are concerns about the use of FTY720 and the need to establish active TGFbeta limiting conditions to further test this working model.

eLife Assessment

This study addresses an important question and shows how social navigation in homing pigeons can be explained by simple averaging, without requiring any complex cognitive abilities. The evidence, based on a rigorous and systematic comparison of seven models and data on how social routes can be generated from solitary routes, is compelling. The authors should be commended for their willingness to critically re-examine established interpretations.

eLife Assessment

This valuable study examines the cleavage of motor neuron nucleoporins by proteases 2A and 3C of enterovirus D68, a pathogen associated with acute flaccid myelitis. The evidence supporting the effects of EV-D68 proteases on nuclear import and export is solid and confirms previous results on the specific targeting of nucleoporins by proteases from other enteroviruses. However, the claim that cleavage of nucleoporins by EV-D68 2A is neurotoxic, though intriguing, is incomplete, as the evidence is largely indirect.

eLife Assessment

This useful study has demonstrated that MORC2 undergoes phase separation in cells and established multiple interactions responsible for the phase separation. While the characterizations of protein-protein and protein-DNA interactions are solid, there is currently incomplete evidence supporting the claim that MORC2 phase separation contributes to the gene regulatory role of MORC2 in cells. With a stronger link between MORC2 phase separation and cellular function, and further analysis of how disease-linked mutations impact condensation propensity, this study would be of significant interest to biophysicists and molecular biologists working on the role of condensates in gene regulation.

eLife Assessment

This important study demonstrates that ocular organoids can generate both retina and lens through a non-canonical, "inside-out" morphogenetic route. The work is solid, with well-designed experiments combining imaging, molecular analyses, and transcriptomics to establish that lens formation in organoids follows conserved molecular programs despite an alternative morphogenesis. These findings expand our understanding of self-organization and developmental plasticity, and will be of broad interest to researchers working on eye development, organoids, and tissue engineering.

[Editors' note: this paper was reviewed by Review Commons.]

eLife Assessment

This important work examines how microexons contribute to brain activity, structure, and behavior. The authors find that loss of microexon sequences generally has subtle impacts on these metrics in larval zebrafish, with few exceptions. The evidence is convincing, using modern high-throughput phenotyping methodology in zebrafish. Overall, this work will be of interest to neuroscientists and generate further studies of interest to the field.

eLife Assessment

This Review Article explores the intricate relationship between humans and Mycobacterium tuberculosis (Mtb), providing an additional perspective on TB disease. Specifically, this review focuses on the utilization of systems-level approaches to study TB, while highlighting challenges in the frameworks used to identify the relevant immunologic signals that may explain the clinical spectrum of disease. The work could be further enhanced by better defining key terms that anchor the review, such as "unified mechanism" and "immunological route." This review will be of interest to immunologists as well as those interested in evolution and host-pathogen interactions.

eLife Assessment

This study uses all-optical electrophysiology methods to provide a valuable insight into the organization of cortical networks and their ability to balance the activity of groups of neurons with similar functional tuning. The all-optical approach used in this study is impressive and the claim that the effects of optical stimulation correspond to a specific homeostatic mechanism is solid. The work will be of interest to neurobiologists and to developers of optical approaches for interrogating brain function.

eLife Assessment

This study presents a useful method based on flow cytometry to study partitioning noise during cell division. The methods, data and analysis support the claims of the authors is convincing. This work will be of interest to cell biologists and biophysicists working on asymmetric partitioning during cell division.

eLife Assessment

This important study addresses mechanisms of feedback inhibition between planar cell polarity protein complexes during convergent extension movements in Xenopus embryos. The authors propose a conceptually new model, in which non-canonical Wnt ligand stimulates transition of Dishevelled from its complex with Vangl to Frizzled, with essential roles of Prickle and Ror in this process. The main observations supporting molecular interactions are interesting and convincing but do not directly assess convergent extension, and the immunoprecipitations carried out with overexpressed proteins show subtle effects. With the analysis of cell intercalations supporting the main conclusions, this work would be significant and of broad interest to cell and developmental biologists.

eLife Assessment

This work presents a useful investigation of functional and structural brain changes following navigation and verbal memory training. The analyses of whole-brain volumetric changes are convincing and support the study's main conclusion regarding the lack of a volumetric whole-brain plasticity effects. Some analyses are compelling in demonstrating the presence of longitudinal behavioural effects, the presence of functional activation changes, and the lack of hippocampal volume changes.

eLife Assessment

In this manuscript the authors examine correlations between intrinsic electrophysiological properties of HVC neurons projecting to Area X and the temporal structure of the birds' song. The study provides important insights into how the structure of vocalization can relate to intrinsic physiological properties of the neurons that are essential for learning the behavior. The evidence supporting the idea that song temporal structure is related to intrinsic physiology is solid and this research will be of general interest to researchers in the field and neurophysiologists.

eLife Assessment

In this important study, the authors provide compelling evidence that the likelihood of looking behaviour is predicted by the expected information gain, hence constituting an invaluable formal model and explanation of habituation. Such modelling represents a crucial advance in explanation, over-and-above less specified models that can be fitted post hoc to any empirical pattern. The findings would be of interest to researchers studying cognitive development, and perception and learning more broadly.

eLife Assessment

This important work presents an example of how genomic data can be used to improve understanding of an ongoing, long-term bacterial outbreak in a hospital with an application to multi-drug resistant Pseudomonas aeruginosa, and will be of interest to researchers concerned with the spread of drug-resistant bacteria in hospital settings. The convincing genomic analyses highlight the value of routine surveillance of patients and environmental sampling and show how such data can help in dating the origin of the outbreak and in characterising the epidemic lineages. These findings highlight the importance of understanding environmental factors contributing to the transmission of P. aeruginosa for guiding and tailoring infection control efforts.

eLife Assessment

This is an important study demonstrating that cholecystokinin is a key modulator of auditory thalamocortical plasticity during development and in young adult but not aged mice, though cortical application of this neuropeptide in older animals appears to go some way to restoring this age-dependent loss in plasticity. A strength of this work is the use of multiple experimental approaches, which together provide convincing support for the proposed involvement of cholecystokinin. This work is likely to be influential in opening up a new avenue of investigation into the roles of neuropeptides in sensory plasticity.

eLife Assessment

This valuable study explores the role of spatial genome organization in oncogenic transformation, addressing an ambitious and significant topic. The authors have assembled comprehensive datasets from various subtypes of localized and lung-metastatic breast cancer cells, as well as from healthy and cancerous lung cells. They identified switching patterns in the 3D genome organization of lung-metastatic breast cancer cells, revealing a reconfiguration of genome architecture that resembles that of lung cells. This provides solid evidence with significant biomedical implications for epigenetic regulation in both normal physiology and disease.

eLife Assessment

This valuable study presents a framework for a shareable data analysis pipeline aimed at improving reproducibility in neuroscience. The evidence for robustness and inter-laboratory operability is convincing. However, aspects such as accessibility for new users, flexibility for custom analyses, and plans for long-term maintenance remain incomplete. Overall, this work will be of interest to neuroscientists engaged in the analysis of large-scale neuronal recordings.

eLife Assessment

This study presents valuable findings on the physiological and computational underpinnings of the accumulation of intermittent glimpses of sensory evidence. While the authors present solid evidence to support their claims, a more exhaustive characterisation of how the different signals interact could further strengthen their case. The work will be of interest to cognitive and systems neuroscientists working on decision-making

eLife Assessment

This study uses a valuable combination of functional magnetic resonance imaging and electroencephalography (EEG) to study brain activity related to prediction errors in relation to both sensorimotor and more complex cognitive functions. It provides incomplete evidence to suggest that prediction error minimisation drives brain activity across both types of processing and that elevated inter-regional functional coupling along a superior-inferior axis is associated with high prediction error, whereas coupling along a posterior-anterior axis is associated with low prediction error. The manuscript will be of interest to neuroscientists working on predictive coding and decision-making, but would benefit from more precise localisation of EEG sources and more rigorous statistical controls.

eLife Assessment

This important study concerns the propagation of waves in bacterial biofilms, bridging active matter physics and bacterial biophysics. While the experimental observations are solid, the theoretical interpretation and model validation are currently incomplete and require further refinement. This work will be of interest to microbiologists, biophysicists, and researchers studying collective behavior in biological systems.

eLife Assessment

This paper reports a useful low-cost platform for studying mosquito behaviors such as flight activity, sugar feeding, and host-seeking responses over the course of several weeks, and demonstrates key applications of this platform. While the authors provide a biological proof of principle, the evidence that supports the validation of the tracking algorithm is incomplete; it lacks biological replicates, independent confirmation of the tracking algorithm, and data on mosquito survival.

eLife Assessment

This study uses the Drosophila mushroom body as a model to understand the molecular machinery that controls the temporal specification of neuronal cell types. With convincing experimental evidence, the authors made fundamental findings that the Pipsqueak domain-containing transcription factor Eip93F is central to the specification of a later-born neuronal subtype and in inhibiting gene expression for earlier subtypes.

eLife Assessment

This valuable study presents findings on the developmental roles of Nup107, a key nucleoporin, in regulating the larval-to-pupal transition in Drosophila melanogaster through its involvement in ecdysone signaling. The evidence supporting the authors' claims is solid, with robust experimental approaches including RNAi knockdown and rescue experiments. The authors propose that Nup107 influences EcR localization indirectly by reducing the expression of Halloween genes, a consequence of impaired Torso signaling. However, it remains uncertain whether Torso is the sole receptor tyrosine kinase involved, and this disruption ultimately leads to decreased ecdysone production. In addition, finding a mechanism would strengthen the findings as the currently proposed mechanism is not completely supported by the data.

eLife Assessment

This study employed state-of-the-art quantitative imaging and genomics approaches to address a fundamental question regarding the establishment of Polycomb domains during Drosophila embryogenesis. The critical developmental stage was pinpointed to the maternal-to-zygotic transition, rather than earlier stages, providing clarification for the field. The roles of two factors, Zelda and GAGA-factor, were investigated, which reveal that Zelda, but not GAGA-factor, contributes to this process. These compelling findings have implications for chromatin and developmental biology.

eLife Assessment

This work presents a valuable resource combining scRNA-seq and spatial transcriptomics studies to map mouse pre-clinical models of colorectal cancer, identifying distinct cellular programs and microenvironments that could enhance patient stratification and therapeutic approaches in colorectal cancer. While the evidence provided in the manuscript are not fully validated, these solid data were collected and analyzed using a validated methodology that will be of interest to the community in future studies.

eLife Assessment

In this valuable study, Taber et al. used a battery of biophysical and structural approaches to characterize the impact of erythrocytosis-related mutations in prolyl hydroxylase domain protein 2 (PHD2). The authors show that PHD2 mutant proteins are destabilized, thus supporting the tenet that dysregulation of PHD2/hypoxia induced factor (HIF) axis underpins erythrocytosis, while providing solid evidence that N-terminal ODD prolyl hydroxylation of HIF is indispensable for these phenotypes. These findings were found to be of interest for researchers focusing on oxygen sensing in homeostasis and pathological states.

eLife Assessment

This study presents a valuable and interesting finding that a combination of arginine methyltransferase inhibitors synergize with PARP inhibitors to eliminate ovarian and triple negative cancer cell lines in vitro and in vivo using preclinical mouse models. The data were collected and analyzed using solid and validated methodology and can be used as a starting point for the development of novel therapeutics. The work will be of broad interest to scientists working in the field of breast cancer and ovarian cancer.

eLife Assessment

This valuable study provides a comprehensive description of the Nematostella vectensis matrisome - the genes encoding the proteins of the extracellular matrix. The authors combine new mass spectrometry data with bioinformatic analyses of previously published genomic and single-cell RNAseq data. The analysis is thorough, and the discussion and conclusions are convincing. This work will be of interest to biologists working on the evolution of the matrisome, as well as more broadly those working with non-bilaterian animals.

eLife Assessment

This fundamental work demonstrates that compartmentalized cellular metabolism is a dominant input into cell size control in a variety of mammalian cell types and in Drosophila. The authors show that increased pyruvate import into the mitochondria in liver-like cells and in primary hepatocytes drives gluconeogenesis but reduces cellular amino acid production, suppressing protein synthesis. The evidence supporting the conclusions is compelling, with a variety of genetic and pharmacologic assays rigorously testing each step of the proposed mechanism. This work will be of interest to cell biologists, physiologists, and researchers interested in cell metabolism, and is significant because stem cells and many cancers exhibit metabolic rewiring of pyruvate metabolism.

eLife Assessment

This study presents a screen for small-molecule activators of the kinase GCN2 that phosphorylates the eukaryotic translation initiation factor 2 alpha (eIF2α) in response to diverse stress stimuli. Among the compounds identified, one stands out as a potent activator that functions independently of GCN1, which is important for probing mechanisms of Integrated Stress Response regulation and may have translational relevance in the context of pathogenic GCN2 mutations. While some reviewers found the biochemical analyses convincing, others viewed the cellular evidence as limited, particularly with respect to time points, endogenous readouts, and broader cell-type validation, which prevents a clear assessment of the compound's potential potency in a physiological context.

eLife Assessment

This fundamental work substantially advances our understanding of tissue deformation and growth patterns during the earliest stages of mammalian heart development. One of the strengths of the work is the compelling quantitative approach to analyzing time-lapse imaging data using an original computational pipeline, which goes beyond the current state of the art and provides new insights into heart tube formation. Overall, this rigorous study will be of broad interest to computational and developmental biologists studying tissue dynamics.

eLife Assessment

This work presents valuable new data on the role of D-Serine and how it competes with its stereoisomer L-Serine to influence metabolism. The work presents a variety of solid experimental data combined with simulated results to investigate the mechanisms focused on one-carbon metabolism, which is relevant for several research fields. However, some claims are only partially supported by data, and critical areas comparing L- vs D-Serine and further mechanistic studies are incomplete. Furthermore, while the work has potential for various fields, the work has only been studied in a limited cell type and context.

eLife Assessment

Plasmodesmata are channels that allow cell-cell communication in plants; based on the functional similarities between facilitated transport within plasmodesmata and into the nucleus, the authors speculate that nuclear pore complex proteins might be involved in plasmodesmata function. If supported, this would transform our understanding of cell-to-cell communication in plants. The authors localize nuclear pore complex proteins to plasmodesmata using proteomics and heterologous overexpression; however, the data are incomplete since key controls for localization, functionality, and expression level of fluorescent protein fusions are absent.

eLife Assessment

This important study demonstrates the potential of synthetic gene circuits to detect and target aberrant RAS activity in cancer cell lines. The circuit design is novel and the evidence supporting the claims is convincing. As a proof-of-concept, this will be of broad interest to researchers in synthetic biology and therapeutics development, while future work will be required to help translate this technology toward clinical applications in cancer therapeutics and address potential limitations of the strategy.

eLife Assessment

This study constitutes a fundamental advance for the uveal melanoma research field that might be exploited to target this deadly cancer and, more generally, for targeting transcriptional dependency in cancers. This work substantially advances our understanding of pharmacological inhibition of SWI/SNF as a therapeutic approach for cancer. The study is well written and provides compelling evidence, including comprehensive datasets, compound screens, gene expression analysis, epigenetics, as well as animal studies.

eLife Assessment

The is a valuable evaluation of a previously published simulation model on the role of heterozygote advantage in shaping MHC diversity, showing that the conclusions from this model hold only within a narrow parameter range that might be unrealistic. The author presents an alternative model, in which MHC homozygotes with duplicated MHC genes outperform heterozygotes with single genes, thereby challenging the explanation that heterozygote advantage will lead to high allelic variation at a given MHC gene. The topic is highly relevant for eco-immunology and evolutionary genetics, but several major aspects of the author's claim need to be clarified to make the model interpretable. While the work has the potential to improve our understanding of the question of how the extraordinary diversity at the MHC locus evolves, without this addition, the conclusions remain incomplete.

eLife Assessment

This valuable study reveals that the GSK-3 inhibitor AZD2858 inhibits the formation of TOPBP1 condensates and hence DNA damage responses in colorectal cancer cells. The evidence supporting the claims of the authors is convincing, although uncovering how this drug blocks bio-condensate formation would have strengthened the study. The work will be of interest to cancer researchers searching for synergistic drug combination strategies.

[Editors' note: this paper was reviewed by Review Commons.]

eLife Assessment

This study reports the important finding that the dynamin inhibitor Dyngo-4a broadly affects lipid packing and plasma membrane dynamics, independently of its action on dynamin. While solid computational, biophysical, and cell-based evidence supports this conclusion, there is incomplete support for the authors' main claim on the role of lipid packing in caveolae internalization, as the causal relationship remains unclear and direct analyses are lacking. With stronger evidence, this work would be of significant interest to cell biologists, biophysicists, and chemists interested in membrane remodeling and drug-membrane interactions.

[Editors' note: this paper was reviewed by Review Commons.]

eLife Assessment

This manuscript describes a novel method for determining the mechanical parameters of the kinesin, KIF1A, that uses fluorescence microscopy and does not require an optical tweezer apparatus. The length of a tethered fluorescent DNA nanospring is measured as the kinesin moves processively along the microtubule and then stalls. The work reports important findings, and (barring a few exceptions) the evidence supporting the claims is generally convincing.

eLife Assessment

This paper presents valuable findings on how autophagosomes are positioned along microtubules for their efficient fusion with lysosomes, providing significant insights into the mechanism. The evidence supporting the conclusions is solid, with high-quality fluorescence microscopy combined with Drosophila genetics. This work will be of broad interest to cell biologists interested in autophagy and related cell biology fields.

eLife Assessment

This is an important study that utilizes proteomic and genetic approaches to identify the glycoprotein quality control factor malectin as a pro-viral host protein involved in the replication of coronavirus. The evidence supporting this conclusion is convincing, although continued elucidation of the mechanistic basis of malectin-mediated viral replication would further strengthen these findings. This work will be of interest to cell biologists studying the molecular mechanisms of glycoprotein quality control and virologists studying the host-pathogen interactions.

eLife Assessment

The paper is a fundamental study examining the role of CDK12 loss in prostate cancer. While previous studies have suggested that CDK12 loss confers HRD phenotypes, clinical trials using PARPi in CDK12 altered patients have not demonstrated significant benefit. This work investigates these mechanisms in depth and provides compelling evidence. A comprehensive genomic analysis serves an excellent resource to the field, showing that biallelic CDK12 alterations do not have genomic features of HRd. Moreover, the study explored both acute and chronic deletion of CDK12, with data suggestive of CDK12-altered cells being uniquely sensitive to CDK13 inhibition. While some minor weaknesses have been previously noted by the reviewers, the authors have adequately addressed these concerns with appropriate rigor.

eLife Assessment

This useful study informs the transcriptional mechanisms that promote stem cell differentiation and prevent degeneration in the adult eye. Through inducible mouse mutagenesis, the authors uncover a dual role for a transcription factor (Sox9) in stem cell differentiation and prevention of retinal degeneration. The data at hand convincingly support to the main conclusions. The study will be of general interest to the fields of neuronal development and neurodegeneration.

eLife Assessment

Shen et al. present a computational account of individual differences in mouse exploration when faced with a novel object in an open field from a previously published study (Akiti et al.) that relates subject-specific intrinsic exploration and caution about potential hazards to the spectrum of behaviors observed in this setting. Overall, this computational study is an important contribution that leverages a very general modeling framework (a Bayes Adaptive Markov Decision Process) to quantify and interrogate distinct drivers of exploratory behavior under potential threat. Given their assumptions, the modeling results are convincing: the authors are able to describe a substantial amount of the behavioral features and idiosyncracies in this dataset, and their model affords a normative interpretation related to inherent risk aversion and predation hazard "flexibility" of individual animals and should be of broad interest to researchers working to understand open-ended exploratory behaviors.

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

This manuscript reports valuable results on the role of MDC1 and Treacle in DSB repair in rDNA repeats. It has been previously established that MDC1 is replaced by Treacle as the main adaptor in the nucleolar DNA damage response. This work provides convincing evidence that MDC1 is required for the recruitment of RAD51 and BRCA1 to DSBs in rDNA. The work involves multiple MDC1 knockout models and establishes that RFN8-RNF168 act downstream of MDC1 in the recruitment of the HR machinery to nucleolar DSBs.

eLife Assessment

In this important work, the authors present a new transformer-based neural network designed to isolate and quantify higher-order epistasis in protein sequences. They provide solid evidence that higher-order epistasis can play key roles in protein function. This work will be of interest to the communities interested in modeling biological sequence data and understanding mutational effects.

eLife Assessment

This manuscript uses simulations to describe the dynamics of the Pseudomonas-derived cephalosporinase PDC-3 β-lactamase and its mutants to better understand antibiotic resistance. The finding that clinically observed mutations alter the flexibility of the Ω- and R2-loops, reshaping the cavity of the active site, is useful to the field. However, the evidence is considered incomplete; there is a lack of description of methods, and there is a need for additional analysis to demonstrate statistical significance, visualisation of the Markov states, analysis to explain changes due to the different mutations, and possible simulations in the presence of substrates to shed direct light on modulation mechanisms.

eLife Assessment

This manuscript employs cryo-EM, mutational analysis, and biochemical assays to explore the molecular basis by which glutamine promotes filamentation and regulates the activity of human glutamine synthetase (hGS) by stabilizing interactions between hGS decamers. The studies supporting this mechanism are solid, but could be improved by providing more clarity and by addressing methodological issues in the cryoEM data processing workflow. This work will be of particular interest and useful to groups interested in understanding the molecular basis of nutrient sensing, cellular metabolism, and structural regulation of enzyme activity.