eLife assessment

This valuable study asks how Promyelocytic leukemia protein (PML) becomes associated with the nucleoli of cells (PML Nucleolar Associations, PNAs) upon various genotoxic stimuli. Using immunostaining analysis with induced DNA double-strand breaks (DSBs) in rDNA repeats, the authors provide solid evidence that PNAs are triggered mostly by the inhibition of topoisomerase and RNA polymerase I, which is augmented by homologous recombination but not by the non-homologous end joining double-strand break repair pathway. The findings have potential implications for a better understanding of how DNA damage in ribosomal DNA is repaired for genome stability. This paper is of interest to researchers in the fields of nuclear structure and DNA repair.

eLife assessment

This valuable study examines the activity and function of dorsomedial striatal neurons in estimating time. The authors used various causal and correlational techniques to investigate how these pathways collectively contribute to interval timing in mice and found that the direct and indirect striatal pathways perform opposing roles in processing elapsed time. The evidence is solid. The manuscript would interest neuroscientists examining how striatum contributes to behavior.

eLife assessment

This valuable study introduces SCellBOW, a novel tool leveraging natural language processing techniques to enhance cell clustering and infer survival risks from single-cell RNA sequencing data. The methodology and results are convincing, demonstrating superior clustering performance and the ability to assign risk scores to cancer cell clusters across multiple datasets. SCellBOW's unique approach promises significant advancements in understanding cancer cell heterogeneity and identifying aggressive cancer cell subgroups.

eLife assessment

This valuable study, which seeks to identify factors from the glial niche that support and maintain neural stem cells, reports a novel role for ferritin in this process. The authors provide solid evidence that defects in larval brain development in Drosophila, resulting from ferritin knockdown, can be attributed to impaired Fe-S cluster activity and ATP production. The findings of this well-conducted study will be of interest to oncologists and neurobiologists.

eLife assessment:

This useful study shows that the essential Acinetobacter baumannii gene Aeg1 likely plays an key role in cell division. The strength of the work is the discovery that the depletion of Aeg1 leads to cell filamentation and that gain-of-function mutations in cell division genes FtsB and FtsL rescue the lethality of Aeg1 depletion. However, Aeg1's localization pattern and its requirement for other division proteins' localizations require further characterization of the functionality of fluorescent fusion proteins, fluorescence images of higher quality, and improvements in statistic qualifications, leaving the study' evidence for Aeg1's exact role in cell division incomplete at this time. In conclusion, the critical role of Aeg1 in the assembly of the A. baumannii divisome has yet to be established unambiguously.

eLife assessment

Working with a diverse panel of field-grown rice accessions, this valuable study measures changes in transcript abundance, tests for patterns of selection on gene expression, and maps the genetic basis of variation in gene expression in normal and high salinity conditions. The authors provide solid evidence that salinity treatment increases the number of genes with mean expression levels away from the optimum, and that a relatively small number of genes are hotspots for genetic variants that affect genome-wide patterns of variation in gene expression under high salinity. The design, clarity, and interpretation of several statistical analyses can be improved, additional opportunities for integration among datasets and analyses could be realized, and genetic manipulation would be required to confirm the functional involvement of any specific genes in regulatory networks or organismal traits that confer adaptation to higher salinity conditions. The manuscript will not only be of interest to evolutionary biologists studying the genetics of complex traits, but it will also be a resource for plant biologists studying mechanisms of abiotic stress tolerance.

eLife assessment

This study provides new insights into the expression profile of ILCs that demonstrate a history of RAG expression. It examines in part the potential intrinsic regulation of RAG expression and seeks to understand how the epigenetic state of ILCs is established, although a full understanding of intrinsic factors is incomplete. The work provides an important molecular dataset, and with further strengthening of the understanding of intrinsic regulation, this paper would be of interest more broadly to cell biologists seeking to understand immune cell development.

eLife assessment

This study provides a valuable characterization of individual sarcomere's contractility and synchrony in spontaneously beating cardiomyocytes as a function of substrate stiffness. The authors, however, provide an incomplete explanation for the observed heterogeneous and stochastic dynamics, so that the work remains mainly descriptive. The work will be of interest to scientists working on muscle biophysics, nonlinear dynamics, and synchronization phenomena in biological systems.

eLife assessment

This study demonstrates a novel role for SIRT4; a mitochondrial deacetylase, shown to translocate into nuclei where it regulates RNA alternative splicing by modulating U2AF2 and the gene expression of CCN2 in tubular cells in response to TGF-β. This fundamental work substantially advances our understanding of kidney fibrosis development and offers a potential therapeutic approach. The evidence supporting the conclusions of a SIRT4-U2AF2-CCN2 axis activated by TGF-β is compelling and adds a new layer of complexity to the pathogenesis of chronic kidney disease.

eLife assessment

Zhao et al. report valuable adverse effects on cell proliferation, differentiation and gene expression, possibly linked to reduced binding activity of the transcription factor GTF2IRD1 to the transthyretin (TTR) promoter, in a human forebrain organoid model of Williams Syndrome (WS). The authors provide incomplete evidence of the effects of GTF2IRD1, a mutated gene in WS, on altering MAPK/ERK pathway activity, a well-recognized target in cell proliferation.

eLife assessment

The authors present a potentially useful approach of broad interest arguing that anterior cingulate cortex (ACC) tracks option values in decisions involving delayed rewards. The authors introduce the idea of a resource-based cognitive effort signal in ACC ensembles and link ACC theta oscillations to a resistance-based strategy. The evidence supporting these new ideas is incomplete and would benefit from additional detail and more rigorous analyses and computational methods.

eLife assessment

This important study builds on a previous publication, demonstrating that T. brucei has a continuous endomembrane system, which probably facilitates high rates of endocytosis. Using a range of cutting-edge approaches, the authors present compelling evidence that an actomyosin system, with the myosin TbMyo1 as an active molecular motor, is localized close to and can associate with the endosomal system in the bloodstream form of Trypanosoma brucei. It shows convincingly that both actin and Myo I play a role in the organization and integrity of the endosomal system: both RNAi-mediated depletion of Myo1, and treatment of the cells with latrunculin A resulted in endomembrane disruption. This work should be of interest to cell biologists and microbiologists working on the cytoskeleton, and unicellular eukaryotes.

eLife assessment

This is a useful study on sex differences in gene expression across organs of four mice taxa, although there are some shortcomings in the data analyses and interpretations that should to be better placed in the broader context of the current literature. Hence, the evidence in the current form is incomplete, with several overstated key conclusions.

eLife assessment

This paper provides valuable findings related to the impact and timing of exogenous interleukin 2 on the balance of exhausted (Tex) versus effector (Teff) that differentiate from precursors T cells (Tpex) during chronic viral infection. While the data appear solid, the overall claims that IL-2 suppresses Tpex are only partially supported, with the rationale for the timing of IL-2 treatment and its underlying mechanisms remaining unclear.

eLife assessment

This is a mechanistic study showing the effect of combining inhibition of autophagy (through ULK1/2) and KRAS (using sotorasib) on KRAS mutant NSCLC making the study valuable to cancer biologists and more broadly in a clinical setting. The evidence generated by GEM mouse models and cell lines is solid but could be further strengthened by increasing the mouse cohort size. This study holds translational relevance beyond NSCLC to other indications that carry KRAS mutations.

eLife assessment

This is a useful manuscript describing the competitive binding between Parkin domains to define the importance of dimerization in the mechanism of Parkin regulation and catalytic activity. The evidence supporting the importance of Parkin dimerization for an 'in trans' model of Parkin activity described in this manuscript is solid, but lacks more stringent and biochemical characterization of competitive binding that could provide more direct evidence to support the author's conclusions. This work will be of interest to those focused on defining the molecular mechanisms involved in ubiquitin ligase interactions, PINK-Parkin-mediated mitophagy, and mitochondrial organellar quality control.

eLife assessment

This study presents important findings on the different polymorphs of alpha-synuclein filaments that form at various pH's during in vitro assembly reactions with purified recombinant protein. Of particular note is the discovery of two new polymorphs (1M and 5A) that form in PBS buffer at pH 7. The strength of the evidence presented is convincing. The work will be of interest to biochemists and biophysicists working on protein aggregation and amyloids.

eLife assessment

Using multiple public datasets, this study investigates associations between retrotransposon element expression and methylation with age and inflammation. The study is valuable because a systematic analysis of retrotransposon element expression during human aging has been lacking, but the provided data must be considered incomplete due to the sole reliance on microarray expression data for the core analyses.

eLife assessment

This is a solid study that follows a well-established canvas for variant-to-gene prioritisation using 3D genomics, applying it to activated T cells. The authors go some way in validating the lists of candidate genes, as well as exploring the regulatory architecture of a candidate GWAS locus. Jointly with data from previous studies performing variant-to-gene assignment in activated CD4 T cells (and other immune cells), this work provides a useful additional resource for interpreting autoimmune disease-associated genetic variation.

eLife assessment

This valuable study demonstrates that AARS2 is crucial for protecting cardiomyocytes from ischemic stress by shifting energy metabolism towards glycolysis through PKM2, presenting a novel therapeutic target for myocardial infarction. The findings are supported by solid evidence, including cardiomyocyte-specific genetic modifications, functional assays, and ribosome profiling, which together robustly validate the AARS2-PKM2 signaling pathway's role in cardiac protection.

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, although the conclusions regarding the effect of phosphorylation would be strengthened by quantification of the phosphopeptides. Recent work on the role of APC7 in APC/C activity in neurones should also be discussed with respect to the mode of action of the APC/C in human versus budding yeast cells.

eLife assessment

This study assessed the virulence and immune responses of different M. tuberculosis lineages using a 3D in vitro granuloma model. The useful findings support the functional impact of M. tuberculosis natural diversity on host-pathogen interactions but are incomplete and only partially support the claims. The study will interest researchers working on mycobacteria and understanding how genetic diversity influences virulence and immunity outcomes.

eLife assessment

This valuable paper presents convincing evidence that changing the constraint of how long to stop at an intermediate target significantly influences the degree of coarticulation of two sequential reaching movements, as well as their response to mechanical perturbations. Using an optimal-control framework, the authors offer a normative explanation of how both co-articulated and separated sequential movement can be understood as an optimal solution to the task requirements.

eLife assessment

This is an important study that describes an elegant modelling driven approach to design of allosteric antagonists for CXCR4 that have a selective effect on receptor nanocluster formation, cell polarisation and chemotaxis, but spare binding of CXCL12 to the receptor and inhibition of adenylate cyclase. This enables selective targeting of processes dependent upon cell polarisation and chemotaxis without impacting signalling effects and may avoid some of the toxicity associated with antagonists that target CXCL12 binding and thus block all CXCR4 signalling. The revised manuscript offers convincing evidence to support the claims. The modelling work is better described and additional data has been presented that better illustrates the unique features of the new antagonist. The in vivo studies in the zebrafish model open a path to studies in mammalian models.

eLife assessment

This paper presents important computational modeling work that provides a mechanistic account for how memory representations become integrated or differentiated (i.e., having distinct neural representations despite being similar in content). The authors provide convincing evidence that simple unsupervised learning in a neural network model, which critically weakens connections of units that are moderately activated by multiple memories, can account for three empirical findings of differentiation in the literature. The paper also provides insightful discussion on the factors contributing to differentiation as opposed to integration, and makes new predictions for future empirical work.

eLife assessment

ImmCellTyper presents a useful toolkit for CyTOF data analysis, integrating BinaryClust for semi-supervised clustering and cell type annotation. The evidence supporting the findings is convincing, with appropriate and validated methodology. This tool will be helpful to researchers in immunology and cytometry, offering a robust solution for cell type identification and differential analysis.

eLife assessment

This valuable study investigates how the human brain flexibly adjusts its representations of the world as the environment continually changes. It utilizes a unique dataset in which participants view thousands of natural scenes across many fMRI sessions over multiple months. The evidence supporting the claims of the authors is incomplete, with statistical inference not always warranted. The study would interest a broad readership in cognitive neuroscience.

eLife assessment

This joint computational/experimental study demonstrates the ability of synthetic peptides derived from the stalk-tethered agonist in Polycystin-1 (PC1) to re-activate signaling by a stalkless C-terminal fragment of PC1. The study is valuable as it discovered peptide agonists for PC1 and the integrated in vitro and in silico approach is potentially applicable to the analysis of related systems. Following the revision, the line of evidence presented in the current manuscript is considered convincing.

eLife assessment

This valuable study analyzes the role of rpgrip1l encoding a ciliary transition zone component in the development of neuroinflammation and scoliotic phenotypes in zebrafish. Through proteomic and experimental validation in vivo, the authors demonstrated increased Annexin A2 expression and astrogliosis in the brains of scoliosis fish. Anti-inflammatory drug treatment restored normal spine development in these mutant fish, thus providing additional convincing evidence for the role of neuroinflammation in the development of scoliosis in zebrafish.

eLife assessment

This important work describes a compelling analysis of DNA damage-induced changes in nascent RNA transcripts, and a genome-wide screening effort to identify the responsible proteins. A significant discovery is the inability of arrested cells to undergo DNA damage-induced gene silencing, which, is attributed to an inability to mediate ATM-induced transcriptional repression. This work will be of general interest to the DNA damage, repair, and transcription fields, with a potential impact on the cancer field.

eLife assessment

This fundamental work substantially advances our understanding of the role of calcium-binding proteins 1 and 2 (CaBP1 and CaBP2) for generating sustained calcium currents in mouse inner hair cells and their capacity for indefatigable exocytosis. The evidence supporting the conclusions is compelling, with rigorous in vitro and in vivo physiological experiments and state-of-the-art microscopy. The work will be of broad interest to synaptic physiologists, cellular biochemists, and hearing researchers.

eLife assessment

The fundamental findings of this work substantially advance our understanding of the impact of the host on its gut microbes. The authors provided compelling evidence at single-cell resolution that the host can drive heterogeneity in the populations of gut microbes with significant consequences for the host physiology.

eLife assessment

In this valuable study, the authors sought to investigate the associations of age at breast cancer onset with the incidence of myocardial infarction and heart failure. Based on results from a series of compelling statistical analyses, the authors conclude that a younger onset age of breast cancer is associated with myocardial infarction and heart failure, highlighting the need to carefully monitor the cardiovascular status of women who have been diagnosed with breast cancer.

eLife assessment

Cav2 voltage-gated calcium channels play key roles in regulating synaptic strength and plasticity. In contrast to mammals, invertebrates like Drosophila encode a single Cav2 channel, raising questions on how diversity in Cav2 is achieved from a single gene. Here, the authors present convincing evidence that two alternatively spliced isoforms of the Cac gene (cacophony, also known as Dmca1A and nightblindA) enable diverse changes in Cav2 expression, localization, and function in synaptic transmission and plasticity. These valuable findings will be of interest to a variety of researchers.

eLife assessment

Guan and colleagues present solid arguments to address the question of how a single neural stem cell produces a defined number of progeny, and what influences its decommissioning. The focus of the experiments are two well-studied RNA-binding proteins: Imp and Syp. This is valuable work that will be of interest to the scientific community.

eLife assessment

This study by Nandy and colleagues examined relationships between behavioral state, neural activity, and trial-by-trial variability in the ability to detect weak visual stimuli. They present useful findings indicating that certain changes in arousal and eye-position stability, along with patterns of synchrony in the activity of neurons in different layers of cortical area V4, can show modest correspondences to changes in the ability to correctly detect a stimulus. At present, however, the findings are based on data and analyses that are somewhat incomplete but could be improved with further revisions.

eLife assessment

This paper proposes a valuable new method for the assessment of the mean kurtosis for diffusional kurtosis imaging by utilizing a recently introduced sub-diffusion model. The evidence supporting the claims that this technique is robust and accurate in brain imaging is solid; however, there is a need to include a summary of the clear limitations.

eLife assessment

In this useful study, the authors report the efficacy, hematological effects, and inflammatory response of the BPaL regimen (containing bedaquiline, pretomanid, and linezolid) compared to a variation in which Linezolid is replaced with the preclinical development candidate spectinamide 1599, administered by inhalation in tuberculosis-infected mice. The authors provide convincing evidence that supports the replacement of Linezolid in the current standard of care for drug-resistant tuberculosis. The work will be of interest to those studying tuberculosis treatment regimens.

eLife assessment

This study addresses a question in sensory ethology and active sensing in particular. It links the production of a specific signal - electrosensory chirps - to various contexts and conditions to argue that the main function is to enhance conspecific localization rather than communication as previously believed. The study provides a lot of valuable data, but the methods section is incomplete making it difficult to evaluate the claims.

eLife assessment

This important study correlates the size of various prefrontal brain regions in primate species with socioecological variables like foraging distance and population density. The evidence presented is solid but the approach and conclusions are limited to primates with well-defined gyri.

eLife assessment

This manuscript describes a creative approach using dual-component gRNAs to create a new class of molecular proximity sensors for genome editing. The authors demonstrate that this tool can be coupled with several different gene editing effectors, and the authors convincingly show that this functions as designed. This important study represents this first-of-its kind technology with key baseline activity metrics ready for future developmental approaches.

eLife assessment

The study is a valuable contribution to the question of evolutionary shifts in neuronal proliferation patterns and the timing of developmental progressions. The authors present solid support for the presence of type-II NB lineages in the beetle Tribolium with the same molecular characteristics as the counterparts in the fly Drosophila, but differences in lineage size and number. While presenting a number of interesting observations, further evidence will be required to show that the observed differences are indeed responsible for the differences in developmental timing of the central complex in the two insect species.

eLife assessment

This is a valuable study that provides CCR7-APEX2 proximity labelling mass spectrometry data that is expected to provide new insights into CCR7 signalling partners and pathways. The study is technically solid and easy to follow, however, there are some concerns that many of the highlighted findings are repetitive of prior work and that this is not clearly acknowledged. It would increase the impact of the study if the confirmatory nature of some findings were acknowledged. This is of value to the community, and there are likely multiple opportunities to use the APEX2 data set to extend these findings, strengthen some claims, and even explore a new pathway identified in the APEX2 data set.

eLife assessment

This is a valuable study providing solid evidence that the Mediator kinase module mediates an elevated inflammatory response, manifested by heightened cytokine levels, associated with Downs syndrome (DS) via transcriptional changes impacting cell signaling and metabolism, which has significance for the treatment of DS and other chronic inflammatory conditions. Particular strengths of the study include the combined experimental approaches of transcriptomics, untargeted metabolomics, cytokine screens, and the use of sibling-matched cell lines (trisomy 21 vs disomy 21) from various donors. Less certain is that the Mediator kinase plays a meaningful role in regulating mRNA splicing. Further evidence that nuclear receptors are activated by changes in lipid levels and that mitochondrial function is substantially reduced on Mediator kinase inhibition would strengthen the work.

eLife assessment

This manuscript presents valuable findings on two isolates of deep sea Lentisphaerae strains, which further our understanding of deep sea microbial life. The manuscript's primary claim is that phage isolates augment polysaccharide use in Pseudomonas bacteria, with preliminary evidence for the potential auxiliary metabolic genes in chronic phage infection and/or host proliferation. The strength of the evidence is overall solid and there are only minor weaknesses regarding the mechanism of polysaccharide use by the phages and the evidence for chronic infection. Overall, the data on Lentisphaerae strains will deepen our understanding of microbial life in the deep sea.

eLife assessment

This important study provides a new perspective on why preparatory activity occurs before the onset of movement. The authors report that when there is a cost on the inputs, the optimal inputs should start before the desired network output for a wide variety of recurrent networks. The authors present compelling evidence by combining mathematically tractable analyses in linear networks and numerical simulation in nonlinear networks.

eLife assessment

This is a potentially useful study that shows changes in the chromatin landscape of GABAergic neurons in induced pluripotent stem cells (iPSCs) derived from both Dravet Syndrome (DS) patients and healthy donors. The strength of the evidence is currently incomplete because the authors compared iPSCs from different individuals, rather than isogenic controls. A strategy for minimizing variability across cell lines is used, but the explanation is not complete. The revised manuscript adds RNAseq and qPCR measurements of the expression of the gene SCN1A, however these do not appear to agree, perhaps because of the way the qPCR measurements are normalized, and there is no measurement of Nav1.1, the gene product thought to be responsible for the majority of DS cases. Hence the evidence that there is reduced expression of SCN1A or its gene product is not complete and therefore it is difficult to evaluate whether or not the observed epigenetic changes are causal. The work would potentially be of interest to scientists who study development, developmental disorders, and epigenetic contributions to disease.

eLife assessment

This important study identifies the anti-inflammatory function of PEGylated PDZ peptides that are derived from the ZO-1 protein. Results from cellular and in vivo experiments tracking key inflammatory markers are compelling. Although the present study would benefit from investigating chronic inflammation conditions using microbe and protein data, the work provides a proof of concept for developing novel strategies against acute inflammatory conditions such as sepsis.

eLife assessment

This important study presents a novel pipeline for the large-scale genomic prediction of members of the non-ribosomal peptide group of pyoverdines based on a dataset from nearly 2000 Pseudomonas genomes. The advance presented in this study is based on convincing evidence. This study of bacterial siderophores has broad theoretical and practical implications beyond a singular subfield.

eLife assessment

This valuable study uses neuroanatomical techniques to investigate somatosensory projections from the elephant trunk to the brainstem. Given its unique specializations, understanding how the elephant trunk is represented within the brain is of general interest to evolutionary and comparative neuroscientists. The authors present solid evidence for the existence of a novel isomorphism in which the folds of the trunk are mapped onto the trigeminal nucleus; however, due to their unusual structure, some uncertainty remains about the identification and anatomical organization of nuclei within the elephant brainstem.

eLife assessment

This fundamental study provides insights into how pathogens respond, on a systemic level including several gene targets and clusters, to selected antimicrobial molecules. Compelling evidence is provided, through multi-omics and functional approaches, that very similar molecules originally designed to target the same bacterial protein act differently within the context of the whole set of cellular transcripts, expressed proteins, and pre-lethal metabolic changes. Given the rapid accumulation of omics data and the much slower capacity of extracting biologically relevant insights from big data, this work exemplifies how the development of sensitive data analysis is still a major necessity in modern research.

eLife assessment

This important paper addresses the process by which cryptic splice sites that occur randomly in exons are ignored by the splicing machinery. Integrating state-of- the-art genome-wide approaches such as CLIP-seq with the study of individual examples, this study convincingly implicates members of RBMX family of RNA binding proteins in such cryptic splice site suppression and showcases its importance for the fidelity of expression of genes with very large exons.

eLife assessment

This study presents a valuable finding that the blood-brain barrier functionality changes with age and differs between males and females. The analysis is solid, comprising a large and racially diverse dataset, and utilizes a contrast-agent-free MRI method. Since limited work has been done in the MRI field on the blood-brain barrier using this method, this study is of great interest to neuroimaging researchers and clinicians.

eLife assessment

Peng et al. reported important findings that 36THz high-frequency terahertz stimulation (HFTS) could suppress the activity of pyramidal neurons by enhancing the conductance of voltage-gated potassium channels. The significance of the findings in this paper is that chronic pain remains a significant medical problem, and there is a need to find non-pharmacological interventions for treatment. The authors present convincing evidence that high-frequency stimulation of the anterior cingulate cortex can alter neuronal activity and improve sensory pain behaviors in mice.

eLife assessment

This important research describes the sensory innervation of oral tumors, with potential implications for understanding cancer-induced alterations in motivation and anhedonia in a mouse model. These findings are solid and are supported by anatomical and transcriptional changes in the tumor that suggest sensory innervation, neural tracing, and neural activity measurements. While nerve innervation of the tumor and associated increase in brain activity is well-supported, future studies could enhance specificity by employing more targeted genetic and pharmacological tools to manipulate these circuits selectively.

eLife assessment

This important study reports findings on the GnRH pulse generator's role in androgen-exposed mouse models, providing further insights into PCOS pathophysiology and advancing the field of reproductive endocrinology. The experimental data were collected using cutting-edge methodologies and were solid. However, it is noteworthy that the findings, while interesting, are primarily applicable to mouse models, and their translation to human physiology requires cautious interpretation and further validation. This work will be of interest to endocrinologists and reproductive biologists.

eLife assessment

This study provides a potentially valuable understanding of the contribution of different striatal subregions, the anterior Dorsal Lateral Striatum (aDLS) and the posterior Ventrolateral Striatum (pVLS), to auditory discrimination learning. The combined methods used to probe this are compelling, yet the data presented are incomplete to support the conclusions. There is insufficient data visualization of learning vs. performance, and missing details about timing of manipulations and microPET imaging.

eLife assessment

This important study compared the brain development trajectories of humans and macaque monkeys to quantify different evolutionary effects of convergent and divergent neural pathways between the two species. The cross-species evidence is solid, based on brain age prediction models that were carefully developed by using public MRI datasets of both humans and macaque monkeys. The findings will be of interest to neuroscientists, developmental biologists, and evolutionary biologists.

eLife assessment

This important work offers an experimental structural characterization of the Tramtrack-like BTB/POZ domains in insects, revealing that these domains form stable hexameric assemblies. The structural evidence is convincing, and validated by fold prediction and evolutionary pathway analyses. This paper would be of interest to structural and evolutionary biologists.

eLife assessment

This important study explores and delineates multivariate mappings between brain structure and functional measures with latent dimensions of psychopathology. This work provides solid evidence for the existence of such mappings and charts the relationship between different neurobiological measures and distinct dimensions of psychopathology. This work will be of broad interest within the neuroscience field.

eLife assessment

This potentially important work presents a tool for performing phylogenetic taxonomic classification of DNA sequences. In terms of methodology, the work is compelling. The authors perform a benchmark experiment against current state-of-the-art tools using real and simulated datasets to demonstrate where the novel tool stands in the context of existing methods. However, the experimentation is still incomplete. It would benefit from a more thorough exploration of existing methods as well as data sets that better represent real-world use cases.

eLife assessment

In this study, Perez-Lopez and colleagues examine an important function of the chemokine CCL28 in mucosal host defenses against the gut bacterial pathogen Salmonella Typhimurium and lung pathogen Acinetobacter baumanii. They find that CCL28-CCR3 axis regulates neutrophil recruitment and function, and promotes bacterial clearance in one infectious context but exacerbates disease against the other pathogen. Therefore, CCL28 plays a critical role in mucosal immunity and neutrophil biology that differentially affects host defenses against pathogens.

Editors Assessment:

RAD-Seq (Restriction-site-associated DNA sequencing) is a cost-effective method for single nucleotide polymorphism (SNP) discovery and genotyping. In this study the authors performed a kinship analysis and pedigree reconstruction for two different cattle breeds (Angus and Xiangxi yellow cattle). A total of 975 cattle, including 923 offspring with 24 known sires and 28 known dams, were sampled and subjected to SNP discovery and genotyping using RAD-Seq. Producing a SNP panel with 7305 SNPs capturing the maximum difference between paternal and maternal genome information, and being able to distinguish between the F1 and F2 generation with 90% accuracy. Peer review helped highlight better the practical applications of this work. The combination of the efficiency of RNA-seq and advances in kinship analysis here can helpfully help improve breed management, local resource utilization, and conservation of livestock.

This evaluation refers to version 1 of the preprint

Editors Assessment: This work is part of a series of papers from the Hong Kong Biodiversity Genomics Consortium sequencing the rich biodiversity of species in Hong Kong (see https://doi.org/10.46471/GIGABYTE_SERIES_0006). This example assembles the genome of the black-faced spoonbill (Platalea minor), an emblematic wading bird from East Asia that is classified as globally endangered by the IUCN. This Data Release reporting a 1.24Gb chromosomal-level genome assembly produced using a combination of PacBio SMRT and Omni-C scaffolding technologies. BUSCO and Merqury validation were carried out, gene models created, and peer reviewers also requested MCscan synteny analysis. This showed the genome assembly had high sequence continuity with scaffold length N50=53 Mb. Presenting data from 14 individuals this will hopefully be a useful and valuable resources for future population genomic studies aimed at better understanding spoonbill species numbers and conservation.

*This evaluation refers to version 1 of the preprint *

eLife assessment

This study provides useful in vitro evidence to support a mechanism whereby dyslipidemia could accelerate renal functional decline through the activation of the AT1R/LOX1 complex by oxLDL and AngII. As such, it improves the knowledge regarding the complex interplay between dyslipidemia and renal disease and provides a solid basis for the discovery of novel therapeutic strategies for patients with lipid disorders. The methods, data, and analyses support the presented findings, although the observed variability and need for further in vivo validation require additional research in this key area.

eLife assessment

The study explored the influence of magnesium on phenotypic antibiotic resistance in two Vibrio model bacteria. This research is fundamental for revealing the phenotypic antibiotic resistance mechanism utilized by the specified model bacteria in elevated levels of magnesium. The study produced solid evidence indicating that in high concentrations of magnesium, the efficacy of selected antibiotics was diminished due to decreased biosynthesis of unsaturated fatty acids and PE, along with an increase in the biosynthesis of PG.

eLife assessment

This valuable study demonstrates how proximity labeling with streptavidin can be used to boost fluorescence signals in otherwise hard-to-label regions of cells. The experimental verification of amplification of fluorescence near epitope tags in phase-separated compartments is solid, demonstrating enhanced signal-to-noise compared to immunofluorescence. This study will be of particular interest to those using correlative light and electron microscopy or expansion microscopy when the signal is limiting or inaccessible.

eLife assessment

The authors made an important finding that CD29+/CD56+ progenitor cells isolated from human muscles have the potential to differentiate to tendons in vitro and in vivo. The author's approach to testing the tenogenesis of the CD29+/CD56+ progenitors is solid, and the conclusion is supported by enough evidence with minor flaws. This work will be of interest to the population who need tendon regeneration from their injury.

eLife assessment

In this potentially important study, the authors conducted atomistic simulations to probe the salt-dependent phase separation of the low-complexity domain of hnRN-PA1 (A1-LCD). The authors have identified both direct and indirect mechanisms of salt modulation, provided explanations for four distinct classes of salt dependence, and proposed a model for predicting protein properties from amino acid composition. There is a range of opinions regarding the strength of evidence, with some considering the evidence as incomplete due to the limitations in the length and complexity of the atomistic MD simulations. The work should be put into a better context in relation to previous studies of salt effects on protein phase separation.

eLife assessment

This manuscript reports important findings on the impact of maternal obesity on oocyte methylation and its transgenerational effects. The evidence presented to substantiate the major claims appears incomplete. This study would be of interest to biologists in the fields of epigenetics and metabolism.

eLife assessment

This study reports the adhesion G-protein-coupled receptor A3 (ADGRA3) as a possible target for activating adaptive thermogenesis in white and brown adipose tissue. The study provides valuable insights for scientists who study metabolism, obesity, and adipose tissue biology. Meanwhile, the experimental evidence supporting the claim is incomplete, and more rigorous approaches are needed to demonstrate the relevance of this receptor in adipose tissue biology.

eLife assessment

In an important fMRI study with an elegant experimental design and rigorous cross-decoding analyses, this work shows a solid dissociation between two parietal regions in visually processing actions. Specifically, aIPL is found to be sensitive to the causal effects of observed actions, while SPL is sensitive to the patterns of body motion involved in those actions. Additional analysis and explanation would help to determine the strength of evidence and the mechanistic underpinnings would benefit from closer consideration. Nevertheless, the work will be of broad interest to cognitive neuroscientists, particularly vision and action researchers.

eLife assessment

The authors have reported an important study in which they use a double-blind design to explore pharmacological manipulations in the context of a behavioral task. Despite a relatively small sample size, the findings are solid and motivate future explanations of the mechanism underlying their observations. The findings could be further strengthened by addressing some remaining concerns that relate to preprocessing, statistical details, and possible ocular artifacts.

eLife assessment

This study identified an innovative molecular mechanism linking diabetes to Alzheimer's disease (AD) risk, with important significance. The finding presents novel insights into AD pathogenesis and provides strong evidence about the mechanistic roles of Kallistatin, and the therapeutic potential of fenofibrate in AD. The experiments are well conducted, and the evidence is convincing.

eLife assessment

This valuable study investigates the selectivity of neuronal responses in the primary visual cortex and the dorsal lateral geniculate nucleus to stimuli presented far outside their receptive fields. The evidence supporting the claims is incomplete, due to lack of clarity. This paper should be of interest to neurophysiologists interested in vision and contextual modulations.

eLife assessment

This work presents important findings regarding the interaction of the monkeypox virus (MPXV) attachment H3 protein with the cellular receptor heparan sulfate and the use of this information to develop antivirals potentially effective against all orthopoxviruses. Using a combination of state-of-the art computational and wet experiments the authors present solid evidence to sustain their claims. These results will interest those working on basic orthopoxviruses biology and antiviral development.

eLife assessment

This study presents valuable framework and findings to our understanding of the brain cortex as a fractal object. Based on detailed methodology, the evidence provided on the stability of its shape property within 11 primate species is convincing, as well as the scale-specific effects of ageing on the human brain. This study will be of interest to neuroscientists interested in brain morphology, and to physicists and mathematicians interested in modeling the shapes of complex objects.

eLife assessment

This study presents a valuable report on a machine-learning tool for predicting synergistic drug combinations for cancer treatment. However, the evidence supporting the claims of the authors is incomplete, as the reported model shows some evidence of overfitting, and the claims of the authors could be strengthened if additional validation experiments were performed. The work will be of interest to oncologists and medical biologists working on cancer.

eLife assessment

This study reports a comparative biochemical and structural analysis of two PLP decarboxylase enzymes from plants. The work is useful because of the potential application of these enzymes in industrial theanine production. The structure provides a solid basis for understanding substrate specificity but some aspects of the work are incomplete. The paper will be of interest to enzymologists studying PLP enzymes and those working on enzyme engineering in plants.

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

Pharmacological induction of physiological slowing combined with organ perfusion systems could provide a novel therapeutic strategy for tissue and organ preservation. Using a Xenopus model, the authors provide important findings on a use of drug to slow down metabolism for the purpose of organ preservation. The authors provide compelling evidence that SNC80 can rapidly and reversibly slow biochemical and metabolic activities while preserving cell and tissue viability. This approach may be beneficial for transplantation, trauma management, and improving organ survival in remote and low-resource settings

eLife assessment

This important study highlights the importance of SLAM-SAP signaling in determining innate gamma-delta T cell sublineages and their T cell receptor repertoires. It uncovers the complex role of the SLAM-SAP pathway in developing specific gamma-delta T cell subsets. The evidence presented is compelling, backed by high-quality data obtained through advanced single cell proteogenomics techniques.This work will be of broad interest to immunologists.

eLife assessment

This valuable study reports that while most plasmacytoid dendritic cells (pDCs) originate from common dendritic cell precursors, approximately 20% are derived from lymphoid progenitors shared with B cells. The methodology used and the evidence are solid, and further demonstrate the distinct transcription factor requirements and activities of this subset of pDCs, although the functional significance of this dendritic cell subset will require further elucidation. The findings will be of great interest for those interested in the developmental and functional biology of the immune system.

eLife assessment

This important study examines E. coli growth and division, suggesting that inhomogeneous organization of ribosomes in the cytoplasm results in cell size-dependent growth rate perturbations. The work is conceptually appealing, but incomplete due to shortcomings in the experiments and modeling.

eLife assessment

Giamundo et al. present valuable data with new insights new insight into the role of Ezrin, a major membrane-actin linker that assembles signaling complexes, in the spatial regulation of EGF signaling mediators. The use of multiple state-of-the-art microscopy techniques, multiple cell lines and inhibitors, and in vivo models provides solid support for the majority of their conclusions. The findings are helpful for our understanding of EGF/mTOR signal transduction and support a critical role for the scaffolding protein Ezrin, in the upstream regulation of EGFR/AKT activity, TSC subcellular localization and mTORC1 signaling, thus contributing to our understanding of the regulation of endo-lysosomal signaling, alterations in which are implicated in many human diseases.

eLife assessment

This important work shows compelling data that significantly advances our understanding of the regulation of neurotransmitter and hormone secretion by exploring the mechanisms of how the protein complexin 2 (Cplx2) interacts with the calcium sensor synaptotagmin. The function of mammalian Cplx2 is studied using chromaffin cells derived from Cplx2 knock out mice as a system to overexpress and functionally characterize mutant Cplx2 forms and the interaction between Cplx2 and synaptotagmin. The authors identify structural requirements within the protein for Cplx's dual role in preventing premature vesicle exocytosis and enhancing evoked exocytosis. The findings are of broad interest to neuroscientists and cell biologists.

eLife assessment

Shore et al. report important effects of a heterozygous mutation in the KCNT1 potassium channel on ion currents and firing behavior of excitatory and inhibitory neurons in the cortex of KCNT1-Y777H mice. The authors provide solid evidence of physiological differences between this heterozygous mutation and their previous work with homozygotes. The reviewers appreciated the inclusion of recordings in ex vivo slices and dissociated cortical neurons, as well as the additional evidence showing an increase in persistent sodium currents (INaP) in parvalbumin-positive interneurons in heterozygotes. However, they were unclear regarding the likelihood of the increased sodium influx through INaP channels increasing sodium-activated potassium currents in these neurons.

eLife assessment

This important work proposes a neural network model of interactions between the prefrontal cortex and basal ganglia to implement adaptive resource allocation in working memory, where the gating strategies for storage are adjusted by reinforcement learning. Numerical simulations provide convincing evidence for the superiority of the model in improving effective capacity, optimizing resource management, and reducing error rates, as well as solid evidence for its human-like performance. The paper could be strengthened further by a more thorough comparison of model predictions with human behavior and by improved clarity in presentation. This work will be of broad interest to computational and cognitive neuroscientists, and may also interest machine-learning researchers who seek to develop brain-inspired machine-learning algorithms for memory.

eLife assessment

Rachubinski and colleagues provide an important manuscript that includes two major advances in understanding immune dysregulation in a large cohort of individuals with Down syndrome. The work comprises compelling, comprehensive, and state-of-the-art clinical, immunological, and autoantibody assessment of autoimmune/inflammatory manifestations. Additionally, the authors report promising results from a clinical trial with the JAK inhibitor tofacitinib for individuals with dermatological autoimmune disease.

eLife assessment

Based on analyses of retinae from genetically modified mice, and from wild-type ground squirrel and macaque, employing microscopic imaging, electrophysiology, and pharmacological manipulations, this valuable study on the role of Cav1.4 calcium channels in cone photoreceptor cells (i) shows that the expression of a Cav1.4 variant lacking calcium conductivity supports the development of cone synapses beyond what is observed in the complete absence of Cav1.4, and (ii) indicates that the cone pathway can partially operate even without calcium flux through Cav1.4 channels, thus preserving behavioral responses under bright light. The evidence for the function of Cav1.4 protein in synapse development is convincing and in agreement with a closely related earlier study by the same authors on rod photoreceptors. The mechanism of compensation of Cav1.4 loss by Cav3 remains unclear but appears to involve post-transcriptional processes. As congenital Cav1.4 dysfunction can cause stationary night blindness, this work relates to a wide range of neuroscience topics, from synapse biology to neuro-ophthalmology.

eLife assessment

The authors have presented an interesting set of results showing that female sex peptide signaling adversely affects late-life neurodegeneration after early-life exposure to repetitive mild head injury in Drosophila. This fundamental work substantially advances our understanding of how sex-dependent response to TBI occurs by identifying the Sex Peptide and the immune system as modulators of sex differences. The evidence supporting the conclusions is compelling with rigorous inclusion of controls and appropriate statistics.

eLife assessment

These valuable findings develop a mouse model with trackable fusion Fshr protein, which will be of use to the field. The animal model helps to elucidate the expression and function of the FSH receptor in extra-gonadal tissues. The strength of the evidence is solid in most parts, although additional validation of the localization data would strengthen the study considerably.

eLife assessment

This important study identifies a novel gastrointestinal enhancer of Ctnnb1. The authors present convincing evidence to support their claim that the dosage of Wnt/β-catenin signaling controlled by this enhancer is critical to intestinal epithelia homeostasis and the progression of colorectal cancers. The study will be of interest to biomedical researchers interested in Wnt signaling, tissue-specific enhancers, intestinal homeostasis, and colon cancer.

eLife assessment

The valuable work by authors improves our understanding on the effects of tree diversity on host-parasitoid communities of insects in forests in an experimental setting. Most of the analyses used are solid, but some of the conclusions seem a bit too strong and stretched.

eLife assessment

The manuscript presents a machine-learning method to predict protein hotspot residues. The validation is incomplete, along with the misinterpretation of the results with other current methods like FTMap.

eLife assessment

This useful study examines the neural activity in the motor cortex as a monkey reaches to intercept moving targets, focusing on how tuned single neurons contribute to an interesting overall population geometry. The presented results and analyses are solid, though the investigation of this novel task could be strengthened by clarifying the assumptions behind the single neuron analyses, and further analyses of the neural population activity and its relation to different features of behaviour.

eLife assessment

Semenova et al. have studied a large cross-sectional cohort of people living with HIV on suppressive antiretroviral therapy and performed high dimensional flow-cytometry for analysis with data science/machine learning approaches to investigate associations of immunological and clinical parameters and intact/total HIV DNA levels (and categorizations). The study is useful in introducing these new methods and large data set and appears mostly solid, though some of the claims were incompletely supported by the modeling results. The authors have revised the text to fairly reflect their results, yet open questions remain about utility, particularly as to the value of categorical classification (vs continuous measurement) of reservoir size.

eLife assessment

The present work provides new insights into detailed brain morphology. Using state-of-the-art methods, it provides compelling evidence for the relevance of sucal morphology for the precise localization of brain function. The fundamental findings have great relevance for the fields of imaging neuroscience and individualized medicine as ever-improving techniques improve precision to the point where individual brain anatomy is taking centre stage.

eLife assessment

This is an important study that connects the polymerase-associated factor 1 complex (Paf1C) with Histone 2B monoubiquitination and the expression of genes key to virulence in Cryptococcus neoformans. The provided information is convincing and has the potential to open several opportunities to further understand the basic biology of this significant human fungal pathogen.

eLife assessment

This study presents an important computational tool for the quantification of the cellular composition of human tissues profiled with ATAC-seq. The methodology is solid and its application results on breast cancer tumor tissues are convincing. It advances existing methods by utilizing a comprehensive reference profile for major cancer-relevant cell types, compatible with a widely-used cell type deconvolution tool.

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.

eLife assessment

This important study combines experiment and theory to examine how the intrinsic physiological properties of neurons involved in orchestrating birdsong are related to the temporal structure of song. Intrinsic properties determine how neurons respond to inputs, and in this manuscript, the authors describe rules that connect these intrinsic properties to a learned behaviour, the learned song of an adult songbird. The experimental data are convincing and the computational model builds on a robust and well-validated biophysical framework. Although some key points of the model could be established more strongly, 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

This preprint explores the involvement of cyclic di-GMP in genome stability and antibiotic persistence regulation in bacterial biofilms. The authors proposed a novel mechanism that, due to bacterial adhesion, increases c-di-GMP levels and influences persister formation through interaction with HipH. While the work may provide useful insights that could attract researchers in biofilm studies and persistence mechanisms, the main findings are inadequately supported and require further validation and refinement in experimental design.

eLife assessment

This study retrospectively analyzed clinical data to develop a risk prediction model for pulmonary hypertension in high-altitude populations. The evidence is solid and the findings are useful and hold clinical significance as the model can be used for intuitive and individualized prediction of pulmonary hypertension risk in these populations.

eLife assessment

This manuscript provides valuable evidence comparing the performance of mathematical models and opinions from experts engaged in outbreak response in forecasting the spatial spread of an Ebola epidemic. The evidence supporting the conclusions is convincing though the work might have benefited from the use of more than two models in the ensemble predictions. It will be of interest to disease modellers, infectious disease epidemiologists, policy-makers, and those who need to inform policy-makers during an outbreak.

eLife assessment

The authors report that chemogenetic methods targeting the ventral cervical spinal cord can be used to increase phrenic inspiratory motor output and subsequent diaphragm EMG activity and ventilation in rodents. These findings are important because they are a necessary first step towards using chemogenetic methods to drive inspiratory activity in disorders in which motor neurons are compromised, such as spinal injury and degenerative disease. The data are convincing, with rigorous assessments of phrenic inspiratory activity and its ability to drive the diaphragm and subsequent ventilation, as well as assessments of DREADD expression.

eLife assessment

This valuable study by Cui et al. investigates mechanisms generating sighs, which are crucial for respiratory function and linked to emotional states. Utilizing advanced methods in mice, they provide solid evidence that increased excitability in specific preBötzinger complex neuronal subpopulations expressing Neuromedin B receptors, gastrin-releasing peptide receptors, or somatostatin, can induce sigh-like large-amplitude inspirations. With additional technical clarifications and further supporting evidence for the implied capability of the neuron subpopulations studied to intrinsically generate the normal slow sigh rhythm, the study will interest neuroscientists studying respiratory neurobiology and rhythmic motor systems.

eLife assessment

This study reports a fundamental observation concerning cell death regulation by the anti-apoptotic BCL2 family NOXA. The authors convincingly demonstrate that NOXA is destabilized through the interaction with WSB2, a substrate receptor in CRL5 ubiquitin ligase complex, sensitizing the cells to treatments. These are key findings for cell biologists and cancer researchers as they identified a new target impacting drug responsiveness in cancer therapies.

eLife assessment

This study reports a valuable finding for the treatment of colorectal cancer (CRC), as the authors demonstrated that the enzyme CPT1A plays a significant role in the response to radiotherapy in CRC patients. The methodology and results presented by the authors are solid, supporting the role of CPT1A in CRC radiosensitivity, as the authors determined the expression of CPT1A in CRC tumors and non-tumor tissue, and they validated these findings with in vitro experiments.

eLife assessment

The authors provide useful data to support the existence of a regulatory pathway starting with SPI1-driven ZFP36L1 expression, that goes on to downregulate HDAC3 expression at the transcript level, leading to PD-L1 upregulation due to implied enhanced acetylation of its promoter region. This is therefore an interesting pathway that adds to our understanding of how PD-L1 expression is controlled in gastric cancer. However, this is likely one of many possible pathways that impact PD-L1 expression, and the data are currently incomplete to support the claims made.

eLife assessment

Early-life adversity or stress can enhance stress susceptibility by causing changes in emotion, cognition, and reward-seeking behaviors. This important manuscript highlights the involvement of lateral amygdala astrocytes in fear generalization and the associated synaptic plasticity, which are parallel to the effects of early life stress. With an elegant combination of behavioral models, morphological and functional assessments using immunostaining, electrophysiology, and viral-mediated loss-of-function approaches, the authors provide solid correlational and causal evidence that is consistent with the hypothesis that early life stress produces neural and behavioral dysfunction via perturbing lateral amygdala astrocytic function.

eLife assessment

This manuscript provides convincing evidence derived from diverse state-of-the-art approaches to suggest that non-dopaminergic projection neurons in the ventral tegmental area (VTA) make local synapses. These important findings challenge the prevailing wisdom that VTA interneurons exclusively form local synaptic contacts and instead reveal that VTA neurons expressing interneuron markers also form long-range projections to forebrain targets such as the cortex, ventral pallidum, and nucleus accumbens. Given the importance of VTA interneurons to many models of VTA-linked behavioral functions, these findings have significant implications for our understanding of the neural circuits underlying reward, motivation, and addiction.

eLife assessment

Franke et al. explore and characterize color response properties of neurons in mouse primary visual cortex (V1), revealing specific color opponent encoding strategies across the visual field. The paper provides evidence for the existence of color opponency in a subset of neurons within V1 and shows that these color opponent neurons are more numerous in the upper visual field. Support for the main conclusions is convincing and the dataset that forms the basis of the paper is impressive. The paper will make an important contribution to understanding how color is coded in mouse V1.

eLife assessment

This valuable work uses unbiased approaches to discover critical molecules in C. elegans and its bacterial food for nutrition sensing and food choice, providing a framework for other studies. The data convincingly support their model that C. elegans uses UPRER and immune response pathways to evaluate sugar contents in the bacteria to change their behaviors.

eLife assessment

The findings presented by the authors are useful within the focused scope of endometriosis treatment, providing a potential new therapeutic approach. The strength of the evidence is, however, incomplete, as the main claims are only partially supported by the authors' data. The research nevertheless offers promising initial evidence for KMO inhibition as a novel non-hormonal therapy for endometriosis, but further studies are needed to confirm efficacy and address any potential side effects.