An optimized workflow for transcriptomic analysis from archival paraformaldehyde-fixed retinal tissues collected by laser capture microdissection.

RNA sequencing (RNA-seq) coupled with laser capture microdissection (LCM) is a powerful tool for transcriptomic analysis in unfixed fresh-frozen tissues. Fixation of ocular tissues for immunohistochemistry commonly involves the use of paraformaldehyde (PFA) followed by embedding in Optimal Cutting Temperature (OCT) medium for long-term cryopreservation. However, the quality of RNA derived from such archival PFA-fixed/OCT-embedded samples is often compromised, limiting its suitability for transcriptomic studies. In this study, we aimed to develop a methodology to extract high-quality RNA from PFA-fixed canine eyes by utilizing LCM to isolate retinal tissue. We demonstrate the efficacy of an optimized LCM and RNA purification protocol for transcriptomic profiling of PFA-fixed retinal specimens. We compared four pairs of canine retinal tissues, where one eye was subjected to PFA-fixation prior to OCT embedding, while the contralateral eye was embedded fresh frozen (FF) in OCT without fixation. Since the RNA obtained from PFA-fixed retinas were contaminated with genomic DNA, we employed two rounds of DNase I treatment to obtain RNA suitable for RNA-seq. Notably, the quality of sequencing reads and gene sets identified from both PFA-fixed and FF tissues were nearly identical. In summary, our study introduces an optimized workflow for transcriptomic profiling from PFA-fixed archival retina. This refined methodology paves the way for improved transcriptomic analysis of preserved ocular tissue, bridging the gap between optimal sample preservation and high-quality RNA data acquisition.

Decision making for health-related research outcomes that alter diagnosis: A model from paediatric brain tumours.

AIMS: The question of how to handle clinically actionable outcomes from retrospective research studies is poorly explored. In neuropathology, this problem is exacerbated by ongoing refinement in tumour classification. We sought to establish a disclosure threshold for potential revised diagnoses as determined by the neuro-oncology speciality. METHODS: As part of a previous research study, the diagnoses of 73 archival paediatric brain tumour samples were reclassified according to the WHO 2016 guidelines. To determine the disclosure threshold and clinical actionability of pathology-related findings, we conducted a result-evaluation approach within the ethical framework of BRAIN UK using a surrogate clinical multidisciplinary team (MDT) of neuro-oncology specialists. RESULTS: The MDT identified key determinants impacting decision-making, including anticipated changes to patient management, time elapsed since initial diagnosis, likelihood of the patient being alive and absence of additional samples since cohort inception. Ultimately, none of our research findings were considered clinically actionable, largely due to the cohort's historic archival and high-risk nature. From this experience, we developed a decision-making framework to determine if research findings indicating a change in diagnosis require reporting to the relevant clinical teams. CONCLUSIONS: Ethical issues relating to the use of archival tissue for research and the potential to identify actionable findings must be carefully considered. We have established a structured framework to assess the actionability of research data relating to patient diagnosis. While our specific findings are most applicable to the pathology of poor prognostic brain tumour groups in children, the model can be adapted to a range of disease settings, for example, other diseases where research is dependent on retrospective tissue cohorts, and research findings may have implications for patients and families, such as other tumour types, epilepsy-related pathology, genetic disorders and degenerative diseases.

Publishing publicly available interview data: an empirical example of the experience of publishing interview data.

In September 2021 I made a collection of interview transcripts available for public use under a CreativeCommons license through the Princeton DataSpace. The interviews include 39 conversations I had with gig workers at AmazonFlex, Uber, and Lyft in 2019 as part of a study on automation efforts within these organizations. I made this decision because (1) I was required to contribute to a publicly available data set as a requirement of my funding and (2) I saw it as an opportunity to engage in the collaborative qualitative science experiments emerging in Science and Technology studies. This article documents my thought process and step-by-step design decisions for designing a study, gathering data, masking it, and publishing it in a public archive. Importantly, once I decided to publish these data, I determined that each choice about how the study would be designed and implemented had to be assessed for risk to the interviewee in a very deliberate way. It is not meant to be comprehensive and cover every possible condition a researcher may face while producing qualitative data. I aimed to be transparent both in my interview data and the process it took to gather and publish these data. I use this article to illustrate my thought process as I made each design decision for this study in hopes that it could be useful to a future researcher considering their own data publishing process.

Long-distance migrations and seasonal movements of meagre (Argyrosomus regius), a large coastal predator, along the Iberian Peninsula coast.

BACKGROUND: The meagre, Argyrosomus regius, is a large coastal predatory fish inhabiting waters from the north-eastern Atlantic and Mediterranean Sea, where it is targeted by commercial and recreational fisheries. Previous genetic studies have found an unexpectedly high population differentiation not only between the Atlantic and the Mediterranean, but also along the Atlantic coast. However, the reasons underpinning this genetic barrier remained unclear. Likewise, even though the species is amongst the world's largest marine teleosts, knowledge about its movement ecology and migratory behaviour remains notably scarce, and primarily reliant on fisheries-dependent data. METHODS: In this study, we used a combination of acoustic telemetry and pop-up satellite archival tags to investigate the movements of 22 adult meagre (70-143 cm total length) along the Southwestern coast of the Iberian Peninsula. RESULTS: Our results strongly suggest that the previously reported genetic differentiation is not maintained by limited adult dispersal/movement, as hypothesized. On the contrary, we documented some of the longest individual annual migrations ever recorded for a coastal teleost, up to > 2000 km, with frequent back-and-forth movements between the West and Southern Iberian coasts. Moreover, their detected regional movement patterns support the existence of a marked seasonal behavioural shift, with individuals being less active and moving to deeper waters during winter, and are consistent with spawning philopatry associated to their summer reproductive movements. Finally, we identified putative aggregation areas that may harbour important feeding/overwintering grounds. CONCLUSIONS: These findings shed new light on the movement and behaviour patterns of meagre that may be of particular importance for the conservation and spatial management of this species throughout its range, and open the door to further research on functional connectivity.

NKX2-2 based nuclei sorting on frozen human archival pancreas enables the enrichment of islet endocrine populations for single-nucleus RNA sequencing.

BACKGROUND: Current approaches to profile the single-cell transcriptomics of human pancreatic endocrine cells almost exclusively rely on freshly isolated islets. However, human islets are limited in availability. Furthermore, the extensive processing steps during islet isolation and subsequent single cell dissolution might alter gene expressions. In this work, we report the development of a single-nucleus RNA sequencing (snRNA-seq) approach with targeted islet cell enrichment for endocrine-population focused transcriptomic profiling using frozen archival pancreatic tissues without islet isolation. RESULTS: We cross-compared five nuclei isolation protocols and selected the citric acid method as the best strategy to isolate nuclei with high RNA integrity and low cytoplasmic contamination from frozen archival human pancreata. We innovated fluorescence-activated nuclei sorting based on the positive signal of NKX2-2 antibody to enrich nuclei of the endocrine population from the entire nuclei pool of the pancreas. Our sample preparation procedure generated high-quality single-nucleus gene-expression libraries while preserving the endocrine population diversity. In comparison with single-cell RNA sequencing (scRNA-seq) library generated with live cells from freshly isolated human islets, the snRNA-seq library displayed comparable endocrine cellular composition and cell type signature gene expression. However, between these two types of libraries, differential enrichments of transcripts belonging to different functional classes could be observed. CONCLUSIONS: Our work fills a technological gap and helps to unleash frozen archival pancreatic tissues for molecular profiling targeting the endocrine population. This study opens doors to retrospective mappings of endocrine cell dynamics in pancreatic tissues of complex histopathology. We expect that our protocol is applicable to enrich nuclei for transcriptomics studies from various populations in different types of frozen archival tissues.

Effect of formalin-fixation and paraffin-embedded tissue storage times on RNAscope in situ hybridization signal amplification.

RNAscope in situ hybridization (ISH) detects target RNA in formalin-fixed, paraffin-embedded (FFPE) tissues. Protocols suggest that prolonged FFPE storage and formalin fixation may impact signal detection, potentially limiting the utility of RNAscope ISH in retrospective studies. To develop parameters for RNAscope use with archived specimens, we evaluated the effect of formalin-fixation time by measuring the signal of a reference gene (16srRNA) in selected tissues fixed in 10% neutral-buffered formalin for 1, 2, 3, 5, 7, 10, 14, 21, 28, 60, 90, 180, and 270 d. The signal intensity and percent area of signal decreased after 180 d. Tissues had detectable signal at 180 d but not at 270 d of formalin fixation. To assess target detection in paraffin blocks, we qualitatively compared the signal of canine distemper virus (CDV) antigen via immunohistochemistry and CDV RNA via RNAscope ISH in replicate sections from blocks stored at room temperature for 6 mo, 1, 3, 6, 8, 11, 13, and 15 y; RNA was detected in FFPE tissues stored for up to 15 y. Our results demonstrate that RNAscope ISH can detect targets in tissues with prolonged paraffin storage intervals and formalin-fixation times.

An Historical Case Study of Trans Exclusion and Empowerment: Implications for Transgender Older Adults and Aging.

Transgender older adults have a long history of exclusion that shapes current experiences with social services. However, scant gerontological research uses archival data, which can provide critical context for service providers. Moreover, sparse research examines how exclusion can be a catalyst for change that social workers could leverage. Empowerment theory provides a theoretical tool to explain how this is possible. This multidisciplinary case study blends community member interviews and archival data to answer this question: How did exclusion shape empowerment and social change for transgender Americans? This study focuses on the events before and after the Compton's Cafeteria Riot, one of the first acts of LGBTQIA+ collective resistance that led to new services for and by transgender Americans. Data reveal how exclusion facilitated the emergence of collective empowerment among transgender women and queer youth in San Francisco. Archival data shows how exclusion preceded self-efficacy, critical consciousness, involvement with similar others, acquisition of new skills, and ultimately action to eliminate social, economic, and political barriers and power imbalances. This study provides both empirical and theoretical tools to contribute new data and perspectives on trans exclusion and empowerment and its implications for social workers serving transgender older adults.

Exploring general practice research in Germany: a systematic review of dissertation topics from 1965-2023.

BACKGROUND: Since its academic inception in the 1960s, Germany's general practice has seen numerous dissertations, many of which are housed in the 'Archive of German language General Practice' (ADAM). AIM: This study aims to provide the first comprehensive overview of dissertation topics from the discipline of general- and family medicine in Germany, establishing a foundation for advancing research. METHOD: We employed a systematic review approach, examining 801 dissertations from both ADAM and online sources. Each topic was identified, categorized, and finalized through consensus by two independent reviewers. RESULTS: Our analysis encompassed 486 dissertations from ADAM, 176 from the German national library, and 139 from university libraries. A total of 167 unique research topics were identified. The predominant themes included medical education (n = 49), medication orders (n = 39), frequent consultation issues (n = 33), complementary medicine (n = 32), and screening measures (n = 29). The use of qualitative methods was constantly rising, from no qualitative methods used from 1965-1974, up to 22% of dissertations in recent years. CONCLUSION: The diversity of 167 research topics underscores the vastness and complexity of general practice in Germany. This structured overview is pivotal for facilitating focused and interconnected research endeavors in the field.

[Historical aspects of the formation and development of children's health resort medicine in the Samara region]. / Istoricheskie aspekty formirovaniya i razvitiya detskoi kurortnoi meditsiny v Samarskom regione.

Archive documents and sources in scientific electronic databases were analyzed in order to study the historical origins of the health resort business in terms of children's health resort medicine in the Samara government. It has been established that the period of origin and intensive development of balneology for children in the Samara region occurred at the beginning of the 20th century and the first children's sanatoriums began to function thanks to the initiative of the Society of Governmental Physicians for charitable funds of the Samara nobility and merchantry in territories with natural and climatic therapeutic factors, namely Sernovodsky resort, Barboshina glade and Postnikov ravine. A comparative analysis of the dynamics of morbidity structure of children admitted for sanatorium treatment, as well as general education institutions, from which information on the health of school pupils is received, has been conducted over a period of 120 years. Despite the change of years and generations, there is a general concept of sanatorium health improvement of school pupils as a future social and economic pillar of society, with the formation of a healthy generation for both the region and the country in general.

The Diversity of Parasitoids and Their Role in the Control of the Siberian Moth, Dendrolimus sibiricus (Lepidoptera: Lasiocampidae), a Major Coniferous Pest in Northern Asia.

The Siberian moth, Dendrolimus sibiricus Tschetv., 1908 (Lepidoptera: Lasiocampidae) is a conifer pest that causes unprecedented forest mortality in Northern Asia, leading to enormous ecological and economic losses. This is the first study summarizing data on the parasitoid diversity and parasitism of this pest over the last 118 years (1905-2022). Based on 860 specimens of freshly reared and archival parasitoids, 16 species from two orders (Hymenoptera and Diptera) were identified morphologically and/or with the use of DNA barcoding. For all of them, data on distribution and hosts and images of parasitoid adults are provided. Among them, the braconid species, Meteorus versicolor (Wesmael, 1835), was documented as a parasitoid of D. sibiricus for the first time. The eastern Palaearctic form, Aleiodes esenbeckii (Hartig, 1838) dendrolimi (Matsumura, 1926), status nov., was resurrected from synonymy as a valid subspecies, and a key for its differentiation from the western Palaearctic subspecies Aleiodes esenbeckii ssp. esenbecki is provided. DNA barcodes of 11 parasitoid species from Siberia, i.e., nine hymenopterans and two dipterans, represented novel records and can be used for accurate molecular genetic identification of species. An exhaustive checklist of parasitoids accounting for 93 species associated with D. sibirisus in northern Asia was compiled. Finally, the literature and original data on parasitism in D. sibiricus populations for the last 83 years (1940-2022) were analysed taking into account the pest population dynamics (i.e., growth, outbreak, decline, and depression phases). A gradual time-lagged increase in egg and pupal parasitism in D. sibiricus populations was detected, with a peak in the pest decline phase. According to long-term observations, the following species are able to cause significant mortality of D. sibiricus in Northern Asia: the hymenopteran egg parasitoids Telenomus tetratomus and Ooencyrtus pinicolus; the larval parasitoids Aleiodes esenbeckii sp. dendrolimi, Cotesia spp., and Glyptapanteles liparidis; and the dipteran pupal parasitoids Masicera sphingivora, Tachina sp., and Blepharipa sp. Their potential should be further explored in order to develop biocontrol programs for this important forest pest.

Wide-spread dispersal in a deep-sea brooding polychaete: the role of natural history collections in assessing the distribution in quill worms (Onuphidae, Annelida).

BACKGROUND: Modern integrative taxonomy-based annelid species descriptions are detailed combining morphological data and, since the last decades, also molecular information. Historic species descriptions are often comparatively brief lacking such detail. Adoptions of species names from western literature in the past led to the assumption of cosmopolitan ranges for many species, which, in many cases, were later found to include cryptic or pseudocryptic lineages with subtle morphological differences. Natural history collections and databases can aid in assessing the geographic ranges of species but depend on correct species identification. Obtaining DNA sequence information from wet-collection museum specimens of marine annelids is often impeded by the use of  formaldehyde and/or long-term storage in ethanol resulting in DNA degradation and cross-linking. RESULTS: The application of ancient DNA extraction methodology in combination with single-stranded DNA library preparation and target gene capture resulted in successful sequencing of a 110-year-old collection specimen of quill worms. Furthermore, a 40-year-old specimen of quill worms was successfully sequenced using a standard extraction protocol for modern samples, PCR and Sanger sequencing. Our study presents the first molecular analysis of Hyalinoecia species including the previously known species Hyalinoecia robusta, H. tubicloa, H. artifex, and H. longibranchiata, and a potentially undescribed species from equatorial western Africa morphologically indistinguishable from H. tubicola. The study also investigates the distribution of these five Hyalinoecia species. Reassessing the distribution of H. robusta reveals a geographical range covering both the Atlantic and the Indian Oceans as indicated by molecular data obtained from recent and historical specimens. CONCLUSION: Our results represent an example of a very wide geographical distribution of a brooding deep-sea annelid with a complex reproduction strategy and seemingly very limited dispersal capacity of its offspring, and highlights the importance of molecular information from museum specimens for integrative annelid taxonomy and biogeography.

Introduced and extinct: neglected archival specimens shed new light on the historical biogeography of an iconic avian species in the Mediterranean.

Collection specimens provide valuable and often overlooked biological material that enables addressing relevant, long-unanswered questions in conservation biology, historical biogeography, and other research fields. Here, we use preserved specimens to analyze the historical distribution of the black francolin (Francolinus francolinus, Phasianidae), a case that has recently aroused the interest of archeozoologists and evolutionary biologists. The black francolin currently ranges from the Eastern Mediterranean and the Middle East to the Indian subcontinent, but, at least since the Middle Ages, it also had a circum-Mediterranean distribution. The species could have persisted in Greece and the Maghreb until the 19th century, even though this possibility had been questioned due to the absence of museum specimens and scant literary evidence. Nevertheless, we identified four 200-year-old stuffed black francolins-presumably the only ones still existing-from these areas and sequenced their mitochondrial DNA control region. Based on the comparison with conspecifics (n = 396) spanning the entirety of the historic and current species range, we found that the new samples pertain to previously identified genetic groups from either the Near East or the Indian subcontinent. While disproving the former occurrence of an allegedly native westernmost subspecies, these results point toward the role of the Crown of Aragon in the circum-Mediterranean expansion of the black francolin, including the Maghreb and Greece. Genetic evidence hints at the long-distance transport of these birds along the Silk Road, probably to be traded in the commerce centers of the Eastern Mediterranean.

Archival records of the Antarctic clam shells from Marian Cove, King George Island suggest a protective mechanism against ocean acidification.

Continuous emissions of anthropogenic CO2 are changing the atmospheric and oceanic environment. Although some species may have compensatory mechanisms to acclimatize or adapt to the changing environment, most marine organisms are negatively influenced by climate change. In this study, we aimed to understand the compensatory mechanisms of the Antarctic clam, Laternula elliptica, to climate-related stressors by using archived shells from 1995 to 2018. Principal component analysis revealed that seawater pCO2 and salinity in the Antarctic Ocean, which have increased since the 2000's, are the most influential factors on the characteristics of the shell. The periostracum thickness ratio and nitrogen on the outermost surface have increased, and the dissolution area (%) has decreased. Furthermore, the calcium content and mechanical properties of the shells have not changed. The results suggest that L. elliptica retains the mechanism of protecting the shell from high pCO2 by thickening the periostracum as a phenotype plasticity.

Bohemia at the Pacific seabed: Archiving the future of deep-sea mining with the Interoceanmetal Joint Organization.

This article uses historical and ethnographic methods to examine the primarily East-Central European Interoceanmetal Joint Organization (IOM). I ask how and why the IOM has survived as an institution since its inception in 1987, working especially with the personal archive of Vratislav Kubista. Kubista was a metallurgist and former Deputy Director General at IOM who after retirement sought to develop a local deep-sea mining museum. This is a story about the work that archives do, but even more about how institutions maintain archives. I draw on recent work in the history and anthropology of time and archival practice to situate IOM's history and Kubista's collection in narratives of ruin, the unbuilt, and the experience of multiple temporalities within spaces of resource speculation and anticipation. I suggest that IOM's history highlights the contingencies of resources in the temporality of indefinite pause, their attendant data, and scientific labor and life under the shifting political, economic, and scientific circumstances of the ongoing not-yet. In broadening the history of what is once more a hotly contested potential resource, this account speaks to the claims of contemporary would-be seabed miners, who frequently frame the practice in terms of innovation, urgency, and novelty.

Emotional responses in archival work.

Building on previous work investigating the impact of exposure to (a) records with traumatic potentialities and (b) interactions with donors and community researchers whose suffering is documented in the archives, this study sought to better understand emotional aspects of archival work. Using a diary research methodology, 15 archivists engaged in diary keeping for approximately four months. What emerged was a broad set of events and experiences that triggered a wide range of emotional responses arising from archival work. This included: pre-existing emotional states and characterological traits; emotional exchanges in the workplace with colleagues and others; emotional demands of the work (including emotion work and emotional labour); team and leader interactions arising from group tasks and leader behaviour; and organizational policies, climate, resources and demands. This broader set of interactional factors forms the foundation on which traumatic and other troubling events are encountered. Future research must consider the nature of archival organizations and interactions within them that contribute to the overall working experience. In addition, archival organizations need to take responsibility for creating a culture that demonstrates respect and appreciation for workers, acknowledges the interpersonal challenges of the work, and provides supports for archivists who are shouldering the challenges.

Archival skin biopsy specimens as a tool for miRNA-based diagnosis: Technical and post-analytical considerations.

Archived specimens, taken by standardized procedures in clinical practice, represent a valuable resource in translational medicine. Their use in retrospective molecular-based studies could provide disease and therapy predictors. Microfluidic array is a user-friendly and cost-effective method allowing profiling of hundreds of microRNAs (miRNAs) from a low amount of RNA. However, even though tissue miRNAs may include potentially robust biomarkers, non-uniformed post-analytical pipelines could hinder translation into clinics. In this study, epidermal RNA from archival skin biopsy specimens was isolated from patients with peripheral neuropathy and healthy individuals. Unbiased miRNA profiling was performed using RT-qPCR-based microfluidic array. We demonstrated that RNA obtained from archival tissue is appropriate for miRNA profiling, providing evidence that different practices in threshold selection could significantly influence the final results. We showed the utility of software-based quality control for amplification curves. We revealed that selection of the most stable reference and the calculation of geometric mean are suitable when utilizing microfluidic arrays without known references. By applying appropriate post-analytical settings, we obtained miRNA profile of human epidermis associated with biological processes and a list of suitable references. Our results, which outline technical and post-analytical considerations, support the broad use of archived specimens for miRNA analysis to unravel disease-specific molecular signatures.

Country's value priorities in health crisis: How dominant societal motivations shape COVID-19 severity.

This paper presents two comprehensive studies examining how Schwartz's human values dimensions at the country level predict COVID-19 pandemic severity. Study 1 aggregated survey data across 89 countries from the European Social Survey and World Values Survey to assess societal-level conservation versus openness to change (CON-OTC) and self-enhancement versus self-transcendence (SE-ST) value-continuums. Study 2 developed an innovative archival measurement approach using 10 indicators to estimate these value dimensions for over 180 countries. Both studies employed multilevel modeling to test the relationships between country-level values and COVID-19 severity, measured through epidemiological indicators of transmission speed, case fatality rate, infection prevalence and mortality burden. Results revealed that the CON-OTC and SE-ST value-continuums showed consistent, significant negative associations with transmission speed and infection prevalence before adjusting for modernization, latitude, historical pathogen prevalence and government stringency across both studies. However, after accounting for these socioecological and policy covariates, the CON-OTC value-continuum positively predicted case fatality rate in both studies, implying conservation values could increase COVID-19 lethality. In contrast, across both studies, the SE-ST value-continuum negatively predicted case fatality rate after adjusting for the covariates, suggesting countries prioritizing self-enhancement values exhibited relatively lower pandemic severity and lethality when accounting for developmental, ecological, and policy factors. Accordingly, the studies advance theoretical understanding of how country's value priorities shape COVID-19 impact. Methodologically, these studies contribute through multilevel techniques that account for spatial dependencies, as well as an innovative ecological measurement. Overall, this research demonstrates the value of applying Schwartz's framework at a societal level to predict global health crises and pandemics.

Tidal and circadian patterns of European eel during their spawning migration in the North Sea and the English Channel.

Technological advances in tracking methods enable the mapping of anguillid eel migration routes from continental habitats to their spawning sites in the ocean. However, the behaviour and orientation abilities of anguillids are still poorly understood, and have only rarely been studied on the continental shelf. Here we present the results of a study into the vertical and horizontal movement behaviour of 42 European eels (Anguilla anguilla L.) tagged with electronic tags that migrated through the North Sea and English Channel towards and into the Atlantic Ocean during their spawning migration. We used actograms, periodograms and linear mixed effects models to determine the periodicity and significance of the timing and pattern of vertical movement and activity. Overall, eels had a complex behavioural repertoire that included classical diel vertical migration (DVM), reverse DVM and vertical movement behaviours that synchronized with tidal patterns. All of the eels that were tracked showed one or more of these behaviours during their time at liberty, and many exhibited all of them. We also observed that the eels had a higher horizontal migration speed when the current in the favourable direction was stronger. This, together with the vertical movement synchronized with the tides, suggests the eels adopt selective tidal stream transport. Finally, tracked eels had a higher vertical movement range at night compared to daytime. We hypothesize that these behaviours are driven by bio-energetic efficient movement, navigation and predator avoidance.

Pictures at an exhibition: How to share your imaging data.

Open access to data underpinning published results is a key pillar of scientific reproducibility. Making data available at scale also provides opportunities for data reuse, encouraging the development of new analysis approaches. In this poster article, accompanying a recorded talk, we will explain the benefits of publicly archiving your image data alongside your published manuscripts, as well as highlight what resources are available to do this. This will include the BioImage Archive, EMBL-EBI's new resource for biological image data, https://www.ebi.ac.uk/bioimage-archive/. We will look at how image data submission works, how to prepare in advance for archiving your data and upcoming developments.

Movement and vertical habitat use of yellowfin tuna Thunnus albacares in a vertically compressed habitat: the Galápagos Marine Reserve.

Tropical pelagic predators are exploited by fisheries and their movements are influenced by factors including prey availability, temperature, and dissolved oxygen levels. As the biophysical parameters vary greatly within the range of circumtropical species, local studies are needed to define those species' habitat preference and model possible behavioral responses under different climate change scenarios. Here, we tagged yellowfin tuna Thunnus albacares in the Galápagos Marine Reserve and tracked the horizontal and vertical movements of eight individuals for 4-97 days. The tuna traveled a mean of 13.6 km day-1 horizontally and dispersed throughout the archipelago and in offshore waters inside the Galápagos Marine Reserve and in the surrounding Ecuadorian exclusive economic zone. Vertically, they traveled a mean of 2 km day-1 , although high-resolution data from a recovered tag suggested that transmitted data underestimated their vertical movement by a factor of 5.5. The tracked yellowfin tuna spent most of their time near the surface, with an overall mean swimming depth of 24.3 ± 46.6 m, and stayed shallower at night (11.1 ± 16.3 m) than during the day ( 37.7 ± 60.9 m), but on occasion dived to cold, oxygen-poor waters below 200 m. Deep dives were commonly made during the day with a mean recovery period of 51 min between exposures to modeled oxygen-limiting conditions <1.5 mL L-1 , presumably to re-oxygenate. The depth and frequency of dives were likely limited by dissolved oxygen levels, as oxygen-depleted conditions reach shallow depths in this region. The main habitat of tracked yellowfin tunas was in the shallow mixed layer, which may leave them vulnerable to fishing. Vertical expansion of low-oxygen waters under future climate change scenarios may further compress their habitat, increasing their vulnerability to surface fishing gear.