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Few animal groups can claim the level of wonder that cephalopods instill in the minds of researchers and the general public. Much of cephalopod biology, however, remains unexplored: the largest invertebrate brain, difficult husbandry conditions, and complex (meta-)genomes, among many other things, have hindered progress in addressing key questions. However, recent technological advancements in sequencing, imaging, and genetic manipulation have opened new avenues for exploring the biology of these extraordinary animals. The cephalopod molecular biology community is thus experiencing a large influx of researchers, emerging from different fields, accelerating the pace of research in this clade. In the first post-pandemic event at the Cephalopod International Advisory Council (CIAC) conference in April 2022, over 40 participants from all over the world met and discussed key challenges and perspectives for current cephalopod molecular biology and evolution. Our particular focus was on the fields of comparative and regulatory genomics, gene manipulation, single-cell transcriptomics, metagenomics, and microbial interactions. This article is a result of this joint effort, summarizing the latest insights from these emerging fields, their bottlenecks, and potential solutions. The article highlights the interdisciplinary nature of the cephalopod-omics community and provides an emphasis on continuous consolidation of efforts and collaboration in this rapidly evolving field.

Summary: Background: A better understanding of the molecular events during liver normothermic machine perfusion (NMP) is warranted to develop a data-based approach for the identification of biomarkers representative of graft quality and posttransplant outcome. We analysed the dynamic transcriptional changes during NMP and linked them to clinical and biochemical parameters. Methods: 50 livers subjected to NMP for up to 24 h were enrolled. Bulk RNA sequencing was performed in serial biopsies collected pre and during NMP, and after reperfusion. Perfusate was sampled to monitor liver function. qPCR and immunohistochemistry were performed to validate findings. Molecular profiles were compared between transplanted and non-transplanted livers, and livers with and without early allograft dysfunction. Findings: Pathways related to immune and cell stress responses, cell trafficking and cell regulation were activated during NMP, while cellular metabolism was downregulated over time. Anti-inflammatory responses and genes involved in tissue remodelling were induced at later time-points, suggesting a counter-response to the immediate damage. NMP strongly induced a gene signature associated with ischemia-reperfusion injury. A 7-gene signature corresponds with the benchmarking criteria for transplantation or discard at 6 h NMP (area under curve 0.99). CD274 gene expression (encoding programmed cell-death ligand-1) showed the highest predictive value. LEAP2 gene expression at 6 h NMP correlated with impaired graft function. Interpretation: Assessment of gene expression markers could serve as a reliable tool to evaluate liver quality during NMP and predicts early graft function after transplantation. Funding: The research was supported by “In Memoriam Dr. Gabriel Salzner Stiftung”, Tiroler Wissenschaftsfond, Jubiläumsfonds-Österreichische Nationalbank and MUI Start grant.

Neoplastic processes of the mandible and their treatment are rarely reported in large animal species. Specifically, giant cell tumor of bone is an uncommon tumor in animals and has been associated in humans with locally invasive behavior and a high recurrence rate. En-bloc resection is the treatment of choice, but depending on the localization of the tumor, this may result in functional deficits. This report details the diagnostic work-up, treatment, and long-term outcome of a giant cell tumor of bone involving the rostral mandible and mandibular symphysis of a goat. Extensive rostral mandibulectomy involving the entire mandibular symphysis without surgical fixation of the hemimandibles was performed. Histological and electron microscopic findings of the tumor were consistent with a giant cell tumor of bone. Although a mutation of the H3F3A gene is considered the driver of tumor development in human giant cell tumors, using molecular analysis, this gene mutation could not be confirmed in this case. Follow-up examinations revealed spontaneous secondary fusion of both hemimandibles and no signs of tumor recurrence. Nearly 1 year after surgery, the owners reported no signs of tumor regrowth.

The European Union of Aquarium Curators (EUAC) boasts a membership of 150 dedicated individuals, standing as a vital cornerstone within the European public aquarium community. Since 2004, the EUAC Conservation Fund has granted over a quarter of a million euros to approximately 50 conservation projects spanning the globe. These initiatives, diverse in nature and scale, have yielded tangible impacts on local populations and their focal species. This paper delves into the outcomes of these conservation endeavors and proposes enhancements to ensure that the funding is unequivocally channeled towards conservation efforts. One resounding observation gleaned from the array of projects spotlighted in this study is the profound community engagement that emerges, irrespective of the final project outcomes. These endeavors serve as a catalyst for local communities, shedding light on subjects that would otherwise remain shrouded in obscurity. Furthermore, the EUAC-backed projects illuminate the expansive reach of public aquarium initiatives, transcending the confines of acrylic tank walls and institutional boundaries to resonate globally, heightening local awareness about the imperative to safeguard biodiversity. These findings underscore a prospective trajectory for both the EUAC and the public aquaria it comprises: an intensified advocacy and collaboration with legislative bodies to fortify in situ conservation measures. In essence, it is imperative that the public comprehends the pivotal role played by aquaria in preserving a multitude of species and acknowledges that their visits directly contribute to funding projects aimed at safeguarding species within their natural habitats.

Morphological differences between males and females are common among reptiles. A particularly interesting sexually dimorphic feature whose function is largely unknown is the number and pattern of specific scales. Several lizard species possess an arrangement of centered scales near the cloacal region that differ between the sexes and can be used for sex determination. The presence of postcloacal buttons, sexually dimorphic postcloacal scales on both sides of the body, is an exclusive trait in the subfamily Tupinambinae and is only poorly documented. Here, we investigate postcloacal scales in northern caiman lizards (Dracaena guianensis) housed at the Vienna Zoo. For a period of two years, we documented scale patterns and performed morphometric measurements of individuals of different age classes. Caiman lizards were CT scanned to confirm the sexes. Males exhibit three raised postcloacal scales in a row behind the left and right leg, while females possess one or two large scales surrounded by several smaller scales. The study provides the first evidence that these scales can function as a reliable trait to distinguish the sexes regardless of age or reproductive status. The sexually dimorphic bilateral scale pattern is present immediately after hatching and does not change during development. Scales only increase in thickness and length during growth. We further demonstrate that sexual size dimorphism (SSD) exists in juveniles during ontogenetic development. Juvenile females had a larger SVL, body length, tail length and higher weight compared to juvenile males. This SSD could not be confirmed in adults, and sex determination based on SSD seems unreliable.

Adequate adaptions and actions to combat anthropogenic climate change (CC) are significant challenges of the 21st century. In Europe, according to the European Environmental Agency, warming of around 2°C is expected under the moderate climate scenario (RCP 4.5) by the end of the century, but the pessimistic RCP 8.5 scenario project an increase of up to 6°C. In addition to the rise in temperature, changes in precipitation and increased frequency of extreme weather events are predicted. New environmental conditions affect tree species and habitats differently; thus, forest biodiversity and local tree species compositions probably will be altered in many regions in the future. The effects may be manifold: some tree species may persist, locally adapt and migrate, while others may disappear from given regions and be replaced by native or non-native species. The native forests of the Austrian-Hungarian border region are particularly affected by the climate change. To mitigate the consequences of anthropogenic climate change to preserve forest biodiversity for future generations and to enable their use, deliberate and planned human interventions and actions are essential. These require transnational or even global efforts since nature and climate do not recognise man-made borders.The REIN-Forest project (Interreg V-A Austria-Hungary Programme - ATHU150), a bilateral project between Austria and Hungary, aimed to establish harmonised protection measures for the conservation of native forests in Northern, Central and Southern Burgenland, Vienna, Vienna Umland-South, Lower Austria South, Graz and Eastern Styria, Győr-Moson-Sopron, Vas and Zala counties (the so-called programme area). In the scope of this project, international cooperation between three project partners: the Austrian Research Centre for Forests (BFW, Austria), the Forest Research Institute – University of Sopron (SOE ERTI, Hungary) and the Vas County Government Office (VVÖH, Hungary) was established. Previous results and outputs of the SUSTREE project (Interreg Central Europe CE614), such as: a) Transnational delineation model of conservation and forest seed transfer zones in climate change, b) Report of intraspecific response function and derivation of climate transfer limits, SusSelect data, recommendations and c) Application of the species distribution models for the delineation of seed transfer zones/models in Central Europe, were put into practice during the project, focusing on two native deciduous forest tree species of the Austrian-Hungarian border region: European beech (Fagus sylvatica L.) and sessile oak (Quercus petraea (Matt.) Liebl.).During the REIN-Forest project, the following joint documents were prepared and several activities were implemented:1. Model-based document on the current state and future perspectives of European beech and sessile oak forests;2. Bilateral strategy for the transfer of forest reproductive material (FRM) and its use in the Austrian-Hungarian border region;3. Establishment of altogether six demonstration sites (three in each country) with local and climate-adapted FRM of European beech and sessile oak for long-term monitoring;4. Management and monitoring plan of the demonstration sites;5. Joint bilingual communication strategy, which included informative programmes and meetings with professionals, locals and schools and also education material for further use.REIN-Forest focused on using scientific results and outputs in the field of applied forestry and awareness-raising. Besides strategies, recommendations and reports that would facilitate forest managers' decisions for the future in the border region, events and workshops were offered for forestry practitioners, school pupils and the public and a short film and educational materials were published.

BackgroundPatient health data collected from a variety of nontraditional resources, commonly referred to as real-world data, can be a key information source for health and social science research. Social media platforms, such as Twitter (Twitter, Inc), offer vast amounts of real-world data. An important aspect of incorporating social media data in scientific research is identifying the demographic characteristics of the users who posted those data. Age and gender are considered key demographics for assessing the representativeness of the sample and enable researchers to study subgroups and disparities effectively. However, deciphering the age and gender of social media users poses challenges. ObjectiveThis scoping review aims to summarize the existing literature on the prediction of the age and gender of Twitter users and provide an overview of the methods used. MethodsWe searched 15 electronic databases and carried out reference checking to identify relevant studies that met our inclusion criteria: studies that predicted the age or gender of Twitter users using computational methods. The screening process was performed independently by 2 researchers to ensure the accuracy and reliability of the included studies. ResultsOf the initial 684 studies retrieved, 74 (10.8%) studies met our inclusion criteria. Among these 74 studies, 42 (57%) focused on predicting gender, 8 (11%) focused on predicting age, and 24 (32%) predicted a combination of both age and gender. Gender prediction was predominantly approached as a binary classification task, with the reported performance of the methods ranging from 0.58 to 0.96 F1-score or 0.51 to 0.97 accuracy. Age prediction approaches varied in terms of classification groups, with a higher range of reported performance, ranging from 0.31 to 0.94 F1-score or 0.43 to 0.86 accuracy. The heterogeneous nature of the studies and the reporting of dissimilar performance metrics made it challenging to quantitatively synthesize results and draw definitive conclusions. ConclusionsOur review found that although automated methods for predicting the age and gender of Twitter users have evolved to incorporate techniques such as deep neural networks, a significant proportion of the attempts rely on traditional machine learning methods, suggesting that there is potential to improve the performance of these tasks by using more advanced methods. Gender prediction has generally achieved a higher reported performance than age prediction. However, the lack of standardized reporting of performance metrics or standard annotated corpora to evaluate the methods used hinders any meaningful comparison of the approaches. Potential biases stemming from the collection and labeling of data used in the studies was identified as a problem, emphasizing the need for careful consideration and mitigation of biases in future studies. This scoping review provides valuable insights into the methods used for predicting the age and gender of Twitter users, along with the challenges and considerations associated with these methods.

Donor organ biomarkers with sufficient predictive value in liver transplantation (LT) are lacking. We herein evaluate liver viability and mitochondrial bioenergetics for their predictive capacity towards the outcome in LT. We enrolled 43 consecutive patients undergoing LT. Liver biopsy samples taken upon arrival after static cold storage were assessed by histology, real-time confocal imaging analysis (RTCA), and high-resolution respirometry (HRR) for mitochondrial respiration of tissue homogenates. Early allograft dysfunction (EAD) served as primary endpoint. HRR data were analysed with a focus on the efficacy of ATP production or P-L control efficiency, calculated as 1-L/P from the capacity of oxidative phosphorylation P and non-phosphorylating respiration L. Twenty-two recipients experienced EAD. Pre-transplant histology was not predictive of EAD. The mean RTCA score was significantly lower in the EAD cohort (−0.75 ± 2.27) compared to the IF cohort (0.70 ± 2.08; p = 0.01), indicating decreased cell viability. P-L control efficiency was predictive of EAD (0.76 ± 0.06 in IF vs. 0.70 ± 0.08 in EAD-livers; p = 0.02) and correlated with the RTCA score. Both RTCA and P-L control efficiency in biopsy samples taken during cold storage have predictive capacity towards the outcome in LT. Therefore, RTCA and HRR should be considered for risk stratification, viability assessment, and bioenergetic testing in liver transplantation.

The scyphozoan Cassiopea xamachana is an emerging cnidarian model system for studying regeneration, animal-algae symbiotic relationships, and various aspects of evolutionary biology including the early emergence of animal nervous systems. Cassiopea has a life cycle similar to other scyphozoans, which includes the alternation between a sessile, asexual form (polyp) and a sexually reproducing stage, the medusa. The transition between the two forms is called strobilation, where the polyp releases a miniature medusa, the iconic ephyra, that subsequently develops into the adult medusa. In addition, Cassiopea polyps may reproduce asexually by budding off free-swimming so-called planuloid buds. While the development of planuloid buds and polyps has been studied in some detail, little is known about the ontogeny of the sexually produced planula larva. Using immunofluorescence labeling and confocal microscopy, we examined neuromuscular development during metamorphosis of the planula larva into the juvenile polyp in C. xamachana. For this purpose, we used tyrosinated α-tubulin-, FMRFamide- and serotonin-like immunoreactivity together with phalloidin labeling. Our results show a planula nervous system that consists of a basiectodermal neural plexus with mostly longitudinally oriented neurites. This neural meshwork is connected to sensory neurons in the superficial stratum of the ectoderm, which are exclusively localized in the aboral half of the larva. During settlement, this aborally concentrated nervous system of the planula is replaced completely by the orally concentrated nervous system of the polyp. Adult polyps show an extensive nerve net with a loose concentration around the oral disc. These findings are consistent with data from other scyphozoans and most likely constitute a conserved feature of scyphozoan discomedusae. Taken together, the data currently available suggest an aborally concentrated nervous system including sensory cells as part of the neural ground pattern of cnidarian planula larvae. The reorganization of the nervous system from anterior to posterior in planula-to-polyp metamorphosis most likely also constitutes an ancestral trait in cnidarian evolution.

Abstract Normothermic machine perfusion (NMP) has emerged as an innovative organ preservation technique. Developing an understanding for the donor organ immune cell composition and its dynamic changes during NMP is essential. We aimed for a comprehensive characterization of immune cell (sub)populations, cell trafficking and cytokine release during liver NMP. Single-cell transcriptome profiling of human donor livers prior to, during NMP and after transplantation shows an abundance of CXC chemokine receptor 1+/2+ (CXCR1+/CXCR2+) neutrophils, which significantly decreased during NMP. This is paralleled by a large efflux of passenger leukocytes with neutrophil predominance in the perfusate. During NMP, neutrophils shift from a pro-inflammatory state towards an aged/chronically activated/exhausted phenotype, while anti-inflammatory/tolerogenic monocytes/macrophages are increased. We herein describe the dynamics of the immune cell repertoire, phenotypic immune cell shifts and a dominance of neutrophils during liver NMP, which potentially contribute to the inflammatory response. Our findings may serve as resource to initiate future immune-interventional studies.

Pneumocystis jirovecii, the fungal agent of human Pneumocystis pneumonia, is closely related to macaque Pneumocystis. Little is known about other Pneumocystis species in distantly related mammals, none of which are capable of establishing infection in humans. The molecular basis of host specificity in Pneumocystis remains unknown as experiments are limited due to an inability to culture any species in vitro. To explore Pneumocystis evolutionary adaptations, we have sequenced the genomes of species infecting macaques, rabbits, dogs and rats and compared them to available genomes of species infecting humans, mice and rats. Complete whole genome sequence data enables analysis and robust phylogeny, identification of important genetic features of the host adaptation, and estimation of speciation timing relative to the rise of their mammalian hosts. Our data reveals insights into the evolution of P. jirovecii, the sole member of the genus able to infect humans.