A Drosophila glial cell atlas reveals a mismatch between transcriptional and morphological diversity.

Morphology is a defining feature of neuronal identity. Like neurons, glia display diverse morphologies, both across and within glial classes, but are also known to be morphologically plastic. Here, we explored the relationship between glial morphology and transcriptional signature using the Drosophila central nervous system (CNS), where glia are categorised into 5 main classes (outer and inner surface glia, cortex glia, ensheathing glia, and astrocytes), which show within-class morphological diversity. We analysed and validated single-cell RNA sequencing data of Drosophila glia in 2 well-characterised tissues from distinct developmental stages, containing distinct circuit types: the embryonic ventral nerve cord (VNC) (motor) and the adult optic lobes (sensory). Our analysis identified a new morphologically and transcriptionally distinct surface glial population in the VNC. However, many glial morphological categories could not be distinguished transcriptionally, and indeed, embryonic and adult astrocytes were transcriptionally analogous despite differences in developmental stage and circuit type. While we did detect extensive within-class transcriptomic diversity for optic lobe glia, this could be explained entirely by glial residence in the most superficial neuropil (lamina) and an associated enrichment for immune-related gene expression. In summary, we generated a single-cell transcriptomic atlas of glia in Drosophila, and our extensive in vivo validation revealed that glia exhibit more diversity at the morphological level than was detectable at the transcriptional level. This atlas will serve as a resource for the community to probe glial diversity and function.

A story of vicariance? how the geology of oceanic archipelagos influenced the evolutionary history of endemic damselflies.

South Pacific islands provide an ideal study system to explore patterns of speciation, specifically examining the role of dispersal versus vicariance. Dispersal is often the suggested mechanism of diversification in the South Pacific, specifically among remote island chains. Here, we provide a phylogeny of several related genera of Coenagrionidae (Odonata: Zygoptera) from the South Pacific, based on five molecular loci, in order to examine patterns of speciation in the region. We used the endemic damselfly genera Nesobasis, Nikoulabasis, and Vanuatubasis found across both Fiji and Vanuatu. Knowledge of the geologic history of the region was used to inform our understanding of the evolution of these genera. Both archipelagos used to be part of the Vitiaz arc which spanned from the Solomon Islands to Tonga and began to break apart 10-12 Ma. Results of our divergence-time estimations and biogeographic reconstructions support that the breakup of this arc acted as a significant vicariance event in the evolution of these taxa. Specifically, it led to the extant generic diversity seen in these damselflies. We find that within the archipelago of Vanuatu, that Espiritu Santo served as an important source for dispersal to other islands with Malekula acting as a stepping stone to Efate.

Beetle bioluminescence outshines extant aerial predators.

We understand very little about the timing and origins of bioluminescence, particularly as a predator avoidance strategy. Understanding the timing of its origins, however, can help elucidate the evolution of this ecologically important signal. Using fireflies, a prevalent bioluminescent group where bioluminescence primarily functions as aposematic and sexual signals, we explore the origins of this signal in the context of their potential predators. Divergence time estimations were performed using genomic-scale datasets providing a robust estimate for the origin of firefly bioluminescence as both a terrestrial and as an aerial signal. Our results recover the origin of terrestrial beetle bioluminescence at 141.17 (122.63-161.17) Ma and firefly aerial bioluminescence at 133.18 (117.86-152.47) Ma using a large dataset focused on Lampyridae; and terrestrial bioluminescence at 148.03 (130.12-166.80) Ma, with the age of aerial bioluminescence at 104.97 (99.00-120.90) Ma using a complementary Elateroidea dataset. These ages pre-date the origins of all known extant aerial predators (i.e. bats and birds) and support much older terrestrial predators (assassin bugs, frogs, ground beetles, lizards, snakes, hunting spiders and harvestmen) as the drivers of terrestrial bioluminescence in beetles. These ages also support the hypothesis that sexual signalling was probably the original function of this signal in aerial fireflies.

Early Welsh cervical screening programme learning as a consequence of the coronavirus pandemic.

BACKGROUND: The coronavirus pandemic has had a profound impact on the cervical screening programme in Wales for the eligible women, sample takers, and laboratory and colposcopy services. AIMS: To explore what changes due to the pandemic have adversely affected screening outcomes in Wales and what lessons can be learned to improve cervical screening in Wales and elsewhere. METHODS: A review of the screening performance in 2020 and the first quarter of 2021 in Wales as well as a comparison with other cervical screening programme responses to the pandemic. RESULTS: A 3 month pause of screening together with a change in a variety of working practices, including social distancing, use of personal protective equipment, use of virtual meetings, and home working have been implemented. The combination of a pause to the issuing of invitations, plus reduced services in primary and secondary care, together with population lockdown, have contributed to longer waiting times for colposcopy and potentially delayed cancer diagnoses. Some programme changes which were being evaluated prior to the pandemic could be developed now to mitigate the impact of the pandemic such as improved information, increased screening intervals for human papillomavirus-based screening programmes, and home working for call and recall staff. CONCLUSIONS: Despite a considerable short-term interruption to the cervical screening programme, some changes introduced as a result of the coronavirus pandemic could provide key lessons learnt for improvement for cervical cancer prevention services.

Synergistic malaria vaccine combinations identified by systematic antigen screening.

A highly effective vaccine would be a valuable weapon in the drive toward malaria elimination. No such vaccine currently exists, and only a handful of the hundreds of potential candidates in the parasite genome have been evaluated. In this study, we systematically evaluated 29 antigens likely to be involved in erythrocyte invasion, an essential developmental stage during which the malaria parasite is vulnerable to antibody-mediated inhibition. Testing antigens alone and in combination identified several strain-transcending targets that had synergistic combinatorial effects in vitro, while studies in an endemic population revealed that combinations of the same antigens were associated with protection from febrile malaria. Video microscopy established that the most effective combinations targeted multiple discrete stages of invasion, suggesting a mechanistic explanation for synergy. Overall, this study both identifies specific antigen combinations for high-priority clinical testing and establishes a generalizable approach that is more likely to produce effective vaccines.

Missense mutations in ß-1,3-N-acetylglucosaminyltransferase 1 (B3GNT1) cause Walker-Warburg syndrome.

Several known or putative glycosyltransferases are required for the synthesis of laminin-binding glycans on alpha-dystroglycan (αDG), including POMT1, POMT2, POMGnT1, LARGE, Fukutin, FKRP, ISPD and GTDC2. Mutations in these glycosyltransferase genes result in defective αDG glycosylation and reduced ligand binding by αDG causing a clinically heterogeneous group of congenital muscular dystrophies, commonly referred to as dystroglycanopathies. The most severe clinical form, Walker-Warburg syndrome (WWS), is characterized by congenital muscular dystrophy and severe neurological and ophthalmological defects. Here, we report two homozygous missense mutations in the ß-1,3-N-acetylglucosaminyltransferase 1 (B3GNT1) gene in a family affected with WWS. Functional studies confirmed the pathogenicity of the mutations. First, expression of wild-type but not mutant B3GNT1 in human prostate cancer (PC3) cells led to increased levels of αDG glycosylation. Second, morpholino knockdown of the zebrafish b3gnt1 orthologue caused characteristic muscular defects and reduced αDG glycosylation. These functional studies identify an important role of B3GNT1 in the synthesis of the uncharacterized laminin-binding glycan of αDG and implicate B3GNT1 as a novel causative gene for WWS.

Jamb and jamc are essential for vertebrate myocyte fusion.

Cellular fusion is required in the development of several tissues, including skeletal muscle. In vertebrates, this process is poorly understood and lacks an in vivo-validated cell surface heterophilic receptor pair that is necessary for fusion. Identification of essential cell surface interactions between fusing cells is an important step in elucidating the molecular mechanism of cellular fusion. We show here that the zebrafish orthologues of JAM-B and JAM-C receptors are essential for fusion of myocyte precursors to form syncytial muscle fibres. Both jamb and jamc are dynamically co-expressed in developing muscles and encode receptors that physically interact. Heritable mutations in either gene prevent myocyte fusion in vivo, resulting in an overabundance of mononuclear, but otherwise overtly normal, functional fast-twitch muscle fibres. Transplantation experiments show that the Jamb and Jamc receptors must interact between neighbouring cells (in trans) for fusion to occur. We also show that jamc is ectopically expressed in prdm1a mutant slow muscle precursors, which inappropriately fuse with other myocytes, suggesting that control of myocyte fusion through regulation of jamc expression has important implications for the growth and patterning of muscles. Our discovery of a receptor-ligand pair critical for fusion in vivo has important implications for understanding the molecular mechanisms responsible for myocyte fusion and its regulation in vertebrate myogenesis.

Cachd1 interacts with Wnt receptors and regulates neuronal asymmetry in the zebrafish brain.

Neurons on the left and right sides of the nervous system often show asymmetric properties, but how such differences arise is poorly understood. Genetic screening in zebrafish revealed that loss of function of the transmembrane protein Cachd1 resulted in right-sided habenula neurons adopting left-sided identity. Cachd1 is expressed in neuronal progenitors, functions downstream of asymmetric environmental signals, and influences timing of the normally asymmetric patterns of neurogenesis. Biochemical and structural analyses demonstrated that Cachd1 can bind simultaneously to Lrp6 and Frizzled family Wnt co-receptors. Consistent with this, lrp6 mutant zebrafish lose asymmetry in the habenulae, and epistasis experiments support a role for Cachd1 in modulating Wnt pathway activity in the brain. These studies identify Cachd1 as a conserved Wnt receptor-interacting protein that regulates lateralized neuronal identity in the zebrafish brain.

Ground beetles suppress slugs in corn and soybean under conservation agriculture.

Conservation agriculture practices such as eliminating tillage and planting high residue cover crops are becoming increasingly important in field crop systems in the US Mid-Atlantic. However, these practices have sometimes been associated with an increase in moderate to severe damage to field crops by slugs. Conserving natural enemy populations is a desirable way to manage slug infestations because remedial control measures are limited. Here, we tested the effects of conservation practices, weather, and natural enemies on slug activity-density measured by tile traps placed among 41 corn and soybean fields during the spring of 2018 and 2019 in the Northern Shenandoah Valley, Virginia, USA. We found that a positive effect of cover crops on slug activity-density was reduced by tillage and that slug activity-density declined with increasing ground beetle activity-density. Slug activity-density also declined with decreasing rainfall and increasing average temperature. Weather was the only significant predictor of ground beetle activity-density, which was reduced in sites and weeks that were relatively hot and dry or that were cool and wet. However, we also found a marginally significant negative effect of pre-plant insecticides on ground beetles. We suggest that the observed interacting effects of cover crops and tillage reflect favorable conditions for slugs provided by increased small grain crop residue that can be mitigated to some extent by even low levels of tillage. More broadly, our study suggests that implementation of practices known to promote recruitment of ground beetles in crop fields can improve natural suppression of slugs in corn and soybean that are being increasingly cultivated according to conservation agriculture practices.

Characterising meaningful patient and public involvement in the pharmaceutical industry research setting: a retrospective quality assessment.

OBJECTIVES: Patient and public involvement (PPI) in clinical research has a well-established infrastructure in the UK, and while there has been good progress within pharmaceutical-industry-sponsored research, further improvements are still needed. This review aims to share learnings from quality assessments of historical PPI projects within Pfizer UK to inform future projects and drive PPI progress in the pharmaceutical industry. DESIGN AND SETTING: Internal assessments of Pfizer UK PPI projects were conducted to identify all relevant projects across the medicines development continuum between 2017 and 2021. Five sample projects were developed into case studies. OUTCOME MEASURE: Retrospective quality assessments were performed using the Patient Focused Medicines Development (PFMD) Patient Engagement Quality Guidance (PEQG) tool. Recommendations for improvement were developed. RESULTS: Retrospective case study analysis and quality framework assessment revealed benefits of PPI to both Pfizer UK and to external partners, as well as challenges and learnings to improve future practice. Recommendations for improvement based on these findings focused on processes and procedures for PPI, group dynamics and diversity for PPI activities, sharing of expertise, the importance of bidirectional and timely feedback, and the use of understandable language in materials. CONCLUSIONS: PPI in medicines development is impactful and beneficial but is still being optimised in the pharmaceutical industry. Using the PFMD PEQG tool to define gaps, share learnings and devise recommendations for improvement helps to ensure that PPI is genuine and empowering, rather than tokenistic. Ultimately, these recommendations should be acted on to further embed PPI as an integral part of medicines development and health research within the pharmaceutical industry. This article includes a plain language summary in the supplement.

Validating species distribution models to illuminate coastal fireflies in the South Pacific (Coleoptera: Lampyridae).

The coastal areas of Vanuatu are under a multitude of threats stemming from commercialization, human development, and climate change. Atyphella Olliff is a genus of firefly that includes species endemic to these coastal areas and will need protection. The research that has already been conducted was affected by accessibility due to the remote nature of the islands which left numerous knowledge gaps caused by a lack of distributional data (e.g., Wallacean shortfall). Species distribution models (SDM) are a powerful tool that allow for the modeling of the broader distribution of a taxon, even with limited distributional data available. SDMs assist in filling the knowledge gap by predicting potential areas that could contain the species of interest, making targeted collecting and conservation efforts more feasible when time, resources, and accessibility are major limiting factors. Here a MaxEnt prediction was used to direct field collecting and we now provide an updated predictive distribution for this endemic firefly genus. The original model was validated with additional fieldwork, ultimately expanding the known range with additional locations first identified using MaxEnt. A bias analysis was also conducted, providing insight into the effect that developments such as roads and settlements have on collecting and therefore the SDM, ultimately allowing for a more critical assessment of the overall model. After demonstrating the accuracy of the original model, this new updated SDM can be used to identify specific areas that will need to be the target of future conservation efforts by local government officials.

Carpophiline-ID: an interactive matrix-based key to the carpophiline sap beetles (Coleoptera, Nitidulidae) of Eastern North America.

Carpophiline-ID is presented, a matrix-based LucidTM key, for the adult stage of the known species of Carpophilinae (Coleoptera: Nitidulidae) of North America, east of the Mississippi River. An overview of the features and technical specifications used to build the key is provided. The list of terminal taxa used in the key represents the most current regional account for Carpophilinae, a beetle subfamily of agricultural and ecological importance. The value of matrix-based, free access keys for the identification of difficult taxa is discussed.

Factors Influencing the Distribution of Endemic Damselflies in Vanuatu.

Vanuatubasis Ober and Staniczek is a genus of damselfly endemic to Vanuatu. Little is known about the distribution and general natural history of the genus. We present the results of 14 weeks of fieldwork in Vanuatu to provide a better understanding of the biology of this genus. Specifically, we tested ecological niche models to predict the presence of Vanuatubasis throughout the region and explored how water pH may play a role in their distribution and ecology. The results of this fieldwork refined our model and further predicted the presence of this genus on additional islands. We also found stream pH as a strong predictor for the presence of Vanuatubasis, with their presence in alkaline streams significantly higher (p < 0.001). The mean pH for those streams where the genus was collected was 8.44 (n = 53).

A simple and effective F0 knockout method for rapid screening of behaviour and other complex phenotypes.

Hundreds of human genes are associated with neurological diseases, but translation into tractable biological mechanisms is lagging. Larval zebrafish are an attractive model to investigate genetic contributions to neurological diseases. However, current CRISPR-Cas9 methods are difficult to apply to large genetic screens studying behavioural phenotypes. To facilitate rapid genetic screening, we developed a simple sequencing-free tool to validate gRNAs and a highly effective CRISPR-Cas9 method capable of converting >90% of injected embryos directly into F0 biallelic knockouts. We demonstrate that F0 knockouts reliably recapitulate complex mutant phenotypes, such as altered molecular rhythms of the circadian clock, escape responses to irritants, and multi-parameter day-night locomotor behaviours. The technique is sufficiently robust to knockout multiple genes in the same animal, for example to create the transparent triple knockout crystal fish for imaging. Our F0 knockout method cuts the experimental time from gene to behavioural phenotype in zebrafish from months to one week.

Type designations for fireflies (Coleoptera: Lampyridae) of the Biologia Centrali Americana Gorham, 1881 housed in the Natural History Museum, London.

The Biologia Centrali Americana (B.C.A.) is comprised of eight volumes that deal specifically with Coleoptera. These volumes were split into 18 parts and were published between 1879 and 1911. The family Lampyridae was treated in two parts, the main text (1881) with a supplement (1884). Within volume three, part 2, Gorham lists ~90 species in 14 genera, not including the Phengodini subfamily. Of these, Gorham provided original descriptions for 37 species. During recent research visits (2018 and 2020) the authors were able to study material pertinent to the B.C.A. We were able to confidently designate holotypes, lectotypes, and paralectotypes following ICZN articles 73.1 and 74.1 within these species. Two species described by Gorham (1881) are not treated here. Phaenolis nirgricollis was located with a single specimen, already designate as the holotype. Two female syntypes of Photinus consanguineous were located, however Oliver (1907) synonymized these females with Photinus pyralis. These designations contribute to a larger taxonomic effort to stabilize the nomenclature of this group. The species described in the supplement will be treated in a future work. Subfamilies are listed according to Martin et al. (2019) and genera/species within each subfamily are listed according to the order in Gorham (1881).

Prevalence of leg regeneration in damselflies reevaluated: A case study in Coenagrionidae.

The leg regeneration capabilities of damselflies are understudied. Here we present the first data of regenerated limbs across a genus of damselfly based on adult specimens collected in the field to illustrate the prevalence of limb loss among nymphs. We show that this phenomenon is much more prevalent than previously thought, as 42 percent of individuals were found with regenerated limbs. Furthermore, we test for patterns within these data to begin to unravel the potential causes of limb loss in nymphal damselflies, showing that intrinsic factors such as sex and species cannot explain the patterns of limb loss pointing to environmental factors as the probable cause. We argue that Odonata limb regeneration provides a potentially unique perspective into the nymphal stage of these organisms.

Two new species of coastal Atyphella Olliff (Lampyridae: Luciolinae).

Additional work on the islands of Vanuatu has improved our understanding of the actual diversity of South Pacific coastal fireflies. Prior to recent fieldwork in Vanuatu, the only known lampyrid from Vanuatu was Atyphella aphrogeneia (Ballantyne), a coastal species also found in Papua New Guinea. After further examination, we determined that specimens from Vanuatu formerly classified as Atyphella aphrogeneia actually belong to an undescribed species. New species, Atyphella maritimus Saxton and Powell and Atyphella marigenous Saxton and Bybee, are described from specimens collected in Vanuatu. An updated key for coastal Atyphella in the South Pacific is provided.

Type designations for sap beetles in the subfamily Carpophilinae Erichson (Coleoptera: Nitidulidae) housed in the Natural History Museum, London.

The subfamily Carpophilinae, in particular the genus Carpophilus Stephens, represents one of the most speciose lineages within Nitidulidae. The subfamily was first recognized by Erichson (1843) in a much broader sense, including what is now the Cillaeinae. According to Kirejtshuk (2008), there are seven genera and 13 subgenera within Carpophilinae. To date, however, no comprehensive tribal or generic phylogenetic reconstructions have been published for the subfamily. This work constitutes a preliminary stage of a larger project that will address the subfamily in a rigorous taxonomic and systematic framework. A stable taxonomic foundation for Carpophilus species and other Carpophilinae taxa is needed to ensure the validity of future work in the subfamily. Herein we designate lectotypes and paralectotypes and confirm holotypes for 14 species of carpophiline sap beetles following ICZN (1999) articles 73.1 and 74.1.

Notes on the Carpophilus Stephens (Coleoptera: Nitidulidae) of Australia, with a new species from Victoria.

A new species of the genus Carpophilus Stephens is described from Australia. This species is currently placed in the subgenus Myothorax Murray, although the group is in need of formal revision. A diagnosis is given to distinguish the new species from all other species of Myothorax in Australia. Additionally, a discussion of type material from Carpophilus described by MacLeay is added with formal lectotype designations for Carpophilus aterrimus MacLeay and Carpophilus pilipennis MacLeay. Taxonomic notes are also given for Carpophilus aterrimus MacLeay, while Carpophilus planatus Murray is removed from synonymy and treated as a valid taxon.

Billbug (Coleoptera: Dryophthoridae: Sphenophorus spp.) Seasonal Biology and DNA-Based Life Stage Association in Indiana Turfgrass.

Eleven species of billbugs (Coleoptera: Dryophthoridae: Sphenophorus spp. Schönherr) infest managed turfgrass in North America. However, the regional variation in species composition remains unresolved and the seasonal phenology of several species has not been well documented. The latter gap is largely due to the inability to identify the larval stage to species-a confounding problem with several sympatric insect species. We used field trapping (adults) and soil sampling (larvae and pupae) surveys along with a DNA-based life-stage association to characterize the biology of billbugs associated with turfgrass in the Midwestern United States. Pitfall trapping at four locations in Indiana revealed four billbug species: S. venatus Say, S. parvulus Gyllenhaal, S. minimus Hart, and S. inaequalis Say. Sphenophorus venatus was the most abundant species on warm-season turfgrass while S. parvulus was most abundant on cool-season turfgrass. Investigation of S. venatus seasonal biology revealed two overwintered life stages-larva and adult-which resulted in two overlapping cohorts and two larval generations. Degree-day models describing S. venatus activity were more accurate for first-generation adults and larvae than for overwintering life stages. Maximum-likelihood analyses provided the first molecular species identification of billbug larvae and direct evidence that S. venatus larvae are capable of overwintering above 40°N latitude. Findings clarify the utility of molecular markers (CO1, 18S, and ITS2) for describing billbug larval population dynamics and seasonal phenology in regions where several sympatric billbug species occur. These results support the development of sustainable management strategies based on billbug seasonal phenology in different regions of North America.