Krüppel-like factor gene function in the ctenophore Mnemiopsis leidyi assessed by CRISPR/Cas9-mediated genome editing.

The Krüppel-like factor (Klf) gene family encodes transcription factors that play an important role in the regulation of stem cell proliferation, cell differentiation and development in bilaterians. Although Klf genes have been shown to specify functionally various cell types in non-bilaterian animals, their role in early-diverging animal lineages has not been assessed. Thus, the ancestral activity of these transcription factors in animal development is not well understood. The ctenophore Mnemiopsis leidyi has emerged as an important non-bilaterian model system for understanding early animal evolution. Here, we characterize the expression and functional role of Klf genes during M. leidyi embryogenesis. Zygotic Klf gene function was assessed with both CRISPR/Cas9-mediated genome editing and splice-blocking morpholino oligonucleotide knockdown approaches. Abrogation of zygotic Klf expression during M. leidyi embryogenesis resulted in abnormal development of several organs, including the pharynx, tentacle bulbs and apical organ. Our data suggest an ancient role for Klf genes in regulating endodermal patterning, possibly through regulation of cell proliferation.

Safety and Effectiveness of Large-Bore Percutaneous Cholangioscopy-Assisted Gallstone Retrieval for Inoperable Calculous Cholecystitis: A Multi-Institutional Retrospective Study.

PURPOSE: To evaluate the safety and effectiveness of large-bore percutaneous biliary access techniques for cholangioscopy-assisted gallstone extraction in patients with a history of acute calculous cholecystitis who are poor surgical candidates. MATERIALS AND METHODS: A retrospective analysis was conducted on patients who underwent percutaneous cholangioscopy for gallstone extraction using large-bore access (24 or 30 French) at two large academic centers from September 2020 and August 2022. Technical success, procedure duration, fluoroscopy time, immediate post-procedure symptom reduction, 3-month symptom-free outcomes, and adverse effects were assessed. RESULTS: Thirty consecutive patients were included. Gallstone removal in a single cholangioscopy session was successful in 93.3% of cases. Large-bore access facilitated the removal of gallstones ranging from 0.5 to 4 cm in diameter, with mean procedure and fluoroscopy times of 105.4 mins 21.7 mins, respectively. This compares to a previous small cholangioscopy study of 13 patients demonstrating mean procedure and fluoroscopy times of 164 mins and 30 mins, respectively. All patients who presented for 3-month follow-up remained symptom-free without gallstone recurrence on imaging. The overall adverse event rate was 6.7%, one Grade C and one Grade D based on the SIR adverse events grading system, both managed appropriately, leading to patient discharge home. CONCLUSION: Large-bore percutaneous biliary access for cholangioscopy-assisted gallstone extraction is a safe and effective technique for managing symptomatic cholelithiasis in poor surgical candidates. Further prospective studies are warranted to validate these findings and evaluate long-term treatment outcomes.

The effects of family, culture and sex on linguistic development across 20 languages.

Languages vary in their complexity; caregivers vary in the way they structure their communicative interactions with children; and boys and girls can differ in their language skills. Using a multilevel modelling approach, we explored how these factors influence the path of language acquisition for young children growing up around the world (mean age 2-years 9-months; 56 girls). Across 43 different sites, we analysed 103 mother-child pairs who spoke 3,170,633 utterances, 16,209,659 morphemes, divided across 20 different languages: Afrikaans, Catalan, Cantonese, Danish, Dutch, English, Farsi, French, German, Hebrew, Icelandic, Irish, Italian, Japanese, Mandarin, Norwegian, Portuguese, Spanish, Swedish and Turkish. Using mean length of utterance (MLU) as a measure of language complexity and developmental skill, we found that variation in children's MLU was significantly explained by (a) between-language differences; namely the rate of child MLU growth was attuned to the complexity of their mother tongue, and (b) between-mother differences; namely mothers who used higher MLUs tended to have children with higher MLUs, regardless of which language they were learning and especially in the very young (<2.5 years-old). Controlling for family and language environment, we found no evidence of MLU sex differences in child speech nor in the speech addressed to boys and girls. By modelling language as a multilevel structure with cross-cultural variation, we were able to disentangle those factors that make children's pathway to language different and those that make it alike. RESEARCH HIGHLIGHTS: The speech of 103 mother-child pairs from 20 different languages showed large variation in the path of early language development. Language, family, but not the sex of the child, accounted for a significant proportion of individual differences in child speech, especially in the very young. The rate at which children learned language was attuned to the complexity of their mother tongue, with steeper trajectories for more complex language. Results demonstrate the relative influence of culture, family, and sex in shaping the path of language acquisition for different children.

Invasion genomics uncover contrasting scenarios of genetic diversity in a widespread marine invader.

Invasion rates have increased in the past 100 y irrespective of international conventions. What characterizes a successful invasion event? And how does genetic diversity translate into invasion success? Employing a whole-genome perspective using one of the most successful marine invasive species world-wide as a model, we resolve temporal invasion dynamics during independent invasion events in Eurasia. We reveal complex regionally independent invasion histories including cases of recurrent translocations, time-limited translocations, and stepping-stone range expansions with severe bottlenecks within the same species. Irrespective of these different invasion dynamics, which lead to contrasting patterns of genetic diversity, all nonindigenous populations are similarly successful. This illustrates that genetic diversity, per se, is not necessarily the driving force behind invasion success. Other factors such as propagule pressure and repeated introductions are an important contribution to facilitate successful invasions. This calls into question the dominant paradigm of the genetic paradox of invasions, i.e., the successful establishment of nonindigenous populations with low levels of genetic diversity.

The Role of Extracellular Vesicles in Allergic Sensitization: A Systematic Review.

Allergies affect approximately 10-30% of people worldwide, with an increasing number of cases each year; however, the underlying mechanisms are still poorly understood. In recent years, extracellular vesicles (EVs) have been suggested to play a role in allergic sensitization and skew to a T helper type 2 (Th2) response. The aim of this review is to highlight the existing evidence of EV involvement in allergies. A total of 22 studies were reviewed; 12 studies showed EVs can influence a Th2 response, while 10 studies found EVs promoted a Th1 or Treg response. EVs can drive allergic sensitization through up-regulation of pro-Th2 cytokines, such as IL-4 and IL-13. In addition, EVs from MRSA can induce IgE hypersensitivity in mice towards MRSA. On the other hand, EVs can induce tolerance in the immune system; for example, pre-exposing OVA-loaded EVs prevented OVA sensitization in mice. The current literature thus suggests that EVs play an essential role in allergy. Further research utilizing human in vitro models and clinical studies is needed to give a reliable account of the role of EVs in allergy.

The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research.

Reproducible science requires transparent reporting. The ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments) were originally developed in 2010 to improve the reporting of animal research. They consist of a checklist of information to include in publications describing in vivo experiments to enable others to scrutinise the work adequately, evaluate its methodological rigour, and reproduce the methods and results. Despite considerable levels of endorsement by funders and journals over the years, adherence to the guidelines has been inconsistent, and the anticipated improvements in the quality of reporting in animal research publications have not been achieved. Here, we introduce ARRIVE 2.0. The guidelines have been updated and information reorganised to facilitate their use in practice. We used a Delphi exercise to prioritise and divide the items of the guidelines into 2 sets, the "ARRIVE Essential 10," which constitutes the minimum requirement, and the "Recommended Set," which describes the research context. This division facilitates improved reporting of animal research by supporting a stepwise approach to implementation. This helps journal editors and reviewers verify that the most important items are being reported in manuscripts. We have also developed the accompanying Explanation and Elaboration (E&E) document, which serves (1) to explain the rationale behind each item in the guidelines, (2) to clarify key concepts, and (3) to provide illustrative examples. We aim, through these changes, to help ensure that researchers, reviewers, and journal editors are better equipped to improve the rigour and transparency of the scientific process and thus reproducibility.

Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0.

Improving the reproducibility of biomedical research is a major challenge. Transparent and accurate reporting is vital to this process; it allows readers to assess the reliability of the findings and repeat or build upon the work of other researchers. The ARRIVE guidelines (Animal Research: Reporting In Vivo Experiments) were developed in 2010 to help authors and journals identify the minimum information necessary to report in publications describing in vivo experiments. Despite widespread endorsement by the scientific community, the impact of ARRIVE on the transparency of reporting in animal research publications has been limited. We have revised the ARRIVE guidelines to update them and facilitate their use in practice. The revised guidelines are published alongside this paper. This explanation and elaboration document was developed as part of the revision. It provides further information about each of the 21 items in ARRIVE 2.0, including the rationale and supporting evidence for their inclusion in the guidelines, elaboration of details to report, and examples of good reporting from the published literature. This document also covers advice and best practice in the design and conduct of animal studies to support researchers in improving standards from the start of the experimental design process through to publication.

Multi-institutional Retrospective Study of Percutaneous Cholangioscopy-Assisted Lithotripsy for Inoperable Calculous Cholecystitis.

PURPOSE: This study aimed to assess the safety and efficacy of percutaneous lithotripsy for gallstone eradication in patients with calculous cholecystitis with stones >1 cm. MATERIALS AND METHODS: Multi-institutional institutional review board approved retrospective review of patients who presented with calculous cholecystitis and were not determined to be surgical candidates. All patients underwent percutaneous cholecystostomy tube placement for acute infection, which was later exchanged for a large sheath for ShockPulse (Olympus, Tokyo, Japan) lithotripsy and stone destruction. Review parameters included procedural technical and clinical data, including clinical presentation, mean length of hospital stay, and postintervention symptom reduction. RESULTS: Twelve patients (mean age, 74.6 years; range, 52-94 years; 6 men and 6 women) underwent large-bore sheath (24-30 F) cholangioscopy-assisted gallstone destruction via rigid lithotripsy. The size of the gallstones ranged from 1.2 to 4.0 cm. All patients had prior cholecystostomy access for a mean of 25 weeks before gallstone extraction to ensure tract maturation via transhepatic or transperitoneal access. The technical success rate in single-session stone removal was 100%, with no major procedure-related adverse events. All patients were symptom- and pain-free after the procedure. The mean procedure time was 111.5 minutes, and the mean fluoroscopy time was 19.2 minutes. The median length of hospital stay was 1 day after the procedure. The mean time from percutaneous lithotripsy to biliary tube removal was 35 days (range, 17-45 days). CONCLUSIONS: Fluoroscopy-guided percutaneous rigid lithotripsy is a safe and effective procedure for gallstone destruction and extraction in patients who are poor surgical candidates with large lumen-occupying cholelithiasis.

Metabolic precision labeling enables selective probing of O-linked N-acetylgalactosamine glycosylation.

Protein glycosylation events that happen early in the secretory pathway are often dysregulated during tumorigenesis. These events can be probed, in principle, by monosaccharides with bioorthogonal tags that would ideally be specific for distinct glycan subtypes. However, metabolic interconversion into other monosaccharides drastically reduces such specificity in the living cell. Here, we use a structure-based design process to develop the monosaccharide probe N-(S)-azidopropionylgalactosamine (GalNAzMe) that is specific for cancer-relevant Ser/Thr(O)-linked N-acetylgalactosamine (GalNAc) glycosylation. By virtue of a branched N-acylamide side chain, GalNAzMe is not interconverted by epimerization to the corresponding N-acetylglucosamine analog by the epimerase N-acetylgalactosamine-4-epimerase (GALE) like conventional GalNAc-based probes. GalNAzMe enters O-GalNAc glycosylation but does not enter other major cell surface glycan types including Asn(N)-linked glycans. We transfect cells with the engineered pyrophosphorylase mut-AGX1 to biosynthesize the nucleotide-sugar donor uridine diphosphate (UDP)-GalNAzMe from a sugar-1-phosphate precursor. Tagged with a bioorthogonal azide group, GalNAzMe serves as an O-glycan-specific reporter in superresolution microscopy, chemical glycoproteomics, a genome-wide CRISPR-knockout (CRISPR-KO) screen, and imaging of intestinal organoids. Additional ectopic expression of an engineered glycosyltransferase, "bump-and-hole" (BH)-GalNAc-T2, boosts labeling in a programmable fashion by increasing incorporation of GalNAzMe into the cell surface glycoproteome. Alleviating the need for GALE-KO cells in metabolic labeling experiments, GalNAzMe is a precision tool that allows a detailed view into the biology of a major type of cancer-relevant protein glycosylation.

Affective trajectories: Are hens influenced by positive and negative changes in their living conditions?

Most studies of the effects of housing and husbandry on animals' affective states and welfare investigate the impact of stable living conditions, comparing for example, animals living in enriched environments with those living in non-enriched ones. Changes in living conditions, including from more to less enriched environments, have also been found to have effects on measures of affective state and welfare in some species. But these studies have not investigated whether it is the trajectory of change that has affected the animals (e.g., worsening conditions), or simply the nature of their final environment (e.g., non-enriched). Here, we hypothesised that laying hens living in worsening conditions across a six-week period (gradually moving from preferred to non-preferred living conditions; "Trajectory to Non-Preferred", TNP, n = 30), would show evidence of more negative affective states and poorer welfare than those living continuously in non-preferred conditions for the same duration ("Stable Non-Preferred", SNP, n = 30). We also hypothesised that hens living in improving conditions (gradually moving from non-preferred to preferred living conditions; "Trajectory to Preferred", TP, n = 30), would show evidence of more positive affective states and better welfare than those living continuously in preferred conditions ("Stable Preferred", SP, n = 30). The preferred living condition provided extensive resources and intermittent rewarding events (such as the delivery of food treats) known to be valued and preferred by most hens, while the non-preferred living condition provided just basic resources and intermittent aversive events (e.g., loud noises). The hens' affective states and welfare were measured using home-pen behavioural observations, body condition assessments, physiological stress measures (e.g., blood corticosterone, glucose, etc.), physical challenge tests, and judgement bias tests. A number of differences between hens in the trajectory and stable living conditions were found: TP hens were lighter, showed more foraging behaviour and less standing alert and head-shaking than SP hens, while TNP hens showed more head-shaking, mild feather pecking and aggressive attacking of pen mates than SNP hens. However, some of these differences failed to reach significance following Benjamini adjustments for multiple testing. The groups also did not differ in their judgement biases (measured in a sub-sample of 12 hens per experimental group), response to physical challenges, or measures of physiological stress. We conclude that the hens in the present study showed some evidence of responsiveness to 'affective trajectories' in their living conditions, but no definitive effects on their affective states and welfare.

Depression and thoughts of self-harm and suicide among people living with dementia: results of a cross-sectional survey.

BACKGROUND: Depression is common among people with dementia. Despite most people with dementia living in the community, there have been few investigations of self-reported depressive symptoms and suicidal ideation among community-dwelling people with dementia in Australia. This study aimed to explore the proportion of people with mild, moderate and severe levels of depressive symptoms, and suicidal ideation among a sample of people living with dementia in Australia. Correlates of reporting depressive symptoms were also explored. METHODS: Adults diagnosed with dementia by a medical professional who were English speaking and community-dwelling were asked to complete a paper and pencil survey. Those who were unable to provide independent consent were excluded. Depression was assessed using the Geriatric Depression Scale -15, and suicidal ideation was assessed using two study-specific items. Multivariable analyses examined quality of life, unmet needs and sociodemographic factors associated with having a score of five or more on the Geriatric Depression Scale-15. RESULTS: Ninety-four people participated in the study. Thirty-seven percent (n = 35) reported some level of depressive symptoms, with most of these (21%, n = 20) classified as having mild depressive symptoms. Five participants (5%) reported they had had thoughts of being better off dead or hurting themselves, while three (3%) reported having had a plan to end their life. For each additional unmet need, the odds of being depressed increased by 25%(P < 0.001). For each point increase in quality of life, the odds of being depressed decreased by 48% (P < 0.001). CONCLUSIONS: The high proportion of people with dementia who report depressive symptoms suggests the need to routinely assess depressive symptoms among this group. There may also be benefits in assessing unmet needs and addressing these where possible as part of an approach to reducing depression among people living with dementia in the community.

The Early Humor Survey (EHS): A reliable parent-report measure of humor development for 1- to 47-month-olds.

We created a 20-item parent-report measure of humor development from 1 to 47 months: the Early Humor Survey (EHS). We developed the EHS with Study 1 (N = 219) using exploratory factor analysis, demonstrating the EHS works with 1- to 47-month-olds with excellent reliability and a strong correlation with age, showing its developmental trajectory. We replicated the EHS with Study 2 (N = 587), revealing a one-factor structure, showing excellent reliability, and replicating a strong correlation with age. Study 3 (N = 84) found the EHS correlated with a humor experiment, however it no longer correlated once age was accounted for, suggesting low convergent validity. Subsamples of parents from Studies 2 and 3 showed excellent inter-observer reliability between both parents, and good longitudinal stability after 6 months. Combining participants from all studies, we found the EHS is reliable across countries (Australia, United Kingdom, United States), parent education levels, and children's age groups. We charted expected humor development by age (in months), and the expected proportion of children who would appreciate each humor type by age (in months). Finally, we found no demographic differences (e.g., country: Australia, Canada, United Kingdom, United States; parents' education) in humor when pooling all data. The EHS is a valuable tool that will allow researchers to understand how humor: (1) emerges; and (2) affects other aspects of life, e.g., making friends, coping with stress, and creativity. The EHS is helpful for parents, early years educators, and children's media, as it systematically charts early humor development.

The Early Social Cognition Inventory (ESCI): An examination of its psychometric properties from birth to 47 months.

Social cognition refers to a broad range of cognitive processes and skills that allow individuals to interact with and understand others, including a variety of skills from infancy through preschool and beyond, e.g., joint attention, imitation, and belief understanding. However, no measures examine socio-cognitive development from birth through preschool. Current test batteries and parent-report measures focus either on infancy, or toddlerhood through preschool (and beyond). We report six studies in which we developed and tested a new 21-item parent-report measure of social cognition targeting 0-47 months: the Early Social Cognition Inventory (ESCI). Study 1 (N = 295) revealed the ESCI has excellent internal reliability, and a two-factor structure capturing social cognition and age. Study 2 (N = 605) also showed excellent internal reliability and confirmed the two-factor structure. Study 3 (N = 84) found a medium correlation between the ESCI and a researcher-administered social cognition task battery. Study 4 (N = 46) found strong 1-month test-retest reliability. Study 5 found longitudinal stability (6 months: N = 140; 12 months: N = 39), and inter-observer reliability between parents (N = 36) was good, and children's scores increased significantly over 6 and 12 months. Study 6 showed the ESCI was internally reliable within countries (Australia, Canada, United Kingdom, United States, Trinidad and Tobago); parent ethnicity; parent education; and age groups from 4-39 months. ESCI scores positively correlated with household income (UK); children with siblings had higher scores; and Australian parents reported lower scores than American, British, and Canadian parents.

The ARRIVE guidelines 2.0: updated guidelines for reporting animal research.

Reproducible science requires transparent reporting. The ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments) were originally developed in 2010 to improve the reporting of animal research. They consist of a checklist of information to include in publications describing in vivo experiments to enable others to scrutinise the work adequately, evaluate its methodological rigour, and reproduce the methods and results. Despite considerable levels of endorsement by funders and journals over the years, adherence to the guidelines has been inconsistent, and the anticipated improvements in the quality of reporting in animal research publications have not been achieved. Here, we introduce ARRIVE 2.0. The guidelines have been updated and information reorganised to facilitate their use in practice. We used a Delphi exercise to prioritise and divide the items of the guidelines into 2 sets, the 'ARRIVE Essential 10,' which constitutes the minimum requirement, and the 'Recommended Set,' which describes the research context. This division facilitates improved reporting of animal research by supporting a stepwise approach to implementation. This helps journal editors and reviewers verify that the most important items are being reported in manuscripts. We have also developed the accompanying Explanation and Elaboration document, which serves (1) to explain the rationale behind each item in the guidelines, (2) to clarify key concepts, and (3) to provide illustrative examples. We aim, through these changes, to help ensure that researchers, reviewers, and journal editors are better equipped to improve the rigour and transparency of the scientific process and thus reproducibility.

The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research.

Reproducible science requires transparent reporting. The ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments) were originally developed in 2010 to improve the reporting of animal research. They consist of a checklist of information to include in publications describing in vivo experiments to enable others to scrutinise the work adequately, evaluate its methodological rigour, and reproduce the methods and results. Despite considerable levels of endorsement by funders and journals over the years, adherence to the guidelines has been inconsistent, and the anticipated improvements in the quality of reporting in animal research publications have not been achieved. Here, we introduce ARRIVE 2.0. The guidelines have been updated and information reorganised to facilitate their use in practice. We used a Delphi exercise to prioritise and divide the items of the guidelines into 2 sets, the "ARRIVE Essential 10," which constitutes the minimum requirement, and the "Recommended Set," which describes the research context. This division facilitates improved reporting of animal research by supporting a stepwise approach to implementation. This helps journal editors and reviewers verify that the most important items are being reported in manuscripts. We have also developed the accompanying Explanation and Elaboration document, which serves (1) to explain the rationale behind each item in the guidelines, (2) to clarify key concepts, and (3) to provide illustrative examples. We aim, through these changes, to help ensure that researchers, reviewers, and journal editors are better equipped to improve the rigour and transparency of the scientific process and thus reproducibility.

The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research.

Reproducible science requires transparent reporting. The ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments) were originally developed in 2010 to improve the reporting of animal research. They consist of a checklist of information to include in publications describing in vivo experiments to enable others to scrutinise the work adequately, evaluate its methodological rigour, and reproduce the methods and results. Despite considerable levels of endorsement by funders and journals over the years, adherence to the guidelines has been inconsistent, and the anticipated improvements in the quality of reporting in animal research publications have not been achieved. Here, we introduce ARRIVE 2.0. The guidelines have been updated and information reorganised to facilitate their use in practice. We used a Delphi exercise to prioritise and divide the items of the guidelines into 2 sets, the "ARRIVE Essential 10," which constitutes the minimum requirement, and the "Recommended Set," which describes the research context. This division facilitates improved reporting of animal research by supporting a stepwise approach to implementation. This helps journal editors and reviewers verify that the most important items are being reported in manuscripts. We have also developed the accompanying Explanation and Elaboration document, which serves (1) to explain the rationale behind each item in the guidelines, (2) to clarify key concepts, and (3) to provide illustrative examples. We aim, through these changes, to help ensure that researchers, reviewers, and journal editors are better equipped to improve the rigour and transparency of the scientific process and thus reproducibility.

Experimentally manipulating light spectra reveals the importance of dark corridors for commuting bats.

The rapid global spread of artificial light at night is causing unprecedented disruption to ecosystems. In otherwise dark environments, street lights restrict the use of major flight routes by some bats, including the threatened lesser horseshoe bat Rhinolophus hipposideros, and may disrupt foraging. Using radio tracking, we examined the response of individual female R. hipposideros to experimental street lights placed on hedgerows used as major flight routes. Hedgerows were illuminated on one side over four nights using lights with different emission spectra, while the opposite side of the hedge was not illuminated. Automated bat detectors were used to examine changes in overall bat activity by R. hipposideros and other bat species present. R. hipposideros activity reduced significantly under all light types, including red light, challenging a previously held assumption that red light is safe for bats. Despite this, R. hipposideros rapidly adapted to the presence of lights by switching their flight paths to the dark side of the hedgerow, enabling them to reach foraging sites without restriction. Red light had no effect on the activity of the other species present. Slow-flying Myotis spp. avoided orange, white and green light, while more agile Pipistrellus spp. were significantly more active at these light types compared to dark controls, most probably in response to accumulations of insect prey. No effect of any light type was found for Nyctalus or Eptesicus spp. Our findings demonstrate that caution must be used when promoting forms of lighting that are thought to be safe for wildlife before they are tested more widely. We argue that it is essential to preserve dark corridors to mitigate the impacts of artificial light at night on bat activity and movements.

State-dependent judgement bias in Drosophila: evidence for evolutionarily primitive affective processes.

Affective states influence decision-making under ambiguity in humans and other animals. Individuals in a negative state tend to interpret ambiguous cues more negatively than individuals in a positive state. We demonstrate that the fruit fly, Drosophila melanogaster, also exhibits state-dependent changes in cue interpretation. Drosophila were trained on a Go/Go task to approach a positive (P) odour associated with a sugar reward and actively avoid a negative (N) odour associated with shock. Trained flies were then either shaken to induce a purported negative state or left undisturbed (control), and given a choice between: air or P; air or N; air or ambiguous odour (1 : 1 blend of P : N). Shaken flies were significantly less likely to approach the ambiguous odour than control flies. This 'judgement bias' may be mediated by changes in neural activity that reflect evolutionarily primitive affective states. We cannot say whether such states are consciously experienced, but use of this model organism's versatile experimental tool kit may facilitate elucidation of their neural and genetic basis.

Comprehensive analysis of Hox gene expression in the amphipod crustacean Parhyale hawaiensis.

Hox genes play crucial roles in establishing regional identity along the anterior-posterior axis in bilaterian animals, and have been implicated in generating morphological diversity throughout evolution. Here we report the identification, expression, and initial genomic characterization of the complete set of Hox genes from the amphipod crustacean Parhyale hawaiensis. Parhyale is an emerging model system that is amenable to experimental manipulations and evolutionary comparisons among the arthropods. Our analyses indicate that the Parhyale genome contains a single copy of each canonical Hox gene with the exception of fushi tarazu, and preliminary mapping suggests that at least some of these genes are clustered together in the genome. With few exceptions, Parhyale Hox genes exhibit both temporal and spatial colinearity, and expression boundaries correlate with morphological differences between segments and their associated appendages. This work represents the most comprehensive analysis of Hox gene expression in a crustacean to date, and provides a foundation for functional studies aimed at elucidating the role of Hox genes in arthropod development and evolution.

The maternal-zygotic transition and zygotic activation of the Mnemiopsis leidyi genome occurs within the first three cleavage cycles.

The maternal-zygotic transition (MZT) describes the developmental reprogramming of gene expression marked by the degradation of maternally supplied gene products and activation of the zygotic genome. While the timing and duration of the MZT vary among taxa, little is known about early-stage transcriptional dynamics in the non-bilaterian phylum Ctenophora. We sought to better understand the extent of maternal mRNA loading and subsequent differential transcript abundance during the earliest stages of development by performing comprehensive RNA-sequencing-based analyses of mRNA abundance in single- and eight-cell stage embryos in the lobate ctenophore Mnemiopsis leidyi. We found 1,908 contigs with significant differential abundance between single- and eight-cell stages, of which 1,208 contigs were more abundant at the single-cell stage and 700 contigs were more abundant at the eight-cell stage. Of the differentially abundant contigs, 267 were exclusively present in the eight-cell samples, providing strong evidence that both the MZT and zygotic genome activation (ZGA) have commenced by the eight-cell stage. Many highly abundant transcripts encode genes involved in molecular mechanisms critical to the MZT, such as maternal transcript degradation, serine/threonine kinase activity, and chromatin remodeling. Our results suggest that chromosomal restructuring, which is critical to ZGA and the initiation of transcriptional regulation necessary for normal development, begins by the third cleavage within 1.5 hr post-fertilization in M. leidyi.