Knocking out barriers to engineered cell activity.

CRISPR-Cas9 gene-edited T cells show safety and long-term engraftment in human

Trypanosomes are monophyletic: evidence from genes for glyceraldehyde phosphate dehydrogenase and small subunit ribosomal RNA.

The genomes of Trypanosoma brucei, Trypanosoma cruzi and Leishmania major have been sequenced, but the phylogenetic relationships of these three protozoa remain uncertain. We have constructed trypanosomatid phylogenies based on genes for glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) and small subunit ribosomal RNA (SSU rRNA). Trees based on gGAPDH nucleotide and amino acid sequences (51 taxa) robustly support monophyly of genus Trypanosoma, which is revealed to be a relatively late-evolving lineage of the family Trypanosomatidae. Other trypanosomatids, including genus Leishmania, branch paraphyletically at the base of the trypanosome clade. On the other hand, analysis of the SSU rRNA gene data produced equivocal results, as trees either robustly support or reject monophyly depending on the range of taxa included in the alignment. We conclude that the SSU rRNA gene is not a reliable marker for inferring deep level trypanosome phylogeny. The gGAPDH results support the hypothesis that trypanosomes evolved from an ancestral insect parasite, which adapted to a vertebrate/insect transmission cycle. This implies that the switch from terrestrial insect to aquatic leech vectors for fish and some amphibian trypanosomes was secondary. We conclude that the three sequenced pathogens, T. brucei, T. cruzi and L. major, are only distantly related and have distinct evolutionary histories

The Interaction of Retention, Recruitment, and Density-Dependent Mortality in the Spatial Placement of Marine Reserves

Population density can affect rates of mortality and individual growth. We measured these for the non-exploited bluehead wrasse, Thalassoma bifasciatum, at three sites around St. Croix, US Virgin Islands. Previous work demonstrated that differences in the degree of larval retention in these sites results in very large differences in recruitment intensity. Post-settlement mortality differed among sites and was positively related to recruitment density. Post-settlement growth differences were small. Because of strong mortality effects early in life, adult densities and size/age distributions differed among sites and did not reflect differences in recruitment rate. The site with the highest retention and recruitment (Butler Bay) had many small fish, while the two other sites with lower recruitment rates (Jacks Bay and Green Cay) had proportionally more large fish. These differences resulted in large differences in egg production. Per capita production was highest at the lowest density site (Green Cay). Total egg production at Green Cay was 75% that at Butler Bay, despite only having half the population size, and the highest overall production was at Jacks Bay, with low retention and moderate recruitment. In terms of marine reserve location, sites predicted to have high retention and recruitment may not always be the sites of highest egg production due to density-dependent processes, and it is important to consider the relative values of self-recruitment and larval export in reserve design

Imaging Modalities Relevant to Intracranial Pressure Assessment in Astronauts: A Case-Based Discussion

Introduction: Intracranial pressure (ICP) elevation has been inferred or documented in a number of space crewmembers. Recent advances in noninvasive imaging technology offer new possibilities for ICP assessment. Most International Space Station (ISS) partner agencies have adopted a battery of occupational health monitoring tests including magnetic resonance imaging (MRI) pre- and postflight, and high-resolution sonography of the orbital structures in all mission phases including during flight. We hypothesize that joint consideration of data from the two techniques has the potential to improve quality and continuity of crewmember monitoring and care. Methods: Specially designed MRI and sonographic protocols were used to image eyes and optic nerves (ON) including the meningeal sheaths. Specific crewmembers multi-modality imaging data were analyzed to identify points of mutual validation as well as unique features of complementary nature. Results and Conclusion: Magnetic resonance imaging (MRI) and high-resolution sonography are both tomographic methods, however images obtained by the two modalities are based on different physical phenomena and use different acquisition principles. Consideration of the images acquired by these two modalities allows cross-validating findings related to the volume and fluid content of the ON subarachnoid space, shape of the globe, and other anatomical features of the orbit. Each of the imaging modalities also has unique advantages, making them complementary techniques

Noninvasive Techniques for Intracranial Pressure Assessment: A Review from Aerospace Medicine Perspective

Microgravity-induced changes in fluid distribution and other physiological factors due to space flight have been implicated as the cause of increased intracranial pressure (ICP) in a number of space crewmembers. The modest levels of ICP elevation and absence of severe symptoms in this group do not warrant invasive diagnostic interventions. However, the long-term trends and residual or consequential changes secondary to the observed ICP elevation in this group are not yet known. Therefore, close attention is needed to evaluate the potential techniques of noninvasively assessing ICP, including those feasible for in-flight use. Of particular interest is continuity between ground and in-flight testing, whereby data from the same or different techniques allow reasonably dependable estimation of ICP trends and responses. Methods: A thorough review of current literature, analysis of NASA data, and interviews with subject matter experts were conducted to construct a presentation that reflects the state of the art for noninvasive ICP measurement and monitoring. Results: Multiple imaging and non-imaging modalities are available to assess ICP in terrestrial clinical and experimental environments. Imaging alternatives include magnetic resonance imaging (MRI) and high-resolution sonography. Non-imaging techniques include transcranial Doppler, certain audiological methods, and venous ophthalmodynamometry, among others. Special functional techniques have been proposed recently that allow the use of advanced MRI methods to calculate ICP in addition to the acquisition of high-resolution images. Our data include many of these applications, with several cases of correlation with lumbar puncture, the invasive "gold standard" measurement of ICP

Points and Pits: Archaeological Investigations in Minnesota’s Region 9, the Lake Superior Shore, Carlton, Cook, Lake, and St. Louis Counties, Minnesota

Duluth Archaeology Center Report No. 11-22. Duluth: Duluth Archeology Center.This report is essentially an archeological survey of northeastern Minnesota’s coastal region, for which little is known about prehistoric human occupation. The authors first conducted a literature review, then carried out field investigations at 34 sites in 2010. Most of the sites were already known, and only four new sites suggesting historic use or habitation were found, all in Jay Cooke State Park. The study focused on sites that were not far from water. The report describes archeological and GIS methods used to select and explore the sites. The authors conclude that some progress has been made in contributing to a historical database for the region, but that much more remains to be discovered about prehistoric occupation on the Lake Superior shoreline. Key sections of the report are extracted and reproduced below. Summary: "This project was designed to add to our knowledge of Region 9, which was poorly known as a result of relatively few previously recorded. Four objectives were stated for the project to be conducted in three tasks. A summary of what is currently known about the prehistoric occupation of the region was the first task. Both literature and collections review were specifically included as sources. Updating the State Archaeologist site files for known sites and locating new sites was the second task, including both checking on sites reported by private collectors and new survey. The fourth objective was to develop a narrative predictive model of prehistoric site location for agency managers prehistoric archaeological sites (a total of 34). The field survey for new sites, most of which had to be conducted in fall 2010 before the ground froze, focused on a GIS compilation of several environmental factors previously thought to be important predictors of prehistoric site locations: distance to water (50 and 100 m) and slope (5%, 7%, 10%). Where possible, information on shoreline features from glacial lakes was also incorporated. Factors of access and land ownership (public lands preferable) were also compiled. Specific locations were then selected by the field archaeologist, drawing on decades of experience. Survey followed standard procedures, mainly shovel testing but pedestrian walkover where possible. Only four new archaeological sites were located, all in Jay Cooke State Park and all initially found by surface finds in Park trails. “Most of the areas selected for survey (County and City lands in St. Louis County, Split Rock Lighthouse State Park in Lake County, Judge Magney State Park in Cook County) were either negative or untestable. Concurrently with preparation for new survey, informant reports of new sites were recorded and, where possible, field verified in fall 2010. The response from members of the local archaeological society was overwhelming; over three dozen leads were given, about half for prehistoric sites and half for historic sites that might have prehistoric components. Additional site leads were supplied from members of the general public who heard of the project from media releases or by word of mouth. Although some site leads were either natural items or outside the Region 9 boundary, several prehistoric sites were recorded from this source. The data from both sources suggest that sites in Region 9 do tend to correlate to water, both past and present; however, specific numerical values for environmental variables are not as productive as viewing topography in the field. Areas of low slope are also high potential but not all flat area near water have site materials. Areas of low to no potential would include areas of steep slope (but not all area greater than 10% slope), water saturation (both permanent and seasonal), and/or areas at far distances from water (although quantitative measures are not firm). Much more research needs to be conducted to understand the pre-Contact occupation of the region, as well as develop a better predictive model of site location.

Gene response profiles for Daphnia pulex exposed to the environmental stressor cadmium reveals novel crustacean metallothioneins

<p>Abstract</p> <p>Background</p> <p>Genomic research tools such as microarrays are proving to be important resources to study the complex regulation of genes that respond to environmental perturbations. A first generation cDNA microarray was developed for the environmental indicator species <it>Daphnia pulex</it>, to identify genes whose regulation is modulated following exposure to the metal stressor cadmium. Our experiments revealed interesting changes in gene transcription that suggest their biological roles and their potentially toxicological features in responding to this important environmental contaminant.</p> <p>Results</p> <p>Our microarray identified genes reported in the literature to be regulated in response to cadmium exposure, suggested functional attributes for genes that share no sequence similarity to proteins in the public databases, and pointed to genes that are likely members of expanded gene families in the <it>Daphnia </it>genome. Genes identified on the microarray also were associated with cadmium induced phenotypes and population-level outcomes that we experimentally determined. A subset of genes regulated in response to cadmium exposure was independently validated using quantitative-realtime (Q-RT)-PCR. These microarray studies led to the discovery of three genes coding for the metal detoxication protein metallothionein (MT). The gene structures and predicted translated sequences of <it>D. pulex </it>MTs clearly place them in this gene family. Yet, they share little homology with previously characterized MTs.</p> <p>Conclusion</p> <p>The genomic information obtained from this study represents an important first step in characterizing microarray patterns that may be diagnostic to specific environmental contaminants and give insights into their toxicological mechanisms, while also providing a practical tool for evolutionary, ecological, and toxicological functional gene discovery studies. Advances in <it>Daphnia </it>genomics will enable the further development of this species as a model organism for the environmental sciences.</p

Examining the impact of a universal social and emotional learning intervention (Passport) on internalising symptoms and other outcomes among children, compared to the usual school curriculum: study protocol for a school-based cluster randomised trial.

Background: School-based universal social and emotional learning (SEL) interventions implemented during the transition to adolescence may be efficacious in preventing the development of mental health difficulties. This protocol describes a two-arm parallel cluster randomised controlled trial to investigate the impact of a universal SEL intervention (Passport, compared to usual provision) on internalising symptoms (primary outcome), emotion regulation, well-being, loneliness, social support, bullying, academic attainment, and health-related quality of life in English primary school pupils aged 9-11 years. A developer-led trial demonstrated the feasibility, acceptability, and utility of Passport; this will be the first independent trial.Methods: Sixty primary schools will be recruited across the Greater Manchester city region and surrounding areas, involving 2400 pupils aged 8-9 at baseline. Schools will be allocated to the intervention arm to implement Passport over 18 weekly sessions or to the control arm to implement the usual school curriculum. Random allocation will be at school level following completion of baseline measures, with minimisation to ensure balance across trial arms in school size and free school meal eligibility. Measures will be collected at baseline, post-intervention (12 months post-baseline), and at 12 months follow-up (24 months post-baseline). The primary outcome analysis (intervention effects on internalising symptoms at post-intervention) will comprise a two-level (school, child) hierarchical linear model, following the intention-to-treat principle. Additional analyses will be undertaken to assess intervention effects on secondary outcomes, maintenance effects for all outcomes, intervention compliance moderator effects, subgroup moderator effects, and mechanisms underpinning intervention effects on the primary outcome. A mixed-methods implementation and process evaluation will examine factors that influence implementation, and a health economic evaluation will assess the cost-effectiveness of the intervention.Discussion: Findings will provide educators with crucial knowledge of whether and how increasing emotion regulation through a universal intervention impacts internalising symptoms and a range of related outcomes. Findings will also inform policy related to the promotion of mental health among children and young people. If the intervention is found to be efficacious in reducing internalising symptoms and is also cost-effective, it may offer high potential as a preventative intervention for widespread implementation.Trial Registration: ISRCTN12875599; registered on 24 November 2022.</p

Arctic system on trajectory to new state

The Arctic system is moving toward a new state that falls outside the envelope of glacial-interglacial fluctuations that prevailed during recent Earth history. This future Arctic is likely to have dramatically less permanent ice than exists at present. At the present rate of change, a summer ice-free Arctic Ocean within a century is a real possibility, a state not witnessed for at least a million years. The change appears to be driven largely by feedback-enhanced global climate warming, and there seem to be few, if any processes or feedbacks within the Arctic system that are capable of altering the trajectory toward this “super interglacial” state