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IntroductionUnderstanding how abundance, productivity and distribution of individual species may respond to climate change is a critical first step towards anticipating alterations in marine ecosystem structure and function, as well as developing strategies to adapt to the full range of potential changes.MethodsThis study applies the NOAA (National Oceanic and Atmospheric Administration) Fisheries Climate Vulnerability Assessment method to 64 federally-managed species in the California Current Large Marine Ecosystem to assess their vulnerability to climate change, where vulnerability is a function of a species’ exposure to environmental change and its biological sensitivity to a set of environmental conditions, which includes components of its resiliency and adaptive capacity to respond to these new conditions.ResultsOverall, two-thirds of the species were judged to have Moderate or greater vulnerability to climate change, and only one species was anticipated to have a positive response. Species classified as Highly or Very Highly vulnerable share one or more characteristics including: 1) having complex life histories that utilize a wide range of freshwater and marine habitats; 2) having habitat specialization, particularly for areas that are likely to experience increased hypoxia; 3) having long lifespans and low population growth rates; and/or 4) being of high commercial value combined with impacts from non-climate stressors such as anthropogenic habitat degradation. Species with Low or Moderate vulnerability are either habitat generalists, occupy deep-water habitats or are highly mobile and likely to shift their ranges.DiscussionAs climate-related changes intensify, this work provides key information for both scientists and managers as they address the long-term sustainability of fisheries in the region. This information can inform near-term advice for prioritizing species-level data collection and research on climate impacts, help managers to determine when and where a precautionary approach might be warranted, in harvest or other management decisions, and help identify habitats or life history stages that might be especially effective to protect or restore.

Surface-canopy forming kelps provide the foundation for ecosystems that are ecologically, culturally, and economically important. However, these kelp forests are naturally dynamic systems that are also threatened by a range of global and local pressures. As a result, there is a need for tools that enable managers to reliably track changes in their distribution, abundance, and health in a timely manner. Remote sensing data availability has increased dramatically in recent years and this data represents a valuable tool for monitoring surface-canopy forming kelps. However, the choice of remote sensing data and analytic approach must be properly matched to management objectives and tailored to the physical and biological characteristics of the region of interest. This review identifies remote sensing datasets and analyses best suited to address different management needs and environmental settings using case studies from the west coast of North America. We highlight the importance of integrating different datasets and approaches to facilitate comparisons across regions and promote coordination of management strategies.

Major ecological realignments are already occurring in response to climate change. To be successful, conservation strategies now need to account for geographical patterns in traits sensitive to climate change, as well as climate threats to species-level diversity. As part of an effort to provide such information, we conducted a climate vulnerability assessment that included all anadromous Pacific salmon and steelhead (Oncorhynchus spp.) population units listed under the U.S. Endangered Species Act. Using an expert-based scoring system, we ranked 20 attributes for the 28 listed units and 5 additional units. Attributes captured biological sensitivity, or the strength of linkages between each listing unit and the present climate; climate exposure, or the magnitude of projected change in local environmental conditions; and adaptive capacity, or the ability to modify phenotypes to cope with new climatic conditions. Each listing unit was then assigned one of four vulnerability categories. Units ranked most vulnerable overall were Chinook (O. tshawytscha) in the California Central Valley, coho (O. kisutch) in California and southern Oregon, sockeye (O. nerka) in the Snake River Basin, and spring-run Chinook in the interior Columbia and Willamette River Basins. We identified units with similar vulnerability profiles using a hierarchical cluster analysis. Life history characteristics, especially freshwater and estuary residence times, interplayed with gradations in exposure from south to north and from coastal to interior regions to generate landscape-level patterns within each species. Nearly all listing units faced high exposures to projected increases in stream temperature, sea surface temperature, and ocean acidification, but other aspects of exposure peaked in particular regions. Anthropogenic factors, especially migration barriers, habitat degradation, and hatchery influence, have reduced the adaptive capacity of most steelhead and salmon populations. Enhancing adaptive capacity is essential to mitigate for the increasing threat of climate change. Collectively, these results provide a framework to support recovery planning that considers climate impacts on the majority of West Coast anadromous salmonids.

Networks of marine protected areas (MPAs) are being adopted globally to protect ecosystems and supplement fisheries management. The state of California recently implemented a coast-wide network of MPAs, a statewide seafloor mapping program, and ecological characterizations of species and ecosystems targeted for protection by the network. The main goals of this study were to use these data to evaluate how well seafloor features, as proxies for habitats, are represented and replicated across an MPA network and how well ecological surveys representatively sampled fish habitats inside MPAs and adjacent reference sites. Seafloor data were classified into broad substrate categories (rock and sediment) and finer scale geomorphic classifications standard to marine classification schemes using surface analyses (slope, ruggedness, etc.) done on the digital elevation model derived from multibeam bathymetry data. These classifications were then used to evaluate the representation and replication of seafloor structure within the MPAs and across the ecological surveys. Both the broad substrate categories and the finer scale geomorphic features were proportionately represented for many of the classes with deviations of 1-6% and 0-7%, respectively. Within MPAs, however, representation of seafloor features differed markedly from original estimates, with differences ranging up to 28%. Seafloor structure in the biological monitoring design had mismatches between sampling in the MPAs and their corresponding reference sites and some seafloor structure classes were missed entirely. The geomorphic variables derived from multibeam bathymetry data for these analyses are known determinants of the distribution and abundance of marine species and for coastal marine biodiversity. Thus, analyses like those performed in this study can be a valuable initial method of evaluating and predicting the conservation value of MPAs across a regional network.

Abstract High northern latitude changes with Arctic amplification across a latitudinal forest gradient suggest a shift towards an increased presence of trees and shrubs. The persistence of change may depend on the future scenarios of climate and on the current state, and site history, of forest structure. Here, we explore the persistence of a gradient-based shift in the boreal by connecting current forest patterns to recent tree cover trends and future modeled estimates of canopy height through 2100. Results show variation in the predicted potential height changes across the structural gradient from the boreal forest through the taiga-tundra ecotone. Positive potential changes in height are concentrated in transitional forests, where recent positive changes in cover prevail, while potential change in boreal forest is highly variable. Results are consistent across climate scenarios, revealing a persistent biome shift through 2100 in North America concentrated in transitional landscapes regardless of climate scenario.

T cell receptor (TCR) T cell therapies target tumor antigens in a human leukocyte antigen (HLA)-restricted manner. Biomarker-defined therapies require validation of assays suitable for determination of patient eligibility. For clinical trials evaluating TCR T cell therapies targeting melanoma-associated antigen A4 (MAGE-A4), screening in studies NCT02636855 and NCT04044768 assesses patient eligibility based on: (1) high-resolution HLA typing and (2) tumor MAGE-A4 testing via an immunohistochemical assay in HLA-eligible patients. The HLA/MAGE-A4 assays validation, biomarker data, and their relationship to covariates (demographics, cancer type, histopathology, tissue location) are reported here. HLA-A∗02 eligibility was 44.8% (2,959/6,606) in patients from 43 sites across North America and Europe. While HLA-A∗02:01 was the most frequent HLA-A∗02 allele, others (A∗02:02, A∗02:03, A∗02:06) considerably increased HLA eligibility in Hispanic, Black, and Asian populations. Overall, MAGE-A4 prevalence based on clinical trial enrollment was 26% (447/1,750) across 10 solid tumor types, and was highest in synovial sarcoma (70%) and lowest in gastric cancer (9%). The covariates were generally not associated with MAGE-A4 expression, except for patient age in ovarian cancer and histology in non-small cell lung cancer. This report shows the eligibility rate from biomarker screening for TCR T cell therapies and provides epidemiological data for future clinical development of MAGE-A4-targeted therapies.

African trypanosomes replicate within infected mammals where they are exposed to the complement system. This system centres around complement C3, which is present in a soluble form in serum but becomes covalently deposited onto the surfaces of pathogens after proteolytic cleavage to C3b. Membrane-associated C3b triggers different complement-mediated effectors which promote pathogen clearance. To counter complement-mediated clearance, African trypanosomes have a cell surface receptor, ISG65, which binds to C3b and which decreases the rate of trypanosome clearance in an infection model. However, the mechanism by which ISG65 reduces C3b function has not been determined. We reveal through cryogenic electron microscopy that ISG65 has two distinct binding sites for C3b, only one of which is available in C3 and C3d. We show that ISG65 does not block the formation of C3b or the function of the C3 convertase which catalyses the surface deposition of C3b. However, we show that ISG65 forms a specific conjugate with C3b, perhaps acting as a decoy. ISG65 also occludes the binding sites for complement receptors 2 and 3, which may disrupt recruitment of immune cells, including B cells, phagocytes, and granulocytes. This suggests that ISG65 protects trypanosomes by combining multiple approaches to dampen the complement cascade.

Lesson Study provides an opportunity for teachers to engage in layers of individual and collaborative reflections in, on, and for practice to improve the teaching and learning process. This paper reports on reflections on teaching ratio and proportion online using Lesson Study, which could improve the teaching-learning process, specifically on pedagogy. The study consisted of three major stages: Lesson Planning, Lesson Study Proper, and Reflection through post-lesson discussion. The research lesson study proper was participated in by Grade 9 students in a school in the Philippines and six teachers—one taught the lesson, and the other five observed the class via Zoom. Also, the six teachers were participants in the post-lesson discussion. The data gathered from this descriptive qualitative study were analyzed using thematic analysis. It was concluded that in teaching ratio and proportion, the distinction between terminologies must be emphasized so there will be no confusion and for the students to use what is proper. Moreover, it is advised that a real-life problem involving ratio and proportion be included in the lesson so that students will appreciate its uses and lessen its abstraction. Teachers must also go beyond routine activities to increase students' critical thinking and employ the Higher Order Thinking Skills. Thus, it is recommended that teachers undergo training to learn different and new pedagogies to improve their teaching by going beyond routine activities and incorporating real-life problems.

Abstract Background Protists of the family Trypanosomatidae (phylum Euglenozoa) have gained notoriety as parasites affecting humans, domestic animals, and agricultural plants. However, the true extent of the group's diversity spreads far beyond the medically and veterinary relevant species. We address several knowledge gaps in trypanosomatid research by undertaking sequencing, assembly, and analysis of genomes from previously overlooked representatives of this protistan group. Results We assembled genomes for twenty-one trypanosomatid species, with a primary focus on insect parasites and Trypanosoma spp. parasitizing non-human hosts. The assemblies exhibit sizes consistent with previously sequenced trypanosomatid genomes, ranging from approximately 18 Mb for Obscuromonas modryi to 35 Mb for Crithidia brevicula and Zelonia costaricensis. Despite being the smallest, the genome of O. modryi has the highest content of repetitive elements, contributing nearly half of its total size. Conversely, the highest proportion of unique DNA is found in the genomes of Wallacemonas spp., with repeats accounting for less than 8% of the assembly length. The majority of examined species exhibit varying degrees of aneuploidy, with trisomy being the most frequently observed condition after disomy. Conclusions The genome of Obscuromonas modryi represents a very unusual, if not unique, example of evolution driven by two antidromous forces: i) increasing dependence on the host leading to genomic shrinkage and ii) expansion of repeats causing genome enlargement. The observed variation in somy within and between trypanosomatid genera suggests that these flagellates are largely predisposed to aneuploidy and, apparently, exploit it to gain a fitness advantage. High heterogeneity in the genome size, repeat content, and variation in chromosome copy numbers in the newly-sequenced species highlight the remarkable genome plasticity exhibited by trypanosomatid flagellates. These new genome assemblies are a robust foundation for future research on the genetic basis of life cycle changes and adaptation to different hosts in the family Trypanosomatidae.

The Twitter interface famously prompts users to submit content by asking the question, ‘What’s happening?’. As Twitter has seen rapid change of leadership and the implications of this, emerging re-branded as X, it is a timely question to reflect back on itself, and upon the social media landscape more broadly. While the relationship between social media and higher education is far from new, the uncertainty and migration to other platforms highlight the precarity of relying on corporate infrastructure to support public scholarship. It is against this background that we launched a call for submissions to this special collection on the topic of social media in higher education. In this editorial, we will further discuss the recent changes to the social media landscape. We will focus on the JIME Twitter account as a case study attempting to make sense of the impact of this. Finally, we will introduce the papers included in the special collection, and concluding remarks.

Trypanosomes evade the immune response through antigenic variation of a surface coat containing variant surface glycoproteins (VSG). They also express invariant surface glycoproteins (ISGs), which are less well understood. Here, Macleod et al. show that ISG65 of T. brucei is a receptor for complement component 3. They provide the crystal structure of T. brucei ISG65 in complex with complement C3d and show evidence that ISG65 is involved in reducing trypanosome susceptibility to C3-mediated clearance in vivo.

Abstract African trypanosomes, such as Trypanosoma brucei, are flagellated protozoa which proliferate in mammals and cause a variety of diseases in people and animals. In a mammalian host, the external face of the African trypanosome plasma membrane is covered by a densely packed coat formed of variant surface glycoprotein (VSG), which counteracts the host's adaptive immune response by antigenic variation. The VSG is attached to the external face of the plasma membrane by covalent attachment of the C-terminus to glycosylphosphatidylinositol. As the trypanosome grows, newly synthesised VSG is added to the plasma membrane by vesicle fusion to the flagellar pocket, the sole location of exo- and endocytosis. Snake venoms contain dozens of components, including proteases and phospholipases A2. Here, we investigated the effect of Naja nigricollis venom on T. brucei with the aim of describing the response of the trypanosome to hydrolytic attack on the VSG. We found no evidence for VSG hydrolysis, however, N. nigricollis venom caused: (i) an enlargement of the flagellar pocket, (ii) the Rab11 positive endosomal compartments to adopt an abnormal dispersed localisation, and (iii) cell cycle arrest prior to cytokinesis. Our results indicate that a single protein family, the phospholipases A2 present in N. nigricollis venom, may be necessary and sufficient for the effects. This study provides new molecular insight into T. brucei biology and possibly describes mechanisms that could be exploited for T. brucei targeting.

Gene expression in pathogenic protozoans of the family Trypanosomatidae has several novel features, including multiple eIF4F-like complexes involved in protein synthesis. The eukaryotic eIF4F complex, formed mainly by eIF4E and eIF4G subunits, is responsible for the canonical selection of mRNAs required for the initiation of mRNA translation. The best-known complexes implicated in translation in trypanosomatids are based on two related pairs of eIF4E and eIF4G subunits (EIF4E3/EIF4G4 and EIF4E4/EIF4G3), whose functional distinctions remain to be fully described. Here, to define interactomes associated with both complexes in Trypanosoma brucei procyclic forms, we performed parallel immunoprecipitation experiments followed by identification of proteins co-precipitated with the four tagged eIF4E and eIF4G subunits. A number of different protein partners, including RNA binding proteins and helicases, specifically co-precipitate with each complex. Highlights with the EIF4E4/EIF4G3 pair include RBP23, PABP1, EIF4AI and the CRK1 kinase. Co-precipitated partners with the EIF4E3/EIF4G4 pair are more diverse and include DRBD2, PABP2 and different zinc-finger proteins and RNA helicases. EIF4E3/EIF4G4 are essential for viability and to better define their role, we further investigated their phenotypes after knockdown. Depletion of either EIF4E3/EIF4G4 mRNAs lead to aberrant morphology with a more direct impact on events associated with cytokinesis. We also sought to identify those mRNAs differentially associated with each complex through CLIP-seq with the two eIF4E subunits. Predominant among EIF4E4-bound transcripts are those encoding ribosomal proteins, absent from those found with EIF4E3, which are generally more diverse. RNAi mediated depletion of EIF4E4, which does not affect proliferation, does not lead to changes in mRNAs or proteins associated with EIF4E3, confirming a lack of redundancy and distinct roles for the two complexes.

Purpose: The sociopolitical determinants of health drive health outcomes and inequities in the United States. Primary care practices are, increasingly, expected by payers and policy makers to assess patients’ social needs. Resource referral platforms provide physicians with information and referral systems for community resources. One commonly used platform is Aunt Bertha/Find Help (AB/FH). The American Academy of Family Physicians (AAFP) Neighborhood Navigator (NN) tool allows physicians and laypeople to search for resources using AB/FH. We sought to describe what users were searching for and to identify patterns to inform resource allocation. Methods: This was a descriptive study of the AAFP’s NN tool. Searches of NN were analyzed to describe what users were searching for. Results: From 2018 to April 2022 there were 168 135 searches. The most common searches were for food and housing insecurity (22%, 21%) and health care referral (20.6%) with 22% more searches in the winter than the spring. There was a 119% increase in searches between 2018 and 2022, and a 47% increase in searches during the COVID-19 Pandemic. In the “Health” category the top 20 subcategories accounted for over 77% of searches. Conclusions: Family physicians and their patients use NN to search AB/FH for community resources to address adverse social determinants of health (SDOH). As expected, searches increased during the COVID-19 pandemic. This type of analysis may help individual clinicians, practices, and health systems prepare for the most common social needs of their patients. Social resource platforms might serve as a robust measure for primary care practice screening and referral for SDOH.

Lori Peacock et al. report the sequence of meiosis in Trypanosoma brucei cells from the salivary glands of tsetse flies using image analyses and expression of the cell fusion protein HAP2. From the recovered cell types and expression of HAP2 in meiotic intermediates, these results indicate that haploid gametes are produced sequentially.

BackgroundAnimal African Trypanosomiasis (AAT) is one of the most economically important diseases affecting livestock productivity in sub-Saharan Africa. The disease is caused by a broad range of Trypanosoma spp., infecting both wild and domesticated animals through cyclical and mechanical transmission. This study aimed to characterize trypanosomes present in cattle at regular intervals over two years in an AAT endemic and a non-endemic region of Ghana.Methodology/principal findingsGroups of cattle at Accra and Adidome were selected based on their geographical location, tsetse fly density, prevalence of trypanosomiasis and the breed of cattle available. Blood for DNA extraction was collected at approximately four to five-week intervals over a two-year period. Trypanosome DNA were detected by a sensitive nested PCR targeting the tubulin gene array and massively parallel sequencing of barcoded amplicons. Analysis of the data was a semi-quantitative estimation of infection levels using read counts obtained from the sequencing as a proxy for infection levels. Majority of the cattle were infected with multiple species most of the time [190/259 (73%) at Adidome and 191/324 (59%) at Accra], with T. vivax being the most abundant. The level of infection and in particular T. vivax, was higher in Adidome, the location with a high density of tsetse flies. The infection level varied over the time course, the timings of this variation were not consistent and in Adidome it appeared to be independent of prophylactic treatment for trypanosome infection. Effect of gender or breed on infection levels was insignificant.Conclusions/significanceMost cattle were infected with low levels of several trypanosome species at both study sites, with T. vivax being the most abundant. The measurements of infection over time provided insight to the importance of the approach in identifying cattle that could suppress trypanosome infection over an extended time and may serve as reservoir.

African trypanosome infections can persist for years, but immune evasion mechanisms are not fully understood. Here, Macleod et al. identify a trypanosome receptor for mammalian factor H, a negative regulator of the alternative complement pathway, that increases parasite transmission to tsetse flies.

Anaplasmosis, caused by infection with bacteria of the genus Anaplasma, is an important veterinary and zoonotic disease. Transmission by ticks has been characterized but little is known about non-tick vectors of livestock anaplasmosis. This study investigated the presence of Anaplasma spp. in camels in northern Kenya and whether the hematophagous camel ked, Hippobosca camelina, acts as a vector. Camels (n = 976) and > 10,000 keds were sampled over a three-year study period and the presence of Anaplasma species was determined by PCR-based assays targeting the Anaplasmataceae 16S rRNA gene. Camels were infected by a single species of Anaplasma, 'Candidatus Anaplasma camelii', with infection rates ranging from 63-78% during the dry (September 2017), wet (June-July 2018), and late wet seasons (July-August 2019). 10-29% of camel keds harbored 'Ca. Anaplasma camelii' acquired from infected camels during blood feeding. We determined that Anaplasma-positive camel keds could transmit 'Ca. Anaplasma camelii' to mice and rabbits via blood-feeding. We show competence in pathogen transmission and subsequent infection in mice and rabbits by microscopic observation in blood smears and by PCR. Transmission of 'Ca. Anaplasma camelii' to mice (8-47%) and rabbits (25%) occurred readily after ked bites. Hence, we demonstrate, for the first time, the potential of H. camelina as a vector of anaplasmosis. This key finding provides the rationale for establishing ked control programmes for improvement of livestock and human health.

Background: The left ventricle is built to carry out its function as a powerful pump. To detail recent findings of anatomical variants associated with the left ventricular compartment, we will first review the external and general features, followed by the internal features and subsequently, the standard variants that can exist within this chamber of the heart. Methods: This literature review seeks to collate and discuss peer-reviewed articles on the anatomy of the left ventricle. Results: The left ventricle has many unique features including walls that are thicker than those of the right ventricle, an overlap of its inlet and outlet portions, and the hinge of the leaflets of the mitral valve that are oriented cranially relative to those of the tricuspid valve. Conclusion: While the left ventricle shares many characteristics with that of the right ventricle, the unique features may contribute to cardiac pathology and can also be used to the surgeon's advantage in treating disease.

Lacanienta and his colleagues (2018) recently reported results of a study of the effect of themes on quality of experience of youths during summer camp activities. Existing literature suggested that theming activity sessions would have a strong main effect. Results, though, revealed an activity-by-theme interaction effect, i.e., themes seem to be effective in some activities but not others. In this follow-up study, we describe results of a secondary analysis revealing significant new insights regarding theme. Adding an indicator of campers’ co-created, lived-experience theme into the models tested substantially clarified how objective theme, lived-experience theme, and activity interact in influencing the quality of structured experiences. This study, then, underscores the importance of including measures of participants’ co-created lived experience as we seek to understand techniques that can be used to enhance the quality of youths’ structured camp experiences.

Abstract Background Photosynthetic euglenids are major contributors to fresh water ecosystems. Euglena gracilis in particular has noted metabolic flexibility, reflected by an ability to thrive in a range of harsh environments. E. gracilis has been a popular model organism and of considerable biotechnological interest, but the absence of a gene catalogue has hampered both basic research and translational efforts. Results We report a detailed transcriptome and partial genome for E. gracilis Z1. The nuclear genome is estimated to be around 500 Mb in size, and the transcriptome encodes over 36,000 proteins and the genome possesses less than 1% coding sequence. Annotation of coding sequences indicates a highly sophisticated endomembrane system, RNA processing mechanisms and nuclear genome contributions from several photosynthetic lineages. Multiple gene families, including likely signal transduction components, have been massively expanded. Alterations in protein abundance are controlled post-transcriptionally between light and dark conditions, surprisingly similar to trypanosomatids. Conclusions Our data provide evidence that a range of photosynthetic eukaryotes contributed to the Euglena nuclear genome, evidence in support of the ‘shopping bag’ hypothesis for plastid acquisition. We also suggest that euglenids possess unique regulatory mechanisms for achieving extreme adaptability, through mechanisms of paralog expansion and gene acquisition.

Mutations in the Trypanosoma brucei aquaporin AQP2 are associated with resistance to pentamidine and melarsoprol. We show that TbAQP2 but not TbAQP3 was positively selected for increased pore size from a common ancestor aquaporin. We demonstrate that TbAQP2’s unique architecture permits pentamidine permeation through its central pore and show how specific mutations in highly conserved motifs affect drug permeation. Introduction of key TbAQP2 amino acids into TbAQP3 renders the latter permeable to pentamidine. Molecular dynamics demonstrates that permeation by dicationic pentamidine is energetically favourable in TbAQP2, driven by the membrane potential, although aquaporins are normally strictly impermeable for ionic species. We also identify the structural determinants that make pentamidine a permeant although most other diamidine drugs are excluded. Our results have wide-ranging implications for optimising antitrypanosomal drugs and averting cross-resistance. Moreover, these new insights in aquaporin permeation may allow the pharmacological exploitation of other members of this ubiquitous gene family.

Background: Major constraints to camel production include pests and diseases. In northern Kenya, little information is available about blood-borne pathogens circulating in one-humped camels ( Camelus dromedarius) or their possible transmission by the camel haematophagous ectoparasite, Hippobosca camelina, commonly known as camel ked or camel fly. This study aimed to: (i) identify the presence of potentially insect-vectored pathogens in camels and camel keds, and (ii) assess the potential utility of keds for xenodiagnosis of camel pathogens that they may not vector. Methods: In Laisamis, northern Kenya, camel blood samples (n = 249) and camel keds (n = 117) were randomly collected from camels. All samples were screened for trypanosomal and camelpox DNA by PCR, and for Anaplasma, Ehrlichia, Brucella, Coxiella, Theileria, and Babesia by PCR coupled with high-resolution melting (PCR-HRM) analysis. Results: In camels, we detected Trypanosoma vivax (41%), Trypanosoma evansi (1.2%), and “ Candidatus Anaplasma camelii” (68.67%). In camel keds, we also detected T. vivax (45.3%), T. evansi (2.56%), Trypanosoma melophagium (1/117) (0.4%), and “ Candidatus Anaplasma camelii” (16.24 %). Piroplasms ( Theileria spp. and Babesia spp.), Coxiella burnetii, Brucella spp., Ehrlichia spp., and camel pox were not detected in any samples. Conclusions: This study reveals the presence of epizootic pathogens in camels from northern Kenya. Furthermore, the presence of the same pathogens in camels and in keds collected from sampled camels suggests the potential use of these flies in xenodiagnosis of haemopathogens circulating in camels.

Pollen nutrition is necessary for proper growth and development of adult honey bees. Yet, it is unclear how pollen affects the honey bee brain and behavior. We investigated whether pollen affects amino acids in the brains of caged, nurse-aged bees, and what the behavioral consequences might be. We also tested whether parasitic stress altered this relationship by analyzing bees infected with prevalent stressor, Nosema ceranae. Levels of 18 amino acids in individual honey bee brains were measured using Gas Chromatography – Mass Spectrometry at two different ages (Day 7 and Day 11). We then employed the proboscis extension reflex to test odor learning and memory. We found that the honey bee brain was highly responsive to pollen. Many amino acids in the brain were elevated and were present at higher concentration with age. The majority of these amino acids were non-essential. Without pollen, levels of amino acids remained consistent, or declined. Nosema-infected bees showed a different profile. Infection altered amino acid levels in a pollen-dependent manner. The majority of amino acids were lower when pollen was given, but higher when pollen was deprived. Odor learning and memory was not affected by feeding pollen to uninfected bees; but pollen did improve performance in Nosema-infected bees. These results suggest that pollen in early adulthood continues to shape amino acid levels in the brain with age, which may affect neural circuitry and behavior over time. Parasitic stress by N. ceranae modifies this relationship revealing an interaction between infection, pollen nutrition, and behavior.

Summary: Ribosome assembly occurs mainly in the nucleolus, yet recent studies have revealed robust enrichment and translation of mRNAs encoding many ribosomal proteins (RPs) in axons, far away from neuronal cell bodies. Here, we report a physical and functional interaction between locally synthesized RPs and ribosomes in the axon. We show that axonal RP translation is regulated through a sequence motif, CUIC, that forms an RNA-loop structure in the region immediately upstream of the initiation codon. Using imaging and subcellular proteomics techniques, we show that RPs synthesized in axons join axonal ribosomes in a nucleolus-independent fashion. Inhibition of axonal CUIC-regulated RP translation decreases local translation activity and reduces axon branching in the developing brain, revealing the physiological relevance of axonal RP synthesis in vivo. These results suggest that axonal translation supplies cytoplasmic RPs to maintain/modify local ribosomal function far from the nucleolus in neurons. : Local protein synthesis in axons supplies new ribosomal proteins far from the nucleolus, the known site of ribosome biogenesis. Shigeoka et al. provide evidence that axonally synthesized ribosomal proteins join pre-existing ribosomes and maintain translation activity in axons, which is required for axon terminal branching. Keywords: axon, mRNA, local translation, ribosome, ribosomal proteins, Rps4x, axonal protein synthesis, ribosome remodeling, axon branching, neural wiring

Background: The right ventricle is the most anteriorly positioned chamber of the heart, sitting directly posterior to the sternum. The distinct anatomical features of the right ventricle create an approximately 10-fold difference in vascular resistance between the right and left ventricular systems. We will first review the external and general features, followed by the internal features and subsequently, the clinical relevance of this chamber of the heart. Methods: This literature review seeks to collate and discuss peer-reviewed articles on the anatomy of the right ventricle. It was outlined after conducting a review of the literature and standard search engines. Results: Factors such as retrosternal location, complex geometry, and normal variants create complexities in accurate diagnosis of right ventricle pathology. Conclusion: Knowledge of the underlying anatomy of the right ventricle and the subsequent functional properties are examined in this review to promote improved diagnostic protocols and clinical interventions in the future. Keywords: Heart, Right ventricle, Ventricular function, Tricuspid

Using themes (such as the Wild West or Survivor or Star Wars) in camp settings may enhance the quality of camp activities and youths’ overall camp experience. We evaluated the effect of theming camp experiences on the quality of subjective experiences of campers. Campers (N = 231) in 3 sessions of a residential 4-H camp participated in the study. One camp session was fully themed (all activities used tangible and intangible props, cues, and imaginary story contexts), a second was partially themed (intangible cues and stories only), and the third was not themed. Questionnaires measuring the quality of immediate subjective experiences (N=1,847) were completed following each of 8 activity sessions (e.g., climbing, fishing, swimming). Campers also completed a questionnaire for the purpose of overall camp evaluation at the end of their camp sessions. Activity-level data were analyzed using linear mixed modeling techniques. Ordinary least-squares regression was used to analyze campers’ overall camp experiences. Results at the activity level revealed significant theme-by-activity interaction effects. At the camp level, a hypothesized causal sequence linking theme to likelihood to recommend was supported.

Using intracellular cytokine staining we show herein that T cells will respond to short-term (6 h) activation with phorbol ester plus ionomycin by production of tumor necrosis factor (TNF), IFN-γ or both. Here CD4 T cells preferentially produce TNF and CD8 cells IFN-γ. The same pattern is seen when T cells are activated with the Mycobacterium tuberculosis protein early secretory antigenic target-6 (ESAT-6). Responses with >0.02% IFN-γ+ CD3 cells were seen in 8 of 10 patients diagnosed with tuberculosis and in 12 of 14 healthy individuals selected for likely exposure to M. tuberculosis. T cell responses to the 1:1 complex of ESAT-6 and culture filtrate protein-10 (CFP-10) were inferior to ESAT-6 alone, and only reached the level of T cell response achieved with CFP-10 alone. Extending the time of incubation to 18 h leads to an increased response to the complex, but it still reached only the level of CFP-10 alone. In vitro digestion with lysosomal enzymes cathepsin L and S at 2000:1 protein to enzyme ratio demonstrates rapid digestion of the individual proteins while the ESAT-6–CFP-10 complex is resistant. The data suggest that the natural complex of ESAT-6–CFP-10 is less amenable to antigen processing leading to a lower T cell response as compared with the individual proteins. [ABSTRACT FROM AUTHOR]

Abstract Background Animal African trypanosomiasis (AAT) greatly affects livestock production in sub-Saharan Africa. In Ghana prevalence of AAT is estimated to range between 5 and 50%. Studies have reported serum biochemical aberrations and variability in cytokine profiles in animals during infection. However, information regarding the biochemical parameters and cytokine profiles associated with natural infections are limited. This study was therefore aimed at investigating changes in the levels of serum biochemical parameters and inflammatory cytokines during a natural infection. Methods Nested internal transcribed spacer (ITS)-based PCR and sequencing were used to characterise trypanosome infection in cattle at two areas in Ghana (Adidome and Accra) of different endemicities. The cattle were sampled at four to five-week intervals over a period of six months. Levels of serum biochemical parameters, including creatinine, cholesterol, alkaline phosphatase (ALP), alanine aminotransferase (ALT), total bilirubin and total protein and cytokines (interleukin 10, interleukin 4, interleukin 12, interferon gamma and tumor necrosis factor alpha) were measured in serum samples and then compared between infected cattle and uninfected controls. Results The predominant trypanosome species detected in Accra (non-endemic) and Adidome (endemic) were Trypanosoma theileri and Trypanosoma vivax, respectively. Serum biochemical parameters were similar between infected and uninfected cattle in Accra. Infected cattle at Adidome however, had significantly higher levels of ALP, creatinine, total protein and total bilirubin (P

Abstract Background In changing higher education environments, medical educators are increasingly challenged to prepare new doctors to care for ageing populations. The Depth of Field: Exploring Ageing resource (DOF) uses photographs, reflective questioning prompts, older adults’ narratives and collaborative dialogue to foster anticipatory reflection or ‘preflection’ in medical students prior to their first geriatric medicine clinical placement. The aim of this research is to explore whether photographs, narratives and small group collaborative dialogue fosters reflective learning, enhances reflective capacity and has the potential to shift medical students’ attitudes towards caring for older adults. Methods This study used a mixed method evaluation design, measuring attitudes using pre and post questionnaire responses and individual written reflections drawn from 128 second year medical students, exploring their perceptions toward older adults. Results Quantitative and qualitative data indicated that the DOF session generated reflective learning that resulted in positive shifts in medical students’ perceptions towards older adults. The qualitative reflections were captured in four main themes: the opportunity provided to Envision working with older adults; the Tension created to challenge learners’ misinformed assumptions, and the work of Dismantling those assumptions, leading to Seeing older people as individuals. Conclusions These findings highlight how visual and narrative methodologies can be used as an effective reflective learning tool to challenge medical students’ assumptions around ageing and how these may influence their care of older adults.

Trypanosomes are important disease agents of humans, livestock and cold-blooded species, including fish. The cellular morphology of trypanosomes is central to their motility, adaptation to the host’s environments and pathogenesis. However, visualizing the behaviour of trypanosomes resident in a live vertebrate host has remained unexplored. In this study, we describe an infection model of zebrafish (Danio rerio) with Trypanosoma carassii. By combining high spatio-temporal resolution microscopy with the transparency of live zebrafish, we describe in detail the swimming behaviour of trypanosomes in blood and tissues of a vertebrate host. Besides the conventional tumbling and directional swimming, T. carassii can change direction through a ‘whip-like’ motion or by swimming backward. Further, the posterior end can act as an anchoring site in vivo. To our knowledge, this is the first report of a vertebrate infection model that allows detailed imaging of trypanosome swimming behaviour in vivo in a natural host environment.

Infections of humans and livestock with African trypanosomes are treated with drugs introduced decades ago that are not always fully effective and often have severe side effects. Here, the trypanosome haptoglobin-haemoglobin receptor (HpHbR) has been exploited as a route of uptake for an antibody-drug conjugate (ADC) that is completely effective against Trypanosoma brucei in the standard mouse model of infection. Recombinant human anti-HpHbR monoclonal antibodies were isolated and shown to be internalised in a receptor-dependent manner. Antibodies were conjugated to a pyrrolobenzodiazepine (PBD) toxin and killed T. brucei in vitro at picomolar concentrations. A single therapeutic dose (0.25 mg/kg) of a HpHbR antibody-PBD conjugate completely cured a T. brucei mouse infection within 2 days with no re-emergence of infection over a subsequent time course of 77 days. These experiments provide a demonstration of how ADCs can be exploited to treat protozoal diseases that desperately require new therapeutics.

Honey bee colonies have a yearly cycle that is supported nutritionally by the seasonal progression of flowering plants. In the spring, colonies grow by rearing brood, but in the fall, brood rearing declines in preparation for overwintering. Depending on where colonies are located, the yearly cycle can differ especially in overwintering activities. In temperate climates of Europe and North America, colonies reduce or end brood rearing in the fall while in warmer climates bees can rear brood and forage throughout the year. To test the hypothesis that nutrients available in seasonal pollens and honey bee responses to them can differ we analyzed pollen in the spring and fall collected by colonies in environments where brood rearing either stops in the fall (Iowa) or continues through the winter (Arizona). We fed both types of pollen to worker offspring of queens that emerged and open mated in each type of environment. We measured physiological responses to test if they differed depending on the location and season when the pollen was collected and the queen line of the workers that consumed it. Specifically, we measured pollen and protein consumption, gene expression levels (hex 70, hex 110, and vg) and hypopharyngeal gland (HPG) development. We found differences in macronutrient content and amino and fatty acids between spring and fall pollens from the same location and differences in nutrient content between locations during the same season. We also detected queen type and seasonal effects in HPG size and differences in gene expression between bees consuming spring vs. fall pollen with larger HPG and higher gene expression levels in those consuming spring pollen. The effects might have emerged from the seasonal differences in nutritional content of the pollens and genetic factors associated with the queen lines we used.

The hyper-virulent emm59 genotype of invasive group A Streptococcus was identified in northern Arizona in 2015. Eighteen isolates belonging to a genomic cluster grouped most closely with recently identified isolates in New Mexico. The continued transmission of emm59 in the southwestern United States poses a public health concern.

Lack of national data on water-related ecosystems is a major challenge to achieving the Sustainable Development Goal (SDG) 6 targets by 2030. Monitoring surface water extent, wetlands, and water quality from space can be an important asset for many countries in support of SDG 6 reporting. We demonstrate the potential for Earth observation (EO) data to support country reporting for SDG Indicator 6.6.1, ‘Change in the extent of water-related ecosystems over time’ and identify important considerations for countries using these data for SDG reporting. The spatial extent of water-related ecosystems, and the partial quality of water within these ecosystems is investigated for seven countries. Data from the Moderate Resolution Imaging Spectroradiometer (MODIS) and Landsat 5, 7, and 8 with Shuttle Radar Topography Mission (SRTM) are used to measure surface water extent at 250 m and 30 m spatial resolution, respectively, in Cambodia, Jamaica, Peru, the Philippines, Senegal, Uganda, and Zambia. The extent of mangroves is mapped at 30 m spatial resolution using Landsat 8 Operational Land Imager (OLI), Sentinel-1, and SRTM data for Jamaica, Peru, and Senegal. Using Landsat 8 and Sentinel 2A imagery, total suspended solids and chlorophyll-a are mapped over time for a select number of large surface water bodies in Peru, Senegal, and Zambia. All of the EO datasets used are of global coverage and publicly available at no cost. The temporal consistency and long time-series of many of the datasets enable replicability over time, making reporting of change from baseline values consistent and systematic. We find that statistical comparisons between different surface water data products can help provide some degree of confidence for countries during their validation process and highlight the need for accuracy assessments when using EO-based land change data for SDG reporting. We also raise concern that EO data in the context of SDG Indicator 6.6.1 reporting may be more challenging for some countries, such as small island nations, than others to use in assessing the extent of water-related ecosystems due to scale limitations and climate variability. Country-driven validation of the EO data products remains a priority to ensure successful data integration in support of SDG Indicator 6.6.1 reporting. Multi-country studies such as this one can be valuable tools for helping to guide the evolution of SDG monitoring methodologies and provide a useful resource for countries reporting on water-related ecosystems. The EO data analyses and statistical methods used in this study can be easily replicated for country-driven validation of EO data products in the future.

Climate and weather data such as precipitation derived from Global Climate Models (GCMs) and satellite observations are essential for the global and local hydrological assessment. However, most climatic popular precipitation products (with spatial resolutions coarser than 10km) are too coarse for local impact studies and require “downscaling” to obtain higher resolutions. Traditional precipitation downscaling methods such as statistical and dynamic downscaling require an input of additional meteorological variables, and very few are applicable for downscaling hourly precipitation for higher spatial resolution. Based on dynamic dictionary learning, we propose a new downscaling method, PreciPatch, to address this challenge by producing spatially distributed higher resolution precipitation fields with only precipitation input from GCMs at hourly temporal resolution and a large geographical extent. Using aggregated Integrated Multi-satellitE Retrievals for GPM (IMERG) data, an experiment was conducted to evaluate the performance of PreciPatch, in comparison with bicubic interpolation using RainFARM—a stochastic downscaling method, and DeepSD—a Super-Resolution Convolutional Neural Network (SRCNN) based downscaling method. PreciPatch demonstrates better performance than other methods for downscaling short-duration precipitation events (used historical data from 2014 to 2017 as the training set to estimate high-resolution hourly events in 2018).

Selective transcription of individual protein coding genes does not occur in trypanosomes and the cellular copy number of each mRNA must be determined post-transcriptionally. Here, we provide evidence that codon choice directs the levels of constitutively expressed mRNAs. First, a novel codon usage metric, the gene expression codon adaptation index (geCAI), was developed that maximised the relationship between codon choice and the measured abundance for a transcriptome. Second, geCAI predictions of mRNA levels were tested using differently coded GFP transgenes and were successful over a 25-fold range, similar to the variation in endogenous mRNAs. Third, translation was necessary for the accelerated mRNA turnover resulting from codon choice. Thus, in trypanosomes, the information determining the levels of most mRNAs resides in the open reading frame and translation is required to access this information.

The degree of conservation and evolution of cytoplasmic mRNA metabolism pathways across the eukaryotes remains incompletely resolved. In this study, we describe a comprehensive genome and transcriptome-wide analysis of proteins involved in mRNA maturation, translation, and mRNA decay across representative organisms from the six eukaryotic super-groups. We demonstrate that eukaryotes share common pathways for mRNA metabolism that were almost certainly present in the last eukaryotic common ancestor, and show for the first time a correlation between intron density and a selective absence of some Exon Junction Complex (EJC) components in eukaryotes. In addition, we identify pathways that have diversified in individual lineages, with a specific focus on the unique gene gains and losses in members of the Excavata and SAR groups that contribute to their unique gene expression pathways compared to other organisms.

African trypanosomes have complex life cycles comprising at least ten developmental forms, variously adapted to different niches in their tsetse fly vector and their mammalian hosts. Unlike many other protozoan pathogens, they are always extracellular and have evolved intricate surface coats that allow them to obtain nutrients while also protecting them from the immune defenses of either insects or mammals. The acquisition of macromolecular nutrients requires receptors that function within the context of these surface coats. The best understood of these is the haptoglobin-hemoglobin receptor (HpHbR) of Trypanosoma brucei, which is used by the mammalian bloodstream form of the parasite, allowing heme acquisition. However, in some primates it also provides an uptake route for trypanolytic factor-1, a mediator of innate immunity against trypanosome infection. Recent studies have shown that during the evolution of African trypanosome species the receptor has diversified in function from a hemoglobin receptor predominantly expressed in the tsetse fly to a haptoglobin-hemoglobin receptor predominantly expressed in the mammalian bloodstream. Structural and functional studies of homologous receptors from different trypanosome species have allowed us to propose an evolutionary history for how one receptor has adapted to different roles in different trypanosome species. They also highlight the challenges that a receptor faces in operating on the complex trypanosome surface and show how these challenges can be met.

The fate of an mRNA is determined by its interaction with proteins and small RNAs within dynamic complexes called ribonucleoprotein complexes (mRNPs). In Trypanosoma brucei and related kinetoplastids, responses to internal and external signals are mainly mediated by post-transcriptional processes. Here, we used proximity-dependent biotin identification (BioID) combined with RNA-seq to investigate the changes resulting from ectopic expression of RBP10 and RBP9, two developmentally regulated RNA-binding proteins (RBPs). Both RBPs have reduced expression in insect procyclic forms (PCFs) compared with bloodstream forms (BSFs). Upon overexpression in PCFs, both proteins were recruited to cytoplasmic foci, co-localizing with the processing body marker SCD6. Further, both RBPs altered the transcriptome from a PCF- to a BSF-like pattern. Notably, upon expression of BirA*-RBP9 and BirA*-RBP10, BioID yielded more than 200 high confidence protein interactors (more than 10-fold enriched); 45 (RBP9) and 31 (RBP10) were directly related to mRNA metabolism. This study validates the use of BioID for investigating mRNP components but also illustrates the complexity of mRNP function.

Recent profound changes have been observed in the Arctic environment, including record low sea ice extents and high latitude greening. Studying the Arctic and how it is changing is an important element of climate change science. The Tundra, an ecoregion of the Arctic, is directly related to climate change due to its effects on the snow ice feedback mechanism and greenhouse gas cycling. Like all ecoregions, the Tundra border is shifting, yet studies and policies require clear delineation of boundaries. There are many options for ecoregion classification systems, as well as resources for creating custom maps. To help decision makers identify the best classification system possible, we present a review of North American Tundra ecoregion delineations and further explore the methodologies, purposes, limitations, and physical properties of five common ecoregion classification systems. We quantitatively compare the corresponding maps by area using a geographic information system.