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The Amblyomma marmoreum complex includes afrotropical species, such as Amblyomma sparsum, a three-host tick that parasitizes reptiles, birds, and mammals, and is a recognized vector of Ehrlichia ruminantium. However, the lack of morphological, genetic and ecological data on A. sparsum has caused considerable confusion in its identification. In this study, we used microscopy and metagenomic approaches to analyze A. sparsum ticks collected from a puff adder snake (Bitis arietans) in southwest Senegal (an endemic rickettsioses area) in order to supplement previous morphological descriptions, provide novel genomic data for the A. marmoreum complex, and describe the genome of a novel spotted fever group Rickettsia strain. Based on stereoscope and scanning electron microscopy (SEM) morphological evaluations, we provide high-quality images and new insights about punctation and enameling in the adult male of A. sparsum to facilitate identification for future studies. The metagenomic approach allowed us assembly the complete mitochondrial genome of A. sparsum, as well as the nearly entire chromosome and complete plasmid sequences of a novel Rickettsia africae strain. Phylogenomic analyses demonstrated a close relationship between A. sparsum and Amblyomma nuttalli for the first time and confirmed the position of A. sparsum within the A. marmoreum complex. Our results provide new insights into the systematics of A. sparsum and A. marmoreum complex, as well as the genetic diversity of R. africae in the Afrotropical region. Future studies should consider the possibility that A. sparsum may be a vector for R. africae.

Abstract Background Anxiety disorders are prevalent and disabling conditions involving excessive worry and tension. Generalized anxiety disorder (GAD), the most common anxiety disorder, affects 5% of individuals from high-income countries and many individuals report that treatment options are not accessible, effective, or tolerable. Clinical evidence suggests that nutrition interventions, based on the Mediterranean diet and supplementation of omega-3 fatty acids, can significantly improve symptoms of depression; however, the effect of nutrition interventions on anxiety symptoms has not been studied in a clinical population. The primary objective of the present study is to assess the feasibility and acceptability of a dietary counselling and omega-3 fatty acid supplementation intervention delivered to adult women with GAD. The secondary objectives include assessing changes in anxiety symptom severity, assessing changes in quality of life, assessing changes in biomarkers, and evaluating the components of the program. Methods This study is a randomized, wait-list controlled pilot trial delivering a 12-week, dietary counselling intervention and omega-3 supplementation to 50 adult women with GAD. Participants will complete seven individual counselling sessions which include education, personalized recommendations, mindful eating techniques, motivational interviewing, and goal setting. They will be provided with recipes, instructional videos, and food items. The intervention is designed based on the Social Cognitive Theory and previous research that has been done by the author team to identify dietary constituents with the most evidence to support their use in the treatment of anxiety disorders. Questionnaires and blood work will be completed at baseline, after the waiting period (for those in the waitlist group), and after the intervention. Discussion Results from this study will lay the foundation for future large-scale studies in this area and may provide preliminary evidence of the role of diet counselling and omega-3 supplementation in the management of GAD. Research on the role of nutrition in psychiatric care has been identified as a priority by a number of international organizations. The present trial directly addresses the call for the research that is most needed to advance the field. Trial registration This protocol was registered at ClinicalTrials.gov on October 10, 2022; NCT05573672 . Trial sponsor: The Canadian College of Naturopathic Medicine, 1255 Sheppard Ave E, Toronto, ON M2K 1E2, 416–498-1255. Steering committee: Composed of MA, LL, KC, IvdW, SM, UN, AJ. The committee meets monthly to oversee the trial. Protocol identifier: CCNM_EASe-GADCT_2201v4.

The tick-borne encephalitis virus (TBEV) is transmitted to humans through tick bites. In recent years, the appearance of the Siberian subtype of TBEV in Ixodes ricinus in Finland, together with deaths from the normally mild European subtype in the same country, have raised concerns about a possible spread of virulent variants of TBEV in Western Europe. Thus, there is a need to monitor the spread of strains, particularly of the European and Siberian subtypes. In this study, we develop a new real-time PCR method targeting Siberian and European subtypes of TBEV. The primers amplify a 176 bp fragment of the E gene, which is suitable for subsequent strain identification by Sanger sequencing. This study pioneers a new approach to primer design where the melting temperature (Tm) of primers annealed to representative mismatched target sequences is empirically determined and used to guide improvements in primer sequence. This allowed the range of TBEV strains detected to be extended to cover most European and Siberian strains tested, in addition to a strain of the Far-Eastern subtype. The limit of detection was 10–100 DNA copies per reaction and amplification efficiency varied between 83% and 94%, depending on the TBEV strain. Experimental determination of primer Tm proved to be a fruitful approach and will be a useful tool for future primer design and diagnostics.

Patients with myeloproliferative neoplasms (MPNs) are at increased risk of cardiovascular disease (CVD), including acute coronary syndrome (ACS). However, data on long-term outcomes of patients with MPN who have had ACS and risk factors for all-cause death or CV events post-ACS hospitalization are lacking. We conducted a single-center study of 41 consecutive patients with MPN with ACS hospitalization after MPN diagnosis. After a median follow-up of 80 months after ACS hospitalization, 31 (76%) experienced death or a CV event (myocardial infarction, ischemic stroke, or heart failure hospitalization). After multivariable Cox proportional hazards regression, index ACS within 12 months of MPN diagnosis (HR 3.84, 95% CI 1.44–10.19), WBC ≥ 20 K/µL (HR 9.10, 95% CI 2.71–30.52), JAK2 mutation (HR 3.71, 95% CI 1.22–11.22), and prior CVD (HR 2.60, 95% CI 1.12–6.08) were associated with increased death or CV events. Further studies are warranted to improve cardiovascular outcomes in this patient population.

Abstract Environmental DNA (eDNA) is an increasingly used noninvasive and cost‐effective method for species detections in surveillance studies. The myxozoan endoparasite Tetracapsuloides bryosalmonae is the causative agent of proliferative kidney disease (PKD) in salmonid fish. PKD is a potentially lethal disease of freshwater salmonids when water temperatures exceed 12–14°C for prolonged periods. Periodically, high mortality and decline in farmed and wild salmonid populations in Europe and North America have been reported in the last decades. The aim of this study was to use eDNA as a method to detect Tetracapsuloides bryosalmonae from large, deep, dimictic Norwegian lakes. Such habitats are expected to become increasingly important for cold‐water salmonids with global warming. Samples were collected from five lakes in southeastern Norway, and parasite DNA was detected by qPCR. eDNA from T. bryosalmonae was detected in four of the five lakes surveyed. These findings corresponded with the detection of T. bryosalmonae DNA in salmonid kidneys in four of the lakes in a previous survey. The detection of parasites from eDNA varied between years and stations within the same lake, revealing a changing and apparently stochastic spatial distribution of parasite DNA from year to year. Nonetheless, by sampling multiple sites throughout the lakes, we were able to detect T. bryosalmonae at the lake level in both survey years. Strategies for eDNA sampling in deep, dimictic lakes are discussed.

Abstract Purpose Thousands of aquifers worldwide have been polluted by leachate from landfills and many more remained threatened. Microbial communities from these environments play a crucial role in mediating biodegradation and maintaining the biogeochemical cycles, but their co-occurrence and assembly mechanism have not been investigated. Method Here, we coupled network analysis with multivariate statistics to assess the relative importance of deterministic versus stochastic microbial assembly in an aquifer undergoing intrinsic remediation, using 16S metabarcoding data generated through Illumina MiSeq sequencing of the archaeal/bacterial V3–V4 hypervariable region. Results Results show that both the aquifer-wide and localised community co-occurrences deviate from expectations under null models, indicating the predominance of deterministic processes in shaping the microbial communities. Further, the amount of variation in the microbial community explained by the measured environmental variables was 55.3%, which illustrates the importance of causal factors in forming the structure of microbial communities in the aquifer. Based on the network topology, several putative keystone taxa were identified which varied remarkably among the wells in terms of their number and composition. They included Nitrospira, Nitrosomonadaceae, Patulibacter, Legionella, uncharacterised Chloroflexi, Vicinamibacteriales, Neisseriaceae, Gemmatimonadaceae, and Steroidobacteraceae. The putative keystone taxa may be providing crucial functions in the aquifer ranging from nitrogen cycling by Nitrospira, Nitrosomonadaceae, and Steroidobacteraceae, to phosphorous bioaccumulation by Gemmatimonadaceae. Conclusion Collectively, the findings provide answers to fundamental ecological questions which improve our understanding of the microbial ecology of landfill leachate plumes, an ecosystem that remains understudied.

Abstract Remediation of groundwater from landfill contamination presents a serious challenge due to the complex mixture of contaminants discharged from landfills. Here, we show the significance of a nature-based solution to a landfill-contaminated aquifer in southeast Norway. Groundwater physicochemical parameters monitored for twenty-eight years were used as a proxy to infer natural remediation. Results show that concentrations of the major chemical variables decreased with time and distance until they tailed off. An exception to this was sulphate, which showed an increase, but apparently, exhibits a stationary phase. The water types were found to be most similar between samples from active landfill and post-closure stages, while samples from the stabilised stage showed a different water type. All the chemical parameters of samples from the stabilised stage were found to be within the Norwegian drinking water standards, except iron and manganese, which were only marginally above the limits, an indication of a possible recovery of this aquifer. The findings highlight the significance of natural attenuation processes in remediating contaminated aquifers and have significant consequences for future contamination management, where natural remediation can be viewed as an alternative worth exploring. This is promising in the wake of calls for sustainable remediation management strategies.

GABAA receptor-positive modulators are well-known to induce sedation, sleep, and general anesthesia. Conversely, GABAA receptor negative allosteric modulators (GABAARNAMs) can increase arousal and induce seizures. Motivated by our studies with patients with hypersomnia, and our discovery that two GABAARNAMs can restore the Excitation/Inhibition (E/I) balance in vitro and arousal in vivo, we chose to screen 11 compounds that have been reported to modulate arousal, to see if they shared a GABA-related mechanism. We determined modulation with both conventional and microfluidic patch clamp methods. We found that receptor activation was variably modulated by all 11 compounds: Rifampicin (RIF), Metronidazole (MET), Minocycline (MIN), Erythromycin (ERY), Ofloxacin (OFX), Chloroquine (CQ), Hydroxychloroquine sulfate (HCQ), Flumazenil (FLZ), Pentylenetetrazol (PTZ), (-)-Epigallocatechin Gallate (EGCG), and clarithromycin (CLR). The computational modeling of modulator–receptor interactions predicted drug action at canonical binding sites and novel orphan sites on the receptor. Our findings suggest that multiple avenues of investigation are now open to investigate large and brain-penetrant molecules for the treatment of patients with diminished CNS E/I balance.

Abstract Background Candidatus Neoehrlichia mikurensis is an emerging tick-borne pathogen. It is widely distributed in Ixodes ricinus ticks in Europe, but knowledge of its distribution in Norway, where I. ricinus reaches its northern limit, is limited. In this study we have developed a real time PCR test for Ca. N. mikurensis and used it to investigate the distribution of Ca. N. mikurensis in Norway. Results Real time PCR targeting the groEL gene was developed and shown to be highly sensitive. It was used to detect Ca. N. mikurensis in 1651 I. ricinus nymphs and adults collected from twelve locations in Norway, from the eastern Oslo Fjord in the south to near the Arctic Circle in the north. The overall prevalence was 6.5% and varied locally between 0 and 16%. Prevalence in adults and nymphs was similar, suggesting that ticks acquire Ca. N. mikurensis predominantly during their first blood meal. In addition, 123 larvae were investigated; Ca. N. mikurensis was not found in larvae, suggesting that transovarial transmission is rare or absent. Sequence analysis suggests that a single variant dominates in Norway. Conclusions Ca. N. mikurensis is widespread and common in ticks in Norway and reaches up to their northern limit near the Arctic Circle. Ticks appear to acquire Ca. N. mikurensis during their first blood meal. No evidence for transovarial transmission was found.

Objective Abstract The genetic markers designed for this study can facilitate future genetic studies on the rock ptarmigan (Lagopus muta). To our knowledge no microsatellite markers have ever been developed specifically for this species before. These new microsatellite markers will be useful for population genetics studies and for future conservation projects. Results Using Next Generation Sequencing 6252 potential microsatellite sequences were found. Sixteen nonpalindromic tetranucleotide microsatellites and their respective primers were selected. The markers were tested on both the rock ptarmigan and the willow grouse (L. lagopus). The number of alleles varied between 2 and 18 for the rock ptarmigan, and between 3 and 13 for the willow grouse. Expected heterozygosity was in the range 0.1244–0.8692 and 0.1358–0.8722 for the rock ptarmigan and the willow grouse, respectively.

Dopamine (DA) loss in Parkinson’s disease (PD) alters the function of striatal projection neurons (SPNs) and causes motor deficits, but DA replacement can induce further abnormalities. A key pathological change in animal models and patients is SPN hyperactivity; however, the role of glutamate in altered DA responses remains elusive. We tested the effect of locally applied AMPAR or NMDAR antagonists on glutamatergic signaling in SPNs of parkinsonian primates. Following a reduction in basal hyperactivity by antagonists at either receptor, DA inputs induced SPN firing changes that were stable during the entire motor response, in clear contrast with the typically unstable effects. The SPN activity reduction over an extended putamenal area controlled the release of involuntary movements in the “on” state and therefore improved motor responses to DA replacement. These results demonstrate the pathophysiological role of upregulated SPN activity and support strategies to reduce striatal glutamate signaling for PD therapy.

Birds are important hosts for the first life stages of the Ixodes ricinus tick and they can transport their parasites over long distances. The aim of this study was to investigate the prevalence of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Neoehrlichia mikurensis and Rickettsia helvetica in ticks collected from migratory birds in Norway. A total of 815 Ixodes ricinus ticks from 216 birds trapped at Lista Bird Observatory in southern Norway during spring and autumn migration in 2008 were analysed by real-time PCR. B. burgdorferi s. l. was the most prevalent pathogen, detected in 6.1% of the ticks. The prevalence of N. mikurensis, A. phagocytophilum and R. helvetica was 1.2%, 0.9% and 0.4% respectively. In addition, one sample (0.1%) was positive for B. miyamotoi. In total, 8.2% of the ticks were infected with at least one pathogen. Co-infection with B. burgdorferi s. l. and N. mikurensis or A. phagocytophilum was found in 6.0% of the infected ticks. Our results show that all the known major tick-borne bacterial pathogens in Norway are subject to transport by migratory birds, potentially allowing spread to new areas. Our study showed a surprisingly high number of samples with PCR inhibition (57%). These samples had been extracted using standard methodology (phenol-chloroform extraction). This illustrates the need for inhibition controls to determine true prevalence rates.

The boundary between the boreal and arctic biomes in northwest Europe has been a matter of debate for many years. Some authors consider that the boundary is marked by the northern limit of tree growth in the northernmost Norwegian mainland. In this study we have collected air and soil temperature data from 37 heath stands from northern Finnmark (71°N), the northernmost part of the Norwegian mainland, through Bear Island (74°N) in the Barents sea, to Adventsdalen (78)°N (in Spitsbergen) in Svalbard archipelago. In Finnmark, plots both south and north of the treeline were investigated. Vegetation and soil chemistry analyses were performed on the plots in Finnmark and Svalbard. Significant decreasing south-north trends in air and soil temperatures were observed from Finnmark to Spitsbergen. Soils in Finnmark were acidic and rich in organic matter, while those on Adventsdalen were basic and poor in organic matter. Vegetational analysis identified five communities: three in Finnmark and two on Adventsdalen. The communities in Finnmark had marked mutual similarities but were very different from those on Adventsdalen. No significant ecological differences between heaths south and north of the treeline in Finnmark were observed. Air and soil temperature variables in Finnmark were outside the recognized range for the arctic biome and inconsistent with the presence of permafrost both south and north of the treeline. A major difference between Finnmark and Spitsbergen was amount of soil frost and length of the growing season. Our results suggest that the boreal biome extends all the way to the north coast of mainland Norway; and previously used division of heaths in Finnmark into boreal, alpine and arctic biomes is not justified.

As the world's human population increases, transformation of natural landscapes into urban habitats continues to increase. In Africa, rates of human population growth and urbanisation are among the highest in the world, but the impacts of these processes on the continent's biodiversity remain largely unexplored. Furthermore, the effects of ongoing anthropogenic climate change are likely to be severe and to interact with urbanisation. Some organisms appear resilient to urbanisation, and even proliferate in human-modified environments. One such species is the peregrine falcon Falco peregrinus in Cape Town, South Africa. Using a long-term data set (1989-2014), we investigate the relationship between breeding attempts, timing of breeding and breeding performance under varying weather conditions. Exploring these issues along an urbanisation gradient, we focus on the role of artificially provided nest boxes, and their capacity to buffer against extreme weather events. Pairs in more urbanised areas, and particularly those in nest boxes, were more likely to breed and to commence breeding earlier. Additionally, pairs using nest boxes were more likely to breed in years with higher rainfall. Warm and dry weather conditions generally advanced the timing of breeding, although this relationship with weather was not seen for urban pairs using nest boxes. Furthermore, weather did not impact breeding performance directly (breeding success and fledged brood size), but timing of breeding did, with earlier breeders producing more fledglings. Our study shows that falcons breeding in specially provided nest boxes were less sensitive to local weather dynamics than pairs using more natural nest sites. This has important implications as it suggests that the managed provision of such nesting sites can help this key urban species to cope with extreme weather events, which are predicted to increase with climate change.

After publication of our article [1] it came to our notice that the source of the sequence for the control plasmid, pNeo (Materials and methods: Controls) was incorrectly stated as AB094461. The correct accession number is AB074461. The authors apologize for any confusion this may have caused.

This paper draws on longitudinal birth cohort data for Britain to analyse participation in learning activities by people in their 30s and 40s. People in this age group have received less attention than either young adults or people in retirement. Yet technical change and the need for new skills make it important for them to engage in learning to improve their prospects at work. We investigate participation and non-participation in a range of different types of learning including gaining qualifications, vocational training and learning for interest. Statistical models explore how factors which occur before their 30s influence learning in this phase of the lifecourse. The results show that cumulative advantage is important -- those with the highest qualifications in early adulthood were most likely to engage in further learning later on. Participation in learning activities of any kind in young adulthood was also a key antecedent factor predicting higher chances of participating in learning in mid-life.

Interactions between different phytoplankton taxa and heterotrophic bacterial communities within aquatic environments can differentially support growth of various heterotrophic bacterial species. In this study, phytoplankton diversity was studied using traditional microscopic techniques and the bacterial communities associated with phytoplankton bloom were studied using High Throughput Sequencing (HTS) analysis of 16S rRNA gene amplicons from the V1-V3 and V3-V4 hypervariable regions. Samples were collected from Lake Akersvannet, a eutrophic lake in South Norway, during the growth season from June to August 2013. Microscopic examination revealed that the phytoplankton community was mostly represented by Cyanobacteria and the dinoflagellate Ceratium hirundinella. The HTS results revealed that Proteobacteria (Alpha, Beta, and Gamma), Bacteriodetes, Cyanobacteria, Actinobacteria and Verrucomicrobia dominated the bacterial community, with varying relative abundances throughout the sampling season. Species level identification of Cyanobacteria showed a mixed population of Aphanizomenon flos-aquae, Microcystis aeruginosa and Woronichinia naegeliana. A significant proportion of the microbial community was composed of unclassified taxa which might represent locally adapted freshwater bacterial groups. Comparison of cyanobacterial species composition from HTS and microscopy revealed quantitative discrepancies, indicating a need for cross validation of results. To our knowledge, this is the first study that uses HTS methods for studying the bacterial community associated with phytoplankton blooms in a Norwegian lake. The study demonstrates the value of considering results from multiple methods when studying bacterial communities.

Place can be generally defined as a location that has been assigned meaning through human experience, and as such it is of multidisciplinary scientific interest. Up to this point place has been studied primarily within the context of social sciences as a theoretical construct. The availability of large amounts of user-generated content, e.g. in the form of social media feeds or Wikipedia contributions, allows us for the first time to computationally analyze and quantify the shared meaning of place. By aggregating references to human activities within urban spaces we can observe the emergence of unique themes that characterize different locations, thus identifying places through their discernible sociocultural signatures. In this paper we present results from a novel quantitative approach to derive such sociocultural signatures from Twitter contributions and also from corresponding Wikipedia entries. By contrasting the two we show how particular thematic characteristics of places (referred to herein as platial themes) are emerging from such crowd-contributed content, allowing us to observe the meaning that the general public, either individually or collectively, is assigning to specific locations. Our approach leverages probabilistic topic modelling, semantic association, and spatial clustering to find locations are conveying a collective sense of place. Deriving and quantifying such meaning allows us to observe how people transform a location to a place and shape its characteristics.

Abstract DNA methylation variations are prevalent in human obesity but evidence of a causative role in disease pathogenesis is limited. Here, we combine epigenome-wide association and integrative genomics to investigate the impact of adipocyte DNA methylation variations in human obesity. We discover extensive DNA methylation changes that are robustly associated with obesity (N = 190 samples, 691 loci in subcutaneous and 173 loci in visceral adipocytes, P 500 target genes, and identify putative methylation-transcription factor interactions. Through Mendelian Randomisation, we infer causal effects of methylation on obesity and obesity-induced metabolic disturbances at 59 independent loci. Targeted methylation sequencing, CRISPR-activation and gene silencing in adipocytes, further identifies regional methylation variations, underlying regulatory elements and novel cellular metabolic effects. Our results indicate DNA methylation is an important determinant of human obesity and its metabolic complications, and reveal mechanisms through which altered methylation may impact adipocyte functions.

CT scans of an unnamed mummified adult from Egypt, now in the Chau Chak Wing Museum, University of Sydney (NMR.27.3), reveal it to be fully sheathed in a mud shell or carapace, exposing a mortuary treatment not previously documented in the Egyptian archaeological record. The carapace was placed between layers of linen wrappings thus it was not externally visible. Radiocarbon dating of textile samples provide a range of c.1370-1113 cal BC (95.4% probability), with a median date of 1207 cal BC. When assessed against mummification techniques of the era, the individual is placed in the late 19th-20th Dynasty, at the later end of this date range. Multi-proxy analysis including μ-XRF and Raman spectroscopy of carapace fragments from the head area revealed it to consist of three layers, comprising a thin base layer of mud, coated with a white calcite-based pigment and a red-painted surface of mixed composition. Whether the whole surface of the carapace was painted red is unknown. The carapace was a form of ancient conservation applied subsequent to post-mortem damage to the body, intended to reconfigure the body and enable continued existence of the deceased in the afterlife. The carapace can also be interpreted as a form of elite emulation imitating resin shells found within the wrappings of royal bodies from this period.