A Set of Diagnostic Tests for Detection of Active Babesia duncani Infection.

OBJECTIVES: Human babesiosis, an emerging potential fatal tick-borne disease caused by intraerythrocytic parasites of the Babesia genus. B. duncani is one of the Babesia species cause severe and life-threatening infections in humans. Detecting B. duncani infection is essential for accurate diagnosis and effective disease management. While molecular assays for detection in blood exist, there is still no reliable method to detect biomarkers of active infection. METHODS: We developed the first B. duncani antigen capture assays, targeting two immunodominant antigens, BdV234 and BdV38. These assays were validated using established in vitro and in vivo B. duncani-infection levels, both before and after therapy. RESULTS: The assay displayed no cross-reactivity with other species such as B. microti, B. divergens, Babesia MO1, or P. falciparum. It can detect as few as 115 infected erythrocytes/µl blood. Screening of 1,731 blood samples from various biorepositories, including samples previously identified as Lyme and/or B. microti positive, as well as new specimens from field mice, revealed no evidence of B. duncani infection and cross reactivity. CONCLUSION: These assays have potential applications, such as point-of-care testing for early detection of B. duncani in patients, field tests for screening reservoir hosts, and high-throughput screening of blood samples collected for transfusion.

Decoding protein-RNA interactions using CLIP-based methodologies.

Protein-RNA interactions are central to all RNA processing events, with pivotal roles in the regulation of gene expression and cellular functions. Dysregulation of these interactions has been increasingly linked to the pathogenesis of human diseases. High-throughput approaches to identify RNA-binding proteins and their binding sites on RNA - in particular, ultraviolet crosslinking followed by immunoprecipitation (CLIP) - have helped to map the RNA interactome, yielding transcriptome-wide protein-RNA atlases that have contributed to key mechanistic insights into gene expression and gene-regulatory networks. Here, we review these recent advances, explore the effects of cellular context on RNA binding, and discuss how these insights are shaping our understanding of cellular biology. We also review the potential therapeutic applications arising from new knowledge of protein-RNA interactions.

A unified metric of human immune health.

Immunological health has been challenging to characterize but could be defined as the absence of immune pathology. While shared features of some immune diseases and the concept of immunologic resilience based on age-independent adaptation to antigenic stimulation have been developed, general metrics of immune health and its utility for assessing clinically healthy individuals remain ill defined. Here we integrated transcriptomics, serum protein, peripheral immune cell frequency and clinical data from 228 patients with 22 monogenic conditions impacting key immunological pathways together with 42 age- and sex-matched healthy controls. Despite the high penetrance of monogenic lesions, differences between individuals in diverse immune parameters tended to dominate over those attributable to disease conditions or medication use. Unsupervised or supervised machine learning independently identified a score that distinguished healthy participants from patients with monogenic diseases, thus suggesting a quantitative immune health metric (IHM). In ten independent datasets, the IHM discriminated healthy from polygenic autoimmune and inflammatory disease states, marked aging in clinically healthy individuals, tracked disease activities and treatment responses in both immunological and nonimmunological diseases, and predicted age-dependent antibody responses to immunizations with different vaccines. This discriminatory power goes beyond that of the classical inflammatory biomarkers C-reactive protein and interleukin-6. Thus, deviations from health in diverse conditions, including aging, have shared systemic immune consequences, and we provide a web platform for calculating the IHM for other datasets, which could empower precision medicine.

Academic learning time in physical education (ALT-PE) during an adapted tennis program for children with autism spectrum disorders.

As the prevalence of autism spectrum disorder (ASD) increases, there is a growing need to develop physical activity interventions that address the behavioral challenges experienced by individuals with ASD. Physical education teachers have employed behavioral supports that add more structure and adapt the environment for individuals with ASD, which are associated with increased engagement for individuals with ASD during PE. The purpose of this study was to quantify motor engaged behaviors (i.e. motor appropriate (MA)), motor inappropriate (MI), motor supported (MS) during skill practice in 18 individuals with ASD (ages 7-19 years) participating in an adapted tennis program (ACEing Autism) using the Academic Learning Time in Physical Education (ALT-PE) instrument. Overall, the supports provided during the program may have enabled participants with ASD to spend more time in MA and MS than MI. Indeed, the participants spent over 50% of their time in MA during the program.

The pros and confs of IL-18 activation.

Interleukin-1 (IL-1) family cytokines are key immunological regulators that achieve their signaling prowess after post-translational proteolytic processing. In this issue of Immunity, Dong et al. reveal the structural consequences of this process on proinflammatory IL-18, demonstrating that pro-IL-18 and mature IL-18 are structurally distinct.

Pseudomonas aeruginosa impairs mitochondrial function and metabolism during infection of corneal epithelial cells.

Pseudomonas aeruginosa (PA) is a gram-negative opportunistic pathogen that can infect the cornea as a result of trauma or contact lens wear. In addition to their known energy producing role, mitochondria are important mediators of immune signaling and host defense. While certain pathogens have developed strategies to evade host defenses by modulating host mitochondrial dynamics and metabolism, the ability of PA to harness host cell mitochondria during corneal infection is unknown. Using a combination of biochemical and imaging techniques, we show that PA infection of corneal epithelial cells induced mitochondrial fission in a DRP1-dependent manner that preceded PINK1/Parkin and FUNDC1-mediated mitophagy. PA also impaired NADH-linked respiration through a reduction in complex 1. This corresponded to a decrease in metabolic pathways related to glycolysis and the TCA cycle. Metabolomics analysis further demonstrated an upregulation of the pentose phosphate pathway, arginine, purine, and pyrimidine metabolism in PA infected cells. These pathways may provide a key source of nucleotides, amino acids, and nitrogen for both the host cell and PA, in addition to antioxidant functions. Following treatment with gentamicin to kill all extracellular bacteria, metabolic flux analysis showed that corneal epithelial cells were able to restore mitochondrial function despite the continued presence of intracellular PA. Taken together, these data demonstrate that mitochondrial dysfunction and metabolic rewiring in host cells is triggered by extracellular PA, but once inside, PA requires healthy mitochondria to ensure host cell survival.

Perceptions about dementia clinical trials among underrepresented populations: A nationally representative survey of U.S. dementia caregivers.

Background: The research community has historically failed to enroll diverse groups of participants in dementia clinical trials. A unique aspect of dementia care research is the requirement of a study partner, who can attest to the care recipient's clinical and functional capacity. The aim of this study is to assess racial and ethnic differences and the importance of various trial considerations among dementia caregivers, in their decision to participate in clinical research as study partners. Method: We embedded a vignette about a hypothetical dementia clinical trial in a nationally representative survey of U.S. dementia caregivers, oversampling non-Hispanic Black and Hispanic caregivers. Dementia caregivers were asked about their willingness to participate in the trial with their care recipient and rated the importance of nine considerations in hypothetical decisions to participate. Caregiver demographic characteristics were analyzed as predictors of trial participation in a base demographic model. In a second reasons model caregiver demographic characteristics and the rated importance of the nine considerations were separately analyzed as predictors; both models used survey-weighted logistic regression. Result: The sample consisted of 610 dementia caregivers, including 156 non-Hispanic Black and 122 Hispanic caregiver participants. In the base demographic model, hypothetical trial participation was negatively associated with older caregiver age (OR (odds ratio) = 0.72, p = < 0.001). In the reasons model, the rated importance of a social responsibility to help others by participating in research was significantly associated with participation (OR = 1.56, p = 0.049), while the importance of the possibility of the care recipient experiencing serious side effects was negatively associated with participation (OR = 0.51, p = 0.003). In both models there was no significant difference in hypothetical participation between non-Hispanic Black and non-Hispanic White caregivers, or between Hispanic and non-Hispanic White caregivers. Conclusion: Hispanic and non-Hispanic Black dementia caregivers were not less likely than non-Hispanic White dementia caregivers to participate in a hypothetical dementia clinical trial. Our study suggests that failures to recruit diverse populations in dementia clinical research are not attributable to less willingness among members of underrepresented groups but may instead reflect structural barriers and historic exclusion from trial participation.

Evaluating foliar insecticides and economic thresholds for Tychius picirostris (Coleoptera: Curculionidae) management in Oregon white clover seed production.

The clover seed weevil, Tychius picirostris Fabricius (Coleoptera: Curculionidae), is a major pest in Oregon white clover seed crops. Reliance on synthetic pyrethroid insecticides and limited availability of diverse modes of action (MoAs) has increased insecticide resistance selection in regional T. picirostris populations, emphasizing the need to evaluate novel chemistries and rotational strategies for effective insecticide resistance management (IRM). The efficacy of 8 foliar insecticide formulations for managing T. picirostris adult and larval life stages was determined in small and large-plot field trials across 2 crop years. In both years, bifenthrin (Brigade 2EC), the grower's standard, showed negligible adult and larval suppression. Insecticide formulations with isocycloseram and cyantraniliprole active ingredients reduced adult and larval populations when applied at BBCH 59-60 (prebloom) and BBCH 65-66 (full bloom) growth stages, respectively. While differences in T. picirostris abundance were observed among insecticide treatments, seed yield differences were not detected in large-plot trials. Larval abundance was correlated with reduced seed yield, and an economic threshold of ≥3 larvae per 30 inflorescences was determined as a conservative larval threshold to justify foliar applications of diamide insecticides. Additional commercial white clover seed fields were surveyed to compare larval scouting techniques, including a standard Berlese funnel and a grower's do-it-yourself funnel. Both larval extraction techniques were correlated and provided similar estimates of larval abundance. These findings demonstrate new MoAs, optimal insecticide application timing, and larval monitoring methods that can be incorporated into an effective T. picirostris IRM program in white clover seed crops.

The Cannabis Gray Market: A Case for Cannabis Regulatory Science Research.

The cannabis gray market poses significant public health concerns and remains a major threat to consumer and/or potential consumer uptake of regulated cannabis markets in jurisdictions with legal state-sponsored cannabis programs. In this perspective, we provide an overview of the cannabis gray market, and describe an integrated epidemiological and regulatory science framework to study the gray market. Using tobacco regulatory science as a guide, we introduce example cannabis regulatory science research activities as a means to improve the field's understanding of the cannabis gray market. Cannabis regulatory science is a developing field that can improve our understanding of the cannabis regulatory ecosystem and provide regulatory officials and policymakers alike with much needed data to inform regulatory decision-making and improve the success and uptake of state-sponsored cannabis programs.

Pseudomonas aeruginosa infection increases palmitoyl carnitine release by host-derived extracellular vesicles.

Pseudomonas aeruginosa (PA), an opportunistic gram-negative pathogen, is the most common pathogen identified in all culture positive cases of infectious keratitis. Extracellular vesicles (EVs) are released by most cells in the body and function in intercellular communication. We have previously reported a change in the proteome of host-derived EVs from corneal epithelial cells during PA infection. In the present study, we investigated changes in the metabolome of host-derived EVs from PA infected (PA-C EVs) and non-infected cells (C EVs). We found that one metabolite, palmitoyl carnitine (PAMC), was significantly upregulated in PA-C EVs. To determine the significance of PAMC release, we investigated the effect of PAMC treatment on corneal epithelial cells and neutrophils. EVs were isolated from culture media using size exclusion chromatography. EVs were then characterized using nanoparticle tracking analysis, transmission electron microscopy, and western blot. Metabolomics was performed using an untargeted approach. We found that palmitoyl carnitine (PAMC) was the most abundant metabolite present in PA-C EVs and was increased more than 3 fold compared to C EVs. Treatment of corneal epithelial cells with increasing levels of PAMC increased nuclear translocation of the NF-κB subunit p65. This was associated with an increase in IL-8 production and neutrophil migration. PAMC also increased levels of mitochondrial calcium. Upon inoculation of corneal epithelial cells with PA, 50 µM PAMC completely eradicated intracellular PA, but stimulated growth of extracellular PA. Taken together, these findings suggest that PA exploits EV release by host cells to deplete PAMC from the intracellular environment.

CryoET reveals actin filaments within platelet microtubules.

Crosstalk between the actin and microtubule cytoskeletons is important for many cellular processes. Recent studies have shown that microtubules and F-actin can assemble to form a composite structure where F-actin occupies the microtubule lumen. Whether these cytoskeletal hybrids exist in physiological settings and how they are formed is unclear. Here, we show that the short-crossover Class I actin filament previously identified inside microtubules in human HAP1 cells is cofilin-bound F-actin. Lumenal F-actin can be reconstituted in vitro, but cofilin is not essential. Moreover, actin filaments with both cofilin-bound and canonical morphologies reside within human platelet microtubules under physiological conditions. We propose that stress placed upon the microtubule network during motor-driven microtubule looping and sliding may facilitate the incorporation of actin into microtubules.

Long-Term Kidney Outcomes after Pediatric Acute Kidney Injury.

BACKGROUND: Acute kidney injury (AKI) is common in hospitalized children. Pediatric AKI receiving acute kidney replacement therapy (KRT) is associated with long-term chronic kidney disease (CKD), hypertension, and death. We aim to determine the outcomes after AKI in children who did not receive acute KRT, since these remain uncertain. METHODS: Retrospective cohort study of all hospitalized children (0-18 years) surviving AKI without acute KRT between 1996-2020 in Ontario, Canada, identified by validated diagnostic codes in provincial administrative health databases. Children with prior KRT, CKD, or AKI were excluded. Cases were matched with up to four hospitalized comparators without AKI by age, neonatal status, sex, intensive care unit admission, cardiac surgery, malignancy, hypertension, hospitalization era, and a propensity score for AKI. Patients were followed until death, provincial emigration, or censoring in March 2021. The primary outcome was long-term major adverse kidney events (MAKE-LT; a composite of all-cause mortality, long-term KRT, or incident CKD). RESULTS: We matched 4,173 pediatric AKI survivors with 16,337 hospitalized comparators. Baseline covariates were well-balanced following propensity score matching. During median 9.7-year follow-up, 18% of AKI survivors developed MAKE-LT vs. 5% of hospitalized comparators (hazard ratio [HR] 4.0, 95% confidence interval [CI] 3.6-4.4). AKI survivors had higher rates of long-term KRT (2% vs. <1%; HR 11.7, 95%CI 7.5-18.4), incident CKD (16% vs. 2%; HR 7.9, 95%CI 6.9-9.1), incident hypertension (17% vs. 8%; HR 2.3, 95%CI 2.1-2.6), and AKI during subsequent hospitalization (6% vs. 2%; HR 3.7, 95%CI 3.1-4.5), but no difference in all-cause mortality (3% vs. 3%; HR 0.9, 95%CI 0.7-1.1). CONCLUSIONS: Children surviving AKI without acute KRT were at higher long-term risk of CKD, long-term KRT, hypertension, and subsequent AKI vs. hospitalized comparators.

Different trajectories of relationship satisfaction among rural Black couples following preventive relationship intervention.

OBJECTIVE: This study examined variability in response to a couple and relationship education program for Black coparenting couples using group-based trajectory modeling. We identified groups of couples with different relationship satisfaction trajectories across a 2-year period following preintervention assessment. METHOD: Black couples with a preadolescent child were randomized to the Protecting Strong African American Families intervention (N = 170) or a control condition (N = 174) and reported on their relationship satisfaction preintervention and approximately 9, 17, and 25 months later. RESULTS: Results indicated two trajectory groups among men and women receiving the intervention, comprising a large group consistently high in relationship satisfaction over time and a smaller group with moderate, stable satisfaction over time. In the control condition, there was also a large group of men and women who remained consistently high in satisfaction, but the smaller group with initially moderate satisfaction experienced decreased satisfaction over time. Trajectories among men and women in the moderate groups differed by treatment condition, reflecting nonsignificant change (i.e., stable satisfaction) for those receiving intervention and significant decline for controls. Trajectories among women in the high group also differed by treatment condition, reflecting better functioning among women receiving intervention. CONCLUSIONS: Results indicated many benefits of intervention, most notably the stabilization of satisfaction among couples beginning with moderate satisfaction (who experienced declining satisfaction in the control condition). Similar examination of within-sample variability in response to other couple and relationship education programs may enhance understanding of specific treatment effects and guide identification of those most likely to benefit from relationship intervention. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

The mitochondrial calcium uniporter: Balancing tumourigenic and anti-tumourigenic responses.

Increased malignancy and poor treatability associated with solid tumour cancers have commonly been attributed to mitochondrial calcium (Ca2+) dysregulation. The mitochondrial Ca2+ uniporter complex (mtCU) is the predominant mode of Ca2+ uptake into the mitochondrial matrix. The main components of mtCU are the pore-forming mitochondrial Ca2+ uniporter (MCU) subunit, MCU dominant-negative beta (MCUb) subunit, essential MCU regulator (EMRE) and the gatekeeping mitochondrial Ca2+ uptake 1 and 2 (MICU1 and MICU2) proteins. In this review, we describe mtCU-mediated mitochondrial Ca2+ dysregulation in solid tumour cancer types, finding enhanced mtCU activity observed in colorectal cancer, breast cancer, oral squamous cell carcinoma, pancreatic cancer, hepatocellular carcinoma and embryonal rhabdomyosarcoma. By contrast, decreased mtCU activity is associated with melanoma, whereas the nature of mtCU dysregulation remains unclear in glioblastoma. Furthermore, we show that numerous polymorphisms associated with cancer may alter phosphorylation sites on the pore forming MCU and MCUb subunits, which cluster at interfaces with EMRE. We highlight downstream/upstream biomolecular modulators of MCU and MCUb that alter mtCU-mediated mitochondrial Ca2+ uptake and may be used as biomarkers or to aid in the development of novel cancer therapeutics. Additionally, we provide an overview of the current small molecule inhibitors of mtCU that interact with the Asp residue of the critical Asp-Ile-Met-Glu motif or through other allosteric regulatory mechanisms to block Ca2+ permeation. Finally, we describe the relationship between MCU- and MCUb-mediating microRNAs and mitochondrial Ca2+ uptake that should be considered in the discovery of new treatment approaches for cancer.

Organic phosphate but not inorganic phosphate regulates Fgf23 expression through MAPK and TGF-ꞵ signaling.

One of the main regulators of phosphate homeostasis is fibroblast growth factor 23 (FGF23), secreted by osteocytes. The effects of organic versus inorganic dietary phosphate on this homeostasis are unclear. This study used MC3T3-E1 FGF23-producing cells to examine the transcriptomic responses to these phosphates. Most importantly, the expression and secretion of FGF23 were only increased in response to organic phosphate. Gene ontology terms related to a response to environmental change were only enriched in cells treated with organic phosphate while cells treated with inorganic phosphate were enriched for terms associated with regulation of cellular phosphate metabolism. Inhibition of MAPK signaling diminished the response of Fgf23 to organic phosphate, suggesting it activates FGF23. TGF-ß signaling inhibition increased Fgf23 expression after the addition of organic phosphate, while the negative TGF-ß regulator Skil decreased this response. In summary, the observed differential response of FGF23-producing to phosphate types may have consequences for phosphate homeostasis.

There's an App for That: Exploring the Market for Contraceptive Fertility Tracking Apps in the Philippines.

For generations, women have relied on fertility awareness methods to plan and prevent pregnancy, for over a decade, many have been aided by digital tools to do so. New contraceptive fertility tracking apps (CFTAs)-that are backed by clinical efficacy trials to support their effectiveness as contraception-have the potential to enhance method choice and offer users a unique contraceptive option, but there is little evidence to inform the decisions around expanding access, particularly in low-and middle-income countries. We conducted a mixed methods study with quantitative online surveys (n = 1600) and qualitative interviews (n = 36) to explore the potential appeal of and demand for a hypothetical CFTA in one such market, the Philippines. Interest in using a CFTA was high among our Internet-engaged, urban study population, with 83.9% "definitely" or "probably" interested in using it. Across demographic profiles, respondents perceived the appeal of the method as "natural" and "convenient." A majority were willing to pay for the method, though notably at a price (5.20 USD) below that of currently available CFTAs. We discuss various important factors to be considered before bringing a method like this to new markets, including the potential implications of equity constraints in reaching a wider market and the unexpected prevalence of other period-tracking apps not intended as contraception being used in this market that could complicate any future roll-out. These issues could be explored further with additional research.

Prevalence of current chronic pain in Royal Canadian Mounted Police cadets.

Background: Nearly half of active duty Royal Canadian Mounted Police (RCMP) officers report experiencing current chronic pain (43%; i.e. pain lasting longer than 3 months). Most RCMP officers who report chronic pain indicate that the pain started after working as RCMP officers (91%). Baseline data on chronic pain prevalence among RCMP cadets has not been available. Aims: The current study was designed to provide cross-sectional estimates of chronic pain prevalence among RCMP cadets starting the Cadet Training Program and to assess for sociodemographic differences among participants. Methods: The RCMP Study uses a longitudinal prospective sequential experimental cohort design to create a clustered randomized trial that engages individual participants for 5.5 years. The current article provides cross-sectional associations between chronic pain prevalence and sociodemographic characteristics. Participants were RCMP cadets starting the Cadet Training Program (n = 770). Location, intensity (on a 0-10 scale and days per week experienced), and duration (number of months) of chronic pain were reported. Differences across sociodemographic characteristics were examined. Results: Few RCMP cadets reported experiencing chronic pain (10%); lower back pain was rated as the most severe in terms of intensity and duration and second most frequently reported in number of days experienced per week. Prevalence of chronic pain was lower among RCMP cadets than among RCMP officers. Conclusions: Chronic pain prevalence among active duty RCMP officers may result from or be moderated by operational duties, as well as routine aging. Future researchers could examine ways to mitigate chronic pain development during RCMP officer careers.

Contexte: Près de la moitié des agents de la Gendarmerie royale du Canada (GRC) en service actif déclarent souffrir de douleur chronique (43 %; c'est-à-dire une douleur qui dure plus de trois mois). La plupart des agents de la GRC qui déclarent souffrir de douleur chronique indiquent que la douleur a commencé après avoir travaillé comme agents de la GRC (91 %). Il n'existe pas de données de référence sur la prévalence de la douleur chronique chez les cadets de la GRC.Objectifs: La présente étude a été conçue pour fournir des estimations transversales de la prévalence de la douleur chronique chez les cadets de la GRC qui commencent le Programme de formation des cadets et évaluer les différences sociodémographiques entre les participants.Méthodes: L'étude sur la GRC utilise un devis de cohorte expérimental séquentiel prospectif longitudinal pour créer un essai randomisé en grappes impliquant des participants individuels pendant 5,5 ans. Le présent article présente des associations transversales entre la prévalence de la douleur chronique et les caractéristiques sociodémographiques. Les participants étaient des cadets de la GRC qui commençaient le Programme de formation des cadets (n = 770). Le lieu, l'intensité (sur une échelle de 0 à 10 et selon le nombre de jours par semaine où la douleur était ressentie), de même que la durée (nombre de mois) pendant laquelle la douleur chronique était ressentie, ont été déclarés.Résultats: Peu de cadets de la GRC ont déclaré souffrir de douleur chronique (10 %); la douleur lombaire a été évaluée comme la plus sévère en termes d'intensité et de durée, et la deuxième la plus fréquemment rapportée en nombre de jours par semaine. La prévalence de la douleur chronique était plus faible chez les cadets de la GRC que chez les agents de la GRC.Conclusions: La prévalence de la douleur chronique chez les agents de la GRC en service actif peut résulter des tâches opérationnelles ou être modérée par celles-ci, ainsi que par le vieillissement normal. Les futurs chercheurs pourraient examiner les moyens d'atténuer le développement de la douleur chronique au cours de la carrière des agents de la GRC.

Machine learning with taxonomic family delimitation aids in the classification of ephemeral beaked whale events in passive acoustic monitoring.

Passive acoustic monitoring is an essential tool for studying beaked whale populations. This approach can monitor elusive and pelagic species, but the volume of data it generates has overwhelmed researchers' ability to quantify species occurrence for effective conservation and management efforts. Automation of data processing is crucial, and machine learning algorithms can rapidly identify species using their sounds. Beaked whale acoustic events, often infrequent and ephemeral, can be missed when co-occurring with signals of more abundant, and acoustically active species that dominate acoustic recordings. Prior efforts on large-scale classification of beaked whale signals with deep neural networks (DNNs) have approached the class as one of many classes, including other odontocete species and anthropogenic signals. That approach tends to miss ephemeral events in favor of more common and dominant classes. Here, we describe a DNN method for improved classification of beaked whale species using an extensive dataset from the western North Atlantic. We demonstrate that by training a DNN to focus on the taxonomic family of beaked whales, ephemeral events were correctly and efficiently identified to species, even with few echolocation clicks. By retrieving ephemeral events, this method can support improved estimation of beaked whale occurrence in regions of high odontocete acoustic activity.