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Abstract Self-replicating RNA (srRNA) technology, in comparison to mRNA vaccines, has shown dose-sparing by approximately 10-fold and more durable immune responses. However, no improvements are observed in the adverse events profile. Here, we develop an srRNA vaccine platform with optimized non-coding regions and demonstrate immunogenicity and safety in preclinical and clinical development. Optimized srRNA vaccines generate protective immunity (according to the WHO defined thresholds) at doses up to 1,000,000-fold lower than mRNA in female mouse models of influenza and rabies. Clinically, safety and immunogenicity of RBI-4000, an srRNA vector encoding the rabies glycoprotein, was evaluated in a Phase I study (NCT06048770). RBI-4000 was able to elicit de novo protective immunity in the majority of healthy participants when administered at a dose of 0.1, 1, or 10 microgram (71%, 94%, 100%, respectively) in a prime-boost schedule. Similarly, we observe immunity above the WHO benchmark of protection following a single administration in most participants at both 1 and 10 microgram doses. There are no serious adverse events reported across all cohorts. These data establish the high therapeutic index of optimized srRNA vectors, demonstrating feasibility of both low dose and single dose approaches for vaccine applications.

Surveillance of antimicrobial resistance in Kenya, Uganda, and Jordan identified multidrug-resistant high-risk bacterial clones: Escherichia coli sequence types 131, 1193, 69, 167, 10, 648, 410, 405 and Klebsiella pneumoniae sequence types 14, 147, 307, 258. Clones emerging in those countries exhibited high resistance mechanism diversity, highlighting a serious threat for multidrug resistance.

The Global Emerging Infections Surveillance (GEIS) program is the only Department of Defense (DoD) organization that coordinates global surveillance for emerging infectious diseases that affect US military forces operating in the United States or foreign locations. Since 1997, the GEIS program has focused on surveilling pathogens likely to affect military operations and the health of service members. The foundation of the GEIS program is the long-standing, mutually beneficial relationships between the DoD overseas laboratories and their host-country partners and militaries. Through centralized programmatic support, the GEIS program provides the infrastructure needed for a rapid and scalable response to emerging threats. The GEIS program continues to enhance and evolve its initiatives to provide timely, reliable information to decision-makers in the DoD. The GEIS program has been and will continue to be a vital source of actionable biosurveillance information during infectious disease events of global public health concern.

Travelers’ diarrhea has a high incidence rate among deployed US military personnel and can hinder operational readiness. The Global Travelers’ Diarrhea study is a US Department of Defense­–funded multisite surveillance effort to investigate the etiology and epidemiology of travelers’ diarrhea. During 2018–2023, we enrolled 512 participants at partner institutions in 6 countries: Djibouti, Georgia, Egypt, Honduras, Nepal, and Peru. Harmonized laboratory methods conducted at each partner institution identified >1 pathogens, including Escherichia coli (67%–82%), norovirus (4%–29%), and Campylobacter jejuni (2%–20%), in 403 (79%) cases. Among cases, 79.7% were single infections, 19.6% were double infections, and 0.7% were triple infections. The most common enterotoxigenic E. coli colonization factors identified were CS3 (25%) and CS21 (25%), followed by CS2 (18%) and CS6 (15%). These data can inform best treatment practices for travelers’ diarrhea and support US military health readiness.

Since its establishment in 1997, the US Department of Defense (DoD) Global Emerging Infections Surveillance (GEIS) program has provided support for infectious disease pandemic preparedness and response. The GEIS program has shown the value of having a central hub responsible for coordinating a global network of DoD laboratories that conduct surveillance for militarily relevant infectious disease threats. The program has supported the establishment and maintenance of capabilities for collecting, characterizing, and reporting on major infectious disease events, including the COVID-19 pandemic and mpox outbreak. The GEIS program enables the US government to mitigate infectious disease threats to DoD mission readiness and to effectively respond to pathogens worldwide. Continued investment in maintaining the GEIS program and its network is critical for timely detection and response to future emerging infectious disease threats in various populations within locations where gaps in US government or host-nation surveillance might exist.

Norovirus is the leading cause of acute gastroenteritis (AGE) worldwide. Norovirus outbreaks at military facilities can cause loss of training and working days and increased healthcare utilization, affecting force readiness. During 2013–2023, we enrolled 2,304 US military recruits from 4 basic training facilities to investigate AGE outbreaks among this population. Among enrollees, we detected norovirus in 433 (18.8%) AGE cases, and norovirus caused 49 AGE outbreaks during our longitudinal study. On average, each norovirus case-patient missed 1.2 training days due to illness, and 6.2% required infusion care. Whole-genome sequencing of selected samples from each outbreak produced full-length genomes (6,989–7,787 bp) for 39 samples. Norovirus GII.4 Sydney was the most (12/39, 30.8%) identified genotype over the study period. Phylogenetic and comparative genomic analyses revealed that several outbreak strains were responsible for causing >1 outbreak, even across different training sites. Our findings can inform infection control practices at military installations and overall norovirus vaccine development.

The rapid emergence of antimicrobial-resistant strains of Neisseria gonorrhoeae threatens treatment options and control efforts. The Uniformed Services University Gonococcal Reference Laboratory and Repository of the Global Emerging Infections Surveillance Program receives isolates from several geographically distinct regions worldwide. We analyzed 962 isolates collected during 2014–2022 for genomic and phenotypic antimicrobial resistance. Resistance to antimicrobial drugs previously used for gonococcal infections was high, but of most concern were increases of resistance to currently used antibiotic drugs, such as extended-spectrum cephalosporins and the alternative antibiotic treatment gentamicin. The percentage of isolates with reduced susceptibility to ceftriaxone was 3.6%, to cefixime was 2.5%, and to gentamicin was 15.0%. Although isolates were collected from populations of limited diversity, 706 (73.4%) of isolates demonstrated novel multiantigen sequence types, and 225 (23.4%) had novel multilocus sequence types. Continued surveillance of N. gonorrhoeae is essential to monitoring the prevalence and spread of resistant organisms worldwide.

Escherichia coli and Shigella spp. are leading bacterial causes of acute diarrhea in sub-Saharan Africa and pose risks to global communities, travelers, and the US military. Increasing antimicrobial resistance (AMR) in those and other enteric pathogens creates treatment challenges for clinicians. Inappropriate use of antimicrobial drugs, such as azithromycin for viral respiratory infections, increased during the COVID-19 pandemic. We evaluated AMR trends of 116 E. coli and 109 Shigella spp. isolates obtained from 1,672 pre–COVID-19 (2017–2019) and 1,118 post–COVID-19 (2022–2023) human fecal samples from Kenya. Azithromycin resistance increased significantly from before to after COVID-19, from 6.3% to 40.4% (p = 0.001). Phenotypic AMR profiles from a subset of isolates were compared with genotypic AMR information derived from whole genome sequencing. The most common AMR gene detected was the macrolide mph(A) gene. This study highlights the need for continued AMR surveillance.

Many higher educational institutions offer existing travel abroad study experiences. On the rise are short-term domestic travel experiences with the large cultural diversity present in the United States. As accessibility to study abroad programing is continuously faced with barriers such as cost, perceived safety, opportunity cost, and language, could domestic study programs offer equitable alternatives? With leadership experience being a necessary and sought after skill by students and employers, to what capacity does domestic travel provide participants leadership development? This autophotography study utilized photo-caption data to investigate the perceived leadership competencies developed by state FFA officers during a short-term domestic travel experience. Participant photo-caption responses communicated limited leadership competency development. Rather, participants conflated agricultural experiential exposure and gained social capital as perceived leadership competency development. Recommendations include developing leadership curriculum grounded in theory for both domestic and abroad travel experiences, front loading leadership competency development goals to participants, introducing post-trip interviews in future studies, and replicating the study within a global context in a travel experience abroad to understand perceptions beyond a domestic experience.

Although DNA methylation (DNAm) has been implicated in the pathogenesis of numerous complex diseases, from cancer to cardiovascular disease to autoimmune disease, the exact methylation sites that play key roles in these processes remain elusive. One strategy to identify putative causal CpG sites and enhance disease etiology understanding is to conduct methylome-wide association studies (MWASs), in which predicted DNA methylation that is associated with complex diseases can be identified. However, current MWAS models are primarily trained using the data from single studies, thereby limiting the methylation prediction accuracy and the power of subsequent association studies. Here, we introduce a new resource, MWAS Imputing Methylome Obliging Summary-level mQTLs and Associated LD matrices (MIMOSA), a set of models that substantially improve the prediction accuracy of DNA methylation and subsequent MWAS power through the use of a large summary-level mQTL dataset provided by the Genetics of DNA Methylation Consortium (GoDMC). Through the analyses of GWAS (genome-wide association study) summary statistics for 28 complex traits and diseases, we demonstrate that MIMOSA considerably increases the accuracy of DNA methylation prediction in whole blood, crafts fruitful prediction models for low heritability CpG sites, and determines markedly more CpG site-phenotype associations than preceding methods. Finally, we use MIMOSA to conduct a case study on high cholesterol, pinpointing 146 putatively causal CpG sites.

Schwannomas are benign neoplasms that arise from peripheral nerve sheaths. Typically found in peripheral nerves of the head, neck, and extremities, these tumors seldom arise in the retroperitoneum. We report the case of a 50-year-old male with a 5 cm right adrenal mass removed via robotic-assisted laparoscopic approach due to concerns for adrenocortical carcinoma, which surgical pathology revealed to be a rare adrenal schwannoma. This is the reported case.

Wildfires drastically impact the soil environment, altering the soil organic matter, forming pyrolyzed compounds, and markedly reducing the diversity of microorganisms. Pyrophilous fungi, especially the species from the orders Pezizales and Agaricales, are fire-responsive fungal colonizers of post-fire soil that have historically been found fruiting on burned soil and thus may encode mechanisms of processing these compounds in their genomes. Pyrophilous fungi are diverse. In this work, we explored this diversity and sequenced six new genomes of pyrophilous Pezizales fungi isolated after the 2013 Rim Fire near Yosemite Park in California, USA: Pyronema domesticum, Pyronema omphalodes, Tricharina praecox, Geopyxis carbonaria, Morchella snyderi, and Peziza echinospora. A comparative genomics analysis revealed the enrichment of gene families involved in responses to stress and the degradation of pyrolyzed organic matter. In addition, we found that both protein sequence lengths and G + C content in the third base of codons (GC3) in pyrophilous fungi fall between those in mesophilic/nonpyrophilous and thermophilic fungi. A comparative transcriptome analysis of P. domesticum under two conditions - growing on charcoal, and during sexual development - identified modules of genes that are co-expressed in the charcoal and light-induced sexual development conditions. In addition, environmental sensors such as transcription factors STE12, LreA, LreB, VosA, and EsdC were upregulated in the charcoal condition. Taken together, these results highlight genomic adaptations of pyrophilous fungi and indicate a potential connection between charcoal tolerance and fruiting body formation in P. domesticum.

Human pluripotent stem cells (hPSCs) are an exciting and promising source to enable cell replacement therapies for a variety of unmet medical needs. Though hPSCs can be successfully derived into numerous physiologically relevant cell types, effective translation to the clinic is limited by challenges in scalable production of high-quality cells, cellular immaturity following the differentiation process, and the use of animal-derived components in culture. To address these limitations, we have developed a fully defined, reproducible, and tunable thermoreversible polymer for high-quality, scalable 3D cell production. Our reproducible synthesis method enables precise control of gelation temperature (24°C-32°C), hydrogel stiffness (100-4000 Pa), and the prevention of any unintended covalent crosslinking. After material optimization, we demonstrated hPSC expansion, pluripotency maintenance, and differentiation into numerous lineages within the hydrogel. Overall, this 3D thermoreversible hydrogel platform has broad applications in scalable, high-quality cell production to overcome the biomanufacturing burden of stem cell therapy.

Autistic persons exhibit variable movement, loading, and coordination patterns during walking. While much research has examined walking, little to no research exists regarding running for autistic persons despite its prevalence and benefits as a mode of physical activity. This study determined if autistic adolescents demonstrate increased coordination variability during running compared to matched controls. Seventeen autistic adolescents (aged = 13-18 years) and seventeen sex, age, and body mass index matched controls performed running at two matched speeds: self-selected of autistic adolescents and at 3.0 m/s. Modified vector coding was used to determine the patterns of movement for foot-shank, shank-thigh, left/right thigh, and contralateral arm-thigh coupling. Coordination variability, measuring cycle-to-cycle variability, was determined during loading response and pushoff phases. Mixed-model analyses of variance were used to determine group by speed interactions and main effects. Coordination variability was nearly 2× larger (all p < 0.001) in autistic adolescents compared to controls. Speed main effects were found for several sagittal plane couples during loading response. In agreement with walking analyses, this study illustrates that autistic adolescents run with increased intra-limb, inter-limb, and cross-body coordination variability. Like walking, increased coordination variability during running may negatively impact this mode of physical activity for autistic persons.

Kennedy B Nkhoma,1 Lindsay Farrant,2 Olona Mzimkulu,2 Joy Hunter,2 Irene Higginson,1 Wei Gao,2 Matthew Maddocks,1 Liz Gwyther,2 Richard Harding1 1Cicely Saunders Institute of Palliative Care Policy and Rehabilitation, King’s College London, London, UK; 2Division of Interdisciplinary Palliative Care & Medicine, Department of Public Health and Family Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South AfricaCorrespondence: Kennedy B Nkhoma, Email kennedy.nkhoma@kcl.ac.ukBackground: Many deaths globally are attributable to non-communicable disease, and four-fifths of these deaths are in low- and middle-income countries. Globally, COPD is currently the third leading cause of mortality.Research Question: 1) To determine the prevalence and burden of symptoms and concerns, and 2) determine predicting factors of symptom burden among patients with COPD.Methods: A cross-sectional survey was conducted at eight primary care sites in Western Cape. We collected socio-demographic data (age, gender, smoking status, number of missed doses of prescribed medication in the last seven days) and clinical data (PEF and KPS). The Memorial Symptom Assessment Scale (MSAS), the Medical Outcomes Study, Social Support Survey (MOS-SSS), the London Chest Activity of Daily Living Scale (LCADLS) and the COPD Assessment Test (CAT) (impairment on person’s life) were administered to patients. We conducted ordered logistic regression analysis to assess factors associated with the burden of symptoms. MSAS subscales: 1) Global symptom distress index, 2) physical symptom distress and 3) psychological symptom distress were dependent outcomes. We constructed three ordinal logistic regression models for each of the three subscales. Covariates were MOS-SSS, LCADLS, CAT, demographic and clinical variables.Results: We recruited n=387 patients, mean age 59.5 years, 53.0% female. In multivariate analysis, each of the three models (ie, global, psychological and physical symptom distress) was positively associated with impairment on person’s life p< 0.001, difficulty to perform activities of daily living p< 0.001, and low social support p< 0.001. Old age was associated with lower global symptom distress (p=0.004), psychological and (0.014) physical distress (0.005). Missing 1 or more doses of medication was associated with higher levels of global (0.004) and physical (0.005) symptom distress.Interpretation: The high prevalence and burden of physical and psychological symptoms provides strong evidence of the need for integrating person-centred assessment and management of symptoms in primary care settings.Keywords: chronic lung disease, chronic obstructive pulmonary disease, symptom burden, symptom distress, symptom prevalence

To promote collaboration across canine science, address replicability issues, and advance open science practices within animal cognition, we have launched the ManyDogs consortium, modeled on similar ManyX projects in other fields. We aimed to create a collaborative network that (a) uses large, diverse samples to investigate and replicate findings, (b) promotes open science practices of pre-registering hypotheses, methods, and analysis plans, (c) investigates the influence of differences across populations and breeds, and (d) examines how different research methods and testing environments influence the robustness of results. Our first study combines a phenomenon that appears to be highly reliable—dogs’ ability to follow human pointing—with a question that remains controversial: do dogs interpret pointing as a social communicative gesture or as a simple associative cue? We collected data (N = 455) from 20 research sites on two conditions of a 2-alternative object choice task: (1) Ostensive (pointing to a baited cup after making eye-contact and saying the dog’s name); (2) Non-ostensive (pointing without eye-contact, after a throat-clearing auditory control cue). Comparing performance between conditions, while both were significantly above chance, there was no significant difference in dogs’ responses. This result was consistent across sites. Further, we found that dogs followed contralateral, momentary pointing at lower rates than has been reported in prior research, suggesting that there are limits to the robustness of point-following behavior: not all pointing styles are equally likely to elicit a response. Together, these findings underscore the important role of procedural details in study design and the broader need for replication studies in canine science.

The effects of immunodeficiency associated with chronic HIV infection on COVID-19 disease and viral persistence have not been directly addressed in a controlled setting. In this pilot study, we exposed two pigtail macaques (PTMs) chronically infected with SIVmac239, exhibiting from very low to no CD4 T cells across all compartments, to SARS-CoV-2. We monitored the disease progression, viral replication, and evolution, and compared these outcomes with SIV-naïve PTMs infected with SARS-CoV-2. No overt signs of COVID-19 disease were observed in either animal, and the SARS-CoV-2 viral kinetics and evolution in the SIVmac239 PTMs were indistinguishable from those in the SIV-naïve PTMs in all sampled mucosal sites. However, the single-cell RNA sequencing of bronchoalveolar lavage cells revealed an infiltration of functionally inert monocytes after SARS-CoV-2 infection. Critically, neither of the SIV-infected PTMs mounted detectable anti-SARS-CoV-2 T-cell responses nor anti-SARS-CoV-2 binding or neutralizing antibodies. Thus, HIV-induced immunodeficiency alone may not be sufficient to drive the emergence of novel viral variants but may remove the ability of infected individuals to mount adaptive immune responses against SARS-CoV-2.

IntroductionMortality in pediatric cerebral malaria (CM) in low- and middle-income countries (LMICs) is associated with brain swelling on magnetic resonance imaging (MRI); however, MRI is unavailable in most LMICs. Optic nerve sheath diameter (ONSD) measurement is an inexpensive method of detecting increased intracranial pressure compared with the invasive opening pressure (OP). Our primary objective was to determine if increased ONSD correlated with brain swelling on MRI in pediatric CM. Our secondary objective was to determine if increased ONSD correlated with increased OP and/or poor neurological outcome in pediatric CM. We hypothesized that increased ONSD would correlate with brain swelling on MRI and increased OP and that ONSD would be higher in survivors with sequelae and non-survivors.MethodsWe performed a retrospective chart review of children aged 0–12 years in Blantyre, Malawi, from 2013 to 2022 with CM as defined by the World Health Organization. Brain swelling on admission MRI was characterized by brain volume scores (BVS); severe swelling was scored as 7–8, mild-to-moderate as 4–6, normal as 3. The admission ONSD was measured via ultrasound; it was defined as abnormal if it was >4.5 mm in children >1 year and >4 mm in children

Spatially and temporally varying patterns of morphogen signals during development drive cell fate specification at the proper location and time. However, current in vitro methods typically do not allow for precise, dynamic spatiotemporal control of morphogen signaling and are thus insufficient to readily study how morphogen dynamics affect cell behavior. Here, we show that optogenetic Wnt/β-catenin pathway activation can be controlled at user-defined intensities, temporal sequences, and spatial patterns using engineered illumination devices for optogenetic photostimulation and light activation at variable amplitudes (LAVA). By patterning human embryonic stem cell (hESC) cultures with varying light intensities, LAVA devices enabled dose-responsive control of optoWnt activation and Brachyury expression. Furthermore, time-varying and spatially localized patterns of light revealed tissue patterning that models the embryonic presentation of Wnt signals in vitro. LAVA devices thus provide a low-cost, user-friendly method for high-throughput and spatiotemporal optogenetic control of cell signaling for applications in developmental and cell biology.