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Listeria monocytogenes is a bacterial pathogen that can cause life-threatening central nervous system (CNS) infections. While mechanisms by which L. monocytogenes and other pathogens traffic to the brain have been studied, a quantitative understanding of the underlying dynamics of colonization and replication within the brain is still lacking. In this study, we used barcoded L. monocytogenes to quantify the bottlenecks and dissemination patterns that lead to cerebral infection. Following intravenous (IV) inoculation, multiple independent invasion events seeded all parts of the CNS from the blood, however, only one clone usually became dominant in the brain. Sequential IV inoculations and intracranial inoculations suggested that clones that had a temporal advantage (i.e., seeded the CNS first), rather than a spatial advantage (i.e., invaded a particular brain region), were the main drivers of clonal dominance. In a foodborne model of cerebral infection with immunocompromised mice, rare invasion events instead led to a highly infected yet monoclonal CNS. This restrictive bottleneck likely arose from pathogen transit into the blood, rather than directly from the blood to the brain. Collectively, our findings provide a detailed quantitative understanding of the L. monocytogenes population dynamics that lead to CNS infection and a framework for studying the dynamics of other cerebral infections.

This mixed-methods study investigated the pandemic's impact on gifted identification among Arkansas's demographic groups and gifted services. The provision of gifted services during the pandemic varied, leading to differences in instructional practices and technology access. Nomination and identification processes were disrupted, hindering the recognition of giftedness, especially among underrepresented populations, and this was reflected in the identification numbers. Participants indicated that the pandemic has led to decreased funding for gifted programs, reduced administrator support, less time for students to participate in gifted classes, and increased responsibilities for gifted teachers. These factors have left teachers feeling overwhelmed and resulted in decreased student engagement, more behavioral issues, and academic struggles. The diversion of resources and priorities away from gifted education further exacerbated the inequities in gifted programs.

Abstract Microsatellite instability (MSI) occurs across a number of cancers and is associated with different clinical characteristics when compared to microsatellite stable (MSS) cancers. As MSI cancers have different characteristics, routine MSI testing is now recommended for a number of cancer types including colorectal cancer (CRC). Using gene panels for sequencing of known cancer mutations is routinely performed to guide treatment decisions. By adding a number of MSI regions to a small gene panel, the efficacy of simultaneous MSI detection in a series of CRCs was tested. Tumour DNA from formalin-fixed, paraffin-embedded (FFPE) tumours was sequenced using a 23-gene panel kit (ATOM-Seq) provided by GeneFirst. The mismatch repair (MMR) status was obtained for each patient from their routine pathology reports, and compared to MSI predictions from the sequencing data. By testing 29 microsatellite regions in 335 samples the MSI status was correctly classified in 314/319 samples (98.4% concordance), with sixteen failures. By reducing the number of regions in silico, comparable performance could be reached with as few as eight MSI marker positions. This test represents a quick, and accurate means of determining MSI status in FFPE CRC samples, as part of a routine gene mutation assay, and can easily be incorporated into a research or diagnostic setting. This could replace separate mutation and MSI tests with no loss of accuracy, thus improving testing efficiency.

Students and employees from Colorado State University School of Social Work partnered to co-lead a comprehensive student and employee community-building initiative focused on enhancing 2SLGBTQ+ student and employee well-being and belonging. The Queer Umbrella, created in response to queer and trans-identifying students and employees' experiences within a heterosexist and cissexist departmental climate, obtained funding from the university to improve departmental climate and 2SLGBTQ+ student and employee retention by focusing on three priority areas: visibility, action, and resilience. The ambitious semester-long grant initiative incorporated a variety of activities such as resiliency-based mutual aid groups and 2SLGBTQ+ alumnx lectures, advocacy, and queer and trans celebratory events. The purpose of this brief is to 1) share our experiences as leaders of the Queer Umbrella initiative, 2) highlight the impact of the initiative, and 3) identify and assess systemic barriers encountered while executing the initiative.

In the summer of 2020, SARS-CoV-2 infection rates among the U.S. population aged 20-39 years exceeded other age groups, with the largest increases occurring in the southern US. As many colleges reopened for in-person instruction in August and September, these trends continued among campuses across the country. Our study aimed to identify risk factors (demographic and behavioral) associated with SARS-CoV-2 infection among college students. We used data from a survey administered to students at a southern university in the US. The survey had a total of 765 respondents and this study included the 679 (88.8%) who responded about their SARS-CoV-2 infection status. We examined associations between population characteristics and reported SARS-CoV-2 infection and calculated prevalence ratios along with 95% confidence intervals. SARS-CoV-2 infection was 2.5 times more likely among current users of electronic nicotine delivery systems (ENDS) compared to those who do not use ENDS (95% confidence interval [CI]: (1.76-3.4)) and 2.8 times more likely among those who reported a high frequency of binge drinking compared to those who did not report binge drinking (95% CI: (1.81-4.36)). Current high frequency ENDS users were 2.76 (1.79-4.25) more likely to report SARS-CoV-2 infection than non-users. Current low frequency users of ENDS were 2.27 (1.53-3.37) times more likely to report SARS-CoV-2 infection than nonusers. A trend analysis among ENDS use frequency and SARS-CoV-2 infection was statistically significant, showing a significant dose response with increasing ENDS use. The results of this analysis may assist in providing guidance on policies as well as may serve as a steppingstone for future research concerning SAR-CoV-2 infection among university populations.

PURPOSE: The purpose of this study was to compare the histopathologic inflammation and fibrosis of orbital adipose tissue in orbital inflammatory disease (OID) specimens. METHODS: In this retrospective cohort study, inflammation, and fibrosis in orbital adipose tissue from patients with thyroid-associated orbitopathy (TAO), granulomatosis with polyangiitis (GPA), sarcoidosis, nonspecific orbital inflammation (NSOI), and healthy controls were scored by 2 masked ocular pathologists. Both categories were scored on a scale of 0 to 3 with scoring criteria based on the percentage of specimens containing inflammation or fibrosis, respectively. Tissue specimens were collected from oculoplastic surgeons at 8 international centers representing 4 countries. Seventy-four specimens were included: 25 with TAO, 6 with orbital GPA, 7 with orbital sarcoidosis, 24 with NSOI, and 12 healthy controls. RESULTS: The mean inflammation and fibrosis scores for healthy controls were 0.0 and 1.1, respectively. Orbital inflammatory disease groups inflammation (I) and fibrosis (F) scores, formatted [I, F] with respective p -values when compared to controls, were: TAO [0.2, 1.4] ( p = 1, 1), GPA [1.9, 2.6] ( p = 0.003, 0.009), sarcoidosis [2.4, 1.9] ( p = 0.001, 0.023), and NSOI [1.3, 1.8] ( p ≤ 0.001, 0.018). Sarcoidosis had the highest mean inflammation score. The pairwise analysis demonstrated that sarcoidosis had a significantly higher mean inflammation score than NSOI ( p = 0.036) and TAO ( p < 0.0001), but no difference when compared to GPA. GPA had the highest mean fibrosis score, with pairwise analysis demonstrating a significantly higher mean fibrosis score than TAO ( p = 0.048). CONCLUSIONS: Mean inflammation and fibrosis scores in TAO orbital adipose tissue samples did not differ from healthy controls. In contrast, the more intense inflammatory diseases such as GPA, sarcoidosis, and NSOI did demonstrate higher histopathologic inflammation and fibrosis. This has implications in prognosis, therapeutic selection, and response monitoring in orbital inflammatory disease.

BACKGROUND: In 2021, four patients who had received solid organ transplants in the USA developed encephalitis beginning 2-6 weeks after transplantation from a common organ donor. We describe an investigation into the cause of encephalitis in these patients. METHODS: From Nov 7, 2021, to Feb 24, 2022, we conducted a public health investigation involving 15 agencies and medical centres in the USA. We tested various specimens (blood, cerebrospinal fluid, intraocular fluid, serum, and tissues) from the organ donor and recipients by serology, RT-PCR, immunohistochemistry, metagenomic next-generation sequencing, and host gene expression, and conducted a traceback of blood transfusions received by the organ donor. FINDINGS: We identified one read from yellow fever virus in cerebrospinal fluid from the recipient of a kidney using metagenomic next-generation sequencing. Recent infection with yellow fever virus was confirmed in all four organ recipients by identification of yellow fever virus RNA consistent with the 17D vaccine strain in brain tissue from one recipient and seroconversion after transplantation in three recipients. Two patients recovered and two patients had no neurological recovery and died. 3 days before organ procurement, the organ donor received a blood transfusion from a donor who had received a yellow fever vaccine 6 days before blood donation. INTERPRETATION: This investigation substantiates the use of metagenomic next-generation sequencing for the broad-based detection of rare or unexpected pathogens. Health-care workers providing vaccinations should inform patients of the need to defer blood donation for at least 2 weeks after receiving a yellow fever vaccine. Despite mitigation strategies and safety interventions, a low risk of transfusion-transmitted infections remains. FUNDING: US Centers for Disease Control and Prevention (CDC), the Biomedical Advanced Research and Development Authority, and the CDC Epidemiology and Laboratory Capacity Cooperative Agreement for Infectious Diseases.

AbstractChronic pain affects more than 50 million Americans. Treatments remain inadequate, in large part, because the pathophysiological mechanisms underlying the development of chronic pain remain poorly understood. Pain biomarkers could potentially identify and measure biological pathways and phenotypical expressions that are altered by pain, provide insight into biological treatment targets, and help identify at-risk patients who might benefit from early intervention. Biomarkers are used to diagnose, track, and treat other diseases, but no validated clinical biomarkers exist yet for chronic pain. To address this problem, the National Institutes of Health Common Fund launched the Acute to Chronic Pain Signatures (A2CPS) program to evaluate candidate biomarkers, develop them into biosignatures, and discover novel biomarkers for chronification of pain after surgery. This article discusses candidate biomarkers identified by A2CPS for evaluation, including genomic, proteomic, metabolomic, lipidomic, neuroimaging, psychophysical, psychological, and behavioral measures. Acute to Chronic Pain Signatures will provide the most comprehensive investigation of biomarkers for the transition to chronic postsurgical pain undertaken to date. Data and analytic resources generatedby A2CPS will be shared with the scientific community in hopes that other investigators will extract valuable insights beyond A2CPS's initial findings. This article will review the identified biomarkers and rationale for including them, the current state of the science on biomarkers of the transition from acute to chronic pain, gaps in the literature, and how A2CPS will address these gaps.

Subnuclear compartmentalization has been proposed to play an important role in gene regulation by segregating active and inactive parts of the genome in distinct physical and biochemical environments. During X chromosome inactivation (XCI), the noncoding Xist RNA coats the X chromosome, triggers gene silencing and forms a dense body of heterochromatin from which the transcription machinery appears to be excluded. Phase separation has been proposed to be involved in XCI, and might explain the exclusion of the transcription machinery by preventing its diffusion into the Xist-coated territory. Here, using quantitative fluorescence microscopy and single-particle tracking, we show that RNA polymerase II (RNAPII) freely accesses the Xist territory during the initiation of XCI. Instead, the apparent depletion of RNAPII is due to the loss of its chromatin stably bound fraction. These findings indicate that initial exclusion of RNAPII from the inactive X reflects the absence of actively transcribing RNAPII, rather than a consequence of putative physical compartmentalization of the inactive X heterochromatin domain.

Following the global pandemic, educators relied heavily on live videoconferencing options and online meeting spaces to host class in lieu of traditional, in-person classroom learning. Yet, exhaustion and Zoom fatigue fueled a lack of engagement in such online spaces, while simultaneously the need for more informal connection to support learners' emotional well-being emerged. This study aims to better understand how online learners perceive the use of virtual reality (VR) as an alternative platform to informally connect and engage with one another, and to ascertain the impact on their satisfaction and motivation for such engagement. Specifically, the investigation sought to examine participant perceptions of social presence felt, the ability to connect and exchange informally, and the impact on motivation, digital literacy, and satisfaction overall.

Summary: Lineage transcription factors (TFs) provide one regulatory level of differentiation crucial for the generation and maintenance of healthy tissues. To probe TF function by measuring their dynamics during adult intestinal homeostasis, we established HILO-illumination-based live-cell single-molecule tracking (SMT) in mouse small intestinal enteroid monolayers recapitulating tissue differentiation hierarchies in vitro. To increase the throughput, capture cellular features, and correlate morphological characteristics with diffusion parameters, we developed an automated imaging and analysis pipeline, broadly applicable to two-dimensional culture systems. Studying two absorptive lineage-determining TFs, we found an expression level-independent contrasting diffusive behavior: while Hes1, key determinant of absorptive lineage commitment, displays a large cell-to-cell variability and an average fraction of DNA-bound molecules of ∼32%, Hnf4g, conferring enterocyte identity, exhibits more uniform dynamics and a bound fraction of ∼56%. Our results suggest that TF diffusive behavior could indicate the progression of differentiation and modulate early versus late differentiation within a lineage.

IntroductionInsomnia is associated with mild cognitive impairment, although the mechanisms of this impairment are not well-understood. Timing of slow-wave and rapid eye movement sleep may help explain cognitive impairments common in insomnia. This investigation aimed to determine whether cognitive changes following continuous theta-burst stimulation (cTBS) are attributable to active stimulation, polysomnographic parameters of sleep, or both.MethodData presented here are part of a pilot clinical trial aiming to treat insomnia by targeting a node in the default mode network using an inhibitory 40-s (cTBS). A double-blind counterbalanced sham-controlled crossover design was conducted. Participants (N = 20) served as their own controls on two separate in-laboratory visits—one with active cTBS and the other with sham cTBS. Each visit included cognitive assessments before and after stimulation and following a night of sleep in the lab monitored with polysomnography.ResultsSlow wave sleep duration influenced working memory in the active cTBS condition, with shorter duration predicting improvements in working memory post sleep (B = −0.003, p = 0.095). Onset latency to rapid eye movement sleep predicted subsequent working memory, regardless of treatment condition (B = −0.001, p = 0.040). Results suggest that changes in attention and processing speed were primarily due to slow wave sleep onset (B = −0.001, p = 0.017) and marginally predicted by slow wave sleep duration (B = 0.002, p = 0.081) and sleep efficiency (B = 0.006, p = 0.090).ConclusionsFindings emphasize the important role that timing of slow-wave and rapid eye movement sleep have on information processing. Future work using larger sample sizes and more stimulation sessions is needed to determine optimal interactions between timing and duration of slow wave and rapid eye movement throughout the sleep period.Clinical trial registrationThis study is registered on ClinicalTrials.gov (NCT04953559). https://clinicaltrials.gov/study/NCT04953559?locStr=Arizona&country=United%20States&state=Arizona&cond=insomnia&intr=tms%20&rank=1

Many employees are engaging in personal social media for work (PSMW), which involves posting work-related content from a user’s individual social-media account. Despite quantitative studies demonstrating the presence and outcomes of talking about work on social media, scholars know little about the process of using PSMW. To fill this gap, the current study uses social identity theory and boundary theory as conceptual frames to learn why and how people engage in PSMW. Through analyzing interview and observational data from employees’ social media across a variety of industries and work arrangements, we present a model of PSMW. Our findings contribute to scholarship in at least three ways. First, this study exposes the “work” behind PSMW through a pattern of Labored Worklife : a paradox of communicating both authentically and strategically. Second, our findings show PSMW as a distinctive, flexible way through which workers can traverse complicated work and non-work borders and communicate multiple (even conflicting) identities on social media. Third, this project suggests connections between PSMW and scholarship surrounding emotion at work.

Abstract Background Emerging adults (aged 18–29) are less likely to receive the COVID-19 vaccine than any other adult age group. Black Americans are less likely than non-Hispanic white Americans to be fully vaccinated against COVID-19. This study explored factors which affect vaccine intention and attitudes in Black American emerging adults with asthma. Methods Participants were recruited from an NHLBI-funded clinical trial to improve asthma control. Fifty-nine Black American emerging adults completed a Qualtrics survey that assessed asthma control, intention to vaccinate, and factors which may affect the decision to vaccinate. Twenty-five participants also completed a semi-structured interview via Zoom. Bivariate correlations and descriptive statistics, including Chi Square analyses, were run using SPSS. Interview thematic analyses were conducted via QDA Miner. Results Of the 59 Black American emerging adults with asthma who completed surveys, 32.2% responded that they were highly unlikely to receive the COVID-19 vaccine, while 50.8% responded that they were highly likely to receive it. Increased asthma control was significantly correlated with a higher likelihood to discuss the COVID-19 vaccine with their healthcare provider (ρ = 0.339, α = 0.011). Concerns about immediate (ρ= -0.261, α = 0.050) and long-term (ρ= -0.280, α = 0.035) side effects were inversely correlated with intention to vaccinate. Only 17% of the participants who were unemployed stated that they were highly likely to receive the vaccines compared to 65% of the participants who were employed; however, interview participants who were unemployed stated not needing the vaccine because they were protecting themselves by social distancing. When deciding whether to receive the vaccine, safety, efficacy, and immediate side effects were the top three factors for 91%, 54%, and 49% of the participants, respectively. Beliefs about the vaccines’ safety and efficacy, information gathering, personal factors, and societal factors emerged as important themes from the interviews. Conclusion Only half of the surveyed Black American emerging adults with asthma were highly likely to receive the COVID-19 vaccine. Safety and efficacy were important for the majority of the participants, regardless of vaccine intention. Greater asthma control, but not access to asthma-related healthcare, was correlated with intention to discuss the vaccine with their healthcare provider.

Abstract Background Low back pain (LBP) is a significant public health problem, is very prevalent, and is often characterized by the persistence of symptoms. Transcutaneous electrical nerve stimulation (TENS) may benefit people with chronic LBP because it can activate descending inhibitory pathways and inhibit central excitability. However, previous studies that have investigated the effects of TENS on pain in people with LBP have failed to use proper intensities of current, and the timing of the assessment of pain was not performed during the peak of the analgesic response or functional activities. Therefore, the present study aims to assess the effects of TENS on measures of pain, function, and descending inhibition using the maximal tolerable intensity of TENS in participants with LBP. Methods/design This study will be a randomized crossover trial. The participants for this study will be recruited from various places, including the University of Hartford, physical therapy clinics, and local businesses in the Hartford area, as well as online websites geared towards clinical trial recruitment. A total of 34 participants will receive all three treatments: active TENS, placebo TENS, and no treatment control. The treatment order will be randomized using a website-based randomization tool. For active TENS, a modulating frequency of 2–125 Hz will be applied with a variable pulse duration and maximal tolerable intensity for 30 min. The TENS will be left on for post-treatment testing to assess the effects during its maximally effective period for a total of 50 to 60 min. Furthermore, the intensity may be turned down if muscle twitching is present to ensure blinding of the evaluator. For placebo TENS, the unit will deliver current for 45 s, ramping to 0 in the last 15 s. The primary outcome will be pain intensity at rest and with movement, determined using the numerical pain rating scale. The secondary outcomes will be pressure pain threshold, heat pain threshold, temporal summation of pain, conditioned pain modulation, sit-to-stand test, and repeated trunk flexion. The assessments will be performed immediately before and after treatment. Statistical analysis of the data obtained will consider a significance level of p

Abstract Wildland fires contribute significantly to the ambient air pollution burden worldwide, causing a range of adverse health effects in exposed populations. The toxicity of woodsmoke, a complex mixture of gases, volatile organic compounds, and particulate matter, is commonly studied in vitro using isolated exposures of conventionally cultured lung cells to either resuspended particulate matter or organic solvent extracts of smoke, leading to incomplete toxicity evaluations. This study aimed to improve our understanding of the effects of woodsmoke inhalation by building an advanced in vitro exposure system that emulates human exposure of the airway epithelium. We report the development and characterization of an innovative system that permits live-cell monitoring of the intracellular redox status of differentiated primary human bronchial epithelial cells cultured at an air–liquid interface (pHBEC-ALI) as they are exposed to unfractionated woodsmoke generated in a tube furnace in real time. pHBEC-ALI exposed to freshly generated woodsmoke showed oxidative changes that were dose-dependent and reversible, and not attributable to carbon monoxide exposure. These findings show the utility of this novel system for studying the molecular initiating events underlying woodsmoke-induced toxicity in a physiologically relevant in vitro model, and its potential to provide biological plausibility for risk assessment and public health measures.

Mulberry Diels-Alder-type adducts (MDAAs), isolated from Morus alba root bark, exhibit dual activity against viral and bacterial pathogens but show sobering efficacy following oral administration. Inhalation administration may overcome issues with oral bioavailability and improve efficacy for the treatment of respiratory infections. To assess the suitability of MDAAs for inhalation administration, physicochemical (e.g. pH, pKa, logP, pH-dependent solubility) and biopharmaceutical (epithelial cytotoxicity, permeability, and uptake) properties of two bioactive MDAA stereoisomers sanggenon C (SGC) and sanggenon D (SGD) were evaluated as isolated natural compounds and within parent extracts (MA21, MA60). Despite their structural similarity, SGD exhibited a 10-fold higher solubility than SGC across pH 1.2–7.4, with slight increases at neutral pH. Both compounds were more soluble in isolated form than in the parent extracts. The more lipophilic SGC was found to be more cytotoxic when compared to SGD, indicating a better cellular penetration, which was confirmed by uptake studies. Nonetheless, SGC and SGD exhibited no measurable permeability across intact Calu-3 monolayers, highlighting their potential for increased lung retention and improved local anti-infective activity following inhalation administration. Results suggest that SGC and SGD in isolated form, rather than as extracts, are promising candidates for pulmonary drug delivery to treat lung infections.

Mycotoxin exposure during pregnancy has been associated with adverse birth outcomes and poor infant growth in low- and middle-income countries. We assessed multiple biomarkers and metabolites of exposure to mycotoxins during pregnancy and their associations with birth outcomes and infant growth in 305 pregnant participants, between 30 and 34 completed weeks of gestation, in rural Burkina Faso. In this study, whole blood microsamples were analyzed for mycotoxin concentrations using ultra-performance liquid chromatography coupled to tandem mass spectrometry. Unadjusted and adjusted associations between mycotoxin exposure, and birth outcomes and infant growth at 6 months were estimated using linear regression models for continuous outcomes and linear probability models with robust variance estimation for binary outcomes. Infant growth trajectories from birth to 6 months were compared by exposure status using mixed-effects models with random intercept for the individual infant and random slope for the infant's age. Ochratoxin A (OTA) exposure was detected in 50.8 % of the study participants, with aflatoxin G1, aflatoxin M1, cyclopiazonic acid, deoxynivalenol and T-2-toxin being detected in the range of 0.33 % and 2.31 % of the population. We found no statistically significant (p ≥ 0.05) associations between OTA exposure, and birth outcomes and infant growth. Despite this, the findings indicate a significant presence of ochratoxin A among pregnant participants. Public health policies and nutrition-sensitive interventions must ensure that OTA exposure is reduced in Burkina Faso.

ABSTRACTER+ breast cancers (BC) are characterized by the elevated expression and signaling of estrogen receptor alpha (ESR1), which renders them sensitive to anti-endocrine therapy. While these therapies are clinically effective, prolonged treatment inevitably results in therapeutic resistance, which can occur through the emergence of gain-of-function mutations in ESR1. The central importance of ESR1 and development of mutated forms of ESR1 suggest that vaccines targeting these proteins could potentially be effective in preventing or treating endocrine resistance. To explore the potential of this approach, we developed several recombinant vaccines encoding different mutant forms of ESR1 (ESR1mut) and validated their ability to elicit ESR1-specific T cell responses. We then developed novel ESR1mut-expressing murine mammary cancer models to test the anti-tumor potential of ESR1mut vaccines. We found that these vaccines could suppress tumor growth, ESR1mut expression and estrogen signaling in vivo. To illustrate the applicability of these findings, we utilize HPLC to demonstrate the presentation of ESR1 and ESR1mut peptides on human ER+ BC cell MHC complexes. We then show the presence of human T cells reactive to ESR1mut epitopes in an ER+ BC patient. These findings support the development of ESR1mut vaccines, which we are testing in a Phase I clinical trial.

Heme is an essential cofactor for multiple cellular processes in most organisms. In developing erythroid cells, the demand for heme synthesis is high, but is significantly lower in non-erythroid cells. While the biosynthesis of heme in metazoans is well understood, the tissue-specific regulation of the pathway is less explored. To better understand this, we analyzed the mitochondrial heme metabolon in erythroid and non-erythroid cell lines from the perspective of ferrochelatase (FECH), the terminal enzyme in the heme biosynthetic pathway. Affinity purification of FLAG-tagged-FECH, together with mass spectrometric analysis, was carried out to identify putative protein partners in human and murine cell lines. Proteins involved in the heme biosynthetic process and mitochondrial organization were identified as the core components of the FECH interactome. Interestingly, in non-erythroid cell lines, the FECH interactome is highly enriched with proteins associated with the tricarboxylic acid (TCA) cycle. Overall, our study shows that the mitochondrial heme metabolon in erythroid and non-erythroid cells has similarities and differences, and suggests new roles for the mitochondrial heme metabolon and heme in regulating metabolic flux and key cellular processes.

We investigated wind-wave and suspended-sediment dynamics in Little Holland Tract and Liberty Island, two subsided former agricultural tracts in the Cache Slough complex in the northern Sacramento-San Joaquin Delta which were restored to tidal shallows to improve habitat. Turbidity, and thus suspended-sediment concentration (SSC), is important to habitat quality because some species of native fishes, including the Delta Smelt, are found preferentially in more turbid waters. Data from October 2015 to August 2016 show that average SSC was greater within Little Holland Tract than in the primary breach that connects the basin to surrounding channels: approximately twice as great at a shallower station farther from the breach and 15% greater at a deeper station closer to the breach. Suspended-sediment concentration within Little Holland Tract was directly related to wave shear stress and inversely related to water depth, based on linear regression. We used measurements of suspended-sediment flux (SSF) through the largest levee breaches to assess whether the enhanced SSC within Little Holland Tract is exported to surrounding waters, thus potentially increasing turbidity over a wider region. Cumulatively, sediment is exported through the Little Holland Tract breaches in winter and imported in summer, consistent with regional patterns in sediment flux, indicating that wind-wave re-suspension within the basin does not control sediment flux from Little Holland Tract on seasonal time-scales. Some sediment was exported during wind-wave events, and results show that sediment export is greater when primary breaches are located downwind of the basin rather than upwind.

The 2020-2021 academic year brought numerous challenges to teachers across the country as they worked to educate students amidst the COVID-19 pandemic. The current study is a secondary data analysis of qualitative responses collected as part of a teacher survey to evaluate a social emotional learning curriculum implemented during the 2020-2021 academic year. The lived experiences of teachers (N = 52) across 11 elementary schools in the Great Plains region were captured through open-ended questions as the teachers transitioned from in-person to remote learning. A phenomenological approach was utilized to analyze the challenges expressed by teachers as they faced instability and additional professional demands. Given that stress and other factors that strain mental health exist within multiple layers of an individual's social ecology, a modified social-ecological framework was used to organize the results and themes. Findings suggest that during the academic year, teachers experienced stressors related to their personal and professional roles, concerns for students' well-being which extended beyond academics, and frustrations with administration and other institutional entities around COVID safety measures. Without adequate support and inclusion of teacher perspectives, job-related stress may lead to teacher shortages, deterioration of teacher mental health, and ultimately worse outcomes for students. Implications for policy, research, and practice are discussed.

Background: Gram-negative bacilli (GNB) bacteremia is a common and potentially fatal infection with mortality rates estimated to be 14-34%, despite effective antimicrobial treatments. Follow-up blood cultures (FUBCs) are blood cultures that are repeated after an initial positive culture and are indicated in certain clinical scenarios, such as in candidemia or Staphylococcus aureus bacteremia to ensure clearance of the bloodstream infection. FUBCs are used in bacteremia to assess the appropriateness and duration of antimicrobial therapy. Currently, there are no guidelines in place regarding the use of FUBC for GNB bacteremia. Furthermore, the utility of FUBCs is not well-studied in adults living with cancer. The purpose of this study is to identify risk factors associated with persistent GNB bacteremia in adult patients with cancer. Methods: We conducted a single-center, retrospective study in patients aged ≥18 years, hospitalized during calendar year 2022, living with cancer, and with laboratory confirmed GNB bloodstream infection. FUBC was defined as a blood culture performed within 7 days of the initial positive blood culture. Patients were grouped as having the same organism on FUBC (+/same), FUBC with no growth (NG) (+/NG), and different organism on FUBC (+/Different). Patients with a different organism on FUBC were excluded from analysis. We gathered demographic information, suspected source of bacteremia, type of malignancy, identified organisms, presence of antimicrobial resistance, and comorbidities (eg. presence of central venous catheters, urinary catheters, end-stage renal disease). Categorical variables were compared using Chi-square or Fisher’s exact test. Continuous variables were compared using Wilcoxon rank sum tests. Logistic regression analysis was used to identify the independent predictors of persistent GNB bacteremia. Results: 356 unique patients with FUBC were identified after inclusion/exclusion criteria. 93/356 (26%) of patients had persistent GNB bacteremia (+/same). Multivariate analysis identified history of bacteremia within the preceding year (OR 2.95, 95% CI [1.6-5.6]) and Achromobacter spp. bacteremia (OR 10.03, 95% CI [1.59-63.23] as independent risk factors for persistence. Organisms with multidrug resistance such as extended-spectrum beta-lactamase (OR 2.47, 95% CI [1.21-5.07]) and carbapenem-resistant organisms (OR 3.35, 95% CI [1.04-10.81]) were also associated with persistent GNB bacteremia. Conclusions: This is the first study to specifically identify risk factors for persistent GNB bacteremia in patients living with cancer. FUBC may be useful in GNB bacteremia with less common organisms and/or if they exhibit multidrug resistance on susceptibility testing. The utility of FUBC should be further explored in patients with cancer with certain risk factors.

IntroductionMRI-guided radiotherapy (MRgRT) allows for direct motion management and real-time radiation treatment plan adaptation. We report our institutional experience using low strength 0.35T MRgRT for thoracic malignancies, and evaluate changes in treatment duty cycle between first and final MRgRT fractions.MethodsAll patients with intrathoracic tumors treated with MRgRT were included. The primary reason for MRgRT (adjacent organ at risk [OAR] vs. motion management [MM] vs. other) was recorded. Tumor location was classified as central (within 2cm of tracheobronchial tree) vs. non-central, and further classified by the Expanded HILUS grouping. Gross tumor volume (GTV) motion, planning target volume expansions, dose/fractionation, treatment plan time, and total delivery time were extracted from the treatment planning system. Treatment plan time was defined as the time for beam delivery, including multileaf collimator (MLC) motion, and gantry rotation. Treatment delivery time was defined as the time from beam on to completion of treatment, including treatment plan time and patient respiratory breath holds. Duty cycle was calculated as treatment plan time/treatment delivery time. Duty cycles were compared between first and final fraction using a two-sample t-test.ResultsTwenty-seven patients with thoracic tumors (16 non-small cell lung cancer and 11 thoracic metastases) were treated with MRgRT between 12/2021 and 06/2023. Fifteen patients received MRgRT due to OAR and 11 patients received MRgRT for motion management. 11 patients had central tumors and all were treated with MRgRT due to OAR risk. The median dose/fractionation was 50 Gy/5 fractions. For patients treated due to OAR (n=15), 80% had at least 1 adapted fraction during their course of radiotherapy. There was no plan adaptation for patients treated due to motion management (n=11). Mean GTV motion was significantly higher for patients treated due to motion management compared to OAR (16.1mm vs. 6.5mm, p=0.011). Mean duty cycle for fraction 1 was 54.2% compared to 62.1% with final fraction (p=0.004). Mean fraction 1 duty cycle was higher for patients treated due to OAR compared to patients treated for MM (61% vs. 45.0%, p=0.012).DiscussionDuty cycle improved from first fraction to final fraction possibly due to patient familiarity with treatment. Duty cycle was improved for patients treated due to OAR risk, likely due to more central location and thus decreased target motion.

Background: Patients discharged to non-home facilities (NHD) after total hip arthroplasty (THA) and total knee (TKA) arthroplasty experience higher rates of adverse events and may require more aggressive venous thromboembolism (VTE) chemoprophylaxis. Our aim was to compare the rates of VTE in NHD patients and those discharged home (HD) after THA/TKA. Our secondary aim was to determine VTE rates within HD and NHD groups when stratified by chemoprophylactic regimen. Methods: A retrospective cohort of primary THA and TKA patients were stratified into HD and NHD, then allocated into groups by chemoprophylactic regimen on discharge: aspirin alone (AA), more aggressive (MA) chemoprophylaxis, and other regimens (other). The primary outcome was VTE. Rates of VTE in HD and NHD patients, as well as AA and MA regimens, were analyzed using a generalized linear regression model. Results: Six thousand three hundred seventy-nine patients were included with 1.03% experiencing VTE. HD had lower rates of VTE compared to NHD (0.83% vs 2.17%, P < .001). AA had similar rates of VTE compared to MA (0.99% vs 1.08%, P = .82). NHD patients had a lower VTE rate with MA vs AA prophylaxis (1.47% vs 3.83%, P = .016). HD patients treated with AA vs MA had no difference in VTE rates (0.76% vs 0.96%, P = .761). Conclusions: NHD patients have higher rates of VTE than HD patients. However, NHD patients have significantly lower rates of VTE on MA chemoprophylaxis compared to those on AA. Providers should consider prescribing MA VTE chemoprophylaxis for NHD patients. Prospective, randomized studies are necessary to confirm these recommendations.

Nature utilizes three distinct pathways to synthesize the essential enzyme cofactor heme. The coproporphyrin III-dependent pathway, predominantly present in Bacillaceae, employs an oxygen-dependent coproporphyrinogen III oxidase (CgoX) that converts coproporphyrinogen III into coproporphyrin III. In this study, we report the bioinformatic-based identification of a gene called ytpQ, encoding a putative oxygen-independent counterpart, which we propose to term CgoN, from Priestia (Bacillus) megaterium. The recombinantly produced, purified, and monomeric YtpQ (CgoN) protein is shown to catalyze the oxygen-independent conversion of coproporphyrinogen III into coproporphyrin III. Minimal non-enzymatic conversion of coproporphyrinogen III was observed under the anaerobic test conditions employed in this study. FAD was identified as a cofactor, and menadione served as an artificial acceptor for the six abstracted electrons, with a KM value of 3.95 μmol/L and a kcat of 0.63 per min for the substrate. The resulting coproporphyrin III, in turn, acts as an effective substrate for the subsequent enzyme of the pathway, the coproporphyrin III ferrochelatase (CpfC). Under aerobic conditions, oxygen directly serves as an electron acceptor, but is replaced by the more efficient action of menadione. An AlphaFold2 model of the enzyme suggests that YtpQ adopts a compact triangular shape consisting of three domains. The N-terminal domain appears to be flexible with respect to the rest of the structure, potentially creating a ligand binding site that opens and closes during the catalytic cycle. A catalytic mechanism similar to the oxygen-independent protoporphyrinogen IX oxidase PgoH1 (HemG), based on the flavin-dependent abstraction of six electrons from coproporphyrinogen III and their potential quinone-dependent transfer to a membrane-localized electron transport chain, is proposed.

Since the 1970s, international students have represented a growing proportion of the global student body, yet how they adjust and how universities can support them is relatively unexplored. We conducted a qualitative study of 36 international students of 11 nationalities studying in Norway and the UK and found that their experience did not fit the dominant 'U-Curve' of adaptation that suggests there is a honeymoon period on arrival. Confirmed with conversations with student wellbeing staff, who suggest that anxiety and culture shock are the norm, the data allows us to suggest factors that trigger adjustment and interventions to improve students' experience. Our contribution is a 'J-Curve' model comprising cultural challenge, adjustment, and mastery, to reflect the reality of the international student experience.

Abstract Background Equitable sex- and gender-based representation in clinical trials is an essential step to ensuring evidence-based care for women. While multi-institutional actions have led to significant improvements in the inclusion of women in trials, inequity persists in areas like sex-neutral cancers and cardiovascular disease. We sought to identify strategies described or evaluated to boost the inclusion of women in clinical trials. Methods We used evidence mapping methodology to examine the breadth of relevant literature. We developed an a priori protocol and followed reporting guidance from the Preferred Reporting Items for Systematic Reviews and Meta-Analysis where applicable. We searched MEDLINE® (via PubMed) and EMBASE (via Elsevier) databases from inception through April 4, 2023, and used standardized procedures incorporating duplication and data verification. We included articles that described strategies to improve the recruitment and retention of women in clinical trials. Results We identified 122 articles describing recruitment and retention strategies for 136 trials (377,595 women). Only one article distinguished between the sex and gender identity of participants, and none defined their use of the terms such as “women” or “female”. The majority of articles (95%) described recruitment for only women, and 64% were conducted in the USA. Ninety-two articles (75%) described strategies in the context of sex-specific conditions (e.g., gynecologic diagnosis). The majority of included articles evaluated a behavioral intervention (52%), with 23% evaluating pharmacologic interventions and 4% invasive interventions. The most common trial phase for reported strategies was during outreach to potential participants (116 articles), followed by intervention delivery (76), enrollment (40), outcomes assessment (21), analysis and interpretation (3), and dissemination (4). We describe specific types of strategies within each of these phases. Conclusions Most of the existing literature describing strategies to improve the inclusion of women draws from trials for sex-specific conditions and is largely related to outreach to potential participants. There is little information about how and if studies have attempted to proportionally increase the inclusion of women in trials with both men and women or those focused on invasive and pharmacologic interventions. Future work in this area should focus on how to increase the participation of women in mixed-sex studies and on those areas with remaining inequities in trial participation.