Emergence of antibiotic-specific Mycobacterium tuberculosis phenotypes during prolonged treatment of mice.

A major challenge in tuberculosis (TB) therapeutics is that antibiotic exposure leads to changes in the physiologic state of M. tuberculosis (Mtb) which may enable the pathogen to withstand treatment. While antibiotic-treated Mtb have been evaluated in short-term in vitro experiments, it is unclear if and how long-term in vivo treatment with diverse antibiotics with varying treatment-shortening activity (sterilizing activity) affect Mtb physiologic states differently. Here, we used SEARCH-TB, a pathogen-targeted RNA-sequencing platform, to characterize the Mtb transcriptome in the BALB/c high-dose aerosol infection mouse model following 4-week treatment with three sterilizing and three non-sterilizing antibiotics. Certain transcriptional changes were concordant among most antibiotics, including decreased expression of genes associated with protein synthesis and metabolism, and the induction of certain genes associated with stress responses. However, the magnitude of this concordant response differed between antibiotics. Sterilizing antibiotics rifampin, pyrazinamide, and bedaquiline generated a more quiescent Mtb state than did non-sterilizing antibiotics isoniazid, ethambutol, and streptomycin, as indicated by decreased expression of genes associated with translation, transcription, secretion of immunogenic proteins, metabolism, and cell wall synthesis. Additionally, we identified distinguishing transcriptional effects specific to each antibiotic, indicating that different mechanisms of action induce distinct patterns of cellular injury. In addition to elucidating Mtb physiologic changes associated with antibiotic stress, this study demonstrates the value of SEARCH-TB as a highly granular pharmacodynamic assay that reveals antibiotic effects that are not apparent based on culture alone.

Toxoplasma FER1 is a versatile and dynamic mediator of differential microneme trafficking and microneme exocytosis.

Toxoplasma gondii is a polarized cell concentrating several secretory organelles at the apical pole. The secretory micronemes come in two sub-populations differentiated by dependence on Rab5A/C in their biogenesis. Calcium-dependent exocytosis of micronemes occurs at the very apical tip and is critical for parasite egress from its host cell, adhesion and invasion of the next cell. Ferlins represent a protein family with roles in exocytosis containing multiple Ca2+-sensing C2 domains. We determined that T. gondii's ferlin 1 (FER1) localized dynamically to the parasite's secretory pathway. FER1 function was dissected by dominant negative overexpression strategies. We demonstrated that FER1 traffics microneme organelles along the following trajectories: (1) Along the cortex to the apical end; (2) To the apical tip for fusion with the plasma membrane; (3) Differential microneme sub-population traffic, and that FER1 could putatively be responsible for microneme protein trafficking. (4) From the trans-Golgi-endosomal network to the subpellicular cortex; (5) Retrograde transport allowing microneme recycling from mother to daughter. Finally, FER1 overexpression triggers a microneme exocytosis burst, supporting the notion that the radially organized micronemes at the apical tip comprise a readily-releasable microneme pool. In summary, FER1 is pivotal for dynamic microneme trafficking, acts differently on the two microneme subpopulations, and acts on the plasma membrane fusion step during microneme exocytosis.

"I was very scared when I found out my son had sickle cell": Caregiver knowledge and attitudes toward early intervention for young children with sickle cell disease: Implications for policy and practice from a multi-site study.

OBJECTIVE: This study characterized caregivers' beliefs related to early intervention services for children with sickle cell disease (SCD) to gain an indepth understanding of caregivers' experiences and desires for early intervention services. METHODS: Both qualitative and quantitative data were collected from caregivers of children aged 0-4 years with SCD across two sites in the United States. Caregivers completed the Knowledge of Infant Development Inventory, a custom survey about their experiences with early intervention, and a qualitative interview. RESULTS: A total of 127 caregivers were approached, 47 participated in surveys, and 20 completed interviews. Caregivers expressed varying levels of confidence and understanding of developmental milestones across sites. Interviews highlighted three main themes: fear of SCD-related complications, variable buy-in to early intervention, and the importance of provider-caregiver relationships. While some caregivers appreciated early intervention, others questioned its necessity. Caregivers communicated interest in connecting with other families facing similar challenges, emphasizing the need for increased awareness of available resources. CONCLUSIONS: Fear about their child's well-being was expressed by many caregivers, emphasizing the need for a supportive healthcare team that can help families connect with preventive interventions. While about a quarter of children had been referred to rehabilitation services, caregivers were unaware of the elevated risk for developmental delay, which diminished caregiver interest in participating in programs like early intervention. This study underscores the importance of addressing knowledge gaps and overcoming barriers to enhance care for families affected by SCD.

The association of maintenance hormone therapy with overall survival in advanced-stage low-grade serous ovarian carcinoma: A risk-set matched retrospective study.

OBJECTIVE: We conducted a multi-institutional observational study to investigate whether maintenance hormone therapy following primary treatment of low-grade advanced-stage ovarian cancer (LGSOC) is associated with an overall survival advantage. METHODS: We included patients with histologically confirmed stage III or IV LGSOC diagnosed between Jan 1, 2004, and Dec 31, 2019, treated in Commission on Cancer-accredited cancer programs in the US. Patients who received hormone therapy within six months of diagnosis were matched to controls who did not initiate hormone therapy during this timeframe by risk-set propensity score matching. The primary outcome was the risk of death from any cause within five years of initiation of HT or observation. RESULTS: There were 296 patients who initiated maintenance hormone therapy within six months of diagnosis and 2805 potential controls. Patients who received hormone therapy were more often treated in academic medical centers (55% vs. 44%), diagnosed later in the study period (62% vs. 23% diagnosed in 2018-2019), and frequently received no chemotherapy during initial treatment (45% vs. 17%). After risk set propensity score matching, we identified 225 patients treated with HT and 225 untreated controls who were otherwise similar with respect to measured covariates. In the matched cohort, hormone therapy was associated with a reduction in the risk of death (hazard ratio 0.60; 95% CI 0.38-0.94), corresponding to a 60-month survival of 75% compared with 65%. CONCLUSIONS: Following primary management of LGSOC, maintenance hormone therapy was associated with improved overall survival compared with observation.

Comparing upfront catheter-based thrombectomy with alternative treatment strategies for clot-in-transit.

Clot-in-transit (CIT) is associated with high mortality, and optimal treatment strategies remain uncertain. This study compares the efficacy of catheter-based thrombectomy (CBT) with other treatments for CIT, including anticoagulation, systemic thrombolytic (ST) therapy, and surgical thrombectomy. We conducted a retrospective analysis of patients with CIT documented on echocardiography between January 2020 and May 2024, managed with urgent upfront CBT. We compared the all-cause mortality rates of the CBT cohort to performance goal rates for anticoagulation, systemic thrombolysis (ST), and surgical thrombectomy from a published meta-analysis. Our cohort included 26 patients who underwent CBT (mean age 59.3 ± 17.9 years, 42.3% women, 57.7% Black). Compared to 463 patients from the meta-analysis receiving alternative treatments, the CBT group's short-term mortality was significantly lower (7.7% vs 32.4% for anticoagulation, 13.8% for ST, and 23.2% for surgical thrombectomy). CBT demonstrated noninferiority to anticoagulation (P < .001), ST (P = .031) and surgical thrombectomy (P < .001), and was superior to anticoagulation (P = .0056) and surgical thrombectomy (P = .036). This study suggests CBT is a promising treatment for CIT. Further prospective studies are warranted to validate these findings.

Differences in Self-Reported Stress Versus. Hair and Nail Cortisol Among Adolescent and Young Adult Males.

BACKGROUND: Chronic stress causes harmful physiologic responses that yield increased inflammation and subsequent health conditions. Stress is an important measure among minoritized populations who face social situations that predispose risk to developing mental health problems. Hair and fingernail cortisol have been studied as retrospective measures of chronic stress and to demonstrate biological response to social situations. OBJECTIVES: The objective of this study was to compare the Perceived Stress Scale (PSS) with hair and nail cortisol concentrations and assess the risk factors associated with stress levels among heterosexual and sexual and gender-minoritized adolescent males. METHODS: We recruited a cohort of adolescents who were assigned male sex at birth. Approximately half of our cohort consisted of sexual and gender-minoritized people, and half consisted of heterosexual cisgender males. Participants provided hair and nail samples and completed a survey that included demographic and hair hygiene questions, and the PSS. Hair and nail samples were processed in a lab and survey results were analyzed descriptively. RESULTS: Several samples were not provided or received, and some survey data was missing. Hair and nail cortisol values were significantly correlated. There was no significant relationship between the PSS and hair and nail cortisol values. No significant differences were found between the heterosexual and sexual minoritized groups. Black participants reported lower perceived stress scores compared to White participants. Participants whose gender was non-binary or genderqueer had higher hair cortisol values compared to those who identified as male. Older participants had higher hair cortisol values compared to younger participants. DISCUSSION: Previous researchers have similarly found no correlation between self-report stress scales and cortisol values, increased stress experience among non-binary or genderqueer individuals compared to cisgender individuals, and a positive correlation between aging and stress. Yet, our finding that Black participants reported lower stress levels than White participants is unexpected. Our study demonstrates a high correlation between hair and nail cortisol values, suggeting the potential to interchange these markers as needed.

Optimizing Response Rates to Examine Health IT Maturity and Nurse Practitioner Care Environments in US Nursing Homes: Mixed Mode Survey Recruitment Protocol.

BACKGROUND: Survey-driven research is a reliable method for large-scale data collection. Investigators incorporating mixed-mode survey designs report benefits for survey research including greater engagement, improved survey access, and higher response rate. Mix-mode survey designs combine 2 or more modes for data collection including web, phone, face-to-face, and mail. Types of mixed-mode survey designs include simultaneous (ie, concurrent), sequential, delayed concurrent, and adaptive. This paper describes a research protocol using mixed-mode survey designs to explore health IT (HIT) maturity and care environments reported by administrators and nurse practitioners (NPs), respectively, in US nursing homes (NHs). OBJECTIVE: The aim of this study is to describe a research protocol using mixed-mode survey designs in research using 2 survey tools to explore HIT maturity and NP care environments in US NHs. METHODS: We are conducting a national survey of 1400 NH administrators and NPs. Two data sets (ie, Care Compare and IQVIA) were used to identify eligible facilities at random. The protocol incorporates 2 surveys to explore how HIT maturity (survey 1 collected by administrators) impacts care environments where NPs work (survey 2 collected by NPs). Higher HIT maturity collected by administrators indicates greater IT capabilities, use, and integration in resident care, clinical support, and administrative activities. The NP care environment survey measures relationships, independent practice, resource availability, and visibility. The research team conducted 3 iterative focus groups, including 14 clinicians (NP and NH experts) and recruiters from 2 national survey teams experienced with these populations to achieve consensus on which mixed-mode designs to use. During focus groups we identified the pros and cons of using mixed-mode designs in these settings. We determined that 2 mixed-mode designs with regular follow-up calls (Delayed Concurrent Mode and Sequential Mode) is effective for recruiting NH administrators while a concurrent mixed-mode design is best to recruit NPs. RESULTS: Participant recruitment for the project began in June 2023. As of April 22, 2024, a total of 98 HIT maturity surveys and 81 NP surveys have been returned. Recruitment of NH administrators and NPs is anticipated through July 2025. About 71% of the HIT maturity surveys have been submitted using the electronic link and 23% were submitted after a QR code was sent to the administrator. Approximately 95% of the NP surveys were returned with electronic survey links. CONCLUSIONS: Pros of mixed-mode designs for NH research identified by the team were that delayed concurrent, concurrent, and sequential mixed-mode methods of delivering surveys to potential participants save on recruitment time compared to single mode delivery methods. One disadvantage of single-mode strategies is decreased versatility and adaptability to different organizational capabilities (eg, access to email and firewalls), which could reduce response rates. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/56170.

Evaluating the impact of a year-long external mentorship pilot program in classical hematology.

ABSTRACT: Effective mentorship is a pivotal factor in shaping the career trajectory of trainees interested in classical hematology (CH), which is of critical importance due to the anticipated decline in the CH workforce. However, there is a lack of mentorship opportunities within CH compared with medical oncology. To address this need, a year-long external mentorship program was implemented through the American Society of Hematology Medical Educators Institute. Thirty-five hematology/oncology fellows interested in CH and 34 academically productive faculty mentors from different institutions across North America were paired in a meticulous process that considered individual interests, experiences, and background. Pairs were expected to meet virtually once a month. Participation in a scholarly project was optional. A mixed-methods sequential explanatory design was used to evaluate the program using mentee and mentor surveys, a mentee interview, and a mentee focus group. Thirty-three mentee-mentor pairs (94.2%) completed the program. Sixty-three percent of mentee respondents worked on a scholarly project with their mentor; several mentees earned publications, grants, and awards. Mentee perception that their assigned mentor was a good match was associated with a perceived positive impact on confidence (P = .0423), career development (P = .0423), and professional identity (P = .0302). Furthermore, 23 mentees (66%) accepted CH faculty positions after fellowship. All mentor respondents believed that this program would increase retention in CH. This mentorship program demonstrates a productive, beneficial way of connecting mentees and mentors from different institutions to improve the careers of CH trainees, with the ultimate goal of increasing retention in CH.

Decreased ability to manage increases in reactive oxygen species may underlie susceptibility to arrhythmias in mice lacking Scn1b.

Scn1b plays essential roles in the heart, where it encodes ß1-subunits that serve as modifiers of gene expression, cell surface channel activity, and cardiac conductivity. Reduced ß1 function is linked to electrical instability in various diseases with cardiac manifestations and increased susceptibility to arrhythmias. Recently, we demonstrated that loss of Scn1b in mice leads to compromised mitochondria energetics and reactive oxygen species (ROS) production. In this study, we examined the link between increased ROS and arrhythmia susceptibility in Scn1b-/- mice. In addition, ROS-scavenging capacity can be overwhelmed during prolonged oxidative stress, increasing arrhythmia susceptibility. Therefore, we isolated whole hearts and cardiomyocytes from Scn1b-/- and Scn1b+/+ mice and subjected them to an oxidative challenge with diamide, a glutathione oxidant. Next, we analyzed gene expression and activity of antioxidant enzymes in Scn1b-/- hearts. Cells isolated from Scn1b-/- hearts died faster and displayed higher rates of ROS accumulation preceding cell death compared with those from Scn1b+/+. Furthermore, Scn1b-/- hearts showed higher arrhythmia scores and spent less time free of arrhythmia. Lastly, we found that protein expression and enzymatic activity of glutathione peroxidase is increased in Scn1b-/- hearts compared with wild type. Our results indicate that Scn1b-/- mice have decreased capability to manage ROS during prolonged oxidative stress. ROS accumulation is elevated and appears to overwhelm ROS scavenging through the glutathione system. This imbalance creates the potential for altered cell energetics that may underlie increased susceptibility to arrhythmias or other adverse cardiac outcomes.NEW & NOTEWORTHY Using an oxidative challenge, we demonstrated that isolated cells from Scn1b-/- mice are more susceptible to cell death and surges in reactive oxygen species accumulation. At the whole organ level, they were also more susceptible to the formation of cardiac arrhythmias. This may in part be due to changes to the glutathione antioxidant system.

Intracranial aneurysms in sickle cell disease are associated with hemodynamic stress and anemia.

ABSTRACT: Although hemodynamic stress plays a key role in aneurysm formation outside of sickle cell disease (SCD), its role is understudied in patients with SCD. We hypothesized that tissue-based markers of hemodynamic stress are associated with aneurysm presence in a prospective SCD cohort. Children and adults with SCD, with and without aneurysms, underwent longitudinal brain magnetic resonance imaging/magnetic resonance angiography (MRA) to assess cerebral blood flow (CBF) and oxygen extraction fraction (OEF). Baseline characteristics were recorded. In the subgroup of adults, stepwise mixed-effect logistic regression examined clinical variables, CBF, and OEF as predictors of aneurysm presence. Cumulative rates of new aneurysm formation were estimated using Kaplan-Meier analyses. Forty-three aneurysms were found in 27 of 155 patients (17%). Most aneurysms were ≤3 mm and in the intracranial internal carotid artery. On univariate analysis, older age (P = .07), lower hemoglobin (P = .002), higher CBF (P = .03), and higher OEF (P = .02) were associated with aneurysm presence. On multivariable analysis, age and CBF remained independently associated with aneurysm presence. Seventy-six patients (49% of enrollment) received follow-up MRAs (median, 3.5 years). No aneurysm grew or ruptured, however, 7 new aneurysms developed in 6 patients. The 3-year cumulative rate of aneurysm formation was 3.5%. In 155 patients with SCD, 17% had intracranial aneurysms. Three-year aneurysm formation rate was 3.5%, although limited by small longitudinal sample size and short follow-up duration. Aneurysm presence was associated with elevated CBF in adults, as a tissue-based marker of cerebral hemodynamic stress. Future studies may examine the predictive role of CBF in aneurysm development in SCD.

Return on Investment of Three-Year Accelerated Programs for Students, Medical Schools, Departments, and Community.

Building on the initial accelerated pathway programs in the 1970s to increase workforce, nearly 30 schools have launched accelerated 3-year pathways (A3YP) during the past decade. The authors based on their educational roles, experiences, and scholarship with A3YP provide this perspective of the argument for A3YP and potential disadvantages for each group-students, schools, residencies, departments, and community. When schools consider innovations, they might consider A3YPs for multiple reasons; this perspective helps provide justification for the program and broadly considers return on investment (ROI). The ROI for students includes decreased debt, reduced costs and stress associated with the fourth-year residency applications, and a directed pathway with facilitated transition into a residency program with accompanying professional identity development. Disadvantages for students include early specialty commitment, risk of deceleration, and condensed curriculum. The ROI for schools includes recruiting and retaining students, who will then transition more easily into residency and stimulating innovation. Residency programs gain residents with known skills, who have been a part of the department for 3 years. In addition, fewer residency slots for interviewing leads to saving recruitment administrative costs and time. Finally, many programs are intended to increase the workforce, since students who come to the region for medical school and transition directly into residency are likely to stay in the region. Disadvantages include increased curricular complexity for the medical school, increased administrative support, and advising resources. Finally, several of the accelerated programs attract matriculants from diverse backgrounds contributing to the diversity of the medical school, residency program, and community workforce.

Patient, Parental, and Health Professional Perspectives on Growth in Children With CKD.

RATIONALE & OBJECTIVE: Growth failure is a common problem among children with chronic kidney disease (CKD). Reduced height is associated with psychosocial burden, social stigma, and impaired quality of life. This study aimed to describe the aspects of growth impairment that are most impactful from the perspectives of children with CKD, their parents, and health professionals. STUDY DESIGN: Qualitative study. SETTINGS & PARTICIPANTS: 120 children with CKD (aged 8-21 years), 250 parents, and 445 health professionals from 53 countries participated in 16 focus groups, two consensus workshops, and a Delphi survey. ANALYTICAL APPROACH: A thematic analysis of all qualitative data concerning growth from the Standardized Outcomes in Nephrology - Children and Adolescents (SONG-Kids) initiative. RESULTS: We identified five themes: diminishing psychological wellbeing (compared to and judged by peers, tired of explaining to others, damaging self-esteem), constrained life participation and enjoyment (deprived of normal school experiences, excluded from sports or competing at a disadvantage, impaired quality of life in adulthood); grappling with impacts of symptoms and treatment (difficulty understanding short stature and accessing help, lack of appetite, uncertainty regarding bone pains, medication side effects, burden of growth hormone treatment); facilitating timely interventions and optimizing outcomes (early indicator of disease, assessing management, maximizing transplant outcomes, minimizing morbidity); and keeping growth and health priorities in perspective (quality of life and survival of utmost priority, achieved adequate height). LIMITATIONS: Only English-speaking participants were included. CONCLUSIONS: Impaired growth may diminish psychological wellbeing, self-esteem, and participation in daily activities for children with CKD. Balancing different treatments that can affect growth complicates decision-making. These findings may inform the psychosocial support needed by children with CKD and their caregivers to address concerns about growth.

Stigma and Barriers to Care, Online Mental Health Tools, and Suicidality in California's Veterans.

INTRODUCTION: Veterans have a higher national suicide rate than non-veterans (31.6 versus 18.0 per 100,000). Psychotherapy and other treatments are effective at reducing suicidality, yet stigma and barriers to care reduce willingness to seek help. For veterans who do seek help, they are often undertreated leaving them still in need of help. Online mental health tools (OMHTs) provide another option for obtaining help; however, there is limited research regarding the relationship between stigma and barriers to care, OMHT use, and suicidality. We hypothesized that stigma and barriers are related to higher likelihood of OMHT use and OMHT use is related to lower likelihood of suicidality. MATERIALS AND METHODS: The California Health Interview Survey is a population-based state health survey that collects data over a 2-year cycle via web and phone interviews. The sample was 4,435 veterans (91% male, 75% White, and average age 67 years old). The study was institutional review board exempt because data storage and analyses were done at the Data Access Center at University of California-Los Angeles. RESULTS: Logistic regressions showed endorsing stigma and barriers indicated a higher likelihood of using OMHTs. Additionally, use of OMHTs was unexpectedly associated with greater suicidality. CONCLUSIONS: Findings reinforce the need for research aimed at identifying ways to reduce stigma and barriers toward seeking help. Online mental health tools are a viable option for individuals experiencing stigma and barriers and for individuals who previously experienced suicidality.

RAndomized Cluster Evaluation of Cardiac ARrest Systems (RACE-CARS) trial: Study rationale and design.

Out-of-hospital cardiac arrest (OHCA) occurs in nearly 350,000 people each year in the United States (US). Despite advances in pre and in-hospital care, OHCA survival remains low and is highly variable across systems and regions. The critical barrier to improving cardiac arrest outcomes is not a lack of knowledge about effective interventions, but rather the widespread lack of systems of care to deliver interventions known to be successful. The RAndomized Cluster Evaluation of Cardiac ARrest Systems (RACE-CARS) trial is a 7-year pragmatic, cluster-randomized trial of 62 counties (57 clusters) in North Carolina using an established registry and is testing whether implementation of a customized set of strategically targeted community-based interventions improves survival to hospital discharge with good neurologic function in OHCA relative to control/standard care. The multifaceted intervention comprises rapid cardiac arrest recognition and systematic bystander CPR instructions by 9-1-1 telecommunicators, comprehensive community CPR training and enhanced early automated external defibrillator (AED) use prior to emergency medical systems (EMS) arrival. Approximately 20,000 patients are expected to be enrolled in the RACE CARS Trial over 4 years of the assessment period. The primary endpoint is survival to hospital discharge with good neurologic outcome defined as a cerebral performance category (CPC) of 1 or 2. Secondary outcomes include the rate of bystander CPR, defibrillation prior to arrival of EMS, and quality of life. We aim to identify successful community- and systems-based strategies to improve outcomes of OHCA using a cluster randomized-controlled trial design that aims to provide a high level of evidence for future application.