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Chimeric antigen receptor (CAR)–modified T-cell therapies targeting CD19 represent a new treatment option for patients with relapsed/refractory (R/R) B-cell malignancies. However, CAR T-cell therapy fails to elicit durable responses in a significant fraction of patients. Limited in vivo proliferation and survival of infused CAR T cells are key causes of failure. In a phase 1/2 clinical trial of CD19 CAR T cells for B-cell malignancies (#NCT01865617), low serum interleukin 15 (IL-15) concentration after CAR T-cell infusion was associated with inferior CAR T-cell kinetics. IL-15 supports T-cell proliferation and survival, and therefore, supplementation with IL-15 may enhance CAR T-cell therapy. However, the clinical use of native IL-15 is challenging because of its unfavorable pharmacokinetic (PK) and toxicity. NKTR-255 is a polymer-conjugated IL-15 that engages the entire IL-15 receptor complex (IL-15Rα/IL-2Rβγ) and exhibits reduced clearance, providing sustained pharmacodynamic (PD) responses. We investigated the PK and immune cell PDs in nonhuman primates treated with NKTR-255 and found that NKTR-255 enhanced the in vivo proliferation of T cells and natural killer cells. In vitro, NKTR-255 induced dose-dependent proliferation and accumulation of human CD19 CAR T cells, especially at low target cell abundance. In vivo studies in lymphoma-bearing immunodeficient mice demonstrated enhanced antitumor efficacy of human CD19 CAR T cells. In contrast to mice treated with CAR T cells alone, those that received CAR T cells and NKTR-255 had markedly higher CAR T-cell counts in the blood and marrow that were sustained after tumor clearance, without evidence of persistent proliferation or ongoing activation/exhaustion as assessed by Ki-67 and inhibitory receptor coexpression. These data support an ongoing phase 1 clinical trial of combined therapy with CD19 CAR T cells and NKTR-255 for R/R B-cell malignancies.

Neutrophil extracellular trap (NET) release plays a key role in many chronic disease settings, including atherosclerosis. They are critical to innate immune defence, but also contribute to disease by promoting thrombosis and inflammation. Macrophages are known to release extracellular traps or “METs”, but their composition and role in pathological processes are less well defined. In this study, we examined MET release from human THP-1 macrophages exposed to model inflammatory and pathogenic stimuli, including tumour necrosis factor α (TNFα), hypochlorous acid (HOCl) and nigericin. In each case, there was release of DNA from the macrophages, as visualized by fluorescence microscopy with the cell impermeable DNA binding dye SYTOX green, consistent with MET formation. Proteomic analysis on METs released from macrophages exposed to TNFα and nigericin reveals that they are composed of linker and core histones, together with a range of cytosolic and mitochondrial proteins. These include proteins involved in DNA binding, stress responses, cytoskeletal organisation, metabolism, inflammation, anti-microbial activity, and calcium binding. Quinone oxidoreductase in particular, was highly abundant in all METs but has not been reported previously in NETs. Moreover, there was an absence of proteases in METs in contrast to NETs. Some of the MET histones, contained post-translational modifications, including acetylation and methylation of Lys but not citrullination of Arg. These data provide new insight into the potential implications of MET formation in vivo and their contributions to immune defence and pathology.

Mental illness stigma is a barrier to engagement in mental health services. This study assesses our hypothesis that specific ethnic identity dimensions influences mental health behavior including stigma.We performed an online cross sectional observational study among Black adults (n = 248, ages 18-65). We examined the relationship between an individual's approach to their racial identity in the community and stigma behavior towards mental health; generalized linear models were performed. We assessed demographic characteristics as moderators of the primary association.Black adults with higher centrality reported lower past stigma behavior (RR=1.57, CI: 1.11-2.21,Centrality was associated with mental health stigma behavior. By understanding the intersecting characteristics that may increase the likelihood for mental illness stigma, we will be better able to reduce mental illness stigma and optimize engagement in mental health services.

In the province of Ontario, Canada, it is estimated that 80% of archaeological sites are Indigenous, yet there are very few Indigenous archaeologists involved in management and decision-making about Indigenous heritage. Systemic barriers, particularly around licensing and regulations for curatorial facilities, continue to prevent Indigenous people from directly managing and protecting their own cultural heritage. Recognizing that Indigenous communities in northeastern Ontario have had little exposure or opportunity to learn about archaeology, for several years we have been involved in educational programs to train Indigenous youth, staff in First Nations government offices, and others. We illustrate with two examples: the monitor training program undertaken in northeastern Ontario and a project to catalog artifacts from the legacy collection from the La Cloche site.

CrAssphage and related viruses of the order Crassvirales (hereafter referred to as crassviruses) were originally discovered by cross-assembly of metagenomic sequences. They are the most abundant viruses in the human gut, are found in the majority of individual gut viromes, and account for up to 95% of the viral sequences in some individuals1–4. Crassviruses are likely to have major roles in shaping the composition and functionality of the human microbiome, but the structures and roles of most of the virally encoded proteins are unknown, with only generic predictions resulting from bioinformatic analyses4,5. Here we present a cryo-electron microscopy reconstruction of Bacteroides intestinalis virus ΦcrAss0016, providing the structural basis for the functional assignment of most of its virion proteins. The muzzle protein forms an assembly about 1 MDa in size at the end of the tail and exhibits a previously unknown fold that we designate the ‘crass fold’, that is likely to serve as a gatekeeper that controls the ejection of cargos. In addition to packing the approximately 103 kb of virus DNA, the ΦcrAss001 virion has extensive storage space for virally encoded cargo proteins in the capsid and, unusually, within the tail. One of the cargo proteins is present in both the capsid and the tail, suggesting a general mechanism for protein ejection, which involves partial unfolding of proteins during their extrusion through the tail. These findings provide a structural basis for understanding the mechanisms of assembly and infection of these highly abundant crassviruses.

A critical issue in mine waste management is the management and mitigation of acid rock drainage and metal leaching particularly from waste rock. This requires good understanding and characterization of materials most likely to generate acid pH waters and generate leachable metals and metalloids, as well as the presence of minerals most likely to neutralize acidic pH such as carbonates and certain silicates. For low-sulfidation epithermal deposits in silica-rich volcanic rocks, neutralization minerals are rare. Consequently, determination of acid generation is almost entirely a function of sulfide content of the rocks. By understanding the distribution of sulfide minerals within such a deposit, an initial mine waste management plan can be developed. The Rochester mine currently exploits a low-sulfidation epithermal gold-silver deposit and is located on the southern flank of the Humboldt Range in west-central Nevada. The Rochester mine is a conventional open-pit mining operation with cyanide heap leaching and Merrill Crowe processing. Silver and lesser gold mineralization occur associated with sulfide minerals hosted in Permian-Triassic Koipato Group rhyolitic units with extensive quartz-sericite-pyrite alteration. Gangue mineralogy of the deposit is dominated by slow or unreactive silicate minerals with little or no carbonate. Silver-bearing mineralization was controlled by primary hypogene sulfides and secondary supergene oxidation processes, and most of the mineralized material mined to date has been fully or partially oxidized. Oxidation mobilizes and depletes all forms of sulfur from primary sulfide minerals. Due to the low sulfate content and low content of neutralizing gangue phases, the waste rock classification can be based on total sulfur content. Based on the analysis of pulp samples collected from within the mine area, a sulfur block model was developed and compared to the results of standard acid-base accounting and net acid generation tests. This comparison demonstrates the benefit of the block model in identifying zones of potentially acid-generating material in the mined waste. Application of this information to mine planning has allowed more efficient management of acid-generating mine waste by allowing the operator to develop areas of the waste rock facility to receive the waste in a timely fashion. Such an approach is based on good mine geologic and mineralogical characterization and the use of a representative and comprehensive geologic block model.

PURPOSE Novel biomarkers are needed to differentiate outcomes in intermediate-risk rhabdomyosarcoma (IR RMS). We sought to evaluate strategies for identifying circulating tumor DNA (ctDNA) in IR RMS and to determine whether ctDNA detection before therapy is associated with outcome. PATIENTS AND METHODS Pretreatment serum and tumor samples were available from 124 patients with newly diagnosed IR RMS from the Children's Oncology Group biorepository, including 75 patients with fusion-negative rhabdomyosarcoma (FN-RMS) and 49 with fusion-positive rhabdomyosarcoma (FP-RMS) disease. We used ultralow passage whole-genome sequencing to detect copy number alterations and a new custom sequencing assay, Rhabdo-Seq, to detect rearrangements and single-nucleotide variants. RESULTS We found that ultralow passage whole-genome sequencing was a method applicable to ctDNA detection in all patients with FN-RMS and that ctDNA was detectable in 13 of 75 serum samples (17%). However, the use of Rhabdo-Seq in FN-RMS samples also identified single-nucleotide variants, such as MYOD1 L122R, previously associated with prognosis. Identification of pathognomonic translocations between PAX3 or PAX7 and FOXO1 by Rhabdo-Seq was the best method for measuring ctDNA in FP-RMS and detected ctDNA in 27 of 49 cases (55%). Patients with FN-RMS with detectable ctDNA at diagnosis had significantly worse outcomes than patients without detectable ctDNA (event-free survival, 33.3% v 68.9%; P = .0028; overall survival, 33.3% v 83.2%; P < .0001) as did patients with FP-RMS (event-free survival, 37% v 70%; P = .045; overall survival, 39.2% v 75%; P = .023). In multivariable analysis, ctDNA was independently associated with worse prognosis in FN-RMS but not in the smaller FP-RMS cohort. CONCLUSION Our study demonstrates that baseline ctDNA detection is feasible and is prognostic in IR RMS.

Objective To quantify the financial and clinical implications of pharmacists completing Medicare Annual Wellness Visits (AWV). Design Retrospective, observational, population-based health economic analysis utilizing electronic medical record data. Setting A single family-practice clinic within a Missouri-based not-for-profit community health system. Patients/Participants Medicare patients in their second year of coverage who had not been seen for an AWV within the previous 12 months. Intervention Patients attended a subsequent AWV conducted by a licensed ambulatory care pharmacist between July 1 and December 31, 2021. The pharmacist completed all necessary visit requirements and helped facilitate completion of preventive screenings. Main Outcome Measures Primary outcomes measured were financial implications of pharmacists completing AWVs. The primary objective was to quantify the direct and indirect financial benefits for a health system incorporating pharmacist-led AWVs. Direct revenue was calculated based on the number of visits completed multiplied by the reimbursement rate paid to the health care system by the Centers for Medicare & Medicaid Services (CMS). Indirect revenue was estimated based on the number and type of interventions completed by the patient. The secondary outcome measured was the clinical impact of pharmacist-led AWVs through quantifying preventive care service needs and completion of these services. Alongside these measures, the number of medication discrepancies and drug interactions were also measured and cost avoidance estimated. Annual cost in salary of a pharmacist was considered, but additional cost of overhead was beyond the scope of the study. Results Thirty-nine AWVs were completed. Two hundred six recommended screenings, labs, and immunizations were identified, of which 24% were completed. The pharmacists were also able to identify 205 medication issues. These visits generated $4,593.42 in direct revenue. Recommended preventive care generated $2,161.71 in indirect revenue. Conclusion Pharmacist-led AWVs are financially beneficial to the health care system and increases the number of completed AWVs while avoiding demand for additional physician time.

This study examined the experiences of immigrant parents seeking asylum in the United States after recently fleeing the Northern Triangle region of Central America, attending both to the traumatic events and the strengths that mitigate this experience. The study employs a convergent parallel mixed-methods design to describe the experiences of 51 parents after being processed by U.S. immigration officials for seeking asylum. Quantitative analyses measured trauma exposure and psychological distress and posttraumatic stress symptoms, as well as religiosity, dispositional optimism, and perceived quality of life. Qualitative analyses examined migratory experiences more deeply. Quantitative findings suggest that participants displayed high levels of resilience as evidenced by the low endorsement of mental health symptoms associated with trauma despite high violence and trauma exposure. Qualitative analyses revealed two themes underscoring strengths that mitigated the compounded trauma experienced by participants, including: (a) protective factors, and (b) plans for the future. In addition, several subthemes emerged within each of these broader themes. Qualitative findings highlight that asylum seekers come to the United States for the sake of their families, especially their children. Participants shared their desire to ensure the safety of their family and children and their hope of being better able to take care of them. These desires and hopes common among participants in our study not only motivated them to leave their home countries but also helped them to press forward in their journeys. Study results show that religiosity, hope, and optimism also played important roles in sustaining these immigrant parents during their journeys.

This study aimed to comprehensively assess the pattern of cortical maturation by magnetic resonance imaging (MRI) in fetuses with unaffected and mildly affected CMV infection determined by ultrasound (US) and establish possible differences compared to healthy controls.Twenty-four CMV-infected fetuses (7 US unaffected, 17 US mildly affected) and 24 healthy controls between 27 and 36 weeks of gestation undergoing fetal MRI were included. Fetuses were considered unaffected or mildly affected according to prenatal US/MRI neuroimaging findings. We compared fetal sulci depth, Sylvian fissure depth, Sylvian fissure, and angle and cortical maturation grading of specific sulci and areas among study groups. Regression, parametric trend and intraclass correlation analysis were performed.Compared to controls, in the CMV-infected fetuses showed a significantly larger median (interquartile range: IQR) width of the lateral ventricles [right (mm): 3.9 (2.6-5.3) vs. 7.8 (5.9-9.9); left: 4.2 (3.2-5.3) vs. 7.5 (6.0-10.9)], significantly decreased parieto-occipital sulcus [right (mm): 15.9 (14.7-17.3) vs. 12.6 (11.3-13.5); left: 16.0 (13.3-17.5) vs. 12.3 (10.6-13.5)] and calcarine sulcus depth [right (mm): 17.5 (16.1-18.7) vs. 15.4 (14.4-16.3); left: 16.7 (15.6-18.9) vs. 14.6 (14.1-15.6)], p0.001; and a significantly larger upper and lower Sylvian fissure angle [upper, right (°): 42.8 (35.8-45.8) vs. 48.9 (38.4-64.7); left: 40.9 (34.2-45.8) vs. 48.2 (41.9-60.7)], [lower, right: 41.6 (34.4-49.2) vs. 48.9 (40.6-60.9); left: 42.2 (38.8-46.9) vs. 48.9 (39.5-57.5)]; p0.05. In addition, the infected fetuses had a significantly lower cortical-grading in the temporal and parietal areas, and the parieto-occipital and calcarine sulcus compared to healthy fetuses (p0.05). These differences persisted when adjusting for gestational age, ipsilateral atrium width, fetal gender and considering being small for gestational age as a confounding/interacting factor.Unaffected and mildly affected CMV-infected fetuses with mild sonographic involvement showed underdeveloped cortical maturation compared to healthy controls. These results suggest that congenital CMV-infection, even in non-severely affected fetuses, which are typically considered of good prognosis, could be related to altered brain cortical structure. Further research is warranted to better elucidate its correlation with neurodevelopmental outcomes. This article is protected by copyright. All rights reserved.

Objectives. To examine the impact of school shootings on indicators of adolescent school safety in the United States. Methods. We linked 2009–2019 Youth Risk Behavior Survey data on 211 236 adolescents aged 14 to 18 years from 24 school districts with data on high school shootings from the Center for Homeland Defense and Security. We conducted 2-way fixed-effects logistic regression models to assess the impact of shootings on self-report of 3 indicators of school safety: avoiding school because of feeling unsafe, carrying a weapon at school, and being threatened or injured with a weapon at school. Results. High school shootings were associated with adolescents having 20% greater odds of avoiding school because of feeling unsafe (adjusted odd ratio [AOR] = 1.20; 95% confidence interval [CI] = 1.11, 1.29) than those who had not. Findings were slightly attenuated in sensitivity analyses that tested exposure to shootings at any school in the district or state. High school shootings were associated with a statistically nonsignificant (P = .08) elevated risk of carrying a weapon at school (AOR = 1.11; 95% CI = 0.99, 1.25). Conclusions. The negative ramifications of school shootings extend far beyond the event itself to adolescents’ concerns about school safety. (Am J Public Health. 2023;113(4):438–441. https://doi.org/10.2105/AJPH.2022.307206 )

The detection of gene fusions by RNA-based next-generation sequencing (NGS) is an emerging method in clinical genetic laboratories for oncology biomarker testing to direct targeted therapy selections. A recent Canadian study (CANTRK study) comparing the detection of NTRK gene fusions on different NGS assays to determine subjects’ eligibility for tyrosine kinase TRK inhibitor therapy identified the need for recommendations for best practices for laboratory testing to optimize RNA-based NGS gene fusion detection. To develop consensus recommendations, representatives from 17 Canadian genetic laboratories participated in working group discussions and the completion of survey questions about RNA-based NGS. Consensus recommendations are presented for pre-analytic, analytic and reporting aspects of gene fusion detection by RNA-based NGS.

Purpose The purpose of the study was to examine the relationship between satisfaction of Medicare coverage for out-of-pocket costs and problems paying medical bills among Medicare beneficiaries with type 2 diabetes. Methods The 2019 Medicare Current Beneficiary Survey Public Use File, a nationally representative sample of Medicare beneficiaries aged ≥65 years with type 2 diabetes, was analyzed (n = 2178). A survey-weighted multivariable logit regression model was conducted to examine the association between satisfaction of Medicare coverage for out-of-pocket costs and problems paying medical bills, adjusted for sociodemographics and comorbidities. Results Among study beneficiaries, 12.6% reported problems paying medical bills. Among those with and without problems paying medical bills, 59.5% and 12.8%, respectively, were dissatisfied with out-of-pocket costs. In the multivariable analysis, beneficiaries who were dissatisfied with out-of-pocket costs were more likely to report problems paying medical bills than those who were satisfied. Younger beneficiaries, beneficiaries with lower incomes, those with functional limitations, and those with multiple comorbidities were more likely to report problems paying medical bills. Conclusions Despite having health care coverage, more than one-tenth of Medicare beneficiaries with type 2 diabetes reported problems paying medical bills, which raises concerns about delaying or forgoing needed medical care due to unaffordability. Screenings and targeted interventions that identify and reduce financial hardships associated with out-of-pocket costs should be prioritized.