Aconitate decarboxylase 1 mediates the acute airway inflammatory response to environmental exposures.

Background: Environmental lipopolysaccharide (LPS) and microbial component-enriched organic dusts cause significant lung disease. These environmental exposures induce the recruitment and activation of distinct lung monocyte/macrophage subpopulations involved in disease pathogenesis. Aconitate decarboxylase 1 (Acod1) was one of the most upregulated genes following LPS (vs. saline) exposure of murine whole lungs with transcriptomic profiling of sorted lung monocyte/macrophage subpopulations also highlighting its significance. Given monocyte/macrophage activation can be tightly linked to metabolism, the objective of these studies was to determine the role of the immunometabolic regulator ACOD1 in environmental exposure-induced lung inflammation. Methods: Wild-type (WT) mice were intratracheally (i.t.) instilled with 10 µg of LPS or saline. Whole lungs were profiled using bulk RNA sequencing or sorted to isolate monocyte/macrophage subpopulations. Sorted subpopulations were then characterized transcriptomically using a NanoString innate immunity multiplex array 48 h post-exposure. Next, WT and Acod1-/- mice were instilled with LPS, 25% organic dust extract (ODE), or saline, whereupon serum, bronchoalveolar lavage fluid (BALF), and lung tissues were collected. BALF metabolites of the tricarboxylic acid (TCA) cycle were quantified by mass spectrometry. Cytokines/chemokines and tissue remodeling mediators were quantitated by ELISA. Lung immune cells were characterized by flow cytometry. Invasive lung function testing was performed 3 h post-LPS with WT and Acod1-/- mice. Results: Acod1-/- mice treated with LPS demonstrated decreased BALF levels of itaconate, TCA cycle reprogramming, decreased BALF neutrophils, increased lung CD4+ T cells, decreased BALF and lung levels of TNF-α, and decreased BALF CXCL1 compared to WT animals. In comparison, Acod1-/- mice treated with ODE demonstrated decreased serum pentraxin-2, BALF levels of itaconate, lung total cell, neutrophil, monocyte, and B-cell infiltrates with decreased BALF levels of TNF-α and IL-6 and decreased lung CXCL1 vs. WT animals. Mediators of tissue remodeling (TIMP1, MMP-8, MMP-9) were also decreased in the LPS-exposed Acod1-/- mice, with MMP-9 also reduced in ODE-exposed Acod1-/- mice. Lung function assessments demonstrated a blunted response to LPS-induced airway hyperresponsiveness in Acod1-/- animals. Conclusion: Acod1 is robustly upregulated in the lungs following LPS exposure and encodes a key immunometabolic regulator. ACOD1 mediates the proinflammatory response to acute inhaled environmental LPS and organic dust exposure-induced lung inflammation.

Establishing the minimal clinically important difference of the Brief Fatigue Inventory for brain or CNS cancer patients undergoing radiotherapy.

Background: Minimal clinically important differences (MCIDs) quantify the clinical relevance of quality of life results at the individual patient and group level. The aim of this study was to estimate the MCID for the Brief Fatigue Inventory (BFI) and the Worst and Usual Fatigue items in patients with brain or CNS cancer undergoing curative radiotherapy. Methods: Data from a multi-site prospective registry was used. The MCID was calculated using distribution-based and anchor-based approaches. For the anchor-based approach, the fatigue item from the PROMIS-10 served as the anchor to determine if a patient improved, deteriorated, or had no change from baseline to end of treatment (EOT). We compared the unadjusted means on the BFI for the 3 groups to calculate the MCID. For the distribution-based approaches, we calculated the MCID as 0.5 SD of the scores and as 1.96 times the standard error of measurement. Results: Three-hundred and fifty nine patients with brain or CNS tumors undergoing curative radiotherapy filled out the 9-item BFI at baseline and EOT. The MCID for the BFI was 1.33 (ranging from 0.99 to 1.70 across the approaches), 1.51 (ranging from 1.16 to 2.02) and 1.76 (ranging from 1.38 to 2.14) for the usual and worst fatigue items, respectively. Conclusions: This study provides the MCID ranges for the BFI and Worst and Usual fatigue items, which will allow clinically meaningful conclusions to be drawn from BFI scores. These results can be used to select optimal treatments for patients with brain or CNS cancer or to interpret BFI scores from clinical trials.

Physiological and perceptual responses to sprint interval exercise using arm versus leg cycling ergometry.

Increases in power output and maximal oxygen consumption ( V ˙ O 2 max) occur in response to sprint interval exercise (SIE), but common use of "all-out" intensities presents a barrier for many adults. Furthermore, lower-body SIE is not feasible for all adults. We compared physiological and perceptual responses to supramaximal, but "non-all-out" SIE between leg and arm cycling exercise. Twenty-four active adults (mean â€‹± â€‹SD age: [25 â€‹± â€‹7] y; cycling V ˙ O 2 max: [39 â€‹± â€‹7] mL·kg-1·min-1) performed incremental exercise using leg (LCE) and arm cycle ergometry (ACE) to determine V ˙ O 2 max and maximal work capacity (Wmax). Subsequently, they performed four 20 â€‹s bouts of SIE at 130% Wmax on the LCE or ACE at cadence â€‹= â€‹120-130 â€‹rev/min, with 2 â€‹min recovery between intervals. Gas exchange data, heart rate (HR), blood lactate concentration (BLa), rating of perceived exertion (RPE), and affective valence were acquired. Data showed significantly lower (p â€‹< â€‹0.001) absolute mean ([1.24 â€‹± â€‹0.31] L·min-1 vs. [1.59 â€‹± â€‹0.34] L·min-1; d â€‹= â€‹1.08) and peak V ˙ O 2 ([1.79 â€‹± â€‹0.48] L·min-1 vs. [2.10 â€‹± â€‹0.44] L·min-1; d â€‹= â€‹0.70) with ACE versus LCE. However, ACE elicited significantly higher (p â€‹< â€‹0.001) relative mean ([62% â€‹± â€‹9%] V ˙ O 2 max vs. [57% â€‹± â€‹7%] V ˙ O 2 max, d â€‹= â€‹0.63) and peak V ˙ O 2 ([88% â€‹± â€‹10%] V ˙ O 2 max vs. [75% â€‹± â€‹10%] V ˙ O 2 max, d â€‹= â€‹1.33). Post-exercise BLa was significantly higher ([7.0 â€‹± â€‹1.7] mM vs. [5.7 â€‹± â€‹1.5] mM, p â€‹= â€‹0.024, d â€‹= â€‹0.83) for LCE versus ACE. There was no significant effect of modality on RPE or affective valence (p â€‹> â€‹0.42), and lowest affective valence recorded (2.0 â€‹± â€‹1.8) was considered "good to fairly good". Data show that non "all-out" ACE elicits lower absolute but higher relative HR and V ˙ O 2 compared to LCE. Less aversive perceptual responses could make this non-all-out modality feasible for inactive adults.

Increases in the incremental exercise mean response time across the steady state domain: Implications for exercise testing & prescription.

We hypothesized that slowed oxygen uptake ( V ˙ O 2 ) kinetics for exercise transitions to higher power outputs (PO) within the steady state (SS) domain would increase the mean response time (MRT) with increasing exercise intensity during incremental exercise. Fourteen highly trained cyclists (mean ±â€¯standard deviation [SD]; age (39 ±â€¯6) years [yr]; and V ˙ O 2 peak = (61 ±â€¯9) mL/kg/min performed a maximal, ramp incremental cycling test and on separate days, four 6-min bouts of cycling at 30%, 45%, 65% & 75% of their incremental peak PO (Wpeak). SS trial data were used to calculate the MRT and verified by mono-exponential and linear curve fitting. When the ramp protocol attained the value from SS, the PO, in Watts (W), was converted to time (min) based on the ramp function W to quantify the incremental MRT (iMRT). Slope analyses for the V ˙ O 2 responses of the SS versus incremental exercise data below the gas exchange threshold (GET) revealed a significant difference (p = 0.003; [0.437 ±â€¯0.08] vs. [0.382 ±â€¯0.05] L⋅min-1). There was a significant difference between the 45% Wpeak steady state V ˙ O 2 (ss V ˙ O 2 ) ([3.08 ±â€¯0.30] L⋅min-1, respectively), and 30% Wpeak ss V ˙ O 2 (2.26 ±â€¯0.24) (p < 0.0001; [3.61 ±â€¯0.80] vs. [2.20 ±â€¯0.39] L⋅min-1) and between the iMRT for 45% and 30% Wpeak ss V ˙ O 2 values ([50.58 ±â€¯36.85] s vs. [32.20 ±â€¯43.28] s). These data indicate there is no single iMRT, which is consistent with slowed V ˙ O 2 kinetics and an increasing V ˙ O 2 deficit for higher exercise intensities within the SS domain.

Palbociclib in Solid Tumor Patients With Genomic Alterations in the cyclinD-cdk4/6-INK4a-Rb Pathway: Results From National Cancer Institute-Children's Oncology Group Pediatric Molecular Analysis for Therapy Choice Trial Arm I (APEC1621I).

PURPOSE: The National Cancer Institute-Children's Oncology Group Pediatric Molecular Analysis for Therapy Choice trial assigned patients age 1-21 years with relapsed or refractory solid tumors, lymphomas, and histiocytic disorders to phase II treatment arms of molecularly targeted therapies on the basis of genetic alterations detected in their tumor. Patients with tumors that harbored prespecified genomic alterations in the cyclinD-CDK4/6-INK4a-Rb pathway with intact Rb expression were assigned and treated with the cdk4/6 inhibitor palbociclib. METHODS: Patients received palbociclib orally once daily for 21 days of 28-day cycles until disease progression, intolerable toxicity, or up to 2 years. The primary end point was objective response rate; secondary end points included safety/tolerability and progression-free survival. RESULTS: Twenty-three patients (median age, 15 years; range, 8-21) were enrolled; 20 received protocol therapy and were evaluable for toxicity and response. Of the evaluable patients, the most common diagnoses were osteosarcoma (n = 9) and rhabdomyosarcoma (n = 6). A single actionable gene amplification was found in 19 tumors (CDK4, n = 11, CDK6, n = 2, CCND3, n = 6), with one tumor harboring two amplifications (CDK4 and CCND2). Hematologic toxicities were the most common treatment-related events. No objective responses were seen. Two patients with tumors harboring CDK4 amplifications (neuroblastoma and sarcoma) had best response of stable disease for six and three cycles. Six-month progression was 10% (95% CI, 1.7 to 27.2). CONCLUSION: The CDK4/6 inhibitor palbociclib at 75 mg/m2 orally daily was tolerable in this heavily pretreated cohort. No objective responses were observed in this histology-agnostic biomarker-selected population with treatment-refractory solid tumors, demonstrating that pathway alteration alone is insufficient in pediatric cancers to generate a response to palbociclib monotherapy.

Lower limb coordination patterns following anterior cruciate ligament reconstruction: A longitudinal study.

BACKGROUND: Changes in lower limb joint coordination have been shown to increase localized stress on knee joint soft tissue-a known precursor of osteoarthritis. While 50 % of individuals who undergo anterior cruciate ligament reconstruction (ACLR) develop radiographic osteoarthritis, it is unclear how underlying joint coordination during gait changes post-ACLR. The purpose of this study was twofold: to determine differences in lower limb coordination patterns during gait in ACLR individuals 2, 4, and 6 months post-ACLR and to compare the coordination profiles of the ACLR participants at each timepoint post-ACLR to uninjured matched controls. METHODS: We conducted a longitudinal assessment to quantify lower limb coordination at 3 timepoints post-ACLR and compared the ACLR coordination profiles to uninjured controls. Thirty-four ACLR (age = 21.43 ±â€¯4.24 years, mean ±â€¯SD; 70.59 % female) and 34 controls (age = 21.42 ±â€¯3.43 years; 70.59 % female) participated. The ACLR group completed 3 overground gait assessments (2,4, and 6 months post-ACLR), and the controls completed 1 assessment, at which lower limb kinematics were collected. Cross-recurrence quantification analysis was used to characterize sagittal and frontal plane ankle-knee, ankle-hip, and knee-hip coordination dynamics. Comprehensive general linear mixed models were constructed to compare between-limb and within-limb coordination outcomes over time post-ACLR and a between-group comparison across timepoints. RESULTS: The ACLR limb demonstrated a more "stuck" sagittal plane knee-hip coordination profile (greater trapping time (TT); p = 0.004) compared bilaterally. Between groups, the ACLR participants exhibited a more predictable ankle-knee coordination pattern (percent determinism (%DET); p < 0.05), stronger coupling between joints (meanline (MNLine)) across all segments (p < 0.05), and greater knee-hip TT (more "stuck"; p < 0.05) compared to the controls at each timepoint in the sagittal plane. Stronger frontal plane knee-hip joint coupling (MNLine) persisted across timepoints within the ACLR group compared to the controls (p < 0.05). CONCLUSION: The results indicate ACLR individuals exhibit a distinct and rigid coordination pattern during gait compared to controls within 6-month post-ACLR, which may have long-term implications for knee-joint health.

Prevalence and Factors Associated with High Impact Chronic Pain in Knee Osteoarthritis: the Johnston County Health Study.

Pain is a hallmark symptom of knee osteoarthritis (KOA) yet intensity and severity vary widely among individuals. There is a knowledge gap in understanding key characteristics of high impact chronic pain (HICP) within the context of KOA. Therefore, our first purpose was to examine the prevalence of HICP in a cohort of individuals with radiographic evidence of KOA, and our second purpose was to assess patient level factors associated with HICP. Data from the Johnston County Health Study (JoCoHS) were used to compare those with and without HICP. Variables included sociodemographic factors, clinical factors, health care use, and psychosocial distress. HICP status was classified with PROMIS Pain Interference (PI) and Physical Function (PF) measures. Results indicated that 15.5% (48/310) of participants were classified as having HICP when the PROMIS-PI cutoff score was used, while 21.2% (66/310) were classified as having HICP with a PROMIS-PF cutoff score. Multivariable analyses indicated that HICP was consistently characterized by increased kinesiophobia and somatization regardless of PROMIS measure used for HICP status. A secondary insight was that HICP was not consistently characterized by sociodemographic and clinical variables, as these findings were dependent on PROMIS measured used. These findings could be used to develop intervention approaches specific to individuals with KOA and to inform future investigations of sociodemographic and clinical factors associated with HICP. PERSPECTIVE: These findings provide additional information on the characterization of HICP for individuals with KOA. There was consistency in psychosocial factors associated with HICP, while sociodemographic and clinical factors varied based on how HICP status was defined.

Protocol for the longitudinal study of neuroinflammation and reactive astrocytes in Lcn2CreERT2 mice.

During brain disease, astrocytes can reprogram into a reactive state that alters many of their functions. Here, we present a protocol for studying neuroinflammation and reactive astrogliosis in mice using lipopolysaccharide (LPS) from E. coli. We describe steps for employing the Lcn2CreERT2 mouse crossed into a fluorescent Cre reporter line to label a subset of reactive astrocytes during and after inflammation. We then detail procedures for the longitudinal study of reactive astrocytes during the induction, progression, and/or resolution of astrogliosis. For complete details on the use and execution of this protocol, please refer to Agnew-Svoboda et al.1.

Colloidal 2D Layered SiC Quantum Dots from a Liquid Precursor: Surface Passivation, Bright Photoluminescence, and Planar Self-Assembly.

We report the bottom-up synthesis of colloidal two-dimensional (2D) layered silicon carbide (SiC) quantum dots with a cubic structure, lateral size of 5-10 nm, ⟨110⟩ exfoliation to few atomic layers, and surface passivation with 1-dodecene. Samples shielded from oxygen and plasma-annealed for purity exhibit narrow blue photoluminescence (PL) with quantum yields (QYs) over 60% in exceptional cases, while unshielded nanocrystals (NCs) exhibit broad blue/green/white PL with 10-15% QY. The latter scenario is attributed to excess surface carbon and oxygen accrued during synthesis and processing, with size separation through ultracentrifugation revealing size-dependent impurity emission. In contrast, the shape of the bright narrow blue PL shows little variation with NC size, while in both scenarios, the maximum QY occurs near four atomic layers. When dried under heat, the disk-like NC suspensions are observed to aggregate into microscale domains, with further self-assembly into planar superlattice domains with common crystalline orientation. The results are compared with photophysical simulations and bring clarity to the broad emission commonly reported for top-down approaches, while inspiring bottom-up schemes directed at improved material quality.

Older adults are relatively more susceptible to impulsive social influence than young adults.

People differ in their levels of impulsivity and patience, and these preferences are heavily influenced by others. Previous research suggests that susceptibility to social influence may vary with age, but the mechanisms and whether people are more influenced by patience or impulsivity remain unknown. Here, using a delegated inter-temporal choice task and Bayesian computational models, we tested susceptibility to social influence in young (aged 18-36, N = 76) and older (aged 60-80, N = 78) adults. Participants completed a temporal discounting task and then learnt the preferences of two other people (one more impulsive and one more patient) before making their choices again. We used the signed Kullback-Leibler divergence to quantify the magnitude and direction of social influence. We found that, compared to young adults, older adults were relatively more susceptible to impulsive social influence. Factor analyses showed that older adults with higher self-reported levels of affective empathy and emotional motivation were particularly susceptible to impulsive influence. Importantly, older and young adults showed similar learning accuracy about others' preferences, and their baseline impulsivity did not differ. Together, these findings suggest highly affectively empathetic and emotionally motivated older adults may be at higher risk for impulsive decisions, due to their susceptibility to social influence.

Gene expression responses of CF airway epithelial cells exposed to elexacaftor/tezacaftor/ivacaftor (ETI) suggest benefits beyond improved CFTR channel function.

The combination of elexacaftor/tezacaftor/ivacaftor (ETI, Trikafta) reverses the primary defect in Cystic Fibrosis (CF) by improving CFTR mediated Cl - and HCO 3 - secretion by airway epithelial cells (AEC), leading to improved lung function and less frequent exacerbations and hospitalizations. However, studies have shown that CFTR modulators like ivacaftor, a component of ETI, has numerous effects on CF cells beyond improved CFTR channel function. Because little is known about the effect of ETI on CF AEC gene expression we exposed primary human AEC to ETI for 48 hours and interrogated the transcriptome by RNA-seq and qPCR. ETI increased defensin gene expression ( DEFB1 ) an observation consistent with reports of decreased bacterial burden in the lungs of people with CF (pwCF). ETI also decreased MMP10 and MMP12 gene expression, suggesting that ETI may reduce proteolytic induced lung destruction in CF. ETI also reduced the expression of the stress response gene heme oxygenase ( HMOX1 ). qPCR analysis confirmed DEFB1, HMOX1, MMP10 and MMP12 gene expression results observed by RNA-seq. Gene pathway analysis revealed that ETI decreased inflammatory signaling, cellular proliferation and MHC Class II antigen presentation. Collectively, these findings suggest that the clinical observation that ETI reduces lung infections in pwCF is related in part to drug induced increases in DEFB1 , and that ETI may reduce lung damage by reducing MMP10 and MMP12 gene expression, which is predicted to reduce matrix metalloprotease activity. Moreover, pathway analysis also identified several genes responsible for the ETI induced reduction in inflammation observed in people with CF. New and Noteworthy: Gene expression responses by CF AEC exposed to ETI suggest that in addition to improving CFTR channel function, ETI is likely to increase resistance to bacterial infection by increasing levels of beta defensin 1 (hBD-1). ETI may also reduce lung damage by suppressing MMP10, and reduce airway inflammation by repressing proinflammatory cytokine secretion by AEC cells.

Clinician Perspectives on Decision Support and AI-based Decision Support in a Pediatric ED.

BACKGROUND: Clinical decision support (CDS) systems offer the potential to improve pediatric care through enhanced test ordering, prescribing, and standardization of care. Its augmentation with artificial intelligence (AI-CDS) may help address current limitations with CDS implementation regarding alarm fatigue and accuracy of recommendations. We sought to evaluate strengths and perceptions of CDS, with a focus on AI-CDS, through semistructured interviews of clinician partners. METHODS: We conducted a qualitative study using semistructured interviews of physicians, nurse practitioners, and nurses at a single quaternary-care pediatric emergency department to evaluate clinician perceptions of CDS and AI-CDS. We used reflexive thematic analysis to identify themes and purposive sampling to complete recruitment with the goal of reaching theoretical sufficiency. RESULTS: We interviewed 20 clinicians. Participants demonstrated a variable understanding of CDS and AI, with some lacking a clear definition. Most recognized the potential benefits of AI-CDS in clinical contexts, such as data summarization and interpretation. Identified themes included the potential of AI-CDS to improve diagnostic accuracy, standardize care, and improve efficiency, while also providing educational benefits to clinicians. Participants raised concerns about the ability of AI-based tools to appreciate nuanced pediatric care, accurately interpret data, and about tensions between AI recommendations and clinician autonomy. CONCLUSIONS: AI-CDS tools have a promising role in pediatric emergency medicine but require careful integration to address clinicians' concerns about autonomy, nuance recognition, and interpretability. A collaborative approach to development and implementation, informed by clinicians' insights and perspectives, will be pivotal for their successful adoption and efficacy in improving patient care.

Safety, reactogenicity, and immunogenicity of different doses of 10-valent Extraintestinal Pathogenic Escherichia Coli (ExPEC10V) bioconjugate vaccine (VAC52416) in healthy Japanese adults aged 60-85 years in a randomized, double-blind, phase 1 study.

AIM: This phase 1 study (NCT04306302) evaluated the safety, reactogenicity, and immunogenicity of ExPEC10V (VAC52416) in healthy Japanese adults. METHOD: The randomized, double-blind, single-center study included 28-day screening, vaccination (Day 1), 30-day safety and immunogenicity follow-up and 181-day serious adverse events (SAEs) follow-up. Participants (60-85 years) were enrolled in dose-ascending approach and randomized to medium- and high-doses of ExPEC10V (n = 8 in each dose group) and placebo (n = 8). Incidence of adverse events: solicited AEs (until Day 15), unsolicited AEs (until Day 30), SAEs (until Day 181) and immunogenicity (electrochemiluminescent-based assay [ECL] and multiplex opsonophagocytic assay [MOPA]) were assessed on Day 15 and Day 30. RESULTS: Total of 24 participants were included (median age, 66.5 years; 50.0 % female). Incidence of solicited AEs was 81.3 % (local) and 18.8 % (systemic) for pooled ExPEC10V group (medium-dose ExPEC10V: 75.0 % [local], 12.5 % [systemic]; high-dose ExPEC10V: 87.5 % [local], 25.0 % [systemic]). One SAE, not vaccine-related, was reported in high-dose ExPEC10V group after Day 30, which was resolved during study. The ECL demonstrated increase in binding antibody titers, which was maintained from Day 15 to Day 30. For all serotypes, the geometric mean fold increases from baseline on Day 15 ranged from 2.51 to 10.60 and 1.97-5.23 for medium- and high-dose groups, respectively. The MOPA demonstrated increase in functional antibody responses for all serotypes (except O8) at Day 15 which was maintained from Day 15 to Day 30. CONCLUSIONS: ExPEC10V medium- and high-doses were well tolerated with an acceptable safety profile without any significant safety issues in healthy Japanese participants.

Concomitant Use of Oral Anticoagulants with Oral Dipeptidyl Peptidase-4 Inhibitors and Serious Bleeding Events.

In a prior screening study, saxagliptin, a dipeptidyl peptidase-4 inhibitor (DPP-4i), was found to have an increased rate of serious bleeding when used concomitantly with several oral anticoagulants (OACs). We aimed to confirm or refute the associations between concomitant use of individual OACs and DPP-4is and serious bleeding in a large US database, using self-controlled case series (SCCS) and case-crossover (CCO) designs. The study population was eligible Medicare beneficiaries co-exposed to a DPP-4i (precipitant) and either an OAC (object drug) or lisinopril (negative control object drug) in 2016-2020. For the SCCS, we used conditional Poisson regression to estimate adjusted rate ratios (RRs) between each co-exposure (vs. not) and serious bleeding and divided the RR by the adjusted RR for the corresponding lisinopril + precipitant pair to obtain ratios of RRs (RRRs). For the CCO, we estimated the adjusted odds ratios (ORs) of exposure to the precipitant in the focal window vs. referent window using multivariable conditional logistic regression and divided the ORs in the object drug-exposed cases over the ORs in negative object drug-exposed cases to obtain the ratios of ORs (RORs). The adjusted RRRs for serious bleeding ranged from 0.32 (0.05-1.91) for apixaban/lisinopril + saxagliptin to 3.49 (1.29-9.48) for warfarin/lisinopril + linagliptin. The adjusted RORs ranged from 0.01 (0.00-0.20) for rivaroxaban/lisinopril + saxagliptin to 2.99 (0.74-12.11) for apixaban/lisinopril + linagliptin. While we could not confirm previously identified signals because of statistical imprecision, several numerically elevated estimates still warrant caution in concomitant use and further examination.

MicroRNA-146a deficiency enhances host protection against murine cytomegalovirus.

Natural killer (NK) cells are innate lymphoid cells that protect a host from viral infections and malignancies. MicroRNA-146a (miR-146a) is an important regulator of immune function that is highly expressed in NK cells and is further upregulated during murine cytomegalovirus (MCMV) infection. Here we utilized mice with a global targeted deletion of miR-146a to understand its impact on the innate immune responses to MCMV infection. MiR-146a-/- mice were protected from lethal MCMV infection, which was intrinsic to the hematopoietic compartment based on bone marrow chimera experiments. NK cell depletion abrogated this protection, implicating NK cells as critical for the miR-146a-/- protection from MCMV. Surprisingly, NK cells from miR-146a-deficient mice were largely similar to control NK cells with respect to development, maturation, trafficking, and effector functions. However, miR-146a-/- mice had increased NK cell numbers and frequency of the most mature Stage IV (CD27-CD11b+) NK cells in the liver at baseline, enhanced STAT1 phosphorylation, and increased selective expansion of Ly49H+ NK cells and T cells during MCMV infection. This study demonstrates a critical role for miR-146a in the host response to MCMV, arising from mechanisms that include increased NK cell numbers and early T-cell expansion.

Natural killer cell effector function is critical for host defense against alcohol-associated bacterial pneumonia.

Alcohol use is an independent risk factor for the development of bacterial pneumonia due, in part, to impaired mucus-facilitated clearance, macrophage phagocytosis, and recruitment of neutrophils. Alcohol consumption is also known to reduce peripheral natural killer (NK) cell numbers and compromise NK cell cytolytic activity, especially NK cells with a mature phenotype. However, the role of innate lymphocytes, such as NK cells during host defense against alcohol-associated bacterial pneumonia is essentially unknown. We have previously shown that indole supplementation mitigates increases in pulmonary bacterial burden and improves pulmonary NK cell recruitment in alcohol-fed mice, which were dependent on aryl hydrocarbon receptor (AhR) signaling. Employing a binge-on-chronic alcohol-feeding model we sought to define the role and interaction of indole and NK cells during pulmonary host defense against alcohol-associated pneumonia. We demonstrate that alcohol dysregulates NK cell effector function and pulmonary recruitment via alterations in two key signaling pathways. We found that alcohol increases transforming growth factor beta (TGF-ß) signaling while suppressing AhR signaling. We further demonstrated that NK cells isolated from alcohol-fed mice have a reduced ability to kill Klebsiella pneumoniae. NK cell migratory capacity to chemokines was also significantly altered by alcohol, as NK cells isolated from alcohol-fed mice exhibited preferential migration in response to CXCR3 chemokines but exhibited reduced migration in response to CCR2, CXCR4, and CX3CR1 chemokines. Together this data suggests that alcohol disrupts NK cell-specific TGF-ß and AhR signaling pathways leading to decreased pulmonary recruitment and cytolytic activity thereby increasing susceptibility to alcohol-associated bacterial pneumonia.

Phase II Study of Samotolisib in Children and Young Adults With Tumors Harboring Phosphoinositide 3-Kinase/Mammalian Target of Rapamycin Pathway Alterations: Pediatric MATCH APEC1621D.

PURPOSE: Patients age 1-21 years with relapsed or refractory solid and CNS tumors were assigned to phase II studies of molecularly targeted therapies on the National Cancer Institute-Children's Oncology Group (NCI-COG) Pediatric Molecular Analysis for Therapy Choice (MATCH) trial. Patients whose tumors harbored predefined genetic alterations in the phosphoinositide 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway and lacked mitogen-activated protein kinase pathway activating alterations were treated with the PI3K/mTOR inhibitor samotolisib. METHODS: Patients received samotolisib twice daily in 28-day cycles until disease progression or unacceptable toxicity. A rolling 6 limited dose escalation was performed as, to our knowledge, this was the first pediatric study of samotolisib. The primary end point was the objective response rate; secondary end points included progression-free survival (PFS) and the recommended phase II dose and toxicity of samotolisib in children. RESULTS: A total of 3.4% (41/1,206) of centrally tested patients were matched to this arm. Seventeen patients were treated. Among treated patients, the most common diagnoses included osteosarcoma (n = 6) and high-grade glioma (n = 5) harboring alterations in phosphatase and tensin homolog (n = 6), PIK3CA (n = 5), and tuberous sclerosis complex 2 (n = 3). No objective responses or prolonged stable disease were observed. Three-month PFS was 12% (95% CI, 2 to 31). Two patients experienced dose-limiting toxicities (mucositis and pneumonitis). Dose level 2 (115 mg/m2/dose twice daily) was determined to be the recommended phase II dose of samotolisib in children. CONCLUSION: This nationwide study was successful at identifying patients and evaluating the efficacy of molecularly targeted therapy for rare molecular subgroups of patients in a histology-agnostic fashion. Unfortunately, there was no activity of samotolisib against tumors with PI3K/mTOR pathway alterations. Prospective trials such as the NCI-COG Pediatric MATCH are necessary to evaluate the efficacy of molecularly targeted therapies given their increasing use in clinical practice.

Population-Based Estimates of the Prevalence of Children With Congenital Heart Disease and Associated Comorbidities in the United States.

BACKGROUND: Congenital heart defects (CHD) are the most common birth defects and previous estimates report the disease affects 1% of births annually in the United States. To date, CHD prevalence estimates are inconsistent due to varied definitions, data reliant on birth registries, and are geographically limited. These data sources may not be representative of the total prevalence of the CHD population. It is therefore important to derive high-quality, population-based estimates of the prevalence of CHD to help care for this vulnerable population. METHODS: We performed a descriptive, retrospective 8-year analysis using all-payer claims data from Colorado from 2012 to 2019. Children with CHD were identified by applying International Classification of Diseases-Ninth Revision (ICD-9) and International Classification of Diseases-Tenth Revision (ICD-10) diagnosis codes from the American Heart Association-American College of Cardiology harmonized cardiac codes. We included children with CHD <18 years of age who resided in Colorado, had a documented zip code, and had at least 1 health care claim. CHD type was categorized as simple, moderate, and severe disease. Association with comorbid conditions and genetic diagnoses were analyzed using χ2 test. We used direct standardization to calculate adjusted prevalence rates, controlling for age, sex, primary insurance provider, and urban-rural residence. RESULTS: We identified 1 566 328 children receiving care in Colorado from 2012 to 2019. Of those, 30 512 children had at least 1 CHD diagnosis, comprising 1.95% (95% CI, 1.93-1.97) of the pediatric population. Over half of the children with CHD also had at least 1 complex chronic condition. After direct standardization, the adjusted prevalence rates show a small increase in simple severity diagnoses across the study period (adjusted rate of 11.5 [2012]-14.4 [2019]; P<0.001). CONCLUSIONS: The current study is the first population-level analysis of pediatric CHD in the United States. Using administrative claims data, our study found a higher CHD prevalence and comorbidity burden compared with previous estimates.