Hyperferritinemia screening to aid identification and differentiation of patients with hyperinflammatory disorders.

High ferritin is an important and sensitive biomarker for hemophagocytic lymphohistiocytosis (HLH), a diverse and deadly group of cytokine storm syndromes. Early action to prevent immunopathology in HLH often includes empiric immunomodulation, which can complicate etiologic work-up and prevent collection of early/pre-treatment research samples. To address this, we instituted an alert system where serum ferritin > 1000ng/mL triggered real-time chart review, assessment of whether the value reflected "inflammatory hyperferritnemia (IHF)", and biobanking of remnant samples from consenting IHF patients. We extracted relevant clinical data; periodically measured serum total IL-18, IL-18 binding protein (IL-18BP), and CXCL9; retrospectively classified patients by etiology into infectious, rheumatic, or immune dysregulation; and subjected a subgroup of samples to a 96-analyte biomarker screen. 180 patients were identified, 30.5% of which had IHF. Maximum ferritin levels were significantly higher in patients with IHF than with either hemoglobinopathy or transplant, and highly elevated total IL-18 levels were distinctive to patients with Stills Disease and/or Macrophage Activation Syndrome (MAS). Multi-analyte analysis showed elevation in proteins associated with cytotoxic lymphocytes in all IHF samples when compared to healthy controls and depression of proteins such as ANGPT1 and VEGFR2 in samples from hyperferritinemic sepsis patients relative to non-sepsis controls. This single-center, real-time IFH screen proved feasible and efficient, validated prior observations about the specificity of IL-18, enabled early sample collection from a complex population, suggested a unique vascular biomarker signature in hyperferritinemic sepsis, and expanded our understanding of IHF heterogeneity.

The Effects of TMR® Fab 6 on Hamstring Flexibility in Healthy Subjects; An Exploratory Observational Investigation.

Background: Stretching programs are designed to improve hamstring flexibility by attempting to mechanically increase the length of the target tissue. However, other manual treatment approaches such as those utilized in Total Motion Release (TMR®), could be beneficial by identifying body asymmetries to assess and treat soft tissue impairments leading to diminished extensibility. Purpose: The purpose of this study was to determine the effectiveness of the TMR® Fab 6 assessment and treatment to increase hamstring flexibility in healthy participants following one session of TMR®. Study Design: Observational Cohort study. Methods: A convenience sample of 20 healthy participants (10 males, 10 females) were recruited from three institutions. Following collection of demographic information and a brief medical history, each participant performed a five minute warm-up on the stationary bike at a moderate intensity (80-90 RPMs) followed immediately by the bilateral performance of the Active Knee Extension Test (AKET) and Passive Straight Leg Raise (PSLR) to assess hamstring muscle length. Participants were randomly placed in the TMR® or control group. The TMR® group completed the "Fab 6" evaluation and treatment, while the control group performed one repetition of standing active hip flexion every 30-seconds for 15-minutes with both knees in full extension. Upon completion of treatment, control and TMR® groups were immediately re-evaluated on the AKET and the PSLR in the same order and fashion as baseline testing. Participants were asked to return in 24-hours for the same objective measurements as previously described. Results: A significant time by group interaction was identified across all variables (p ≤ 0.001) for AKET and PSLR except the PSLR preferred leg from post-treatment to 24hr follow-up. The most significant increase in the AKET occurred in the TMR® group between baseline and post-treatment of the non-preferred leg (12.15°±2.94) when compared to the control group (7.15°±1.56). Conclusion: The results of the study suggest that implementing a regionally interdependent treatment approach like TMR® results in significant improvements in hamstring extensibility and hip ROM compared to the control group. Level of evidence: 3.

Mechanisms of resistance to oncogenic KRAS inhibition in pancreatic cancer.

KRAS inhibitors demonstrate clinical efficacy in pancreatic ductal adenocarcinoma (PDAC); however, resistance is common. Among patients with KRASG12C-mutant PDAC treated with adagrasib or sotorasib, mutations in PIK3CA and KRAS, and amplifications of KRASG12C, MYC, MET, EGFR, and CDK6 emerged at acquired resistance. In PDAC cell lines and organoid models treated with the KRASG12D inhibitor MRTX1133, epithelial-to-mesenchymal transition and PI3K-AKT-mTOR signaling associate with resistance to therapy. MRTX1133 treatment of the KrasLSL-G12D/+;Trp53LSL-R172H/+;p48-Cre (KPC) mouse model yielded deep tumor regressions, but drug resistance ultimately emerged, accompanied by amplifications of Kras, Yap1, Myc, and Cdk6/Abcb1a/b, and co-evolution of drug-resistant transcriptional programs. Moreover, in KPC and PDX models, mesenchymal and basal-like cell states displayed increased response to KRAS inhibition compared to the classical state. Combination treatment with KRASG12D inhibition and chemotherapy significantly improved tumor control in PDAC mouse models. Collectively, these data elucidate co-evolving resistance mechanisms to KRAS inhibition and support multiple combination therapy strategies.

Sex and Sexual Orientation Differences in Dark Triad Traits, Sexual Excitation/Inhibition, and Sociosexuality.

The present study sought to investigate sex and sexual orientation differences in several traits related to sexuality and sexual behavior. Examining sexual orientation differences alongside basic sex differences to help identify correlates of sexual orientation diversity, and whether individuals with varying degrees of same-sex attraction show concurrent sex-atypical shifts in other domains. Males tend to score higher than females in the Dark Triad (DT) traits of sub-clinical narcissism, psychopathy, and Machiavellianism. Similarly, females tend to be more cautious than males in their attitudes and desires toward casual sex activity (i.e., sociosexuality). These sex differences may be related to the propensity for individuals to become easily sexually excited, which is higher in males, or to instead inhibit sexual arousal, which is higher in females. In a large undergraduate sample (N = 2047), we replicated expected sex differences in DT traits, sociosexuality, and sexual excitation/inhibition. We found that non-heterosexual females were "male-shifted" in some of these traits, but these shifts tended to be strongest among mostly heterosexual and bisexual individuals. Furthermore, we found that within-sex variation in sociosexuality, sexual excitation, and sexual inhibition was not related to sexual orientation in a linear fashion. Instead, sociosexuality and sexual excitation were related to sexual orientation in a curvilinear (inverted-U) fashion, especially among females. The fact that traits correlated with bisexuality and homosexuality were somewhat distinct is consistent with the idea that different developmental pathways may lead to these discrete sexual attraction patterns.

Correlation of limited-early-response status with 12-month CST, BVA, and machine learning-quantified retinal fluid in diabetic macular oedema in routine clinical practice.

BACKGROUND/OBJECTIVES: Anti-VEGF treatment response in DMO has been measured by changes in the central subfield thickness (CST) and best visual acuity (BVA) outcomes at 3 months after initial treatment, termed early or limited early response (ER/LER). This study correlates LER with 12-month BVA, CST, and retinal fluid volumes quantified by a machine learning algorithm on optical coherence tomography (OCT). SUBJECTS/METHODS: The study included treatment naïve DMO patients ≥ 18 years with OCT scans at baseline (M0), M3, M6, and M12. The 220 patients were categorized as limited early responders (LER) if they had ≤ 10% CST reduction and/or < 5 ETDRS letter gain at M3. BVA, CST, and subretinal (SRF), intraretinal (IRF), and total retinal (TRF) fluid volumes quantified by a machine learning algorithm were compared between groups and across time. RESULTS: At M12, the anatomic LER (aLER), defined solely by CST, had significantly worse BVA and CST versus the anatomic ER (aER) group (p < 0.001). Retinal fluid M12 outcomes did not significantly vary between all LER and ER groups. No significant BVA, CST, TRF, and IRF variance across time for LER was found (p > 0.1). CONCLUSIONS: BVA and CST M12 outcomes vary by aLER/aER status indicating that CST may be a strong predictor of treatment outcomes, while retinal fluid volumes were not predicted by LER status.

Association of Glomerular Filtration Rate Decline With Clinical Outcomes in a Population With Type 2 Diabetes.

Background: While historical rate of decline in kidney function is informally used by clinicians to estimate risk of future adverse clinical outcomes especially kidney failure, in people with type 2 diabetes the epidemiology and independent association of historical eGFR slope on risk is not well described. Objective: Determine the association of eGFR slope and risk of clinically important outcomes. Design Setting and Patients: Observational population-based cohort with type 2 diabetes in Alberta. Measurement and Methods: An Alberta population-based cohort with type 2 diabetes was assembled, characterized, and observed over 1 year (2018) for clinical outcomes of ESKD, first myocardial infarction, first stroke, heart failure, and disease-specific and all-cause hospitalization and mortality. Kidney function was defined using KDIGO criteria using the most recent eGFR and albuminuria measured in the preceding 18 months; annual eGFR slope utilized measurements in the 3 years prior and was parameterized using three methods (percentiles, and linear term with and without missingness indicator). Demographics, laboratory results, medications, and comorbid conditions using validated definitions were described. In addition to descriptive analysis, odds ratios from fully adjusted logistic models regressing outcomes on eGFR slope are reported; the marginal risk of clinical outcomes was also determined. Results: Among 336 376 participants with type 2 diabetes, the median annual eGFR slope was -0.41 mL/min/1.73 m2 (IQR -1.67, 0.62). In fully adjusted models, eGFR slope was independently associated with many adverse clinical outcomes; among those with ≤10th percentile of slope (median -4.71 mL/min/1.73 m2) the OR of kidney failure was 2.22 (95% CI 1.75, 2.82), new stroke 1.23 (1.08, 1.40), heart failure 1.42 (1.27, 1.59), MI 0.98 (0.77, 1.23) all-cause hospitalization 1.31 (1.26, 1.36) and all-cause mortality 1.56 (1.44, 1.68). For every -1 mL/min/1.73 m2 in eGFR slope, the OR of outcomes ranged from 1.01 (0.98, 1.05 for new MI) to 1.09 (1.08, 1.10 for all-cause mortality); findings were significant for 10 of the 13 outcomes considered. Limitations: Causality cannot be established with this study design. Conclusions: These findings support consideration of the rate of eGFR decline in risk stratification and may inform clinicians and policymakers to optimize treatment and inform health care system planning.

Contexte: Bien que les antécédents de déclin de la fonction rénale soient utilisés de manière informelle par les cliniciens pour estimer le risque d'issues cliniques défavorables ­ particulièrement l'insuffisance rénale terminale (IRT) ­ chez les diabétiques de type 2, l'épidémiologie de la pente du DFGe et son association indépendante sur ce risque demeurent mal décrites. Objectif: Examiner l'association entre la pente du DFGe et le risque de résultats d'importance clinique. Sujets et conception de l'étude: Étude de cohorte observationnelle basée sur une population d'Albertains atteints de diabète de type 2. Méthodologie et mesures: Nous avons constitué, caractérisé et observé une cohorte d'Albertains atteints de diabète de type 2 sur une période d'un an (2018) pour les résultats cliniques suivants: IRT, premier infarctus du myocarde (IM), premier AVC, insuffisance cardiaque, ainsi que les hospitalisations et la mortalité liées à la maladie et à toutes causes confondues. La fonction rénale a été définie selon les critères KDIGO à partir des plus récentes valeurs de DFGe et d'albuminurie mesurées dans les 18 mois précédents. La pente annuelle du DFGe a été calculée à partir des mesures effectuées au cours des trois années précédentes et paramétrée selon trois méthodes (percentiles, termes linéaires avec et sans indications de données manquantes). Les données démographiques, les résultats de laboratoire, les médicaments et les comorbidités ont été décrits selon les définitions validées. En plus de l'analyze descriptive, des rapports de cotes (RC) pour les résultats liés au déclin du DFGe ont été établis à l'aide de modèles de régression logistique entièrement ajustés; le risque marginal des résultats cliniques d'intérêt a également été déterminé. Résultats: Parmi les 336 376 diabétiques de type 2 participants, la pente annuelle médiane du DFGe s'établissait à −0,41 ml/min/1,73 m2 (ÉIQ: −1,67 à 0,62). Dans les modèles ajustés, la pente du DFGe a été associée de façon indépendante à plusieurs issues cliniques défavorables. Parmi ceux qui présentaient une pente du DFGe ≤10e percentile (médiane: −4,71 ml/min/1,73 m2), le RC était de 2,22 (IC 95 %: 1,75 à 2,82) pour l'insuffisance rénale; de 1,23 (1,08 à 1,40) pour les nouveaux AVC; de 1,42 (1,27 à 1,59) pour l'insuffisance cardiaque; de 0,98 (0,77 à 1,23) pour les nouveaux IM; de 1,31 (1,26 à 1,36) pour les hospitalisations toutes causes confondues et de 1,56 (1,44 à 1,68) pour la mortalité toutes causes confondues. Pour chaque tranche de - 1 ml/min/1,73 m2 de la pente du DFGe, le RC des résultats cliniques variait de 1,01 (0,98 à 1,05) pour les nouveaux IM à 1,09 (1,08 à 1,10) pour la mortalité toutes causes confondues; les résultats étaient significatifs pour 10 des 13 résultats examinés. Limites: La causalité ne peut pas être établie avec ce plan d'étude. Conclusion: Ces résultats plaident en faveur de la prise en compte du taux de déclin du DFGe dans la stratification du risque. Ils peuvent également aider les cliniciens et les décideurs à optimiser le traitement et à planifier les systèmes de soins de santé.

Intrinsic Dynamics and Neural Implementation of a Hypothalamic Line Attractor Encoding an Internal Behavioral State.

Line attractors are emergent population dynamics hypothesized to encode continuous variables such as head direction and internal states. In mammals, direct evidence of neural implementation of a line attractor has been hindered by the challenge of targeting perturbations to specific neurons within contributing ensembles. Estrogen receptor type 1 (Esr1)-expressing neurons in the ventrolateral subdivision of the ventromedial hypothalamus (VMHvl) show line attractor dynamics in male mice during fighting. We hypothesized that these dynamics may encode continuous variation in the intensity of an internal aggressive state. Here, we report that these neurons also show line attractor dynamics in head-fixed mice observing aggression. We exploit this finding to identify and perturb line attractor-contributing neurons using 2-photon calcium imaging and holographic optogenetic perturbations. On-manifold perturbations demonstrate that integration and persistent activity are intrinsic properties of these neurons which drive the system along the line attractor, while transient off-manifold perturbations reveal rapid relaxation back into the attractor. Furthermore, stimulation and imaging reveal selective functional connectivity among attractor-contributing neurons. Intriguingly, individual differences among mice in line attractor stability were correlated with the degree of functional connectivity among contributing neurons. Mechanistic modelling indicates that dense subnetwork connectivity and slow neurotransmission are required to explain our empirical findings. Our work bridges circuit and manifold paradigms, shedding light on the intrinsic and operational dynamics of a behaviorally relevant mammalian line attractor.

The hidden professionalism curriculum: Teach it, see it, do it and repeat!

Background: Professionalism is a complex and multifaceted component of medical education. Historically, students have learned about professionalism informally and as part of the hidden curriculum. Currently, professionalism is increasingly prominent in formal curricula, but uncertainty remains regarding optimal professionalism pedagogies. In this study, the authors explored medical students' exposure to professional topics and considered factors that enabled students to correctly recognize and manage these issues. Methods: Convenience sampling was used to recruit medical students from existing clinical attachments at the authors' hospital. A semi-structured interview format was used to explore participants' awareness of professional issues within fictional vignettes created using published regulatory guidance. The interview transcripts and interview guide field notes were then analyzed. Results: The data suggest that students require a combination of didactic teaching and experiential learning to reliably recognize and manage professional issues. Didactic teaching alone enabled topic recognition, but with uncertainty about management strategies. Experiential learning alone led to erratic recognition of the subject and reliance upon role modeling to guide its management. This work stimulates faculty development to enhance teaching professionalism. Conclusions: Undergraduate medical education on professionalism must be introduced into the formal curriculum. Didactic teaching is required to scaffold experiential learning. Failure to do so renders students unable to reliably recognize or manage professional issues encountered in clinical practice. Further research questions were identified to progress this work.

Physical Activity and Neuropsychological Functioning in Older Adults.

Research has identified a positive relationship between physical activity and neuropsychological functioning across the lifespan. The present study further examined the relationship between physical activity, depression, anxiety, and cognitive functioning in community-dwelling older adults (ages 65-96) who completed an outpatient neuropsychological evaluation (N = 526). Psychometrically sound and validated measures were used to assess depression, anxiety, and cognitive functioning. Analyses of covariance (ANCOVA) were conducted to examine differences between individuals who reported regularly engaging in physical activity and those who did not, after adjusting for demographic variables (age, education, and gender). Results indicated that physical activity was associated with better scores on measures of depression, anxiety, and cognitive functioning. Effect sizes for total scores on all measures were large, but there was a sizeable range of effect sizes (from small to large) for various cognitive domains. Smallest effect sizes were observed for subtests measuring language skills and visuospatial abilities, whereas largest effect sizes were seen in processing speed and memory. Results suggest that engaging in physical activity may be a beneficial non-pharmacological intervention for older adults. These findings underscore the importance of integrating physical activity programs in community and healthcare settings to foster mental and cognitive health in older populations.

Nomenclature of the radial nerve: distinguishing between the deep branch of the radial nerve and the posterior interosseous nerve.

To address an inconsistency in the nomenclature of the anatomy and compressive syndromes of the deep branch of the radial nerve, we advocate for a single compression syndrome that presents along a spectrum from pain to posterior interosseous nerve palsy.

The Complex Latent Structure of Attenuated Psychotic Symptoms: Hierarchical and Bifactor Models of SIPS Symptoms Replicated in Two Large Samples at Clinical High Risk for Psychosis.

BACKGROUND AND HYPOTHESIS: The Structured Interview for Psychosis-Risk Syndromes (SIPS) and other assessments of psychosis risk define clinical high risk for psychosis (CHR) by the presence of attenuated psychotic symptoms. Despite extensive research on attenuated psychotic symptoms, substantial questions remain about their internal psychometric structure and relationships to comorbid non-psychotic symptoms. STUDY DESIGN: Hierarchical and bifactor models were developed for the SIPS in a large CHR sample (NAPLS-3, N = 787) and confirmed through preregistered replication in an independent sample (NAPLS-2, N = 1043). Criterion validity was tested through relationships with CHR status, comorbid symptoms/diagnoses, functional impairment, demographics, neurocognition, and conversion to psychotic disorders. STUDY RESULTS: Most variance in SIPS items (75%-77%) was attributable to a general factor. Hierarchical and bifactor models included a general factor and five specific/lower-order factors (positive symptoms, eccentricity, avolition, lack of emotion, and deteriorated thought process). CHR participants were elevated on the general factor and the positive symptoms factor. The general factor was associated with depressive symptoms; functional impairment; and mood, anxiety, and schizotypal personality diagnoses. The general factor was the best predictor of psychotic disorders (d ≥ 0.50). Positive symptoms and eccentricity had specific effects on conversion outcomes. The deteriorated thought process was least meaningful/replicable. CONCLUSIONS: Attenuated psychotic symptoms, measured by the SIPS, have a complex hierarchical structure with a strong general factor. The general factor relates to internalizing symptoms and functional impairment, emphasizing the roles of general psychopathological distress/impairment in psychosis risk. Shared symptom variance complicates the interpretation of raw symptom scores. Broad transdiagnostic assessment is warranted to model psychosis risk accurately.

An age-progressive platelet differentiation path from hematopoietic stem cells causes exacerbated thrombosis.

Platelet dysregulation is drastically increased with advanced age and contributes to making cardiovascular disorders the leading cause of death of elderly humans. Here, we reveal a direct differentiation pathway from hematopoietic stem cells into platelets that is progressively propagated upon aging. Remarkably, the aging-enriched platelet path is decoupled from all other hematopoietic lineages, including erythropoiesis, and operates as an additional layer in parallel with canonical platelet production. This results in two molecularly and functionally distinct populations of megakaryocyte progenitors. The age-induced megakaryocyte progenitors have a profoundly enhanced capacity to engraft, expand, restore, and reconstitute platelets in situ and upon transplantation and produce an additional platelet population in old mice. The two pools of co-existing platelets cause age-related thrombocytosis and dramatically increased thrombosis in vivo. Strikingly, aging-enriched platelets are functionally hyper-reactive compared with the canonical platelet populations. These findings reveal stem cell-based aging as a mechanism for platelet dysregulation and age-induced thrombosis.

Nematode histone H2A variant evolution reveals diverse histories of retention and loss and evidence for conserved core-like variant histone genes.

Histone variants are paralogs that replace canonical histones in nucleosomes, often imparting novel functions. However, how histone variants arise and evolve is poorly understood. Reconstruction of histone protein evolution is challenging due to large differences in evolutionary rates across gene lineages and sites. Here we used intron position data from 108 nematode genomes in combination with amino acid sequence data to find disparate evolutionary histories of the three H2A variants found in Caenorhabditis elegans: the ancient H2A.ZHTZ-1, the sperm-specific HTAS-1, and HIS-35, which differs from the canonical S-phase H2A by a single glycine-to-alanine C-terminal change. Although the H2A.ZHTZ-1 protein sequence is highly conserved, its gene exhibits recurrent intron gain and loss. This pattern suggests that specific intron sequences or positions may not be important to H2A.Z functionality. For HTAS-1 and HIS-35, we find variant-specific intron positions that are conserved across species. Patterns of intron position conservation indicate that the sperm-specific variant HTAS-1 arose more recently in the ancestor of a subset of Caenorhabditis species, while HIS-35 arose in the ancestor of Caenorhabditis and its sister group, including the genus Diploscapter. HIS-35 exhibits gene retention in some descendent lineages but gene loss in others, suggesting that histone variant use or functionality can be highly flexible. Surprisingly, we find the single amino acid differentiating HIS-35 from core H2A is ancestral and common across canonical Caenorhabditis H2A sequences. Thus, we speculate that the role of HIS-35 lies not in encoding a functionally distinct protein, but instead in enabling H2A expression across the cell cycle or in distinct tissues. This work illustrates how genes encoding such partially-redundant functions may be advantageous yet relatively replaceable over evolutionary timescales, consistent with the patchwork pattern of retention and loss of both genes. Our study shows the utility of intron positions for reconstructing evolutionary histories of gene families, particularly those undergoing idiosyncratic sequence evolution.

Associations of street-view greenspace with Parkinson's disease hospitalizations in an open cohort of elderly US Medicare beneficiaries.

INTRODUCTION: Protective associations of greenspace with Parkinson's disease (PD) have been observed in some studies. Visual exposure to greenspace seems to be important for some of the proposed pathways underlying these associations. However, most studies use overhead-view measures (e.g., satellite imagery, land-classification data) that do not capture street-view greenspace and cannot distinguish between specific greenspace types. We aimed to evaluate associations of street-view greenspace measures with hospitalizations with a PD diagnosis code (PD-involved hospitalization). METHODS: We created an open cohort of about 45.6 million Medicare fee-for-service beneficiaries aged 65 + years living in core based statistical areas (i.e. non-rural areas) in the contiguous US (2007-2016). We obtained 350 million Google Street View images across the US and applied deep learning algorithms to identify percentages of specific greenspace features in each image, including trees, grass, and other green features (i.e., plants, flowers, fields). We assessed yearly average street-view greenspace features for each ZIP code. A Cox-equivalent re-parameterized Poisson model adjusted for potential confounders (i.e. age, race/ethnicity, socioeconomic status) was used to evaluate associations with first PD-involved hospitalization. RESULTS: There were 506,899 first PD-involved hospitalizations over 254,917,192 person-years of follow-up. We found a hazard ratio (95% confidence interval) of 0.96 (0.95, 0.96) per interquartile range (IQR) increase for trees and a HR of 0.97 (0.96, 0.97) per IQR increase for other green features. In contrast, we found a HR of 1.06 (1.04, 1.07) per IQR increase for grass. Associations of trees were generally stronger for low-income (i.e. Medicaid eligible) individuals, Black individuals, and in areas with a lower median household income and a higher population density. CONCLUSION: Increasing exposure to trees and other green features may reduce PD-involved hospitalizations, while increasing exposure to grass may increase hospitalizations. The protective associations may be stronger for marginalized individuals and individuals living in densely populated areas.

Structural and molecular basis of choline uptake into the brain by FLVCR2.

Choline is an essential nutrient that the human body needs in vast quantities for cell membrane synthesis, epigenetic modification and neurotransmission. The brain has a particularly high demand for choline, but how it enters the brain remains unknown1-3. The major facilitator superfamily transporter FLVCR1 (also known as MFSD7B or SLC49A1) was recently determined to be a choline transporter but is not highly expressed at the blood-brain barrier, whereas the related protein FLVCR2 (also known as MFSD7C or SLC49A2) is expressed in endothelial cells at the blood-brain barrier4-7. Previous studies have shown that mutations in human Flvcr2 cause cerebral vascular abnormalities, hydrocephalus and embryonic lethality, but the physiological role of FLVCR2 is unknown4,5. Here we demonstrate both in vivo and in vitro that FLVCR2 is a BBB choline transporter and is responsible for the majority of choline uptake into the brain. We also determine the structures of choline-bound FLVCR2 in both inward-facing and outward-facing states using cryo-electron microscopy. These results reveal how the brain obtains choline and provide molecular-level insights into how FLVCR2 binds choline in an aromatic cage and mediates its uptake. Our work could provide a novel framework for the targeted delivery of therapeutic agents into the brain.

Juvenile survival increases with dispersal distance and varies across years: 15 years of evidence in a prairie perennial.

Juvenile survival is critical to population persistence and evolutionary change. However, the survival of juvenile plants from emergence to reproductive maturity is rarely quantified. This is especially true for long-lived perennials with extended pre-reproductive periods. Furthermore, studies rarely have the replication necessary to account for variation among populations and cohorts. We estimated juvenile survival and its relationship to population size, density of conspecifics, distance to the maternal plant, age, year, and cohort for Echinacea angustifolia, a long-lived herbaceous perennial. In 14 remnant prairie populations over seven sampling years, 2007-2013, we identified 886 seedlings. We then monitored these individuals annually until 2021 (8-15 years). Overall, juvenile mortality was very high; for almost all cohorts fewer than 10% of seedlings survived to age 8 or to year 2021. Only two of the seedlings reached reproductive maturity within the study period. Juvenile survival increased with distance from the maternal plant and varied more among the study years than it did by age or cohort. Juvenile survival did not vary with population size or local density of conspecific neighbors. Our results suggest that low juvenile survival could contribute to projected population declines.

The state of the art in secondary pharmacology and its impact on the safety of new medicines.

Secondary pharmacology screening of investigational small-molecule drugs for potentially adverse off-target activities has become standard practice in pharmaceutical research and development, and regulatory agencies are increasingly requesting data on activity against targets with recognized adverse effect relationships. However, the screening strategies and target panels used by pharmaceutical companies may vary substantially. To help identify commonalities and differences, as well as to highlight opportunities for further optimization of secondary pharmacology assessment, we conducted a broad-ranging survey across 18 companies under the auspices of the DruSafe leadership group of the International Consortium for Innovation and Quality in Pharmaceutical Development. Based on our analysis of this survey and discussions and additional research within the group, we present here an overview of the current state of the art in secondary pharmacology screening. We discuss best practices, including additional safety-associated targets not covered by most current screening panels, and present approaches for interpreting and reporting off-target activities. We also provide an assessment of the safety impact of secondary pharmacology screening, and a perspective on opportunities and challenges in this rapidly developing field.

Preclinical evaluation of MC1R targeting theranostic pair [155Tb]Tb-crown-αMSH and [161Tb]Tb-crown-αMSH.

BACKGROUND: Targeted radionuclide therapy is established as a highly effective strategy for the treatment of metastatic tumors; however, the co-development of suitable imaging companions to therapy remains significant challenge. Theranostic isotopes of terbium (149Tb, 152Tb, 155Tb, 161Tb) have the potential to provide chemically identical radionuclidic pairs, which collectively encompass all modes of nuclear decay relevant to nuclear medicine. Herein, we report the first radiochemistry and preclinical studies involving 155Tb- and 161Tb-labeled crown-αMSH, a small peptide-based bioconjugate suitable for targeting melanoma. METHODS: 155Tb was produced via proton induced spallation of Ta targets using the isotope separation and acceleration facility at TRIUMF with isotope separation on-line (ISAC/ISOL). The radiolabeling characteristics of crown-αMSH with 155Tb and/or 161Tb were evaluated by concentration-dependence radiolabeling studies, and radio-HPLC stability studies. LogD7.4 measurements were obtained for [161Tb]Tb-crown-αMSH. Competitive binding assays were undertaken to determine the inhibition constant for [natTb]Tb-crown-αMSH in B16-F10 cells. Pre-clinical biodistribution and SPECT/CT imaging studies of 155Tb and 161Tb labeled crown-αMSH were undertaken in male C57Bl/6 J mice bearing B16-F10 melanoma tumors to evaluate tumor specific uptake and imaging potential for each radionuclide. RESULTS: Quantitative radiolabeling of crown-αMSH with [155Tb]Tb3+ and [161Tb]Tb3+ was demonstrated under mild conditions (RT, 10 min) and low chelator concentrations; achieving high molar activities (23-29 MBq/nmol). Radio-HPLC studies showed [161Tb]Tb-crown-αMSH maintains excellent radiochemical purity in human serum, while gradual metabolic degradation is observed in mouse serum. Competitive binding assays showed the high affinity of [natTb]Tb-crown-αMSH toward MC1R. Two different methods for preparation of the [155Tb]Tb-crown-αMSH radiotracer were investigated and the impacts on the biodistribution profile in tumor bearing mice is compared. Preclinical in vivo studies of 155Tb- and 161Tb- labeled crown-αMSH were performed in parallel, in mice bearing B16-F10 tumors; where the biodistribution results showed similar tumor specific uptake (6.06-7.44 %IA/g at 2 h pi) and very low uptake in nontarget organs. These results were further corroborated through a series of single-photon emission computed tomography (SPECT) studies, with [155Tb]Tb-crown-αMSH and [161Tb]Tb-crown-αMSH showing comparable uptake profiles and excellent image contrast. CONCLUSIONS: Collectively, our studies highlight the promising characteristics of [155Tb]Tb-crown-αMSH and [161Tb]Tb-crown-αMSH as theranostic pair for nuclear imaging (155Tb) and radionuclide therapy (161Tb).