p53 governs an AT1 differentiation programme in lung cancer suppression.

Lung cancer is the leading cause of cancer deaths worldwide1. Mutations in the tumour suppressor gene TP53 occur in 50% of lung adenocarcinomas (LUADs) and are linked to poor prognosis1-4, but how p53 suppresses LUAD development remains enigmatic. We show here that p53 suppresses LUAD by governing cell state, specifically by promoting alveolar type 1 (AT1) differentiation. Using mice that express oncogenic Kras and null, wild-type or hypermorphic Trp53 alleles in alveolar type 2 (AT2) cells, we observed graded effects of p53 on LUAD initiation and progression. RNA sequencing and ATAC sequencing of LUAD cells uncovered a p53-induced AT1 differentiation programme during tumour suppression in vivo through direct DNA binding, chromatin remodelling and induction of genes characteristic of AT1 cells. Single-cell transcriptomics analyses revealed that during LUAD evolution, p53 promotes AT1 differentiation through action in a transitional cell state analogous to a transient intermediary seen during AT2-to-AT1 cell differentiation in alveolar injury repair. Notably, p53 inactivation results in the inappropriate persistence of these transitional cancer cells accompanied by upregulated growth signalling and divergence from lung lineage identity, characteristics associated with LUAD progression. Analysis of Trp53 wild-type and Trp53-null mice showed that p53 also directs alveolar regeneration after injury by regulating AT2 cell self-renewal and promoting transitional cell differentiation into AT1 cells. Collectively, these findings illuminate mechanisms of p53-mediated LUAD suppression, in which p53 governs alveolar differentiation, and suggest that tumour suppression reflects a fundamental role of p53 in orchestrating tissue repair after injury.

Innate immune cell activation causes lung fibrosis in a humanized model of long COVID.

COVID-19 remains a global pandemic of an unprecedented magnitude with millions of people now developing "COVID lung fibrosis." Single-cell transcriptomics of lungs of patients with long COVID revealed a unique immune signature demonstrating the upregulation of key proinflammatory and innate immune effector genes CD47, IL-6, and JUN. We modeled the transition to lung fibrosis after COVID and profiled the immune response with single-cell mass cytometry in JUN mice. These studies revealed that COVID mediated chronic immune activation reminiscent to long COVID in humans. It was characterized by increased CD47, IL-6, and phospho-JUN (pJUN) expression which correlated with disease severity and pathogenic fibroblast populations. When we subsequently treated a humanized COVID lung fibrosis model by combined blockade of inflammation and fibrosis, we not only ameliorated fibrosis but also restored innate immune equilibrium indicating possible implications for clinical management of COVID lung fibrosis in patients.

Identification of cell types in multiplexed in situ images by combining protein expression and spatial information using CELESTA.

Advances in multiplexed in situ imaging are revealing important insights in spatial biology. However, cell type identification remains a major challenge in imaging analysis, with most existing methods involving substantial manual assessment and subjective decisions for thousands of cells. We developed an unsupervised machine learning algorithm, CELESTA, which identifies the cell type of each cell, individually, using the cell's marker expression profile and, when needed, its spatial information. We demonstrate the performance of CELESTA on multiplexed immunofluorescence images of colorectal cancer and head and neck squamous cell carcinoma (HNSCC). Using the cell types identified by CELESTA, we identify tissue architecture associated with lymph node metastasis in HNSCC, and validate our findings in an independent cohort. By coupling our spatial analysis with single-cell RNA-sequencing data on proximal sections of the same specimens, we identify cell-cell crosstalk associated with lymph node metastasis, demonstrating the power of CELESTA to facilitate identification of clinically relevant interactions.

Ascle-A Python Natural Language Processing Toolkit for Medical Text Generation: Development and Evaluation Study.

BACKGROUND: Medical texts present significant domain-specific challenges, and manually curating these texts is a time-consuming and labor-intensive process. To address this, natural language processing (NLP) algorithms have been developed to automate text processing. In the biomedical field, various toolkits for text processing exist, which have greatly improved the efficiency of handling unstructured text. However, these existing toolkits tend to emphasize different perspectives, and none of them offer generation capabilities, leaving a significant gap in the current offerings. OBJECTIVE: This study aims to describe the development and preliminary evaluation of Ascle. Ascle is tailored for biomedical researchers and clinical staff with an easy-to-use, all-in-one solution that requires minimal programming expertise. For the first time, Ascle provides 4 advanced and challenging generative functions: question-answering, text summarization, text simplification, and machine translation. In addition, Ascle integrates 12 essential NLP functions, along with query and search capabilities for clinical databases. METHODS: We fine-tuned 32 domain-specific language models and evaluated them thoroughly on 27 established benchmarks. In addition, for the question-answering task, we developed a retrieval-augmented generation (RAG) framework for large language models that incorporated a medical knowledge graph with ranking techniques to enhance the reliability of generated answers. Additionally, we conducted a physician validation to assess the quality of generated content beyond automated metrics. RESULTS: The fine-tuned models and RAG framework consistently enhanced text generation tasks. For example, the fine-tuned models improved the machine translation task by 20.27 in terms of BLEU score. In the question-answering task, the RAG framework raised the ROUGE-L score by 18% over the vanilla models. Physician validation of generated answers showed high scores for readability (4.95/5) and relevancy (4.43/5), with a lower score for accuracy (3.90/5) and completeness (3.31/5). CONCLUSIONS: This study introduces the development and evaluation of Ascle, a user-friendly NLP toolkit designed for medical text generation. All code is publicly available through the Ascle GitHub repository. All fine-tuned language models can be accessed through Hugging Face.

Exploring The Impact of Acceptance and Commitment Based Cultural Humility Training on Standardized Patient Interactions: Revisiting the Measurement Process.

Recent findings in health sciences and medical education highlight the importance of training healthcare professionals to interact with their patients in a culturally humble manner (Nadal et al., in Journal of Counseling and Development 92: 57-66, 2014; Pascoe & Smart Richman, in Psychological Bulletin 135: 531, 2009; Sirois & Burg, in Behavior Modification 27: 83-102, 2003; Williams & Mohammed, in Journal of Behavioral Medicine 32: 20-47, 2009). An important piece in the progression of our ability to address training challenges is the assessment of cultural humility. As an extension of previous research (Lombardero et al., in Journal of Clinical Psychology in Medical Settings, 30: 261-273, 2023), this study implemented an evidence-based cultural humility intervention (based on Acceptance and Commitment Training) to improve medical students' interactions with standardized patients (SPs) which was assessed via direct behavioral observation. Specifically, the observational measurement system was focused on culturally humble responses to patients reporting microaggressions to the medical professional. A pre-post comparison of the results demonstrated statistically significant improvements pertaining to participants' culturally humble responses to SPs' reports of microaggressions for one of the measurement scales used (i.e., ARISE), but not the other (i.e., Responsiveness to Racial Challenges Scale). Further analyses, on the bottom quartile of performers, were conducted to assess a possible ceiling effect of the scale that did not demonstrate significant change. These results and implications for future research will be discussed.

Ventricular tachycardia converts to sinus rhythm after administration of propofol.

Ventricular tachycardia (VT) is a major contributor to sudden cardiac death, and pharmacologic treatment options beyond antiarrhythmics are limited. Emerging data suggest sympathetic blocking agents such as propofol are a potential management option for VT refractory to first line antiarrhythmics. Previous literature has described fixed-dose propofol boluses and continuous infusions to convert ventricular arrhythmias; however, to our knowledge, there are no reports of a weight-based dosing strategy for VT. We present the case of a patient with amiodarone-refractory VT who received a 1 mg/kg propofol bolus in preparation for cardioversion and subsequently converted to normal sinus rhythm. The patient stabilized following these interventions, transferred to a tertiary care facility, and was discharged home with an implantable cardioverter defibrillator.

Exosomes in the tumor microenvironment as mediators of cancer therapy resistance.

Exosomes are small extracellular vesicles that contain genetic material, proteins, and lipids. They function as potent signaling molecules between cancer cells and the surrounding cells that comprise the tumor microenvironment (TME). Exosomes derived from both tumor and stromal cells have been implicated in all stages of cancer progression and play an important role in therapy resistance. Moreover, due to their nature as mediators of cell-cell communication, they are integral to TME-dependent therapy resistance. In this review, we discuss current exosome isolation and profiling techniques and their role in TME interactions and therapy resistance. We also explore emerging clinical applications of both exosomes as biomarkers, direct therapeutic targets, and engineered nanocarriers. In order to fully understand the TME, careful interrogation of exosomes and their cargo is critical. This understanding is a promising avenue for the development of effective clinical applications.

Tunable photonic-like modes in graphene-coated nanowires.

In this study, the dispersion equations of a graphene-coated nanowire (GN) are solved. It is found that in this waveguide, besides the surface plasmon polaritons (SPPs), there is another branch of guided modes, called photonic-like modes. The propagation distances of the photonic-like modes can be five orders of magnitude longer than those of the SPPs. Moreover, they can be modulated in the range of 10-4 to 1 m by changing the chemical potential of graphene. In particular, the mode field distributions remain nearly unchanged during the modulation. Based on the analysis performed using COMSOL Multiphysics, we further demonstrated that the propagation losses of the photonic-like modes are dependent on not only the chemical potential of graphene, but also the mode power proportion inside graphene. The photonic-like modes have tremendous potential to be used in optical switches, modulators, and switches in magnetic fields at the nanoscale.

Multiple boluses of alteplase followed by extracorporeal membrane oxygenation for massive pulmonary embolism.

Thrombolytics and extracorporeal membrane oxygenation (ECMO) are potential management options for massive pulmonary embolism (PE). There are early data supporting the use of repeated alteplase 50 mg bolus for massive PE. However, there is sparse literature addressing placement of ECMO catheters after systemic thrombolysis, and there are no reports of initiating ECMO after repeated bolus of alteplase. We present the case of a patient with massive PE who received two boluses of alteplase for recurrent cardiac arrest, followed by initiation of ECMO. The patient stabilized with these interventions, and ultimately had a good outcome with normal neurologic and functional status.

Graphene-coated nanowires with a drop-shaped cross section for 10 nm confinement and 1 mm propagation.

Strong confinement and long-range propagation of electromagnetic energy are longed for when designing efficient miniaturized photonic devices. Here, a graphene-coated nanowire with a drop-shaped cross section is proposed for guiding graphene surface plasmon polaritons to demonstrate an extremely long propagation length (1 mm) with ultra-strong mode confinement (10 nm), which results from the distinctive mode field distribution caused by both the top and bottom arcs of the waveguide. The combination of nanoconcentration and long-range propagation makes the waveguide very useful in nanophotonics, bio-photonics, and highly integrated photonic circuits.

Earlier Alcohol Use Onset Predicts Poorer Neuropsychological Functioning in Young Adults.

BACKGROUND: Neurodevelopment may be shaped by environmental factors such as alcohol intake. Over 20% of U.S. high school students begin drinking before age 14, and those who initiated drinking before age 14 are 4 times more likely to develop psychosocial, psychiatric, and substance use difficulties than those who began drinking after turning 20. Little is known, however, about how the age of alcohol use onset influences brain development. METHODS: This study prospectively examined the effects of alcohol use onset age on neurocognitive functioning in healthy adolescent drinkers (N = 215). Youth were administered a neuropsychological battery before substance use initiation (M = 13.6 years, SD = 0.8) and on average 6.8 years later (M = 20.2 years, SD = 1.5). Hierarchical linear regressions examined if earlier ages of onset for first and regular (i.e., weekly) alcohol use adversely influenced neurocognition, above and beyond baseline neurocognition, substance use severity, and familial and social environment factors. RESULTS: As hypothesized, an earlier age of first drinking onset (AFDO) predicted poorer performance in the domains of psychomotor speed and visual attention (ps<0.05, N = 215) and an earlier age of weekly drinking onset (AWDO) predicted poorer performances on tests of cognitive inhibition and working memory, controlling for baseline neuropsychological performance, drinking duration, and past-year marijuana use (ps<0.05, N = 127). No relationship between AFDO and AWDO was found with verbal learning and memory and visuospatial ability. CONCLUSIONS: This is the first study to assess the association between age of adolescent drinking onset and neurocognitive performance using a comprehensive test battery. This study suggests that early onset of drinking increases risk for alcohol-related neurocognitive vulnerabilities and that initiation of any or weekly alcohol use at younger ages appears to be a risk factor for poorer subsequent neuropsychological functioning. Findings have important implications for public policies related to the legal drinking age and prevention programming. Further studies are needed to replicate these preliminary findings and better understand mediating processes and moderating conditions.

Large-area and highly crystalline MoSe2 for optical modulator.

Transition metal dichalcogenides (TMDs) have been successfully used as broadband optical modulator materials for pulsed fiber laser systems. However, the nonlinear optical absorptions of exfoliated TMDs are strongly limited by their nanoflakes morphology with uncontrollable lateral size and thickness. In this work, we provide an effective method to fully explore the nonlinear optical properties of MoSe2. Large-area and high quality lattice MoSe2 grown by chemical vapor deposition method was adopted as an optical modulator for the first time. The large-area MoSe2 shows excellent nonlinear optical absorption with a large modulation depth of 21.7% and small saturable intensity of 9.4 MW cm-2. After incorporating the MoSe2 optical modulator into fiber laser cavity as a saturable absorber, a highly stable Q-switching operation with single pulse energy of 224 nJ is achieved. The large-area MoSe2 possessing superior nonlinear optical properties compared to exfoliated nanoflakes affords possibility for the larger-area two-dimensional materials family as high performance optical devices.

Thin-wall tubes for coupling terahertz waves to metal wires.

Bare metal wires are among the most promising waveguides for guiding terahertz (THz) surface plasmon polaritons. In this study, a thin-wall tube is proposed for coupling THz waves to a metal wire with ultrahigh efficiency, which results from three high mode matchings for the two waveguides: field distributions, polarization directions, and wave vectors. According to the mode-overlap calculation, the coupling efficiency can be always between 84% and 94% when the frequency of THz waves is in the range of 0.2-3 THz and the metal wire radius is 0.5 mm. The maximum efficiency is as high as 94% at 0.5 THz, which is much higher than that obtained by the previous methods. We further conclude that the optimal coupling efficiency can be obtained when the outer tube radius is equal to the wire radius and simultaneously the real propagation constants of modes in the two waveguides are the same.

Determining the Safety and Tolerability of Rapid Administration of Undiluted Intravenous Levetiracetam in Pediatrics.

Objective: Levetiracetam is widely used in the emergency setting. Safety and tolerability of undiluted levetiracetam is prevalent in adults but is limited in pediatrics. The purpose is to determine the safety and tolerability of rapid administration of undiluted levetiracetam in pediatric patients. Methods: A retrospective, single-center, observational study was conducted in pediatric patients who received undiluted levetiracetam intravenous push. The primary outcome was adverse reactions, extravasation, need for intravenous line replacement, and discontinuation due to adverse reactions. The secondary outcome was turnaround time between ordering and administering first doses. Results: One hundred fourteen patients were included. Injection site reactions occurred in 7 patients. Extravasation occurred in 4 patients. Two patients required intravenous line replacement. There were no adverse events leading to discontinuation of levetiracetam. No difference was seen in the time from order to administration. Conclusion: Rapid administration of undiluted levetiracetam in pediatric patients was safe and well tolerated.

Label-free metabolic optical biomarkers track stem cell fate transition in real time.

Tracking stem cell fate transition is crucial for understanding their development and optimizing biomanufacturing. Destructive single-cell methods provide a pseudotemporal landscape of stem cell differentiation but cannot monitor stem cell fate in real time. We established a metabolic optical metric using label-free fluorescence lifetime imaging microscopy (FLIM), feature extraction and machine learning-assisted analysis, for real-time cell fate tracking. From a library of 205 metabolic optical biomarker (MOB) features, we identified 56 associated with hematopoietic stem cell (HSC) differentiation. These features collectively describe HSC fate transition and detect its bifurcate lineage choice. We further derived a MOB score measuring the "metabolic stemness" of single cells and distinguishing their division patterns. This score reveals a distinct role of asymmetric division in rescuing stem cells with compromised metabolic stemness and a unique mechanism of PI3K inhibition in promoting ex vivo HSC maintenance. MOB profiling is a powerful tool for tracking stem cell fate transition and improving their biomanufacturing from a single-cell perspective.

Medical student attitudes and perceptions of psychedelic-assisted therapies.

Introduction: Although certain psychedelic agents may soon gain federal approval for use in treating specific psychiatric conditions, the utilization of such therapies in clinical practice will depend largely on the attitudes of healthcare providers. Therefore, this study assesses the current attitudes, knowledge, exposure, and acceptance of psychedelics and psychedelic-assisted therapies amongst medical students. Methods: In fall semester of 2022, surveys were emailed to 580 medical students attending medical institutions in the state of Nevada in the United States. Utilizing knowledge and attitude items from previously published studies, the survey collected demographic data and assessed student attitudes with five-point Likert-scale variables. Data was analyzed using summary statistics and Kruskal-Wallis tests for differences in mean survey scores (i.e., attitudes towards psychedelics) based on demographic factors. Results: 132 medical students participated in the survey (22.7% response rate). Medical students demonstrated overall positive attitudes towards psychedelics, lack of knowledge regarding psychedelics, and uncertainty towards neurocognitive risks of psychedelics. Overall, 78.6% of students agreed that psychedelics have therapeutic potential, while 95.2% agreed that psychedelics deserves further research in assessing this potential. Additionally, there was no statistically significant effect of demographic variables, including age, sex, and level of training, on attitudes. Discussion: Although students are overall curious and optimistic about psychedelics, they demonstrate a lack of knowledge regarding recent research efforts. As the field of psychiatry prepares to implement psychedelics and psychedelic-assisted therapies, education and awareness of such agents should be initiated early on in medical clinical training.

Evaluation of phenobarbital for prevention of alcohol withdrawal in trauma patients.

BACKGROUND: Up to 30% of trauma patients experience alcohol withdrawal syndrome (AWS) during their hospital admission, which is associated with worse outcomes. While benzodiazepines and phenobarbital are the mainstay of AWS management, there are limited data on the prevention of AWS. The objective was to evaluate the safety and efficacy of phenobarbital for the prevention of AWS. METHODS: Adult patients admitted to a level 1 trauma center who received at least one dose of phenobarbital for the prevention of AWS between January 2019 and August 2021 were included. Patients were case matched to a control group managed with symptom-triggered therapy based on risk of AWS. Risk factors included sex, age, history of AWS/delirium tremens or withdrawal seizures, selected laboratory values, and screening questionnaires. The primary endpoint was the need for rescue therapy. Secondary endpoints included the time to rescue therapy, intensive care unit (ICU) length of stay (LOS), and hospital LOS. RESULTS: Overall, 110 patients were included with 55 patients in each group. The phenobarbital group had higher baseline Injury Severity Scores ( p = 0.03) and were more likely to be admitted to the ICU (44% vs. 24%; p = 0.03). The phenobarbital group required less rescue therapy (16% vs. 62%; p < 0.001) with a longer time to rescue therapy administration (26 vs. 11 hours; p = 0.01). The phenobarbital group had a longer hospital LOS (216 vs. 87 hours; p = 0.0001) but no difference in ICU LOS ( p = 0.36). There was no incidence of delirium tremens or seizures and no difference in intubation rates ( p = 0.68). There was no incidence of hypotension associated with phenobarbital. CONCLUSION: Patients managed with phenobarbital had a lower need for rescue therapy for AWS with no increased adverse effects. Further studies should evaluate a protocol to prevent alcohol withdrawal in the trauma population. LEVEL OF EVIDENCE: Therapeutic/Care Management; Level III.

The colocatome as a spatial -omic reveals shared microenvironment features between tumour-stroma assembloids and human lung cancer.

Computational frameworks to quantify and compare microenvironment spatial features of in-vitro patient-derived models and clinical specimens are needed. Here, we acquired and analysed multiplexed immunofluorescence images of human lung adenocarcinoma (LUAD) alongside tumour-stroma assembloids constructed with organoids and fibroblasts harvested from the leading edge (Tumour-Adjacent Fibroblasts;TAFs) or core (Tumour Core Fibroblasts;TCFs) of human LUAD. We introduce the concept of the "colocatome" as a spatial -omic dimension to catalogue all proximate and distant colocalisations between malignant and fibroblast subpopulations in both the assembloids and clinical specimens. The colocatome expands upon the colocalisation quotient (CLQ) through a nomalisation strategy that involves permutation analysis and thereby allows comparisons of CLQs under different conditions. Using colocatome analysis, we report that both TAFs and TCFs protected cancer cells from targeted oncogene treatment by uniquely reorganising the tumour-stroma cytoarchitecture, rather than by promoting cellular heterogeneity or selection. Moreover, we show that the assembloids' colocatome recapitulates the tumour-stroma cytoarchitecture defining the tumour microenvironment of LUAD clinical samples and thereby can serve as a functional spatial readout to guide translational discoveries.

Reconstructing codependent cellular cross-talk in lung adenocarcinoma using REMI.

Cellular cross-talk in tissue microenvironments is fundamental to normal and pathological biological processes. Global assessment of cell-cell interactions (CCIs) is not yet technically feasible, but computational efforts to reconstruct these interactions have been proposed. Current computational approaches that identify CCI often make the simplifying assumption that pairwise interactions are independent of one another, which can lead to reduced accuracy. We present REMI (REgularized Microenvironment Interactome), a graph-based algorithm that predicts ligand-receptor (LR) interactions by accounting for LR dependencies on high-dimensional, small-sample size datasets. We apply REMI to reconstruct the human lung adenocarcinoma (LUAD) interactome from a bulk flow-sorted RNA sequencing dataset, then leverage single-cell transcriptomics data to increase the cell type resolution and identify LR prognostic signatures among tumor-stroma-immune subpopulations. We experimentally confirmed colocalization of CTGF:LRP6 among malignant cell subtypes as an interaction predicted to be associated with LUAD progression. Our work presents a computational approach to reconstruct interactomes and identify clinically relevant CCIs.

Results of the phase I CCTG IND.231 trial of CX-5461 in patients with advanced solid tumors enriched for DNA-repair deficiencies.

CX-5461 is a G-quadruplex stabilizer that exhibits synthetic lethality in homologous recombination-deficient models. In this multicentre phase I trial in patients with solid tumors, 40 patients are treated across 10 dose levels (50-650 mg/m2) to determine the recommended phase II dose (primary outcome), and evaluate safety, tolerability, pharmacokinetics (secondary outcomes). Defective homologous recombination is explored as a predictive biomarker of response. CX-5461 is generally well tolerated, with a recommended phase II dose of 475 mg/m2 days 1, 8 and 15 every 4 weeks, and dose limiting phototoxicity. Responses are observed in 14% of patients, primarily in patients with defective homologous recombination. Reversion mutations in PALB2 and BRCA2 are detected on progression following initial response in germline carriers, confirming the underlying synthetic lethal mechanism. In vitro characterization of UV sensitization shows this toxicity is related to the CX-5461 chemotype, independent of G-quadruplex synthetic lethality. These results establish clinical proof-of-concept for this G-quadruplex stabilizer. Clinicaltrials.gov NCT02719977.