Corrigendum: Knowledge and thresholds for palliative care and surgery among healthcare providers caring for adults with serious illness.

[This corrects the article DOI: 10.3389/fmed.2024.1351864.].

Spatial Immunophenotyping from Whole-Slide Multiplexed Tissue Imaging Using Convolutional Neural Networks.

The multiplexed immunofluorescence (mIF) platform enables biomarker discovery through the simultaneous detection of multiple markers on a single tissue slide, offering detailed insights into intratumor heterogeneity and the tumor-immune microenvironment at spatially resolved single cell resolution. However, current mIF image analyses are labor-intensive, requiring specialized pathology expertise which limits their scalability and clinical application. To address this challenge, we developed CellGate, a deep-learning (DL) computational pipeline that provides streamlined, end-to-end whole-slide mIF image analysis including nuclei detection, cell segmentation, cell classification, and combined immuno-phenotyping across stacked images. The model was trained on over 750,000 single cell images from 34 melanomas in a retrospective cohort of patients using whole tissue sections stained for CD3, CD8, CD68, CK-SOX10, PD-1, PD-L1, and FOXP3 with manual gating and extensive pathology review. When tested on new whole mIF slides, the model demonstrated high precision-recall AUC. Further validation on whole-slide mIF images of 9 primary melanomas from an independent cohort confirmed that CellGate can reproduce expert pathology analysis with high accuracy. We show that spatial immuno-phenotyping results using CellGate provide deep insights into the immune cell topography and differences in T cell functional states and interactions with tumor cells in patients with distinct histopathology and clinical characteristics. This pipeline offers a fully automated and parallelizable computing process with substantially improved consistency for cell type classification across images, potentially enabling high throughput whole-slide mIF tissue image analysis for large-scale clinical and research applications.

Carbonic anhydrase inhibitors prevent presymptomatic capillary flow disturbances in a model of cerebral amyloidosis.

To enhance early diagnosis and treatment of Alzheimer·s disease (AD), understanding the pathological changes before symptoms arise is crucial. The continuum model of AD suggest that Aß beta (Aß) accumulation precedes symptoms by at least 15 years, with vascular changes detectable around this time. Disturbances in capillary flow dynamics have been linked to reduced oxygen delivery to brain tissue, but evidence in presymptomatic AD remains elusive. We examined capillary flow dynamics in presymptomatic Tg-SwDI mice and the capacity of carbonic anhydrase inhibitors (CAIs) to prevent capillary flow disturbances. Our study revealed capillary flow disturbances associated with alterations in capillary morphology, adhesion molecule expression, and Aß load in cognitively normal 9-10-month-old Tg-SwDI mice. Treated mice showed ameliorated capillary flow disturbances, enhanced oxygen availability, and reduced Aß load. These findings underscore the importance of capillary flow disturbances in presymptomatic AD and highlight CAIs· potential for preserving vascular integrity in early AD.

The Global Evolution and Impact of Systems Biology and Artificial Intelligence in Stem Cell Research and Therapeutics Development: A Scoping Review.

Advanced bioinformatics analysis, such as systems biology (SysBio) and artificial intelligence (AI) approaches, including machine learning (ML) and deep learning (DL), is increasingly present in stem cell (SC) research. An approximate timeline on these developments and their global impact is still lacking. We conducted a scoping review on the contribution of SysBio and AI analysis to SC research and therapy development based on literature published in PubMed between 2000 and 2024. We identified an 8-10-fold increase in research output related to all three search terms between 2000 and 2021, with a 10-fold increase in AI-related production since 2010. Use of SysBio and AI still predominates in preclinical basic research with increasing use in clinically oriented translational medicine since 2010. SysBio- and AI-related research was found all over the globe, with SysBio output led by the United States (US, n=1487), United Kingdom (UK, n=1094), Germany (n=355), The Netherlands (n=339), Russia (n=215), and France (n=149), while for AI-related research the US (n=853) and UK (n=258) take a strong lead, followed by Switzerland (n=69), The Netherlands (n=37), and Germany (n=19). The US and UK are most active in SCs publications related to AI/ML and AI/DL. The prominent use of SysBio in ESC research was recently overtaken by prominent use of AI in iPSC and MSC research. This study reveals the global evolution and growing intersection between AI, SysBio, and SC research over the past two decades, with substantial growth in all three fields and exponential increases in AI-related research in the past decade.

Recommendations for Cardiac Point-of-Care Ultrasound Nomenclature.

Point-of-care ultrasound (POCUS) involves the acquisition, interpretation, and immediate clinical integration of ultrasonographic imaging performed by a treating clinician. The current state of cardiac POCUS terminology is heterogeneous and ambiguous, in part because it evolved through siloed specialty practices. In particular, the medical literature and colloquial medical conversation contain a wide variety of terms that equate to cardiac POCUS. While diverse terminology aided in the development and dissemination of cardiac POCUS throughout multiple specialties, it also contributes to confusion and raises patient safety concerns. This statement is the product of a diverse and inclusive Writing Group from multiple specialties, including medical linguistics, that employed an iterative process to contextualize and standardize a nomenclature for cardiac POCUS. We sought to establish a deliberate vocabulary that is sufficiently unrelated to any specialty, ultrasound equipment, or clinical setting to enhance consistency throughout the academic literature and patient care settings. This statement (1) reviews the evolution of cardiac POCUS-related terms; (2) outlines specific recommendations, distinguishing between intrinsic and practical differences in terminology; (3) addresses the implications of these recommendations for current practice; and (4) discusses the implications for novel technologies and future research.

Ultrasound Probe Pressure Affects Aortic Wall Stiffness: A Patient-Specific Computational Study in Abdominal Aortic Aneurysms.

PURPOSE: Ultrasound imaging is key in the management of patients with an abdominal aortic aneurysm (AAA). It was recently shown that the cyclic diameter variations between diastole and systole, which can be quantified with US imaging, increase significantly with the strength of the applied probe pressure on the patient's abdomen. The goal of this study is to investigate this effect more thoroughly. METHODS: With finite-element modeling, pulsatile blood pressure and probe pressure are simulated in three patient-specific geometries. Two distinct models for the aortic wall were simulated: a nonlinear hyperelastic and a linear elastic model. In addition, varying stiffness was considered for the surrounding tissues. The effect of light, moderate, and firm probe pressure was quantified on the stresses and strains in the aortic wall, and on two in vivo stiffness measures. In addition, the Elasticity Loss Index was proposed to quantify the change in stiffness due to probe pressure. RESULTS: Firm probe pressure decreased the measured aortic stiffness, and material stiffness was affected only when the wall was modeled as nonlinear, suggesting a shift in the stress-strain curve. In addition, stiffer surrounding tissues and a more elongated aneurysm sac decreased the responsiveness to the probe pressure. CONCLUSION: The effect of probe pressure on the AAA wall stiffness was clarified. In particular, the AAA wall nonlinear behavior was found to be of primary importance in determining the probe pressure response. Thus, further work will intend to make use of this novel finding in a clinical context.

A multidisciplinary non-invasive approach to monitor response to intravenous immunoglobulin treatment in neurodegenerative Langerhans cell histiocytosis: a real-world study.

Aims: Early detection and treatment of neurodegenerative Langerhans cell histiocytosis (ND-LCH) have been suggested to prevent neurodegenerative progression. The aim of the study is to validate a standardized multidisciplinary diagnostic work-up to monitor the intravenous immunoglobulins (IVIG) treatment response and the natural course of the disease in untreated patients. Methods: Patients with abnormal somatosensory evoked potentials (SEPs) received monthly 0.5 g/kg IVIG. The diagnostic protocol included structural 3T MRI, neurological examination, brainstem auditory evoked potentials (BAEPs) and SEPs. Results: Twenty-two patients were followed for 5.2 years (median) from the first MRI evidence of ND-LCH. Eleven patients received IVIG for 1.7 years (median). At treatment start neurological examination was abnormal in 10 patients, of whom two had severe clinical impairment and four had abnormal BAEPs. At last follow-up, 1/11 remained stable and 7/11 improved, while worsening of neurological or neurophysiological findings, or both, occurred in 3/11. Risk factors for worsening were a severe clinical or MRI ND-LCH at treatment initiation and prolonged exposure to LCH. Of the 11 untreated patients, none improved and three worsened. Conclusions: Using a standardized diagnostic protocol, we demonstrated that IVIG treatment can lead to clinical stabilization or improvement in all pauci-symptomatic patients with an MRI grading of less than 4.

Changes in REVEAL Lite 2 risk status are associated with long-term outcomes in patients with pulmonary arterial hypertension: a post-hoc analysis of the GRIPHON study.

BACKGROUND: Mortality risk assessment informs clinical management of pulmonary arterial hypertension (PAH). The Registry to Evaluate Early and Long-Term PAH Disease Management (REVEAL) Lite 2 is a simplified risk calculator discriminating 1-year mortality risk. METHODS: This post-hoc analysis of the phase 3 GRIPHON study assessed changes in REVEAL Lite 2 risk score with selexipag versus placebo, and whether changes were prognostic or predictive of time to first morbidity/mortality (M/M) event. RESULTS: REVEAL Lite 2 risk category discriminated M/M risk (landmark concordance indices: 0.68-0.76, selexipag; 0.65-0.70, placebo). Across baseline risk categories, hazard ratios supported a lower risk of M/M events with selexipag versus placebo: low, 0.573 (95% confidence interval [CI] 0.361-0.908; p = 0.0178); intermediate, 0.423 (95% CI 0.274-0.655; p = 0.0001); high, 0.711 (9% CI 0.520-0.972; p = 0.0326). Odds ratios for risk improvement were 2.0 (95% CI 1.50-2.65), 1.8 (95% CI 1.38-2.43), and 2.0 (95% CI 1.43-2.72) for selexipag versus placebo at 16, 26, and 52 weeks, respectively (all p < 0.001). REVEAL Lite 2 risk improvement at Week 16 explained 19.1% of the treatment effect in all patients and 47.0% in patients with REVEAL Lite 2 baseline risk score of ≥ 7. CONCLUSIONS: REVEAL Lite 2 can monitor PAH M/M risk and facilitate treatment optimization. Baseline REVEAL Lite 2 risk score was prognostic of M/M risk in patients with PAH and mediates treatment effect up to 47% for those at higher risk. Lower M/M risk with selexipag versus placebo occurred irrespective of baseline risk category. (ClinicalTrials.gov identifier: NCT01106014).

Social Support, Coping Strategies, Depression, Anxiety, and Cognitive Function Among People With Type 2 Diabetes Mellitus: A Path Analysis.

OBJECTIVE: To explore the linear associations between social support, coping strategies, depression, anxiety, and cognitive function among people with type 2 diabetes mellitus (T2DM) using a path-analytic method. METHODS: This cross-sectional study enrolled 496 individuals hospitalized due to T2DM. Well-trained investigators conducted face-to-face interviews with the participants using the Social Support Rating Scale, the Chinese version of Medical Coping Modes Questionnaire, the Hospital Anxiety and Depression scale, and the Mini Mental State Examination to measure social support (including objective support, subjective support, and support utilization), coping strategies (including confrontation, avoidance, and acceptance-resignation), depression/anxiety, and cognitive function, respectively. A path analysis was used to elucidate the linear associations between social support, coping strategies, depression, anxiety, and cognitive function. RESULTS: In the final path model with satisfactory model fit, objective support was found to be associated with cognitive function not only directly but also indirectly through confrontation coping and depression, and acceptance-resignation coping and depression/anxiety. Further, subjective support was found to be associated with cognitive function indirectly through depression/anxiety, as well as serially through acceptance-resignation coping and depression/anxiety. Support utilization was found to be associated with cognitive function indirectly through confrontation coping and depression, as well as through acceptance-resignation coping and depression/anxiety. CONCLUSION: Social support, coping strategies, depression, and anxiety were associated with cognitive function among people with T2DM, and these associations were best explained by a serial mediation model from social support, coping strategies, and depression and anxiety to cognitive function.

Graphitic nanoflakes modulate the structure and binding of human amylin.

Human amylin is an inherently disordered protein whose ability to form amyloid fibrils is linked to the onset of type II diabetes. Graphitic nanomaterials have potential in managing amyloid diseases as they can disrupt protein aggregation processes in biological settings, but optimising these materials to prevent fibrillation is challenging. Here, we employ bias-exchange molecular dynamics simulations to systematically study the structure and adsorption preferences of amylin on graphitic nanoflakes that vary in their physical dimensions and surface functionalisation. Our findings reveal that nanoflake size and surface oxidation both influence the structure and adsorption preferences of amylin. The purely hydrophobic substrate of pristine graphene (PG) nanoflakes encourages non-specific protein adsorption, leading to unrestricted lateral mobility once amylin adheres to the surface. Particularly on larger PG nanoflakes, this induces structural changes in amylin that may promote fibril formation, such as the loss of native helical content and an increase in ß-sheet character. In contrast, oxidised graphene nanoflakes form hydrogen bonds between surface oxygen sites and amylin, and as such restricting protein mobility. Reduced graphene oxide (rGO) flakes, featuring lower amounts of surface oxidation, are amphiphilic and exhibit substantial regions of bare carbon which promote protein binding and reduced conformational flexibility, leading to conservation of the native structure of amylin. In comparison, graphene oxide (GO) nanoflakes, which are predominantly hydrophilic and have a high degree of surface oxidation, facilitate considerable protein structural variability, resulting in substantial contact area between the protein and GO, and subsequent protein unfolding. Our results indicate that tailoring the size, oxygen concentration and surface patterning of graphitic nanoflakes can lead to specific and robust protein binding, ultimately influencing the likelihood of fibril formation. These atomistic insights provide key design considerations for the development of graphitic nanoflakes that can modulate protein aggregation by sequestering protein monomers in the biological environment and inhibit conformational changes linked to amyloid fibril formation.

Suicide decision-making: Differences in proximal considerations between individuals who aborted and attempted suicide.

INTRODUCTION: The transition from suicidal thoughts to behaviors often involves considering the consequences of suicide as part of the decision-making process. This study explored the relationship between this consideration process and the decision to either abort or carry out a suicide attempt. METHODS: Among inpatients with a suicide-related event in the past 2 weeks (suicide attempt n = 30 or aborted attempt n = 16), we assessed the degree to which they considered six domains of consequences, the impact of these considerations on their inclination to attempt suicide, and the duration of their decision-making. RESULTS: All the participants who aborted and 87% of those who attempted considered consequences of suicide. Participants who aborted took longer to progress through decision-making stages and considered more suicide-hindering factors, especially interpersonal ones, though these differences were no longer significant after correction. Group status moderated the relationship between the balance of suicide-facilitating and suicide-hindering considerations and decision-making duration. Considering the consequences of suicide more favorably was related to a shorter ideation-to-action period before a suicide attempt and a longer ideation period before aborting an attempt. CONCLUSION: This study highlights the complexity of suicide decision-making and its role in better understanding the progression from ideation to action.

Transcriptomic Analysis Reveals the Heterogeneous Role of Conducting Films Upon Electrical Stimulation.

Central nervous system (CNS) injuries and neurodegenerative diseases have markedly poor prognoses and can result in permanent dysfunction due to the general inability of CNS neurons to regenerate. Differentiation of transplanted stem cells has emerged as a therapeutic avenue to regenerate tissue architecture in damaged areas. Electrical stimulation is a promising approach for directing the differentiation outcomes and pattern of outgrowth of transplanted stem cells, however traditional inorganic bio-electrodes can induce adverse effects such as inflammation. This study demonstrates the implementation of two organic thin films, a polymer/reduced graphene oxide nanocomposite (P(rGO)) and PEDOT:PSS, that have favorable properties for implementation as conductive materials for electrical stimulation, as well as an inorganic indium tin oxide (ITO) conductive film. Transcriptomic analysis reveals that electrical stimulation improves neuronal differentiation of SH-SY5Y cells on all three films, with the greatest effect for P(rGO). Unique material- and electrical stimuli-mediated effects are observed, associated with differentiation, cell-substrate adhesion, and translation. The work demonstrates that P(rGO) and PEDOT:PSS are highly promising organic materials for the development of biocompatible, conductive scaffolds that will enhance electrically-aided stem cell therapeutics for CNS injuries and neurodegenerative diseases.

Clinical and echocardiographic predictors of outcome in liver transplant patients.

Liver transplant (LT) candidates undergo transthoracic echocardiography (TTE) before surgery to assess cardiac function and evaluate the echocardiographic probability of pulmonary hypertension (PHT). The improvement of pulmonary artery systolic pressure (PAPS) after transplant is associated with higher survival rates in patients with mild or moderate PHT. Although studies analyze the outcomes of LT in patients treated for PHT, the prognostic value of PAPS in patients without PHT in the follow-up is unknown. The aim of our study is to evaluate pre- and post-LT cardiac function, right ventricular function, pulmonary artery pressure, and their association with long-term mortality. 102 patients who underwent LT between 2011 and 2018 were compared for echocardiographic and hemodynamic parameters pre- and post-LT. After LT, systolic blood pressure, heart rate (HR), and PASP significantly increased, while tricuspid annular plane systolic excursion/PAPS decreased. Moreover, the higher difference in HR and PASP between pre- and post-LT was highlighted in those patients who died during the follow-up period after LT. Among all the parameters tested, in the multi-variable Cox regression for mortality, left ventricular ejection fraction and PASP difference were predictors of mortality. This study highlights the importance of TTE in LT screening as a tool to stratify patients at higher risk of death due to advanced cirrhotic cardiomyopathy and the importance of the change of echocardiographic parameters, in particular right and left ventricular hemodynamics, during the follow-up period. These parameters could be used to guide a more aggressive therapy.

Shaping psychotherapy trainees' potential: insights from training program directors.

The role of the therapist has received growing attention in psychotherapy research, suggesting that training effectiveness may also depend on the person of the trainees, with relevant implications in terms of candidate selection or tailoring training to the person. In the present study, we focused on how and how much psychotherapy training can be effective in fostering trainees' characteristics associated with successful therapists and contrast trainees' characteristics that could represent limitations as therapists. The aim was to explore training program directors' perspectives on individual trainees' limitations and strengths, and on the effectiveness of training in shaping successful therapists. To this aim, we interviewed 14 training program directors with different psychotherapy approaches. Audio recordings of these semi-structured interviews were transcribed verbatim and analysed using NVivo software. According to our findings, transversally to different psychotherapy approaches, trainees' self-awareness and the ability to embrace uncertainty were recognized as the main characteristics of good trainees. Four training elements were fre- quently mentioned by directors as effective in shaping trainees' development: experiential learning, the use of paradigm that pri- oritizes self-awareness (instead of technique-focused approaches), the centrality of supervision and inter-vision, the trainer-trainee relationship. These insights highlight the importance of trainees' self-awareness and the ability to embrace uncertainty as potential personal variables that may influence the effectiveness of future psychotherapists and suggest giving attention to experiential learning and training relationships as crucial elements of psychotherapists' development during the training.

Understanding, assessing and treating immune, endothelial and haemostasis dysfunctions in bacterial sepsis.

The interplay between the immune system, coagulation, and endothelium is critical in regulating the host response to infection. However, in sepsis and other critical illnesses, a dysregulated immune response can lead to excessive alterations in these mechanisms, resulting in coagulopathy, endothelial dysfunction, and multi-organ dysfunction. This review aims to provide a comprehensive analysis of the pathophysiological mechanisms that govern the complex interplay between immune dysfunction, endothelial dysfunction, and coagulation in sepsis. It emphasises clinical significance, evaluation methods, and potential therapeutic interventions. Understanding these mechanisms is essential for developing effective treatments that can modulate the immune response, mitigate thrombosis, restore endothelial function, and ultimately improve patient survival.

Cardiologists' perspective on termination of pacemaker therapy-an anonymous survey among cardiologists in Germany.

BACKGROUND: The patient's right to refuse pacemaker therapy is mentioned in the relevant European consensus statement but additional information is only available on deactivation of implantable cardioverter deactivator and not on other cardiac implantable electronic devices such as pacemakers. Therefore, we were interested in opinions, concerns and attitudes of cardiologists, who are the primary contact persons for such requests, since the number of patients asking for withdrawal of pacemaker therapy is likely to increase leaving cardiologists and healthcare professionals with a difficult medical but also ethical problem. METHODS: An anonymous questionnaire was sent to all German cardiology departments (N = 288). RESULTS: 48% of cardiology departments responded by sending back 247 completed questionnaires. Most participating cardiologists were experienced when considering the duration of their professional activity. Almost all of the respondents regularly perform check-ups of pacemakers. The majority of cardiologists answering our questionnaire were prepared to deactivate a pacemaker upon patients' request, and have done so. In pacemaker dependency, however, the willingness to withdraw decreases, even if death is imminent, for fear of causing distressing symptoms, sense of being responsible for patients possible immediate death, or fear of legal consequences. CONCLUSIONS: The survey could clearly show that uncertainties remain among cardiologists dealing with a patient's wish for withdrawal, especially in cases of pacemaker dependency. We suggest that official statements of cardiologic societies in Europe are issued to clarify ethical, legal and practical aspects of pacemaker withdrawal. TRIAL REGISTRATION: Registered in the German Clinical Trials Register (DRKS00026168) on 30.08.2021.

Hypoxia evokes a sequence of raphe-pontomedullary network operations for inspiratory drive amplification and gasping.

Hypoxia can trigger a sequence of breathing-related behaviors, from augmentation to apneusis to apnea and gasping. Gasping is an autoresuscitative behavior that, via large tidal volumes and altered intrathoracic pressure, can enhance coronary perfusion, carotid blood flow, and sympathetic activity, and thereby coordinate cardiac and respiratory functions. We tested the hypotheses that hypoxia-evoked gasps are amplified through a disinhibitory microcircuit within the inspiratory neuron chain, and that this drive is distributed via an efference copy mechanism. This generates coordinated gasp-like discharges concurrently in other circuits of the raphe-pontomedullary respiratory network. Data were obtained from 6 decerebrate, vagotomized, neuromuscularly-blocked, and artificially ventilated adult cats. Arterial blood pressure, phrenic nerve activity, end-tidal CO2, and other parameters were monitored. Hypoxia was produced by ventilation with a gas mixture of 5% O2 in nitrogen. Neuron spike trains were recorded at multiple pontomedullary sites simultaneously and evaluated for firing rate modulations and short-time scale correlations indicative of functional connectivity. Experimental perturbations evoked reconfiguration of raphe-pontomedullary circuits during initial augmentation, apneusis and augmented bursts, apnea, and gasping. Functional connectivity, altered firing rates, efference copy of gasp drive, and coordinated incremental blood pressure increases support a distributed brain stem network model for amplification and broadcasting of inspiratory drive during autoresuscitative gasping. Gasping begins with a reduction in inhibition by expiratory neurons and an initial loss of inspiratory drive during hypoxic apnea, and culminates in autoresuscitative efforts.

Temporal trends of suicidality among hospitalised adolescents during COVID-19 pandemic: A Bayesian framework.

OBJECTIVE: Literature on temporal patterns of suicidality among youths during the COVID-19 pandemic is growing. The present work proposes a Bayesian approach to assess temporal patterns of suicide-related behaviours among inpatient adolescents during the COVID-19 pandemic. METHODOLOGY: Data referred to the first hospital discharge record with ICD9-CM codes related to suicide-related behaviour and/or suicidal ideation among adolescents aged 13-19 between 1 January 2017 and 31 March 2021 were collected in the Piedmont region, Italy (n = 334; median age: 15 years, IQR: 14-16; 80% girls). A Poisson Bayesian regression model performed on pre-COVID-19 data (2017-2020), adjusted by seasonality and stratified by sex, was adopted to provide the probability that the predicted counts exceed the observed ones in each pandemic year quarter. RESULTS: A declining trend of suicidality was observed in April-June 2020 among both sex groups. Among females, an increasing pattern of suicidality was registered in early 2021 (January-March) compared to the pre-pandemic period. CONCLUSION: The present findings contributed to a growing literature on the COVID-19 pandemic's impact on adolescents' suicide-related behaviours from a gender perspective and encouraged wider adoption of Bayesian approaches as valuable tools to explore rare events and deeply enlighten open public health issues.