Using the High-Entropy Approach to Obtain Multimetal Oxide Nanozymes: Library Synthesis, In Silico Structure-Activity, and Immunoassay Performance.

High-entropy nanomaterials exhibit exceptional mechanical, physical, and chemical properties, finding applications in many industries. Peroxidases are metalloenzymes that accelerate the decomposition of hydrogen peroxide. This study uses the high-entropy approach to generate multimetal oxide-based nanozymes with peroxidase-like activity and explores their application as sensors in ex vivo bioassays. A library of 81 materials was produced using a coprecipitation method for rapid synthesis of up to 100 variants in a single plate. The A and B sites of the magnetite structure, (AA')(BB'B'')2O4, were substituted with up to six different cations (Cu/Fe/Zn/Mg/Mn/Cr). Increasing the compositional complexity improved the catalytic performance; however, substitutions of single elements also caused drastic reductions in the peroxidase-like activity. A generalized linear model was developed describing the relationship between material composition and catalytic activity. Binary interactions between elements that acted synergistically or antagonistically were identified, and a single parameter, the mean interaction effect, was observed to correlate highly with catalytic activity, providing a valuable tool for the design of high-entropy-inspired nanozymes.

Using Smartphone Technology to Track Real-Time Changes in Anxiety/Depression Symptomatology Among Florida Cannabis Users.

Objective: Recent scientific attention has focused on the therapeutic effectiveness of cannabis use on a variety of physical and mental ailments. The present study uses smartphone technology to assess self-reported experiences of Florida cannabis users to understand how cannabis may impact anxiety and depression symptomatology. Method: Several hundred Releaf App™ users from the state of Florida provided anonymous, real-time reports of their symptoms of anxiety and/or depression immediately before and after cannabis use sessions. Linear mixed-effects modeling was used to analyze the data at the symptom and user level. Results: Results showed that for the majority of users, cannabis use was associated with a significant decrease in depression and anxiety symptomatology. While symptom type, doses per session, consumption method, and CBD levels were significant predictors of relief change, their effect sizes were small and should be interpreted with caution. At the user level, those who had positive relief outcomes in anxiety reported more doses and sessions, and those in the depression group reported more sessions. Conclusions: Our results generally support the therapeutic effectiveness of cannabis against depression/anxiety symptomatology. Future work should include standardized statistics and effect size estimates for a better understanding of each variable's practical contribution to this area of study.

Mental Health Stigma in the Workplace and its Association with Possible Actions of Managers to Prevent Sickness Absence of Employees with Mental Health Problems in the Swedish Private Sector: a Video Vignette Study.

PURPOSE: Mental health problems (MHPs) are subjected to workplace stigma and can deteriorate into common mental disorders (CMDs) and sickness absence (SA). Research has shown that personal stigmatizing attitudes limit managers' efforts towards employees with MHPs, but knowledge is lacking regarding stigma in social contexts (contextual stigma) and different types of possible preventive actions. This study investigates personal stigmatizing attitudes and three contextual stigma layers (employee, collegial, organizational) and different types of possible actions to prevent SA of employees with MHPs. METHOD: Survey data of 2769 Swedish managers working in the private sector were analysed. Personal stigmatizing attitudes were measured with the managerial stigma towards employee depression scale and supplemented with four additional items capturing contextual stigma. Managers watched video vignettes and assessed which preventive actions (n = 20) were possible to use in their organization. A sum score was calculated reflecting the 'number of actions'. Principal component analysis revealed three action types: adapt tasks and setting, involve experts, and social support. A score reflecting the 'possibilities to implement actions' was calculated for each type. Multiple linear regression analyses were conducted with the four stigma layers as independent variables for each of the three action variables. RESULTS: Personal stigmatizing attitudes and contextual stigma were significantly associated with both 'number of actions' and 'possibilities for implementing actions' relating to all action types. Patterns of associations with contextual stigma were significant but varied between the different action types. CONCLUSION: This study substantiated the role of personal stigmatizing attitudes and contextual stigma in relation to possible actions of managers to prevent SA of employees with MHPs. The results emphasize the role of contextual stigma. Implications for practice and research are discussed.

A large-scale and PCR-referenced vocal audio dataset for COVID-19.

The UK COVID-19 Vocal Audio Dataset is designed for the training and evaluation of machine learning models that classify SARS-CoV-2 infection status or associated respiratory symptoms using vocal audio. The UK Health Security Agency recruited voluntary participants through the national Test and Trace programme and the REACT-1 survey in England from March 2021 to March 2022, during dominant transmission of the Alpha and Delta SARS-CoV-2 variants and some Omicron variant sublineages. Audio recordings of volitional coughs, exhalations, and speech were collected in the 'Speak up and help beat coronavirus' digital survey alongside demographic, symptom and self-reported respiratory condition data. Digital survey submissions were linked to SARS-CoV-2 test results. The UK COVID-19 Vocal Audio Dataset represents the largest collection of SARS-CoV-2 PCR-referenced audio recordings to date. PCR results were linked to 70,565 of 72,999 participants and 24,105 of 25,706 positive cases. Respiratory symptoms were reported by 45.6% of participants. This dataset has additional potential uses for bioacoustics research, with 11.3% participants self-reporting asthma, and 27.2% with linked influenza PCR test results.

A proof of principle investigation of a novel enzyme formulation on dental calculus deposition: a 4-week randomized human clinical trial.

BACKGROUND: The dissolution of dental calculus, safely and at home, is among the more challenging issues facing the over-the-counter healthcare industry. Pontis Biologics, Inc. has developed novel model of calculus development and structure and has formulated a dentifrice (Tartarase™) using digestive enzymes as active ingredients that is shown to dissolve dental calculus in this Proof of Principle clinical trial. METHODS: This investigation was designed to evaluate the safety and efficacy of a novel enzyme formulation to remove existing calculus deposits in 4 weeks, measured using the Volpe-Manhold Index (V-MI) on lingual surfaces of 6 lower anterior teeth. The test formulation was compared to Crest Cavity Protection, as a control dentifrice. A total of 40 randomized test subjects began the study with 20 assigned to the control dentifrice and 20 assigned to the Tartarase groups (ten each, one brushing with Tartarase twice daily and one brushed with Tartarase and wore a dental tray filled with Tartarase for 30 min then brushed again with Tartarase, once daily). RESULTS: The Crest group experienced a 12% increase in calculus, in contrast to the results of both Tartarase groups that experienced a 40% reduction in calculus in 4 weeks of unsupervised at home use of the Tartarase toothpaste formulation. CONCLUSIONS: This proof of principle study demonstrates that a dentifrice, formulated along the lines of the Tartarase material, is capable of combating calculus accumulation using the same oral hygiene habits that are common worldwide. TRIAL REGISTRATION: This trial was registered retrospectively at clinicaltrials.gov and has the Unique Identification Number: NCT06139835, 14/11/2023.

Medulloblastoma in children with Fanconi anemia: Association with FA-D1/FA-N, SHH type and poor survival independent of treatment strategies.

BACKGROUND: Outcome of children with medulloblastoma (MB) and Fanconi Anemia (FA), an inherited DNA repair deficiency, has not been described systematically. Treatment is complicated by high vulnerability to treatment-associated side effects, yet structured data are lacking. This study aims at giving a comprehensive overview about clinical and molecular characteristics of pediatric FA MB patients. METHODS: Clinical data including detailed information on treatment and toxicities of six previously unreported FA MB patients were supplemented with data of 16 published cases. RESULTS: We identified 22 cases of children with FA and MB with clinical data available. All MBs with subgroup reporting were SHH-activated (n=9), confirmed by methylation profiling in five patients. FA MB patients exclusively belonged to complementation groups FA-D1 (n=16) or FA-N (n=3). Patients were treated with postoperative chemotherapy only (50%) or radiotherapy (RT)±chemotherapy (27%). 23% did not receive adjuvant therapy. Excessive treatment-related toxicities were frequent. Severe hematological toxicity occurred in 91% of patients treated with alkylating chemotherapy, while non-alkylating agents and RT were less toxic. Median overall survival (OS) was 1 year (95%CI 0.3-1.8). 1-year-progression-free-survival (PFS) was 26.3±10.1% and 1-year-OS was 42.1±11.3%. Adjuvant therapy prolonged survival (1y-OS/1y-PFS 0%/0% without adjuvant therapy vs. 53.3±12.9%/33.3±12.2% with adjuvant therapy, p=0.006/p=0.086). CONCLUSIONS: MB in FA patients is strongly associated with SHH activation and FA-D1/FA-N. Despite the dismal prognosis, adjuvant therapy may prolong survival. Non-alkylating chemotherapy and RT are feasible in selected patients with careful monitoring of toxicities and dose adjustments. Curative therapy for FA MB-SHH remains an unmet medical need.

Pre-Transplant Frequencies of FoxP3+CD25+ in CD3+CD8+ T Cells as Potential Predictors for CMV in CMV-Intermediate Risk Kidney Transplant Recipients.

Cytomegalovirus (CMV) infection detrimentally influences graft survival in kidney transplant recipients, with the risk primarily determined by recipient and donor serostatus. However, recipient CD8+ T cells play a crucial role in CMV control. The optimal preventive strategy (prophylaxis vs. pre-emptive treatment), particularly for seropositive (intermediate risk) recipients, remains uncertain. We investigated CD8+ T cell subpopulation dynamics and CMV occurrence (DNAemia ≥ 100 IU/mL) in 65 kidney transplant recipients, collecting peripheral blood mononuclear cells before (T1) and 1 year after transplantation (T2). Comparing the two timepoints, we found an increase in granulocyte, monocyte and CD3+CD8+ T cells numbers, while FoxP3+CD25+, LAG-3+ and PD-1+ frequencies were reduced at T2. CMV DNAemia occurred in 33 recipients (55.8%) during the first year. Intermediate risk patients were disproportionally affected by posttransplant CMV (N = 29/45, 64.4%). Intermediate risk recipients developing CMV after transplantation exhibited lower leukocyte, monocyte, and granulocyte counts and higher FoxP3+CD25+ frequencies in CD3+CD8+ T cells pre-transplantation compared to patients staying CMV negative. Pre-transplant FoxP3+CD25+ in CD3+CD8+ T cells had the best discriminatory potential for CMV infection prediction within the first year after transplantation (AUC: 0.746). The FoxP3+CD25+ CD3+CD8+ T cell subset may aid in selecting intermediate risk kidney transplant recipients for CMV prophylaxis.

Metrology for advanced radiotherapy using particle beams with ultra-high dose rates.

Dosimetry of ultra-high dose rate beams is one of the critical components which is required for safe implementation of FLASH radiotherapy (RT) into clinical practice. In the past years several national and international programmes have emerged with the aim to address some of the needs that are required for translation of this modality to clinics. These involve the establishment of dosimetry standards as well as the validation of protocols and dosimetry procedures. This review provides an overview of recent developments in the field of dosimetry for FLASH RT, with particular focus on primary and secondary standard instruments, and provides a brief outlook on the future work which is required to enable clinical implementation of FLASH RT.

A text-based conversational agent for asthma support: Mixed-methods feasibility study.

Objective: Millions of people in the UK have asthma, yet 70% do not access basic care, leading to the largest number of asthma-related deaths in Europe. Chatbots may extend the reach of asthma support and provide a bridge to traditional healthcare. This study evaluates 'Brisa', a chatbot designed to improve asthma patients' self-assessment and self-management. Methods: We recruited 150 adults with an asthma diagnosis to test our chatbot. Participants were recruited over three waves through social media and a research recruitment platform. Eligible participants had access to 'Brisa' via a WhatsApp or website version for 28 days and completed entry and exit questionnaires to evaluate user experience and asthma control. Weekly symptom tracking, user interaction metrics, satisfaction measures, and qualitative feedback were utilised to evaluate the chatbot's usability and potential effectiveness, focusing on changes in asthma control and self-reported behavioural improvements. Results: 74% of participants engaged with 'Brisa' at least once. High task completion rates were observed: asthma attack risk assessment (86%), voice recording submission (83%) and asthma control tracking (95.5%). Post use, an 8% improvement in asthma control was reported. User satisfaction surveys indicated positive feedback on helpfulness (80%), privacy (87%), trustworthiness (80%) and functionality (84%) but highlighted a need for improved conversational depth and personalisation. Conclusions: The study indicates that chatbots are effective for asthma support, demonstrated by the high usage of features like risk assessment and control tracking, as well as a statistically significant improvement in asthma control. However, lower satisfaction in conversational flexibility highlights rising expectations for chatbot fluency, influenced by advanced models like ChatGPT. Future health-focused chatbots must balance conversational capability with accuracy and safety to maintain engagement and effectiveness.

Pulmonary rehabilitation in follow-up and inpatient rehabilitation for Long COVID: twelve months of follow-up.

BACKGROUND: Individuals with persistent impairments due to Coronavirus disease 2019 (COVID-19) can receive pulmonary rehabilitation in Germany. To date, there is no evidence of the medium- or long-term effects of pulmonary rehabilitation on Long COVID. AIM: This study examined changes in health and occupational outcomes over time and described the therapeutic content of pulmonary rehabilitation and aftercare. This analysis also compared two rehabilitation groups after COVID-19 who had different levels of access to rehabilitation. DESIGN: Longitudinal observational study with multicenter and prospective data collection. SETTING: Pulmonary rehabilitation in four different rehabilitation facilities in Germany. POPULATION: Individuals with a mild course of disease and long-lasting impairments (inpatient rehabilitation, IR) and patients with a severe course after hospitalization (follow-up rehabilitation, FuR). Participants had to be between 18 and 65 years of age. METHODS: Written questionnaires were administered at the beginning and end of rehabilitation, as well as six and twelve months after rehabilitation. Health-related quality of life (HrQoL), fatigue, participation restrictions, COVID-19 symptoms, mental and physical health were assessed, as well as occupational outcomes and questions about rehabilitation and aftercare. RESULTS: IR patients were predominantly female (68.0%) and 52 years of age on average, while 66.1% of Long COVID rehabilitees in FuR were male and three years older. Over the course of rehabilitation, most COVID-19 symptoms decreased with statistical significance. The subjective health scales showed improvements with medium to large effect sizes (ES) over time in IR (P<0.01; ES between 0.55 (cognitive fatigue) and 1.40 (physical fatigue)) and small to large effects in FuR (P<0.01; ES between 0.45 (anxiety) and 1.32 (physical fatigue)). One year after rehabilitation, most effects remained at a moderate level. After twelve months, an increase in neurocognitive symptoms was observed in FuR patients. More than 80% of employed people returned to work one year after rehabilitation, although FuR patients returned to work a median of four weeks later (P<0.01). CONCLUSIONS: The comparative analysis showed that rehabilitees in different forms of rehabilitation attended rehabilitation with different impairments and rehabilitation goals, which are partly considered in treatment and aftercare. CLINICAL REHABILITATION IMPACT: To provide needs-based rehabilitation to different rehabilitation groups with Long COVID, knowledge of their health histories and preferences is necessary.

Immune profiling of age and adjuvant-specific activation of human blood mononuclear cells in vitro.

Vaccination reduces morbidity and mortality due to infections, but efficacy may be limited due to distinct immunogenicity at the extremes of age. This raises the possibility of employing adjuvants to enhance immunogenicity and protection. Early IFNγ production is a hallmark of effective vaccine immunogenicity in adults serving as a biomarker that may predict effective adjuvanticity. We utilized mass cytometry (CyTOF) to dissect the source of adjuvant-induced cytokine production in human blood mononuclear cells (BMCs) from newborns (~39-week-gestation), adults (~18-63 years old) and elders (>65 years of age) after stimulation with pattern recognition receptors agonist (PRRa) adjuvants. Dimensionality reduction analysis of CyTOF data mapped the BMC compartment, elucidated age-specific immune responses and profiled PRR-mediated activation of monocytes and DCs upon adjuvant stimulation. Furthermore, we demonstrated PRRa adjuvants mediated innate IFNγ induction and mapped NK cells as the key source of TLR7/8 agonist (TLR7/8a) specific innate IFNγ responses. Hierarchical clustering analysis revealed age and TLR7/8a-specific accumulation of innate IFNγ producing γδ T cells. Our study demonstrates the application of mass cytometry and cutting-edge computational approaches to characterize immune responses across immunologically distinct age groups and may inform identification of the bespoke adjuvantation systems tailored to enhance immunity in distinct vulnerable populations.

PARP14 and PARP9/DTX3L regulate interferon-induced ADP-ribosylation.

PARP-catalysed ADP-ribosylation (ADPr) is important in regulating various cellular pathways. Until recently, PARP-dependent mono-ADP-ribosylation has been poorly understood due to the lack of sensitive detection methods. Here, we utilised an improved antibody to detect mono-ADP-ribosylation. We visualised endogenous interferon (IFN)-induced ADP-ribosylation and show that PARP14 is a major enzyme responsible for this modification. Fittingly, this signalling is reversed by the macrodomain from SARS-CoV-2 (Mac1), providing a possible mechanism by which Mac1 counteracts the activity of antiviral PARPs. Our data also elucidate a major role of PARP9 and its binding partner, the E3 ubiquitin ligase DTX3L, in regulating PARP14 activity through protein-protein interactions and by the hydrolytic activity of PARP9 macrodomain 1. Finally, we also present the first visualisation of ADPr-dependent ubiquitylation in the IFN response. These approaches should further advance our understanding of IFN-induced ADPr and ubiquitin signalling processes and could shed light on how different pathogens avoid such defence pathways.

Electromembrane extraction of drugs of abuse and prescription drugs from micropulverized hair.

Hair analysis can provide chronological insights into past drug use for months to years after drug administration. In comparison to analyses from other biological matrices, such as blood and urine, sample pretreatment is often tedious and not environmental friendly. In this study, we present a more environmental friendly approach to hair analysis using micropulverized hair and electromembrane extraction for the efficient extraction of 15 drugs of abuse, prescription drugs, and metabolites from hair. The optimized extraction method, involving micropulverization, demonstrated comparable yields to the standard approach of cutting and overnight incubation. A 15-min extraction method using a commercial electromembrane extraction prototype was developed and validated according to forensic guidelines, using only 10 µl of organic solvent per sample. The final method, employing HPLC-MS-MS with a biphenyl column, exhibited good linearity, precision, and sensitivity. An AgreePrep assessment comparing the environmental impact of our method with the standard routine method, involving overnight incubation and conventional liquid-liquid extraction, was conducted. This is the first time micropulverized hair has been subjected to electromembrane extraction.

Molecular and epigenetic ex vivo profiling of testis cancer-associated fibroblasts and their interaction with germ cell tumor cells and macrophages.

Germ cell tumors (GCT) are the most common solid tumors in young men of age 15 - 40. In previous studies, we profiled the interaction of GCT cells with cells of the tumor microenvironment (TM). Earlier studies showed that especially the 3D interaction of fibroblasts (FB) or macrophages with GCT cells influenced the growth behavior and cisplatin response as well as the transcriptome and secretome of the tumor cells, suggesting that the crosstalk of these cells with GCT cells is crucial for tumor progression and therapy outcome. In this study, we shed light on the mechanisms of activation of cancer-associated fibroblasts (CAF) in the GCT setting and their effects on GCT cells lines and the monocyte cell line THP-1. Ex vivo cultures of GCT-derived CAF were established and characterized molecularly and epigenetically by performing DNA methylation arrays, RNA sequencing, and mass spectrometry-based secretome analysis. We demonstrated that the activation state of CAF is influenced by their former prevailing tumor environment in which they have resided. Hereby, we postulated that seminoma (SE) and embryonal carcinoma (EC) activate CAF, while teratoma (TER) play only a minor role in CAF formation. In turn, CAF influence proliferation and the expression of cisplatin sensitivity-related factors in GCT cells lines as well as polarization of in vitro-induced macrophages by the identified effector molecules IGFBP1, LGALS3BP, LYVE1, and PTX3. Our data suggests that the vital interaction of CAF with GCT cells and with macrophages has a huge influence for shaping the extracellular matrix as well as for recruitment of immune cells to the tumor microenvironment. In conclusion, therapeutically interfering with CAF and / or macrophages in addition to the standard therapy might slow-down progression of GCT and re-shaping of the TM to a tumor-promoting environment. Significance: The interaction of CAF with GCT and macrophages considerably influences the microenvironment. Thus, therapeutically interfering with CAF might slow-down progression of GCT and re-shaping of the microenvironment to a tumor-promoting environment.

Synthesis of Structural ADP-Ribose Analogues as Inhibitors for SARS-CoV-2 Macrodomain 1.

Protein adenosine diphosphate (ADP)-ribosylation is crucial for a proper immune response. Accordingly, viruses have evolved ADP-ribosyl hydrolases to remove these modifications, a prominent example being the SARS-CoV-2 NSP3 macrodomain, "Mac1". Consequently, inhibitors are developed by testing large libraries of small molecule candidates, with considerable success. However, a relatively underexplored angle in design pertains to the synthesis of structural substrate mimics. Here, we present the synthesis and biophysical activity of novel adenosine diphosphate ribose (ADPr) analogues as SARS-CoV-2 NSP3 Mac1 inhibitors.

Development of a Flexible Sensor-Integrated Tissue Patch to Monitor Early Organ Rejection Processes Using Impedance Spectroscopy.

Heart failure represents a primary cause of hospitalization and mortality in both developed and developing countries, often necessitating heart transplantation as the only viable recovery path. Despite advances in transplantation medicine, organ rejection remains a significant post-operative challenge, traditionally monitored through invasive endomyocardial biopsies (EMB). This study introduces a rapid prototyping approach to organ rejection monitoring via a sensor-integrated flexible patch, employing electrical impedance spectroscopy (EIS) for the non-invasive, continuous assessment of resistive and capacitive changes indicative of tissue rejection processes. Utilizing titanium-dioxide-coated electrodes for contactless impedance sensing, this method aims to mitigate the limitations associated with EMB, including procedural risks and the psychological burden on patients. The biosensor's design features, including electrode passivation and three-dimensional microelectrode protrusions, facilitate effective monitoring of cardiac rejection by aligning with the heart's curvature and responding to muscle contractions. Evaluation of sensor performance utilized SPICE simulations, scanning electron microscopy, and cyclic voltammetry, alongside experimental validation using chicken heart tissue to simulate healthy and rejected states. The study highlights the potential of EIS in reducing the need for invasive biopsy procedures and offering a promising avenue for early detection and monitoring of organ rejection, with implications for patient care and healthcare resource utilization.

A Review of AIoT-based Human Activity Recognition: From Application to Technique.

This scoping review paper redefines the Artificial Intelligence-based Internet of Things (AIoT) driven Human Activity Recognition (HAR) field by systematically extrapolating from various application domains to deduce potential techniques and algorithms. We distill a general model with adaptive learning and optimization mechanisms by conducting a detailed analysis of human activity types and utilizing contact or non-contact devices. It presents various system integration mathematical paradigms driven by multimodal data fusion, covering predictions of complex behaviors and redefining valuable methods, devices, and systems for HAR. Additionally, this paper establishes benchmarks for behavior recognition across different application requirements, from simple localized actions to group activities. It summarizes open research directions, including data diversity and volume, computational limitations, interoperability, real-time recognition, data security, and privacy concerns. Finally, we aim to serve as a comprehensive and foundational resource for researchers delving into the complex and burgeoning realm of AIoT-enhanced HAR, providing insights and guidance for future innovations and developments.