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Abstract Background Digital technologies allow users to engage in health-related behaviors associated with positive outcomes. We aimed to identify classes of US adults with distinct digital technologies access and health use patterns and characterize class composition. Data came from Health Information National Trends Survey Wave 5 Cycles 1–4, a nationally representative cross-sectional survey of US adults (N = 13,993). We used latent class analysis to identify digital technologies access and health use patterns based on 32 ternary variables of behaviors and access to requisite technologies and platforms, including the internet, internet-enabled devices, health monitors, and electronic health records (EHRs). We ran a multinomial logistic regression to identify sociodemographic and health correlates of class membership (n = 10,734). Results Ten classes captured patterns of digital technology access and health use among US adults. This included a digitally isolated, a mobile-dependent, and a super user class, which made up 8.9%, 7.8%, and 13.6% of US adults, respectively, and captured access patterns from only basic cellphones and health monitors to near complete access to web-, mobile-, and EHR-based platforms. Half of US adults belonged to classes that lacked access to EHRs and relied on alternative web-based tools typical of patient portals. The proportion of class members who used digital technologies for health purposes varied from small to large. Older and less educated adults had lower odds of belonging to classes characterized by access or engagement in health behaviors. Hispanic and Asian adults had higher odds of belonging to the mobile-dependent class. Individuals without a regular healthcare provider and those who had not visited a provider in the past year were more likely to belong to classes with limited digital technologies access or health use. Discussion Only one third of US adults belonged to classes that had near complete access to digital technologies and whose members engaged in almost all health behaviors examined. Sex, age, and education were associated with membership in classes that lacked access to 1 + digital technologies or exhibited none to limited health uses of such technologies. Results can guide efforts to improve access and health use of digital technologies to maximize associated health benefits and minimize disparities.

Abstract Ventilatory thresholds (VTs) are key physiological parameters used to evaluate physical performance and determine aerobic and anaerobic transitions during exercise. Current assessment of these parameters requires ergospirometry, limiting evaluation to laboratory or clinical settings. In this work, we introduce a wearable respiratory system that continuously tracks breathing during exercise and estimates VTs during ramp tests. We validate the respiratory rate and VTs predictions in 17 healthy adults using ergospirometry analysis. In addition, we use the wearable system to evaluate VTs in 107 recreational athletes during ramp tests outside the laboratory and show that the mean population values agree with physiological variables traditionally used to exercise prescription. We envision that respiratory wearables can be useful in determining aerobic and anaerobic parameters with promising applications in health telemonitoring and human performance.

ABSTRACT In the era of antimicrobial resistance, phage-antibiotic combinations offer a promising therapeutic option, yet research on their synergy and antagonism is limited. This study aims to assess these interactions, focusing on protein synthesis inhibitors and cell envelope-active agents against multidrug-resistant bacterial strains. We evaluated synergistic and antagonistic interactions in multidrug-resistant Staphylococcus aureus, Enterococcus faecium, and Pseudomonas aeruginosa strains. Phages were combined with protein synthesis inhibitors [linezolid (LZD), minocycline (MIN), gentamicin (GEN), and azithromycin (AZM)] or cell envelope-active agents [daptomycin (DAP), ceftaroline (CPT), and cefepime (FEP)]. Modified checkerboard minimum inhibitory concentration assays and 24-h time-kill analyses were conducted, alongside one-step growth curves to analyze phage growth kinetics. Statistical comparisons used one-way analysis of variance (ANOVA) and the Tukey test (P < 0.05). In the checkerboard and 24-h time-kill analyses (TKA) of S. aureus and E. faecium, phage-LZD and phage-MIN combinations were antagonistic (FIC > 4) while phage-DAP and phage-CPT were synergistic (FIC 0.5) (ANOVA range of mean differences 0.52–2.59 log10 CFU/mL; P < 0.001). For P. aeruginosa, phage-AZM was antagonistic (FIC > 4), phage-GEN was additive (FIC = 1), and phage-FEP was synergistic (ANOVA range of mean differences 1.04–1.95 log10 CFU/mL; P < 0.001). Phage growth kinetics were altered in the presence of LZD and MIN against S. aureus and in the presence of LZD against a single E. faecium strain (HOU503). Our findings indicate that select protein synthesis inhibitors may induce phage-antibiotic antagonism. However, this antagonism may not solely stem from changes in phage growth kinetics, warranting further investigation into the complex interplay among strains, phage attributes, and antibiotic mechanisms affecting bacterial inhibition.IMPORTANCEIn the face of escalating antimicrobial resistance, combining phages with antibiotics offers a promising avenue for treating infections unresponsive to traditional antibiotics. However, while studies have explored synergistic interactions, less attention has been given to potential antagonism and its impact on phage growth kinetics. This research evaluates the interplay between phages and antibiotics, revealing both synergistic and antagonistic patterns across various bacterial strains and shedding light on the complex dynamics that influence treatment efficacy. Understanding these interactions is crucial for optimizing combination therapies and advancing phage therapy as a viable solution for combating antimicrobial resistance.

Abstract Mesenchymal stromal cells (MSCs) are promising regenerative therapeutics that primarily exert their effects through secreted extracellular vesicles (EVs). These EVs – being small and non‐living – are easier to handle and possess advantages over cellular products. Consequently, the therapeutic potential of MSC‐EVs is increasingly investigated. However, due to variations in MSC‐EV manufacturing strategies, MSC‐EV products should be considered as highly diverse. Moreover, the diverse array of EV characterisation technologies used for MSC‐EV characterisation further complicates reliable interlaboratory comparisons of published data. Consequently, this study aimed to establish a common method that can easily be used by various MSC‐EV researchers to characterise MSC‐EV preparations to facilitate interlaboratory comparisons. To this end, we conducted a comprehensive inter‐laboratory assessment using a novel multiplex bead‐based EV flow cytometry assay panel. This assessment involved 11 different MSC‐EV products from five laboratories with varying MSC sources, culture conditions, and EV preparation methods. Through this assay panel covering a range of mostly MSC‐related markers, we identified a set of cell surface markers consistently positive (CD44, CD73 and CD105) or negative (CD11b, CD45 and CD197) on EVs of all explored MSC‐EV preparations. Hierarchical clustering analysis revealed distinct surface marker profiles associated with specific preparation processes and laboratory conditions. We propose CD73, CD105 and CD44 as robust positive markers for minimally identifying MSC‐derived EVs and CD11b, CD14, CD19, CD45 and CD79 as reliable negative markers. Additionally, we highlight the influence of culture medium components, particularly human platelet lysate, on EV surface marker profiles, underscoring the influence of culture conditions on resulting EV products. This standardisable approach for MSC‐EV surface marker profiling offers a tool for routine characterisation of manufactured EV products in pre‐clinical and clinical research, enhances the quality control of MSC‐EV preparations, and hopefully paves the way for higher consistency and reproducibility in the emerging therapeutic MSC‐EV field.

Abstract Prey metabarcoding has become a popular tool in molecular ecology for resolving trophic interactions at high resolution, from various sample types and animals. To date, most predator–prey studies of small‐sized animals (

The aim of the article is to study the role and significance of brand management as the basis for the development of State values in the market. In modern marketing and in the conditions of competition in the market, topical issues are the development of the brand as an important marketing tool that plays a leading role for companies and the State in general in the Ukrainian and international markets. The visual appeal of products and services makes a primary impression on consumers, and therefore the priorities of the brand development strategy are obvious. Understanding the terms «brand» and «brand management» allows you to effectively apply these marketing tools in practice. In the new global digital economy, brands, in cooperation with management, make up a large part of the value of the company itself and a significant source of income. The brand is a challenge of the present and the future and is a mechanism for managing several areas at once: marketing, social, and economic. In today’s international environment, the interaction of brand and management is developing intensively, adapting to rapid changes in the world market and the introduction of information and communication technologies. For the effective functioning of marketing and development of Ukrainian companies that are focused on the market and influence the economy, it is necessary to consider in detail the issue of the functioning of the brand as an innovative tool both at the theoretical and practical levels. In particular, important components are the direct implementation of this mechanism, the list and content of the main stages of application, signs and types, the use of the dynamics of growth of indicators in both the domestic and the international markets. This study presents the features of the use of an innovative brand tool, which are the needs of modern companies, their specifics, overall impact, and also makes forecasts for the future. The use of brand management has a positive effect on the functioning and presentation of Ukraine in the market.

Abstract Extracellular vesicles (EVs) are cell derived membranous nanoparticles. EVs are important mediators of cell–cell communication via the transfer of bioactive content and as such they are being investigated for disease diagnostics as biomarkers and for potential therapeutic cargo delivery to recipient cells. However, existing methods for isolating EVs from biological samples suffer from challenges related to co‐isolation of unwanted materials such as proteins, nucleic acids, and lipoproteins. In the pursuit of improved EV isolation techniques, we introduce multimodal flowthrough chromatography (MFC) as a scalable alternative to size exclusion chromatography (SEC). The use of MFC offers significant advantages for purifying EVs, resulting in enhanced yields and increased purity with respect to protein and nucleic acid co‐isolates from conditioned 3D cell culture media. Compared to SEC, significantly higher EV yields with similar purity and preserved functionality were also obtained with MFC in 2D cell cultures. Additionally, MFC yielded EVs from serum with comparable purity to SEC and similar apolipoprotein B content. Overall, MFC presents an advancement in EV purification yielding EVs with high recovery, purity, and functionality, and offers an accessible improvement to researchers currently employing SEC.

Despite efforts to promote inclusivity in schools, ethnic bullying is a pervasive issue. Recent studies have highlighted the potential of empathy and cultural self efficacy within cross-cultural friendships to mitigate ethnic bullying. Violence prevention programs in the United States rely on didactic approaches and few include embodied approaches. Our goal was to examine the impact of dance/movement therapy (DMT) on empathy, peer relationships, and cultural self-efficacy among racially/ethnically diverse groups of middle school students using a mixed methods research design. Quantitative data did not demonstrate significant change in empathy, peer relationships, and cultural self-efficacy. Qualitative data described an increase in participants’ emotional intelligence including empathy, cultural self-efficacy, and positive cross-cultural relationships. Additional research is needed to determine the definitive impact of DMT on these outcomes.

Abstract Cardiac progenitor cell (CPC)‐derived small extracellular vesicles (sEVs) exhibit great potential to stimulate cardiac repair. However, the multifaceted nature of sEV heterogeneity presents a challenge in understanding the distinct mechanisms underlying their regenerative abilities. Here, a dual‐step multimodal flowthrough and size‐exclusion chromatography method was applied to isolate and separate CPC‐derived sEV subpopulations to study the functional differences related to cardiac repair responses. Three distinct sEV subpopulations were identified with unique protein profiles. Functional cell assays for cardiac repair‐related processes demonstrated that the middle‐sized and smallest‐sized sEV subpopulations exhibited the highest pro‐angiogenic and anti‐fibrotic activities. Proteasome activity was uniquely seen in the smallest‐sized subpopulation. The largest‐sized subpopulation showed no effect in any of the functional assays. This research uncovers the existence of sEV subpopulations, each characterized by a distinct composition and biological function. Enhancing our understanding of sEV heterogeneity will provide valuable insights into sEV mechanisms of action, ultimately accelerating the translation of sEV therapeutics.

Abstract The loss‐of‐function of the proprotein convertase subtilisin–kexin type 9 (Pcsk9) gene has been associated with significant reductions in plasma serum low‐density lipoprotein cholesterol (LDL‐C) levels. Both CRISPR/Cas9 and CRISPR‐based editor‐mediated Pcsk9 inactivation have successfully lowered plasma LDL‐C and PCSK9 levels in preclinical models. Despite the promising preclinical results, these studies did not report how vehicle‐mediated CRISPR delivery inactivating Pcsk9 affected low‐density lipoprotein receptor recycling in vitro or ex vivo. Extracellular vesicles (EVs) have shown promise as a biocompatible delivery vehicle, and CRISPR/Cas9 ribonucleoprotein (RNP) has been demonstrated to mediate safe genome editing. Therefore, we investigated EV‐mediated RNP targeting of the Pcsk9 gene ex vivo in primary mouse hepatocytes. We engineered EVs with the rapamycin‐interacting heterodimer FK506‐binding protein (FKBP12) to contain its binding partner, the T82L mutant FKBP12‐rapamycin binding (FRB) domain, fused to the Cas9 protein. By integrating the vesicular stomatitis virus glycoprotein on the EV membrane, the engineered Cas9 EVs were used for intracellular CRISPR/Cas9 RNP delivery, achieving genome editing with an efficacy of ±28.1% in Cas9 stoplight reporter cells. Administration of Cas9 EVs in mouse hepatocytes successfully inactivated the Pcsk9 gene, leading to a reduction in Pcsk9 mRNA and increased uptake of the low‐density lipoprotein receptor and LDL‐C. These readouts can be used in future experiments to assess the efficacy of vehicle‐mediated delivery of genome editing technologies targeting Pcsk9. The ex vivo data could be a step towards reducing animal testing and serve as a precursor to future in vivo studies for EV‐mediated CRISPR/Cas9 RNP delivery targeting Pcsk9.

The HeartMate 3 Risk Score (HM3RS) is a tool derived and validated from the MOMENTUM 3 clinical trial population to predict mortality after left ventricular assist device (LVAD) implantation. However, the UNOS adult heart allocation policy was revised in October 2018, resulting in a drastic decrease in bridge-to-transplant LVAD use. In this study, we assessed the predictive accuracy of HM3RS at a single institution, both before and after the 2018 UNOS allocation policy change. We retrospectively analyzed all adult patients who received a HeartMate 3 (HM3) at Washington University School of Medicine in St. Louis between 2015 - 2021. We calculated the HM3RS for each patient, categorized as low (<1.58), medium (1.58-2.48), or high (>2.48). The primary outcome was mortality at 1 and 2 years, stratified before and after the date of the UNOS allocation policy change (10/1/2018). A total of 178 HM3 recipients were included, 61 of whom received a HM3 before 10/1/2018. Patient demographics were similar, although LVAD recipients were more likely to be INTERMACS 1 or 2 post-allocation system change compared to pre-allocation system change (78% vs. 47%). HM3RS accurately predicted 1- and 2-year mortality pre-10/2018 with AUC 0.76 and 0.71, respectively, comparable to the original study. However, post-allocation system change, HM3RS performed much worse, with AUC 0.56 at 1 year and 0.40 at 2 years, despite similar patient outcomes (Figure 1). HM3RS accurately predicted mortality in a real-world population before the 2018 UNOS allocation change although it had mediocre performance after. This score may need recalibration to better assess risk for current and future patients undergoing HM3 implantation. [ABSTRACT FROM AUTHOR]

Mesenchymal stromal cells (MSCs) are promising regenerative therapeutics that primarily exert their effects through secreted extracellular vesicles (EVs). These EVs - being small and non-living - are easier to handle and possess advantages over cellular products. Consequently, the therapeutic potential of MSC-EVs is increasingly investigated. However, due to variations in MSC-EV manufacturing strategies, MSC-EV products should be considered as highly diverse. Moreover, the diverse array of EV characterisation technologies used for MSC-EV characterisation further complicates reliable interlaboratory comparisons of published data. Consequently, this study aimed to establish a common method that can easily be used by various MSC-EV researchers to characterise MSC-EV preparations to facilitate interlaboratory comparisons. To this end, we conducted a comprehensive inter-laboratory assessment using a novel multiplex bead-based EV flow cytometry assay panel. This assessment involved 11 different MSC-EV products from five laboratories with varying MSC sources, culture conditions, and EV preparation methods. Through this assay panel covering a range of mostly MSC-related markers, we identified a set of cell surface markers consistently positive (CD44, CD73 and CD105) or negative (CD11b, CD45 and CD197) on EVs of all explored MSC-EV preparations. Hierarchical clustering analysis revealed distinct surface marker profiles associated with specific preparation processes and laboratory conditions. We propose CD73, CD105 and CD44 as robust positive markers for minimally identifying MSC-derived EVs and CD11b, CD14, CD19, CD45 and CD79 as reliable negative markers. Additionally, we highlight the influence of culture medium components, particularly human platelet lysate, on EV surface marker profiles, underscoring the influence of culture conditions on resulting EV products. This standardisable approach for MSC-EV surface marker profiling offers a tool for routine characterisation of manufactured EV products in pre-clinical and clinical research, enhances the quality control of MSC-EV preparations, and hopefully paves the way for higher consistency and reproducibility in the emerging therapeutic MSC-EV field., (© 2024 The Author(s). Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)

Extracellular vesicles (EVs) are cell derived membranous nanoparticles. EVs are important mediators of cell-cell communication via the transfer of bioactive content and as such they are being investigated for disease diagnostics as biomarkers and for potential therapeutic cargo delivery to recipient cells. However, existing methods for isolating EVs from biological samples suffer from challenges related to co-isolation of unwanted materials such as proteins, nucleic acids, and lipoproteins. In the pursuit of improved EV isolation techniques, we introduce multimodal flowthrough chromatography (MFC) as a scalable alternative to size exclusion chromatography (SEC). The use of MFC offers significant advantages for purifying EVs, resulting in enhanced yields and increased purity with respect to protein and nucleic acid co-isolates from conditioned 3D cell culture media. Compared to SEC, significantly higher EV yields with similar purity and preserved functionality were also obtained with MFC in 2D cell cultures. Additionally, MFC yielded EVs from serum with comparable purity to SEC and similar apolipoprotein B content. Overall, MFC presents an advancement in EV purification yielding EVs with high recovery, purity, and functionality, and offers an accessible improvement to researchers currently employing SEC., Competing Interests: PV serves on the scientific advisory board of Evox Therapeutics Ltd. SEB has equity and interests in Evox Therapeutics Ltd, and has held an employment contract with Evox Therapeutics Ltd., Oxford, UK, during the completion of this article. MJAW is a cofounder of Evox Therapeutics, Pepgen, Orfonyx and Isogenix. MJAW is a scientific advisory board member of Evox Therapeutics and Pepgen, and is a Director of Oxford University Innovation. The University of Oxford has patents issued and/or pending for oligonucleotide technologies and delivery technologies (on which MJAW is an inventor). Evox Therapeutics, Pepgen, Orfonyx and Isogenix had no influence on the conception or realisation of this article., (© 2024 The Authors. Journal of Extracellular Biology published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)

The loss-of-function of the proprotein convertase subtilisin-kexin type 9 (Pcsk9) gene has been associated with significant reductions in plasma serum low-density lipoprotein cholesterol (LDL-C) levels. Both CRISPR/Cas9 and CRISPR-based editor-mediated Pcsk9 inactivation have successfully lowered plasma LDL-C and PCSK9 levels in preclinical models. Despite the promising preclinical results, these studies did not report how vehicle-mediated CRISPR delivery inactivating Pcsk9 affected low-density lipoprotein receptor recycling in vitro or ex vivo. Extracellular vesicles (EVs) have shown promise as a biocompatible delivery vehicle, and CRISPR/Cas9 ribonucleoprotein (RNP) has been demonstrated to mediate safe genome editing. Therefore, we investigated EV-mediated RNP targeting of the Pcsk9 gene ex vivo in primary mouse hepatocytes. We engineered EVs with the rapamycin-interacting heterodimer FK506-binding protein (FKBP12) to contain its binding partner, the T82L mutant FKBP12-rapamycin binding (FRB) domain, fused to the Cas9 protein. By integrating the vesicular stomatitis virus glycoprotein on the EV membrane, the engineered Cas9 EVs were used for intracellular CRISPR/Cas9 RNP delivery, achieving genome editing with an efficacy of ±28.1% in Cas9 stoplight reporter cells. Administration of Cas9 EVs in mouse hepatocytes successfully inactivated the Pcsk9 gene, leading to a reduction in Pcsk9 mRNA and increased uptake of the low-density lipoprotein receptor and LDL-C. These readouts can be used in future experiments to assess the efficacy of vehicle-mediated delivery of genome editing technologies targeting Pcsk9. The ex vivo data could be a step towards reducing animal testing and serve as a precursor to future in vivo studies for EV-mediated CRISPR/Cas9 RNP delivery targeting Pcsk9., (© 2024 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)

Cardiac progenitor cell (CPC)-derived small extracellular vesicles (sEVs) exhibit great potential to stimulate cardiac repair. However, the multifaceted nature of sEV heterogeneity presents a challenge in understanding the distinct mechanisms underlying their regenerative abilities. Here, a dual-step multimodal flowthrough and size-exclusion chromatography method was applied to isolate and separate CPC-derived sEV subpopulations to study the functional differences related to cardiac repair responses. Three distinct sEV subpopulations were identified with unique protein profiles. Functional cell assays for cardiac repair-related processes demonstrated that the middle-sized and smallest-sized sEV subpopulations exhibited the highest pro-angiogenic and anti-fibrotic activities. Proteasome activity was uniquely seen in the smallest-sized subpopulation. The largest-sized subpopulation showed no effect in any of the functional assays. This research uncovers the existence of sEV subpopulations, each characterized by a distinct composition and biological function. Enhancing our understanding of sEV heterogeneity will provide valuable insights into sEV mechanisms of action, ultimately accelerating the translation of sEV therapeutics., (© 2024 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals LLC on behalf of International Society for Extracellular Vesicles.)