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This opinion article highlights the critical role of laboratory medicine and emerging technologies in cardiovascular risk reduction through exposome analysis. The exposome encompasses all external and internal exposures an individual faces throughout their life, influencing the onset and progression of cardiovascular diseases (CVD). Integrating exposome data with genetic information allows for a comprehensive understanding of the multifactorial causes of CVD, facilitating targeted preventive interventions. Laboratory medicine, enhanced by advanced technologies such as metabolomics and artificial intelligence (AI), plays a pivotal role in identifying and mitigating these exposures. Metabolomics provides detailed insights into metabolic changes triggered by environmental factors, while AI efficiently processes complex datasets to uncover patterns and associations. This integration fosters a proactive approach in public health and personalized medicine, enabling earlier detection and intervention. The article calls for global implementation of exposome technologies to improve population health, emphasizing the need for robust technological platforms and policy-driven initiatives to seamlessly integrate environmental data with clinical diagnostics. By harnessing these innovative technologies, laboratory medicine can significantly contribute to reducing the global burden of cardiovascular diseases through precise and personalized risk mitigation strategies., (© 2024 Walter de Gruyter GmbH, Berlin/Boston.)

CD8 T cells are the predominant effector cells of adaptive immunity in preventing cytomegalovirus (CMV) multiple-organ disease caused by cytopathogenic tissue infection. The mechanism by which CMV-specific, naïve CD8 T cells become primed and clonally expand is of fundamental importance for our understanding of CMV immune control. For CD8 T-cell priming, two pathways have been identified: direct antigen presentation by infected professional antigen-presenting cells (pAPCs) and antigen cross-presentation by uninfected pAPCs that take up antigenic material derived from infected tissue cells. Studies in mouse models using murine CMV (mCMV) and precluding either pathway genetically or experimentally have shown that, in principle, both pathways can congruently generate the mouse MHC/H-2 class-I-determined epitope-specificity repertoire of the CD8 T-cell response. Recent studies, however, have shown that direct antigen presentation is the canonical pathway when both are accessible. This raised the question of why antigen cross-presentation is ineffective even under conditions of high virus replication thought to provide high amounts of antigenic material for feeding cross-presenting pAPCs. As delivery of antigenic material for cross-presentation is associated with programmed cell death, and as CMVs encode inhibitors of different cell death pathways, we pursued the idea that these inhibitors restrict antigen delivery and thus CD8 T-cell priming by cross-presentation. To test this hypothesis, we compared the CD8 T-cell responses to recombinant mCMVs lacking expression of the apoptosis-inhibiting protein M36 or the necroptosis-inhibiting protein M45 with responses to wild-type mCMV and revertant viruses expressing the respective cell death inhibitors. The data reveal that increased programmed cell death improves CD8 T-cell priming in mice capable of antigen cross-presentation but not in a mutant mouse strain unable to cross-present. These findings strongly support the conclusion that CMV cell death inhibitors restrict the priming of CD8 T cells by antigen cross-presentation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ebert et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A fundamental task for the brain is to generate predictions of future sensory inputs, and signal errors in these predictions. Many neurons have been shown to signal omitted stimuli during periodic stimulation, even in the retina. However, the mechanisms of this error signaling are unclear. Here we show that depressing inhibitory synapses shape the timing of the response to an omitted stimulus in the retina. While ganglion cells, the retinal output, responded to an omitted flash with a constant latency over many frequencies of the flash sequence, we found that this was not the case once inhibition was blocked. We built a simple circuit model and showed that depressing inhibitory synapses were a necessary component to reproduce our experimental findings. A new prediction of our model is that the accuracy of the constant latency requires a sufficient amount of flashes in the stimulus, which we could confirm experimentally. Depressing inhibitory synapses could thus be a key component to generate the predictive responses observed in the retina, and potentially in many brain areas., (© 2024. The Author(s).)

Next to its classical role in MHC II-mediated antigen presentation, CD74 was identified as a high-affinity receptor for macrophage migration inhibitory factor (MIF), a pleiotropic cytokine and major determinant of various acute and chronic inflammatory conditions, cardiovascular diseases and cancer. Recent evidence suggests that CD74 is expressed in T cells, but the functional relevance of this observation is poorly understood. Here, we characterized the regulation of CD74 expression and that of the MIF chemokine receptors during activation of human CD4 + T cells and studied links to MIF-induced T-cell migration, function, and COVID-19 disease stage. MIF receptor profiling of resting primary human CD4 + T cells via flow cytometry revealed high surface expression of CXCR4, while CD74, CXCR2 and ACKR3/CXCR7 were not measurably expressed. However, CD4 + T cells constitutively expressed CD74 intracellularly, which upon T-cell activation was significantly upregulated, post-translationally modified by chondroitin sulfate and could be detected on the cell surface, as determined by flow cytometry, Western blot, immunohistochemistry, and re-analysis of available RNA-sequencing and proteomic data sets. Applying 3D-matrix-based live cell-imaging and receptor pathway-specific inhibitors, we determined a causal involvement of CD74 and CXCR4 in MIF-induced CD4 + T-cell migration. Mechanistically, proximity ligation assay visualized CD74/CXCR4 heterocomplexes on activated CD4 + T cells, which were significantly diminished after MIF treatment, pointing towards a MIF-mediated internalization process. Lastly, in a cohort of 30 COVID-19 patients, CD74 surface expression was found to be significantly upregulated on CD4 + and CD8 + T cells in patients with severe compared to patients with only mild disease course. Together, our study characterizes the MIF receptor network in the course of T-cell activation and reveals CD74 as a novel functional MIF receptor and MHC II-independent activation marker of primary human CD4 + T cells., (© 2024. The Author(s).)

N-glycosylation of the Fc part is a (critical) quality attribute of therapeutic antibodies and Fc-containing biotherapeutics, that impacts their stability, immunogenicity, pharmacokinetics, and effector functions. Current glycosylation analysis methods focus on the absolute amounts of glycans, neglecting the apparent glycan distribution over the entirety of proteins. The combination of the two Fc N-glycans, herein referred to as glyco-pair, therefore remains unknown, which is a major drawback for N-glycan impact assessment. This study presents a comprehensive workflow for the analysis and characterization of Fc N-glycan pairing in biotherapeutics, addressing the limitations of current glycosylation analysis methods. The applicability of the method across various biotherapeutic proteins including antibodies, bispecific antibody formats, and a Fc-Fusion protein is demonstrated, and the impact of method conditions on glycan pairing analysis is highlighted. Moreover, the influence of the molecular format, Fc backbone, production process, and cell line on glycan pairing pattern was investigated. The results underscore the significance of comprehensive glycan pairing analysis to accurately assess the impact of N-glycans on important product quality attributes of therapeutic antibodies and Fc-containing biotherapeutics., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Maximilian Meudt, Julia Baumeister, Michaela Blech and Fabian Higel are employees of Boehringer Ingelheim GmbH & Co. KG, which is developing, manufacturing, and marketing biopharmaceutical products., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: The Rural Surgical Obstetrical Networks (RSON) initiative in BC was developed to stabilize and grow low volume rural surgical and obstetrical services. One of the wrap-around supportive interventions was funding for Continuous Quality Improvement (CQI) initiatives, done through a local provider-driven lens. This paper reviews mixed-methods findings on providers' experiences with CQI and the implications for service stability., Background: Small, rural hospitals face barriers in implementing quality improvement initiatives due primarily to lack of resource capacity and the need to prioritize clinical care when allocating limited health human resources. Given this, funding and resources for CQI were key enablers of the RSON initiative and seen as an essential part of a response to assuaging concerns of specialists at higher volume sites regarding quality in lower volume settings., Methods: Data were derived from two datasets: in-depth, qualitative interviews with rural health care providers and administrators over the course of the RSON initiative and through a survey administered at RSON sites in 2023., Findings: Qualitative findings revealed participants' perceptions of the value of CQI (including developing expanded skillsets and improved team function and culture), enablers (the organizational infrastructure for CQI projects), challenges in implementation (complications in protecting/prioritizing CQI time and difficulty with staff engagement) and the importance of local leadership. Survey findings showed high ratings for elements of team function that relate directly to CQI (team process and relationships)., Conclusion: Attention to effective mechanisms of CQI through a rural lens is essential to ensure that initiatives meet the contextual realities of low-volume sites. Instituting pathways for locally-driven quality improvement initiatives enhances team function at rural hospitals through creating opportunities for trust building and goal setting, improving communication and increasing individual and team-wide motivation to improve patient care., Competing Interests: Sean Ebert received sessional funding from the Rural Coordination Centre of BC (RCCbc) in his role as medical lead for the Quality Improvement Pillar for the Rural Surgical and Obstetrical Networks (RSON) initiative. In his capacity as medical lead he received funding to travel to the rural communities where the participants we report on in this manuscript work and provide care. Tom Skinner is employed by the Rural Coordination Centre of BC (RCCbc) as the Project Manager of the RSON Initiative. The findings reported in the survey are part of the evaluation of RSON. As an employee, his travel to the rural communities were covered because he was supporting hospital teams in implementing the PROES survey and interpreting the result. Kim Williams received salary funding from the Rural Coordination Centre of BC (RCCbc) in her capacity as administrative co-lead for the Rural Surgical and Obstetrical Networks (RSON) initiative. She received funding to travel to the rural communities where the participants whom we report on in this manuscript work and provide care. Nancy Humber received funding from the Rural Coordination Centre of BC (RCCbc) in her role as a clinical lead for the Rural Surgical Obstetrical Network. In her capacity as a clinical lead, she received funding for travel to the rural communities where the participants whom we report on in this manuscript work and provide care. All other authors are part of the RSON evaluation team at the University of British Columbia, Canada and have no conflicts of interest to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Kornelsen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: The number of hospitalized patients with severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) does not differentiate between patients admitted due to coronavirus disease 2019 (COVID-19) (ie, primary cases) and incidental SARS-CoV-2 infection (ie, incidental cases). We developed an adaptable method to distinguish primary cases from incidental cases upon hospital admission., Design: Retrospective cohort study., Setting: Data were obtained from 3 German tertiary-care hospitals., Patients: The study included patients of all ages who tested positive for SARS-CoV-2 by a standard quantitative reverse-transcription polymerase chain reaction (RT-PCR) assay upon admission between January and June 2022., Methods: We present 2 distinct models: (1) a point-of-care model that can be used shortly after admission based on a limited range of parameters and (2) a more extended point-of-care model based on parameters that are available within the first 24-48 hours after admission. We used regression and tree-based classification models with internal and external validation., Results: In total, 1,150 patients were included (mean age, 49.5±28.5 years; 46% female; 40% primary cases). Both point-of-care models showed good discrimination with area under the curve (AUC) values of 0.80 and 0.87, respectively. As main predictors, we used admission diagnosis codes (ICD-10-GM), ward of admission, and for the extended model, we included viral load, need for oxygen, leucocyte count, and C-reactive protein., Conclusions: We propose 2 predictive algorithms based on routine clinical data that differentiate primary COVID-19 from incidental SARS-CoV-2 infection. These algorithms can provide a precise surveillance tool that can contribute to pandemic preparedness. They can easily be modified to be used in future pandemic, epidemic, and endemic situations all over the world.

The self-prioritization effect suggests that self-relevant information has a processing advantage over information that is not directly associated with the self. In consequence, reaction times are faster and accuracy rates higher when reacting to self-associated stimuli rather than to other-related stimuli (Sui et al., Journal of Experimental Psychology: Human Perception and Performance 38:1105-1117, 2012). This spurs the assumption that self-associated action-effects should also be perceived earlier than other-related outcomes. One way to measure this is temporal binding. Previous research indeed showed that the perceived temporal interval between actions and self-associated outcomes was reduced compared to friend- and other-associated outcomes. However, the employed method (interval estimations) and several experimental design choices make it impossible to discern whether the perceived shortening of the interval between a keypress and a self-relevant outcome is due to a perceptual shift of the action or of the action-effect or both. Thus, we conducted four experiments to assess whether temporal binding can indeed be modulated by self-relevance and if so where this perceptual bias is located. The results did not support stronger temporal binding for self- vs other-related action-effects. We discuss these results against the backdrop of the attentional basis of self-prioritization and propose directions for future research., (© 2024. The Author(s).)

Introduction: Rural patients face barriers to accessing surgical care and often need to travel long distance for pre- or post-surgical consultations. Although adaptation to the COVID-19 pandemic has demonstrated the efficacy of virtual care, there is minimal data available to evaluate patient satisfaction with this modality and consequent health service utilization if virtual services are not available., Methods: An online survey was conducted with participants living in rural British Columbia, Canada who had undergone surgery within 12 months of data collection and had either virtual or face-to-face pre- or post-surgical consultations. It was supplemented by an in-person survey administered in two rural sites to all patients who had a virtual visit prior to undergoing procedural care. A ten-point scale was used to assess satisfaction. Quantitative and qualitative data were collected and analyzed., Results: Findings from the province-wide survey (n = 163) revealed no significant differences in average satisfaction ratings between people with in-person and virtual surgical consultations (8.03 versus 8.38, p  = 0.26). However, most participants indicated that virtual appointments saved them time traveling, energy, and money and made them less dependent on others, accruing significant social benefit.In the community-focused sample (n = 71), 38% said they would not have had the procedure without a virtual visit option and 21% said that they would have delayed the procedure. Virtual consultations saved patients an average of 9 h (range 1-90). Participants traveled an average of 427 kilometers round trip to have the procedures., Conclusion: Findings reveal costs and time saved in accessing care due to the introduction of pre- and post-operative virtual care visits, and further investments in virtual care are warranted. This will contribute to promoting equitable access to healthcare for rural residents., Competing Interests: –Dr Sean Ebert received sessional funding from the Rural Coordination Centre of BC (RCCbc) in his role as medical lead for the Quality Improvement Pillar for the RSON initiative. In his capacity as medical lead he received funding to travel to the rural communities where the patients whom we report on in this manuscript had surgical care.–Tom Skinner is employed by the Rural Coordination Centre of BC (RCCbc) as the Project Manager of the RSON Initiative. The findings reported in the community survey are part of the evaluation of RSON. As an employee, his travel to the rural communities was covered because he was supporting hospital teams in implementing the PROES survey and interpreting the result.–All other authors are part of the RSON evaluation team at the University of British Columbia, Canada and have no conflicts of interest to declare., (© The Author(s) 2024.)

Introduction: Outreach care has long been used in Canada to address the lack of access to specialist care in rural settings, but research on the experiences of specialists providing these services is lacking. This descriptive survey study aimed to understand 1) specialists' motivation for engaging in outreach work, (2) their perceptions of the quality of care at their rural outreach hospital, and (3) the supports they receive for their outreach work, in order to create a supportive framework to encourage specialist outreach contributions., Methods: In July 2022, specialist physicians who provide outreach operating room services at rural hospitals participating in the Rural Surgical and Obstetrical Networks initiative in the province of British Columbia were invited to complete an anonymous survey., Results: 21 of 45 invited outreach specialists completed the survey (47% response rate). Three-quarters of respondents had a surgical specialty. The opportunity to deliver care to underserved patients was the most common motivator for outreach work. Rural hospitals received high ratings from respondents on overall safety and various aspects of communication and teamwork. Postoperative care was a concern for a minority (one-fifth) of respondents, and about half had experienced unnecessary delays between procedures some or most of the time. Generally, respondents felt integrated into rural teams and reported receiving adequate nursing and anesthetic support. The two most common desired additional supports were better/more equipment and space and additional staffing. All 19 respondents not planning to retire soon intended to provide outreach services for at least three more years., Conclusion: Specialists providing outreach OR services in small volume rural hospitals in BC usually have altruistic motives for outreach work. For the most part, these specialists have positive experiences in rural hospitals, but they can be better supported through investment in infrastructure and health human resources. Specialists intend to provide outreach services long-term, indicating a stable outreach workforce. More research on the facilitators and barriers of specialist outreach work is needed., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Parajulee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The prognostic impact of PICALM::MLLT10 status in childhood leukaemia is not well described. Ten International Berlin Frankfurt Münster-affiliated study groups and the Children's Oncology Group collaborated in this multicentre retrospective study. The presence of the PICALM::MLLT10 fusion gene was confirmed by fluorescence in situ hybridization and/or RNA sequencing at participating sites. Ninety-eight children met the study criteria. T-cell acute lymphoblastic leukaemia (T-ALL) and acute myeloid leukaemia (AML) predominated 55 (56%) and 39 (40%) patients, respectively. Most patients received a chemotherapy regimen per their disease phenotype: 58% received an ALL regimen, 40% an AML regimen and 1% a hybrid regimen. Outcomes for children with PICALM::MLLT10 ALL were reasonable: 5-year event-free survival (EFS) 67% and 5-year overall survival (OS) 76%, but children with PICALM::MLLT10 AML had poor outcomes: 5-year EFS 22% and 5-year OS 26%. Haematopoietic stem cell transplant (HSCT) did not result in a significant improvement in outcomes for PICALM::MLLT10 AML: 5-year EFS 20% for those who received HSCT versus 23% for those who did not (p = 0.6) and 5-year OS 37% versus 36% (p = 0.7). In summary, this study confirms that PICALM::MLLT10 AML is associated with a dismal prognosis and patients cannot be salvaged with HSCT; exploration of novel therapeutic options is warranted., (© 2023 British Society for Haematology and John Wiley & Sons Ltd.)

The current trend in biopharmaceutical drug manufacturing is towards increasing potency and complexity of products such as peptide scaffolds, oligonucleotides and many more. Therefore, a universal affinity purification step is important in order to meet the requirements for cost and time efficient drug production. By using a self-splicing intein affinity tag, a purification template is generated that allows for a universal chromatographic affinity capture step to generate a tagless target protein without the use of proteases for further tag removal. This study describes the successful implementation of gp41-1-based split inteins in a chromatographic purification process for, e.g., E. coli -derived targets. The tagless target is generated in a single-step purification run. The on-column cleavage is induced by triggering a simple pH change in the buffer conditions without the need for additives such as Zn 2+ or thiols. This system has proven to be reusable for at least ten purification cycles that use 150 mM H 3 PO 4 as the cleaning agent., Competing Interests: The authors declare that this study received funding from Merck – Life Science KGaA, Darmstadt, Germany. The funder had the following involvement in the study: financial support for the research, authorship, and/or publication of this article. Authors MK, VK, TB, and OR were employed by Merck—Life Science KGaA. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Knapp, Kohl, Best, Rammo and Ebert.)

During the development process of therapeutic monoclonal antibodies (mAbs), it is crucial to control (critical) quality attributes such as N-glycosylation influencing pharmacokinetics (PK) and Fc effector functions. Previous reports have shown that mAbs containing high-mannose N-glycans are cleared faster from blood circulation, leading to reduced half-lives. The high-mannose N-glycan content of mAbs can be influenced during the cell culture process by factors such as cell lines, process conditions, and media. Furthermore, mAbs have either one high mannose N-glycan (asymmetrical high-mannose glyco-pair) or two high mannose N-glycans (symmetrical high-mannose glyco-pair). The hypothesis that the mannose receptor (MR, CD206) accelerates clearance by facilitating their internalization and subsequent lysosomal degradation is widespread. However, the interaction between MR and mAbs has not been explicitly demonstrated. This study aimed to investigate this interaction, providing the first systematic demonstration of MR binding to the Fc region of mAbs with high-mannose N-glycans. Two novel analytical methods, MR surface plasmon resonance and MR affinity chromatography, were developed and applied to investigate the MR-mAb interaction. The interaction is found to be dependent on high-mannose content, but is independent of the mAb format or sequence. However, different glyco-pairs exhibited varying binding affinities to the MR, with the symmetrical high-mannose glyco-pair showing the strongest binding properties. These findings strengthen the hypothesis for the MR-mediated mAb interaction and contribute to a deeper understanding of the MR-mAb interaction, which could affect the criticality of high-mannose containing mAbs development strategies of IgG-based molecules and improve their PK profiles.

The increasing complexity and volume of mass spectrometry (MS) data have presented new challenges and opportunities for proteomics data analysis and interpretation. In this chapter, we provide a comprehensive guide to transforming MS data for machine learning (ML) training, inference, and applications. The chapter is organized into three parts. The first part describes the data analysis needed for MS-based experiments and a general introduction to our deep learning model SpeCollate-which we will use throughout the chapter for illustration. The second part of the chapter explores the transformation of MS data for inference, providing a step-by-step guide for users to deduce peptides from their MS data. This section aims to bridge the gap between data acquisition and practical applications by detailing the necessary steps for data preparation and interpretation. In the final part, we present a demonstrative example of SpeCollate, a deep learning-based peptide database search engine that overcomes the problems of simplistic simulation of theoretical spectra and heuristic scoring functions for peptide-spectrum matches by generating joint embeddings for spectra and peptides. SpeCollate is a user-friendly tool with an intuitive command-line interface to perform the search, showcasing the effectiveness of the techniques and methodologies discussed in the earlier sections and highlighting the potential of machine learning in the context of mass spectrometry data analysis. By offering a comprehensive overview of data transformation, inference, and ML model applications for mass spectrometry, this chapter aims to empower researchers and practitioners in leveraging the power of machine learning to unlock novel insights and drive innovation in the field of mass spectrometry-based omics., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: The Rural Surgical Obstetrical Networks (RSON) project was developed in response to the persistent attrition of rural maternity services across Canada over the past two decades. While other research has demonstrated the adverse health and psychosocial consequences of losing local maternity services, this paper explores the impact of a program designed to increase the sustainability of rural services themselves, through the funding of four "pillars": increased scope and volume, clinical coaching, continuous quality improvement (CQI) and remote presence technology., Methods: We conducted in-depth, qualitative research interviews with rural health care providers and administrators in eight rural communities across British Columbia to understand the impact of the RSON program on maternity services. Researchers used thematic analysis to generate common themes across the dataset and interpret findings., Findings: Participants articulated six themes regarding the sustainability of maternity care as actualized through the RSON project: safety and quality through quality improvement opportunities, improved access to care through increased surgical volume and OR backup, optimized team function through innovative models of care, improved infrastructure, local innovation surrounding workforce shortages, and locally tailored funding models., Conclusion: Rural maternity sites benefited from the funding offered through the RSON pillars, as demonstrated by larger volumes of local deliveries, nearly unanimous positive accounts of the interventions by health care providers, and evidence of staffing stability during the study time frame. As such, the interventions provided through the Rural Surgical Obstetrical Networks project as well as study findings on the common themes of sustainable maternity care should be considered when planning core rural health services funding schemes., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Tissue inhibitor of metalloproteinases-1 (TIMP-1), an important regulator of matrix metalloproteinases (MMPs), has recently been shown to interact with CD74, a receptor for macrophage migration inhibitory factor (MIF). However, the biological effects mediated by TIMP-1 through CD74 remain largely unexplored. Using sequence alignment and in silico protein-protein docking analysis, we demonstrated that TIMP-1 shares residues with both MIF and MIF-2, crucial for CD74 binding, but not for CXCR4. Subcellular colocalization, immunoprecipitation, and internalization experiments supported these findings, demonstrating that TIMP-1 interacts with surface-expressed CD74, resulting in its internalization in a dose-dependent manner, as well as with a soluble CD74 ectodomain fragment (sCD74). This prompted us to study the effects of the TIMP-1-CD74 axis on monocytes and vascular smooth muscle cells (VSCMs) to assess its impact on vascular inflammation. A phospho-kinase array revealed the activation of serine/threonine kinases by TIMP-1 in THP-1 pre-monocytes, in particular AKT. Similarly, TIMP-1 dose-dependently triggered the phosphorylation of AKT and ERK1/2 in primary human monocytes. Importantly, Transwell migration, 3D-based Chemotaxis, and flow adhesion assays demonstrated that TIMP-1 engagement of CD74 strongly promotes the recruitment response of primary human monocytes, while live cell imaging studies revealed a profound activating effect on VSMC proliferation. Finally, re-analysis of scRNA-seq data highlighted the expression patterns of TIMP-1 and CD74 in human atherosclerotic lesions, thus, together with our experimental data, indicating a role for the TIMP-1-CD74 axis in vascular inflammation and atherosclerosis.

Background: Melanoma is a lethal skin cancer, and the risk of developing it is increased by exposure to ultraviolet (UV) radiation. The production of cytokines such as interleukin-15 (IL-15), induced by the exposure of skin cells to UV rays, could also promote melanoma development. The aim of this study is to investigate the possible role of Interleukin-15/Interleukin-15 Receptor α (IL-15/IL-15Rα) complexes in melanoma development., Methods: The expression of IL-15/IL-15Rα complexes by melanoma cells was evaluated both ex vivo and in vitro by tissue microarray, PCR, and flow cytometry. The presence of the soluble complex (sIL-15/IL-15Rα) in the plasma of metastatic melanoma patients was detected using an ELISA assay. Subsequently, we investigated the impact of natural killer (NK) cell activation after rIL-2 starvation followed by exposure to the sIL-15/IL-15Rα complex. Finally, by analyzing public datasets, we studied the correlation between IL-15 and IL-15Rα expressions and melanoma stage, NK and T-cell markers, and overall survival (OS)., Results: Analysis of a melanoma tissue microarray shows a significant increase in the number of IL-15 + tumor cells from the benign nevi to metastatic melanoma stages. Metastatic melanoma cell lines express a phorbol-12-myristate-13-acetate (PMA)-cleavable membrane-bound IL-15 (mbIL-15), whereas cultures from primary melanomas express a PMA-resistant isoform. Further analysis revealed that 26% of metastatic patients present with consistently high plasmatic levels of sIL-15/IL-15Rα. When the recombinant soluble human IL-15/IL-15Rα complex is added to briefly starved rIL-2-expanded NK cells, these cells exhibit strongly reduced proliferation and levels of cytotoxic activity against K-562 and NALM-18 target cells. The analysis of public gene expression datasets revealed that high IL-15 and IL-15Rα intra-tumoral production correlates with the high levels of expression of CD5 + and NKp46 + (T and NK markers) and significantly correlates with a better OS in stages II and III, but not in stage IV., Conclusions: Membrane-bound and secreted IL-15/IL-15Rα complexes are continuously present during progression in melanoma. It is notable that, although IL-15/IL-15Rα initially promoted the production of cytotoxic T and NK cells, at stage IV promotion of the development of anergic and dysfunctional cytotoxic NK cells was observed. In a subgroup of melanoma metastatic patients, the continuous secretion of high amounts of the soluble complex could represent a novel NK cell immune escape mechanism., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Di Matteo, Munari, Fiore, Santopolo, Sampaoli, Pelosi, Chouaib, Tumino, Vacca, Mariotti, Ebert, Machwirth, Haas, Pezzullo, Pietra, Grottoli, Buart, Mortier, Maggi, Moretta, Caruana and Azzarone.)

The metabolic state represents a major hurdle for an effective adoptive T cell therapy (ACT). Indeed, specific lipids can harm CD8 + T cell (CTL) mitochondrial integrity, leading to defective antitumor responses. However, the extent to which lipids can affect the CTL functions and fate remains unexplored. Here, we show that linoleic acid (LA) is a major positive regulator of CTL activity by improving metabolic fitness, preventing exhaustion, and stimulating a memory-like phenotype with superior effector functions. We report that LA treatment enhances the formation of ER-mitochondria contacts (MERC), which in turn promotes calcium (Ca 2+ ) signaling, mitochondrial energetics, and CTL effector functions. As a direct consequence, the antitumor potency of LA-instructed CD8 T cells is superior in vitro and in vivo. We thus propose LA treatment as an ACT potentiator in tumor therapy., Competing Interests: Declaration of interests C.B.N.L., L.N., and T.M. reported a patent application related to this study. P.D.G. was a scientific consultant for Juno Therapeutics., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

So-called meat hybrids are a new class of products where a fraction of the meat product (e.g., 20%) is replaced with alternative protein sources, such as plant-based ones. Research suggests that these products could serve as a low-threshold offer for a specific target group that wants to cut down on meat, thereby facilitating the transition toward a more healthy and sustainable diet. Nonetheless, data demonstrate that meat hybrids with a high substantial meat substitution level often fail in the market. This study summarises findings on the physicochemical properties, sensory, and acceptance of six different meat hybrids (70% meat and 30% plant proteins) that were collected in the framework of a case study in the project AiF 196 EN. For this purpose, sensory characteristics were collected via two QDA sessions and a hedonic consumer test. Furthermore, the hybrid recipes were analysed in their proximate composition. The respective recipes varied in protein source (soybean, pumpkin, and pea) and mode of incorporation [textured vegetable protein (TVP), high moisture extrudate (HME)]. It was shown that a meat hybrid with a relatively high share of 30% plant-based proteins with peas as a protein source and TVP as a processing method can still attract consumers., Competing Interests: AP was employed by Prokribus GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Baune, Broucke, Ebert, Gibis, Weiss, Enneking, Profeta, Terjung and Heinz.)

In spite of its central role in biology and disease, protein turnover is a largely understudied aspect of most proteomic studies due to the complexity of computational workflows that analyze in vivo turnover rates. To address this need, we developed a new computational tool, TurnoveR, to accurately calculate protein turnover rates from mass spectrometric analysis of metabolic labeling experiments in Skyline, a free and open-source proteomics software platform. TurnoveR is a straightforward graphical interface that enables seamless integration of protein turnover analysis into a traditional proteomics workflow in Skyline, allowing users to take advantage of the advanced and flexible data visualization and curation features built into the software. The computational pipeline of TurnoveR performs critical steps to determine protein turnover rates, including isotopologue demultiplexing, precursor-pool correction, statistical analysis, and generation of data reports and visualizations. This workflow is compatible with many mass spectrometric platforms and recapitulates turnover rates and differential changes in turnover rates between treatment groups calculated in previous studies. We expect that the addition of TurnoveR to the widely used Skyline proteomics software will facilitate wider utilization of protein turnover analysis in highly relevant biological models, including aging, neurodegeneration, and skeletal muscle atrophy.

Psychology has primarily studied hope as a value-neutral trait even though it has a history of being counted among the virtues. The current study seeks to conceptualize hope as a virtue while building on the dominant empirical paradigm (Hope Theory; Snyder, 2002). Ithemba is the isiZulu word for hope, and this qualitative study investigated the lived experiences and meaning construction of ithemba/hope among 13 nominated moral exemplars in a South African township. Participants ranged from 20 to 75+, including farmers, educators, caregivers, entrepreneurs, and construction workers. Independent coders conducted thematic analysis from a theoretical top-down process (within Hope Theory and virtue science frameworks) and an inductive bottom-up approach (open coding). Data reflected much of the existing Hope Theory model; however, many pathways were relational and spiritual, and goals were inherently beneficial to others. A virtue science framework was used to construct the cognitive, motivational, and behavioral dimensions of virtuous hope. This study constructed the operational definition of virtuous hope as the ardent pursuit of realizing a particular vision of the common good with intention and action, often growing out of adversity and shaped in relation to other people and the transcendent., Supplementary Information: The online version contains supplementary material available at 10.1007/s41042-022-00083-1., Competing Interests: Conflict of interestThe authors report there are no competing interests to declare and no financial information to report. Partial funding for interview costs (transportation, transcription, translation) was provided from an internal research grant from the University of Indianapolis InQuery Grant., (© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

The visible non-verbal behavior of volleyball players after a point might indicate their affective state. The purpose of the present study was to investigate whether the point outcome (i.e., win vs. loss) and the importance of the situation (i.e., first vs. fifth set) influence how well the affective states (indexed through non-verbal behavior) can be recognized. For this purpose, 111 participants were presented 60 video excerpts showing the non-verbal behavior of volleyball teams immediately after a rally and asked whether the shown team had just won or lost a point. The results indicated that the recognition rate was significantly higher for won (80.09%) than for lost points (65.56%) as well as for points of the fifth set (77.84%) than for points of the first set (68.51%). Furthermore, an explorative analysis showed that the recognition rate was significantly higher for won points of the fifth set (89.19%) than for those of the first set (72.13%)., Competing Interests: Declaration of competing interest The authors declare that there are no competing financial interest or personal relationships that influenced the work reported in the paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: The prognosis of children with acute lymphoblastic leukemia (ALL) has improved considerably over the past five decades. However, to achieve cure in patients with refractory or relapsed disease, novel treatment options are necessary., Methods: In the multicenter trial Cooperative Study Group for Childhood Acute Lymphoblastic Leukemia (CoALL)08-09, one additional treatment element consisting of the rarely used chemotherapeutic agent amsacrine combined with etoposide and methylprednisolone (AEP) (amsacrine 2 × 100 mg/m 2 , etoposide 2 × 500 mg/m 2 , and methylprednisolone 4 × 1000 mg/m 2 ) was incorporated into the first-line treatment of pediatric patients with poor treatment responses at the end of induction (EOI), measured by minimal residual disease (MRD). These patients were stratified into a high-risk intensified arm (HR-I), including an AEP element at the end of consolidation. Patients with induction failure (IF), that is, with lack of cytomorphological remission EOI, were eligible for hematopoietic stem cell transplantation (HSCT) after remission had been reached. These patients received AEP as a part of their MRD-guided bridging-to-transplant treatments., Results: A significant improvement in probability of overall survival (pOS) was noted for the CoALL08-09 HR-I patients compared to MRD-matched patients from the preceding CoALL07-03 trial in the absence of severe or persistent treatment-related toxicities. Relapse rate and probability of event-free survival (pEFS) did not differ significantly between trials. In patients with IF, stable or improved MRD responses after AEP were observed without severe or persistent treatment-related toxicities., Conclusion: In conclusion, AEP is well tolerated as a component of the HR treatment and is useful in bridging-to-transplant settings., (© 2022 The Authors. Pediatric Blood & Cancer published by Wiley Periodicals LLC.)

Purpose: This retrospective cross-sectional cohort study investigated the influence of posture on lordosis (LL), length of the spinal canal (LSC), anteroposterior diameter (APD L1-L5), dural cross-sectional area (DCSA) of the lumbar spinal canal, and the prevalence of redundant nerve roots (RNR) using positional magnetic resonance imaging (MRI) (0.6 T)., Methods: Sixty-eight patients with single-level degenerative central lumbar spinal stenosis (cLSS) presenting with RNR in the standing position (STA) were also investigated in supine (SUP) or neutral seated (SIT) and flexed seated (FLEX) positions. Additionally, 45 patients complaining of back pain and without MRI evidence of LSS were evaluated. Statistical significance was set at p < 0.05., Results: Controls (A) and patients with cLSS (B) were comparable in terms of mean age (p = 0.88) and sex (p = 0.22). The progressive transition from STA to FLEX led to a comparable decrease in LL (p = 0.97), an increase in LSC (p = 0.80), and an increase in APD L1-L5 (p = 0.78). The APD of the stenotic level increased disproportionally between the different postures, up to 67% in FLEX compared to 29% in adjacent non-stenotic levels (p < 0.001). Therefore, the prevalence of RNR decreased to 49, 26, and 4% in SUP, SIT, and FLEX, respectively., Conclusion: The prevalence of RNR in standing position was underestimated by half in supine position. Body postures modified LL, LSC, and APD similarly in patients and controls. Stenotic levels compensated for insufficient intraspinal volume with a disproportionate enlargement when switching from the STA to FLEX., (© 2022. The Author(s).)

Refugees constitute a vulnerable group with an increased risk of developing trauma-related disorders. From a clinician's integrative perspective, navigating the detrimental impact of the social, economic, structural, and political factors on the mental health of refugees is a daily experience. Therefore, a collective effort must be made to reduce health inequities. The authors developed a treatment concept which provides broader care structures within a scientific practitioner's approach. The resulting "Trauma Network" addresses the structural challenges for refugees in Middle Hesse. Accompanying research provided a sound basis for further discussions with policy-makers to improve the situation for refugees in the short- and long-term.

Statistical body shape models (SBSM) provide compact, flexible representations of body shape that can be implemented in design software. However, few SBSMs have been created to represent adults in supported seated postures that are relevant for the design of seated environments, and none has incorporated the effects of age. This paper presents an SBSM based on surface laser-scan data from 155 U.S. adults. The data were processed to obtain homologous mesh structure and symmetric geometry, and the processed data were statistically analysed using principal component analysis to obtain a compact representation of the data variance. Regression analysis was conducted to predict body size and shape from stature, body mass index, ratio of sitting height to stature, sex, and age. The resulting model allows rapid generation of realistic body models for applications, including product design, accommodation assessment, and safety system optimisation. The model is publicly accessible at HumanShape.org. Practitioner summary: This paper presents a statistical model that represents adult body shapes in a supported seated posture based on 3 D anthropometric measurements. This model is the first whole-body parametric model known to incorporate age effects based on data extending beyond 65 years of age.

The increasing use of wet texturized plant proteins as meat substitutes requires a characterization of their functional properties, especially in terms of pH-behavior when being mixed with meat proteins to create so-called hybrid products. In this study, a minced model system containing pork meat, curing salt, and various amounts (0-100 wt%) of wet extruded proteins from pea (Pea I, II), pumpkin (Pumpkin I, II, III), and sunflower was used to evaluate the effect of mixing on pH and time-dependent pH-changes upon the addition of glucono-delta-lactone (GDL). Increasing concentrations of plant extrudates resulted in a linear increase of the initial (pH 0h ), intermediate (pH 6h ), and final pH 48h for all samples and higher slopes at higher native pH of extrudates were found. Acidification kinetics of all samples were similar with a distinct pH-drop by 0.3 to 0.8 pH-units per wt% GDL in the first 6 h, followed by a plateau where pH remained constant. At extrudate concentrations of 5 wt% (Pea I, II, Pumpkin I, II) or 15 wt% (Pumpkin III, Sunflower), a sufficient acidification with typically used GDL-amounts ( = 1 wt%) could be achieved, while higher plant protein contents required higher GDL-concentrations in order to reach a pH value of 5.0; a common target value in dry-cured sausages. A mathematical model was proposed to correlate pH, time, acidifier, extrudate concentration, and plant protein origin, to aid in the adjustment of dry-cured hybrid meat formulations, and to describe thresholds of the feasible extrudate and acidifier concentrations. PRACTICAL APPLICATION: Despite the increasing relevance of texturized plant proteins as meat mimetics, little is known about their functional and process-related properties. This study shows that plant protein origin, the level of meat replacement, and the amount of acidifier are linked to the time-dependent pH-value on the basis of a mathematical model. This brings food developers one step closer in creating tailored formulations and estimating the effects of these novel ingredients in the final product characteristics of hybrid meats and analogues., (© 2022 The Authors. Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Nanomaterials have emerged as an invaluable tool for the delivery of biomolecules such as DNA and RNA, with various applications in genetic engineering and post-transcriptional genetic manipulation. Alongside this development, there has been an increasing use of polymer-based techniques, such as polyethylenimine (PEI), to electrostatically load polynucleotide cargoes onto nanomaterial carriers. However, there remains a need to assess nanomaterial properties, conjugation conditions, and biocompatibility of these nanomaterial-polymer constructs, particularly for use in plant systems. In this work, we develop mechanisms to optimize DNA loading on single-walled carbon nanotubes (SWNTs) with a library of polymer-SWNT constructs and assess DNA loading ability, polydispersity, and both chemical and colloidal stability. Counterintuitively, we demonstrate that polymer hydrolysis from nanomaterial surfaces can occur depending on polymer properties and attachment chemistries, and we describe mitigation strategies against construct degradation. Given the growing interest in delivery applications in plant systems, we also assess the stress response of plants to polymer-based nanomaterials and provide recommendations for future design of nanomaterial-based polynucleotide delivery strategies.

Background: The use of plant proteins as food ingredients might be limited due to the presence of foreign or 'off' flavors, which may evolve during extraction and subsequent processing. In this study, the influence of dry (TVP) and wet (WTP) texturization on characteristic volatile compounds of two different pea protein isolates was assessed using gas chromatography-mass spectrometry-olfactometry (GC-MS-O) after direct immersion stir bar sorptive extraction (DI-SBSE)., Results: Twenty-four odor-active compounds were found, with a prevalence of carbonyls from fat oxidation. Nine of these compounds which are also known as major (off-) flavor contributors in peas were distinctively impacted in all texturates: hexanal, nonanal, 2-undecanone, (E)-2-octenal, (E, Z)-3,5-octadiene-2-one, (E, E)-2,4-decadienal, 2-pentyl-furan, 2-pentyl-pyridine, and γ-nonalactone. For example, hexanal, a characteristic green odorant, was reduced by up to sixfold by wet texturization, from 3.29 ± 1.05% (Pea Protein I) to 0.52 ± 0.02% (Pea WTP I). Furthermore, (E,Z)-3,5-Octadiene-2-one and (E,E)-2,4-decadienal were decreased by 1.5- and 1.8-fold when Pea Protein I and Pea TVP I were compared., Conclusion: An overall reduction in fat oxidation products and of green and fatty odor-active compounds was observed. The results represent a first insight into the process-related modulation of pea protein (off-) flavors to broaden the applicability of pea proteins as food ingredients., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

The exchange of animal-based for plant-based proteins is becoming more and more popular due to an increasing demand for alternative and more sustainable protein sources. In this study, solubilized water- (ws) or salt-and-water (sws) meat proteins were evaluated in their pH-dependent interactions with soluble protein fractions from wheat, pumpkin, sunflower, rapeseed, or potato proteins. For this purpose, 1 : 1 (v/v) mixtures of 1.0 wt% meat (ws or sws) and plant proteins were prepared at a sodium chloride concentration of 1.8 wt% (ionic strength: 0.31 mol L -1 ) and adjusted to different pH-values in between 4.5-7.0. While only slight differences were found upon comparison of interactions of ws and sws batches ( p > 0.05), interactions among these animal-based and soluble plant proteins took place. First, optical observations, light microscopy, and SDS-PAGE revealed increasing protein solubility with increasing pH. Second, particle size distributions (PSDs) revealed a shift towards slightly larger particle sizes e.g. at pH 5.3 and 7.0 with d 4,3 of 43.2 and 21.3 μm (sws) to 45.4 and 23.9 μm (sws + potato), respectively. Furthermore, heat-induced gel formation was improved at pH > 6.0, in particular in mixtures of meat and wheat or rapeseed proteins that formed a homogenous gel structure. Based on the obtained results, protein-protein complexations mainly by electrostatic forces are suggested which occur due to various pI of meat and plant proteins e.g. pH 7.5 (wheat), 7.2 (potato), and 6.6 (rapeseed) in comparison to 5.1 (ws) and 5.6 (sws). The filamentous microstructure of some gels (soluble fraction of rapeseed, potato and wheat proteins) led to the assumption that meat proteins, mainly at pH values greater than 5.8 (optimally ≥6.5), had a structuring effect on plant proteins.

Background: Agriculture faces significant global challenges including climate change and an increasing food demand due to a growing population. Addressing these challenges will require the adoption of transformative innovations into biotechnology practice, such as nanotechnology. Recently, nanomaterials have emerged as unmatched tools for their use as biosensors, or as biomolecule delivery vehicles. Despite their increasingly prolific use, plant-nanomaterial interactions remain poorly characterized, drawing into question the breadth of their utility and their broader environmental compatibility., Results: Herein, we characterize the response of Arabidopsis thaliana to single walled carbon nanotube (SWNT) exposure with two different surface chemistries commonly used for biosensing and nucleic acid delivery: oligonucleotide adsorbed-pristine SWNTs, and polyethyleneimine-SWNTs loaded with plasmid DNA (PEI-SWNTs), both introduced by leaf infiltration. We observed that pristine SWNTs elicit a mild stress response almost undistinguishable from the infiltration process, indicating that these nanomaterials are well-tolerated by the plant. However, PEI-SWNTs induce a much larger transcriptional reprogramming that involves stress, immunity, and senescence responses. PEI-SWNT-induced transcriptional profile is very similar to that of mutant plants displaying a constitutive immune response or treated with stress-priming agrochemicals. We selected molecular markers from our transcriptomic analysis and identified PEI as the main cause of this adverse reaction. We show that PEI-SWNT response is concentration-dependent and, when persistent over time, leads to cell death. We probed a panel of PEI variant-functionalized SWNTs across two plant species and identified biocompatible SWNT surface functionalizations., Conclusions: While SWNTs themselves are well tolerated by plants, SWNTs surface-functionalized with positively charged polymers become toxic and produce cell death. We use molecular markers to identify more biocompatible SWNT formulations. Our results highlight the importance of nanoparticle surface chemistry on their biocompatibility and will facilitate the use of functionalized nanomaterials for agricultural improvement., (© 2021. The Author(s).)

Background: Inappropriate prescribing of antibiotics has been identified as the most important modifiable risk factor for antimicrobial resistance., Objective: The purpose of this project was to improve guideline adherence and promote optimal use of outpatient antibiotics in the emergency department (ED)., Methods: Prescribing algorithms for community-acquired pneumonia (CAP), skin and soft tissue infections (SSTI), and urinary tract infections (UTI) were developed to integrate clinical practice guideline recommendations with local ED antibiogram data. Outcomes were evaluated through chart review of patients prescribed outpatient antibiotics by ED providers. The primary outcome was adherence to clinical practice guidelines, defined as the selection of an appropriate antibiotic agent, dose, and duration of therapy for each patient discharged., Results: When compared to patients discharged from the ED prior to algorithm implementation (N = 325), the post-implementation group (N = 353) received more antibiotic prescriptions that were completely guideline adherent (61.5% vs 11.7%, P < .00001). Post-implementation discharge orders demonstrated improvement in the selection of an appropriate agent (87.3% vs 45.5%, P < .00001), dose (91.5% vs 77.2%, P < .00001), and duration of therapy (71.1% vs 39.1%, P < .01). Additionally, fluoroquinolone prescribing rates were reduced (2.3% vs 12.3%, P < .00001). A reduction in all-cause 30-day returns to the ED or urgent care was observed (15.3% vs 21.5%, P = .036)., Conclusion: Pharmacist-driven implementation of antibiotic prescribing algorithms improved guideline adherence in the outpatient treatment of CAP, SSTI, and UTI.

There is an increasing demand to develop and characterize high moisture extrudates from alternative plant proteins due to their increased use in various foods. In this study, wet texturized proteins from two pea isolates and four oilseed flours from pumpkin and sunflower were subjected to an acid titration to gain insights into their buffering capacity. Results were compared to pork meat with a special emphasis on compositional differences. Wet texturized pumpkin and sunflower proteins had the highest buffering capacity, especially in between pH7.0 and pH4.5, while pea protein extrudates and pork meat were more prone to acidification and similar in buffering capacity. A multiple linear regression model further revealed that ash and select minerals and amino acids are key influencing factors on the overall buffering capacity, while the effect of protein and non-protein nitrogen depends on the evaluated pH-regime. The obtained results underline the importance for a more in-depth physicochemical characterization of texturized plant proteins and their raw materials and suggest a need for recipe and process adjustment to achieve stable pH values., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Patients with Down syndrome and acute lymphocytic leukaemia are at an increased risk of treatment-related mortality and relapse, which is influenced by unfavourable genetic aberrations (eg, IKZF1 deletion). We aimed to investigate the potential underlying effect of Down syndrome versus the effects of adverse cancer genetics on clinical outcome., Method: Patients (aged 1-23 years) with Down syndrome and acute lymphocytic leukaemia and matched non-Down syndrome patients with acute lymphocytic leukaemia (matched controls) from eight trials (DCOG ALL10 and ALL11, ANZCHOG ALL8, AIEOP-BFM ALL2009, UKALL2003, NOPHO ALL2008, CoALL 07-03, and CoALL 08-09) done between 2002 and 2018 across various countries (the Netherlands, the UK, Australia, Denmark, Finland, Iceland, Norway, Sweden, and Germany) were included. Participants were matched (1:3) for clinical risk factors and genetics, including IKZF1 deletion. The primary endpoint was the comparison of MRD levels (absolute MRD levels were categorised into two groups, low [<0·0001] and high [≥0·0001]) between patients with Down syndrome and acute lymphocytic leukaemia and matched controls, and the secondary outcomes were comparison of long-term outcomes (event-free survival, overall survival, relapse, and treatment-related mortality [TRM]) between patients with Down syndrome and acute lymphocytic leukaemia and matched controls. Two matched cohorts were formed: for MRD analyses and for long-term outcome analyses. For both cohorts, matching was based on induction regimen; for the long-term outcome cohort, matching also included MRD-guided treatment group. We used mixed-effect models, Cox models, and competing risk for statistical analyses., Findings: Of 251 children and adolescents with Down syndrome and acute lymphocytic leukaemia, 136 were eligible for analyses and matched to 407 (of 8426) non-Down syndrome patients with acute lymphocytic leukaemia (matched controls). 113 patients with Down syndrome and acute lymphocytic leukaemia were excluded from matching in accordance with predefined rules, no match was available for two patients with Down syndrome and acute lymphocytic leukaemia. The proportion of patients with high MRD at the end of induction treatment was similar for patients with Down syndrome and acute lymphocytic leukaemia (52 [38%] of 136) and matched controls (157 [39%] of 403; OR 0·97 [95% CI 0·64-1·46]; p=0·88). Patients with Down syndrome and acute lymphocytic leukaemia had a higher relapse risk than did matched controls in the IKZF1 deleted group (relapse at 5 years 37·1% [17·1-57·2] vs 13·2% [6·1-23·1]; cause-specific hazard ratio [HR cs ] 4·3 [1·6-11·0]; p=0·0028), but not in the IKZF1 wild-type group (relapse at 5 years 5·8% [2·1-12·2] vs 8·1% [5·1-12·0]; HR cs 1·0 [0·5-2·1]; p=0·99). In addition to increased induction deaths (15 [6%] of 251 vs 69 [0·8%] of 8426), Down syndrome and acute lymphocytic leukaemia was associated with a higher risk of post-induction TRM compared with matched controls (TRM at 5 years 12·2% [7·0-18·9] vs 2·7% [1·3-4·9]; HR cs 5·0 [2·3-10·8]; p<0·0001)., Interpretation: Induction treatment is equivalently effective for patients with Down syndrome and acute lymphocytic leukaemia and for matched patients without Down syndrome. Down syndrome itself provides an additional risk in individuals with IKZF1 deletions, suggesting an interplay between the germline environment and this poor risk somatic aberration. Different treatment strategies are warranted considering both inherent risk of relapse and high risk of TRM., Funding: Stichting Kinder Oncologisch Centrum Rotterdam and the Princess Máxima Center Foundation, NHMRC Australia, The Cancer Council NSW, Tour de Cure, Blood Cancer UK, UK Medical Research Council, Children with Cancer, Swedish Society for Pediatric Cancer, Swedish Childhood Cancer Fund, Danish Cancer Society and the Danish Childhood Cancer Foundation., Competing Interests: Declaration of interests KS reports speaker or advisory board honoraria from Jazz Pharmaceuticals and Servier; speaker fees from Amgen and Medscape; and an educational grant from Servier. CMZ reports grants from Pfizer, Takeda, AbbVie, and Jazz Pharmaceuticals; consulting fees from Novartis, Incyte, Pfizer, Jazz Pharmaceuticals, Takeda, and Abbvie; speaker fees from Pfizer; travel expenses from Jazz Pharmaceuticals; participation on data safety monitoring committees or advisory boards for Novartis, and Incyte; and is co-chair of the Innovative Therapies for Children with Cancer heamatological malignancies committee. GB reports grants from Swedish Society Pediatric Cancer. TT reports foundation funding to Children's Cancer Institute; project funding from Tour de Cure; and ownership of stock or stock options in CSL, Cochlear, Medical Developments International, Osteopore, and Sonic Healthcare. RS reports grants paid to the University of New South Wales from National Health and Medical Research Council Australia, Cancer Counsel New South Wales, and Cancer Australia; and foundation funding to the Children's Cancer Institute from Tour de Cure and Australian Cancer Research Foundation. All other authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

Human neurons engineered from induced pluripotent stem cells (iPSCs) through neurogenin 2 (NGN2) overexpression are widely used to study neuronal differentiation mechanisms and to model neurological diseases. However, the differentiation paths and heterogeneity of emerged neurons have not been fully explored. Here, we used single-cell transcriptomics to dissect the cell states that emerge during NGN2 overexpression across a time course from pluripotency to neuron functional maturation. We find a substantial molecular heterogeneity in the neuron types generated, with at least two populations that express genes associated with neurons of the peripheral nervous system. Neuron heterogeneity is observed across multiple iPSC clones and lines from different individuals. We find that neuron fate acquisition is sensitive to NGN2 expression level and the duration of NGN2-forced expression. Our data reveal that NGN2 dosage can regulate neuron fate acquisition, and that NGN2-iN heterogeneity can confound results that are sensitive to neuron type., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Natural killer (NK) cells play a key role in the control of cancer development, progression and metastatic dissemination. However, tumor cells develop an array of strategies capable of impairing the activation and function of the immune system, including NK cells. In this context, a major event is represented by the establishment of an immunosuppressive tumor microenvironment (TME) composed of stromal cells, myeloid-derived suppressor cells, tumor-associated macrophages, regulatory T cells and cancer cells themselves. The different immunoregulatory cells infiltrating the TME, through the release of several immunosuppressive molecules or by cell-to-cell interactions, cause an impairment of the recruitment of NK cells and other lymphocytes with effector functions. The different mechanisms by which stromal and tumor cells impair NK cell function have been particularly explored in adult solid tumors and, in less depth, investigated and discussed in a pediatric setting. In this review, we will compare pediatric and adult solid malignancies concerning the respective mechanisms of NK cell inhibition, highlighting novel key data in neuroblastoma and Wilms' tumor, two of the most frequent pediatric extracranial solid tumors. Indeed, both tumors are characterized by the presence of stromal cells acting through the release of immunosuppressive molecules. In addition, specific tumor cell subsets inhibit NK cell cytotoxic function by cell-to-cell contact mechanisms likely controlled by the transcriptional coactivator TAZ. These findings could lead to a more performant diagnostic approach and to the development of novel immunotherapeutic strategies targeting the identified cellular and molecular targets.

We generated induced excitatory neurons (iNeurons, iNs) from chimpanzee, bonobo, and human stem cells by expressing the transcription factor neurogenin-2 (NGN2). Single-cell RNA sequencing showed that genes involved in dendrite and synapse development are expressed earlier during iNs maturation in the chimpanzee and bonobo than the human cells. In accordance, during the first 2 weeks of differentiation, chimpanzee and bonobo iNs showed repetitive action potentials and more spontaneous excitatory activity than human iNs, and extended neurites of higher total length. However, the axons of human iNs were slightly longer at 5 weeks of differentiation. The timing of the establishment of neuronal polarity did not differ between the species. Chimpanzee, bonobo, and human neurites eventually reached the same level of structural complexity. Thus, human iNs develop slower than chimpanzee and bonobo iNs, and this difference in timing likely depends on functions downstream of NGN2., Competing Interests: MS, XJ, LF, SE, AW, SK, TS, NN, BT, BP, WH, ET No competing interests declared, (© 2021, Schörnig et al.)

In this study, we examined differential effects of facets of the home language and literacy environment that are known to be relevant to either language development (i.e., quantity and quality of language and literacy stimulation at home) or theory of mind (ToM) development (i.e., parental mental state language), on both children's language skills and their ToM understanding. Moreover, we investigated whether these relations are particularly relevant for children from homes with low socioeconomic status (SES) and whether they account for SES-related disparities in child language skills and ToM understanding. Using longitudinal data of a sample of 224 monolingual German preschool children (assessment of language skills at age 4;6 and 5;6 and ToM at age 5;6), we analyzed the effects of three facets of the home language and literacy environment on later child language and ToM understanding. These facets were book exposure as a measure for quantity of language and literacy stimulation at home, quality of verbal interaction, and parental mental state language assessed between ages 3 and 4. Path analyses showed that book exposure is related to both later ToM understanding and language skills at age 5;6 years; yet, this effect is mediated by earlier language skills at age 4;6 years. Furthermore, book exposure partly mediated the association between SES and language skills and, via earlier language skills at age 4;6, also the relation between SES and ToM. When focusing on children from lower SES families, book exposure and quality of verbal stimulation predicted children's later language skills at age 4;6. Book exposure also predicted change in language skills between age 4;6 and age 5;6. Further, book exposure proved to be significantly associated with children's ToM understanding at age 5;6 via the relation with language skills at 4;6 years. In addition, parental mental state language predicted children's ToM understanding at age 5;6 years. Our findings provide new evidence on how different facets of the home language and literacy environment are related to ToM and language development and their interrelation as well as their SES-related disparities., (Copyright © 2020 Ebert, Lehrl and Weinert.)

It is a major challenge to integrate single-cell sequencing data across experiments, conditions, batches, time points, and other technical considerations. New computational methods are required that can integrate samples while simultaneously preserving biological information. Here, we propose an unsupervised reference-free data representation, cluster similarity spectrum (CSS), where each cell is represented by its similarities to clusters independently identified across samples. We show that CSS can be used to assess cellular heterogeneity and enable reconstruction of differentiation trajectories from cerebral organoid and other single-cell transcriptomic data, and to integrate data across experimental conditions and human individuals.

Purpose: Redundant nerve roots (RNRs) are a negative prognostic factor in patients with central lumbar spinal stenosis (LSS). Forty percent of candidates for surgical decompression show RNRs (RNR+) on preoperative conventional magnetic resonance imaging (MRI). We investigated the prevalence of RNRs in three functional postures (standing, neutral sitting and flexed sitting) with an upright MRI (upMRI)., Methods: A retrospective observational study with a repeated measures design. Thirty surgical candidates underwent upMRI. Sagittal and axial T2-weighted images of the three functional postures were evaluated. The segmental length of the lumbar spine (sLLS), the lordotic angle (LA) and the dural cross-sectional area (DCSA) were measured in each body position. Generalized linear mixed models were carried out. The 0.05 level of probability was set as the criterion for statistical significance., Results: The prevalence of RNRs decreased from 80% during standing to 16.7% during flexed sitting (p < 0.001). The sLLS increased significantly from standing to neutral sitting in both RNR groups (p < 0.001). The increase from neutral sitting to flexed sitting was only significant (p < 0.001) for the group without RNRs (RNR-). The LA decreased significantly for both RNR groups from standing to flexed sitting (p < 0.001). The DSCA increased significantly in the RNR- group (p < 0.001) but not in the RNR+ group (p = 0.9)., Conclusion: The prevalence of RNRs is body position dependent. Increases in DCSA play a determinant role in resolving RNRs.

The present study tests a section of the DIET (direct and indirect effects model of text comprehension; Kim, 2017) model and focuses on the relations between early language skills, various facets of mental state understanding, and text comprehension. In a sample of 267 children, I analyzed the relations between language skills (vocabulary, sentence comprehension) at age 3;6, theory of mind (ToM) at age 5;6, mental state language and metacognitive knowledge at age 9;2, and children's listening and reading comprehension of texts at age 13;7 years. For reading comprehension, results favored a total mediation model that included only direct links from metacognitive knowledge and mental state language to reading comprehension. For listening comprehension, by contrast, a model that also included direct relations from language and ToM in preschool was favored. Metacognitive skills did not mediate the relation between early skills and later text comprehension but, along with mental state language, showed direct relations with reading comprehension beyond listening comprehension. Early language skills showed various indirect relations with later reading comprehension via ToM, mental state language, and listening comprehension, whereas ToM showed only small indirect relations with later reading comprehension via later listening comprehension. These different relations of the various components with later listening in contrast to reading comprehension are discussed., (Copyright © 2020 Ebert.)

This study longitudinally investigated the relation between theory of mind (ToM) and verbal language skills in 231 children from preschool to early adolescence. Further, links to reading comprehension of texts at age 13;7 (years;months) were examined. To assess ToM, children completed false belief tasks at 5;6 and the Strange Stories at 12;8. To assess language, children completed a receptive grammar/sentence comprehension test at 3;6 and 5;6, a receptive vocabulary test at 3;6, 5;6 and 12;8, as well as a test of listening comprehension of texts at 13;7. A bidirectional relation between early and advanced measures of children's language skills and ToM was found: Changes in ToM were predicted by language skills, especially by receptive grammar/sentence comprehension; changes in children's receptive vocabulary were predicted by early ToM. However, early ToM had no direct or indirect effect on later listening comprehension or reading comprehension after controlling for early language skills. Only children's advanced ToM had a small indirect effect on reading comprehension, via listening comprehension. The results are discussed in light of ToM stability over time, and theories on how language and ToM development are intertwined., (Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.)

Belt positioning boosters reduce injury risk for child occupants compared with seat belts alone. While boosters shorten the effective seat length (and thus reduce slouching), "boosting" the child relative to the vehicle interior components also achieves additional safety benefits. First, the increase of the lap belt angle usually improves belt fit across the pelvis and reduces the risk of the occupant slipping ("submarining") under the belt. Second, the torso belt is re-centered over the bony landmarks of the shoulder for more effective/secure restraint. Third, the child's head is relocated in a range better protected by side airbags. The objective of this research was to quantify differences in posture and belt fit across a range of booster designs that provide different levels of boosting. Posture and belt fit were measured in 25 child volunteers aged four to 12. Children were measured in three laboratory seating conditions selected to provide a range of cushion lengths and belt geometries. Six different boosters, as well as a no-booster condition, were evaluated. The low height boosters produced postures that were more slouched, with the hips further forward than in other more typical boosters. Lap belt fit in the low height boosters was not meaningfully different from the other boosters. Shoulder belt fit produced by the lowest height booster was similar to the no-booster condition. Belt positioning boosters that boost the child less than 70 mm produced postures similar to the no-booster condition. While lap belt guides on these products can produce a similar static lap belt fit, they may not provide adequate dynamic performance and do not achieve the other benefits that come with raising the child to a more advantageous location relative to interior components and belts.

Objective: This study presents anthropometric data for individuals with high body mass index (BMI). Modified anthropometric dimensions were also developed to address the challenges of obtaining accurate and repeatable data for this population segment., Background: The prevalence of obesity affects approximately 40% of the U.S. adult population. Anthropometric data are needed to guide product design and safety, but few individuals with high BMI have been measured in available datasets., Method: Anthropometric data for a convenience sample of 288 adults with high BMI (≥30 kg/m 2 ) were collected. To increase participation and minimize participant discomfort, measurements were collected at three bariatric weight loss clinics and one academic research institution., Results: The current obese cohort are heavier than the U.S. general population, with a difference in mean body weight of 47 kg for women and 56 kg for men. The obese cohort are also heavier and have a higher BMI compared with the NIOSH (National Institute for Occupational Safety and Health) truck driver population. Waist circumferential measures of the current obese cohort were larger than women or men in either population compared, a result indicative of meaningful body shape differences., Conclusion: To our knowledge, this study is the first to collect anthropometric data for the obese population segment and conduct comparisons to the U.S. general population and available occupational databases. The obese cohort differed substantially with respect to the distributions of anthropometric variables., Application: These data provide insights about the obese population segment that are relevant to product design, and establish a foundation for future data collection efforts.

Background and Purpose: Neuroinflammation has been proposed as part of the pathogenesis of post-concussion symptoms (PCS), but the inflammatory response of the human brain to mild traumatic brain injury (mTBI) remains unknown. We hypothesized that a neuroinflammatory response is present in mTBI at 1-2 weeks post-injury and persists in patients with PCS., Methods: We scanned 14 patients with mTBI without signs of structural damage at 1-2 weeks and 3-4 months post-injury and 22 healthy controls once using the single photon emission computed tomography tracer 123 I-CLINDE, which visualizes translocator protein (TSPO), a protein upregulated in active immune cells. PCS was defined as three or more persisting symptoms from the Rivermead Post Concussion Symptoms Questionnaire at 3 months post-injury., Results: Across brain regions, patients had significantly higher 123 I-CLINDE binding to TSPO than healthy controls, both at 1-2 weeks after the injury in all patients (P = 0.011) and at 3-4 months in the seven patients with PCS (P = 0.006) and in the six patients with good recovery (P = 0.018). When the nine brain regions were tested separately and results were corrected for multiple comparisons, no individual region differed significantly, but all estimated parameters indicated increased 123 I-CLINDE binding to TSPO, ranging from 2% to 19% in all patients at 1-2 weeks, 13% to 27% in patients with PCS at 3-4 months and -9% to 17% in patients with good recovery at 3-4 months., Conclusions: Neuroinflammation was present in mTBI at 1-2 weeks post-injury and persisted at 3-4 months post-injury with a tendency to be most pronounced in patients with PCS., (© 2019 EAN.)

Unicellular microalgae are a valuable source of macro- and micronutrients. They contain, for example, proteins that are potentially useful as new emulsifiers. The aim of this study was to investigate the emulsifying properties of a less-refined lyophilized crude water-soluble extract (WSE), obtained from the heterotrophically cultivated microalga Chlorella protothecoides. Interfacial tension measurements indicated that mainly the proteins in the extract showed interfacial activity. O/W emulsions were prepared by high-pressure homogenization (1 000 bar, 3 passes) with 5.0 wt % of oil and 2.5 wt % of protein from Chlorella protothecoides, resulting in emulsions having a volume-based mean droplet diameter of d 43 ≤ 1 μm and being stable for at least 7 days. Two different stress tests showed that ( i) protein-stabilized emulsions were resistant to very high salt concentrations (up to 500 mM NaCl), and ( ii) emulsions were stable over a very broad pH range of 2-9, with only minor changes in the particle size d 43 (e.g. with an increase of only 300 nm when the pH was lowered from 5 to 4) compared to whey protein-stabilized emulsions. All WSE emulsions had monomodal particle size distributions and were macro- and microscopically stable during a storage of up to 7 days. The results indicate that the WSE of Chlorella protothecoides has remarkably good emulsifying properties and might be of use as a new emulsifier in various applications in which emulsions are exposed to a broad range of ionic strengths and pH values.

Proton beams used for radiotherapy have potential for superior sparing of normal tissue, although range uncertainties are among the main limiting factors in the accuracy of dose delivery. The aim of this study was to benchmark an N-vinylpyrrolidone based polymer gel to perform three-dimensional measurement of geometric proton beam characteristics and especially to test its suitability as a range probe in combination with an anthropomorphic phantom. For single proton pencil beams as well as for 3×3cm 2 mono-energy layers depth dose profiles, lateral dose distribution at different depths and proton range were evaluated in simple cubic gel phantoms at different energies from 75 to 115MeV and different dose levels. In addition, a 90MeV mono-energetic beam was delivered to an anthropomorphic 3D printed head phantom, which was filled with gel. Subsequently, all phantoms underwent magnetic resonance imaging using an axial pixel size of 0.68-0.98mm and with slice thicknesses of 2 or 3mm to derive a 3-dimensional distribution of the T 2 relaxation time, which correlates with radiation dose. Indices describing lateral dose distribution and proton range were compared against predictions from a treatment planning system (TPS, for cubic and head phantoms) and Monte Carlo simulations (MC, for the head phantom) after manual rigid co-registration with the T 2 relaxation time datasets. For all pencil beams, the FWHM agreement with TPS was better than 1mm or 7%. For the mono-energetic layer, the agreement with TPS in this respect was even better than 0.3mm in each case. With respect to range, results from gel measurements differed no more than 0.9mm (1.6%) from values predicted by TPS. In case of the anthropomorphic phantom, deviations with respect to a nominal range of about 61mm as well as in FWHM were slightly higher, namely within 1.0mm and 1.1mm respectively. Average deviations between gel and TPS/MC were similar (-0.3mm±0.4mm/-0.2±0.5mm). In conclusion, polymer gel dosimetry was found to be a valuable tool to determine geometric proton beam properties three-dimensionally and with high spatial resolution in simple cubic as well as in a more complex anthropomorphic phantom. Post registration range errors of the order of 1mm could be achieved. The additional registration uncertainty (95%) was 1mm., (Copyright © 2018. Published by Elsevier GmbH.)

Targeting of tumor immune escape mechanisms holds enormous therapeutic potential. Still, most patients progress under immune checkpoint blockade and some even become hyperprogressors. To investigate how cancer cells respond to activated but ineffective T cells, we challenged peptide-loaded MCF-7 breast cancer cells with antigen-specific CD8 + T cells in which lytic granules had been destroyed by pretreatment with Concanamycin A. Gene expression analysis after coculture revealed simultaneous induction of PD-L1, IDO1, CEACAM1, and further immunoregulatory checkpoints in breast cancer cells. Strikingly, we further observed gene signatures characteristic for dedifferentiation and acquisition of pluripotency markers including Yamanaka factors. Cognate interaction with nonlytic CD8 + T cells also increased the proportion of stem cell-like cancer cells in a cell-to-cell contact- or (at least) proximity-dependent manner in various cell lines and in primary breast cancer cell cultures; this induction of stem cell-like properties was confirmed by enhanced tumor-forming capacity in immunodeficient mice. Resulting tumors were characterized by enhanced cell density, higher proliferation rates, and increased propensity for lymphoid metastasis. These findings describe a widely underappreciated pathway for immune escape, namely immune-mediated dedifferentiation of breast cancer cells, which is associated with profound changes in gene expression and cellular behavior. As the enhanced malignant potential of cancer cells after nonlytic cognate interactions with CD8 + T cells enables increased tumor growth and metastasis in BALB/c nu/nu mice, the described mechanism may provide a possible explanation for the clinical phenomenon of hyperprogression in response to unsuccessful immunotherapy. SIGNIFICANCE: This study shows that ineffective immune responses not only fail to clear a malignancy, but can also activate pathways in cancer cells that promote stemness and tumor-seeding capacity., (©2019 American Association for Cancer Research.)