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Inherited peripheral neuropathies (IPNs) are a group of diseases associated with mutations in various genes with fundamental roles in the development and function of peripheral nerves. Over the past 10 years, significant advances in identifying molecular disease mechanisms underlying axonal and myelin degeneration, acquired from cellular biology studies and transgenic fly and rodent models, have facilitated the development of promising treatment strategies. However, no clinical treatment has emerged to date. This lack of treatment highlights the urgent need for more biologically and clinically relevant models recapitulating IPNs. For both neurodevelopmental and neurodegenerative diseases, patient-specific induced pluripotent stem cells (iPSCs) are a particularly powerful platform for disease modeling and preclinical studies. In this review, we provide an update on different in vitro human cellular IPN models, including traditional two-dimensional monoculture iPSC derivatives, and recent advances in more complex human iPSC-based systems using microfluidic chips, organoids, and assembloids.

A frequently debated topic related to the origin of life centers around the question of how complex forms of life on today's Earth may have evolved over time from simpler predecessors. For example, the question of how proton concentration gradients across cellular membranes developed in ancestral protocells remains unanswered. This process, which is indispensable for the generation of chemical energy in modern organisms, is driven by energy derived from redox processes in the respiratory chain. Since it is highly unlikely that the complex machinery of the respiratory chain was available on early Earth, we provide an example of how proton gradients can be established in less complex systems. Utilizing liposomes as models of primitive cells, we were able to generate proton gradients of about two pH units across the liposome bilayers using redox reactions as the driving force. Electrons were transferred from sodium sulfite present on the outside of the liposomes to ferricyanide, which was trapped on the inside. A lipid-soluble phenazine derivative served as a shuttle that transferred both electrons and protons across the lipid bilayer. Because sulfite would have been an abundant reduced solute available to the earliest cells, we propose that it may have been a primary source of redox energy for primitive chemiosmotic energy transduction.

Soft tissue sarcomas (STS) are heterogenous malignancies with an unmet need for novel immunotherapies. Tumor infiltrating lymphocytes (TILs) have previously been linked with favorable outcomes in STS patients, though the contribution of natural killer (NK) cell subsets, including NKp46 and CD56bright/dim, has yet to be investigated in detail. Despite the known role of MHC-I on immunoregulation of NK and T cells, limited data exist characterizing the spatial relationship of NK cells to MHC-I+/- cells and T cells in the STS tumor microenvironment (TME). Using STS specimens from 130 patients, we evaluated intratumoral NK cell subsets by immunohistochemistry (IHC), flow cytometry, and immunofluorescence (IF) to assess their impact on overall survival (OS) and metastasis-free survival (MFS). We also assessed the spatial localization of NK and T cells by multiplex IF in the TME, specifically analyzing the effects of MHC-I expression status on NK and T cell clustering. High intratumoral NKp46 expression was associated with improved OS by IHC (P=0.04) and IF (P=0.02). CD56dim NK cells were associated with a survival benefit (P=0.05), while higher infiltrates of CD56bright NK cells predicted worse prognosis (P=0.05). CD3-CD56+ NK cells demonstrated a significant inverse relationship with CD3+ T cells by both flow cytometry and IF. Spatial analyses showed NK cells preferentially clustering close to other NK cells with sparse CD3+ T and CD8+ T cells in range (P

BackgroundNatural killer (NK) cells are cytotoxic cells capable of recognizing heterogeneous cancer targets without prior sensitization, making them promising prospects for use in cellular immunotherapy. Companion dogs develop spontaneous cancers in the context of an intact immune system, representing a valid cancer immunotherapy model. Previously, CD5 depletion of peripheral blood mononuclear cells (PBMCs) was used in dogs to isolate a CD5dim-expressing NK subset prior to co-culture with an irradiated feeder line, but this can limit the yield of the final NK product. This study aimed to assess NK activation, expansion, and preliminary clinical activity in first-in-dog clinical trials using a novel system with unmanipulated PBMCs to generate our NK cell product.MethodsStarting populations of CD5-depleted cells and PBMCs from healthy beagle donors were co-cultured for 14 days, phenotype, cytotoxicity, and cytokine secretion were measured, and samples were sequenced using the 3'-Tag-RNA-Seq protocol. Co-cultured human PBMCs and NK-isolated cells were also sequenced for comparative analysis. In addition, two first-in-dog clinical trials were performed in dogs with melanoma and osteosarcoma using autologous and allogeneic NK cells, respectively, to establish safety and proof-of-concept of this manufacturing approach.ResultsCalculated cell counts, viability, killing, and cytokine secretion were equivalent or higher in expanded NK cells from canine PBMCs versus CD5-depleted cells, and immune phenotyping confirmed a CD3-NKp46+ product from PBMC-expanded cells at day 14. Transcriptomic analysis of expanded cell populations confirmed upregulation of NK activation genes and related pathways, and human NK cells using well-characterized NK markers closely mirrored canine gene expression patterns. Autologous and allogeneic PBMC-derived NK cells were successfully expanded for use in first-in-dog clinical trials, resulting in no serious adverse events and preliminary efficacy data. RNA sequencing of PBMCs from dogs receiving allogeneic NK transfer showed patient-unique gene signatures with NK gene expression trends in response to treatment.ConclusionsOverall, the use of unmanipulated PBMCs appears safe and potentially effective for canine NK immunotherapy with equivalent to superior results to CD5 depletion in NK expansion, activation, and cytotoxicity. Our preclinical and clinical data support further evaluation of this technique as a novel platform for optimizing NK immunotherapy in dogs.

Abstract:

The cancer stem cell (CSC) hypothesis postulates that heterogeneous human cancers harbor a population of stem-like cells which are resistant to cytotoxic therapies, thus providing a reservoir of relapse following conventional therapies like chemotherapy and radiation (RT). CSCs have been observed in multiple human cancers, and their presence has been correlated with worse clinical outcomes. Here, we sought to evaluate the impact of drug dosing of the multi-tyrosine kinase inhibitor, sorafenib, on CSC and non-CSCs in soft tissue sarcoma (STS) models, hypothesizing differential effects of sorafenib based on dose and target cell population. In vitro, human cancer cell lines and primary STS from surgical specimens were exposed to escalating doses of sorafenib to determine cell viability and expression of CSC marker aldehyde dehydrogenase (ALDH). In vivo, ALDHbright CSCs were isolated, exposed to sorafenib, and xenograft growth and survival analyses were performed. We observed that sarcoma CSCs appear to paradoxically respond to the tyrosine kinase inhibitor sorafenib at low doses with increased proliferation and stem-like function of CSCs, whereas anti-viability effects dominated at higher doses. Importantly, STS patients receiving neoadjuvant sorafenib and RT on a clinical trial (NCT00864032) showed increased CSCs post therapy, and higher ALDH scores post therapy were associated with worse metastasis-free survival. These data suggest that low-dose sorafenib may promote the CSC phenotype in STS with clinically significant effects, including increased tumor growth and higher rates of metastasis formation in sarcoma patients.

INTRODUCTION: The global human population is still growing such that our collective enterprise is driving environmental catastrophe. Despite a decline in average population growth rate, we are still experiencing the highest annual increase of global human population size in the history of our species-averaging an additional 84 million people per year since 1990. No review to date has accumulated the available evidence describing the associations between increasing population and environmental decline, nor solutions for mitigating the problems arising. METHODS: We summarize the available evidence of the relationships between human population size and growth and environmental integrity, human prosperity and wellbeing, and climate change. We used PubMed, Google Scholar, and Web of Science to identify all relevant peer-reviewed and gray-literature sources examining the consequences of human population size and growth on the biosphere. We reviewed papers describing and quantifying the risks associated with population growth, especially relating to climate change. RESULTS: These risks are global in scale, such as greenhouse-gas emissions, climate disruption, pollution, loss of biodiversity, and spread of disease-all potentially catastrophic for human standards of living, health, and general wellbeing. The trends increasing the risks of global population growth are country development, demographics, maternal education, access to family planning, and child and maternal health. CONCLUSION: Support for nations still going through a demographic transition is required to ensure progress occurs within planetary boundaries and promotes equity and human rights. Ensuring the wellbeing for all under this aim itself will lower population growth and further promote environmental sustainability.

PURPOSE: Emerging evidence suggests that osteosarcoma stem cells (OSCs) may be responsible for tumor initiation propagation, recurrence, and resistance to therapy. We set out to evaluate the relationship between the abundance of ALDH1A1 and CD44-positive cells in biopsy and resection samples on disease recurrence and overall survival. METHODS: A retrospective review of 20 patients, including biopsy and resection samples, was performed at a comprehensive cancer center. Additionally, we queried the publicly available TARGET dataset of osteosarcoma patients. RESULTS: Neither the percentages of ALDH1A1-positive cells nor CD44-positive cells were significantly associated with overall mortality or disease recurrence in either biopsy or resection samples. Unlike our institutional data, overall survival was significantly correlated to higher ALDH1A1 expression in the TARGET dataset both in univariate and age-adjusted analyses. CONCLUSIONS: ADLH1 and CD44, potential markers of OSCs, were not found to be reliable clinical immunohistochemical prognostic markers for osteosarcoma patient survival, specifically disease-free survival. Osteosarcoma patients with high ALDH1A1 RNA expression showed improved overall survival in examining a national genomic database of osteosarcoma patients but again no association with disease-free survival. The potential of CD44 and ALDH1A1 as cellular-specific prognostic markers of survival, and as possible molecular targets, may be limited in osteosarcoma.

Abstract Background Natural HbA1c levels in GCK Maturity-onset diabetes of the young (GCK-MODY) patients often sit above the diagnostic threshold for type 2 diabetes (T2D). Treatments to lower HbA1c levels show reduced effectiveness in these individuals, yet in case studies to date, GCK-MODY patients often evade secondary T2D complications. Given these deviations, genetic screening of GCK may be clinically useful, but population studies are needed to more broadly understand T2D-related complications in GCK variant carriers. Methods To identify GCK variant carriers at the population level, we used both ACMG/AMP variant interpretation for GCK-MODY pathogenicity and a state-of-the-art variant interpretation strategy based on functional and statistical evidence to predict glucose elevations. Presence of pathogenic and glucose-elevating GCK variants was assessed in two cohorts (n~535,000). We identified 442 individuals with GCK variants predicted to increase glucose (~1/1200), with 150 (34%) of these individuals harboring variants reaching a pathogenic interpretation. Results In a retrospective analysis, we show that in addition to elevated HbA1c, pathogenic variant carriers are 10x as likely, and all other glucose-elevating GCK variant carriers are 3x as likely, to receive a T2D diagnosis compared to non-GCK carriers. Surprisingly, carriers of pathogenic and glucose-elevating GCK variants with T2D develop T2D-related complications at rates more than double that of individuals without T2D, comparable to non-GCK individuals with T2D. Conclusions This population-level assessment shows secondary complications in individuals with pathogenic and glucose-elevating GCK variants and T2D and suggests that genotyping for these variants should be considered in a precision medicine approach for T2D treatment and prevention.

Abstract The modular multilevel DC–DC transformer (MMDCT) provides a reliable solution to overcome the challenges of conventional dual‐active‐bridge converters in terms of power semiconductors ratings and dv/dt stress on transformer coupling windings. An alternative modulation method, quasi‐square‐wave, was proposed to reduce the cell capacitance of modular multilevel bridges. However the application of quasi‐square‐wave modulation is found to result in underdamped switching transients and losses when resetting energy stored in arm inductance. This paper presents a detailed transient analysis of MMDCT arm insertion based on an equivalent circuit, which contributes to a more accurate component sizing and gives voltage estimation for individual half‐bridge submodules. Additionally, a revised switching sequence is proposed to recover this inductor energy and lower the oscillation‐related losses. Simulated and experimental results from a scaled test rig of MMDCT are implemented and validate the proposed component sizing and switching sequence, indicating that the converter efficiency can be improved under revised switching sequence. Finally, a silicon carbide based, high‐frequency MMDCT is proposed and simulated.

Abstract Background Preoperative health optimisation for elective surgery entails supporting patients to improve their health in preparation for their treatment and recovery. While there is consensus that this process should address obesity, approaches vary across England. Despite guidance from the National Institute for Health and Care Excellence to the contrary, restrictive approaches with body mass index thresholds for referral to arthroplasty are in use. This qualitative study aimed to investigate the views of professionals on the current use and future implications of these policies. Methods Semi-structured interviews were conducted with 20 professionals including clinicians, commissioners, policymakers, and health service managers, with experience of developing and/or implementing health optimisation policies for elective arthroplasty. Participants were sampled from areas in England with and without restrictive policies. We undertook thematic analysis of the interview data. Results Participants described pre-surgical health optimisation as an important trigger for health improvement but identified current resourcing and inadequacies in provision of weight management support as significant barriers to success. Participants expressed concerns about the appropriateness and fairness of including obesity as a determinant to restrict access to surgery. They described short-term financial pressures underlying the use of restrictive body mass index thresholds and a lack of an evidence base, such that policies amounted to rationing and risked exacerbations of health inequalities. The study identified four priorities for improvements to future health optimisation practices: developing and implementing national guidance with flexibility for local variation, initiating patient engagement in primary care with onward integration across all services, improving resourcing to support effective equitable impact, and addressing wider determinants of obesity through societal change. Conclusions Overall, participants had limited expectations of the impact of health optimisation policies on obesity without additional support, investment, and national guideline implementation. They raised strong concerns over current restrictive approaches. We conclude that addressing concerns around weight management support service availability and impacts on health inequalities is essential for shaping effective health optimisation policies. Future policy direction should support health optimisation to be offered early (ideally in primary care). Health optimisation interventions should be non-restrictive, inclusive, and well-monitored, particularly around equality impact.

Abstract Active gate driving (AGD) is a promising concept for achieving high‐performance power transistor switching. This is particularly crucial for Silicon Carbide (SiC) MOSFETs since their inherently fast switching characteristics give rise to severe overshoots and oscillations which translate into increased levels of electromagnetic interference (EMI) emissions. In this paper, an AGD strategy using a single‐pulse applied during the switching transient is considered for a 1200 V 400 A SiC MOSFET module. The effect of single‐pulse timing, load current, and temperature on the switching performance is analyzed in detail. The radiated EMI reduction benefits are quantified by H‐field and E‐field probes. A conceptual closed‐loop AGD approach is presented and compared to open‐loop operation. For the transistor turn‐off case under full load current of 400 A, experimental results show that it is possible to reduce voltage overshoot by 43.3%, voltage and current oscillations by 69.7% and 52.2% respectively, and EMI by 76.6%, with a trade‐off in the switching energy by a relatively minor increase of 18.2%, compared to the conventional gate driving case. For the turn‐on case, current overshoot was reduced by 32.7%, EMI by 52%, voltage and current oscillations by 54.6% and 52.8%, respectively, with a penalty of 50.9% increase in the switching loss.

Abstract Background Preterm birth is a major cause of infant mortality and morbidity and accounts for 7–8% of births in the UK. It is more common in women from socially deprived areas and from minority ethnic groups, but the reasons for this disparity are poorly understood. To inform interventions to improve child survival and their quality of life, this study examined the socioeconomic and ethnic inequalities in preterm births (

People regularly read multi-line texts in different formats and publishers, internationally, must decide how to present text to make reading most effective and efficient. Relatively few studies have examined multi-line reading, and fewer still Chinese multi-line reading. Here, we examined whether texts presented in single or double columns, and either left-justified or fully-justified affect Chinese reading. Text format had minimal influence on overall reading time; however, it significantly impacted return-sweeps (large saccades moving the eyes from the end of one line of text to the beginning of the next). Return-sweeps were launched and landed further away from margins and involved more corrective saccades in single- than double-column format. For left- compared to fully-justified format, return-sweeps were launched and landed closer to margins. More corrective saccades also occurred. Our results showed more efficient return-sweep behavior for fully- than left-justified text. Moreover, there were clear trade-off effects such that formats requiring increased numbers of shorter return-sweeps produced more accurate targeting and reduced numbers of corrective fixations, whereas formats requiring reduced numbers of longer return-sweeps caused less accurate targeting and an increased rate of corrective fixations. Overall, our results demonstrate that text formats substantially affect return-sweep eye movement behavior during Chinese reading without affecting efficiency and effectiveness, that is, the overall time it takes to read and understand the text.

Background: People who have communication difficulties may benefit from using augmentative and alternative communication (AAC). Understanding and measuring outcomes from the use of AAC is an important part of evaluating the impact of devices and services. Outcome measurement needs to reflect the changing nature of the impact of using AAC on an individual's ability to participate in activities of daily life. There is a limited understanding of the concepts that should inform the evaluation of outcomes from AAC device provision, nor how people's expectations from AAC may change over time. Aims: To inform the development of a patient-reported outcome measure for AAC by understanding more about people's expectations from AAC and how these change over time. Methods & Procedures: A longitudinal qualitative research study was designed and carried out with seven participants over a period of 2 years. Participants were recruited from a regional specialist assessment service for AAC in the south-west of the UK. Four semi-structured interviews were carried out: (1) before assessment for AAC, (2) after assessment, (3) directly after provision of an AAC device and (4) between 6 and 12 months after provision. An original analytic method was used in this study that built on the principles of longitudinal interpretative phenomenology analysis, applied with a dialogic theoretical lens. This approach enabled the inclusion of a range of multimodal and embodied data collected to this study and allowed the research team to draw out salient themes across the cohort group while attending to the influence of time and context on experience. Outcomes & Results: The results confirm and extend the three core concepts that were used to guide analysis: changes; contexts; future possibilities. The contextual and temporal influences on outcomes attainable from AAC for this cohort were also identified and illustrated through cross-case comparison. Deeper, analytic, and conceptual engagement with theory, which was then applied to analysis of the data, provided methodological rigour in the study. The results enhance our understanding of people's hopes and expectations from AAC and how these change over time. Conclusions & Implications: This qualitative longitudinal research study provides new insights into the journeys of people who experience communication disability, and the shifting nature of their sense of identity as they engage with, and learn from using, AAC. The study is significant as it attends to the dynamic nature of experience and how contextual and experiential factors influence people's hopes and expectations from AAC. The paper presents an original application of longitudinal qualitative research methodology with people who use AAC which can be further applied and tested in the field of communication disability research.

Award-bearing transnational professional development training has received little attention in the literature. By taking a longitudinal mixed-methods approach, this project's researchers investigated the ways in which participants on a World Bank-funded programme practised leadership at the start of their training, before revisiting them a year later to find out what, if any, changes had resulted. It was discovered that the award-bearing design had been very influential in endowing the participants with concepts that they enthusiastically adopted, but that, over time, the concepts had undergone a process of simplification, largely driven by incongruities between the concepts and the cultural environments to which they had been applied. It is recommended that award-bearing programmes might more readily take into account the individual and contextual circumstances of their participants at the planning stage.

Bystander activation of memory T cells occurs via cytokine signaling alone in the absence of T cell receptor (TCR) signaling and provides a means of amplifying T cell effector responses in an antigen-nonspecific manner. While the role of Programmed Cell Death Protein 1 (PD-1) on antigen-specific T cell responses is extensively characterized, its role in bystander T cell responses is less clear. We examined the role of the PD-1 pathway during human and mouse non-antigen-specific memory T cell bystander activation and observed that PD-1+ T cells demonstrated less activation and proliferation than activated PD-1- populations in vitro. Higher activation and proliferative responses were also observed in the PD-1- memory population in both mice and patients with cancer receiving high-dose IL-2, mirroring the in vitro phenotypes. This inhibitory effect of PD-1 could be reversed by PD-1 blockade in vivo or observed using memory T cells from PD-1-/- mice. Interestingly, increased activation through abrogation of PD-1 signaling in bystander-activated T cells also resulted in increased apoptosis due to activation-induced cell death (AICD) and eventual T cell loss in vivo. These results demonstrate that the PD-1/PD-Ligand 1 (PD-L1) pathway inhibited bystander-activated memory T cell responses but also protected cells from AICD.

BACKGROUND: Dilated cardiomyopathy (DCM) can lead to advanced disease, defined herein as necessitating a durable left ventricular assist device or a heart transplant (LVAD/HT). DCM is known to have a genetic basis, but the association of rare variant genetics with advanced DCM has not been studied. METHODS: We analyzed clinical and genetic sequence data from patients enrolled between 2016 and 2021 in the US multisite DCM Precision Medicine Study, which was a geographically diverse, multiracial, multiethnic cohort. Clinical evaluation included standardized patient interview and medical record query forms. DCM severity was classified into 3 groups: patients with advanced disease with LVAD/HT; patients with an implantable cardioverter defibrillator (ICD) only; or patients with no ICD or LVAD/HT. Rare variants in 36 DCM genes were classified as pathogenic or likely pathogenic or variants of uncertain significance. Confounding factors we considered included demographic characteristics, lifestyle factors, access to care, DCM duration, and comorbidities. Crude and adjusted associations between DCM severity and rare variant genetic findings were assessed using multinomial models with generalized logit link. RESULTS: Patients mean (SD) age was 51.9 (13.6) years; 42% were of African ancestry, 56% were of European ancestry, and 44% were female. Of 1198 patients, 347 had LVAD/HT, 511 had an ICD, and 340 had no LVAD/HT or ICD. The percentage of patients with pathogenic or likely pathogenic variants was 26.2%, 15.9%, and 15.0% for those with LVAD/HT, ICD only, or neither, respectively. After controlling for sociodemographic characteristics and comorbidities, patients with DCM with LVAD/HT were more likely than those without LVAD/HT or ICD to have DCM-related pathogenic or likely pathogenic rare variants (odds ratio, 2.3 [95% CI, 1.5-3.6]). The association did not differ by ancestry. Rare variant genetic findings were similar between patients with DCM with an ICD and those without LVAD/HT or ICD. CONCLUSIONS: Advanced DCM was associated with higher odds of rare variants in DCM genes adjudicated as pathogenic or likely pathogenic, compared with individuals with less severe DCM. This finding may help assess the risk of outcomes in management of patients with DCM and their at-risk family members. REGISTRATION: URL: https://www. CLINICALTRIALS: gov; Unique identifier: NCT03037632.