Exploring seizure management in hospitals, unmet need, and the impact of the COVID-19 pandemic on seizure presentations to hospital

Purpose This study assesses investigations, referrals and admissions in patients presenting to the Emergency Department (ED) with seizures, and the effect of the COVID-19 pandemic on such management. Outcomes in patients with learning disabilities, active significant mental health concerns, and from the most socioeconomically deprived areas were compared to those of the general cohort. Methods Investigations, referrals and admissions were recorded for 120 patients across two cohorts; pre-pandemic (September 2019) and during the pandemic (December 2020). Retrospective review of individual patient electronic health care records was used for data collection. Results There was a decrease in patient numbers from 2019 to 2020. A greater proportion of patients presented with organic cause seizures and fewer presented with non-epileptic attacks. Frequent use of CT heads (45%) is likely to represent improper use of limited resources. There were low referral rates, both to acute neurology (28%) and to the adult epilepsy team (32%). Patients with active significant mental health concerns were significantly less likely to be referred to neurology or admitted. Conclusions Despite a greater proportion of admissions during the Covid-19 pandemic, referrals to acute neurology and the epilepsy team remained low. Failure to refer prevents the most vulnerable seizure patients from receiving appropriate support, as seen in patients with active significant mental health concerns. Neurology staff were unaware of a significant number of patients presenting with seizures, which is of concern in an already over-stretched department. This offers an opportunity to improve care for people with epilepsy

Defined Culture Environments Create an Improved Human Intestinal Organoid Model System to Study Intestinal Development

Organoids are small, stem cell-derived culture systems that mimic aspects of the structure and function of the organs they are modelled after. Thus, organoids provide a 3D model for studying human development and disease in a complex human-derived in vitro system, and offer advantages over traditionally utilized 2D in vitro cell culture platforms or in vivo animal models. Intestinal organoids have been well characterized and used for over a decade to model intestinal pathologies and advance our understanding of intestinal biology. However, intestinal organoid models have been limited by a reliance on commercial basement membrane extracellular matrix (ECM) products such as Matrigel which introduce experimental variability, limit experimental control, and are unsuitable for downstream clinical applications due to their xenogeneic origin. Additionally, current intestinal organoids are relatively immature and do not contain many of the key cell types found in the human intestine. In particular, a serosal mesothelium, the outermost layer of the intestine that provides a protective boundary for the gut, has not been observed within previous in vitro intestinal cell models. In this dissertation, I describe improved culture methods for pluripotent stem cell-derived human intestinal organoids (HIOs) that eliminate reliance on Matrigel and more faithfully recapitulate the organization of the human small intestine. HIOs contain both epithelial and mesenchymal compartments, which enables the formation of a supportive niche within the organoid. Thus, HIOs can be cultured in bioinert environments, including unmodified alginate hydrogels and even suspension culture. Alginate and suspension culture provide simple, cost-effective culture systems for HIOs that offer increased experimental control and decreased variability compared to Matrigel. My studies demonstrate that alginate and suspension culture are effective replacements for Matrigel to support the HIO epithelium, as HIOs cultured in alginate and suspension give rise to expected intestinal epithelial cell types. Additionally, HIOs cultured in bioinert conditions (alginate or suspension) form an organized outer mesenchymal layer that closely resembles the human intestine. Strikingly, HIOs cultured in alginate and suspension form an outer serosal mesothelium that has not been previously observed in Matrigel HIOs. This serosa formation is enhanced in suspension culture compared to alginate. I characterized HIO-serosa to demonstrate that it is molecularly and functionally similar to human intestinal serosal mesothelium. I then utilized suspension HIOs as a model to investigate serosal development and identified roles for Hedgehog (HH) and WNT signaling in human intestinal serosa formation and patterning. Overall, this work provides improved, defined culture methods for human intestinal organoids that better recapitulate the native human intestine for enhanced studies of intestinal development and disease modelling.PHDBiomedical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/174293/1/mmcapeli_1.pd

Entrepreneurial approach and open innovation. A bibliometric and literature review

Since the first definition of Open Innovation the indivisible relationship between open innovation and entrepreneurship has drawn a lot of research attention, however, the exact mechanisms by which openness benefits entrepreneurs and vice versa are not yet fully understood. Based on this consideration, this study aims to offer an overview of the evolution of the literature regarding the OI-entrepreneurship relationship published over the last decades. We use a bibliometric analysis coupled with a systematic literature review were performed over a data set of 106 peer-reviewed articles published from 2005 to 2020. The authors discovered the existence of five well-polarized clusters with the following thematic focusses: entrepreneurial opportunity for OI, strategic partnership opportunity for OI, organizational opportunity for OI, digital opportunity for OI, and institutional opportunity for OI. The authors investigated each thematic cluster by reviewing the most impactful contributions within them to create a framework which my highlights future avenues for the whole topic

Integrative genome-scale analyses reveal post-transcriptional signatures of early human small intestinal development in a directed differentiation organoid model

Abstract Background MicroRNAs (miRNAs) are important post-transcriptional gene regulators controlling cellular lineage specification and differentiation during embryonic development, including the gastrointestinal system. However, miRNA-mediated regulatory mechanisms involved in early embryonic development of human small intestine (SI) remains underexplored. To explore candidate roles for miRNAs in prenatal SI lineage specification in humans, we used a multi-omic analysis strategy in a directed differentiation model that programs human pluripotent stem cells toward the SI lineage. Results We leveraged small RNA-seq to define the changing miRNA landscape, and integrated chromatin run-on sequencing (ChRO-seq) and RNA-seq to define genes subject to significant post-transcriptional regulation across the different stages of differentiation. Small RNA-seq profiling revealed temporal dynamics of miRNA signatures across different developmental events of the model, including definitive endoderm formation, SI lineage specification and SI regional patterning. Our multi-omic, integrative analyses showed further that the elevation of miR-182 and reduction of miR-375 are key events during SI lineage specification. We demonstrated that loss of miR-182 leads to an increase in the foregut master marker SOX2. We also used single-cell analyses in murine adult intestinal crypts to support a life-long role for miR-375 in the regulation of Zfp36l2. Finally, we uncovered opposing roles of SMAD4 and WNT signaling in regulating miR-375 expression during SI lineage specification. Beyond the mechanisms highlighted in this study, we also present a web-based application for exploration of post-transcriptional regulation and miRNA-mediated control in the context of early human SI development. Conclusion The present study uncovers a novel facet of miRNAs in regulating prenatal SI development. We leveraged multi-omic, systems biology approaches to discover candidate miRNA regulators associated with early SI developmental events in a human organoid model. In this study, we highlighted miRNA-mediated post-transcriptional regulation relevant to the event of SI lineage specification. The candidate miRNA regulators that we identified for the other stages of SI development also warrant detailed characterization in the future

Nonadhesive Alginate Hydrogels Support Growth of Pluripotent Stem Cell-Derived Intestinal Organoids

Summary: Human intestinal organoids (HIOs) represent a powerful system to study human development and are promising candidates for clinical translation as drug-screening tools or engineered tissue. Experimental control and clinical use of HIOs is limited by growth in expensive and poorly defined tumor-cell-derived extracellular matrices, prompting investigation of synthetic ECM-mimetics for HIO culture. Since HIOs possess an inner epithelium and outer mesenchyme, we hypothesized that adhesive cues provided by the matrix may be dispensable for HIO culture. Here, we demonstrate that alginate, a minimally supportive hydrogel with no inherent cell instructive properties, supports HIO growth in vitro and leads to HIO epithelial differentiation that is virtually indistinguishable from Matrigel-grown HIOs. In addition, alginate-grown HIOs mature to a similar degree as Matrigel-grown HIOs when transplanted in vivo, both resembling human fetal intestine. This work demonstrates that purely mechanical support from a simple-to-use and inexpensive hydrogel is sufficient to promote HIO survival and development. : Spence and colleagues identify alginate as a nonadhesive growth matrix to support human intestinal organoid culture. Their results demonstrate that alginate-grown HIOs are virtually indistinguishable from traditional Matrigel-grown HIOs both in vitro and in vivo. This work increases the translational potential of HIOs by eliminating reliance on cell-derived matrices and reducing cost. Keywords: intestine, alginate, hydrogel, human pluripotent stem cell, organoid, intestinal organoid, enteroi

Organoids as tools for fundamental discovery and translation - a Keystone Symposia report

Complex three-dimensional in vitro organ-like models, or organoids, offer a unique biological tool with distinct advantages over two-dimensional cell culture systems, which can be too simplistic, and animal models, which can be too complex and may fail to recapitulate human physiology and pathology. Significant progress has been made in driving stem cells to differentiate into different organoid types, though several challenges remain. For example, many organoid models suffer from high heterogeneity, and it can be difficult to fully incorporate the complexity of in vivo tissue and organ development to faithfully reproduce human biology. Successfully addressing such limitations would increase the viability of organoids as models for drug development and preclinical testing. On April 3-6, 2022, experts in organoid development and biology convened at the Keystone Symposium "Organoids as Tools for Fundamental Discovery and Translation" to discuss recent advances and insights from this relatively new model system into human development and disease

Charting human development using a multi-endodermal organ atlas and organoid models

Organs are composed of diverse cell types that traverse transient states during organogenesis. To interrogate this diversity during human development, we generate a single-cell transcriptome atlas from multiple developing endodermal organs of the respiratory and gastrointestinal tract. We illuminate cell states, transcription factors, and organ-specific epithelial stem cell and mesenchyme interactions across lineages. We implement the atlas as a high-dimensional search space to benchmark human pluripotent stem cell (hPSC)-derived intestinal organoids (HIOs) under multiple culture conditions. We show that HIOs recapitulate reference cell states and use HIOs to reconstruct the molecular dynamics of intestinal epithelium and mesenchyme emergence. We show that the mesenchyme-derived niche cue NRG1 enhances intestinal stem cell maturation in vitro and that the homeobox transcription factor CDX2 is required for regionalization of intestinal epithelium and mesenchyme in humans. This work combines cell atlases and organoid technologies to understand how human organ development is orchestrated.ISSN:0092-8674ISSN:1097-417

Organoids as tools for fundamental discovery and translation-A Keystone Symposia report

Complex three-dimensional in vitro organ-like models, or organoids, offer a unique biological tool with distinct advantages over two-dimensional cell culture systems, which can be too simplistic, and animal models, which can be too complex and may fail to recapitulate human physiology and pathology. Significant progress has been made in driving stem cells to differentiate into different organoid types, though several challenges remain. For example, many organoid models suffer from high heterogeneity, and it can be difficult to fully incorporate the complexity of in vivo tissue and organ development to faithfully reproduce human biology. Successfully addressing such limitations would increase the viability of organoids as models for drug development and preclinical testing. On April 3-6, 2022, experts in organoid development and biology convened at the Keystone Symposium "Organoids as Tools for Fundamental Discovery and Translation" to discuss recent advances and insights from this relatively new model system into human development and disease