RNA sequencing reveals MMP2 and TGFB1 downregulation in LRRK2 G2019S Parkinson's iPSC-derived astrocytes

Non-neuronal cell types such as astrocytes can contribute to Parkinson's disease (PD) pathology. The G2019S mutation in leucine-rich repeat kinase 2 (LRRK2) is one of the most common known causes of familial PD. To characterize its effect on astrocytes, we developed a protocol to produce midbrain-patterned astrocytes from human induced pluripotent stem cells (iPSCs) derived from PD LRRK2 G2019S patients and healthy controls. RNA sequencing analysis revealed the downregulation of genes involved in the extracellular matrix in PD cases. In particular, transforming growth factor beta 1 (TGFB1), which has been shown to inhibit microglial inflammatory response in a rat model of PD, and matrix metallopeptidase 2 (MMP2), which has been shown to degrade α-synuclein aggregates, were found to be down-regulated in LRRK2 G2019S astrocytes. Our findings suggest that midbrain astrocytes carrying the LRRK2 G2019S mutation may have reduced neuroprotective capacity and may contribute to the development of PD pathology

Assessing similarity to primary tissue and cortical layer identity in induced pluripotent stem cell-derived cortical neurons through single-cell transcriptomics

Induced pluripotent stem cell (iPSC)-derived cortical neurons potentially present a powerful new model to understand corticogenesis and neurological disease. Previous work has established that differentiation protocols can produce cortical neurons, but little has been done to characterize these at cellular resolution. In particular, it is unclear to what extent in vitro two-dimensional, relatively disordered culture conditions recapitulate the development of in vivo cortical layer identity. Single-cell multiplex reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) was used to interrogate the expression of genes previously implicated in cortical layer or phenotypic identity in individual cells. Totally, 93.6% of single cells derived from iPSCs expressed genes indicative of neuronal identity. High proportions of single neurons derived from iPSCs expressed glutamatergic receptors and synaptic genes. And, 68.4% of iPSC-derived neurons expressing at least one layer marker could be assigned to a laminar identity using canonical cortical layer marker genes. We compared single-cell RNA-seq of our iPSC-derived neurons to available single-cell RNA-seq data from human fetal and adult brain and found that iPSC-derived cortical neurons closely resembled primary fetal brain cells. Unexpectedly, a subpopulation of iPSC-derived neurons co-expressed canonical fetal deep and upper cortical layer markers. However, this appeared to be concordant with data from primary cells. Our results therefore provide reassurance that iPSC-derived cortical neurons are highly similar to primary cortical neurons at the level of single cells but suggest that current layer markers, although effective, may not be able to disambiguate cortical layer identity in all cells

Single-cell sequencing of iPSC-Dopamine neurons reconstructs disease progression and identifies HDAC4 as a regulator of Parkinson cell phenotypes

Induced pluripotent stem cell (iPSC)-derived dopamine neurons provide an opportunity to model Parkinson’s disease (PD), but neuronal cultures are confounded by asynchronous and heterogeneous appearance of disease phenotypes in vitro. Using high-resolution, single-cell transcriptomic analyses of iPSC-derived dopamine neurons carrying the GBA-N370S PD risk variant, we identified a progressive axis of gene expression variation leading to endoplasmic reticulum stress. Pseudotime analysis of genes differentially expressed (DE) along this axis identified the transcriptional repressor histone deacetylase 4 (HDAC4) as an upstream regulator of disease progression. HDAC4 was mislocalized to the nucleus in PD iPSC-derived dopamine neurons and repressed genes early in the disease axis, leading to late deficits in protein homeostasis. Treatment of iPSC-derived dopamine neurons with HDAC4-modulating compounds upregulated genes early in the DE axis and corrected PD-related cellular phenotypes. Our study demonstrates how single-cell transcriptomics can exploit cellular heterogeneity to reveal disease mechanisms and identify therapeutic targets

Targeted health and social care interventions for women and infants who are disproportionately impacted by health inequalities in high-income countries: a systematic review

Background: Disadvantaged populations (such as women from minority ethnic groups and those with social complexity) are at an increased risk of poor outcomes and experiences. Inequalities in health outcomes include preterm birth, maternal and perinatal morbidity and mortality, and poor-quality care. The impact of interventions is unclear for this population, in high-income countries (HIC). The review aimed to identify and evaluate the current evidence related to targeted health and social care service interventions in HICs which can improve health inequalities experienced by childbearing women and infants at disproportionate risk of poor outcomes and experiences. Methods: Twelve databases searched for studies across all HICs, from any methodological design. The search concluded on 8/11/22. The inclusion criteria included interventions that targeted disadvantaged populations which provided a component of clinical care that differed from standard maternity care. Results: Forty six index studies were included. Countries included Australia, Canada, Chile, Hong Kong, UK and USA. A narrative synthesis was undertaken, and results showed three intervention types: midwifery models of care, interdisciplinary care, and community-centred services. These intervention types have been delivered singularly but also in combination of each other demonstrating overlapping features. Overall, results show interventions had positive associations with primary (maternal, perinatal, and infant mortality) and secondary outcomes (experiences and satisfaction, antenatal care coverage, access to care, quality of care, mode of delivery, analgesia use in labour, preterm birth, low birth weight, breastfeeding, family planning, immunisations) however significance and impact vary. Midwifery models of care took an interpersonal and holistic approach as they focused on continuity of carer, home visiting, culturally and linguistically appropriate care and accessibility. Interdisciplinary care took a structural approach, to coordinate care for women requiring multi-agency health and social services. Community-centred services took a place-based approach with interventions that suited the need of its community and their norms. Conclusion: Targeted interventions exist in HICs, but these vary according to the context and infrastructure of standard maternity care. Multi-interventional approaches could enhance a targeted approach for at risk populations, in particular combining midwifery models of care with community-centred approaches, to enhance accessibility, earlier engagement, and increased attendance. Trial registration: PROSPERO Registration number: CRD42020218357

A multi-center prospective study of plant-based nutritional support in adult community-based patients at risk of disease-related malnutrition

IntroductionThere is an emerging need for plant-based, vegan options for patients requiring nutritional support.MethodsTwenty-four adults at risk of malnutrition (age: 59 years (SD 18); Sex: 18 female, 6 male; BMI: 19.0 kg/m2 (SD 3.3); multiple diagnoses) requiring plant-based nutritional support participated in a multi-center, prospective study of a (vegan suitable) multi-nutrient, ready-to-drink, oral nutritional supplement (ONS) [1.5 kcal/mL; 300 kcal, 12 g protein/200 mL bottle, mean prescription 275 mL/day (SD 115)] alongside dietary advice for 28 days. Compliance, anthropometry, malnutrition risk, dietary intake, appetite, acceptability, gastrointestinal (GI) tolerance, nutritional goal(s), and safety were assessed.ResultsPatients required a plant-based ONS due to personal preference/variety (33%), religious/cultural reasons (28%), veganism/reduce animal-derived consumption (17%), environmental/sustainability reasons (17%), and health reasons (5%). Compliance was 94% (SD 16). High risk of malnutrition (‘MUST’ score ≥ 2) reduced from 20 to 16 patients (p = 0.046). Body weight (+0.6 kg (SD 1.2), p = 0.02), BMI (+0.2 kg/m2 (SD 0.5), p = 0.03), total mean energy (+387 kcal/day (SD 416), p < 0.0001) and protein intake (+14 g/day (SD 39), p = 0.03), and the number of micronutrients meeting the UK reference nutrient intake (RNI) (7 vs. 14, p = 0.008) significantly increased. Appetite (Simplified Nutritional Appetite Questionnaire (SNAQ) score; p = 0.13) was maintained. Most GI symptoms were stable throughout the study (p > 0.06) with no serious adverse events related.DiscussionThis study highlights that plant-based nutrition support using a vegan-suitable plant-based ONS is highly complied with, improving the nutritional outcomes of patients at risk of malnutrition

Oljerelaterade miljögiftshalter i sediment och biota efter dieselspill i Himleån

Natten den 14 april 2022 skedde ett större dieselläckage efter en bränslestöld på fastigheten Varberg Signalen 14:1, Cylindervägen 12. Läckaget härrörde från en ovanjordscistern tillhörande Circle K. IVL Svenska Miljöinstitutet fick i uppdrag att undersöka drabbade vattenrecipienter, Himleån, Getteröns naturreservat och Farehammarsviken, med avseende på oljeföroreningsgrad och biologiskt viktiga komponenter. Uppdraget omfattade också att ge förslag på eventuella åtgärder och hur oljeutsläppet bör följas upp. Den 9–10 maj 2022 togs prover av sediment och biota på en rad platser längs Himleån, i Getteröns naturreservat och vid utloppet till Farehammarsviken. Dagvattenröret från området där olyckan skedde har sitt utlopp i ett mänskligt skapat biflöde till Himleån. I Himleån genomfördes provtagningen från utloppet ner till den punkt där ån rinner under väg 41 från strandkanten. På övriga platser längs ån provtogs sediment i åfåran med hjälp av Van Veen-huggare där detta var möjligt. Vid järnvägsbron (gränsen till naturreservatet) och vid Himleåns utlopp gjordes sedimentsprovtagningen från båt med Van Veen-huggare. Vid varje provtagningspunkt togs bilder och anteckningar för att dokumentera förhållandena på platsen. Vid fyra speciellt utvalda platser, varav en referenslokal uppströms olycksplatsen, genomfördes elfiske.Kemiska analyser av sedimentproverna visade på höga halter av alifater och PAH:er vid ett antal platser längs Himleån (provplats 2, 3 och 9) och i stora dammen inom Getteröns naturreservat (provplats 11, 14 och 16). I stora dammen var halterna höga främst i sediment från kanalen in mot den lilla dammen samt längs med stranden vid vägen. Utifrån de höga PAH-halter som påträffats i sedimenten vid dagvattenröret (provplats 2), men även med hänsyn till de förhöjda halter som påträffats vid andra provtagningsplatser i området, föreslogs uppföljande provtagning och analyser i dessa områden efter fågelsäsongen för att minimera störningarna. Resultaten från den uppföljande provtagningen skulle bidra till ett uppdaterat underlag för rekommendation om och var sanering bör utföras för att undvika eventuella biologiska effekter och vidare spridning av förorenat sediment. IVL rekommenderade också att absorptionslänsarna vid dagvattenutloppet skulle ligga kvar, åtminstone till resultaten från den uppföljande provtagningen analyserats för att kunna fånga upp lättare PAH:er. Analyser av fiskmuskel visade förhöjda halter PAH endast vid provplats 7 (Kockargården). Halterna överskred dock inte gränsvärdena för halter i fiskmuskel (HVMFS2019:25), men bör följas upp med ett senare elfiske för att utesluta följdeffekter av spill och åtgärder. Vid analyser av alger påvisades halter över referensprovet för PAH:er i samtliga prover. På tre platser (provplats 8, 10 och 16) överskreds gränsvärdet för kräftdjur och blötdjur (HVMFS 2019:25). Det saknas gränsvärden för vattenväxter. En uppföljande provtagning av gifter i sediment vid samma provplatser genomfördes av IVL 8 december 2022. Analyserna visade då för normaliserade värden inga kvarvarande detekterbara halter alifater eller PAH:er uppströms länsar vid dagvattenutloppet (provplats 2). Däremot hade halterna PAH nedströms länsar (provplats 3) ökat och översteg för floranten gränsvärde (HVMFS 2019:25). Även vid fågelskyddsområdet norr om dämmet (provplats 11) hade halterna PAH ökat och översteg för bens(g,h,i)perylen gränsvärde (HAV2018:31). IVL bedömer utifrån dessa resultat att provplats 3 efter länsar vid dagvattenutloppet bör saneras, gärna snarast innan våren tar fart. Här bör man ta kontakt med länsstyrelsen för att samråda kring lämpliga saneringsmetoder samt utarbeta en plan för efterkontroll. Analysresultat vid uppföljning av övriga provplatser föranleder inga åtgärder.Commissioned by Circle K Sverige AB.</p

Work from pub: The potential for co-working spaces in rural English pubs to improve business and community resilience

Between 2000 and 2017 the number of pubs in the UK declined by 26% and the COVID-19 pandemic exacerbated this, with six pubs demolished or converted each week. For rural areas the loss of pubs has been felt particularly strongly given the important role they play in rural hospitality and gastronomy, community cohesion and wellbeing. At the same time the pandemic has prompted a shift towards home and remote-working and the demand for co-working spaces has increased. Offering co-working spaces in rural pubs may help to reduce their vulnerability and strengthen community-orientated networks and the sense of identity within rural communities. This research uses face-to-face visits to pubs to better understand the barriers and enablers of providing co-working spaces to remote and home-workers. By talking to pub landlords, co-workers and the wider community is explores the validity of co-working as an opportunity to help rural pubs innovate and diversify their income and considers the implications for rural community resilience and meeting the needs of post-pandemic ways of working. The project focuses on the potential of pub co-working from a managerial, as well as a customer and community perspective to assess how pub ownership models (managed, tenanted, freehold and community owned) influence the motivation of managers to innovate and introduce new initiatives such as co-working space. The findings and lessons learned are distilled into a ‘Rural Co-working Pub Toolkit‘ which provides practical guidance to the pub industry, rural pub managers and those working in rural community development

Work from the Pub: The potential for pubs to offer workspaces for teleworkers in rural areas

Between 2000 and 2022 the number of pubs in the UK declined by 25% and the COVID-19 pandemic exacerbated this, with six pubs demolished or converted each week. For rural areas the loss of pubs has been felt particularly strongly given the important role they play in community cohesion and wellbeing. At the same time the pandemic has prompted a shift towards home working and the demand for remote workspaces has increased. Offering workspaces in rural pubs may help provide teleworkers with additional opportunities for co-working within their community and help reduce the sense of isolation associated with working from home as well as reducing the vulnerability of the pub. This research uses both qualitative and quantitative analysis to better understand the barriers and enablers of providing workspaces for teleworkers within pubs. By talking to pub landlords, trade associations and those working from home it explores the validity of offering a range of workspace options, including pub desks, work from pub packages, meeting spaces and conference facilities. Such provision is often limited in rural areas and offering these spaces may be an opportunity to help rural pubs innovate and diversify their income whilst also meeting the needs of post-pandemic ways of working. It is anticipated that findings and lessons will be distilled into a ‘Rural Co-working Pub Toolkit‘ to provide practical guidance to the pub industry, rural pub managers and those working in rural community development