scholarly journals A functional genetic toolbox for human tissue-derived organoids

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Dawei Sun ◽  
Lewis Evans ◽  
Francesca Perrone ◽  
Vanesa Sokleva ◽  
Kyungtae Lim ◽  
...  
Keyword(s):  
Developmental Biology ◽  
Flow Cytometry ◽  
Disease Modelling ◽  
Endogenous Gene ◽  
Efficient Gene ◽  
Key Features ◽  
Foetal Lung ◽  
The Impact ◽  
Gene Perturbation ◽  
Crispr Activation

Human organoid systems recapitulate key features of organs offering platforms for modelling developmental biology and disease. Tissue-derived organoids have been widely used to study the impact of extrinsic niche factors on stem cells. However, they are rarely used to study endogenous gene function due to the lack of efficient gene manipulation tools. Previously, we established a human foetal lung organoid system (Nikolić et al., 2017). Here, using this organoid system as an example we have systematically developed and optimised a complete genetic toolbox for use in tissue-derived organoids. This includes 'Organoid Easytag' our efficient workflow for targeting all types of gene loci through CRISPR-mediated homologous recombination followed by flow cytometry for enriching correctly-targeted cells. Our toolbox also incorporates conditional gene knock-down or overexpression using tightly-inducible CRISPR interference and CRISPR activation which is the first efficient application of these techniques to tissue-derived organoids. These tools will facilitate gene perturbation studies in tissue-derived organoids facilitating human disease modelling and providing a functional counterpart to many on-going descriptive studies, such as the Human Cell Atlas Project.

Changing Work and Work-Related Travel and the Impact of Covid-19

2021 ◽  
Vol 47 (3) ◽  
pp. 336-354
Author(s):  
Kiron Chatterjee ◽  
Fiona Crawford
Keyword(s):  
Health Outcomes ◽  
Digital Age ◽  
Data Series ◽  
Work Related ◽  
Policy Interventions ◽  
Working Practices ◽  
Key Features ◽  
The Uk ◽  
Almost All ◽  
The Impact

The nature of work was undergoing dramatic change before the pandemic as the digital age continued to transform all sectors of society. In this paper we describe pre-pandemic trends in types of work, the workforce and working arrangements in the UK. We show how these changes were having gradual yet significant impacts on commuting and other work-related travel which were apparent in national travel data series. Key features of these impacts were increasing diversification and flexibility in work travel. We bring together findings on how working practices and travel have been altered by the pandemic and report expectations and opinions on its longer-term legacies. The pandemic has accelerated pre-pandemic trends and led to a shift in how work is performed for almost all sectors of the economy – but grasping the opportunity for this to contribute to deep carbon reductions from transport and to improve equity and health outcomes will require carefully directed policy interventions.

Candidiasis and the impact of flow cytometry on antifungal drug discovery

2017 ◽  
Vol 12 (11) ◽  
pp. 1127-1137
Author(s):  
Tsun Sheng N. Ku ◽  
Stella Bernardo ◽  
Carla J. Walraven ◽  
Samuel A. Lee
Keyword(s):  
Flow Cytometry ◽  
Drug Discovery ◽  
Antifungal Drug ◽  
The Impact

scholarly journals Talent Value Creation during Digital Transformation: A Inductive Model of HR Reengineering

2021 ◽  
Vol 4 (4) ◽  
Author(s):  
Hsiu-Hua Hu ◽  
Yaozong Zhu
Keyword(s):  
Qualitative Study ◽  
Value Creation ◽  
Talent Development ◽  
Digital Transformation ◽  
Four Dimensions ◽  
Research Questions ◽  
Key Features ◽  
The Impact ◽  
A Company ◽  
Insight Into

In this study, we are to explore (1) features of HR reengineering, (2) the impact of business digitalization strategies on digital transformation and HR engineering, (3) the impact of business digitalization strategies and HR reengineering on talent value creation, and present the results of a qualitative study that offers insight into 42 “thought units”, which were “categorizing” into four dimensions corresponding to our research questions: (1) plan, (2) do, (3) check, and (4) action. The “check” dimension corresponds to the four key features of HR reengineering related to business digitalization strategy, and how to create talent value when a company successfully implements business-led digital transformation, HR reengineering, and talent value creation, including (1) talent planning, (2) talent introduction, (3) talent adjustment, and (4) talent development.

scholarly journals Understanding axial progenitor biology in vivo and in vitro

Development ◽  
2021 ◽  
Vol 148 (4) ◽  
pp. dev180612
Author(s):  
Filip J. Wymeersch ◽  
Valerie Wilson ◽  
Anestis Tsakiridis
Keyword(s):  
Spinal Cord ◽  
Vertebral Column ◽  
Cell Types ◽  
Disease Modelling ◽  
Recent Developments ◽  
The Common ◽  
Key Features ◽  
Trunk Musculature

ABSTRACTThe generation of the components that make up the embryonic body axis, such as the spinal cord and vertebral column, takes place in an anterior-to-posterior (head-to-tail) direction. This process is driven by the coordinated production of various cell types from a pool of posteriorly-located axial progenitors. Here, we review the key features of this process and the biology of axial progenitors, including neuromesodermal progenitors, the common precursors of the spinal cord and trunk musculature. We discuss recent developments in the in vitro production of axial progenitors and their potential implications in disease modelling and regenerative medicine.

Australian Bills of Rights and the ‘New Commonwealth Model of Constitutionalism’

2013 ◽  
Author(s):  
Simon Evans ◽  
Julia Watson
Keyword(s):  
Human Rights ◽  
High Court ◽  
Human Rights Protection ◽  
Rights Protection ◽  
Human Rights Act ◽  
Human Rights Act 1998 ◽  
Key Features ◽  
The Uk ◽  
High Court Of Australia ◽  
The Impact

This chapter examines the influence of the new Commonwealth model of human rights protection (exemplified by the UK Human Rights Act 1998) on the form of the two Australian statutory Bills of Rights, and then considers the impact of Australia's distinctive legal culture and constitutional structure on the operation of these instruments. In particular, it examines the impact of culture and structure in the decision of the High Court of Australia in R. v Momcilovic [2011] HCA 34; (2011) 280 A.L.R. As a result of that case, key features of the Australian Bills of Rights now diverge from the dominant UK approach, a divergence so striking that it may no longer be possible to identify the Australian Bills of Rights as exemplars of the new Commonwealth model.

scholarly journals Zidovudine-Mediated Autophagy Inhibition Enhances Mitochondrial Toxicity in Muscle Cells

2018 ◽  
Vol 63 (1) ◽  
Author(s):  
H. Lin ◽  
M. V. Stankov ◽  
J. Hegermann ◽  
R. Budida ◽  
D. Panayotova-Dimitrova ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Therapeutic Drug ◽  
Ros Generation ◽  
Mtor Inhibition ◽  
Membrane Hyperpolarization ◽  
Thymidine Analogue ◽  
Drug Concentrations ◽  
Mitochondrial Functions ◽  
The Impact

ABSTRACT Nucleoside reverse transcriptase inhibitors (NRTI), such as zidovudine (AZT), are constituents of HIV-1 therapy and are used for the prevention of mother-to-child transmission. Prolonged thymidine analogue exposure has been associated with mitochondrial toxicities to heart, liver, and skeletal muscle. We hypothesized that the thymidine analogue AZT might interfere with autophagy in myocytes, a lysosomal degradation pathway implicated in the regulation of mitochondrial recycling, cell survival, and the pathogenesis of myodegenerative diseases. The impact of AZT and lamivudine (3TC) on C2C12 myocyte autophagy was studied using various methods based on LC3-green fluorescent protein overexpression or LC3 staining in combination with Western blotting, flow cytometry, and confocal and electron microscopy. Lysosomal and mitochondrial functions were studied using appropriate staining for lysosomal mass, acidity, cathepsin activity, as well as mitochondrial mass and membrane potential in combination with flow cytometry and confocal microscopy. AZT, but not 3TC, exerted a significant dose- and time-dependent inhibitory effect on late stages of autophagosome maturation, which was reversible upon mTOR inhibition. Inhibition of late autophagy at therapeutic drug concentrations led to dysfunctional mitochondrial accumulation with membrane hyperpolarization and increased reactive oxygen species (ROS) generation and, ultimately, compromised cell viability. These AZT effects could be readily replicated by pharmacological and genetic inhibition of myocyte autophagy and, most importantly, could be rescued by pharmacological stimulation of autophagolysosomal biogenesis. Our data suggest that the thymidine analogue AZT inhibits autophagy in myocytes, which in turn leads to the accumulation of dysfunctional mitochondria with increased ROS generation and compromised cell viability. This novel mechanism could contribute to our understanding of the long-term side effects of antiviral agents.

scholarly journals Histone deacetylase inhibitors valproic acid and vorinostat enhance trastuzumab-mediated antibody-dependent cell-mediated phagocytosis

2020 ◽  
Vol 8 (1) ◽  
pp. e000195 ◽  
Author(s):  
Johannes Laengle ◽  
Julijan Kabiljo ◽  
Leah Hunter ◽  
Jakob Homola ◽  
Sophie Prodinger ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Valproic Acid ◽  
Cancer Cells ◽  
Histone Deacetylase ◽  
Breast Cancer Cells ◽  
Histone Deacetylase Inhibitors ◽  
Combined Treatment ◽  
Dependent Cell ◽  
The Impact

BackgroundThe monoclonal antibody (mAb) trastuzumab is part of the standard of care for patients with human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer. Antibody-dependent cell-mediated phagocytosis (ADCP) and cytotoxicity (ADCC) are major mechanisms of action of the mAb trastuzumab. Histone deacetylase inhibitors (HDACi), such as valproic acid (VPA) or vorinostat (SAHA), exert several immunostimulatory properties, which contribute at least in part to their anticancer effect. However, the impact of HDACi-induced immunostimulatory effects on trastuzumab-mediated anti-tumor immune response is not well characterized.MethodsWe analyzed the ADCP and ADCC activity of peripheral blood mononuclear cells (PBMCs) from age and gender-matched healthy volunteers (n=5) against HDACi-treated HER2-overexpressing breast cancer cells (SKBR3), using a well-established in vitro three-color imaging flow cytometry and flow cytometry approach.ResultsVPA and SAHA enhanced trastuzumab-mediated ADCP and trastuzumab-independent cytotoxicity. Mechanistically, VPA upregulated the activating antibody-binding receptor Fc-gamma receptor (FcγR) IIA (CD32A) on monocytes (CD14+). Moreover, VPA and SAHA downregulated the anti-apoptotic protein myeloid leukemia cell differentiation 1 (MCL1) in breast cancer cells. Additionally, VPA and SAHA induced an immunogenic cell death, characterized by the exposure of calreticulin (CALR), as well as decreased the “do not eat me” signal CD47 on tumor cells.ConclusionsHDACi VPA and SAHA increase trastuzumab-mediated phagocytosis and trastuzumab-independent cytotoxicity. The immunomodulatory activities of those HDACi support a rationale combined treatment approach with mAb for cancer treatment.

scholarly journals Systemic anti-commensal response to fungi analyzed by flow cytometry is related to gut mycobiome ecology

Microbiome ◽  
2020 ◽  
Vol 8 (1) ◽  
Author(s):  
Alicia Moreno-Sabater ◽  
Gaelle Autaa ◽  
Delphine Sterlin ◽  
Amenie Jerbi ◽  
Remy Villette ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Alpha Diversity ◽  
Fungal Species ◽  
Individual Variability ◽  
Humoral Immune ◽  
Healthy Donors ◽  
Significant Difference ◽  
Genus Richness ◽  
Humoral Responses ◽  
The Impact

Abstract Background Interest for the study of gut mycobiota in relation with human health and immune homeostasis has increased in the last years. From this perspective, new tools to study the immune/fungal interface are warranted. Systemic humoral immune responses could reflect the dynamic relationships between gut mycobiota and immunity. Using a novel flow cytometry technology (Fungi-Flow) to determine immunoglobulin (Ig) responses to fungi, we studied the relationships between gut mycobiota and systemic humoral anti-commensal immunity. Results The Fungi-Flow method allows a sensitive and specific measurement of systemic IgG responses against 17 commensal and environmental fungi from the two main divisions; Ascomycota and Basidiomycota. IgG responses exhibited a high inter-individual variability. Anti-commensal IgG responses were contrasted with the relative abundance, alpha-diversity, and intra-genus richness of fungal species in gut mycobiota of twenty healthy donors. Categorization of gut mycobiota composition revealed two differentiated fungal ecosystems. Significant difference of anti-Saccharomyces systemic IgG responses were observed in healthy donors stratified according to the fungal ecosystem colonizing their gut. A positive and significant correlation was observed between the variety of IgG responses against fungal commensals and intestinal alpha-diversity. At the level of intra-genus species richness, intense IgG responses were associated with a low intra-genus richness for known pathobionts, but not commensals. Conclusions Fungi-Flow allows an easy and reliable measure of personalized humoral responses against commensal fungi. Combining sequencing technology with our novel Fungi-Flow immunological method, we propose that there are at least two defined ecosystems in the human gut mycobiome associated with systemic humoral responses. Fungi-Flow opens new opportunities to improve our knowledge about the impact of mycobiota in humoral anti-commensal immunity and homeostasis.

scholarly journals Pre-Strike Ballots and Collective Bargaining: The Impact of Quorum and Ballot Mode Requirements on Access to Lawful Industrial Action

2018 ◽  
Vol 48 (3) ◽  
pp. 343-376
Author(s):  
Breen Creighton ◽  
Catrina Denvir ◽  
Richard Johnstone ◽  
Shae McCrystal ◽  
Alice Orchiston
Keyword(s):  
Collective Bargaining ◽  
Empirical Research ◽  
Trade Union ◽  
Qualitative Empirical Research ◽  
Industrial Action ◽  
Two Factors ◽  
Research Findings ◽  
Key Features ◽  
The Uk ◽  
The Impact

Abstract In 2016, the UK Parliament passed the Trade Union Act 2016 (TU Act 2016), which introduced new quorum and approval requirements for pre-strike ballots. In Australia, mandatory pre-strike ballots, including a quorum requirement, were first introduced in 2006. This article explains the key features of the Australian pre-strike ballot system and reports on quantitative and qualitative empirical research findings on the operation of the ballots process to analyse the majority and quorum requirements, mode of ballot (postal, attendance or electronic) and choice of ballot agent. Quorum is the biggest obstacle to Australian unions authorising strike action under the pre-strike ballot rules, and postal ballots fail to reach quorum at significantly higher rates than do attendance ballots. By introducing quorums and retaining the requirement that all pre-strike ballots must be conducted by post, the TU Act 2016 endorsed the two factors under the Australian regime most likely to impede the authorisation of strike action in a pre-strike ballot.

Restructuring labour markets

2018 ◽  
pp. 33-54
Author(s):  
Anthony Lloyd
Keyword(s):  
Capital Accumulation ◽  
Historical Context ◽  
Labour Markets ◽  
Symbolic Order ◽  
Animal Spirits ◽  
On Demand ◽  
Key Features ◽  
Post War ◽  
Political Economic ◽  
The Impact

This chapter places the service economy in a political-economic and socio-historical context. The narrative details the emergence of a regulatory sleeve to limit the ‘animal spirits’ of capitalism’s non-ethical generative core. The post-war period of relative prosperity provides stability and a measurable sense of progress but ultimately gives way to neoliberalism at the end of the 1970s. The new ‘symbolic order’ shifts ground from security to flexibility in order to open up avenues for capital accumulation. Although successful in this respect, the impact upon labour markets and employment is significant and engenders a shift from industry and manufacturing to consumerism and service. The chapter outlines the impact of the financial crisis on labour markets and employment and demonstrates the growth of on-demand work. The key features of this chapter are the shift from security to flexibility and the composition of labour markets which now reflect imperatives of consumer culture but lack stability for employees.

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