bluuebunny/crossref_metadata_2025_split huggingface dataset:
Skeletal dysplasia-causing TRPV4 mutations suppress the hypertrophic differentiation of human iPSC-derived chondrocytes
Authors: Dicks Amanda R. , Maksaev Grigory I. , Harissa Zainab , Savadipour Alireza , Tang Ruhang , Steward Nancy , Liedtke Wolfgang , Nichols Colin G. , Wu Chia-Lung , Guilak Farshid 
Published: June 2021

ABSTRACTMutations in the TRPV4 ion channel can lead to a range of skeletal dysplasias. However, the mechanisms by which TRPV4 mutations lead to distinct disease severity remain unknown. Here, we use CRISPR-Cas9-edited human induced pluripotent stem cells (hiPSCs) harboring either the mild V620I or lethal T89I mutations to elucidate the differential effects on channel function and chondrogenic differentiation. We found that hiPSC-derived chondrocytes with the V620I mutation exhibited increased basal currents through TRPV4. However, both mutations showed more rapid calcium signaling with a reduced overall magnitude in response to TRPV4 agonist GSK1016790A compared to wildtype. There were no differences in overall cartilaginous matrix production, but the V620I mutation resulted in reduced mechanical properties of cartilage matrix later in chondrogenesis. mRNA sequencing revealed that both mutations upregulated several anterior HOX genes and downregulated antioxidant genes CAT and GSTA1 throughout chondrogenesis. BMP4 treatment upregulated several essential hypertrophic genes in WT chondrocytes; however, this hypertrophic maturation response was inhibited in mutant chondrocytes. These results indicate that the TRPV4 mutations alter BMP signaling in chondrocytes and prevent proper chondrocyte hypertrophy, as a potential mechanism for dysfunctional skeletal development. Our findings provide potential therapeutic targets for developing treatments for TRPV4-mediated skeletal dysplasias.
Evolving small-molecule biosensors with improved performance and reprogrammed ligand specificity using OrthoRep
Authors: Javanpour Alex A. , Liu Chang C. 
Published: June 2021

AbstractGenetically-encoded biosensors are valuable for the optimization of small-molecule biosynthesis pathways, because they transduce the production of small-molecule ligands into a readout compatible with high-throughput screening or selection in vivo. However, engineering biosensors with appropriate response functions and ligand specificities remains challenging. Here, we show that the continuous hypermutation system, OrthoRep, can be effectively applied to evolve biosensors with high dynamic range, reprogrammed activity towards desired non-cognate ligands, and proper operational range for coupling to biosynthetic pathways. In particular, we encoded the allosteric transcriptional factor, BenM, on OrthoRep such that propagation of host yeast cells resulted in BenM’s rapid and continuous diversification. When these cells were subjected to cycles of culturing and sorting on BenM activity in the presence and absence of its cognate ligand, muconic acid, or the non-cognate ligand, adipic acid, we obtained multiple BenM variants that respond to their corresponding ligands. These biosensors outperform previously-engineered BenM-based biosensors by achieving substantially greater dynamic range (up to ~180-fold-induction) and broadened operational range. Expression of select BenM variants in the presence of a muconic acid biosynthetic pathway demonstrated sensitive biosensor activation without saturating response, which should enable pathway and host engineering for higher production of muconic and adipic acids. Given the streamlined manner in which high-performance and versatile biosensors were evolved using OrthoRep, this study provides a template for generating custom biosensors for metabolic pathway engineering and other biotechnology goals.
Macrophages stimulate epicardial VEGFaa expression to trigger cardiomyocyte proliferation in larval zebrafish heart regeneration
Authors: Bruton Finnius A. , Kaveh Aryan , Ross-Stewart Katherine M. , Matrone Gianfranco , Oremek Magdalena E.M. , Solomonidis Emmanouil G. , Tucker Carl S. , Mullins John J. , Brittan Mairi , Taylor Jonathan M. , Rossi Adriano G. , Denvir Martin A. 
Published: June 2021

AbstractCardiac injury induces a sustained macrophage response in both zebrafish and mammals. Macrophages perform a range of both beneficial and detrimental functions during mammalian cardiac repair, yet their precise roles in zebrafish cardiac regeneration are not fully understood. Here we characterise cardiac regeneration in the rapidly regenerating larval zebrafish laser injury model and use macrophage ablation and macrophage-null irf8 mutants to define the role of macrophages in key stages of regeneration. Live heartbeat-synchronised imaging and RNA sequencing revealed an early proinflammatory phase, marked by tnfa+ macrophages, which then resolved to an anti-inflammatory, profibrotic phase. Macrophages were required for cardiomyocyte proliferation but not for functional or structural recovery following injury. Importantly, we found that macrophages are specifically recruited to the epicardial-myocardial niche, triggering the expansion of the epicardium which upregulates VEGFaa expression to induce cardiomyocyte proliferation. Hence, revealing a novel mechanism by which macrophages facilitate cardiac regeneration.
Transcriptional reprogramming and constitutive PD-L1 expression in melanoma are associated with dedifferentiation and activation of interferon and tumor necrosis factor signaling pathways
Authors: Ahn Antonio , Rodger Euan J. , Gimenez Gregory , Stockwell Peter A. , Parry Matthew , Hersey Peter , Chatterjee Aniruddha , Eccles Michael R. 
Published: June 2021

AbstractMelanoma is the most aggressive type of skin cancer, with increasing incidence worldwide. Advances in targeted therapy and immunotherapy have improved the survival of melanoma patients experiencing recurrent disease, but unfortunately treatment resistance frequently reduces patient survival. Resistance to targeted therapy is associated with transcriptomic changes, and has also been shown to be accompanied by increased expression of programmed death ligand 1 (PD-L1), a potent inhibitor of immune response. Intrinsic upregulation of PD-L1 is associated with genome-wide DNA hypomethylation and widespread alterations in gene expression in melanoma cell lines. However, an in-depth analysis of the transcriptomic landscape of melanoma cells with intrinsically upregulated PD-L1 expression is lacking. To determine the transcriptomic landscape of intrinsically upregulated PD-L1 expression in melanoma, we investigated transcriptomes in melanomas with constitutive versus inducible PD-L1 expression (referred to as PD-L1CON and PD-L1IND). RNA-Seq analysis was performed on seven PD-L1CON melanoma cell lines and ten melanoma cell lines with low inducible PD-L1IND expression. We observed that PD-L1CON melanoma cells had a reprogrammed transcriptome with a characteristic pattern of dedifferentiated gene expression, together with active interferon (IFN) and tumor necrosis factor (TNF) signalling pathways. Furthermore, we identified key transcription factors that were also differentially expressed in PD-L1CON versus PD-L1IND melanoma cell lines. Overall, our studies describe transcriptomic reprogramming of melanomas with PD-L1CON expression.Simple SummaryMelanoma, an aggressive form of skin cancer, is frequently associated with drug resistance in the advanced stages. For instance, frequently resistance is observed to sequential treatment of melanoma with targeted therapy and immunotherapy. In this research, the authors investigated whether potential transcriptional mechanisms and pathways associated with PD-L1 protein expression could underlie targeted therapy drug resistance in melanoma. The authors found a PD-L1 expression transcriptional pattern underlies resistance to targeted therapy in a subgroup of melanomas. These melanomas were markedly dedifferentiated, as compared to melanomas that were not drug resistant. Understanding changes in transcription and molecular pathways that lead to drug resistance could allow researchers to develop interventions to prevent drug resistance from occurring in melanoma, which could also be relevant to other cancer types.
In vivo augmentation of a complex gut bacterial community
Authors: Cheng Alice G. , Ho Po-Yi , Jain Sunit , Meng Xiandong , Wang Min , Yu Feiqiao Brian , Iakiviak Mikhail , Brumbaugh Ariel R. , Nagashima Kazuki , Zhao Aishan , Patil Advait , Atabakhsh Katayoon , Weakley Allison , Yan Jia , Higginbottom Steven , Neff Norma , Sonnenburg Justin L. , Huang Kerwyn Casey , Fischbach Michael A. 
Published: June 2021

ABSTRACTEfforts to model the human gut microbiome in mice have led to important insights into the mechanisms of host-microbe interactions. However, the model communities studied to date have been defined or complex but not both, limiting their utility. In accompanying work, we constructed a complex synthetic community (104 strains, hCom1) containing the most common taxa in the human gut microbiome. Here, we used an iterative experimental process to improve hCom1 by filling open metabolic and/or anatomical niches. When we colonized germ-free mice with hCom1 and then challenged it with a human fecal sample, the consortium exhibited surprising stability; 89% of the cells and 58% of the taxa derive from the original community, and the pre- and post-challenge communities share a similar overall structure. We used these data to construct a second version of the community, adding 22 strains that engrafted following fecal challenge and omitting 7 that dropped out (119 strains, hCom2). In gnotobiotic mice, hCom2 exhibited increased stability to fecal challenge and robust colonization resistance against pathogenic Escherichia coli. Mice colonized by hCom2 versus human feces are similar in terms of microbiota-derived metabolites, immune cell profile, and bacterial density in the gut, suggesting that this consortium is a prototype of a model system for the human gut microbiome.
The respiratory supercomplex from C. glutamicum
Authors: Moe Agnes , Kovalova Terezia , Król Sylwia , Yanofsky David J. , Bott Michael , Sjöstrand Dan , Rubinstein John L. , Högbom Martin , Brzezinski Peter 
Published: June 2021

AbstractCorynebacterium glutamicum is a preferentially aerobic Gram-positive bacterium belonging to the Actinobacteria phylum, which also includes the pathogen Mycobacterium tuberculosis. In the respiratory chain of these bacteria, complexes III (CIII) and IV (CIV) form a CIII2CIV2 supercomplex that catalyzes oxidation of menaquinol and reduction of dioxygen to water. Electron transfer within the CIII2CIV2 supercomplex is linked to transmembrane proton translocation, which maintains an electrochemical proton gradient that drives ATP synthesis and transport processes. We isolated the C. glutamicum supercomplex and used cryo-EM to determine its structure at 2.9 Å resolution. The structure shows a central CIII2 dimer flanked by a CIV on each side. One menaquinone is bound in each of the QN and QP sites in each CIII, near the cytoplasmic and periplasmic sides, respectively. In addition, we identified a menaquinone positioned ~14 Å from heme bL on the periplasmic side. A di-heme cyt. cc subunit provides an electronic connection between each CIII monomer and the adjacent CIV. In CIII2, the Rieske iron-sulfur (FeS) proteins are positioned with the iron near heme bL. Multiple subunits interact to form a convoluted sub-structure at the cytoplasmic side of the supercomplex, which defines a novel path that conducts protons into CIV.
Human embryoid bodies as a novel system for genomic studies of functionally diverse cell types
Authors: Rhodes Katherine , Barr Kenneth A , Popp Joshua M , Strober Benjamin J , Battle Alexis , Gilad Yoav 
Published: June 2021

AbstractMost disease-associated loci, though located in putatively regulatory regions, have not yet been confirmed to affect gene expression. One reason for this could be that we have not examined gene expression in the most relevant cell types or conditions. Indeed, even large-scale efforts to study gene expression broadly across tissues are limited by the necessity of obtaining human samples post-mortem, and almost exclusively from adults. Thus, there is an acute need to expand gene regulatory studies in humans to the most relevant cell types, tissues, and states. We propose that embryoid bodies (EBs), which are organoids that contain a multitude of cell types in dynamic states, can provide an answer. Single cell RNA-sequencing now provides a way to interrogate developmental trajectories in EBs and enhance the potential to uncover dynamic regulatory processes that would be missed in studies of static adult tissue. Here, we examined the properties of the EB model for the purpose mapping inter-individual regulatory differences in a large variety of cell types.
The Catholic Church abuse scandal in Ireland: two steps forward, one step back by Pope Francis?
Authors: Pang Augustine , Hogan Eada , Andrasevic Igor 
Published: June 2021

PurposeIreland is viewed as the shining base for Catholicism. That image is shattered as survivors revealed the abuse in the Magdalene Laundries and Mother and Baby Homes, and sexual abuse by priests. This study aims to examine image repair efforts by the Pope during his August 2018 visit.Design/methodology/approachExamined against the Letter of His Holiness released days earlier, this study evaluates all the Pope's speeches during his visit to Ireland using the image repair theory (Benoit and Pang, 2008) as its theoretical lens.FindingsPope Francis used the evasion of responsibility strategy to address the Magdalene Laundries and Mother and Baby Homes scandal and denial, corrective action and mortification for sex abuse crisis.Research limitations/implicationsAddresses call by Ferguson et al. (2018) to examine the consistency and effectiveness of strategies.Practical implicationsBeyond rhetoric, stakeholders would be looking to organizational leaders to provide relief and concrete steps to recover from their pain.Originality/valueA leader's narratives represent the organization's narratives; thus, insights from this study can help leaders plan what they should say when conducting image repair. It is not just their own reputations that are on the line but, in this case, it is also the reputations of the people they represent
Bridging the gender digital divide: an analysis of existing guidance for gender digital inclusion programmes’ evaluations
Authors: Pawluczuk Alicja , Lee JeongHyun , Gamundani Attlee Munyaradzi 
Published: June 2021

Purpose
This aim of this paper is to examine the existing gender digital inclusion evaluation guidance and proposes future research recommendations for their evaluation. Despite modern progress in towards gender equality and women’s empowerment movements, women’s access to, use of and benefits from digital technologies remain limited owing to economic, social and cultural obstacles. Addressing the existing gender digital divide is critical in the global efforts towards the United Nations’ Sustainable Development Goals (SDGs). In recent years, there has been a global increase of gender digital inclusion programmes for girls and women; these programmes serve as a mechanism to learn about gender-specific digital needs and inform future digital inclusion efforts. Evaluation reports of gender digital inclusion programmes can produce critical insights into girls’ and women’s learning needs and aspirations, including what works and what does not when engaging girls and women in information and communications technologies. While there are many accounts highlighting the importance of why gender digital inclusion programmes are important, there is limited knowledge on how to evaluate their impact.

Design/methodology/approach
The thematic analysis suggests three points to consider for the gender digital inclusion programmes evaluation: context-specific understanding of gender digital inclusion programmes; transparency and accountability of the evaluation process and its results; and tensions between evaluation targets and empowerment of evaluation participants.

Findings
The thematic analysis suggests three points of future focus for this evaluation process: context-specific understanding of gender digital inclusion programmes; transparency and accountability of the evaluation process and its results; and tensions between evaluation targets and empowerment of evaluation participants.

Originality/value
The authors propose recommendations for gender digital inclusion evaluation practice and areas for future research.
Well-being and career change intention: COVID-19’s impact on unemployed and furloughed hospitality workers
Authors: Chen Chun-Chu (Bamboo) , Chen Ming-Hsiang 
Published: June 2021

Purpose
This study aims to examine the psychological distress experienced by unemployed and furloughed hospitality workers during the COVID-19 crisis and further investigate how this distress affects their career change intentions.

Design/methodology/approach
Derived from a sample of 607 unemployed and furloughed hospitality workers during the COVID-19 pandemic, the data for this research are analyzed using structural equation modeling.

Findings
This study reveals that unemployed and furloughed hospitality workers are financially strained, depressed, socially isolated and panic-stricken due to the pandemic’s effects. These effects lead to impaired well-being and an increased intention to leave the hospitality industry. Female and younger employees are impacted to a greater extent, while furloughed workers received fewer impacts compared to their laid-off compatriots.

Research limitations/implications
This study suggests that lockdown restrictions need to be implemented more deliberately, and the psychological well-being of the hospitality workforce deserves more immediate and continuing attention. It advises that hospitality businesses consider furloughs over layoffs when workforce reduction measures are necessary to combat the financial crisis.

Originality/value
This study adds to the current literature by examining the impact of the COVID-19 pandemic from the employee perspective. New insights are offered on the psychological toll of workforce reduction strategies during the financial fallout and how these distressing experiences affect career change intention.
Forging a collective entrepreneurial identity within existing organizations through corporate venturing
Authors: Zhang Ying , Biniari Marina G. 
Published: June 2021

PurposeThis study unpacks how organizational members construct a collective entrepreneurial identity within an organization and attempt to instill entrepreneurial features in the organization's existing identity.Design/methodology/approachThe study draws on the cases of two venturing units, perceived as entrepreneurial groups within their respective parent companies. Semi-structured interviews and secondary data were collected and analyzed inductively and abductively.FindingsThe data revealed that organizational members co-constructed a “corporate entrepreneur” role identity to form a collective shared belief and communities of practice around what it meant to act as an entrepreneurial group within their local corporate context and how it differentiated them from others. Members also clustered around the emergent collective entrepreneurial identity through sensegiving efforts to instill entrepreneurial features in the organization's identity, despite the tensions this caused.Originality/valuePrevious studies in corporate entrepreneurship have theorized on the top-down dynamics instilling entrepreneurial features in an organization's identity, but have neglected the role of bottom-up dynamics. This study reveals two bottom-up dynamics that involve organizational members' agentic role in co-constructing and clustering around a collective entrepreneurial identity. This study contributes to the middle-management literature, uncovering champions' identity work in constructing a “corporate entrepreneur” role identity, with implications for followers' engagement in constructing a collective entrepreneurial identity. This study also contributes to the organizational identity literature, showing how tensions around the entrepreneurial group's distinctiveness may hinder the process of instilling entrepreneurial features in an organization's identity.
Coaching to develop teacher professional noticing: planning with students and mathematics in mind
Authors: Jakopovic Paula M. 
Published: June 2021

PurposeThis paper examines how intentional mathematics coaching practices can develop teacher professional noticing of “ambitious teaching practices” (NCTM, 2020) through connected, collaborative coaching cycles.Design/methodology/approachNarrative analysis is used to examine observations of a mathematics coach and novice teacher to better understand the role of the coach in helping teachers attend to ambitious mathematics teaching (AMT) practices.FindingsThe initial findings of this study suggest that intentional use of focused goals, iterative coaching cycles and a gradual release model of coaching can support shifts in noticing of AMT from being led by the coach to being facilitated by the teacher.Originality/valueThis study offers new insights into the functions of mathematics coaching that can foster shifts in teacher noticing and practice toward AMT. It contributes to the literature on what mathematics coaching looks and sounds like in the context of conversations with teachers, as well as the potential influence that structured, intentional, ongoing coaching supports can have on teacher noticing.