bluuebunny/crossref_metadata_2025_split huggingface dataset:
Highly Efficient Knockin in Human iPS Cells and Rat Embryos by CRISPR/Cas9 Molecular Optimization
Authors: Chenouard Vanessa , Leray Isabelle , Tesson Laurent , Remy Séverine , Fortun Agnès , Bernardeau Karine , Cherifi Yacine , David Laurent , Anegon Ignacio 
Published: September 2021

AbstractThe CRISPR/Cas9 system is now the gold standard for the generation of genetically modified cell and animal models but knockin is a bottleneck. One reason could be that there is no consensus regarding the concentrations of its components to be used. Here, we defined optimal Cas9 protein, guide RNA and short donor DNA concentrations on a GFP to BFP conversion model of human induced pluripotent stem cells and point mutations on rat transgenic embryos. With a molecular rational approach of the CRISPR/Cas9 system and study of ribonucleoprotein complex formation by nanodifferential scanning fluorimetry, we defined that Cas9/guide RNA 1/1 molar ratio with 0.2μM and 0.4μM of Cas9, coupled with 2μM of ssODN are sufficient for optimal and high knockin frequencies in rat embryos and human induced pluripotent stem cells, respectively. These optimal conditions use lower concentrations of CRISPR reagents to form the RNP complex than most conditions published while achieving 50% of knockin. This study allowed us to reduce costs and toxicity while improving editing and knockin efficacy on two particularly key models to mimic human diseases.
WheatEARLY FLOWERING3is a dawn-expressed circadian oscillator component that regulates heading date
Authors: Wittern Lukas , Steed Gareth , Taylor Laura J. , Ramirez Dora Cano , Pingarron-Cardenas Gabriela , Gardner Keith , Greenland Andy , Hannah Matthew A. , Webb Alex A. R. 
Published: September 2021

AbstractUsing an eight-parent Multiparent Advanced Generation Inter-Cross (MAGIC) population we investigated how variation at circadian clock-associated genes contributes to the regulation of heading date in UK and European winter wheat varieties. We identified homoeologues ofEARLY FLOWERING3 (ELF3) as candidates for theEarliness per se(Eps)D1andB1 lociin field conditions. We confirmed that a SNP within the coding region ofTaELF3-B1is a candidate polymorphism underlying theEps-B1 locus.We found that a reported deletion at theEps-D1 locusencompassingTaELF3-D1,is instead a novel allele that lies within an introgression region containing an inversion relative to the Chinese Spring D genome. UsingT. turgidum cv. Kronoscarrying loss of function alleles ofTtELF3we show thatELF3does regulate heading by demonstrating that the loss of a singleELF3homoeologue was sufficient to alter heading date. These studies demonstrated thatELF3forms part of the circadian oscillator but loss of all homoeologues was required to affect circadian rhythms. Similarly, loss of functionalLUX ARRHYTHMO(LUX) inT. aestivum, an orthologue of a protein partner of Arabidopsis ELF3, severely disrupted circadian rhythms.ELF3andLUXtranscripts are not co-expressed at dusk suggesting the structure of the wheat circadian oscillator might differ to that of Arabidopsis. Our demonstration that alteration toELF3homoeologues can affect heading date separate from effects on the circadian oscillator suggests a role forELF3in cereal photoperiodic responses that could be selected for, without pleiotropic deleterious alterations to circadian rhythms.
Glucagon Receptor-mediated Regulation of Gluconeogenic Gene Transcription is Endocytosis-dependent in Primary Hepatocytes
Authors: Cajulao Jan Mikhale B. , von Zastrow Mark E. , Sanchez Erica L. 
Published: September 2021

AbstractA number of G protein-coupled receptors (GPCRs) are now thought to use endocytosis to promote cellular cAMP signaling that drives downstream transcription of cAMP-dependent genes. We tested if this is true for the Glucagon Receptor (GCGR), which mediates physiological regulation of hepatic glucose metabolism via cAMP signaling. We show that epitope-tagged GCGRs undergo clathrin and dynamin-dependent endocytosis in HEK293 cells after activation by glucagon, and transit via EEA1-marked endosomes shown previously to be sites of GPCR/Gs-stimulated production of cAMP. We further show that endocytosis potentiates cytoplasmic cAMP elevation produced by GCGR activation and promotes transcription of PCK1, the gene which encodes the enzyme catalyzing the rate-limiting step in gluconeogenesis. We verify endocytosis-dependent induction of PCK1 expression by endogenous GCGRs in primary hepatocytes, and show similar control of two other gluconeogenic genes (PGC1α and G6PC). Together, these results implicate the endosomal signaling paradigm in metabolic regulation by glucagon.
Recruitment of an ancient branching program to suppress carpel development in maize flowers
Authors: Klein Harry , Gallagher Joseph , Demesa-Arevalo Edgar , Abraham-Juárez María Jazmín , Heeney Michelle , Feil Regina , Lunn John E. , Xiao Yuguo , Chuck George , Whipple Clinton , Jackson David , Bartlett Madelaine 
Published: September 2021

AbstractFloral morphology is immensely diverse. One developmental process acting to shape this diversity is growth suppression. For example, grass flowers exhibit extreme diversity in floral sexuality, arising through differential suppression of stamens or carpels. In maize, carpels undergo programmed cell death in half of the flowers initiated in ears and in all flowers in tassels. The HD-ZIP I transcription factor geneGRASSY TILLERS1(GT1) is one of only a few genes known to regulate this process. To identify additional regulators of carpel suppression, we performed agt1enhancer screen, and found a genetic interaction betweengt1andramosa3(ra3). RA3is a classic inflorescence meristem determinacy gene that encodes a trehalose-6-phosphate (T6P) phosphatase (TPP). Dissection of floral development revealed thatra3single mutants have partially derepressed carpels, whereasgt1; ra3double mutants have completely derepressed carpels. Surprisingly,gt1suppressesra3inflorescence branching, revealing a role forgt1in meristem determinacy. Supporting these genetic interactions, GT1 and RA3 proteins colocalize to carpel nuclei in developing flowers. Global expression profiling revealed common genes misregulated in single and double mutant flowers, as well as in derepressedgt1axillary meristems. Indeed, we found thatra3enhancesgt1vegetative branching, similar to the roles for the trehalose pathway andGT1homologs in the eudicots. This functional conservation over ~160 million years of evolution reveals ancient roles forGT1-like genes and the trehalose pathway in regulating axillary meristem suppression, later recruited to mediate carpel suppression. Our findings expose hidden pleiotropy of classic maize genes, and show how an ancient developmental program was redeployed to sculpt floral form.
A novel widespread bacterial structure related to the flagellar type III secretion system
Authors: Kaplan Mohammed , Oikonomou Catherine M , Wood Cecily R. , Chreifi Georges , Ghosal Debnath , Dobro Megan J , Yao Qing , McDowall Alasdair , Briegel Ariane , Beeby Morgan , Chang Yi-Wei , Shaffer Carrie L. , Jensen Grant J. 
Published: September 2021

AbstractThe flagellar type III secretion system (fT3SS) is a suite of membrane-embedded and cytoplasmic proteins responsible for building the bacterial flagellar motility machinery. Homologous proteins form the injectisome machinery bacteria use to deliver effector proteins into eukaryotic cells, and other family members have recently been reported to be involved in the formation of membrane nanotubes. Here we describe a novel, ubiquitous and evolutionarily widespread hat-shaped structure embedded in the inner membrane of bacteria, of yet-unidentified function, that is related to the fT3SS, adding to the already rich repertoire of this family of nanomachines.
An Extended Motif in the SARS-CoV-2 Spike Modulates Binding and Release of Host Coatomer in Retrograde Trafficking
Authors: Dey Debajit , Singh Suruchi , Khan Saif , Martin Matthew , Schnicker Nicholas J. , Gakhar Lokesh , Pierce Brian G. , Hasan S. Saif 
Published: September 2021

Abstractβ-Coronaviruses such as SARS-CoV-2 hijack coatomer protein-I (COPI) for spike protein retrograde trafficking to the progeny assembly site in endoplasmic reticulum-Golgi intermediate compartment (ERGIC). However, limited residue-level details are available into how the spike interacts with COPI. Here we identify an extended COPI binding motif in the spike that encompasses the canonical K-x-H dibasic sequence. This motif demonstrates selectivity for αCOPI subunit. Guided by an in silico analysis of dibasic motifs in the human proteome, we employ mutagenesis and binding assays to show that the spike motif terminal residues are critical modulators of complex dissociation, which is essential for spike release in ERGIC. αCOPI residues critical for spike motif binding are elucidated by mutagenesis and crystallography and found to be conserved in the zoonotic reservoirs, bats, pangolins, camels, and in humans. Collectively, our investigation on the spike motif identifies key COPI binding determinants with implications for retrograde trafficking.
The statistical sensitivity of highway project cost to geo-heterogeneity in the tropics
Authors: Amadi Alolote 
Published: September 2021

PurposeThe study is carried out to analytically reconnoiter geotechnical index properties of subgrade soils as key variables that shape the cost profile of road infrastructure projects in a tropical geographic setting with starkly heterogenous ground conditions.Design/methodology/approachUsing the Niger Delta region, as a point of reference, data on geotechnical index properties of subgrade soils at spatially dispersed locations for 61 completed highway projects are collated. Exploratory statistical tests were carried out to infer significant associations with final project costs before regression analysis. Regression analysis is principally deployed as an explanatory analytical tool, relevant to quantify the sensitivity of highway project costs to the individual and collective impact of geotechnical variables.FindingsSeveral parameters of expansivity and compressibility exhibited significantly strong associations with the final costs recorded on the highway projects. The statistical analysis further established a cause-effect relationship, whereby small changes in the geotechnical properties of sub-grade soils at project locations, would result in disproportionately large changes in the cost of road construction.Practical implicationsThe study findings provide insight into the sensitivity of road construction costs to geotechnical variables, which can serve as a useful input in financial risk analysis for development appraisal and the generation of location adjustment factors.Originality/valueThe study statistically demonstrates location-induced construction cost profiles, triggered in response to the spatial geotechnical variability and occurrence of problem subgrade soils in the humid tropics, which may be different from those traditionally established in studies of cold and temperate climate soils.
Empowering vulnerable microfinance women through entrepreneurship: opportunities, challenges and the way forward
Authors: Ranabahu Nadeera , Tanima Farzana Aman 
Published: September 2021

PurposeThe purpose of this study is to explore how vulnerabilities arising from physical impairments, age, widowhood, forced displacement due to war or natural disasters and sexual orientation affect women’s microfinance-related entrepreneurial activities and economic empowerment.Design/methodology/approachThis paper is a structured literature review and uses preferred reporting items for systematic reviews and meta-analyses (PRISMA) method. The paper also uses agency, resources and achievement dimensions of empowerment to assess the findings.FindingsThis study reveals that although microfinance services promote women entrepreneurship, they also exacerbate exclusion and further discrimination. Individual-, household-, institutional- and community-level structures shape women’s agencies to access and use loans and conduct entrepreneurial activities; hence, ultimately economic empowerment.Originality/valueThe originality of this paper arises from the conceptual model we synthesised from our review which illustrates how vulnerable women’s agency is shaped by resources and how it affects their achievements in relation to entrepreneurial activities.
Industry 4.0 technologies usage: motives and enablers
Authors: Čater Tomaž , Čater Barbara , Černe Matej , Koman Matjaž , Redek Tjaša 
Published: September 2021

PurposeThe paper aims to contribute to a better understanding of the drivers for the use of Industry 4.0 technologies by investigating (1) what motivates companies to consider using I4 technologies and (2) what enables (or hinders) the intention to use I4 technologies to translate into their actual use.Design/methodology/approachThe study uses survey data collected from a sample of export-oriented manufacturing companies with more than 10 employees. Final analysis is conducted on 124 companies.FindingsThe results show that companies are proactively approaching I4. Only efficiency motives and expected competitive advantage have a positive effect on the intention to use I4 technologies, which in turn positively influences their actual use. The external, legitimacy-based, motives do not play a significant role in explaining the intention to use. With respect to I4 technology enablers, employee competency positively moderates and availability of finance negatively moderates the relationship between intention to use and actual use.Research limitations/implicationsThe work extends the existing knowledge base on I4 technology drivers in companies that are not major global trendsetters but are heavily embedded in the value chains of companies from the most industrially developed economies. The study is limited to manufacturing companies in a small European economy and should be retested in other contexts.Practical implicationsThe study can help managers implement I4 technologies in their companies more successfully.Originality/valueWe take a novel research approach by proposing a framework that clearly distinguishes between motives and enablers for the use of I4 technologies.
Manufacturing internationalization: from distance to proximity? A longitudinal analysis of offshoring choices
Authors: Podrecca Matteo , Orzes Guido , Sartor Marco , Nassimbeni Guido 
Published: September 2021

PurposeThis paper aims to offer a long-term systematic picture of the evolution of manufacturing offshoring (in terms of intensity, geography and drivers) highlighting the changes in the surrounding context and the resulting transitions points (“points in time”) that have shaped its development path.Design/methodology/approachThree statistical tools were adopted on a dataset of 644 cases. First, the authors resorted to multiple structural change tests to identify the transition points. Second, the authors explored offshoring geography by conducting a network analysis. Finally, the authors adopted gravity models to shed light on offshoring drivers.FindingsResults highlight three offshoring phases: expansion (2002–2006), reconsideration (2007–2009) and rationalization (2010 onwards). During the first phase, characterized by economic growth, firms were mainly interested in economic savings; offshoring to low-cost countries was the prevailing location strategy. Subsequently, during the economic crisis, the number of cases declined and the main drivers became market-based factors together with the research for cost savings. Finally, in the third phase, when the economy was still stagnating and new manufacturing technologies appeared, the number of offshoring cases has further decreased, and technological- and market-based factors have become the main location drivers.Originality/valueThe study is the first to adopt a systematic, empirical and quantitative approach to analyze the evolution of the manufacturing offshoring considering both the phenomenon itself and the triggering changes in the surrounding context. In doing this, the authors also tested the importance of considering the point in time in offshoring strategies.
Do repetitive and non-repetitive companies equally benefit from Lean 4.0?
Authors: Cifone Fabiana Dafne , Portioli Staudacher Alberto 
Published: September 2021

PurposeThe integration between the traditional lean management and Industry 4.0, namely called Lean 4.0, is under the spotlight of both academia and practitioners. While we agree on the benefits Lean 4.0 may bring to companies performance, we still lack a deep understanding of the characteristics of this paradigm, such as its effective application space. Recalling traditional lean better suits repetitive companies, we are keen to understand whether the anew Lean 4.0 will enlarge its application space.Design/methodology/approachWe performed an exploratory study, using a quantitative analysis based on Bayesian network approach to investigate whether Lean 4.0 results to be as effective in repetitive companies as in non-repetitive ones, in terms of operational performance.FindingsWhile our findings confirm that Lean 4.0 will enhance companies' performance regardless their production strategies, companies adopting a repetitive strategy do benefit from a much higher improvement. Our findings provide an insight on the true applicability space of Lean 4.0, which seems to be the same as the traditional lean.Originality/valueWe contribute to the ongoing debate of Lean 4.0 providing initial empirical evidence on how to improve the operational performance in non-repetitive companies, seeing that Lean 4.0 might not be the best choice in its current format.
Fans behave as buyers? Assimilate fan-based and team-based drivers of fan engagement
Authors: Fathy Doaa , Elsharnouby Mohamed H. , AbouAish Ehab 
Published: September 2021

PurposeCustomer engagement, as one form of interactive marketing, enhances organisational performance, in terms of sales growth, superior competitive advantage and increased profitability, particularly within the sports context. This research aims to explore fans' engagement behaviours with their sports teams and identify its drivers and outcomes.Design/methodology/approachThe researchers deployed mixed methods in this study via three phases: (1) A judgmental sampling technique, along with snowballing, were used to conduct in-depth interviews with twenty-two football fans, for the exploratory phase; (2) A convenience sample was also used for the quantitative phase, which was divided into two stages, (1) the pretesting stage (30 fans), and (2) the main data collection stage (407 fans) and (3) A judgmental sampling technique was applied for the qualitative validation phase (10 interviews with experts and practitioners).FindingsQualitative and quantitative results supported team jealousy, team competitiveness and team morality as new predictors for fan engagement behaviours. Further, while the fan role readiness had the most positive effect on management cooperation, team identification had the most predicting power for prosocial behaviour. Finally, team morality had the most significant positive impact on performance tolerance.Originality/valueDespite the considerable practical attention, and the recent extensive research, paid towards conceptualising customer engagement behaviours in the last decade, there is still a need for further exploration on the fan engagement concept to better understand fans' unique behavioural responses; accordingly, the current research was conducted.
Recent advances in the relationship between economic development and carbon emissions
Authors: Yin Ziqi , Jin Xue 
Published: September 2021

PurposeWith the rapid development of the economy, carbon emissions have also risen sharply. This study explores the relationship between the two by combining the literature of relevant fields and maps the analytical framework from the knowledge base to the research frontier model using CiteSpace.Design/methodology/approachUsing CiteSpace and data statistical tools, we conducted a bibliometric and visual analysis of nearly ten thousand research papers on carbon emissions and economic development published in the Web of Science (WOS) and China National Knowledge Infrastructure (CNKI) databases from 1991 to 2021.FindingsIt shows that research on economic development and carbon emissions is developing steadily and involves a wide range of fields. Notably, keywords such as “carbon emissions,” “economic growth,” and “energy consumption” had high frequency, centrality, and persistence. “carbon emissions,” “economic growth,” and “energy consumption” had high frequency, centrality, and persistence. Research institutions in the USA and China have made great contributions to research on economic development and carbon emissions. The authors should continue to enrich and improve research on related subjects and concerns to reasonably plan the path of carbon emission reduction and economic development.Originality/valueThe study analyzes the evolution of the relationship between carbon emissions and economic growth to provide scholars a more comprehensive and in-depth understanding of the relationship from an international perspective.
Alternative food sources interfere with removal of a fungal amphibian pathogen by zooplankton
Authors: Deknock Arne , Pasmans Frank , van Leeuwenberg Robby , Van Praet Sarah , Bruneel Stijn , Lens Luc , Croubels Siska , Martel An , Goethals Peter 
Published: September 2021

Abstract

While the amphibian disease chytridiomycosis is causing ongoing population declines and biodiversity losses around the globe, efficient mitigation strategies are lacking. The free‐living zoospores of the causative agents of this disease, the chytrid pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal), are a potential food source for filter‐feeding micropredators as part of the aquatic food web. While consumption of zoospores can lower environmental pathogen loads, alternative food sources may interfere with pathogen removal rates.

We compared the ability of three filter‐feeding zooplankton species, the cladoceran Daphnia magna, the rotifer Brachionus calyciflorus and the ostracod Heterocypris incongruens, to remove Bd zoospores in water and investigated the effect of alternative food sources, the green algae Pseudokirchneriella subcapitata and Chlorella vulgaris, on zoospore ingestion by D. magna.

Daphnia magna was the only micropredator candidate that effectively removed Bd zoospores from its environment, with an average removal rate of 1,012 ± 542 GE ind.−1 hr−1 within our test system. High concentrations (1 × 105 cells/ml) of large and easily ingestible P. subcapitata reduced pathogen removal rates, whereas the small and less edible C. vulgaris did not interfere with pathogen removal.

Synthesis and applications. We showed that Daphnia spp., which are keystone species in all sorts of aquatic habitats worldwide, are promising target agents for biologically mitigating chytridiomycosis infections and how natural food sources may interfere with this strategy. We also suggest potential management actions for biological disease mitigation, aiming to optimize environmental conditions for these target filter feeders, thereby reducing pathogen densities and eventually infection pressure in amphibian hosts. Examples of such management actions include, but are not limited to, removal of planktivorous fish, habitat restoration, nutrient control or agrochemical regulation in the vicinity of amphibian breeding ponds. Further studies, including field trials, are needed to confirm the effects of pathogen consumption on infection dynamics in natural situations and investigate the impact of intervention actions.