bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myotube hypertrophy is associated with cancer-like metabolic reprogramming and limited by PHGDH
Authors: Stadhouders Lian E.M. , Verbrugge Sander A.J. , Smith Jonathon A.B. , Gabriel Brendan M. , Hammersen Tim D. , Kolijn Detmar , Vogel Ilse S.P. , Mohamed Abdalla D. , de Wit Gerard M.J. , Offringa Carla , Hoogaars Willem M. , Gehlert Sebastian , Wackerhage Henning , Jaspers Richard T. 
Published: December 2020

AbstractMuscle fiber size and oxidative metabolism are inversely related, suggesting that a glycolytic metabolism may offer a growth advantage in muscle fibers. However, the mechanisms underlying this advantage remains unknown. Nearly 100 years ago, Warburg reported that cancer cells take up more glucose to produce glycolytic intermediates for anabolic reactions such as amino acid-protein synthesis. The aim of this study was to test whether glycolysis contributes to anabolic signalling responses and hypertrophy in post-mitotic muscle cells. Skeletal muscle hypertrophy was induced in vitro by treating mouse C2C12 myotubes with IGF-1.14C glucose was added to differentiation medium and radioactivity in isolated protein was measured. We exposed differentiated C2C12 and primary mouse myotubes, to 2-deoxyglucose (2DG) and PHGDH siRNA upon which we assessed myotube diameter and signaling pathways involved in the regulation of muscle fiber size. Here, we present evidence that, hypertrophying C2C12 myotubes undergo a cancer-like metabolic reprogramming. First, IGF-1-induced C2C12 myotube hypertrophy increases shunting of carbon from glucose into protein. Second, reduction of glycolysis through 2-deoxy-D-glucose (2DG) lowers C2C12 and primary myotube size 16-40%. Third, reducing the cancer metabolism-associated enzyme PHGDH decreases C2C12 and primary myotube size 25-52%, whereas PHGDH overexpression increases C2C12 myotube size ≈20%. Fourth, the muscle hypertrophy-promoting kinase AKT regulates PHGDH expression. Together these results suggest that glycolysis is important for hypertrophying C2C12 myotubes by reprograming their metabolism similar to cancer cells.
Interactions Between SQUAMOSA and SVP MADS-box Proteins Regulate Meristem Transitions During Wheat Spike Development
Authors: Li Kun , Debernardi Juan M. , Li Chengxia , Lin Huiqiong , Zhang Chaozhong , Dubcovsky Jorge 
Published: December 2020

ABSTRACTA better understanding of spike development can contribute to improving wheat productivity. MADS-box genesVRN1andFUL2(SQUAMOSA-clade) play critical and redundant roles in wheat spike and spikelet development, where they act as repressors of MADS-box genes of theSHORT VEGETATIVE PHASE(SVP) clade (VRT2,SVP1andSVP3). Here, we show that wheatvrt2 svp1mutants are late flowering, have shorter stems, increased number of spikelets per spike and unusual axillary inflorescences in nodes of the elongating stem. Constitutive expression ofVRT2resulted in leafy glumes and lemmas, reversion of basal spikelets to spikes, and down-regulation of MADS-genes involved in floral development. Moreover, constitutive expression ofVRT2enhanced spikelet defects offul2, whereasvrt2reduced vegetative characteristics in the spikelets ofvrn1 ful2mutants heterozygous forVRN-A1. TheseSVP-SQUAMOSAgenetic interactions were paralleled by physical interactions among their encoded proteins. SVP proteins were able to reduce SQUAMOSA-SEPALLATA interactions in yeast-three-hybrid experiments. We propose that SQUAMOSA-SVP complexes act during the early reproductive phase to promote heading, formation of the terminal spikelet, and stem elongation, but that down-regulation ofSVPgenes is then necessary for the formation of SQUAMOSA-SEPALLATA complexes that are required for normal spikelet and floral development.
A small-scale shRNA screen in primary mouse macrophages identifies a role for the Rab GTPase Rab1b in controlling Salmonella Typhi growth
Authors: Solano-Collado Virtu , Colamarino Rosa Angela , Calderwood David A , Baldassarre Massimiliano , Spanò Stefania 
Published: December 2020

AbstractSalmonella Typhi, the causative agent of typhoid fever, a life-threatening systemic infection, is a human restricted bacterial pathogen. A fundamental aspect of S. Typhi pathogenesis is its ability to survive in human macrophages but not in macrophages from other animals (i.e. mice). Despite the importance of macrophages in establishing systemic S. Typhi infection, the mechanisms that macrophages use to control the growth of S. Typhi and the role of these mechanisms in the bacterium’s adaptation to the human host are mostly unknown. To facilitate unbiased identification of genes involved in controlling the growth of S. Typhi in macrophages, we report optimized experimental conditions required to perform loss-of function pooled shRNA screens in primary mouse bone-marrow derived macrophages. Following infection with a fluorescently-labeled S. Typhi, cells defective in genes important for controlling S. Typhi growth (and therefore unable to kill S. Typhi) are sorted based on the intensity of fluorescence (i.e. number of intracellular fluorescent bacteria). shRNAs enriched in the fluorescent population are identified by next-generation sequencing. A proof-of-concept screen targeting the mouse Rab GTPases confirmed Rab32 as important to restrict S. Typhi in mouse macrophages. Interestingly and rather unexpectedly, this screen also revealed that Rab1b controls S. Typhi growth in mouse macrophages. This constitutes the first report of a Rab GTPase other than Rab32 involved in S. Typhi host-restriction. The methodology described here should allow genome-wide screening to identify mechanisms controlling the growth of S. Typhi and other intracellular pathogens in primary immune cells.
GATA4/5/6 family transcription factors are conserved determinants of cardiac versus pharyngeal mesoderm fate
Authors: Song Mengyi , Yuan Xuefei , Racioppi Claudia , Leslie Meaghan , Aleksandrova Anastasiia , Christiaen Lionel , Wilson Michael D. , Scott Ian C. 
Published: December 2020

AbstractGATA4/5/6 transcription factors play essential, conserved roles in heart development. How these factors mediate the transition from multipotent mesoderm progenitors to a committed cardiac fate is unclear. To understand how GATA4/5/6 modulate cell fate decisions we labelled, isolated, and performed single-cell gene expression analysis on cells that expressgata5at pre-cardiac time points spanning gastrulation to somitogenesis. We found that most mesendoderm-derived lineages had dynamicgata5/6expression. In the absence of Gata5/6, the population structure of mesendoderm-derived cells was dramatically altered. In addition to the expected absence of cardiac mesoderm, we observed a concomitant expansion of cranial-pharyngeal mesoderm. Functional genetic analyses in zebrafish and the invertebrate chordateCiona, which possess a single GATA4/5/6 homolog, revealed an essential and cell-autonomous role for GATA4/5/6 in promoting cardiac and inhibiting pharyngeal mesoderm identity. Overall, the maintenance and repression of GATA4/5/6 activity plays a critical, evolutionarily conserved role in early development.
Identification of a novel cationic glycolipid inStreptococcus agalactiaethat contributes to brain entry and meningitis
Authors: Joyce Luke R. , Manzer Haider S. , Mendonça Jéssica da C. , Villarreal Ricardo , Nagao Prescilla E. , Doran Kelly S. , Palmer Kelli L. , Guan Ziqiang 
Published: December 2020

AbstractBacterial membrane lipids are critical for membrane bilayer formation, cell division, protein localization, stress responses, and pathogenesis. Despite their critical roles, membrane lipids have not been fully elucidated for many pathogens. Here, we report the discovery of a novel cationic glycolipid, Lysyl-Glucosyl-Diacylglycerol (Lys-Glc-DAG) that is synthesized in high abundance by the bacteriumStreptococcus agalactiae(Group BStreptococcus, GBS). To our knowledge, Lys-Glc-DAG is more positively charged than any other known lipids. Lys-Glc-DAG carries two positive net charges per molecule, distinct from the widely described lysylated phospholipid Lysyl-phosphatidylglycerol (Lys-PG) which carries one positive net charge due to the presence of a negatively charged phosphate moiety. We use normal phase liquid chromatography (NPLC) coupled with electrospray ionization (ESI) high-resolution tandem mass spectrometry (HRMS/MS) and genetic approaches to determine that Lys-Glc-DAG is synthesized by the enzyme MprF in GBS, which covalently modifies the neutral glycolipid Glc-DAG with the cationic amino acid lysine. GBS is a leading cause of neonatal meningitis, which requires traversal of the endothelial blood-brain barrier (BBB). We demonstrate that GBS strains lackingmprFexhibit a significant decrease in the ability to invade BBB endothelial cells. Further, mice challenged with a GBSΔmprFmutant developed bacteremia comparably to Wild-Type infected mice yet had less recovered bacteria from brain tissue and a lower incidence of meningitis. Thus, our data suggest that Lys-Glc-DAG may contribute to bacterial uptake into host cells and disease progression. Importantly, our discovery provides a platform for further study of cationic lipids at the host-pathogen interface.
Mobility shift-based electrophoresis coupled with fluorescent detection enables real-time enzyme analysis of carbohydrate sulfatase activity
Authors: Byrne Dominic P , London James A , Eyers Patrick A , Yates Edwin A , Cartmell Alan 
Published: December 2020

AbstractSulfated carbohydrate metabolism is a fundamental process, which occurs in all domains of life. Carbohydrate sulfatases are enzymes that remove sulfate groups from carbohydrates and are essential to the depolymerisation of complex polysaccharides. Despite their biological importance, carbohydrate sulfatases are poorly studied and challenges remain in accurately assessing the activity, specificity and kinetic parameters. Most notably, separation of desulfated products from sulfated substrates is currently a time-consuming process. In this paper, we describe the development of rapid capillary electrophoresis coupled to substrate fluorescence detection as a high-throughput and facile means of analysing carbohydrate sulfatase activity. The approach has utility for the determination of both kinetic and inhibition parameters and is based on existing microfluidic technology coupled to a new synthetic fluorescent 6S-GlcNAc carbohydrate substrate. Furthermore, we compare this technique in terms of both time and resources, to high performance anion exchange chromatography and NMR-based methods, which are the two current ‘gold standards’ for enzymatic carbohydrate sulfation analysis. Our study clearly demonstrates the advantages of mobility shift assays for the quantification of near real-time carbohydrate desulfation by purified sulfatases, and could support the search for small molecule inhibitors of these disease-associated enzymes.One sentence summarySulfatases remove sulfate groups from biomolecules; in this study we report a rapid and robust capillary electrophoresis assay for the quantification of carbohydrate desulfation.
Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
Authors: Makhija Suraj , Brown David , Bourke Struan , Wang Yina , Zhou Shuqin , Rudlaff Rachel , Cheloor-Kovilakam Rasmi , Huang Bo 
Published: December 2020

AbstractRecent advances in genome engineering have expanded our capabilities to study proteins in their natural states. In particular, the ease and scalability of knocking-in small peptide tags has enabled high throughput tagging and analysis of endogenous proteins. To improve enrichment capacities and expand the functionality of knock-ins using short tags, we developed the tag-assisted split enzyme complementation (TASEC) approach, which uses two orthogonal small peptide tags and their cognate binders to conditionally drive complementation of a split enzyme upon labeled protein expression. Using this approach, we have engineered and optimized the tag-assisted split HaloTag complementation system (TA-splitHalo) and demonstrated its versatile applications in improving the efficiency of knock-in cell enrichment, detection of protein-protein interaction, and isolation of biallelic gene edited cells through multiplexing.
Systematic research of e-learning platforms for solving challenges faced by Indian engineering students
Authors: Thakker Shivangi Viral , Parab Jayesh , Kaisare Shubhankar 
Published: December 2020

PurposeAs educational institutes began to address the challenges posed by COVID-19, e-learning came to the foreground as the best bet left. This study is in quest of revealing engineering student's perceptions of the available e-learning platforms, thus surfacing the underlying bottlenecks. Further, it aims at providing solutions that would help enhance the e-learning experience not only in pandemic times but also in the long run.Design/methodology/approachThis holistic research begins with a comprehensive comparative study about the available e-learning platforms, followed by a primary data analysis through an online survey of 364 engineering students from various colleges and branches. The collected data was analyzed to detect bottlenecks in online learning and suggestions are given for solving some challenges.FindingsOn a five-point Likert scale, the available e-learning platforms garnered ratings ranging from 2.81 to 3.46. Google meet was the most preferred platform. However, with a net promoter score (NPS) of 30.36, Microsoft Teams emerged as the most satisfying platform. Technical shortcomings clubbed with psychological and biological factors were found to be taking a toll on e-learning.Research limitations/implicationsThis innovative research is based on the perceptions of engineering students hailing majorly from Indian cities, and hence, it may be having educational stream bias and geographical bias. The research could be further extended to cover rural areas and global trends in e-learning.Originality/valueThe research offers a thorough analysis of e-learning platforms, as seen through the lens of engineering students. Furthermore, the analysis does not constrain itself to the technicalities and thus proves to be an all-encompassing one, potent enough to surface critical issues marring the e-learning experience.
Emerging living styles post-COVID-19: housing flexibility as a fundamental requirement for apartments in Jeddah
Authors: Bettaieb Donia M. , Alsabban Reem 
Published: December 2020

PurposeThe COVID-19 pandemic resulted in the compulsory quarantine of many of the world's inhabitants, and by staying at home, several functional developments emerged in residential spaces in Jeddah that affected the role of the house as a contributor to individuals' quality of life under the pressures of quarantine. Given the necessity of the apartments to adapt to these emerging developments, this study explores the determinants associated with the flexibility of residential apartments by looking at the extent to which they meet the new psychological, social and cultural roles required by their users post-COVID-19.Design/methodology/approachThe qualitative approach (1) extracted concepts related to the flexibility of housing from the available literature and (2) extrapolated the flexibility of the residential apartments from the participants' study (12 families) in different areas of Jeddah by analyzing the results of targeted interviews.FindingsThere is a gap in the participant's understanding of the quality of housing and the level of satisfaction with a housing design that differs before and after their quarantine experience. The participant's adaptation to self-quarantine was mainly through furniture distribution, and housing flexibility was less reliant on the physical transformation of the place than on the change in the inhabitants' perceptions. There was an indication that the deficiencies of flexibility in design relates to the functional, cultural and structural aspects of residential buildings.Originality/valueThis study generated suggestions to develop the foundations for flexible housing design and activate its role under the post-quarantine context according to social and cultural variables. Some proposals should become future requirements for residential apartments to benefit officials and stakeholders to develop housing flexibility.
Leadership in Brazilian public universities: initiatives conducted by three state universities of São Paulo in the context of COVID-19 pandemic
Authors: Anholon Rosley , Serafim Milena Pavan , Lourenzani Wagner Luiz , Silva Iris Bento , Rampasso Izabela Simon 
Published: December 2020

PurposeThe purpose of this paper is to present the role of aspects related to public leadership in the actions developed by three state universities in São Paulo (Brazil) in the context of the COVID-19 pandemic, namely the University of Campinas (Unicamp), University of São Paulo (USP) and São Paulo State University (Unesp).Design/methodology/approachSince this is a viewpoint article, part of the information presented is characterized by the authors' points of view. It should be highlighted, however, that the information provided is based on searches in scientific bases, institutional websites and published press reports.FindingsAccording to the authors, aspects of public leadership are being properly employed by the public servants of the analyzed universities, resulting in the positive actions that have been implemented.Originality/valueThere is no existing literature on public leadership in these Brazilian universities during the COVID-19 pandemic.
Shape sensing and feedback control of the catheter robot for interventional surgery
Authors: Qi Fei , Chen Bai , She Shigang , Gao Shuyuan 
Published: December 2020

Purpose
This paper aims to present a shape sensing method and feedback control strategy based on fiber Bragg grating (FBG) sensor to improve the control accuracy of the robot and ensure the safety of the cardiac interventional surgery.

Design/methodology/approach
To theoretically describe the shape of the catheter robot, the kinematic model is established by the geometric analysis method. And to obtain the actual shape, a large curvature assemble sensor based on FBG is adopted and a novel simple shape reconstruction model is proposed, which can provide the shape curve and distal position. In addition, the influence of external load on the bending deformation is investigated by experiments. To improve the shape accuracy of the robot, a shape feedback control method is presented to control the catheter robot, which can control the robot to bend into the pre-given desired shape.

Findings
Experiment results verify the effectiveness of the shape sensing method and the reconstruction model, and the correlation coefficients of three sets of curve in different coordinate directions are 0.9986, 0.9992 and 0.9999. Results of the shape feedback experiment show that the curvature error and direction angle error are 1.42% and 10.3%, respectively. The continuum catheter robot can be controlled to achieve the desired bending shape.

Originality/value
The shape reconstruction method and feedback control strategy proposed in this paper can improve the control accuracy of the robot to avoid the risk of the collision with the surrounding blood vessels, the tissues and organs.
Implementation challenges of COVID-19 safety measures at construction sites in South Africa
Authors: Amoah Christopher , Simpeh Fredrick 
Published: December 2020

Purpose
The COVID-19 pandemic drastically changed safety measures in every industry, including the construction industry. Thus, the construction companies have instituted safety measures at the construction sites to curve the disease’s spread among the workforce. This paper aims to examine the challenges encountered by construction firms in implementing COVID-19 safety measures at construction sites.

Design/methodology/approach
A qualitative research approach was adopted for this study using open-ended interview questions to solicit data from 19 construction professionals currently working on a construction project in South Africa. Content analysis with the assistance of an Excel spreadsheet was used to analyse the data collected.

Findings
The findings indicate that there are numerous challenges such as ignorance of COVID-19, the supply of poor personal protective equipment (PPEs) by contractors, lack of compliance, sanitising construction materials, difficulty in sharing tools and equipment, public transport usage by workers, superstition (COVID-19 is for a particular group of people), complying with social distancing rules, among others in the implementation of the COVID-19 safety measure at the construction site to curb the spread of the disease among the workers. These challenges have, therefore, hampered their effort to strictly adhere to the safety measures in accordance with the COVID-19 safety protocol at the project sites currently under construction.

Research limitations/implications
The interviewees were construction professionals working in the South African construction industry during the COVID-19 period.

Practical implications
The implication is that, due to the challenges faced in implementing the COVID-19 safety measures, workers on the construction site are not adequately protected from contracting COVID-19. The workers may thus contract the disease at the project sites and transmit it to their families and vice versa, which may have further implications on the spread of the disease within the communities and society.

Originality/value
The study has identified implementation challenges of the COVID-19 safety measures at construction sites of which the construction stakeholders must institute measures to overcome since COVID-19 has become part of our daily life. The study also recommends some preventive measures to the owners of construction companies to help overcome or minimise these COVID-19 safety implementation hurdles to minimise the spread of the disease among the construction site workers.
Creative tourism – providing the answers to a more inclusive society
Authors: Couret Caroline 
Published: December 2020

Purpose
This paper aims to provide an overview of the short-term impact of COVID19 on the new tourism paradigm worldwide, as well imagining how it could influence society’s inclusion. The purpose is thus to share some insight into this unprecedented situation.

Design/methodology/approach
This paper uses a practical and empirical approach, based on readings and conversations with international stakeholders, and contrasted with the Creative Tourism Network’s background.

Findings
This paper explores the impact of the COVID-19 crisis on the tourism industry, human values and intangibles and new opportunities for societal inclusion, together with examples of good practices.

Practical implications
The viewpoint is based on observation, analysis and conversations with stakeholders around the world. It also draws on examples and practical cases from the destinations’ members of the Creative Tourism Network.

Originality/value
This paper draws on observation and reflections about the evolution of societal inclusion through new forms of creative and niche tourism, accelerated by the COVID-19 crisis.