bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structure, immunogenicity, and conformation-dependent receptor binding of the post-fusion human metapneumovirus F protein
Authors: Huang Jiachen , Chopra Pradeep , Liu Lin , Nagy Tamas , Murray Jackelyn , Tripp Ralph A. , Boons Geert-Jan , Mousa Jarrod J. 
Published: May 2021

AbstractHuman metapneumovirus (hMPV) is an important cause of acute viral respiratory infection. As the only target of neutralizing antibodies, the hMPV fusion (F) protein has been a major focus for vaccine development and targeting by drugs and monoclonal antibodies (mAbs). While X-ray structures of trimeric pre-fusion and post-fusion hMPV F proteins from genotype A, and monomeric pre-fusion hMPV F protein from genotype B have been determined, structural data for the post-fusion conformation for genotype B is lacking. We determined the crystal structure of this protein and compared the structural differences of post-fusion hMPV F between hMPV A and B genotypes. We also assessed the receptor binding properties of the hMPV F protein to heparan sulfate. A library of heparan sulfate (HS) oligomers was used to verify the HS binding activity of hMPV F, and several compounds showed binding to predominantly pre-fusion hMPV F, but had limited binding to post-fusion hMPV F. Furthermore, mAbs to antigenic sites III and the 66-87 intratrimeric epitope block heparan binding. In addition, we evaluated the efficacy of post-fusion hMPV B2 F protein as a vaccine candidate in BALB/c mice. Mice immunized with hMPV B2 post-fusion F protein showed a balanced Th1/Th2 immune response and generated neutralizing antibodies against both subgroup A2 and B2 hMPV strains, which protected the mice from hMPV challenge. Antibody competition analysis revealed the antibodies generated by immunization target two known antigenic sites (III and IV) on hMPV F. Overall, this study provides new characteristics of the hMPV F protein informative for vaccine and therapy development.ImportanceHuman metapneumovirus (hMPV) is an important cause of viral respiratory disease. In this paper, we report the X-ray crystal structure of the hMPV fusion (F) protein in the post-fusion conformation from genotype B. We also assessed binding of the hMPV F protein to heparin and heparan sulfate, a previously reported receptor for the hMPV F protein. Furthermore, we determined the immunogenicity and protective efficacy of post-fusion hMPV B2 F protein, which is the first study using a homogenous conformation of the protein. Antibodies generated in response to vaccination give a balanced TH1/TH2 response and target two previously discovered neutralizing epitopes.
Protein detection in blood with single-molecule imaging
Authors: Mao Chih-Ping , Wang Shih-Chin , Su Yu-Pin , Tseng Ssu-Hsueh , He Liangmei , Wu Annie A. , Roden Richard B.S. , Xiao Jie , Hung Chien-Fu 
Published: May 2021

AbstractThe ability to identify and characterize individual biomarker protein molecules in patient blood samples could enable diagnosis of diseases at an earlier stage, when treatment is typically more effective. Single-molecule imaging offers a promising approach to accomplish this goal. However, thus far single-molecule imaging methods have only been used to monitor protein molecules in solutions or cell lysates, and have not been translated into the clinical arena. Furthermore, the detection limit of these methods has been confined to the picomolar (10−12 M) range. In many diseases, the circulating concentrations of biomarker proteins fall several orders of magnitude below this range. Here we describe Single-Molecule Augmented Capture (SMAC), a single-molecule imaging technique to visualize, quantify, and characterize individual protein molecules of interest down to the subfemtomolar (<10−15 M) range, even in complex biologic fluids. We demonstrate SMAC in a wide variety of applications with human blood samples, including the analysis of disease-associated secreted proteins, membrane proteins, and rare intracellular proteins. Using ovarian cancer as a model, a lethal malignancy in which high-grade disease is driven almost universally by alterations in the TP53 gene and frequently only diagnosed at a late, incurable stage, we found that mutant pattern p53 proteins are released into the blood in patients at an early stage in disease progression. SMAC opens the door to the application of single-molecule imaging in non-invasive disease profiling and allows for the analysis of circulating mutant proteins as a new class of highly specific disease biomarkers. The SMAC platform can be adapted to multiplex or high-throughput formats to characterize heterogeneous biochemical and structural features of circulating proteins-of-interest.One Sentence SummaryA single-molecule imaging approach detects individual disease-specific protein molecules, including mutant intracellular proteins, in blood samples.
Circularly permuted LOV domain as an engineering module for optogenetic tools
Authors: Geng Lequn , Shen Jiaqi , Wang Wenjing 
Published: May 2021

AbstractWe re-engineered a commonly-used light-sensing protein, LOV domain, using a circular permutation strategy to allow photoswitchable control of the C-terminus of a peptide. We demonstrate that the use of circularly permuted LOV domain on its own or together with the original LOV could expand the engineering capabilities of optogenetic tools.
Near-infrared imaging in fission yeast by genetically encoded biosynthesis of phycocyanobilin
Authors: Sakai Keiichiro , Kondo Yohei , Fujioka Hiroyoshi , Kamiya Mako , Aoki Kazuhiro , Goto Yuhei 
Published: May 2021

ABSTRACTNear-infrared fluorescent protein (iRFP) is a bright and stable fluorescent protein with excitation and emission maxima at 690 nm and 713 nm, respectively. Unlike the other conventional fluorescent proteins such as GFP, iRFP requires biliverdin (BV) as a chromophore because iRFP originates from bacteriophytochrome. Here, we report that phycocyanobilin (PCB) functions as a brighter chromophore for iRFP than BV, and biosynthesis of PCB allows live-cell imaging with iRFP in the fission yeastSchizosaccharomyces pombe. We initially found that fission yeast cells did not produce BV, and therefore did not show any iRFP fluorescence. The brightness of iRFP attached to PCB was higher than that of iRFP attached to BVin vitroand in fission yeast. We introduced SynPCB, a previously reported PCB biosynthesis system, into fission yeast, resulting in the brightest iRFP fluorescence. To make iRFP readily available in fission yeast, we developed an endogenous gene tagging system with iRFP and all-in-one integration plasmids, which contain genes required for the SynPCB system and the iRFP-fused marker proteins. These tools not only enable the easy use of iRFP in fission yeast and the multiplexed live-cell imaging in fission yeast with a broader color palette, but also open the door to new opportunities for near-infrared fluorescence imaging in a wider range of living organisms.
Plasmid-encoded H-NS controls extracellular matrix composition in a modern Acinetobacter baumannii urinary isolate
Authors: Benomar Saida , Di Venanzio Gisela , Feldman Mario F. 
Published: May 2021

ABSTRACTAcinetobacter baumannii is emerging as a multidrug-resistant (MDR) nosocomial pathogen of increasing threat to human health worldwide. The recent MDR urinary isolate UPAB1 carries the plasmid pAB5, a member of a family of large conjugative plasmids (LCP). LCP encode several antibiotic resistance genes and repress the type VI secretion system (T6SS) to enable their dissemination, employing two TetR transcriptional regulators. Furthermore, pAB5 controls the expression of additional chromosomally encoded genes, impacting UPAB1 virulence. Here we show that a pAB5-encoded H-NS transcriptional regulator represses the synthesis of the exopolysaccharide PNAG and the expression of a previously uncharacterized three-gene cluster that encodes a protein belonging to the CsgG/HfaB family. Members of this protein family are involved in amyloid or polysaccharide formation in other species. Deletion of the CsgG homolog abrogated PNAG production and Cup pili formation, resulting in a subsequent reduction in biofilm formation. Although this gene cluster is widely distributed in Gram-negative bacteria, it remains largely uninvestigated. Our results illustrate the complex cross-talks that take place between plasmids and the chromosomes of their bacterial host, which in this case can contribute to the pathogenesis of Acinetobacter.IMPORTANCEThe opportunistic human pathogen Acinetobacter baumannii displays the highest reported rates of multidrug resistance among Gram-negative pathogens. Many A. baumannii strains carry large conjugative plasmids like pAB5. In recent years, we have witnessed an increase in knowledge about the regulatory cross-talks between plasmids and bacterial chromosomes. Here we show that pAB5 controls the composition of the bacterial extracellular matrix, resulting in a drastic reduction in biofilm formation. The association between biofilm formation, virulence, and antibiotic resistance is well-documented. Therefore, understanding the factors involved in the regulation of biofilm formation in Acinetobacter has remarkable therapeutic potential.
Quantitative Tyr Phosphoproteomic Analyses Identify Cholesterol as a Master Regulator of the Tumor Adaptive Resistance to MAPK Inhibition
Authors: Wang Xu-Dong , Kim Chiho , Zhang Yajie , Rindhe Smita , Cobb Melanie H. , Yu Yonghao 
Published: May 2021

AbstractAlthough targeted inhibition of the MAPK pathway has achieved remarkable patient responses in many cancers with MAPK hyperactivation, the development of resistance has remained a critical challenge. Besides genomic resistance mechanisms, adaptive tumor response also underlies the resistance to targeted MAPK inhibitors. It is being increasingly appreciated that such bypass mechanisms often lead to the activation of many pro-survival kinases, which complicates the rational design of combination therapies. Here we performed global tyrosine phosphoproteomic (pTyr) analyses and demonstrated that targeted inhibition of MAPK signaling in melanoma cells leads to a profound remodeling of the pTyr proteome. Intriguingly, many of these kinases contain a cholesterol binding motif, suggesting that altered cholesterol metabolism might drive, in a coordinated fashion, the activation of these kinases. Indeed, we found a dramatic accumulation of intracellular cholesterol in melanoma cells (with BRAFV600E mutations) and non-small cell lung cancer cells (with KRASG12C mutations) treated with MAPK and KRASG12C inhibitors, respectively. Importantly, depletion of cholesterol not only prevented the MAPK inhibition-induced feedback activation of pTyr singling but also enhanced the cytotoxic effects of MAPK inhibitors, both in vitro and in vivo. Taken together, our findings provide the evidence suggesting that cholesterol functions as a master regulator of the tumor adaptive response to targeted MAPK inhibitors. These results also suggest that MAPK inhibitors could be combined with cholesterol-lowering agents to achieve a more complete and durable response in tumors with hyperactive MAPK signaling.
Systematic Tuning of Rhodamine Spirocyclization for Super-Resolution Microscopy
Authors: Lardon Nicolas , Wang Lu , Tschanz Aline , Hoess Philipp , Tran Mai , D’Este Elisa , Ries Jonas , Johnsson Kai 
Published: May 2021

AbstractRhodamines are the most important class of fluorophores for applications in live-cell fluorescence microscopy. This is mainly because rhodamines exist in a dynamic equilibrium between a fluorescent zwitterion and a non-fluorescent but cell-permeable spirocyclic form. Different imaging applications require different positions of this dynamic equilibrium, which poses a challenge for the design of suitable probes. We describe here how the conversion of the ortho-carboxy moiety of a given rhodamine into substituted acyl benzenesulfonamides and alkylamides permits the systematic tuning of the equilibrium of spirocyclization with unprecedented accuracy and over a large range. This allows to transform the same rhodamine into either a highly fluorogenic and cell-permeable probe for live-cell stimulated emission depletion (STED) microscopy, or into a spontaneously blinking dye for single molecule localization microscopy (SMLM). We used this approach to generate differently colored probes optimized for different labeling systems and imaging applications.
V-ATPase is a universal regulator of LC3 associated phagocytosis and non-canonical autophagy
Authors: Hooper Kirsty M. , Jacquin Elise , Li Taoyingnan , Goodwin Jonathan M. , Brumell John H. , Durgan Joanne , Florey Oliver 
Published: May 2021

AbstractNon-canonical autophagy is a key cellular pathway in immunity, cancer and neurodegeneration, characterised byConjugation ofATG8 to endolysosomalSingle-Membranes (CASM). CASM is activated by engulfment (endocytosis, phagocytosis), agonists (STING, TRPML1) and infection (influenza), dependent on the ATG16L1 WD40-domain, and specifically K490. However, the factor(s) associated with non-canonical ATG16L1 recruitment, and CASM induction, remain unknown. Here, we investigate a role for V-ATPase during non-canonical autophagy. We report that increased V0-V1 engagement is associated with, and sufficient for, CASM activation. Upon V0-V1 binding, V-ATPase directly recruits ATG16L1, via K490, during LC3-associated phagocytosis (LAP), STING- and drug-induced CASM, indicating a common mechanism. Furthermore, during LAP, key molecular players, including NADPH oxidase/ROS, converge on V-ATPase. Finally, we show that LAP is sensitive toSalmonellaSopF, which disrupts the V-ATPase-ATG16L1 axis, and provide evidence that CASM contributes to theSalmonellahost response. Together, these data identify V-ATPase as a universal regulator of CASM, and indicate that SopF evolved in part to evade non-canonical autophagy.
Translation-dependent mRNA localization toCaenorhabditiselegans adherens junctions
Authors: Tocchini Cristina , Rohner Michèle , Von Stetina Stephen E. , Mango Susan E. 
Published: May 2021

AbstractmRNA localization is an evolutionarily widespread phenomenon that facilitates sub-cellular protein targeting. Extensive work has focused on mRNA targeting through “zip codes” within untranslated regions (UTRs), while much less is known about translation-dependent cues. Here, we examine mRNA localization inCaenorhabditis elegansembryonic epithelia. From an smFISH-based survey, we identified mRNAs associated with the cell membrane or cortex, and with apical junctions in a stage- and cell type-specific manner. Mutational analyses for one of these transcripts,dlg-1/discs large, revealed that it relied on a translation-dependent process and did not require its 5’ or 3’ UTR. We suggest a model in whichdlg-1transcripts are co-translationally colocalized with the encoded protein: first the translating complex goes to the cell membrane through sequences of the SH3 domain, and then to the apical junction by the L27 and PDZ sequences. In addition, the Hook and GuK sequences contribute to the second step: they are required for mRNA, but not protein, to accumulate at the apical junctions from locations at or near the membrane. These studies identify a translation-based process for mRNA localization within developing epithelia and determine the necessary cis-acting sequences fordlg-1mRNA targeting.Summary statementAn smFISH-based survey identified a subset of mRNAs coding for junctional components that localize at or in the proximity of the adherent junction through a translation-dependent mechanism.
From satisfaction to loyalty: the role of emotional structures in the process of transition from satisfaction to loyalty
Authors: Ghorbanzadeh Davood 
Published: May 2021

PurposeSatisfaction and loyalty as vital and strategic concepts in marketing literature are highly important to companies and marketers. The review of the existing literature reveals a gap of the role of emotional constructs that can begin in a regular and rational sequence of satisfaction and ultimately lead to the formation of consumer loyalty. Hence, in the current study, we seek to answer the question of whether emotional constructs such as emotional attachment and love play a mediating role in the process of transitioning from satisfaction to loyalty in the correct sequence.Design/methodology/approachA total of 300 valid questionnaires on Smartphone and apparel brands were collected from respondents and analyzed using the partial least squares method.FindingsThe results showed that brand love is the strongest antecedent of brand loyalty and is the only variable that directly influences brand loyalty in comparison to satisfaction and emotional brand attachment. Moreover, the examination of indirect effects revealed that our assumption based on that the emotional structures such as emotional attachment and brand love play a mediating role in the process of transitioning from satisfaction to loyalty in a correct sequence is supported.Research limitations/implicationsGeneralizability needs to be established with a wider range of consumer groups. The survey was conducted in Iran and future research should assess the same product categories in other cultural settings as well as consider other product categories to assess the external validity of these results. The insights on consumers' brand relationships help brand managers devise effective brand management strategies.Practical implicationsThe managerial implications can guide managers toward enhancing the consumers' loyalty to the brand through a better understanding of the consumer loyalty process to a brand as well as better relational marketing practices.Originality/valueThe study validates the mediating role of emotional brand attachment and brand love in the relationship between brand satisfaction and brand loyalty; is one of the first to develop a conceptual model that examines the role of emotional structures in the process of transition from satisfaction to loyalty.; is one of few studies to develop the role of emotional structures in the form of a relational chain of brands in the process of transition from satisfaction to loyalty.
DMart an ace in India’s retail space
Authors: Mittal Ankur , Jain Anshul , Dhingra Tarun 
Published: May 2021

Learning outcomes
The instructor can use this for covering the concept of competitive advantage and valuation. To explore the relationship between superior profitability with business model and identify the source of competitive advantage. Determining the value of business through discounted cash flow (DCF) approach.

Case overview/synopsis
This case is based on the growth path of a retail chain company, DMart (Avenue Supermarket Private Limited), in the rapidly growing organized retail industry in India. It operates a unique business model that garners significant revenue growth with high profitability. The case covers the competitive advantages of DMart with respect to its peers and its valuation through the DCF model.

Complexity academic level
The case is suitable for teaching basic courses in corporate valuation and strategy at the post-graduate level. The following courses can also make use of this case: financial management, corporate finance and strategic management in emerging markets.

Supplementary materials
Teaching Notes are available for educators only.

Subject code
CSS 1: Accounting and Finance.
The Turkish Super League meets its first female club president
Authors: Ünsal Efe , Kaptanoğlu Sanem , Kabasakal Hayat 
Published: May 2021

Learning outcomes
The learning outcomes are as follows. First, female managers can face gender inequity in different forms, such as the glass ceiling and glass cliff, while they run for leadership positions, especially in male-dominated business contexts. Second, managers can show high performance and be effective leaders as long as they are aware of that all managers are evaluated according to a wide variety of criteria, and play many different managerial roles, such as interpersonal, informational and decisional roles. Finally, managers should pay attention to all stakeholders’ demands in decision-making process for sustainability and performing better in the long run.

Case overview/synopsis
Since football began gaining popularity in Turkey at the dawn of the 20th century, the sport remains the most popular national sport today. However, recently, a new name has shaken the world of Turkish football: Berna Gozbasi, the first female football manager in Turkish history. In the middle of 2019–2020 football season, Gozbasi became the first female club president after she assumed leadership of Kayserispor. Kayserispor was officially founded as a Turkish professional football club in 1966, and, as its name suggests, was based in Kayseri, a sizeable industrialised city located in Central Anatolia. The team competes in the “Turkish Super League”, Turkey’s top football competition. In this case, to discuss gender inequity, leadership, and management in the sport context, the authors explained the dilemma Gozbasi faced while she decided whether or not to accept this challenging role. Then, the authors examined the experiences she gained as a leader and the dilemma she faced to diminish the negative impact of the COVID-19 pandemic on the organisation she led.

Complexity academic level
Undergraduate and MBA students.

Supplementary materials
Teaching Notes are available for educators only. Please contact your library to gain login details or email support@emeraldinsight.com to request teaching notes.

Subject code
CSS 6: Human Resource Management.
The role of social media in promoting citizenship values of international students during the COVID-19 global health crisis
Authors: Oweis Thelal Iqab 
Published: May 2021

Purpose
The purpose of this study, accordingly, is to not only offer insights about Arab international students’ stress management and adjustment mechanisms while abroad but also bring to light the extent to which these digital citizens’ use of social media networks reinforces their citizenship values and preserves their national identity during the COVID-19 global health crisis.

Design/methodology/approach
The researcher adopted the descriptive approach, which is the appropriate scientific method for this type of research studies, grounded on an examination of the reality or phenomenon, depending on its quantitative analysis and its accurate description by means of collecting data, using the measurement tool of questionnaire to identify the role of social media in relation to citizenship during the COVID-19 global health crisis.

Findings
Jordanian exchange students heavily rely on social media during the pandemic, choosing Facebook as their most favorable choice during the pandemic to connect with family and friends near and far, while choosing WhatsApp as the most favorable choice for promoting citizenship values during the pandemic, with an average willingness to actually discuss citizenship through social media. The findings affirm results and conclusions of previous studies about the crucial role of social media in international students’ cross-cultural experiences, especially during stressful events like the COVID-19 global health crisis.

Originality/value
Unfortunately, and even before the outbreak of the COVID-19 global health crisis, Arab international students, including exchange students, are already underrepresented in studies of international students’ adjustment and stress management in hosting countries (Farrugia et al., 2017; Meleis, 1982). This marginalization, or partial exclusion, of Arab international students’ experiences abroad should alert scholars of missed research opportunities that should alter existing knowledge about international students, especially when we realize, for example, that Arab international students showed the highest stress level among international students – including Asians who, according to previous studies, usually struggle the most (Bai, 2016:103).