bluuebunny/crossref_metadata_2025_split huggingface dataset:
Discovery of DTX3L inhibitors through a homogeneous FRET-based assay that monitors formation and removal of poly-ubiquitin chains
Authors: Vela-Rodríguez Carlos , Scarpulla Ilaria , Ashok Yashwanth , Lehtiö Lari 
Published: March 2023

AbstractUbiquitination is a complex and reversible protein post-translational modification in which the subsequent action of enzymes belonging to three different families, broadly referred to as E1, E2 and E3, results in the covalent linking of ubiquitin to a target protein. While this linkage is canonically an isopeptide bond between the C-terminus of ubiquitin and the lysine residue of the target protein, Ser, Thr, and Tyr can also be susceptible to ubiquitination through an oxyester bond. Once ubiquitinated, multiple units of ubiquitin can be attached to the initial ubiquitin thus extending it to a chain of ubiquitins. Ubiquitination regulates multiple cellular processes, but it is best known as a modification that targets proteins for proteasomal degradation following the formation poly-ubiquitin chains linked through lysine 48 or 63 of ubiquitin. Dysregulation of ubiquitination has been associated with multiple types of cancer and efforts have been carried out to develop technologies that lead to the identification of inhibitors of the enzymes involved in the ubiquitination cascade. Herein, we present the development of a FRET-based assay that allows us to monitor auto-ubiquitination of DTX3L, a RING-type E3 ubiquitin ligase. Our method shows a robust signal window with a robust average Z’ factor of 0.76. From a validatory screening experiment we have identified the first molecules that inhibit DTX3L with potencies in the low micromolar range. Additionally, we have expanded the system to study deubiquitinases such as USP28 that lead to reduction of FRET due to hydrolysis of fluorescent poly-Ub chains.
Structural basis of Cas12a R-loop propagation on pathway to DNA cleavage
Authors: Strohkendl Isabel , Moy Catherine , Nguyen Alexander-Hoi , Russell Rick , Taylor David W. 
Published: March 2023

AbstractCas12a is a CRISPR RNA-guided nuclease that engages target DNA through protein-DNA and RNA-DNA contacts. Initial PAM binding by Cas12a leads to formation of a 20bp R-loop between the complementary crRNA guide and target strand. Following specificity-determining R-loop formation, both DNA strands undergo RuvC-mediated cleavage. Current structures of Cas12a bound to its target only show the R-loop after formation, leaving an important gap in knowledge as to how Cas12a accommodates the extending R-loop and coordinates R-loop formation with nuclease activation. Here, we use cryo-EM to capture a series of kinetically trapped Cas12a R-loop intermediates and observe how Cas12a delivers each DNA strand into the RuvC active site. We show that Cas12a first interrogates target DNA via a 5bp seed, followed by dramatic Rec domain conformational flexibility to accommodate R-loop extension. Only during formation of the final R-loop base pairs do the Rec and BH domains engage in the majority of contacts with the R-loop. R-loop completion leads the nontarget strand to displace the RuvC lid and kink into the active site via a base stacking interaction. Following nontarget strand cleavage, we observe substantial Rec2 and Nuc domain dynamics as the TS is brought to the RuvC active site. Our kinetics-guided structural snapshots provide a comprehensive model describing Cas12a DNA targeting and highlight mechanistic differences between Cas12a and Cas9.
High-throughput, fluorescent-aptamer-based measurements of steady-state transcription rates forMycobacterium tuberculosisRNA polymerase
Authors: Jensen Drake , Manzano Ana Ruiz , Rector Maxwell , Tomko Eric J. , Record M. Thomas , Galburt Eric A. 
Published: March 2023

AbstractThe first step in gene expression is the transcription of DNA sequences into RNA. Regulation at the level of transcription leads to changes in steady-state concentrations of RNA transcripts, affecting the flux of downstream functions and ultimately cellular phenotypes. Changes in transcript levels are routinely followed in cellular contexts via genome-wide sequencing techniques. However,in vitromechanistic studies of transcription have lagged with respect to throughput. Here, we describe the use of a real-time, fluorescent-aptamer-based method to quantitate steady-state transcription rates of theMycobacterium tuberculosisRNA polymerase. We present clear controls to show that the assay specifically reports on promoter-dependent, full-length RNA transcription rates that are in good agreement with the kinetics determined by gel-resolved, α-32P NTP incorporation experiments. We illustrate how the time-dependent changes in fluorescence can be used to measure regulatory effects of nucleotide concentrations and identity, RNAP and DNA concentrations, transcription factors, and antibiotics. Our data showcase the ability to easily perform hundreds of parallel steady-state measurements across varying conditions with high precision and reproducibility to facilitate the study of the molecular mechanisms of bacterial transcription.Significance StatementRNA polymerase transcription mechanisms have largely been determined fromin vitrokinetic and structural biology methods. In contrast to the limited throughput of these approaches,in vivoRNA sequencing provides genome-wide measurements but lacks the ability to dissect direct biochemical from indirect genetic mechanisms. Here, we present a method that bridges this gap, permitting high-throughput fluorescence-based measurements ofin vitrosteady-state transcription kinetics. We illustrate how an RNA-aptamer-based detection system can be used to generate quantitative information on direct mechanisms of transcriptional regulation and discuss the far-reaching implications for future applications.
TimeTeller: a tool to probe the circadian clock as a multigene dynamical system
Authors: Vlachou Denise , Veretennikova Maria , Usselmann Laura , Vasilyev Vadim , Ott Sascha , Bjarnason Georg A. , Dallmann Robert , Levi Francis , Rand David A. 
Published: March 2023

AbstractRecent studies have established that the circadian clock influences onset, progression and therapeutic outcomes in a number of diseases including cancer and heart diseases. Therefore, there is a need for tools to measure the functional state of the molecular circadian clock and its downstream targets in patients. Moreover, the clock is a multi-dimensional stochastic oscillator and there are few tools for analysing it as a system. In this paper we consider the methodology behind Time-Teller, a machine learning tool that analyses the clock as a system and aims to estimate circadian clock function from a single transcriptome by modelling the multi-dimensional state of the clock. We demonstrate its potential for clock systems assessment by applying it to mouse, baboon and human microarray and RNA-seq data and show how to visualise and quantify the global structure of the clock, quantitatively stratify individual transcriptomic samples by clock dysfunction and globally compare clocks across individuals, conditions and tissues thus highlighting its potential relevance for advancing circadian medicine.
An efficient method for helicopter fuselage shape optimization
Authors: Zhu Jiahao , Xu Guohua , Shi Yongjie 
Published: March 2023

Purpose
This paper aims to develop a new method of fuselage drag optimization that can obtain results faster than the conventional methods based on full computational fluid dynamics (CFD) calculations and can be used to improve the efficiency of preliminary design.

Design/methodology/approach
An efficient method for helicopter fuselage shape optimization based on surrogate-based optimization is presented. Two numerical simulation methods are applied in different stages of optimization according to their relative advantages. The fast panel method is used to calculate the sample data to save calculation time for a large number of sample points. The initial solution is obtained by combining the Kriging surrogate model and the multi-island genetic algorithm. Then, the accuracy of the solution is determined by using the infill criteria based on CFD corrections. A parametric model of the fuselage is established by several characteristic sections and guiding curves.

Findings
It is demonstrated that this method can greatly reduce the calculation time while ensuring a high accuracy in the XH-59A helicopter example. The drag coefficient of the optimized fuselage is reduced by 13.3%. Because of the use of different calculation methods for samples, this novel method reduces the total calculation time by almost fourfold compared with full CFD calculations.

Originality/value
To the best of the authors’ knowledge, this is the first study to provide a novel method of fuselage drag optimization by combining different numerical simulation methods. Some suggestions on fuselage shape optimization are given for the XH-59A example.
Key factors in reducing cost of UAM implementation
Authors: Liberacki Adam , Dziugiel Bartosz , Woroniecka Paulina , Ginter Piotr , Stanczyk Anna Dorota , Mazur Anna Maria , Ten Thije Jens T. , Tojal Castro Marta 
Published: March 2023

Purpose
The purpose of the paper is the identification of the main factors affecting the cost of urban air mobility (UAM) based on results of ASSURED-UAM project. These factors can be found among such cost areas as investments (infrastructure, aircraft), operational, energy, end of life, delay and environmental. Once determined, they can be of great value for all UAM stakeholders, including manufacturers, urban planners and air service providers.

Design/methodology/approach
The obtained results were based on the outcomes of ASSURED-UAM project. Having the information about the magnitude of each cost category, we were able to identify the most costly factors of UAM. As a result, it was possible to suggest feasible cost reduction means.

Findings
For each cost category, there is the possibility to lower its value among the total cost of UAM. Each cost category has its own cost reduction means. It is vital however that the obtained results depend strongly on the assumptions made at the beginning of cost calculations.

Originality/value
The value of this paper is the identification of key UAM costs reduction means which may be found beneficial for all UAM stakeholders involved in the development of UAM infrastructure and services.
Pre-covid and peri-covid analysis of employees' work values and job quality
Authors: Amankwah Majoreen Osafroadu 
Published: March 2023

PurposeThe workspace has experienced several significant changes (shift in work culture) due to the Covid-19 pandemic thereby necessitating the need for a comparative assessment of differences that exist in work values and job quality of employees pre-covid 19 and “peri”-covid 19.Design/methodology/approachThe Statistical Package for Social Sciences (SPSS) version 21 was employed to conduct a correlation and T-test analysis from the quantitative data gathered from Ghanaian public and private sector employees.FindingsThe study found differences in what employees valued pre- and peri-covid era for job security, promotional opportunities, interesting work, using skills and experience, the usefulness of work to society and flexible work. During the pre-covid era, employees perceived autonomy, personal contact and stressful jobs, whereas promotional opportunities, good collegial relations and help to others were experienced peri-covid.Research limitations/implicationsAlthough the study tracked the work values and quality of employees, the respondents (pre-covid and peri-covid) were different. Responses here captured the views of formal sector workers only. These notwithstanding, the findings are important for explaining changes (work values and job quality) that occurred due to the pandemic. Future research might use a qualitative approach to understand reasons behind these changes.Practical implicationsDue to changes in the working space and the introduction of technology, stress has been impacted positively as employees need not commute frequently to work and work seems to have been simplified. It is therefore safe for organisations to retain changes to people management due to the pandemic.Originality/valueTo the best of my knowledge, this is the first study in the Ghanaian context comparing employees' values and job quality pre- and peri-covid.
Who and what messages are more suitable for health ads: the combined influence of endorsers and message framing on visual attention and ad effectiveness
Authors: Hsu Chiung-Wen 
Published: March 2023

PurposeThe author examined effects of endorser type and message framing on visual attention and ad effectiveness in health ads, including the moderator of involvement. This paper aims to discuss this issue.Design/methodology/approachAn experiment was conducted with a 2 (celebrity vs. expert) × 2 (positive vs. negative framing) between-subject factorial design. Eye-tracking measured visual attention and a questionnaire measured ad effectiveness and product involvement.FindingsExperimental data from 78 responses showed no vampire effect in the health advertisements. Celebrity endorsement with negative message framing received more attention and had less ad recall than that with positive message framing. Negative and positive message framing attracted the same amount of attention and ad recall in the expert endorsement condition. High involvement participants paid more attention to the ad message with the expert than that with the celebrity, but ad recall was not significantly increased. Low involvement participants exhibited the same attention to the ad message with the expert and with the celebrity, but had greater recall of the ad message with the expert. Visual attention to the endorser was associated with ad attitude but not with ad recall. Ad attitude impacted behavioral intention.Originality/valueStudies examining influences of celebrity and message framing on ad effectiveness have focused on the response to advertising stimuli, not the information process. The author provides empirical evidence of the viewers' information processing of endorsers and health messages, and its relationship with ad effectiveness. The study contributes to the literature by combining endorser and message framing in health ads to promote public health communication from the information processing perspective.
Cryptocurrency adoption and continuance intention among Indians: moderating role of perceived government control
Authors: Kala Devkant , Chaubey Dhani Shanker 
Published: March 2023

Purpose
This study aims to investigate the influence of perceived government control (PGC) on cryptocurrency adoption and continuance intention among Indians through an integrated model of the extended Unified Theory of Acceptance and Use of Technology (UTAUT) with the Information System Success Model (ISSM).

Design/methodology/approach
This study examined the items of cryptocurrency adoption, continuance intention and PGC adopted from the information systems and cryptocurrency literature. The survey was administered to 391 Indians through an online questionnaire. Partial least squares structural equation modeling was used to analyze data.

Findings
Results have shown that social influence, effort expectancy and perceived trust are the major drivers for cryptocurrency adoption. All paths leading to cryptocurrency adoption were found to be significant in the hypothesized directions. The study also found that PGC moderates the relationship between adoption and continuance intention.

Originality/value
This study advances existing literature by empirically verifying the integrated UTAUT and ISSM in the context of cryptocurrency adoption for investment purposes. The findings offer crypto-developers and crypto-exchange insight into how adoption is diffusing in emerging markets. The findings provide policymakers with meaningful insights into the role of government regulations in cryptocurrency continuance intention.
Deep and organizational learning as innovation catalyzer in digital business ecosystems – a scenario analysis on the tourism destination Berlin
Authors: Schuhbert Arne , Thees Hannes , Pechlaner Harald 
Published: March 2023

PurposeThe below-average innovative capacity of the tourism sector raises the question on the potentials of digital business ecosystems (DBEs) to overcome these shortages at a destination level – especially within a smart city environment. Using the example of the German Capital Berlin, this article aims to discuss both the possibilities and inhibitors of innovative knowledge-creation by building scenarios on one specific design option: the integration of digital deep learning (DL) functionalities and traditional organizational learning (OL) processes.Design/methodology/approachUsing the qualitative GABEK-method, major characteristics of a DBE as resource-, platform- and innovation systems are analyzed toward their interactions with the construction of basic action models (as the basic building blocks of knowledge).FindingsAgainst the background of the research findings, two scenarios are discussed for future evolution of the Berlin DBE, one building on cultural emulation as a trigger for optimized DL functionalities and one following the idea of cultural engineering supported by DL functionalities. Both scenarios focus specifically on the identified systemic inhibitors of innovative capabilities.Research limitations/implicationsWhile this study highlights the potential of the GABEK method to analyze mental models, separation of explicit and latent models still remains challenging – so does the reconstruction of higher order mental models which require a combined take on interview techniques in the future.Originality/valueThe resulting scenarios innovatively combine concepts from OL theory with the concept of DBE, thus indicating possible pathways into a tourism future where the limitations of human learning capacities could be compensated through the targeted support of general artificial intelligence (AI).
Does fit perception influence career exploration in a relational context? The mediating role of career adaptability
Authors: Kou Qi , Mohd Rasdi Roziah , Muhamad Nasharudin Nurfazreen Aina , Md Rami Ahmad Aizuddin , Cao Peng , Umar Baki Nordahlia 
Published: March 2023

Purpose
This study aims to examine the relationship between contextual factors and career exploration. Career adaptability mediates the relationship between social support, sense of belonging and career exploration.

Design/methodology/approach
This quantitative study involved 291 undergraduates recruited from two universities in China. Data were analyzed using structural equation modeling.

Findings
The results indicated that social support and sense of belonging were positively related to career adaptability. Career adaptability was found to be a mediator between relational predictors and career exploration. Contrary to our hypothesis, the direct path from sense of belonging to career exploration was insignificant.

Originality/value
This study highlights the importance of career adaptability in connecting contextual indicators and career exploration in the Chinese context.
Implementation of legislative framework governing records management throughout the life cycle in the Limpopo provincial government of South Africa
Authors: Malatji Ouma , Marutha Ngoako Solomon 
Published: March 2023

Purpose
This study aims to investigate a framework for implementation of legislative framework governing records management throughout the life cycle in the Limpopo provincial government of South Africa.

Design/methodology/approach
This quantitative multi-method study used a semi-structured questionnaire, document analysis and interviews to collect data. The dominant approach was quantitative, with some support from limited qualitative data, which served to clarify the statistical data. No sampling method was applied, as the study targeted the entire population of 135 records management participants from the provincial departments with the questionnaire, while nine participants from provincial archives participated in interviews.

Findings
The study revealed that the role of the provincial archives in enforcing compliance with legislative frameworks for the proper management of records was always disrupted by a lack of key resources such as staffing, for inspecting and training records-creating departments and working resources at the departmental levels.

Research limitations/implications
The findings will contribute to the field in respect of future studies into compliance with records management legislation, and the proposed framework may be applied as a theoretical basis for, or part of, a conceptual framework.

Practical implications
The study will serve as a resource or benchmark for archivists and records management professionals in the industry, as they count amongst the policymakers who find ways of monitoring, evaluating and enforcing compliance with the legislation governing proper records management.

Social implications
The proper implementation of recommendations from this study will lead to significant improvements in the management of records with enduring value, allowing them to finally be transferred to an archival repository to serve the public interest as heritage, national memory, or resources for researchers and authors, amongst others.

Originality/value
The study proposes a framework for implementation of legislation governing records management with greater ease in the Limpopo provincial government of South Africa.
Enhancing the design of data-related privacy controls for smart home devices
Authors: Chhetri Chola , Motti Vivian Genaro 
Published: March 2023

Purpose
Past research shows that users of smart home devices (SHDs) have privacy concerns. These concerns have been validated from technical research that shows SHDs introduce a lot of privacy risks. However, there is limited research in addressing these concerns and risks. This paper aims to bridge this gap by informing the design of data-related privacy controls for SHDs.

Design/methodology/approach
In this paper, the authors follow a user-centered design approach to design data-related privacy controls from design requirements backed by literature. The authors test the design for usability and perceived information control using psychometrically validated scales. For this purpose, two variations of the prototype (MyCam1 with a listing of data-related privacy controls and MyCam2 with three privacy presets) were created and tested them in a between-subjects experimental setting. Study participants (n = 207) were recruited via Mechanical Turk and asked to use the prototype app. An online survey was distributed to the participants to measure some usability and privacy-related constructs.

Findings
Findings show that the presented prototype designs were usable and met the privacy control needs of users. The prototype design with privacy presets (MyCam2) was found to be significantly more usable than the list of privacy controls (MyCam1).

Originality
The findings of this paper are original and build on the paper presented at the International Symposium on Human Aspects of Information Security and Assurance (HAISA 2022). This paper contributes improved and usable designs of privacy controls for smart home applications.