bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol
Authors: Loll-Krippleber Raphael , Sajtovich Victoria , Ferguson Michael W. , Ho Brandon , Payliss Brandon J. , Bellissimo Joseph , Peters Sydney , Wyatt Haley D. M. , Brown Grant W. 
Published: October 2022

AbstractBackgroundOver the 70 years since the introduction of plastic into everyday items, plastic waste has become an increasing problem. With over 360 million tonnes of plastics produced every year, solutions for plastic recycling and plastic waste reduction are sorely needed. Recently, multiple enzymes capable of degrading PET (polyethylene teraphthalate) plastic have been identified and engineered. In particular, the enzymes PETase and MHETase fromIdeonella sakaiensisdepolymerize PET into the two building blocks used for its synthesis, ethylene glycol (EG) and terephthalic acid (TPA). Importantly, EG and TPA can be re-used for PET synthesis allowing complete and sustainable PET recycling.ResultsIn this study, we usedSaccharomyces cerevisiaeas a platform to develop a whole-cell catalyst expressing the MHETase enzyme, which converts MHET (monohydroxyethyl terephthalate) into TPA and EG. We assessed six expression architectures and identified those resulting in efficient MHETase expression on the yeast cell surface. We show that the MHETase whole-cell catalyst has activity comparable to recombinant MHETase purified fromEscherichia coli. Finally, we demonstrate that surface displayed MHETase is stable to pH, temperature, and for at least 12 days at room temperature.ConclusionsWe demonstrate the feasibility of usingS. cerevisiaeas a platform for the expression and surface display of PET degrading enzymes and predict that the whole-cell catalyst will viable alternatives to protein purification-based approaches for plastic degradation.
Understanding the necessity of regulatory protein machinery in heterologous expression of class-III type of Ocins
Authors: Choyam Shilja , Kammara Rajagopal. 
Published: October 2022

AbstractTo date, there have been no or just a few reports of successful cloning and expression to create biologically active ocins or bacteriocins. Cloning, expression, and production of class I ocins are problematic because of their structural arrangements, coordinated functions, size, and posttranslational modifications. In the case of class III ocins, there are no reports of obtaining biological active proteins to date. Because of their growing importance, use and rapid functions require understanding mechanistic aspects to obtain biological active protein. As a result, we intend to clone and express the class III type. Also, by fusion or chimaera, they reshape class I types that lack posttranslational modifications into class III. Therefore, this construct resembles a class III type ocin. With the exception of Zoocin, expression of the proteins was found to be physiologically ineffective after cloning. But, few cell morphological changes such as elongation, aggregation, and the formation of terminal hyphae were observed. However, it was discovered that the target indicator had been altered toVibrio spp. in a few. Finally, we confirm the existence of unidentified additional intrinsic factors for succesful expression to obtain biologically active protein.
An Optimized Electrotransformation Protocol forLactobacillus jensenii
Authors: Fristot Elsa , Bessede Thomas , Rufino Miguel Camacho , Mayonove Pauline , Chang Hung-Ju , Zuniga Ana , Michon Anne-Laure , Godreuil Sylvain , Bonnet Jérôme , Cambray Guillaume 
Published: October 2022

ABSTRACTEngineered bacteria are promising candidates forin situdetection and treatment of diseases. The female uro-genital tract presents several pathologies, such as sexually transmitted diseases or genital cancer, that could benefit from such technology. While bacteria from the gut microbiome are increasingly engineered, the use of chassis isolated from the female uro-genital resident flora has been limited. A major hurdle to implement the experimental throughput required for efficient engineering in these non-model bacteria is their low transformability. Here we report an optimized electrotransformation protocol forLactobacillus jensenii, one the most widespread species across vaginal microflora. Starting from classical conditions, we optimized buffers, electric field parameters, cuvette type and DNA quantity to achieve an 80-fold improvement in transformation efficiency, with up to 3.5·103CFUs/µg of DNA inL. jensenii ATCC 25258. We also identify several plasmids that are maintained and support reporter gene expression inL. jensenii. Finally, we demonstrate that our protocol provides increased transformability in three independent clinical isolates ofL. jensenii. This work will facilitate the genetic engineering ofL. jenseniiand enable its use for addressing challenges in gynecological healthcare.
SMDT1variants impair EMRE-mediated mitochondrial calcium uptake in patients with muscle involvement
Authors: Bulthuis Elianne P. , Adjobo-Hermans Merel J.W. , de Potter Bastiaan , Hoogstraten Saskia , Wezendonk Lisanne H.T. , Tutakhel Omar A.Z. , Wintjes Liesbeth T. , van den Heuvel Bert , Willems Peter H.G.M. , Kamsteeg Erik-Jan , Rubio Gozalbo M. Estela , Sallevelt Suzanne C.E.H. , Koudijs Suzanne M. , Nicolai Joost , de Bie Charlotte I. , Hoogendijk Jessica E. , Koopman Werner J.H. , Rodenburg Richard J. 
Published: October 2022

AbstractIonic calcium (Ca2+) is a key messenger in signal transduction and its mitochondrial uptake plays an important role in cell physiology. This uptake is mediated by the mitochondrial Ca2+uniporter (MCU), which is regulated by EMRE (essential MCU regulator) encoded by theSMDT1(single-pass membrane protein with aspartate rich tail 1) gene. This work presents the genetic, clinical and cellular characterization of two patients harbouringSMDT1variants and presenting with muscle problems. Analysis of patient fibroblasts and complementation experiments provide evidence that these variants lead to absence of EMRE protein, induce MCU subcomplex formation and impair mitochondrial Ca2+uptake. However, the activity of the oxidative phosphorylation enzymes, mitochondrial morphology and membrane potential, as well as routine/ATP-linked respiration were not affected. We hypothesize that the muscle-related symptoms in the patients withSMDT1variants result from aberrant mitochondrial Ca2+uptake.
The molecular principles governing HCMV infection outcome
Authors: Schwartz Michal , Shnayder Miri , Nachshon Aharon , Arazi Tamar , Kitsberg Yaarit , Samia Roi Levi , Lavi Michael , Kuint Rottem , Tsabari Reuven , Stern-Ginossar Noam 
Published: October 2022

AbstractInfection with Human cytomegalovirus (HCMV) can result in either productive or non-productive infection, the latter potentially leading to establishment of latency, but the molecular factors that dictate these different infection outcomes are elusive. Macrophages are known targets of HCMV and considered to be permissive for productive infection, while monocytes, their precursors, are latently infected. Here we reveal that infection of macrophages is more complex than previously appreciated and can result in either productive or non-productive infection. By analyzing the progression of HCMV infection in monocytes and macrophages using single cell transcriptomics, we uncover that the level of viral gene expression, and specifically the expression of the major immediate early proteins, IE1 and IE2, is the principal barrier for establishing productive infection. On the cellular side, we reveal that the cell intrinsic levels of interferon stimulated genes (ISG), but not their induction, is a main determinant of infection outcome and that intrinsic ISG levels are downregulated with monocyte differentiation, partially explaining why macrophages are more susceptible to productive HCMV infection. We further show that, compared to monocytes, non-productive macrophages maintain higher levels of viral transcripts and are able to reactivate, raising the possibility that they may serve as latency reservoirs. Overall, by harnessing the tractable system of monocyte differentiation we decipher underlying principles that control HCMV infection outcome, and propose macrophages as a potential HCMV reservoir in tissues.
Genomic Sequencing from Sputum for Tuberculosis Disease Diagnosis, Lineage Determination and Drug Susceptibility Prediction
Authors: Nilgiriwala Kayzad , Rabodoarivelo Marie-Sylvianne , Hall Michael B. , Patel Grishma , Mandal Ayan , Mishra Shefali , Andrianomanana Fanantenana Randria , Dingle Kate , Rodger Gillian , George Sophie , Crook Derrick W. , Hoosdally Sarah , Mistry Nerges , Rakotosamimanana Niaina , Iqbal Zamin , Lapierre Simon Grandjean , Walker Timothy M. 
Published: October 2022

AbstractBackgroundUniversal access to drug susceptibility testing for newly diagnosed tuberculosis patients is recommended. Access to culture-based diagnostics remains limited and targeted molecular assays are vulnerable to emerging resistance conferring mutations. Improved sample preparation protocols for direct-from-sputum sequencing ofMycobacterium tuberculosiswould accelerate access to comprehensive drug susceptibility testing and molecular typing.MethodsWe assessed a thermo-protection buffer-based direct-from-sampleM. tuberculosiswhole-genome sequencing protocol. We prospectively processed and analyzed 60 acid-fast bacilli smear-positive sputum samples from tuberculosis patients in India and Madagascar. A diversity of semi-quantitative smear positivity level samples were included. Sequencing was performed using Illumina and MinION (monoplex and multiplex) technologies. We measured the impact of bacterial inoculum and sequencing platforms onM. tuberculosisgenomic mean read depth, drug susceptibility prediction performance and typing accuracy.ResultsM. tuberculosiswas identified from 88% (Illumina), 89% (MinION-monoplex) and 83% (MinION-multiplex) of samples for which sufficient DNA could be extracted. The fraction ofM. tuberculosisreads from MinION sequencing was lower than from Illumina, but monoplexing grade 3+ sputum samples on MinION produced higher read depth than Illumina (p<0.05) and MinION multiplex (p<0.01). No significant difference in overall sensitivity and specificity of drug susceptibility predictions was seen across these sequencing modalities or within each sequencing technology when stratified by smear grade. Lineage typing agreement percentages between direct and culture-based sequencing were 85% (MinION-monoplex), 88% (Illumina) and 100% (MinION-multiplex)ConclusionsM. tuberculosisdirect-from-sample whole-genome sequencing remains challenging. Improved and affordable sample treatment protocols are needed prior to clinical deployment.
Recruiting volunteers: the roles of fear, hope and courage
Authors: Chowdhury Rafi M.M.I. , Septianto Felix 
Published: October 2022

Purpose
Nonprofit organizations face challenges recruiting volunteers for morally important activities that may generate fear, such as firefighting, aid work and delinquent counseling. The purpose of this study is to examine how voluntary organizations can instill the virtue of courage among potential volunteers and motivate them to participate in such activities.

Design/methodology/approach
Three experimental studies examined how fear, hope and courage relate to the likelihood of volunteering. Study 1 investigated how integral hope (hope related to the context, i.e. hope emanating from volunteering activities) and incidental hope (hope unrelated to the context, i.e. a general hopeful feeling) affect volunteering intentions when there is low vs high fear. Study 2 examined whether courage mediated the effects of hope on volunteering intentions when there is low vs high fear. Study 3 replicated the findings in a different volunteering context.

Findings
Integral hope (but not incidental hope) in the face of high fear generates courage leading to intentions to volunteer. Both integral hope and incidental hope motivate volunteering intentions through positive affect (but not through courage) in low fear contexts.

Research limitations/implications
The hypothetical volunteering scenarios and the gender distribution in the samples restrict the external validity of the findings. Family background in volunteering was not controlled for. Moral courage, physical courage and psychological courage were not separately measured.

Practical implications
Nonprofit organizations recruiting volunteers for risky voluntary activities that induce high fear should use integral hope in their marketing communications to instill courage among potential volunteers. For voluntary activities that are not very risky and generate low levels of fear among potential volunteers, nonprofit organizations can recruit volunteers through communications that use either integral hope or incidental hope.

Originality/value
This research shows that hope and fear are critical emotions in relation to courage – an essential virtue for volunteers. Courage is manifested when there is high fear and integral hope. Findings contribute to the research literatures on the marketing of volunteering and the moral psychology of courage.
Dividend and investment decisions in asymmetric information environments: evidence from the restaurant industry
Authors: Gim Jaehee , Jang SooCheong (Shawn) 
Published: October 2022

Purpose
This study aims to examine how information asymmetry, which refers to an information gap between a firm’s management and its investors regarding the firm’s true value, influences firms’ dividend and investment decisions in the restaurant industry. This study also investigated the moderating role of a firm’s level of franchising in the relationship between information asymmetry and these behaviors of restaurant firms.

Design/methodology/approach
This study used generalized method of moments panel regression analyses. Principal component analysis was also used to create a composite index of information symmetry.

Findings
This study demonstrated that in asymmetric information environments, restaurant managers tend to reduce dividend payments. In addition, this study showed that information asymmetry leads to restaurant managers’ investment inefficiency. However, the investment inefficiency of the restaurant industry was found to decrease as restaurant firms’ level of franchising increases.

Practical implications
Firms’ dividends and investment decisions are of great interest to investors because these decisions heavily influence investors’ wealth-maximization goals. By shedding light on the previously unrecognized determinants of dividend and investment behaviors in the restaurant industry, this study helps individual investors to make informed investing decisions.

Originality/value
Conflicting arguments can be made regarding the impact of asymmetric information environments on the dividend and investment behaviors of restaurant firms. This study aimed to verify these as-yet unclear relationships in the restaurant industry.
Developing structural modelling of inter-partner factors of international joint ventures performance
Authors: Batra Ishita , Dhir Sanjay 
Published: October 2022

PurposeThe study aims to identify, analyse and develop a model for measuring the inter-relationship and interaction among the inter-partner factors. International joint ventures (IJVs) literature has scantly studied the interactions and inter-relationships among the inter-partner factors of IJVs performance. To address this gap, this research creates a hierarchical relationship framework among the inter-partner factors of IJVs performance.Design/methodology/approachThe comprehensive literature review is used to identify the factors and possible relationships between the factors and IJVs performance. M-TISM (modified total interpretive structural modelling) methodology is used to examine the relationship among the factors of IJVs performance.FindingsThis study highlights 12 inter-partner factors that affect IJVs performance. The results suggest that size asymmetry, commitment, goal incongruency, competitive overlap, trust, control, the interdependency of resources, cooperation, economic distance, cultural distance, geographical distance and administrative distance significantly impact IJVs performance. Commitment is found to be a linkage variable as it has high dependence and driving power. Goal incongruency and competitive overlap have high driving power.Research limitations/implicationsThis study gives a proposed list of critical inter-partner factors for policymakers and practitioners to consider when developing rules or recommendations to improve IJVs performance. The focus of policymakers should be on economic and geographical distance, followed by cultural distance, competitive overlap, goal incongruency, size asymmetry and administrative distance to improve inter-relationship among the partners in IJVs. The work contributes to the academician by presenting a modified TISM model that highlights the significance of ambidexterity in driving the performance of IJVs.Originality/valueThis study fills a gap in the literature by analysing the interactions among the inter-partner parameters that influence the performance of IJVs. It offers insights into the variables, such as driver dependence and the relationships between the variables.
Firm characteristics and forward-looking disclosure: the moderating role of gender diversity
Authors: Abdelazim Samir Ibrahim , Metwally Abdelmoneim Bahyeldin Mohamed , Aly Saleh Aly Saleh 
Published: October 2022

PurposeThe purpose of this study is to examine the impact of firm financial and operational characteristics on the level of forward-looking information disclosure (FLID) by Egyptian-listed non-financial companies. The present research also aims to investigate the moderating role of gender diversity on the board of directors.Design/methodology/approachThe sample incorporates the non-financial companies included in the EGX 100 of the Egyptian Stock Exchange (ESE), whose reports were available during the study period from 2013 to 2018. The final sample comprises 49 companies with 294 observations. Statistical analysis is performed using multiple regression analysis.FindingsThis study found a significant positive impact of return on assets, leverage, company size and age on the level FLID, while external audit firm type and industry were found to impact the level of FLID negatively. Further, the board gender diversity (BGD) is found to have a moderating impact as it strengthens the effect of financial and operational characteristics on the level of FLID.Practical implicationsThe present study has some implications for Egyptian companies, investors in the Egyptian market and regulators in emerging economies, which include paying more attention to BGD when selecting the board members by companies as well as following up the female representation in all the listed companies by regulators.Originality/valueTo the best of the authors’ knowledge, this is the first study to investigate the moderating role of BGD and its impact on the level of FLID in emerging markets. This extends the disclosure literature as the present study brings new evidence from an emerging market regarding BGD moderating role as early research concentrated on the direct impact of BGD on the level of FLID.
Risk-taking and WOM as moderators in the relationship between status consumption, brand image and purchase intention of counterfeit brand shoes
Authors: Patel Ashok Kumar , Singh Anurag , Parayitam Satyanarayana 
Published: October 2022

PurposeThe study's objective is to examine the consumers' intention to buy counterfeit brand shoes. A conceptual model is developed to test the risk-taking and word-of-mouth (WOM) as a moderator in the relationship between status consumption, brand image, and consumer intention to buy counterfeit shoes.Design/methodology/approachBased on the theory of reasoned action (TRA) and signaling theory (ST), this research was conducted in the Indian National Capital Region. Using a structured instrument, the data was collected from 240 respondents. After checking the psychometric properties of the survey instrument using the Lisrel package of structural equation modeling, Hayes's PROCESS macros were used for testing the hypotheses.FindingsThe findings from the study indicate that (1) status consumption and brand image are positively associated with purchase intention of counterfeit brand shoes, and (2) risk-taking moderates the relationship between (1) status consumption and purchase intention, and (2) brand image and purchase intension, (3) significant three-way interaction between WOM, risk-taking and status consumption on purchase intention, and (4) significant three-way interaction between brand image, WOM, and risk-taking on purchase intention of counterfeit brand shoes.Research limitations/implicationsAs with any survey research, this study has common method variance as a potential problem. However, through the latent variable method and Harman's single-factor analysis, the common method variance was checked. The study has several implications for managers, e-marketers, and consumers.Practical implicationsThe study has several implications for marketers selling counterfeit products and managers intending to protect their branded products.Originality/valueA conceptual model showing two-way and three-way interactions between status consumption, risk-taking, and WOM influencing the consumer purchase intention of counterfeit products was discussed. This is the first of its kind in India to explore such relationships.
Impact of the time limits for the receipt of tenders on the number of bidders: evidence from public procurement in Greece
Authors: Pliatsidis Andreas Christos 
Published: October 2022

PurposeThe purpose of this paper is to examine how the period for which a public procurement notice remains open for bidding (Δt) affects the number of bids.Design/methodology/approachThe authors investigated data for 2.404 open procurement tenders in Greece for the years 2018–2021. Using Δtas the grouping factor, the authors defined two samples based on the European Union time limits for the receipt of tenders. Group 1 (Δt≤ 35) contains all tenders for which the contracting authorities (CAs) have chosen to limit themselves to the minimum number of days allowed by law. Group 2 (Δt>35) includes the remaining tenders where CAs have chosen to keep their notices open for periods beyond the existing minimum time limits, as they are encouraged to do by law.FindingsA Mann–WhitneyUtest, in combination with graphical analysis, revealed that CAs from Group 2 tend to enjoy more bids per tender, that is, more intense competition.Social implicationsThe paper allows decision-makers and legislators to understand the relationship between the time CAs choose to keep their notices open for bidding and the number of bidders in each tender, that is, competition, which according to other authors, affects the outcomes of public procurement procedures.Originality/valueThe paper fills the research gap regarding the relationship between time for preparation and the number of bids in each tender.