bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fluorescence lifetime imaging of pH along the secretory pathway
Authors: Linders Peter T.A. , Ioannidis Melina , Beest Martin ter , van den Bogaart Geert 
Published: March 2021

AbstractMany cellular processes are dependent on correct pH levels, and this is especially important for the secretory pathway. Defects in pH homeostasis in distinct organelles cause a wide range of diseases, including disorders of glycosylation and lysosomal storage diseases. Ratiometric imaging of the pH-sensitive mutant of green fluorescent protein (GFP), pHLuorin, has allowed for targeted pH measurements in various organelles, but the required sequential image acquisition is intrinsically slow and therefore the temporal resolution is unsuitable to follow the rapid transit of cargo between organelles. We therefore applied fluorescence lifetime imaging microscopy (FLIM) to measure intraorganellar pH with just a single excitation wavelength. We first validated this method by confirming the pH in multiple compartments along the secretory pathway and compared the pH values obtained by the FLIM-based measurements with those obtained by conventional ratiometric imaging. Then, we analyzed the dynamic pH changes within cells treated with Bafilomycin A1, to block the vesicular ATPase, and Brefeldin A, to block ER-Golgi trafficking. Finally, we followed the pH changes of newly-synthesized molecules of the inflammatory cytokine tumor necrosis factor (TNF)-α while they were in transit from the endoplasmic reticulum via the Golgi to the plasma membrane. The toolbox we present here can be applied to measure intracellular pH with high spatial and temporal resolution, and can be used to assess organellar pH in disease models.
Conserved and context-dependent roles for Pdgfrb signaling during zebrafish vascular mural cell development
Authors: Ando Koji , Shih Yu-Huan , Ebarasi Lwaki , Grosse Ann , Portman Daneal , Chiba Ayano , Mattonet Kenny , Gerri Claudia , Stainier Didier Y.R. , Mochizuki Naoki , Fukuhara Shigetomo , Betsholtz Christer , Lawson Nathan D. 
Published: March 2021

ABSTRACTPlatelet derived growth factor beta and its receptor, Pdgfrb, play essential roles in the development of vascular mural cells, including pericytes and vascular smooth muscle. To determine if this role was conserved in zebrafish, we analyzedpdgfbandpdgfrbmutant lines. Similar to mouse,pdgfbandpdgfrbmutant zebrafish lack brain pericytes and exhibit anatomically selective loss of vascular smooth muscle coverage. Despite these defects,pdgfrbmutant zebrafish did not otherwise exhibit circulatory defects at larval stages. However, beginning at juvenile stages, we observed severe cranial hemorrhage and vessel dilation associated with loss of pericytes and vascular smooth muscle cells inpdgfrbmutants. Similar to mouse,pdgfrbmutant zebrafish also displayed structural defects in the glomerulus, but normal development of hepatic stellate cells. We also noted defective mural cell investment on coronary vessels with concomitant defects in their development. Together, our studies support a conserved requirement for Pdgfrb signaling in mural cells. In addition, these mutants provide an important model for definitive investigation of mural cells during early embryonic stages without confounding secondary effects from circulatory defects.Summary statementGenetic analysis in zebrafish demonstrates the conserved role of Pdgfb/Pdgfrb signaling in pericyte and vascular smooth muscle cell formation during vascular development in vertebrates.
A MALDI-TOF assay identifies nilotinib as an inhibitor of inflammation in acute myeloid leukaemia
Authors: Marín-Rubio José Luis , Heap Rachel E. , Heunis Tiaan , Dueñas Maria Emilia , Inns Joseph , Scott Jonathan , Simpson A. John , Blair Helen , Heidenreich Olaf , Allan James M. , Saxty Barbara , Trost Matthias 
Published: March 2021

ABSTRACTInflammatory responses are important in cancer, particularly in the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs in human monocytes derived from acute myeloid leukaemia (AML), by tracking several biological features ionizing from only 2,500 cells using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. A proof-of-concept screen showed that the BCR-ABL inhibitor nilotinib, but not the structurally similar imatinib, blocks inflammatory responses. In order to identify the cellular (off-)targets of nilotinib, we performed thermal proteome profiling (TPP). Unlike imatinib, nilotinib and other later generation BCR-ABL inhibitors inhibit the p38α-MK2/3 signalling axis which suppressed the expression of inflammatory cytokines, cell adhesion and innate immunity markers in activated human monocytes derived from AML. Thus, our study provides a tool for the discovery of new anti-inflammatory drugs, which could contribute to the treatment of inflammation in myeloid neoplasms and other diseases.Key PointsLabel-free cell-based assay identifies new anti-inflammatory drugs using MALDI-TOF MS. Nilotinib reduces inflammation by inhibition of MAPK14-MK2/3 signalling axis in AML.
Long-read cDNA sequencing identifies functional pseudogenes in the human transcriptome
Authors: Troskie Robin-Lee , Jafrani Yohaann , Mercer Tim R. , Ewing Adam D. , Faulkner Geoffrey J. , Cheetham Seth W. 
Published: March 2021

AbstractPseudogenes are gene copies presumed to mainly be functionless relics of evolution due to acquired deleterious mutations or transcriptional silencing. When transcribed, pseudogenes may encode proteins or enact RNA-intrinsic regulatory mechanisms. However, the extent, characteristics and functional relevance of the human pseudogene transcriptome are unclear. Short-read sequencing platforms have limited power to resolve and accurately quantify pseudogene transcripts owing to the high sequence similarity of pseudogenes and their parent genes. Using deep full-length PacBio cDNA sequencing of normal human tissues and cancer cell lines, we identify here hundreds of novel transcribed pseudogenes. Pseudogene transcripts are expressed in tissue-specific patterns, exhibit complex splicing patterns and contribute to the coding sequences of known genes. We survey pseudogene transcripts encoding intact open reading frames (ORFs), representing potential unannotated protein-coding genes, and demonstrate their efficient translation in cultured cells. To assess the impact of noncoding pseudogenes on the cellular transcriptome, we delete the nucleus-enriched pseudogene PDCL3P4 transcript from HAP1 cells and observe hundreds of perturbed genes. This study highlights pseudogenes as a complex and dynamic component of the transcriptional landscape underpinning human biology and disease.
The DBL-1/TGF-β signaling pathway regulates pathogen-specific innate immune responses inC. elegans
Authors: Madhu Bhoomi , Gumienny Tina L. 
Published: March 2021

ABSTRACTInnate immunity in animals is orchestrated by multiple cell signaling pathways, including the TGF-β superfamily pathway. While the role of TGF-β signaling in innate immunity has been clearly identified, the requirement for this pathway in generating specific, robust responses to different bacterial challenges has not been characterized. Here, we address the role of DBL-1/TGF-β in regulating signature host defense responses to a wide range of bacteria inC. elegans. This work reveals a role of DBL-1/TGF-β in animal survival, organismal behaviors, and molecular responses in different environments. Additionally, we identify a novel role for SMA-4/Smad that suggests both DBL-1/TGF-β-dependent and -independent functions in host avoidance responses. RNA-seq analyses and immunity reporter studies indicate DBL-1/TGF-β differentially regulates target gene expression upon exposure to different bacteria. Furthermore, the DBL-1/TGF-β pathway is itself differentially affected by the bacteria exposure. Collectively, these findings demonstrate bacteria-specific host immune responses regulated by the DBL-1/TGF-β signaling pathway.
Pre-service teachers' acquisition of scientific knowledge and scientific skills through inquiry-based laboratory activity
Authors: Valls-Bautista Cristina , Solé-LLussà Anna , Casanoves Marina 
Published: March 2021

PurposeScientific inquiry is a leading methodology that promotes science process skills to acquire scientific knowledge. There is evidence that primary school teachers have difficulties introducing inquiry-based activities in their classrooms. Hence, adequate teacher instruction in inquiry methodology is important to apply inquiry-based activities in school science lessons. This work aims to analyse if pre-service teachers succeeded in developing scientific knowledge and scientific skills through the application of an inquiry laboratory activity.Design/methodology/approachThis article is presented as a case study developed in a group of 82 pre-service teachers. This research methodology involved qualitative and quantitative data.FindingsThe results demonstrate that pre-service teachers could improve their scientific skills and knowledge through inquiry-based laboratory activity.Originality/valueThe present study assesses not only the scientific knowledge but also if students can acquire scientific skills by doing the inquiry laboratory activity and if these skills are related to low-order cognitive skills or high-order cognitive skills.
Five leading-edge K-12 districts’ decision-making processes for EdTech innovations
Authors: Dexter Sara , Francisco Aubrey , Luke Luna Christina 
Published: March 2021

PurposeThe purpose of this study was to better understand K-12 district leaders' reasoning and processes for selecting and deploying EdTech instructional products, including which, if any, types of data are used to support decision-making.Design/methodology/approachThis multisite case study of educational technology (EdTech) decision-making comprises five purposely selected districts at the leading edge of EdTech innovation. The unit of analysis was a recent purchase they had made of an instructional, classroom-oriented digital product (defined as a product used by teachers and/or students in the classroom for the purposes of student learning). The key leader heading up the purchase was interviewed, as were other leaders and a teacher who were involved in the decision-making process.FindingsThe processes districts used to make their purchasing decisions involved teachers, district leaders and technical specialists who considered usability, usage data and alignment with student learning and interoperability, respectively. While in some cases there were plans to collect data on student learning outcomes, districts did not uniformly emphasize that in their decision-making processes. Instead, the type of educational technology tool that was purchased influenced whether or not districts planned to seek out student-level outcome data as evidence of the product's efficacy. For the purchases associated with access to content, school leaders considered usage or log data generated by the program itself as sufficient indication that the program is “working.” Where the software's functionality encompassed skill development, leaders stated future plans to look at student-level outcomes as a means for judging if the program “worked.”Originality/valueFew accounts of district decision-making about the adoption of educational technology innovations are present in the literature. These five cases provide insight into the role evidence plays in decisions to adopt educational technology.
Examining year-end spending spikes in the European Economic Area: a comparative study of procurement contracts
Authors: McCue Clifford P. , Prier Eric , Lofaro Ryan J. 
Published: March 2021

PurposeThe purpose of this study is to analyze year-end spending practices in the European Economic Area (EEA) to baseline the pervasiveness of year-end spending spikes across countries in Europe.Design/methodology/approachThe Tenders Electronic Daily dataset is used to descriptively analyze above-threshold procurement contracts by country, year and contract type from 2009 to 2018. Proportional distributions are employed to compare percentages of spend across quarters. Analyses are run within each country on the number of years displaying a fourth quarter spike, as well as within each country and contract type.FindingsThe results show that while spending spikes for above-threshold contracts in the final fiscal quarter are not consistent across all countries, patterns emerge when the data are disaggregated by country. The most populous nations in the EEA are more likely to have years with the highest proportion of fiscal spend occurring in the fourth quarter. Further, the type of contract makes a difference – services and supplies contracts are more likely to display fourth quarter spikes than works contracts.Originality/valueThis article provides the first analysis of the year-end spending spike across countries in Europe using procurement data, as well as the first to disaggregate by year and contract type. Findings support the literature on the presence of year-end spikes; such spikes exist even for above-threshold public procurement contracts.
How user behaviour is moderated by affective commitment on point of sale terminal
Authors: Oloveze Ambrose Ogbonna , Oteh Ogbonnaya Ukeh , Nwosu Hyginus Emeka , Obasi Ray Ozoemena 
Published: March 2021

PurposeSeveral e-payment technologies have diffused in Nigeria yet debit card usage on POS devices have not shown consumer confidence in its usage thereby affecting the cashless policy drive of the nation. The study considered an affective commitment of users as moderating their behaviours. Therefore, the purpose of this paper is to investigate how user behaviour is moderated by an affective commitment on point of sale terminal.Design/methodology/approachFollowing the purpose of the study, the research design is a survey. The questionnaire was adapted from earlier studies and tested for reliability and validity using Cronbach’s alpha and content validity. Andrew F. Hayes process was used to analyse the moderation effect.FindingsThe finding revealed that affective commitment significantly moderates users’ ease of use of the device, their perceived usefulness of the device and their social image on their intentions to use the device.Research limitations/implicationsThe findings implied that there is a need for the development of policies and strategies which should be directed towards the users of the device. Equally, the general conclusions on collective e-payment channels in a society should be discarded given that each e-payment channel can have different factors influencing it than the others. This is where customer-focused advertising and awareness campaign becomes very important.Originality/valueThis paper declares that the research work is not submitted anywhere for publication or for review. It was conducted by the authors who have given their consent for it to be submitted to Rajagiri management journal
Expression of CD1 molecules and colony‐stimulating factor 1 receptor in indeterminate cell histiocytosis
Authors: Kinoshita Manao , Ogawa Youichi , Honobe Akiko , Tomita Ozumi , Oishi Naoki , Shimada Shinji , Kondo Tetsuo , Kawamura Tatsuyoshi 
Published: March 2021

AbstractIndeterminate cell histiocytosis (ICH) and Langerhans cell histiocytosis (LCH) are rare histiocyte proliferating disorders with unknown etiologies. However, both tumor cells immunophenotypically share some features of Langerhans cells (LC), thereby expressing CD1a. Recent transcriptome analysis revealed that circulating CD1c+ myeloid dendritic cells are the potential precursor of LCH tumor cells. LC express CD1a as well as CD1c, but not CD1b. We discovered that both tumor cells express CD1c, but not CD1b, similar to LC. Moreover, like LC, both tumor cells express colony‐stimulating factor 1 receptor (CSF‐1R). Considering the crucial role of the interleukin (IL)‐34/CSF‐1R axis for the development and survival of LC, CSF‐1R on both tumor cells might facilitate their survival and proliferation in situ. These data provide additional evidence to support the fact that ICH and LCH share immunophenotypical features with LC. In addition, we hypothesized that tumor cells in ICH and LCH survive and proliferate through IL‐34‐mediated CSF‐1R signaling.
Anonymous Equity Research
Authors: DYER TRAVIS , KIM EUNJEE 
Published: March 2021

ABSTRACTCrowdsourced financial information platforms often allow content contributors to publish equity research anonymously. This study examines whether investors value or discount information in anonymous equity research. In the short window around research releases, we find that investors’ stock price reaction to anonymous research is muted in comparison to nonanonymous research. Consistent with credibility concerns influencing investor response, we document that this discount to anonymous research dissipates as the monitoring of content contributors intensifies and as authors develop a reputation for high‐quality reporting. In addition, we perform a content analysis on the research reports and find that the muted market reaction to anonymous equity research is robust to controlling for textual attributes of information content, further supporting our inference that investors’ are concerned about the credibility of anonymous equity research.
The effect of respiratory activity, non‐invasive respiratory support and facemasks on aerosol generation and its relevance to COVID‐19
Authors: Wilson N. M. , Marks G. B. , Eckhardt A. , Clarke A. M. , Young F. P. , Garden F. L. , Stewart W. , Cook T. M. , Tovey E. R. 
Published: March 2021

SummaryRespirable aerosols (< 5 µm in diameter) present a high risk of SARS‐CoV‐2 transmission. Guidelines recommend using aerosol precautions during aerosol‐generating procedures, and droplet (> 5 µm) precautions at other times. However, emerging evidence indicates respiratory activities may be a more important source of aerosols than clinical procedures such as tracheal intubation. We aimed to measure the size, total number and volume of all human aerosols exhaled during respiratory activities and therapies. We used a novel chamber with an optical particle counter sampling at 100 l.min‐1 to count and size‐fractionate close to all exhaled particles (0.5–25 µm). We compared emissions from ten healthy subjects during six respiratory activities (quiet breathing; talking; shouting; forced expiratory manoeuvres; exercise; and coughing) with three respiratory therapies (high‐flow nasal oxygen and single or dual circuit non‐invasive positive pressure ventilation). Activities were repeated while wearing facemasks. When compared with quiet breathing, exertional respiratory activities increased particle counts 34.6‐fold during talking and 370.8‐fold during coughing (p < 0.001). High‐flow nasal oxygen 60 at l.min‐1 increased particle counts 2.3‐fold (p = 0.031) during quiet breathing. Single and dual circuit non‐invasive respiratory therapy at 25/10 cm.H2O with quiet breathing increased counts by 2.6‐fold and 7.8‐fold, respectively (both p < 0.001). During exertional activities, respiratory therapies and facemasks reduced emissions compared with activities alone. Respiratory activities (including exertional breathing and coughing) which mimic respiratory patterns during illness generate substantially more aerosols than non‐invasive respiratory therapies, which conversely can reduce total emissions. We argue the risk of aerosol exposure is underappreciated and warrants widespread, targeted interventions.
Questioning the utility of round spermatid injections in men with non‐obstructive azoospermia
Authors: Barda Shimi , Mano Roy , Lehavi Ofer , Kleiman Sandra E. , Yossepowitch Ofer , Azem Foad , Hauser Ron , Dekalo Snir 
Published: March 2021

AbstractBackgroundData on who among the infertile male population may benefit from round spermatid injections (ROSI) are lacking.ObjectiveTo determine the probability of finding round spermatids suitable for ROSI in men with non‐obstructive azoospermia (NOA) in whom no spermatozoa were retrieved at testicular sperm extraction.Materials and methodsFour‐hundred fifty‐seven consecutive men with azoospermia underwent testicular sperm extraction. Clinical examination included age, secondary sexual characteristics, testicular size, reproductive hormone estimation, karyotyping, and Y chromosome microdeletion analyses. Histologic examination was performed, and histologic classification was determined by the most advanced spermatogenetic cell identified in the combined histologic and cytologic examination.ResultsOf the 457 azoospermic men, 342 were diagnosed with NOA, and 148 (148/342, 43%) had mixed atrophy on histopathology and retrievable spermatozoa. No spermatozoa were found in 194/342 men with NOA (57%). Histopathology diagnosed 145/194 (75%) of them with Sertoli cell only, 45/194 (23%) with spermatocyte maturation arrest, and 4/194 (2%) with spermatid maturation arrest.ConclusionsHistopathologically identified round spermatids without spermatozoa were rare in men with NOA. Only very few of them are likely to reap the benefits of ROSI, thus presenting the need to reconsider its actual clinical value.
Advances in understanding the pathogenesis of hereditary macrothrombocytopenia
Authors: Collins Janine , Astle William J. , Megy Karyn , Mumford Andrew D. , Vuckovic Dragana 
Published: March 2021

SummaryLow platelet count, or thrombocytopenia, is a common haematological abnormality, with a wide differential diagnosis, which may represent a clinically significant underlying pathology. Macrothrombocytopenia, the presence of large platelets in combination with thrombocytopenia, can be acquired or hereditary and indicative of a complex disorder. In this review, we discuss the interpretation of platelet count and volume measured by automated haematology analysers and highlight some important technical considerations relevant to the analysis of blood samples with macrothrombocytopenia. We review how large cohorts, such as the UK Biobank and INTERVAL studies, have enabled an accurate description of the distribution and co‐variation of platelet parameters in adult populations. We discuss how genome‐wide association studies have identified hundreds of genetic associations with platelet count and mean platelet volume, which in aggregate can explain large fractions of phenotypic variance, consistent with a complex genetic architecture and polygenic inheritance. Finally, we describe the large genetic diagnostic and discovery programmes, which, simultaneously to genome‐wide association studies, have expanded the repertoire of genes and variants associated with extreme platelet phenotypes. These have advanced our understanding of the pathogenesis of hereditary macrothrombocytopenia and support a future clinical diagnostic strategy that utilises genotype alongside clinical and laboratory phenotype data.