bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression and purification of human neutrophil proteinase 3 from insect cells and characterization of ligand binding
Authors: Khorsand Fahimeh , Haug Bengt Erik , Kursula Inari , Reuter Nathalie , Brenk Ruth 
Published: November 2023

AbstractNeutrophil proteinase 3 (PR3) is an important drug target for inflammatory lung diseases such as chronic obstructive pulmonary disease and cystic fibrosis. Drug discovery efforts targeting PR3 require active enzyme forin vitrocharacterization, such as inhibitor screening, enzymatic assays, and structural studies. Recombinant expression of active PR3 overcomes the need for enzyme supplies from human blood and in addition allows studies on the influence of mutations on enzyme activity and ligand binding. Here, we report the expression of recombinant PR3 (rPR3) using a baculovirus expression system. The purification and activation process described resulted in highly pure and active PR3. The activity of rPR3 in the presence of commercially available inhibitors was compared with human PR3 by using a fluorescence-based enzymatic assay. Purified rPR3 had comparable activity to the native human enzyme, thus being a suitable alternative for enzymatic studiesin vitro. Further, we established a surface plasmon resonance-based assay to determine binding affinities and kinetics of PR3 ligands. These methods provide valuable tools for early drug discovery aiming towards treatment of lung inflammation.
MiR-221/222-enriched ADSC-exosome mitigates PM exposure-exacerbated cardiac ischemia/reperfusion injury through the modulation of the BNIP3/LC3B/PUMA pathway
Authors: Lee Tzu-Lin , Shen Wen-Chi , Chen Ya-Chun , Lai Tsai-Chun , Lin Shu-Rung , Lin Shu-Wha , Yu I-Shing , Yeh Yen-Hsiu , Li Tsai-Kun , Lee I-Ta , Lee Chiang-Wen , Chen Yuh-Lien 
Published: November 2023

AbstractBackgroundEpidemiology has demonstrated a strong relationship between fine particulate matter (PM) exposure and cardiovascular disease. Whether PM aggravates myocardial ischemia/reperfusion (I/R) injury and its related mechanisms remain unclear. Our previous study showed that adipose stem cell-derived exosomes (ADSC-Exo) contain a large amount of miR-221/222. This study investigated the effects of PM exposure on I/R-induced cardiac injury through mitophagy and apoptosis, as well as the potential role of miR-221/222 in ADSC-Exo.MethodsWild-type, miR-221/222 knockout (miR-221/222 KO), and miR-221/222 overexpressed transgenic (miR-221/222 TG) mice were intratracheally injected with 100 μg/kg PM for 24 h before I/R treatment. Ischemia was induced by temporarily occluding the left anterior descending (LAD) coronary artery with sutures for 30 min, followed by 3 h of reperfusion. In anin vitromodel, H9c2 cells were exposed to 50 μg/mL PM for 6 h and subjected to hypoxia (1% O2) at 37°C for 6 h, followed by 12 h reoxygenation.ResultsPM aggravates I/R (H/R)-induced cardiac injury by increasing ROS levels and causing mitochondrial dysfunction, leading to an increase in mitochondrial fission-related proteins like Drp1 and Mff, mitophagy-related proteins such as BNIP3 and LC3B, as well as apoptosis-related proteins like PUMA and p-p53in vivoandin vitrostudies. In comparison, transfection of ADSC-Exo and miR-221/222 mimics significantly reduced PM+I/R (H/R)-induced cardiac injury. Importantly, ADSC-Exo contains miR-221/222, which directly targets BNIP3, LC3B, and PUMA, decreasing their expression and ultimately reducing cardiomyocyte mitophagy and apoptosis.ConclusionsThe study showed that PM aggravates I/R or H/R-induced cardiac injury, and ADSC-Exo treatment significantly reduced this by regulating mitophagy and apoptosis through miR-221/222/BNIP3/LC3B/PUMA.
Clustered ProtocadherinCis-interactions are Required for Homophilic Combinatorial Cell-Cell Recognition Underlying Neuronal Self-Avoidance
Authors: Wiseglass Gil , Boni Nadir , Smorodinsky-Atias Karina , Rubinstein Rotem 
Published: November 2023

AbstractIn the developing human brain, only 53 stochastically expressed clustered protocadherin (cPcdh) isoforms enable neurites from an individual neuron to recognize and self-avoid, while maintaining contact with neurites from other neurons. Cell assays have demonstrated that self-recognition occurs only when all cPcdh isoforms perfectly match across the cell boundary, with a single mismatch in the cPcdh expression profile interfering with recognition. It remains unclear however, how a single mismatched isoform between neighboring cells, is sufficient to block erroneous recognitions. In using systematic cell aggregation experiments we show that abolishing cPcdh interactions on the same membrane (cis) results in a complete loss of specific combinatorial binding between cells (trans). Our computer simulations demonstrate that the organization of cPcdh in linear array oligomers, composed ofcisandtransinteractions, enhances self-recognition by increasing the concentration and stability of cPcdhtranscomplexes between the homotypic membranes. Importantly, we show that the presence of mismatched isoforms between cells drastically diminishes the concentrations and stability of thetranscomplexes. Overall, we provide an explanation for the role of the cPcdh assembly arrangements in neuronal self/non-self-discrimination underlying neuronal self-avoidance.
Project ChemicalBlooms: Collaborating with Citizen Scientists to Survey the Chemical Diversity and Phylogenetic Distribution of Plant Epicuticular Wax Blooms
Authors: Nguyen Dien , Groth Nicole , Mondloch Kylie , Cahoon Edgar B. , Jones Keith , Busta Lucas 
Published: November 2023

Plants use chemistry to overcome diverse challenges. A particularly striking chemical trait that some plants possess is the ability to synthesize massive amounts of epicuticular wax that accumulates on the plant’s surfaces as a white coating visible to the naked eye. The ability to synthesize basic wax molecules appears to be shared among virtually all land plants and our knowledge of ubiquitous wax compound synthesis is reasonably advanced. However, the ability to synthe-size thick layers of visible epicuticular crystals (“wax blooms”) is restricted to specific lineages and our knowledge of how wax blooms differ from ubiquitous wax layers is less developed. Here, we recruited the help of citizen scientists and middle school students to survey the wax bloom chemistry of 78 species spanning dicot, monocot, and gymnosperm lineages. Using gas chromatography-mass spectrometry, we found that the major wax classes reported from bulk wax mixtures can be present in wax bloom crystals, with fatty acids, fatty alcohols, and alkanes being present in many species’ bloom crystals. In contrast, other compounds including aldehydes, ketones, secondary alcohols, and triterpenoids were present in only a few species’ wax bloom crystals. By mapping the 78 wax bloom chemical profiles onto a phylogeny and using phylogenetic comparative analyses, we found that secondary alcohol and triterpenoid-rich wax blooms were present in lineage-specific patterns that would not be expected to arise by chance. That finding is consistent with reports that secondary alcohol biosynthesis enzymes are found only in certain lineages, but was a surprise for triterpenoids, which are intracellular components in virtually all plant lineages. Thus, our data suggest that a lineage-specific mechanism other than biosynthesis exists that enables select species to generate triterpenoid-rich surface wax crystals. Overall, our study outlines a general mode in which research scientists can collaborate with citizen scientists as well as middle and high school classrooms not only to enhance data collection and generate testable hypotheses, but also directly involve classrooms in the scientific process and inspire future STEM workers.Significance StatementPlants coat themselves in a protective layers of waxes. Some plants produce exceptionally thick layers of wax (“wax blooms”), and these thick layers are associated with enhanced abilities to tolerate stress, including drought and insect attack. In collaboration with citizen scientists and middle school classrooms, we provide an overview of the chemistry and phylogenetic distribution of plant epicuticular wax blooms. These data constitute a foundation upon which future studies of diverse wax blooms and their functions can build.
A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
Authors: Borao Sonia , Vega Montserrat , Boronat Susanna , Hidalgo Elena , Hümmer Stefan , Ayté José 
Published: November 2023

ABSTRACTSplicing is an important step of gene expression regulation in eukaryotes, as there are many mRNA precursors that can be alternatively spliced in different tissues, at different cell cycle phases or under different external stimuli. We have developed several life constructs that allow the quantification of fission yeast splicingin vivoon intact cells, and we have compared their splicing efficiency in a wild type strain and in aprp2-1(U2AF65) genetic background, showing a clear dependency between Prp2 and a consensus signal at 5’ splicing site (5’SS). To isolate novel genes involved in regulated splicing, we have crossed the reporter containing the weakest splicing rate with theSchizosaccharomyces pombeknock out collection. Among the candidate genes involved in the regulation of splicing, we have detected strong splicing defects in two of the mutants (Δcwf12andΔsaf5), both of them related to the NineTeen Complex (NTC). We have identified that strains with mutations incwf12have inefficient splicing, mainly when the 5’SS differs from the consensus. However, althoughΔsaf5cells also have some dependency on 5’SS sequence, we noticed that when one intron of a given pre-mRNA was affected, the rest of the introns of the same pre-mRNA had high probabilities of being also affected. This observation points Saf5 as a link between transcription rate and splicing.
Bounded solidarity? Experimental evidence on cross‐national bonding in the EU during the COVID crisis
Authors: OANA IOANA‐ELENA , TRUCHLEWSKI ZBIGNIEW 
Published: November 2023

AbstractMost studies on European solidarity (‘bonding’) during COVID‐19 lack a baseline comparison with outside states. We, therefore, cannot say whether European solidarity is universal or geared towards European Union (EU) insiders (‘bounding’). We thus ask whether European solidarity is ‘bounded’, that is, whether it relies on differentiation between European insiders and outsiders. We argue that if existent, bounded solidarity constitutes a long‐term and thick basis for institutional building. To explore this ‘bonding–bounding’ dynamic, we use a vignette experiment embedded into an original survey collected in eight European countries (n ∼ 8900), covering all European regions. Our design varies the countries receiving solidarity, and the channels (EU level vs. member state level), policy domains (health vs. economy) and instruments (loans, grants, medical equipment, vaccines) through which solidarity is provided. Regarding bounding, we find that most countries are more solidaristic with EU countries than an outsider, baseline state (Peru), Italy excepted. There is, nonetheless, a strong heterogeneity between countries: France, Germany, the Netherlands and Sweden mostly want to help themselves and Southern member states, but not Central Eastern European member states, which we show is due to their perceived obstructionism related to the Rule of Law debate. Concerning the nature of solidarity, we find that most respondents prefer solidarity to be channelled through the EU and loans, with evidence suggesting a freeriding mechanism behind this preference. All in all, our results indicate that EU citizens form a distinct community of solidarity which, in line with a Rokkanian understanding of polity formation, plays a key role in political development and consolidation.
Anaesthesia practice, quality indices including all‐cause 30‐day mortality associate to wrist fracture repositioning and surgery in Sweden: A perioperative register‐based study 2018–2021
Authors: Sellbrant Iren , Nellgård Bengt , Karlsson Jon , Albert Johanna , Jakobsson Jan G. 
Published: November 2023

AbstractBackgroundWrist fracture is one of most common fractures frequently requiring surgical anaesthesia. There is limited information related to the anaesthetic practice and quality including 30‐day mortality associated with wrist fracture in Sweden in recent years.AimThe aim of the present register‐based study was to investigate the anaesthesia techniques used and quality indices including 30‐day mortality associated with wrist fracture surgery in Sweden during the period 2018–2021.Materials and MethodsAll fracture repositions, and surgical interventions related to wrist fracture requiring anaesthesia in patients aged >18 years registered in the Swedish Perioperative Register (SPOR) between 2018 and 2021 were included in the analysis. Information on age, ASA class, anaesthesia technique, severe operative events, most reported side‐effects during recovery room stay and all‐cause 30‐day mortality was collected.ResultsThe data set included 25,147 procedures split into 14,796 females and 10,252 males (missing information n = 99) with a mean age of 52.9 ± 18.7 years and a significant age difference between females and males, 60.3 ± 15.4 and 42.2 ± 17.7 years, respectively. Mean age and ASA class increased during the study period (2018–2021), from 52.8 ± 18.6 to 54.0 ± 18.4 and ASA class 3–5 from 8.1% to 9.4% (p < .001 and p < .041, respectively). General anaesthesia (GA), GA combined with regional anaesthesia (RA), RA with or without sedation and sedation only was used in 41%, 13%, 40% and 6% of procedures, respectively, with minor changes over the study period. Pain at arrival in the recovery room (RR), (3.4%), severe pain during RR stay (2.1%), hypothermia (1.4%), postoperative nausea and vomiting (PONV) (1.2%) and urinary retention (0.5%) were the most reported side‐effects during the RR stay. (RA) was associated with significantly lower occurrence of pain and PONV, and shorter RR stay, compared with GA (p < .001). The all‐cause 30‐day mortality was low (19 of 25,147 (0.08%)) with no differences over the period studied or anaesthetic technique.ConclusionGeneral anaesthesia or general anaesthesia combined with regional anaesthesia are the most used anaesthetic techniques for wrist fracture procedures in Sweden. Recovery room pain, PONV, hypothermia and urinary retention is reported in overall low frequencies, with no change over the period studied, but in lower frequencies for regional anaesthesia. All‐cause 30‐day mortality was low; 0.08% with no change over time or between anaesthetic techniques. Thus, the present quality review based on SPOR data supports high quality of perioperative anaesthesia care.
Alcohol use disorder treatment via video conferencing compared with in‐person therapy during COVID‐19 social distancing : A non‐inferiority comparison of three cohorts
Authors: de Beurs Edwin , Rademacher Clara , Blankers Matthijs , Peen Jaap , Dekker Jack , Goudriaan Anneke 
Published: November 2023

AbstractBackgroundSocial distancing measures during the COVID‐19 pandemic forced an abrupt transformation of treatment delivery for mental health care. In mid‐March 2020, nearly all in‐person contact was replaced with video conferencing. The pandemic thus offered a natural experiment and a unique opportunity to conduct an observational study of whether alcohol use disorder treatment through video conferencing is non‐inferior to in‐person treatment.MethodsIn a large urban substance use disorder treatment center in the Netherlands, treatment evaluation is routine practice. Outcome data are regularly collected to support shared decision making and monitor patient progress. For this study, pre‐test and post‐test data on alcohol use (Measurements in the Addictions for Triage and Evaluation), psychopathology (Depression Anxiety Stress Scales), and quality of life (Manchester Short Assessment of Quality of Life) were used to compare outcomes of cognitive behavioral therapy treatment for three cohorts: patients who received treatment for a primary alcohol use disorder performed prior to (n = 628), partially during (n = 557), and entirely during (n = 653) the COVID‐19 lockdown.ResultsOutcome was similar across the three cohorts: No inferior outcomes were found for treatments that were conducted predominantly through video conferencing during lockdown or treatments that started in‐person, but were continued through video conferencing, compared to in‐person treatments that were conducted prior to COVID‐19. The number of drop‐outs were also similar between cohorts. However, there was a difference in average treatment intensity between cohorts, with treatment partially or fully conducted during the COVID‐19 pandemic lasting longer.ConclusionsTreatment for a primary alcohol use disorder, provided partially or predominantly through video conferencing during the COVID‐19 pandemic resulted in abstinence rates and secondary outcomes similar to traditional in‐person care, in spite of the potentially negative effects of the COVID‐related lockdown measures themselves. These results from everyday clinical practice corroborate findings of randomized controlled studies and meta‐analyses in which video conferencing appeared non‐inferior to in‐person care in clinical effectiveness.
Modeling the impact of a long‐term horizon and multiple treatment episodes on estimates of the cost‐effectiveness of alcohol treatment in the United States
Authors: Barbosa Carolina , Dowd William N. , Karriker‐Jaffe Katherine J. , Zarkin Gary 
Published: November 2023

AbstractBackgroundMost clinical studies of alcohol use disorder (AUD) treatment have short follow‐up periods, underestimating the full benefits of alcohol treatment. Furthermore, clinical studies only consider one treatment cycle and do not account for the need for multiple episodes to treat a chronic recurrent condition.MethodsA validated microsimulation model of the long‐term drinking patterns of people with AUD in the United States simulated 10,000 individuals resembling those from a large clinical trial. The model was used to assess the impact of (1) 1‐year, 5‐year, and lifetime horizon on alcohol treatment cost‐effectiveness estimates and (2) no, one, two, four, and unlimited additional treatment episodes on alcohol treatment cost‐effectiveness estimates. Model outcomes included healthcare costs, crime costs, labor market productivity, life expectancy, quality‐adjusted life years (QALYs), alcohol‐related hospitalizations, and deaths. Cost‐effectiveness analyses were conducted for two perspectives: a healthcare perspective that included costs from hospitalization and AUD treatment, and a broader societal perspective that also included crime costs and productivity.ResultsThe incremental cost per additional QALY gained for alcohol treatment compared with no treatment decreased from $55,590 after 1 year to $78 when healthcare costs and QALYs were tracked over the lifetime, that is, treatment became more cost effective. Treatment was cost saving for any time frame when the impacts on crime and labor productivity were also accounted for in a societal perspective. Access to multiple treatment episodes dominated (i.e., it was more effective and less costly) than no‐treatment and one‐episode scenarios. From a healthcare perspective, incremental costs per additional QALY for increasing from a maximum of two to four treatment episodes was $499 and from four to unlimited episodes was $5049. The unlimited treatment scenario dominated all others from a societal perspective. Results were robust in sensitivity analyses.ConclusionsA long‐term perspective and multiple episodes of alcohol treatment improve cost‐effectiveness estimates. When societal impacts are included, alcohol treatment is cost saving. Results support the value of alcohol treatment.
Retinopathy in Greyhound dogs: Prevalence, fundoscopic, and histopathological findings
Authors: Price Petra S. A. , Hunt Hayley , Cox Neil R. , Mitchell Nadia L. , Irving Arthur C. 
Published: November 2023

AbstractObjectivesTo determine the prevalence of retinal lesions and describe the fundoscopic findings of retinopathy in Greyhound dogs in the Manawatu/Whanganui region of New Zealand. To examine possible associations between sex, age, and racing variables with retinopathy in the study population. To describe retinal histologic findings in seven Greyhounds with retinopathy in New Zealand.MethodsTwo hundred Greyhound dogs from the Manawatu/Whanganui region of New Zealand underwent fundoscopy and fundic photography to identify and score the degree of retinopathy. Associations between retinopathy and age, sex, as well as racing variables, were examined. Histologic examination of the retina was undertaken on the eyes of seven Greyhounds from the Manawatu and Canterbury regions previously diagnosed with retinopathy by fundoscopy.ResultsFifty dogs (25.1%) were identified with retinopathy of varying degrees of severity. In at least one eye, 7.5% of dogs had mild retinopathy, 11.6% moderate retinopathy, and 6.0% severe retinopathy. Males were more likely to be affected in both eyes and with moderate or severe grades, than females. Increasing age was not associated with increased prevalence of retinopathy, nor increased grade of severity. Retinal histology identified multifocal retinal detachment in 5 of the 7 cases examined and other common lesions included choroidal necrosis and outer to full‐thickness retinal atrophy in the absence of significant inflammation.ConclusionsRetinopathy is prevalent in Greyhounds in the Manawatu/Whanganui region of New Zealand, but more research is required to elucidate the etiopathogenesis. Consideration should be made to include mandatory eye health examination in racing Greyhound dogs.
Efficacy of the positioning system when measuring canine intraocular pressures with the Reichert® Tono‐Vera® Vet rebound tonometer
Authors: Cabble Ava , Bajric Shayla , Kapeller Lydia E. , Harman Christine D. , Jacobson Amanda L. , Palanivel Hemalatha , Taylor David A. , Komáromy András M. 
Published: November 2023

AbstractObjectiveTo compare intraocular pressure (IOP) measurements in dogs taken with the Reichert® Tono‐Vera® Vet rebound tonometer with and without the automatic positioning system.Animals StudiedMeasurements were taken on 49 eyes from 26 Beagle‐derived dogs with variable genetics—four non‐glaucomatous and 22 ADAMTS10‐mutant dogs affected with different stages of open‐angle glaucoma. Seventeen of the 26 dogs were measured 2–4 times on different days with variable intervals since IOP‐lowering medications were administered.ProceduresIn each dog, tonometry was performed with the Tono‐Vera® Vet using three different methods in a randomized order: (Method 1) Average of three readings with an automatic positioning system; (Method 2) one reading with an automatic positioning system; and (Method 3) average of three readings obtained without the automatic positioning system. Statistical analyses included one‐way ANOVA, Tukey pairwise comparisons, and Bland–Altman plots (MiniTab®).ResultsWith each of the three tonometry methods, 116 measurements were taken, resulting in 348 total IOP measurements with a range of 12.8–49.9 mmHg. The means and standard deviations for each method were 25.4 ± 6.9 mmHg (Method 1), 26.0 ± 7.2 mmHg (Method 2), and 26.9 ± 7.7 mmHg (Method 3), with no significant differences (p = .27). Mean IOP variances were also not significantly different between tonometry methods (p = .24 to .78).ConclusionsBecause mean IOPs and their standard deviations were not statistically different between the three tonometry methods, we conclude that Tono‐Vera® Vet measurements conducted without the aid of the positioning system still provide reliable results.