bluuebunny/crossref_metadata_2025 huggingface dataset:
A Randomized, Double-Blind, Placebo Controlled Trial to Assess the Effectiveness and Safety of Melatonin and Three Formulations of Floraworks Proprietary TruCBN™ for Improving Sleep
Authors: Kolobaric Antonija , Saleska Jessica , Hewlings Susan J , Bryant Corey , Colwell Christopher S. , D’Adamo Christopher R. , Chen Jeff , Pauli Emily K
Published: November 2023

Introduction: Prescription sleep aids often have side effects, making alternative solutions necessary. Melatonin is an efficacious sleep aid and is often the comparator for alternatives. The phytocannabinoid cannabinol (CBN), has a potential mechanism of action but minimal evidence to support its effectiveness. Methods: TruCBN™ is a hemp-derived cannabinol (CBN) sleep aid. The primary aim of this randomized, double-blind, placebo-controlled study was to assess the safety and effects of three formulations of TruCBN™ [25 mg (n=206), 50 mg (n=205), 100 mg (n=203)], on sleep quality measured by PROMIS Sleep Disturbance 8A scale, relative to placebo (n=204). As a secondary aim, the effectiveness and safety of these formulations relative to a 4 mg melatonin (n=202) were assessed. Exploratory measures were stress (PROMIS Stress 4A), anxiety (Anxiety 4A), pain (PROMIS™ PEG) and well-being (WHO 5). Results: All treatment groups (25, 50 and 100 mg TruCBN™) and the 4 mg melatonin group experienced significant improvement in sleep quality relative to placebo. There were no significant differences between any of the TruCBN™ groups and melatonin for improving sleep. Participants taking 100 mg TruCBN™ showed a larger decrease in stress compared to the placebo group. There were no significant differences in anxiety, pain, well-being or frequency of side effects between any TruCBN™ group and placebo. There was no significant difference in improvements in sleep quality between any of the treatment groups and 4mg melatonin. Discussion: Orally ingested TruCBN™, 25 mg, 50 mg and 100 mg, is a safe and effective alternative for the improvement of sleep.

Sterling Institutional Review Board (SIRB) approved the study [10147-EKPauli]. The study was registered onClinicalTrials.gov[NCT05511818] on August 21, 2023.
Development and validation of artificial intelligence-based algorithms for predicting the segments debulked by rotational atherectomy using intravascular ultrasound
Authors: Hashimoto Kenta , Fujii Kenichi , Ueda Daiju , Sumiyoshi Akinori , Hasegawa Katsuyuki , Fukuhara Rei , Otagaki Munemitsu , Okamura Atsunori , Yamamoto Wataru , Kawano Naoki , Yamamoto Akira , Miki Yukio , Shiojima Iichiro
Published: November 2023

Background

Although rotation atherectomy (RA) is a useful technique for severely calcified lesions, patients undergoing RA show a greater incidence of catastrophic complications, such as coronary perforation. Therefore, prior to the RA procedure, it is important to predict which regions of the coronary plaque will be debulked by RA.

Objectives

We develop and evaluate an artificial intelligence–based algorithm that uses pre-RA intravascular ultrasound (IVUS) images to automatically predict regions debulked by RA

Methods

A total of 2106 IVUS cross-sections from 60 patients with de novo severely calcified coronary lesions who underwent IVUS-guided RA were consecutively collected. The two identical IVUS images of pre-and post-RA were merged, and the orientations of the debulked segments identified in the merged images are marked on the outer circle of each IVUS image. The artificial intelligence model was developed based on ResNet (deep residual learning for image recognition). The architecture connected 36 fully connected layers, each corresponding to one of the 36 orientations segmented every 10°, to a single feature extractor.

Results

In each cross-sectional analysis, our artificial intelligence model achieved an average sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 81%, 72%, 46%, 90%, and 75%, respectively.

Conclusions

The artificial intelligence–based algorithm can use information from pre-RA IVUS images to accurately predict regions debulked by RA. The proposed method will assist interventional cardiologists in determining the treatment strategies for severely calcified coronary lesions.
A CRISPR interference system for the nitrogen-fixing bacteriumAzotobacter vinelandii
Authors: Russell Steven J. , Garcia Amanda K. , Kaçar Betül
Published: November 2023

A grand challenge for the next century can be found in mitigating the effects of changing climate through bioengineering solutions. Biological nitrogen fixation, the globally consequential, nitrogenase-catalyzed reduction of atmospheric nitrogen to bioavailable ammonia, is a particularly vital area of focus. Nitrogen fixation engineering relies upon extensive understanding of underlying genetics in microbial models, including the broadly utilized gammaproteobacterium,Azotobacter vinelandii(A. vinelandii). Here we report the first CRISPR interference (CRISPRi) system for targeted gene silencing inA. vinelandiithat integrates genomically via site-specific transposon insertion. We demonstrate that CRISPRi can repress transcription of an essential nitrogen fixation gene by ∼60%. Further, we show that nitrogenase genes are suitably expressed from the transposon insertion site, indicating that CRISPRi and engineered nitrogen fixation genes can be co-integrated for combinatorial studies of gene expression and engineering. Our established CRISPRi system extends the utility ofA. vinelandiiand will aid efforts to engineer microbial nitrogen fixation for desired purposes.

For Table of Contents Only
The five homologous CiaR-controlled Ccn sRNAs ofStreptococcus pneumoniaemodulate Zn-resistance
Authors: De Lay Nicholas R. , Verma Nidhi , Sinha Dhriti , Garrett Abigail , Osterberg Maximillian K. , Reiling Spencer , Porter Daisy , Giedroc David P. , Winkler Malcolm E.
Published: November 2023

Zinc is a vital transition metal forStreptococcus pneumoniae, but is deadly at high concentrations. InS. pneumoniae, elevated intracellular free Zn levels result in mis-metallation of key Mn-dependent metabolic and superoxide detoxifying enzymes resulting in Zn intoxication. Here, we report our identification and characterization of the function of the five homologous, CiaRH-regulated Ccn sRNAs in controllingS. pneumoniaevirulence and metal homeostasis. We show that deletion of all fiveccngenes (ccnA,ccnB,ccnC,ccnD, andccnE) fromS. pneumoniaestrains D39 (serotype 2) and TIGR4 (serotype 4) causes Zn hypersensitivity and an attenuation of virulence in a murine invasive pneumonia model. We provide evidence that bioavailable Zn disproportionately increases inS. pneumoniaestrains lacking the fiveccngenes. Consistent with a response to Zn intoxication or relatively high intracellular free Zn levels, expression of genes encoding the CzcD Zn exporter and the Mn-independent ribonucleotide reductase, NrdD-NrdG, were increased in the ΔccnABCDEmutant relative to its isogenicccn+parent strain. The growth inhibition by Zn that occurs as the result of loss of theccngenes is rescued by supplementation with Mn or OxyraseTM, a reagent that removes dissolved oxygen. Lastly, we found that the Zn-dependent growth inhibition of the ΔccnABCDEstrain was not altered by deletion ofsodA, whereas theccn+ΔsodAstrain phenocopied the ΔccnABCDEstrain. Overall, our results indicate that the Ccn sRNAs have a crucial role in preventing Zn intoxication inS. pneumoniae.

AUTHOR SUMMARY

Zn and Mn are essential micronutrients for many bacteria, includingStreptococcus pneumoniae. While Zn performs vital structural or catalytic roles in certain proteins, in excess, Zn can inhibit Mn uptake byS. pneumoniaeand displace, but not functionally replace Mn from key enzymes including superoxide dismutase A (SodA). Here, we show that the Ccn small regulatory RNAs promoteS. pneumoniaeresistance to Zn intoxication. Furthermore, we demonstrate that these small regulatory RNAs modulate the ability ofS. pneumoniaeto cause invasive pneumonia. Altogether, these findings reveal a new layer of regulation ofS. pneumoniaeZn homeostasis and suggest that there are factors in addition to known transporters that modulate intracellular, bioavailable Zn levels.
Craniofacial studies in chicken embryos confirm the pathogenicity of Frizzled2 variants associated with Robinow syndrome
Authors: Tophkhane Shruti S. , Fu Katherine , Verheyen Esther M. , Richman Joy M.
Published: November 2023

Robinow syndrome (RS) is a rare disease caused by mutations in seven WNT pathway genes. Features include craniofacial widening and jaw hypoplasia. We used the chicken embryo to test two autosomal dominant RS (ADRS) missenseFZD2variants on the frontonasal mass, the affected region in RS. The wild-type (wt) and variant hFZD2inhibited beak ossification. The bone hypoplasia was possibly mediated by decreased levels of WNT and BMP pathway genes. In primary cultures, hFZD2variants inhibited chondrogenesis, increased nuclear shuttling of β-catenin and increased expression of TWIST1, both known to suppress chondrogenesis. In luciferase reporter assays, proteins coding for1301G>Tand425C>TFZD2 variants weakly activated canonical WNT reporter and dominantly interfered with wtFZD2. In the JNK-PCP WNT pathway luciferase assay, only the425C>Tshowed a loss-of-function. The 1301G>T variant presumably acts through a JNK-independent pathway. This is the first study to demonstrate that the ADRS-FZD2missense variants cause craniofacial and WNT signaling defects. Frontonasal mass width is increased by both hFZD2variants which sheds light on the ontogeny of the broad facial features seen in individuals with RS.

Summary Statement

Gain-of-function studies onFZD2missense variants associated with Robinow syndrome led to increased facial width, altered Wnt signaling and inhibition of beak skeletogenesis in chicken embryos.
Integrative learning of disentangled representations from single-cell RNA-sequencing datasets
Authors: Novella-Rausell Claudio , Peters D.J.M , Mahfouz Ahmed
Published: November 2023

Abstract

Single-cell RNA-sequencing is instrumental in studying cellular diversity in biological systems. Using batch correction methods, cell identities are often jointly defined across multiple conditions, individuals, or modalities. These approaches overlook group-specific information and require either paired data or matching features across datasets. Here we present shared-private Variational Inference via Product of Experts with Supervision (spVIPES), a framework to analyze the shared and private components of unpaired groups of cells with non-matching features. spVIPES represents the cells from the different groups as a composite of private and shared factors of variation using a probabilistic latent variable model. We evaluate the performance of spVIPES with a simulated dataset and apply our model in three different scenarios: (i) cross-species comparisons, (ii) regeneration following long and short acute kidney injury, and (iii) IFN-βstimulation of PMBCs. In our study, we demonstrate that spVIPES accurately disentangles distinct sources of variation into private and shared representations while matching current state-of-the-art methods for batch correction. Furthermore, spVIPES’ shared space outperforms alternatives models at learning cell identities across datasets with non-matching features. We implemented spVIPES using thescvi-toolsframework and release it as an open-source software athttps://github.com/nrclaudio/spVIPES.
Bacteriophages from human skin infecting coagulase-negativeStaphylococcus:diversity, novel species and host resistance
Authors: Alsaadi Samah E. , Lu Hanshuo , Zhang Minxing , Dykes Gregory F. , Allison Heather E. , Horsburgh Malcolm J.
Published: November 2023

The human skin microbiome comprises diverse populations that differ temporally between body sites and individuals. The virome is a less studied component of the skin microbiome and the study of bacteriophages is required to increase knowledge of the modulation and stability of bacterial communities.Staphylococcusspecies are among the most abundant colonisers of skin and are associated with both health and disease yet the bacteriophages infecting the most abundant species on skin are less well studied. Here, we report the isolation and genome sequencing of 40 bacteriophages from human skin swabs that infect coagulase-negativeStaphylococcus(CoNS) species, which extends our knowledge of phage diversity. Six genetic clusters of phages were identified with two clusters representing novel phage species, one of which we characterise and name Alsa phage. We identified that Alsa phages have a greater ability to infect the speciesS. hoministhat was otherwise infected less than other CoNS species by the isolated phages, indicating an undescribed barrier to phage infection that could be in part due to numerous restriction-modification systems. The extended diversity ofStaphylococcusphages here enables further research to define their contribution to skin microbiome research and the mechanisms that limit phage infection.
Astrocyte-to-microglia communication via Sema4B-Plexin-B2 modulates injury-induced reactivity of microglia
Authors: Casden Natania , Belzer Vitali , Elkhayari Abdellatif , Elfatimy Rachid , Behar Oded
Published: November 2023

After central nervous system injury, a rapid cellular and molecular response is induced. This response can be both beneficial and detrimental to neuronal survival in the first few days and increases the risk for neurodegeneration if persistent. Semaphorin4B (Sema4B), a transmembrane protein primarily expressed by cortical astrocytes, has been shown to play a role in neuronal cell death following injury. Our study shows that after cortical stab wound injury, cytokine expression is attenuated in Sema4B knockout mice and microglia/macrophage reactivity is altered.In vitro, Sema4B enhances the reactivity of microglia following injury, suggesting astrocytic Sema4B functions as a ligand. Moreover, injury-induced microglia reactivity is attenuated in the presence of Sema4B knockout astrocytes compared to heterozygous astrocytes.In vitro, experiments indicate Plexin-B2 is the Sema4B receptor on microglia. Consistent with this, in microglia/macrophage-specific Plexin-B2 knockout mice, similar to Sema4B knockout mice, microglial/macrophage reactivity and neuronal cell death are attenuated after cortical injury. Finally, in Sema4B/Plexin-B2 double heterozygous mice, microglial/macrophage reactivity is also reduced after injury, thus supporting the idea that both Sema4B and Plexin-B2 are part of the same signaling pathway. Taken together, we propose a model in which following injury, astrocytic Sema4B enhances the response of microglia/macrophages via Plexin-B2, leading to increased reactivity.

Significance statement

In this study, we show that in the brain cortex, Sema4B, a protein mainly expressed by astrocytes, plays a crucial role in enhancing the reactivity of microglia/macrophages via Plexin-B2. These findings reveal new molecular signaling instigated by astrocytes toward microglia/macrophages in the context of central nervous system (CNS) injury, shedding new light on the complex interplay between astrocytes and microglia/macrophages. Taken together, our findings suggest that targeting the Sema4B/Plexin-B2 pathway could be a promising therapeutic approach for reducing microglia reactivity and improving the adaptive response in the context of CNS injury.
Predicting human and viral protein variants affecting COVID-19 susceptibility and repurposing therapeutics
Authors: Waman Vaishali P. , Ashford Paul , Lam Su Datt , Sen Neeladri , Abbasian Mahnaz , Woodridge Laurel , Goldtzvik Yonathan , Bordin Nicola , Wu Jiaxin , Sillitoe Ian , Orengo Christine A
Published: November 2023

The COVID-19 disease is an ongoing global health concern. Although vaccination provides some protection, people are still susceptible to re-infection. Ostensibly, certain populations or clinical groups may be more vulnerable. Factors causing these differences are unclear and whilst socioeconomic and cultural differences are likely to be important, human genetic factors could influence susceptibility. Experimental studies indicate SARS-CoV-2 uses innate immune suppression as a strategy to speed-up entry and replication into the host cell. Therefore, it is necessary to understand the impact of variants in immunity-associated human proteins on susceptibility to COVID-19.

In this work, we analysed missense coding variants in several SARS-CoV-2 proteins and its human protein interactors that could enhance binding affinity to SARS-CoV-2. We curated a dataset of 19 SARS-CoV-2: human protein 3D-complexes, from the experimentally determined structures in the Protein Data Bank and models built using AlphaFold2-multimer, and analysed impact of missense variants occurring in the protein-protein interface region. We analysed 468 missense variants from human proteins and 212 variants from SARS-CoV-2 proteins and computationally predicted their impacts on binding affinities to SARS-CoV-2 proteins, using 3D-complexes.

We predicted a total of 26 affinity-enhancing variants from 14 human proteins implicated in increased binding affinity to SARS-CoV-2. These include key-immunity associated genes (TOMM70, ISG15, IFIH1, IFIT2, RPS3, PALS1, NUP98, RAE1, AXL, ARF6, TRIMM, TRIM25) as well as important spike receptors (KREMEN1, AXL and ACE2). We report both common (e.g., Y13N in IFIH1) and rare variants in these proteins and discuss their likely structural and functional impact, using information on known and predicted functional sites. Potential mechanisms associated with immune suppression implicated by these variants are discussed.

Occurrence of certain predicted affinity-enhancing variants should be monitored as they could lead to increased susceptibility and reduced immune response to SARS-CoV-2 infection in individuals/populations carrying them. Our analyses aid in understanding the potential impact of genetic variation in immunity-associated proteins on COVID-19 susceptibility and help guide drug-repurposing strategies.
Unravelling the Role ofCandida albicansPrn1 in the Oxidative Stress Response Through a Proteomic Approach
Authors: Arribas Víctor , Monteoliva Lucía , Hernáez María Luisa , Gil Concha , Molero Gloria
Published: November 2023

Candida albicansPrn1 is a protein that shares similarities with mammalian Pirin but with an unknown function in the yeast. Orthologues of Prn1 have been identified in other pathogenic fungi but not inSaccharomyces cerevisiae, suggesting a relationship with pathogenesis. Prn1 increase in abundance after H2O2treatment has been shown previously, thus, in the present work,C. albicans prn1Δmutant and the corresponding wild-type strain SN250 have been treated with H2O2and their response was studied by quantitative differential proteomics. These assays indicated a lower increase of proteins with oxidoreductase activity after treatment in theprn1Δstrain compared to the wild type, as well as an increase in proteasome-activating proteins and a decrease in translation-involved proteins. Accordingly, Prn1 absence, under H2O2treatment, led to a lower survival rate and a higher percentage of apoptosis, together with higher reactive oxygen species levels and higher proteasome activity. Besides, remarkable differences in the abundance of some transcription factors were observed between the two strains. Mnl1, involved in Prn1 expression, Bas1, Tiff33, and orf19.1150 presented an inverse pattern of expression under H2O2treatment respect to Nrg1, a Mnl1 antagonist. Interestingly, orf19.4850, a protein orthologue toS. cerevisiaeCub1, has shown to be involved in the response to H2O2presenting a conserved proteasome function. Under basal conditions, the proteomics results indicate a possible involvement of Prn1 in mitochondrial oxidative stress detoxication. Our experiments confirm Prn1 as a relevant actor in the oxidative response.

Importance

Candida albicansis a human opportunistic pathogen included in the WHO fungal priority pathogens list. The increase in resistant strains necessitates the discovery of new targets for antifungal therapies. Our research sheds light on the important role of the previously uncharacterizedC. albicansprotein Prn1 during the oxidative stress response. Study of the proteome remodelling under oxidative stress unveils the role of Prn1 in the decreased reactive oxygen species levels and the consequences, such as death by apoptosis and necrosis or cell growth delay. A proteomics approach allowed the identification of several proteins potentially involved in Prn1 activity, such as oxidoreductases and transcription factors. The lack of Prn1 orthologues inSaccharomyces cerevisiaebut the presence in otherCandidaandAspergillusspecies implicates this protein in pathogenesis and suggests that it may serve as a candidate for new drug targets.
Single-cell transcriptomic and genomic changes in the aging human brain
Authors: Jeffries Ailsa M. , Yu Tianxiong , Ziegenfuss Jennifer S. , Tolles Allie K. , Kim Yerin , Weng Zhiping , Lodato Michael A.
Published: November 2023

Aging brings dysregulation of various processes across organs and tissues, often stemming from stochastic damage to individual cells over time. Here, we used a combination of single-nucleus RNA-sequencing and single-cell whole-genome sequencing to identify transcriptomic and genomic changes in the prefrontal cortex of the human brain across life span, from infancy to centenarian. We identified infant-specific cell clusters enriched for the expression of neurodevelopmental genes, and a common down-regulation of cell-essential homeostatic genes that function in ribosomes, transport, and metabolism during aging across cell types. Conversely, expression of neuron-specific genes generally remains stable throughout life. We observed a decrease in specific DNA repair genes in aging, including genes implicated in generating brain somatic mutations as indicated by mutation signature analysis. Furthermore, we detected gene-length-specific somatic mutation rates that shape the transcriptomic landscape of the aged human brain. These findings elucidate critical aspects of human brain aging, shedding light on transcriptomic and genomics dynamics.
Kaposi’s sarcoma-associated herpesvirus (KSHV) gB dictates a low-pH endocytotic entry pathway as revealed by a dual-fluorescent virus system and a rhesus monkey rhadinovirus expressing KSHV gB
Authors: Schlagowski Sarah , Großkopf Anna K. , Liu Shanchuan , Khizanishvili Natalia , Scribano Stefano , Ensser Armin , Hahn Alexander S.
Published: November 2023

Interaction with host cell receptors initiates internalization of Kaposi’s sarcoma-associated herpesvirus (KSHV) particles via endocytosis or macropinocytosis. Fusion of viral and host cell membranes, which is followed by release of the viral capsid into the cytoplasm, is executed by the core fusion machinery composed of gH, gL, and gB, that is common to all herpesviruses. KSHV infection has been shown to be sensitive to inhibitors of vacuolar acidification, suggestive of low pH as a fusion trigger.

To analyze KSHV entry at the single particle level we developed single- and dual-fluorescent recombinant KSHV strains that incorporate tagged glycoproteins, capsid proteins or a combination thereof. In addition, we generated a hybrid rhesus monkey rhadinovirus (RRV) that expresses KSHV gB in place of RRV gB to analyze gB-dependent differences in infection pathways.

Our data demonstrate lytic reactivation and infectivity of dual-fluorescent KSHV and incorporation of fluorescently-tagged proteins into viral particles. Confocal microscopy was used to quantify co-localization of fluorescently-tagged glycoproteins and capsid proteins. By using the ratio of dual-positive KSHV particles to single-positive capsids as an indicator of fusion events we established the KSHV fusion kinetics upon infection of different target cells.

We measured marked differences in the “time-to-fusion” between different cell types. Inhibition of vesicle acidification prevented virus-cell fusion, implicating low vesicle pH as a requirement for the KSHV fusion step. These findings were corroborated by comparison of RRV-YFP reporter virus with wildtype gB and RRV-YFP encoding KSHV gB in place of RRV gB. While RRV wt infection of receptor-overexpressing cells was unaffected by inhibition of vesicle acidification, RRV-YFP expressing KSHV gB was sensitive to Bafilomycin A1, an inhibitor of vacuolar acidification.

Single- and dual-fluorescent KSHV strains eliminate the need for virus-specific antibodies and enable the tracking of single viral particles during entry and fusion. Together with a hybrid RRV expressing KSHV gB, these two novel tools identify low vesicle pH as an endocytotic trigger for KSHV membrane fusion.
New tools reveal PCP-dependent polarized mechanics in the cortex and cytoplasm of single cells during convergent extension
Authors: Weng Shinuo , Devitt Caitlin C. , Nyaoga Bill M. , Havnen Anna E. , Alvarado José , Wallingford John B.
Published: November 2023

Understanding biomechanics of biological systems is crucial for unraveling complex processes like tissue morphogenesis. However, current methods for studying cellular mechanicsin vivoare limited by the need for specialized equipment and often provide limited spatiotemporal resolution. Here we introduce two new techniques, Tension by Transverse Fluctuation (TFlux) andin vivomicrorheology, that overcome these limitations. They both offer time-resolved, subcellular biomechanical analysis using only fluorescent reporters and widely available microscopes. Employing these two techniques, we have revealed a planar cell polarity (PCP)-dependent mechanical gradient both in the cell cortex and the cytoplasm of individual cells engaged in convergent extension. Importantly, the non-invasive nature of these methods holds great promise for its application for uncovering subcellular mechanical variations across a wide array of biological contexts.

Non-invasive imaging-based techniques providing time-resolved biomechanical analysis at subcellular scales in developing vertebrate embryos.
VPS8D, a CORVET subunit, is required to maintain the contractile vacuole complex inTetrahymena thermophila
Authors: Cheng Chao-Yin , Hernández Josefina , Turkewitz Aaron P.
Published: November 2023

Contractile vacuole complexes (CVCs) are complex osmoregulatory organelles, with vesicular (bladder) and tubular (spongiome) subcompartments. The mechanisms that underlie their formation and maintenance within the eukaryotic endomembrane network are poorly understood. In the CiliateTetrahymena thermophila, six differentiated CORVETs (class C core vacuole/endosome tethering complexes), with Vps8 subunits designated A-F, are likely to direct endosomal trafficking. Vps8Dp localizes to both bladder and spongiome. We show by inducible knockdown thatVPS8Dis essential to CVC organization and function.VPS8Dknockdown increased susceptibility to osmotic shock, tolerated in the wildtype but triggering irreversible lethal swelling in the mutant. The knockdown rapidly triggered contraction of the spongiome and lengthened the period of the bladder contractile cycle. More prolonged knockdown resulted in disassembly of both the spongiome and bladder, and dispersal of proteins associated with those compartments. In stressed cells where the normally singular bladder is replaced by numerous vesicles bearing bladder markers, Vps8Dp concentrated conspicuously at long-lived inter-vesicle contact sites, consistent with tethering activity. Similarly, Vps8Dp in cell-free preparations accumulated at junctions formed after vacuoles came into close contact. Also consistent with roles for Vps8Dp in tethering and/or fusion were the emergence in knockdown cells of multiple vacuole-related structures, replacing the single bladder.

Synopsis

In the Ciliate Tetrahymena thermophila,VPS8D, which encodes a subunit of a non-conventional CORVET complex, is an essential determinant of the contractile vacuole complex (CVC). VPS8D knockdown results in retraction and dispersal of the spongiome, and disappearance of the bladder, reinforcing the view that CVCs arise from endosomal trafficking. Intermediate knockdown phenotypes and Vps8Dp localization support a role in homotypic tethering.
An interaction hub on Ndc80 complex facilitates dynamic recruitment of Mps1 to yeast kinetochores to promote accurate chromosome segregation
Authors: Parnell Emily J. , Jenson Erin , Miller Matthew P.
Published: November 2023

Accurate chromosome segregation relies on kinetochores carrying out multiple functions, including establishing and maintaining microtubule attachments, forming precise bioriented attachments between sister chromatids, and activating the spindle assembly checkpoint. Central to these processes is the highly conserved Ndc80 complex. This kinetochore subcomplex interacts directly with microtubules, but also serves as a critical platform for recruiting kinetochore-associated factors and as a key substrate for error correction kinases. The precise manner in which these kinetochore factors interact, and regulate each other’s function, remains unknown – considerably hindering our understanding of how Ndc80 complex-dependent processes function together to orchestrate accurate chromosome segregation. Here, we aimed to uncover the role of Nuf2’s CH domain, a component of the Ndc80 complex, in ensuring accurate chromosome segregation. Through extensive mutational analysis, we identified a conserved “interaction hub” comprising two segments in Nuf2’s CH domain, forming the binding site for Mps1 within the yeast Ndc80 complex. Intriguingly, the interaction between Mps1 and the Ndc80 complex seems to be subject to regulation by competitive binding with other factors. Mutants disrupting this interaction hub exhibit defects in spindle assembly checkpoint function and severe chromosome segregation errors. Significantly, specifically restoring Mps1-Ndc80 complex association rescues these defects. Our findings shed light on the intricate regulation of Ndc80 complex-dependent functions and highlight the essential role of Mps1 in kinetochore biorientation and accurate chromosome segregation.
Thermal imaging can reveal variation in stay-green functionality of wheat canopies under temperate conditions
Authors: Anderegg Jonas , Kirchgessner Norbert , Aasen Helge , Zumsteg Olivia , Keller Beat , Zenkl Radek , Walter Achim , Hund Andreas
Published: November 2023

Canopy temperature (CT) is often interpreted as representing leaf activity traits such as photosynthetic rates, gas exchange rates, or stomatal conductance. Accordingly, CT measurements may provide a basis for high throughput assessments of the productivity of wheat canopies during early grain filling, which would allow distinguishing functional from dysfunctional stay-green. However, whereas the usefulness of CT as a fast surrogate measure of sustained vigor under soil drying is well established, its potential to quantify leaf activity traits under high-yielding conditions is less clear. To better understand sensitivity limits of CT measurements under high yielding conditions, we generated within-genotype variability in stay-green functionality by means of differential short-term pre-anthesis canopy shading that modified the sink:source balance. We quantified the effects of these modifications on stay-green properties through a combination of gold standard physiological measurements of leaf activity and newly developed methods for organ-level senescence monitoring based on timeseries of high-resolution imagery and deep-learning-based semantic image segmentation. In parallel, we monitored CT by means of a pole-mounted thermal camera that delivered continuous, ultra-high temporal resolution CT data. Our results show that differences in leaf activity stemming from differences in stay-green functionality translate into measurable differences in CT in the absence of major confounding factors. Differences amounted to approximately 0.8°C and 1.5°C for a very high-yielding source-limited genotype, and a medium-yielding sink-limited genotype, respectively. The gradual nature of the effects of shading on CT during the stay-green phase underscore the importance of a high measurement frequency and a time-integrated analysis of CT, whilst modest effect sizes confirm the importance of restricting screenings to a limited range of morphological and phenological diversity.
Voice and “digital disruption” in internal communication job advertisements: proposing a vocality continuum
Authors: Sandham Sonya
Published: November 2023

Purpose

This study of job advertisements for internal communication practitioners aims to investigate the signals that organisations are sending the profession about what is required of these roles. The concept of corporate voice – the “voice” of the organisation – is problematised to explore tensions in vocality. The aim is to support communication practitioners to navigate multi-vocality in the evolving professional context of digital communication technologies and changes in the workplace.

Design/methodology/approach

This qualitative study considers the role of voice in corporate communication practices and offers insights into “digital disruption” and the discursive pressure of employers' priorities on the profession and its practices. Job advertisements for internal communication practitioners were examined during 6-month periods in 2018, 2020 and 2022, which was a significant time of change for the profession with the global COVID-19 pandemic.

Findings

Qualitative content analysis of 514 internal communication job advertisements identifies that control and consistency are valorised, and continue to dominate descriptions of internal communication skills and responsibilities. The digital affordances that communication practitioners rely on has not changed significantly and a preference for “broadcasting” is evident.

Originality/value

This study provides insights into how Australian organisations shape and sustain univocal corporate communication practices, and the incompatibility of narrow configurations of voice with emerging organisational challenges such as social connectedness.
The mediating effect of patient trust on the relationship between service quality and patient satisfaction
Authors: Al-hilou Mais , Suifan Taghrid
Published: November 2023

Purpose

The prime aim of this research is to examine the mediating effect of patient trust on the relationship between service quality and patient satisfaction in Amman, Jordan.

Design/methodology/approach

A convenience sample of patients visiting seven out of the 44 private hospitals in Amman was selected. In total, 385 questionnaires were distributed among patients, with a response rate of 91%. Of these, 35 were disregarded, and the data from the remaining 350 questionnaires were analyzed using SPSS.

Findings

The results showed that service quality has a statistically significant effect on patient satisfaction. Furthermore, service quality has a statistically significant effect on patient trust, while there is also a statistically significant effect of patient trust on patient satisfaction. The findings also revealed that patient satisfaction partially mediates the relationship between service quality and patient satisfaction.

Originality/value

Generating more insights in the areas of service quality, patient trust and patient satisfaction while also extending the findings of earlier studies. The prior studies in the literature that focus on customers are given a more advantageous perspective by using Jordanian hospitals as a population to test the model of this research. The majority of past research on service quality, patient satisfaction and patient trust in the healthcare sector has been conducted in western nations.
The effect of pretranslation techniques in developing EFL students' translation ability
Authors: Bin-Hady Wagdi Rashad Ali , Al-Ahdal Arif Ahmed Mohammed Hassan , Abdullah Samia Khalifa
Published: November 2023

Purpose

English as a foreign langauge (EFL) students find it difficult to apply the theoretical knowledge they acquire on translation in the practical world. Therefore, this study explored if training in pretranslation techniques (PTTs) (syntactic parsing) as suggested by Almanna (2018) could improve the translation proficiency of Yemeni EFL students. Moreover, the study also assessed which of the PTTs the intervention helped to develop.

Design/methodology/approach

The study adopted a primarily experimental pre- and posttests research design, and the sample comprised of an intake class with 16 students enrolled in the fourth year, Bachelor in Education (B.Ed), Hadhramout University. Six participants were also interviewed to gather the students' perceptions on using PTTs.

Findings

Results showed that students' performance in translation developed significantly (Sig. = 0.002). All the six PTTs showed development, though subject, tense and aspect developed more significantly (Sig. = 0.034, 0.002, 0.001 respectively). Finally, the study reported students' positive perceptions on the importance of using PTTs before doing any translation tasks.

Originality/value

One of the recurrent errors that can be noticed in Yemeni EFL students' production is their inability to transfer the grammatical elements of sentences from L1 (Arabic) into L2 (English) or the visa versa. The researchers thought though translation is more than the syntactic transmission of one language into another, analyzing the elements of sentences using syntactic and semantic parsing can help students to produce acceptable texts in the target language. These claims would be proved or refuted after analyzing the experiment result of the present study.
How can we help law enforcement agencies learn? A look at CALEA police accreditation
Authors: Abner Gordon , Merritt Cullen C. , Boggs Rachel
Published: November 2023

Purpose

This study explores the benefits of accreditation from the Commission on Accreditation for Law Enforcement Agencies (CALEA), according to those who are engaged in the practice.

Design/methodology/approach

The authors collected data through open-ended, semi-structured telephone interviews. The authors analyzed the data using an inductive methodology.

Findings

The authors found that CALEA police accreditation enhances organizational learning through the development of knowledge brokers, the creation of communities of practice, support for knowledge repositories, support for knowledge managers and greater levels of transparency.

Originality/value

This qualitative study, which focuses on the perceptions and experiences of those involved in the CALEA process, provides a valuable complement to the quantitative literature on accreditation by shedding light on the organizational learning resulting from accreditation.
Is it time to rethink benefit packages? Perks associated with the intention to leave in different age groups during COVID-19
Authors: Masso Piret , Jaakson Krista , Põder Kaire
Published: November 2023

Purpose

The study's objective is to estimate the association of specific perceived employer-provided benefits on employees' intention to leave in different age cohorts during coronavirus disease 2019 (COVID-19). Informed by the psychological theories of ageing, the authors propose three age-cohort-specific hypotheses in three motivational domains: security and health benefits, flexible work arrangement and education-related benefits.

Design/methodology/approach

The authors use a large survey of employees in Estonia (n = 7,209) conducted in 2020 and test the association of specific benefits and their interactions with age on employees' intention to leave.

Findings

The results show that older cohorts are generally less prone to leave their jobs. Benefits that employers could use during the COVID-19 crisis generally had negative associations with the intention to leave, but age-specific differences were negligible; only the perceived provision of flexible work arrangements reduced the younger cohort's intention to leave relatively more.

Originality/value

This study is one of the few that allows us to make inferences regarding the benefits preferences amongst the working population during an unprecedented health crisis.
SIRT6 deficiency causes ovarian hypoplasia by affecting Plod1‐related collagen formation
Authors: Li Liyuan , Hua Rui , Hu Kaiqiang , Chen Huiling , Yin Yuemiao , Shi Xiaojin , Peng Kezheng , Huang Qing , Qiu Ying , Li Xue , Liu Qingfei , Liu Shangfeng , Wang Zhao
Published: November 2023

SIRT6 is a key member of the mammalian sirtuin family of conserved nicotinamide adenine dinucleotide (NAD+)‐dependent deacetylases. Previous studies have shown that SIRT6 can regulate metabolism, DNA damage repair and aging. Ovarian aging process usually share similar mechanisms with general aging, which is characterized by decreases in both numbers of ovarian follicles and the quality of oocytes. It is reported that the expression level of SIRT6 was significantly decreased in the ovaries of aged mice, and the level of SIRT6 was positively correlated with ovarian reserve, indicating that SIRT6 may be potential markers of ovarian aging. However, its biological roles in follicular development are still unclear. Here, we explored the effect of SIRT6 on follicular development and found that ovarian development was interrupted in SIRT6 knockout (KO) mice, leading to disruptions of puberty and the estrus cycle, significant decreases in numbers of secondary and antral follicles, and decreased collagen in the ovarian stroma. Plod1, a lysyl hydroxylase that is vital for collagen crosslinking and deposition, was decreased at both the mRNA and protein levels in SIRT6‐deficient ovaries and granulosa cells (GCs). Additionally, we found abnormal estrogen levels in both SIRT6 KO mice and SIRT6 KD GCs, accompanied by decreases in the levels of the estrogen biosynthesis genes Cyp11a1, Cyp19a1, Mgarp, and increases in the levels of TNF‐α and NF‐κB. These results confirmed the effect of SIRT6 on follicular development and revealed a possible molecular mechanism for SIRT6 involvement in follicular development via effects on estrogen biosynthesis and collagen formation.
Rapid appearance of negative emotion during oral fentanyl self‐administration in male and female rats
Authors: Coffey Kevin R. , Nickelson William B. , Dawkins Aliyah J. , Neumaier John F.
Published: November 2023

Opioid use disorder has become an epidemic in the United States, fuelled by the widespread availability of fentanyl, which produces rapid and intense euphoria followed by severe withdrawal and emotional distress. We developed a new preclinical model of fentanyl seeking in outbred male and female rats using volitional oral self‐administration (SA) that can be readily applied in labs without intravascular access. Using a traditional two‐lever operant procedure, rats learned to take oral fentanyl vigorously, escalated intake across sessions, and readily reinstated responding to conditioned cues after extinction. Oral SA also revealed individual and sex differences that are essential to studying substance use risk propensity. During a behavioural economics task, rats displayed inelastic demand curves and maintained stable intake across a wide range of fentanyl concentrations. Oral SA was also neatly patterned, with distinct ‘loading’ and ‘maintenance’ phases of responding within each session. Using our software DeepSqueak, we analysed ultrasonic vocalizations (USVs), which are innate expressions of current emotional state in rats. Rats produced 50 kHz USVs during loading then shifted quickly to 22 kHz calls despite ongoing maintenance of oral fentanyl taking, reflecting a transition to negative reinforcement. Using fibre photometry, we found that the lateral habenula differentially processed drug cues and drug consumption depending on affective state, with potentiated modulation by drug cues and consumption during the negative affective maintenance phase. Together, these results indicate a rapid progression from positive to negative reinforcement occurs even within an active drug taking session, revealing a within‐session opponent process.
Open‐label investigation of rapid initiation of extended‐release buprenorphine in patients using fentanyl and fentanyl analogs
Authors: Mariani John J. , Dobbins Robert L. , Heath Amy , Gray Frank , Hassman Howard
Published: November 2023

Background and Objectives

Synthetic opioids, including fentanyl and fentanyl analogs, account for over 70,000 annual overdose deaths in the United States, but there is limited information examining methods of induction and maintenance outcomes for buprenorphine treatment of patients with opioid use disorder (OUD) using these opioids.

Methods

A secondary analysis of results grouped by fentanyl use status was completed for an open‐label study with rapid induction of extended‐release buprenorphine in the inpatient research unit. Eligible participants received a single 4 mg dose of transmucosal buprenorphine (BUP‐TM) followed by an extended‐release buprenorphine 300 mg injection ([BUP‐XR]) after approximately 1 h. An extension study continued follow‐up up to 6 months (6 monthly injections).

Results

Among participants with fentanyl‐positive urine samples (FEN+; n = 19), all received BUP‐TM, 17 received BUP‐XR, 13 elected to receive a second BUP‐XR injection, and 10 received all six scheduled injections. Among participants with fentanyl‐negative samples (FEN−; n = 7), all received BUP‐TM and BUP‐XR, four elected to receive a second injection, and two participants received all six scheduled injections. Induction day clinical opioid withdrawal scale (COWS) scores were similar for FEN+ and FEN− groups. In the FEN+ group, mean COWS scores fell to below 5 within 24 h of BUP‐XR injection.

Discussion and Conclusions

The treatment of individuals with OUD using fentanyl with a rapid 1‐day induction to BUP‐XR 300 mg injection is feasible and well‐tolerated.

Scientific Significance

A prospective trial of participants grouped by fentanyl use status at induction demonstrates comparable patient retention and clinical response following single‐day induction of BUP‐XR in participants who are FEN+ and FEN−.
Intrauterine perfusion of dexamethasone improves pregnancy outcomes in recurrent reproductive failure patients with elevated uterine natural killer cells. A retrospective cohort study
Authors: Zhang Feng , Wang Zhuran , Lian Ruochun , Diao Lianghui , Li Yuye , Wu Yaya , Yin Tailang , Huang Chunyu
Published: November 2023

Objective

To determine the effect of intrauterine perfusion of dexamethasone (DXM) on pregnancy outcomes in recurrent reproductive failure (RRF) patients with elevated uNK cells.

Methods

This retrospective cohort study included 132 RRF patients with elevated uNK cells: 56 patients received DXM treatment and 76 patients refused it in the frozen‐thawed embryo transfer cycles. To determine the efficacy of intrauterine perfusion of DXM, multivariate logistic regression models and diagnosis‐based subgroup analysis were performed. We also compared the pregnancy outcomes of patients with different responsiveness to DXM treatment.

Results

Intrauterine perfusion of DXM significantly improved clinical pregnancy rate (aOR: 3.188, 95% CI: 1.395–7.282, P = .006) and live birth rate (aOR: 3.176, 95% CI: 1.318–7.656, P = .010) in RRF patients with elevated uNK cells, but there was no significant association with miscarriage rate. Subgroup analysis revealed that intrauterine perfusion of DXM in patients with recurrent implantation failure (RIF) showed significant improvement in clinical pregnancy rate (aOR: 6.110, 95% CI: 1.511–24.713, P = .011) and live birth rate (aOR: 9.904, 95% CI: 1.963–49.968, P = .005), but there was insufficient evidence of benefit in recurrent pregnancy loss (RPL) patients. Additionally, uNK cell levels dropped to normal range was achieved in only 35.90% of RRF patients after DXM treatment, no significant difference was found in pregnancy outcomes among patients with different responsiveness to DXM treatment (all P > .05).

Conclusion

Intrauterine perfusion of DXM was a promising and effective treatment to enhance clinical pregnancy rate and live birth rate in RRF women with abnormally elevated uNK cells, and RIF patients are more likely to benefit than RPL patients.