bluuebunny/crossref_metadata_2025 huggingface dataset:
Suppression of sost/sclerostin and dickkopf-1 promote intervertebral disc structure in mice
Authors: Kroon Tori , Niziolek Paul , Edwards Daniel , Clinkenbeard Erica L , Robling Alexander , Holguin Nilsson
Published: July 2021

Intervertebral disc (IVD) degeneration is a leading cause of low back pain and characterized by accelerated extracellular matrix breakdown and IVD height loss but there is no approved pharmacological therapeutic. Deletion of Wnt signaling receptor Lrp5 induces IVD degeneration and suggests that Wnt signaling in the IVD may be responsive to inhibition of Wnt signaling inhibitors sost(gene)/sclerostin(protein) or dickkopf-1 (dkk1). Anti-sclerostin antibody (Scl-Ab) is an FDA-approved bone therapeutic that activates Wnt signaling. We (1) determined if pharmacological neutralization of sclerostin, dkk1 or their combination stimulate Wnt signaling and promote IVD structure and (2) determined the extent of the response of the IVD to global, persistent deletion of sost. Nine-week-old C57Bl/6J female mice (n=6-7/grp) were subcutaneously injected 2x/wk for 5.5 wk with scl-Ab (25 mg/kg), dkk1-Ab (25 mg/kg), 3:1 scl-Ab/dkk1-Ab (18.75:6.25 mg/kg) or vehicle (Veh). Separately, IVD of sost KO and WT (wildtype) mice (n=8, grp) were harvested at 16 weeks of age. First, compared to vehicle, scl-Ab, dkk1-Ab and 3:1 scl-Ab/dkk1-Ab similarly increased lumbar IVD height and β-catenin gene expression. Despite these similarities, scl-Ab decreased cellular stress-related heat shock protein gene expressions while neither dkk1-Ab nor scl-Ab/dkk1-Ab altered the same. Genetically and compared to WT, sost KO increased MRI-determined hydration and proteoglycan staining in the IVD. Notably, persistent deletion of sost was compensated by upregulation of dkk1, which consequently reduced the cell nuclear fraction for Wnt signaling transcription factor β-catenin in whole IVD. Lastly, RNA-sequencing pathway analysis confirmed the compensatory suppression of Wnt signaling and determined a reduction of cellular stress pathways. Together, suppression of sost/sclerostin or dkk1 each promote IVD structure by stimulating Wnt signaling, but sclerostin and dkk1 may differentially regulate cellular stress pathways. Ultimately, postmenopausal women prescribed scl-Ab injections to prevent vertebral fracture may also benefit from a restoration of IVD height and health.

Graphical abstract

Suppression of Wnt signaling inhibitors by genetic or pharmacological approaches promotes intervertebral disc structure and hydration by Wnt signaling. However, persistent activation of Wnt signaling induces a compensatory reduction of Wnt signaling that shifts IVD cells toward a chondrocyte-like (CLC) phenotype. AF: annulus fibrosus, NC: notochordal cell, NP: nucleus pulposus, PG: proteoglycan
Mining all publicly available expression data to compute dynamic microbial transcriptional regulatory networks
Authors: Sastry Anand V. , Poudel Saugat , Rychel Kevin , Yoo Reo , Lamoureux Cameron R. , Chauhan Siddharth , Haiman Zachary B. , Al Bulushi Tahani , Seif Yara , Palsson Bernhard O.
Published: July 2021

We are firmly in the era of biological big data. Millions of omics datasets are publicly accessible and can be employed to support scientific research or build a holistic view of an organism. Here, we introduce a workflow that converts all public gene expression data for a microbe into a dynamic representation of the organism’s transcriptional regulatory network. This five-step process walks researchers through the mining, processing, curation, analysis, and characterization of all available expression data, usingBacillus subtilisas an example. The resulting reconstruction of theB. subtilisregulatory network can be leveraged to predict new regulons and analyze datasets in the context of all published data. The results are hosted athttps://imodulondb.org/, and additional analyses can be performed using the PyModulon Python package. As the number of publicly available datasets increases, this pipeline will be applicable to a wide range of microbial pathogens and cell factories.
Gaussian accelerated molecular dynamics reveals that a proline-rich signaling peptide frequently samples cis conformations when unbound
Authors: Alcantara Juan , Stix Robyn , Huang Katherine , Connor Acadia , East Ray , Jaramillo-Martinez Valeria , Stollar Elliott J. , Ball K. Aurelia
Published: July 2021

Disordered proline-rich motifs are common across the proteomes of many species and are often involved in protein-protein interactions. Proline is a unique amino acid due to the covalent bond between the backbone nitrogen and the proline side chain. The resulting five-membered ring allows proline to sample the cis state about its peptide bond, which other residues cannot do as readily. Because proline-rich disordered sequences exist as ensembles that likely include structures with the proline peptide bond in cis, a robust methodology to accurately account for these conformations in the overall ensemble is crucial. Observing the cis conformations of proline in a disordered sequence is challenging both experimentally and computationally. Nitrogen-hydrogen NMR spectroscopy cannot directly observe proline residues, which lack an amide bond, and computational methods struggle to overcome the large kinetic barrier between the cis and trans states, since isomerization usually occurs on the order of seconds. In the current work, Gaussian accelerated molecular dynamics was used to overcome this free energy barrier and simulate proline isomerization in a tetrapeptide (KPTP) and in the 12-residue proline-rich SH3 binding peptide, ArkA. We found that Gaussian accelerated molecular dynamics, when combined with a lowered peptide bond dihedral angle potential energy barrier (15 kcal/mol), allowed sufficient sampling of the proline cis and trans states on a microsecond timescale. All ArkA prolines spend a significant fraction of time in cis, leading to a more compact ensemble with less polyproline II helix structure than an ArkA ensemble with all peptide bonds in trans. The ensemble containing cis prolines also matches more closely to in vitro circular dichroism data than the all-trans ensemble. The ability of the ArkA prolines to isomerize likely affects the peptide’s ability to bind its partner SH3 domain, and should be studied further. This is the first molecular dynamics simulation study of proline isomerization in a biologically relevant proline-rich sequence that we know of, and a similar protocol could be applied to study multi-proline isomerization in other proline-containing proteins to improve conformational diversity and agreement with in vitro data.
Evolution of the quorum sensing regulon in cooperating populations of Pseudomonas aeruginosa
Authors: Smalley Nicole E. , Schaefer Amy L. , Asfahl Kyle L. , Perez Crystal , Greenberg E. Peter , Dandekar Ajai A.
Published: July 2021

The bacterium Pseudomonas aeruginosa is an opportunistic pathogen and it thrives in many different saprophytic habitats. In this bacterium acyl-homoserine lactone quorum sensing (QS) can activate expression of over 100 genes, many of which code for extracellular products. P. aeruginosa has become a model for studies of cell-cell communication and coordination of cooperative activities. We hypothesized that long-term growth of bacteria under conditions where only limited QS-controlled functions were required would result in a reduction in the size of the QS-controlled regulon. To test this hypothesis, we grew P. aeruginosa for about 1000 generations in a condition in which expression of QS-activated genes is required for growth. We compared the QS regulons of populations after about 35 generations to those after about 1000 generations in two independent lineages by using quorum quenching and RNA-seq technology. In one evolved lineage the number of QS-activated genes identified was reduced by about 70% and in the other by about 45%. Our results lend important insights about the variations in the number of QS-activated genes reported for different bacterial strains and, more broadly, about the environmental histories of P. aeruginosa.
OpenSense: An open-source toolbox for Inertial-Measurement-Unit-based measurement of lower extremity kinematics over long durations
Authors: Al Borno Mazen , O’Day Johanna , Ibarra Vanessa , Dunne James , Seth Ajay , Habib Ayman , Ong Carmichael , Hicks Jennifer , Uhlrich Scott , Delp Scott
Published: July 2021

Background

The ability to measure joint kinematics in natural environments over long durations using inertial measurement units (IMUs) could enable at-home monitoring and personalized treatment of neurological and musculoskeletal disorders. However, drift, or the accumulation of error over time, inhibits the accurate measurement of movement over long durations. We sought to develop an open-source workflow to estimate lower extremity joint kinematics from IMU data that was accurate, and capable of assessing and mitigating drift.

Methods

We computed IMU-based estimates of kinematics using sensor fusion and an inverse kinematics approach with a constrained biomechanical model. We measured kinematics for 11 subjects as they performed two 10-minute trials: walking and a repeated sequence of varied lower-extremity movements. To validate the approach, we compared the joint angles computed with IMU orientations to the joint angles computed from optical motion capture using root mean square (RMS) difference and Pearson correlations, and estimated drift using a linear regression on each subject’s RMS differences over time.

Results

IMU-based kinematic estimates agreed with optical motion capture; median RMS differences over all subjects and all minutes were between 3-6 degrees for all joint angles except hip rotation and correlation coefficients were moderate to strong (r = 0.60 to 0.87). We observed minimal drift in the RMS differences over ten minutes; the average slopes of the linear fits to these data were near zero (−0.14 to 0.17 deg/min).

Conclusions

Our workflow produced joint kinematics consistent with those estimated by optical motion capture, and could mitigate kinematic drift even in the trials of continuous walking without rest, obviating the need for explicit sensor recalibration (e.g. sitting or standing still for a few seconds or zero-velocity updates) used in current drift-mitigation approaches. This could enable long-duration measurements, bringing the field one step closer to estimating kinematics in natural environments.
Optimized nickase- and nuclease-based prime editing in human and mouse cells
Authors: Adikusuma Fatwa , Lushington Caleb , Arudkumar Jayshen , Godahewa Gelshan I. , Chey Yu C.J. , Gierus Luke , Piltz Sandra , Reti Daniel , Wilson Laurence OW. , Bauer Denis C. , Thomas Paul Q.
Published: July 2021

Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. Currently, the delivery of PE components to mammalian cell lines requires multiple plasmid vectors. To overcome this limitation, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embryonic stem (ES) cells. We discovered that PE efficiency in HEK293T cells was much higher than previously observed, reaching up to 95% (mean 67%). The efficiency in K562 and HeLa cells, however, remained low. To improve PE efficiency in K562 and HeLa, we generated a nuclease prime editor and tested this system in these cell lines as well as mouse ES cells. PE-nuclease greatly increased prime editing initiation, however, installation of the intended edits was often accompanied by extra insertions derived from the repair template. Finally, we show that zygotic injection of the nuclease prime editor can generate correct modifications in mouse fetuses with up to 100% efficiency. In summary, PE-nuclease and the PEA1 plasmids provide new tools to generate intended edits with high efficiency.
Aspects of the Neurospora crassa sulfur starvation response are revealed by transcriptional profiling and DNA affinity purification sequencing
Authors: Huberman Lori B , Wu Vincent W. , Lee Juna , Daum Chris , O'Malley Ronan , Glass N. Louise
Published: July 2021

Accurate nutrient sensing is important for rapid fungal growth and exploitation of available resources. Sulfur is an important nutrient source found in a number of biological macromolecules, including proteins and lipids. The model filamentous fungus Neurospora crassa is capable of utilizing sulfur found in a variety of sources from amino acids to sulfate. During sulfur starvation, the transcription factor CYS-3 is responsible for upregulation of genes involved in sulfur uptake and assimilation. Using a combination of RNA sequencing and DNA affinity purification sequencing, we performed a global survey of the N. crassa sulfur starvation response and the role of CYS-3 in regulating sulfur responsive genes. Along with genes known to be involved in sulfur metabolism, the CYS-3 transcription factor also directly activated the expression of a number of uncharacterized transporter genes, suggesting that regulating sulfur import is an important aspect of regulation by CYS-3. Additionally, CYS-3 directly regulated the expression of genes involved in mitochondrial electron transfer. During sulfur starvation, genes involved in nitrogen metabolism, such as amino acid and nucleic acid metabolic pathways, along with genes encoding proteases and nucleases that are necessary for scavenging nitrogen, were activated. Sulfur starvation also caused changes in the expression of genes involved in carbohydrate metabolism, such as those encoding glycosyl hydrolases. Thus, our data suggest a connection between sulfur metabolism and other aspects of cellular metabolism.
Associations between Acoustic Features of Maternal Speech and Infants’ Emotion Regulation following a Social Stressor
Authors: Kolacz Jacek , daSilva Elizabeth B. , Lewis Gregory F. , Bertenthal Bennett I. , Porges Stephen W.
Published: July 2021

Caregiver voices may provide cues to mobilize or calm infants. This study examined whether maternal prosody predicted changes in infants’ biobehavioral state during the Still Face, a stressor in which the mother withdraws and reinstates social engagement. Ninety-four dyads participated in the study (infant age 4-8 months). Infants’ heart rate and respiratory sinus arrhythmia (measuring cardiac vagal tone) were derived from an electrocardiogram (ECG). Infants’ behavioral distress was measured by negative vocalizations, facial expressions, and gaze aversion. Mothers’ vocalizations were measured with spectral analysis and spectro-temporal modulation using a two-dimensional fast Fourier transformation of the audio spectrogram. High values on the maternal prosody composite were associated with decreases in infants’ heart rate (β=-.26, 95% CI: [-.46, -.05]) and behavioral distress (β=-.20, 95% CI: [-.38, -.02]), and increases in cardiac vagal tone in infants whose vagal tone was low during the stressor (1 SD below mean β=.39, 95% CI: [.06, .73]). High infant heart rate predicted increases in the maternal prosody composite (β=.18, 95% CI: [.03, .33]). These results suggest specific vocal acoustic features of speech that are relevant for regulating infants’ biobehavioral state and demonstrate mother-infant bi-directional dynamics.
Functional constraints on insect immune system components govern their evolutionary trajectories
Authors: Ruzzante Livio , Feron Romain , Reijnders Maarten J.M.F. , Thiébaut Antonin , Waterhouse Robert M.
Published: July 2021

The role of constraints in shaping evolutionary outcomes is often investigated in the contexts of developmental biology and population genetics, in terms of the capacity to generate new variants as well as how selection either limits or promotes consequent phenotypic change. Comparative genomics also recognises the role of constraints, in terms of shaping the evolution of gene and genome architectures, sequence evolutionary rates, and gene gains and losses, as well as on molecular phenotypes. Characterising patterns of genomic change where putative functions and interactions of system components are relatively well-described offers opportunities to explore whether genes with similar or analogous roles exhibit similar evolutionary trajectories, possibly governed by common constraints. Using insect innate immunity as our study system, we hypothesise that quantitative characterisation of gene evolutionary histories can define distinct dynamics associated with different functional roles. We develop metrics that quantify gene evolutionary histories, employ these to characterise evolutionary features of immune gene repertoires, and explore relationships between gene family evolutionary profiles and their roles in immunity to understand how different constraints may relate to distinct dynamics. We identified three main axes of evolutionary trajectories characterised by gene duplication and synteny, maintenance/stability and sequence conservation, and loss and sequence divergence, highlighting similar and contrasting patterns across these axes amongst subsets of immune genes. Our results indicate that where and how genes participate in immune responses limit the range of possible evolutionary scenarios they exhibit. Comparative genomics approaches therefore offer opportunities to characterise how functional constraints on different components of biological systems govern their evolutionary trajectories.
Neutrophil and natural killer cell imbalances prevent muscle stem cell mediated regeneration following murine volumetric muscle loss
Authors: Larouche Jacqueline A. , Kurpiers Sarah J. , Yang Benjamin A. , Davis Carol , Fraczek Paula M. , Hall Matthew , Brooks Susan V. , Shea Lonnie D. , Aguilar Carlos A.
Published: July 2021

Volumetric muscle loss (VML) overwhelms the innate regenerative capacity of mammalian skeletal muscle (SkM), leading to numerous disabilities and reduced quality of life. Immune cells are critical responders to muscle injury and guide tissue resident stem cell and progenitor mediated myogenic repair. However, how immune cell infiltration and inter-cellular communication networks with muscle stem cells are altered following VML and drive pathological outcomes remains underexplored. Herein, we contrast the cellular and molecular mechanisms of VML injuries that result in fibrotic degeneration or regeneration of SkM. Following degenerative VML injuries, we observe heightened infiltration of natural killer (NK) cells as well as persistence of neutrophils beyond two weeks post injury. Functional validation of NK cells revealed an antagonistic role on neutrophil accumulation in part via inducing apoptosis and CCR1 mediated chemotaxis. The persistent infiltration of neutrophils in degenerative VML injuries was found to contribute to impairments in muscle stem cell regenerative function, which was also attenuated by transforming growth factor beta 1 (TGFβ1). BlockingTGFβsignaling reduced neutrophil accumulation and fibrosis, as well as improved muscle specific force. Collectively, these results enhance our understanding of immune cell-stem cell crosstalk that drives regenerative dysfunction and provide further insight into possible avenues for fibrotic therapy exploration.

SINGLE SENTENCE SUMMARY

Comparison of muscle injuries resulting in regeneration or fibrosis reveals inter-cellular communication between neutrophils and natural killer cells impacts muscle stem cell mediated repair.
A validation standard for Area of Habitat maps for terrestrial birds and mammals
Authors: Dahal Prabhat R. , Lumbierres Maria , Butchart Stuart H. M. , Donald Paul F. , Rondinini Carlo
Published: July 2021

Area of Habitat (AOH) is a deductive model which maps the distribution of suitable habitat at suitable altitudes for a species inside its broad geographical range. AOH maps have been validated using presence-only data for small subsets of species for different taxonomic groups, but no standard validation method exists when absence data are not available. We develop a novel two-step validation protocol for AOH which includes first a model-based evaluation of model prevalence (i.e, the proportion of suitable habitat within a species’ range), and second a validation using species point localities (presence-only) data. We applied the protocol to AOH maps of terrestrial birds and mammals. In the first step we built logistic regression models to predict expected model prevalence (the proportion of the range retained as AOH) as a function of each species’ elevation range, mid-point of elevation range, number of habitats, realm and, for birds, seasonality. AOH maps with large difference between observed and predicted model prevalence were identified as outliers and used to identify a number of sources of systematic error which were then corrected when possible. For the corrected AOH, only 1.7% of AOH maps for birds and 2.3% of AOH maps for mammals were flagged as outliers in terms of the difference between their observed and predicted model prevalence. In the second step we calculated point prevalence, the proportion of point localities of a species falling in pixels coded as suitable in the AOH map. We used 48,336,141 point localities for 4889 bird species and 107,061 point localities for 420 mammals. Where point prevalence exceeded model prevalence, the AOH was a better reflection of species’ distribution than random. We also found that 4689 out of 4889 (95.9%) AOH maps for birds, and 399 out of 420 (95.0%) AOH maps for mammals were better than random. Possible reasons for the poor performance of a small proportion of AOH maps are discussed.
Light and electron microscopy continuum-resolution imaging of 3D cell cultures
Authors: D’Imprima Edoardo , Garcia Montero Marta , Gawrzak Sylwia , Ronchi Paolo , Zagoriy Ievgeniia , Schwab Yannick , Jechlinger Martin , Mahamid Julia
Published: July 2021

3D cell cultures, in particular organoids, are emerging models to investigate healthy or diseased tissues. Understanding the complex cellular sociology in organoids requires integration of imaging modalities across spatial and temporal scales. We present a multi-scale imaging approach that traverses millimeter-scale live-cell light microscopy to nano-scale volume electron microscopy by performing 3D cell cultures in a single carrier amenable to all imaging steps. This allows to follow organoids growth, probe their morphology with fluorescent markers, identify areas of interest and analyze their 3D ultrastructure. We demonstrate this workflow on mouse and human 3D cultures, and use automated image segmentation to annotate and quantitatively analyze subcellular structures in patient-derived colorectal cancer organoids. Our analyses reveal local organization of diffraction-limited cell junctions in compact and polarized epithelia. The continuum resolution imaging pipeline is thus suited to foster basic and translational organoid research by simultaneously exploiting the advantages of light and electron microscopy.

Highlights

Establishment of 3D cell cultures in sample carriers directly amenable to high-pressure freezing (HPF)

3D cell cultures in HPF carriers allow drug treatment and live-cell imaging

Multi-scale imaging of 3D cultures from live-cell light microscopy to volume electron microscopy

Establishments of HPF conditions for mouse and patient-derived organoids

Deep-learning automatic segmentation of ultrastructural detail and quantitative data-mining reveal different subcellular organization associated with epithelium polarity
Mitochondrial dysfunction triggers secretion of the immunosuppressive factor α-fetoprotein
Authors: Jett Kimberly A. , Baker Zakery N. , Hossain Amzad , Boulet Aren , Cobine Paul A. , Ghosh Sagnika , Ng Philip , Yilmaz Orhan , Barreto Kris , DeCoteau John , Mochoruk Karen , Ioannou George N. , Savard Christopher , Yuan Sai , Lowden Christopher , Kim Byung-Eun , Cheng Hai-Ying Mary , Battersby Brendan J. , Gohil Vishal M. , Leary Scot C.
Published: July 2021

Signaling circuits crucial to systemic physiology are widespread, yet uncovering their molecular underpinnings remains a barrier to understanding the etiology of many metabolic disorders. Here, we identify a copper-linked signaling circuit activated by disruption of mitochondrial function in the murine liver or heart that results in atrophy of the spleen and thymus and causes a peripheral white blood cell deficiency. We demonstrate that the leukopenia is caused by α-fetoprotein, which requires copper and the cell surface receptor CCR5 to promote white blood cell death. We further show that α-fetoprotein expression is upregulated in several cell types upon inhibition of oxidative phosphorylation, including a muscle cell model of Barth syndrome. Collectively, our data argue that α-fetoprotein secreted by bioenergetically stressed tissue suppresses the immune system, an effect which may explain the recurrent infections that are observed in a subset of mitochondrial diseases or in other disorders with mitochondrial involvement.
The number of cytokinesis nodes in mitotic fission yeast scales with cell size
Authors: Sayyad Wasim A. , Pollard Thomas D.
Published: July 2021

Cytokinesis nodes are assemblies of stoichiometric ratios of proteins associated with the plasma membrane, which serve as precursors for the contractile ring during cytokinesis by fission yeast. The total number of nodes is uncertain, because of the limitations of the methods used previously. Here we used the ∼140 nm resolution of Airyscan confocal microscopy to resolve a large population of small, unitary cytokinesis nodes in 3D reconstructions of whole fission yeast cells. Wild-type fission yeast cells make about 200 unitary cytokinesis nodes early in mitosis. Most, but not all of these nodes condense into a contractile ring. The number of cytokinesis nodes scales with cell size in four strains tested, although wide rga4Δ mutant cells form somewhat fewer cytokinesis nodes than expected from the overall trend. The surface density of Pom1 kinase on the plasma membrane around the equators of cells is similar with a wide range of node numbers, so Pom1 does not control cytokinesis node number. However, varying protein concentrations with the nmt1 promoter showed that the numbers of nodes increase above a baseline of about 200 with the total cellular concentration of either kinase Pom1 or Cdr2.
Socioeconomic determinants of primary school children dropout: a case study of Pakistan
Authors: Ali Kazim , Yaseen Muhammad Rizwan , Makhdum Muhammad Sohail Amjad , Quddoos Abdul , Sardar Azeem
Published: July 2021

Purpose

The main purpose of this study is to identify the socioeconomic determinants of dropout from primary schools and to give policy suggestions to address the issue.

Design/methodology/approach

A total of 600 dropout and enrolled respondents were selected from 60 government primary schools of district Chiniot. School heads and parents of dropout children were taken as samples. The results were obtained by employing the Probit regression model.

Findings

Numbers of family members, age of the family head, exchange marriage and poverty status have positive relationship with dropout from primary schools. The findings revealed a higher rate of dropout among girls, which is a major cause of concern.

Practical implications

Education is regarded as a basic human right and a valuable human capital. It is included in Millennium Development Goals to achieve universal primary education and in Sustainable Development Goals as quality education. Underdeveloped countries are facing the problems of high dropout and lack of quality education, especially in Pakistan. These problems need to be addressed to keep pace with developed nations and to meet development goals.

Originality/value

It is recommended that government should create employment opportunities, family planning programs, legislature measures on exchange marriage and child labor. The involvement in co-curricular activities in learning and usage of audio-visual aids in the teaching process can improve the enrollment in the primary schools.
Growth inhibition of phytopathogenic spiroplasmas by membrane‐interactive antimicrobial peptides Novispirin T7 and Caerin 1.1
Authors: Wei Wei , Davis Robert E. , Mowery Joseph D. , Zhao Yan
Published: July 2021

Spiroplasma kunkelii and Spiroplasma citri, both helical‐shaped cell wall‐less bacteria, are the causative agents of corn stunt disease and citrus stubborn disease, respectively. Plants exhibiting natural resistance to these phytopathogenic spiroplasmas are currently lacking. Engineering artificial plant resistance using antimicrobial peptides (AMPs) has been conceived as a new approach to control the agronomically important spiroplasmal diseases. In preparation for such task, the present study focused on screening of AMPs that have potentials to curb the growth of S. kunkelii and S. citri. Four AMPs, including Novispirin T7, Caerin 1.1, Tricholongin and Dhvar4, were selected for in vitro growth inhibition test. A liquid assay method was developed for quick qualitative and quantitative evaluations of the AMPs. Our results demonstrated that Novispirin T7 and Caerin 1.1 were able to inhibit the growth of both phytopathogenic spiroplasmas with the efficacy comparable to that of tetracycline. Cell deformations were observed in spiroplasma cultures treated with these two peptides, indicating interactions of the AMPs with the spiroplasma cell membranes. The minimum inhibitory concentrations (MICs) of the AMPs against S. kunkelii and S. citri were determined.
Knowledge, views and expectations for cancer polygenic risk testing in clinical practice: A cross‐sectional survey of health professionals
Authors: Smit Amelia K. , Sharman Ashleigh R. , Espinoza David , Wallingford Courtney , Young Mary‐Anne , Dunlop Kate , Tiller Jane , Newson Ainsley J. , Meiser Bettina , Cust Anne E. , Yanes Tatiane
Published: July 2021

Polygenic risk scores (PRS) are becoming increasingly available in clinical practice to evaluate cancer risk. However, little is known about health professionals' knowledge, attitudes, and expectations of PRS. An online questionnaire was distributed by relevant health professional organisations predominately in Australia, Canada and the US to evaluate health professionals' knowledge, views and expectations of PRS. Eligible participants were health professionals who provide cancer risk assessments. Results from the questionnaire were analysed descriptively and content analysis was undertaken of free‐text responses. In total, 105 health professionals completed the questionnaire (genetic counsellors 84%; oncologists 6%; clinical geneticists 4%; other 7%). Although responses differed between countries, most participants (61%) had discussed PRS with patients, 20% had ordered a test and 14% had returned test results to a patient. Confidence and knowledge around interpreting PRS were low. Although 69% reported that polygenic testing will certainly or likely influence patient care in the future, most felt unprepared for this. If scaled up to the population, 49% expect that general practitioners would have a primary role in the provision of PRS, supported by genetic health professionals. These findings will inform the development of resources to support health professionals offering polygenic testing, currently and in the future.
Fish reproductive activity reveals temporal variations predominating spatial heterogeneity in maintaining high functional diversity of a Neotropical reservoir
Authors: Gogola Tatiane Mary , Piana Pitágoras Augusto , da Silva Pedro Rogério Leandro , Topan Dhyego Amilton , Sanches Paulo Vanderlei
Published: July 2021

The reproductive activity of the fish assemblage in a large Neotropical reservoir was evaluated by surveying the occurrence of ichthyoplankton in response to spatial and temporal variation in environmental factors. Fish reproduction was investigated by capturing larvae along representative spatial gradients in the main body (longitudinal) and arms (lateral) of the reservoir. After identification, the captured larvae were classified into different groups according to the functional traits of their parental stock. Sampling occurred monthly at night from October to March over five reproductive periods between 2009 and 2016, using conical‐cylindrical plankton nets with flowmeter. Water samples were taken simultaneously allowing the measurement of environmental factors related to reproductive activity. The relationship between the environmental factors and the functional traits was assessed using RLQ and fourth‐corner analyses. An evaluation of 51,995 larvae revealed there was greater larval abundance in the lacustrine and transition zones of both longitudinal and lateral gradients. Occurrences often correlated with temporal instead of spatial gradients. Higher rainfall correlated with abundances of detritivorous and migratory species. Higher water levels correlated with higher abundances of non‐native species, species without parental care, with external fecundation, and partial spawning. Dry periods, with lower water and higher dissolved oxygen, correlated with higher abundances of insectivores and sedentary species. Although the presence of a dam‐free stretch of river upstream of the reservoir had a positive influence on fish larvae in the fluvial zone, the results did not show differentiation along either gradient. Thus, the composition of the functional groups of fish species that reproduce in the Itaipu Reservoir appears to be predominantly influenced by the temporal variation in environmental factors rather than the formation of spatial gradients.
Association between serum secretory phospholipase A2 and risk of ischaemic stroke
Authors: Smith Jed W. , Barlas Raphae S. , Mamas Mamas A. , Boekholdt S. Matthijs , Mallat Ziad , Luben Robert N. , Wareham Nicholas J. , Khaw Kay‐Tee , Myint Phyo K.
Published: July 2021

Background and purpose

Previous literature has demonstrated an association between high serum levels of type II secretory phospholipase A2 (sPLA2) concentration and an increased risk of coronary artery disease. However, such association has not been established in terms of ischaemic stroke risk. The aim was to evaluate the association between both sPLA2 concentration and activity as continuous variables with risk of future ischaemic stroke.

Methods

A nested case–control study was conducted using data from the European Prospective Investigation into Cancer—Norfolk study. Cases (n = 145) in the current study were participants who developed ischaemic stroke during follow‐up, with controls (n = 290) matched in a 2:1 ratio based on age and sex. Statistical analyses were performed using SPSS (version 25.0) software. Logistic regression was used to determine odds ratios (OR) and corresponding 95% confidence intervals (95% CIs) for ischaemic stroke.

Results

After adjusting for a wide array of cardiovascular confounders, sPLA2 activity was found to be associated with an increased risk of ischaemic stroke using both multiple imputations with chained equations and complete case analysis: OR 1.20 (95% CI 1.01–1.43) and OR 1.23 (95% CI 1.01−1.49), respectively. However, sPLA2 concentration was not found to be associated with increased risk of ischaemic stroke.

Conclusions

The activity of sPLA2, but not sPLA2 concentration, is associated with an increased risk of future ischaemic stroke. This finding may be significant in risk group stratification, allowing targeted prophylactic treatment, or the development of novel therapeutic agents.
Tuning the luminescence properties of blue and far‐red dual emitting Gd2MgTiO6: Bi3+, Cr3+ phosphor for LED plant lamp
Authors: Gao Peixin , Zhou Zeyun , Dong Peng , Li Qian , Li Honghui , Wang Jing , Zhou Zhi , Xia Mao
Published: July 2021

Blue and far‐red light play a key role in plant growth, so it is necessary to develop blue and far‐red dual emitting phosphors. However, the match between phosphors and plant pigments is not satisfactory. In this work, we synthesized a series of blue and far‐red dual emission Gd2MgTiO6: Bi3+, Cr3+ (GMTO: Bi3+, Cr3+) phosphors and discussed the luminescence performance. The blue emission at 430 nm is ascribed to 3P1 → 1S0 transition of Bi3+ and the far‐red emission is ascribed to 4T2 → 4A2 and 2E → 4A2 transitions of Cr3+. Notably, because of the energy competition between Cr3+ ions and host materials, the luminescence tuning realized with the content of Cr3+ doping. In addition, an energy‐transfer performance occurred from Bi3+ ions to Cr3+ ions and the photoluminescence intensity of Cr3+ can be enhanced by Bi3+. The pc‐LEDs devices were synthesized by GMTO: Bi3+, Cr3+ phosphor, and ultraviolet (UV) chips. Finally, the emission of GMTO: Bi3+, Cr3+ phosphor matched well with the absorption spectra of plant pigments which indicated the potential applications in LED plant lamp.
Gender stereotypes and self‐characterizations in Germany and Nigeria: A cross‐cultural comparison
Authors: Obioma Ihuoma F. , Hentschel Tanja , Hernandez Bark Alina S.
Published: July 2021

Studies on the content of gender stereotypes have been conducted primarily in the United States, while research in other, particularly non‐Western, countries is scarce. In this research, we assessed and compared the content of gender stereotypes and self‐characterizations in Germany—a Western European country—and Nigeria—a West African country. We asked 403 Germans and Nigerians to rate three target groups (either men in general, women in general, or themselves) on 74 agentic and communal characteristics. We found that Nigerian women were rated as more agentic and more communal than German women, while German men were rated as more communal than Nigerian men, but similarly on agency. On self‐characterizations, Nigerian men rated themselves as more communal than German men, but again similarly on agency; Nigerian women rated themselves as more agentic and more communal than German women. Within‐country comparisons showed that in Germany, men compared to women were perceived as more agentic but similarly communal when rated by others (and similarly agentic and communal when rating themselves). In Nigeria, men and women were perceived as similarly agentic, but women were perceived as more communal than men (by both others and when rating themselves). Further analysis on individual agentic and communal characteristics, however, showed important differences in stereotypes and self‐characterizations of men and women in both countries that were obscured when looking at overall agency and communion. Our results show that gender stereotyping of oneself and others is complex and highlights the impact of culture on people's perceptions of gender.
Stepwise endo‐/epicardial catheter ablation for atrial fibrillation: The Mediterranea approach
Authors: De Martino Giuseppe , Compagnucci Paolo , Mancusi Carmine , Vassallo Enrico , Calvanese Claudia , Della Ratta Giuseppe , Librera Mariateresa , Franciulli Marco , Marino Luigi , Russo Antonio Dello , Casella Michela
Published: July 2021

Background

Outcomes of catheter ablation (CA) among patients with nonparoxysmal atrial fibrillation (AF) are largely disappointing.

Objective

We sought to evaluate the feasibility, effectiveness, and safety of a single‐stage stepwise endo‐/epicardial approach in patients with persistent/longstanding‐persistent AF.

Methods

We enrolled 25 consecutive patients with symptomatic persistent (n = 4) or longstanding‐persistent (n = 21) AF and at least one prior endocardial procedure, who underwent CA using an endo‐/epicardial approach. Our anatomical stepwise protocol included multiple endocardial as well as epicardial (Bachmann's bundle [BB] and ligament of Marshall ablations) components, and entailed ablation of atrial tachycardias emerging during the procedure. The primary outcome was freedom from any AF/atrial tachycardia episode after a 3‐month blanking period. The secondary outcome was patients' symptom status during follow‐up.

Results

The stepwise endo‐/epicardial approach allowed sinus rhythm restoration in 72% of patients, either directly (n = 6, 24%) or after AF organization into atrial tachycardia (n = 12, 48%). BB's ablation was commonly implicated in arrhythmia termination. After a median follow‐up of 266 days (interquartile range, 96 days), survival free from AF/atrial tachycardia was 88%. Antiarrhythmic drugs could be discontinued in 22 patients (88%). As compared to baseline, more patients were asymptomatic at 9‐month follow‐up (0% vs. 56%, p = .02). Five patients (20%) developed mild medical complications, whereas one subject (4%) had severe kidney injury requiring dialysis.

Conclusion

A single‐stage endo‐/epicardial CA resulted in favorable rhythm and symptom outcomes in a cohort of patients with symptomatic persistent/longstanding‐persistent AF and one or more prior endocardial procedures. Epicardial ablation of BB was commonly implicated in procedural success.
Csnk1a1 inhibition modulates the inflammatory secretome and enhances response to radiotherapy in glioma
Authors: Liu Guanzheng , Li Huan , Zhang Wanhong , Yu Jiefeng , Zhang Xu , Wu Runqiu , Niu Mingshan , Liu Xuejiao , Yu Rutong
Published: July 2021

Glioblastoma multiforme (GBM), a fatal brain tumour with no available targeted therapies, has a poor prognosis. At present, radiotherapy is one of the main methods to treat glioma, but it leads to an obvious increase in inflammatory factors in the tumour microenvironment, especially IL‐6 and CXCL1, which plays a role in tumour to resistance radiotherapy and tumorigenesis. Casein kinase 1 alpha 1 (CK1α) (encoded on chromosome 5q by Csnk1a1) is considered an attractive target for Tp53 wild‐type acute myeloid leukaemia (AML) treatment. In this study, we evaluated the anti‐tumour effect of Csnk1a1 suppression in GBM cells in vitro and in vivo. We found that down‐regulation of Csnk1a1 or inhibition by D4476, a Csnk1a1 inhibitor, reduced GBM cell proliferation efficiently in both Tp53 wild‐type and Tp53‐mutant GBM cells. On the contrary, overexpression of Csnk1a1 promoted cell proliferation and colony formation. Csnk1a1 inhibition improved the sensitivity to radiotherapy. Furthermore, down‐regulation of Csnk1a1 reduced the production and secretion of pro‐inflammatory factors. In the preclinical GBM model, treatment with D4476 significantly inhibited the increase in pro‐inflammatory factors caused by radiotherapy and improved radiotherapy sensitivity, thus inhibiting tumour growth and prolonging animal survival time. These results suggest targeting Csnk1a1 exert an anti‐tumour role as an inhibitor of inflammatory factors, providing a new strategy for the treatment of glioma.
Effects of therapeutic clowning on pain and anxiety during venous blood sampling in Turkey: Randomised controlled trial
Authors: Kurudirek Fatma , Arikan Duygu , Sarialioğlu Arzu
Published: July 2021

Purpose

The research aim was to investigate the effects of therapeutic clowning on pain and fear during the process of venous blood sampling in children.

Design and Methods

This study was conducted in a randomised controlled trial from January to March 2020. The study population consisted of children from 7 to 12 years of age who came to the research centre for blood sampling. The research sample consisted of 166 children (83 in the intervention group and 83 in the control group) who met the research criteria and came to the research centre on a specific date through probable sampling. For the data collection, a questionnaire form, the Visual Analogue Scale and the Children's Fear Scale were used. Therapeutic clowning was used in this process for the intervention group. There was no intervention for children who were in the control group. The calculation of percentage distribution and means, χ2 test, analysis of variance analysis in repeated measures and least significant difference and independent t‐test were used to analyse the data.

Results

The results of the research indicated that children in the intervention group had significantly lower pain and fear than children in the control group (p < .001). Therapeutic clowning seems to effectively reduce pain and fear during venous blood sampling for children from 7 to 12 years of age. Our results support using therapeutic clowns which is an inexpensive, safe, and effective method for lowering children's pain and fear during venous blood sampling.

Practice Implications

The use of therapeutic clowns could become the standard of care in blood sampling for children from 7 to 12 years of age in hospitals. Therapeutic clowning, which is an easy‐to‐use, low‐cost and practical method to provide better communication and cooperation with the family and the child, can be used as a complementary therapy in all nursing areas, especially in the field of paediatrics. Nurses could be encouraged to be trained on the use of therapeutic humour based upon a standardised approach and offered guidance on how to apply entertaining methods such as clowns. Therapeutic clowning can be examined in other applications in nursing care in light of these results.
Vagueness and social ontology: Implications of inquiry resistant borderline cases for social ontological theorising
Authors: Meyenburg Imko , Turcitu Ana Maria
Published: July 2021

The aim of this paper is to introduce the philosophical concept of vagueness and use it to critique the explanatory scope of social ontology as developed by the Cambridge Social Ontology Group (CSOG), and scholars sympathetic to it. Specifically, we will refer to one of its core theories, the theory of social positioning, to formulate our critique. This theory proposes that human beings and artefacts occupy social positions within emerging social totalities by virtue of receiving community accepted, interdependent, rights and obligations. Vagueness, here, is to be understood as a matter of indeterminacy in borderline cases that pose non‐trivial limitations to social positioning. By this we mean to say that there are instances in social ontological theorising that exclude themselves from proper inquiry due to vague terms found in natural languages. As a consequence of these vague terms, theories in social ontology formulated in such languages face limitations when striving for precisification and accuracy of theories. We further argue that these limitations cannot be overcome, but introduce four theories, namely supervaluationsim, epistemicism, truth degree theories and contextualism, which will allow interested readers to understand why these limitations are present. Finally, we consider the controversial theory of ontic vagueness and outline the implications for social ontology.
Forest cover at landscape scales increases male and female gametic diversity of palm seedlings
Authors: Diaz‐Martin Zoe , Karubian Jordan
Published: July 2021

Genetic diversity shapes the evolutionary potential of plant populations. For outcrossing plants, genetic diversity is influenced by effective population size and by dispersal, first of paternal gametes through pollen, and then of paternal and maternal gametes through seeds. Forest loss often reduces genetic diversity, but the degree to which it differentially impacts the paternal and maternal contributions to genetic diversity and the spatial scale at which these impacts are most pronounced are poorly understood. To address these questions, we genotyped 504 seedlings of the animal‐dispersed palm Oenocarpus bataua collected from 29 widely distributed sites across Ecuador and decomposed the contribution of paternal and maternal gametes to overall genetic diversity. The amount of forest cover at a landscape scale (>10 km radius) had an equally significant positive association with both male and female gametic diversity. In addition, there was a significant positive association between forest cover and effective population size. Stronger fine‐scale spatial genetic structure for female versus male gametes was observed at sites with low forest cover, but this did not scale up to differences in male versus female gametic diversity. These findings show that reductions in forest cover at spatial scales much larger than those typically evaluated in ecological studies lead to significant, and equivalent, decreases of diversity in both male and female gametes, and that this association between landscape level forest loss and genetic diversity may be driven directly by reductions in effective population size of O. bataua, rather than by indirect disruptions to local dispersal processes.