bluuebunny/crossref_metadata_2025 huggingface dataset:
Real-time dynamics of individual chemoreceptor mRNA molecules reveals translation hotspots at the inner membrane ofEscherichia coli
Authors: Bergmiller Tobias , Krasnopeeva Ekaterina , Sarikas Srdjan , Nikolic Nela , Guet Calin C.
Published: December 2022

Since bacteria lack a nucleus, the location of mRNA molecules is determined by the different characteristics of the encoded proteins, and the transcriptome is spatially arranged into cytosolic and membrane-associated mRNA. While translation of membrane protein-encoding mRNA has been studied in great mechanistic detail using biochemical methods, the spatiotemporal dynamics of this process remains poorly understood at the subcellular level. Here, we investigate the dynamics of individual fluorescently labelled mRNA molecules encoding the transmembrane serine chemoreceptor Tsr, to probe the mechanism of membrane protein translation. Analysis oftsrmRNA diffusion in the proximity of the plasma membrane revealed distinct diffusive modes that reflect the state of the mRNA molecule and its involvement in the process of active translation into the Sec secretion system. We find that the composition, and hence the fluidity of the membrane affects diffusion of membrane targeted mRNAs. Moreover, Tsr translation occurs in localized membrane regions, similar to eukaryotic hotspots. The hotspot localization coincides with the physical location of the transcribed gene, which itself is displaced towards the inner membrane. These findings suggest that inner membrane protein translation is a spatially defined process that occurs in hotspots, indicative of long-lived transertion sites. Our results show an additional layer of spatio-temporal structuring within the bacterial cell, thus revealing a qualitatively different understanding of the basic process of transcription and translation in bacteria.

Significance Statement

A large fraction of the bacterial proteome is directly synthesized into the inner membrane, and this process shapes the overall distribution of mRNA transcripts within the cell. Although highly dynamic in their nature, bacterial transcriptomes have mostly been studied in fixed cells. Here, we track individual mRNA molecules encoding the serine chemoreceptor in living bacterial cells and find that translation occurs in membrane hotspots that were previously exclusive to eukaryotes. Our results indicate an additional layer of spatio-temporal structuring within the bacterial cell that impacts our understanding of transcription and translation in bacteria.
Scaling the Functional Nanopore (FuN) Screen: Systematic Evaluation of Self-Assembling Membrane Peptides and Extension with a K+-Responsive Fluorescent Protein Sensor
Authors: Eisenhauer Klara , Weber Wadim , Kemp Philipp , Gebhardt Carolin , Kaufmann Marwan , Tewes Noel , Zhadanowa Hanna , Tietze Alesia , Rauh Oliver , Stein Viktor
Published: December 2022

The functional analysis of protein nanopores is typically conducted in planar lipid bilayers or liposomes exploiting high-resolution but low-throughput electrical and optical read-outs. Yet, the reconstitution of protein nanoporesin vitrostill constitutes an empiric and low-throughput process. Addressing these limitations, nanopores can now be analyzed using the functional nanopore (FuN) screen exploiting genetically-encoded fluorescent protein sensors that resolve distinct nanopore-dependent Ca2+in- and efflux patterns across the inner membrane ofEscherichia coli. With a primary proof-of-concept established for the S2168 holin, and thereof based recombinant nanopore assemblies, the question arises to what extent alternative nanopores can be analyzed with the FuN screen and to what extent alternative fluorescent protein sensors can be adapted. Focussing on self-assembling membrane peptides, three sets of 13 different nanopores are assessed for their capacity to form nanopores in the context of the FuN screen. Nanopores tested comprise both natural and computationally-designed nanopores. Further, the FuN screen is extended to K+-specific fluorescent protein sensors to provide a complementary read-out to Ca2+. Finally, a comparison with high-resolution biophysical and electrophysiological studies in planar lipid bilayers provides an experimental benchmark for future studies.
Morphogenesis of bullet-shaped rabies virus particles requires a functional interplay between the viral matrix protein and ESCRT-I component TSG101
Authors: Itakura Yukari , Tabata Koshiro , Saito Takeshi , Intaruck Kittiya , Kawaguchi Nijiho , Kishimoto Mai , Torii Shiho , Kobayashi Shintaro , Ito Naoto , Harada Michiko , Inoue Satoshi , Maeda Ken , Takada Ayato , Hall William W. , Orba Yasuko , Sawa Hirofumi , Sasaki Michihito
Published: December 2022

Viral protein assembly and virion budding are tightly regulated to enable the proper formation of progeny virions. At this late stage in the virus life cycle, some enveloped viruses take advantage of the host ESCRT (endosomal sorting complex required for transport) machinery, which contributes to the physiological functions of membrane modulation and abscission. Bullet-shaped viral particles are unique morphological characteristics of rhabdoviruses; however, the involvement of host factors in rhabdovirus infection, and specifically the molecular mechanisms underlying virion formation are not fully understood. In the present study, we used a siRNA screening approach and found that the ESCRT-I component TSG101 contributes to the propagation of rabies virus (RABV). We demonstrated that the matrix protein (M) of RABV interacts with TSG101 via the late-domain containing the PY and YL motifs, which are conserved in various viral proteins. Loss of the YL motif in the RABV M or the downregulation of host TSG101 expression resulted in the intracellular aggregation of viral proteins and abnormal virus particle formation, indicating a defect in the RABV assembly and budding processes. These results indicate that the interaction of the RABV M and TSG101 is pivotal for not only the efficient budding of progeny RABV from infected cells but also for the bullet-shaped virion morphology.

Importance

Enveloped-viruses bud from cells with the host lipid bilayer. Generally, the membrane modulation and abscission are mediated by host ESCRT (endosomal sorting complex required for transport) complexes. Some enveloped-viruses utilize their late (L)-domain to interact with ESCRTs, which promotes viral budding. Rhabdoviruses form characteristic bullet-shaped enveloped-virions, but the underlying molecular mechanisms involved remain elusive. Herein, we showed that TSG101, one of ESCRT components, supports rabies virus (RABV) budding and proliferation. TSG101 interacted with RABV matrix protein via L-domain, and the absence of this interaction resulted in intracellular virion accumulation and distortion of the morphology of progeny virions. Our study reveals that virion formation of RABV is highly regulated by TSG101 and the virus matrix protein.
Different modification pathways for m1A58 incorporation in yeast elongator and initiator tRNAs
Authors: Yared Marcel-Joseph , Yoluç Yasemin , Catala Marjorie , Tisné Carine , Kaiser Stefanie , Barraud Pierre
Published: December 2022

As essential components of the cellular protein synthesis machineries, tRNAs undergo a tightly controlled biogenesis process, which include the incorporation of a large number of posttranscriptional chemical modifications. Maturation defaults resulting in lack of modifications in the tRNA core may lead to the degradation of hypomodified tRNAs by the rapid tRNA decay (RTD) and nuclear surveillance pathways. Although modifications are typically introduced in tRNAs independently of each other, several modification circuits have been identified in which one or more modifications stimulate or repress the incorporation of others. We previously identified m1A58 as a late modification introduced after more initial modifications, such as Ѱ55 and T54 in yeast elongator tRNAPhe. However, previous reports suggested that m1A58 is introduced early along the tRNA modification process, with m1A58 being introduced on initial transcripts of initiator tRNAiMet, and hence preventing its degradation by the nuclear surveillance and RTD pathways. Here, aiming to reconcile this apparent inconsistency on the temporality of m1A58 incorporation, we examined the m1A58 modification pathways in yeast elongator and initiator tRNAs. For that, we first implemented a generic approach enabling the preparation of tRNAs containing specific modifications. We then used these specifically modified tRNAs to demonstrate that the incorporation of T54 in tRNAPheis directly stimulated by Ѱ55, and that the incorporation of m1A58 is directly and individually stimulated by Ѱ55 and T54, thereby reporting on the molecular aspects controlling the Ѱ55 → T54 → m1A58 modification circuit in yeast elongator tRNAs. We also show that m1A58 is efficiently introduced on unmodified tRNAiMet, and does not depend on prior modifications. Finally, we show that the m1A58 single modification has tremendous effects on the structural properties of yeast tRNAiMet, with the tRNA elbow structure being properly assembled only when this modification is present. This rationalizes on structural grounds the degradation of hypomodified tRNAiMetlacking m1A58 by the nuclear surveillance and RTD pathways.
An optimized and validated workflow for developing stable producer cell lines with >99.99% assurance of clonality and high clone recovery
Authors: Scherzinger Julia , Türk Daniel , Aprile-Garcia Fernando
Published: December 2022

There is a constant pressure to reduce timelines in mammalian cell line development (CLD) for biotherapeutic protein production. Demonstration of clonal derivation of the generated cell lines is key for health authorities’ approval. To meet these regulatory and process-oriented demands, single-cell dispensers have become vital instruments for single-cell cloning. We conducted validation experiments with the UP.SIGHT (CYTENA GmbH) to determine this instrument’s single-cell dispensing efficiency (SCDE) and probability of clonal derivation (p(clonal)). Process optimization to maximize clone recovery with several cell lines was also performed, focusing on cloning media and plate type. With a SCDE >97%, p(clonal) >99.99% and clone recovery values of up to 80%, the data reported here support the notion that the UP.SIGHT covers all steps in the single-cell dispensing process with assurance of clonality and colony tracking, leading to faster and more efficient CLD workflows. This work also serves as a guideline for instrument validation and guidance towards process optimization.
ClpP/ClpX deficiency impairs mitochondrial functions and mTORC1 signaling during spermatogenesis and meiosis
Authors: Guo Chenxi , Xiao Yuan , Gu Jingkai , Hu Zhe , Zheng Jiahuan , Hua Renwu , Hai Zhuo , Su Jiaping , Zhang Jian V. , Yeung William S.B. , Wang Tianren
Published: December 2022

Caseinolytic protease proteolytic subunit (ClpP) and caseinolytic protease X (ClpX) are mitochondrial matrix peptidases that activate mitochondrial unfolded protein response to maintain protein homeostasis in the mitochondria. However, the role of ClpP and ClpX in spermatogenesis remains largely unknown. In this study, we demonstrated the importance of ClpP/ClpX for meiosis and spermatogenesis with two conditional knockout (cKO) mouse models. We found that ClpP/ClpX deficiency reduced mitochondrial functions and quantity in spermatocytes, affected energy supply during meiosis and attenuated zygotene-pachytene transformation of the male germ cells. The dysregulated spermatocytes finally underwent apoptosis resulting in decreased testicular size and vacuolar structures within the seminiferous tubules. We found mTORC1 pathway was over-activated after deletion of ClpP/ClpX in spermatocytes. Long-term inhibition of the mTORC1 signaling via rapamycin treatmentin vivopartially rescue spermatogenesis. The data reveal the novel roles of ClpP and ClpX in regulating meiosis and spermatogenesis.

One-Sentence Summary

ClpP/ClpX is required for maintaining mitochondrial functions in spermatocytes during meiosis and spermatogenesis.
Distinct molecular profile of the chick Organizer as a stem zone during axial elongation
Authors: Wood Timothy R. , Kucinski Iwo , Voiculescu Octavian
Published: December 2022

The vertebrate Organizer plays a crucial role in defining the main axes of embryo: it neuralizes the surrounding ectoderm, and is the site of emigration for cells making axial and paraxial mesendoderm during elongation. The chick Organizer becomes a stem zone at the onset of elongation: it stops recruiting cells from the neighbouring ectoderm, and generates all its derivatives from the small number of resident cells it contains at the end of gastrulation stages. Nothing is known about the molecular identity of this stem zone. Here, we specifically labelled long-term resident cells of the Organizer, and compared their RNA-seq profile to that of the neighbouring cell populations. Screening by RT-PCR and in situ hybridisation identified four genes (WIF1, PTGDS, ThPOandUCKL1), which are upregulated only in the Organizer region when it becomes a stem zone, and remain expressed there during axial elongation. In experiments specifically labelling the resident cells of the mature Organizer, we show that only these cells express these genes. These findings molecularly define the Organizer as a stem zone, and offer a key to understanding how this zone is set up, the molecular control of its cells’ behaviour, and the evolution of axial growth zones.
Bumblebees actively compensate for the adverse effects of sidewind during visually-guided landings
Authors: Goyal Pulkit , van Leeuwen Johan L. , Muijres Florian T.
Published: December 2022

Flying animals often encounter winds during visually guided landings. However, how winds affect their flight control strategy during landing is unknown. Here, we investigated how sidewind affects the landing strategy, sensorimotor control, and landing performance of foraging bumblebees (Bombus terrestris). For this, we trained a hive of bumblebees to forage in a wind tunnel, and used high-speed stereoscopic videography to record 19,421 landing flight maneuvers in six sidewind speeds (0 to 3.4 m s−1), which correspond to winds encountered in nature. Bumblebees landed less often in higher windspeeds, but the landing duration from free flight was not increased by wind. We then tested how bumblebees adjusted their landing control to compensate for the adverse effects of sidewind on landing. This showed that the landing strategy in sidewind was similar to that in still air, but with important adaptations. In the highest windspeeds, more hover phases occurred than during landings in still air. The rising hover frequency did not increase landing duration because bumblebees flew faster in between hover phases. Hence, they negated the adverse effects of increased hovering in high windspeeds. Using control theory, we revealed how bumblebees integrate information from the wind-mediated mechanosensory modality with their vision-based sensorimotor control loop. The proposed multi-sensory flight control system may be commonly used by insects landing in windy conditions and it may inspire the development of landing control strategies onboard man-made flying systems.

Summary statement

Bumblebees foraging in strong sidewinds can still land precisely on artificial flowers, allowing them to be efficient and robust pollinators in these adverse environmental conditions.
Tuning in Sensorimotor Synchronization
Authors: Michalareas Georgios , Grabenhorst Matthias , Sun Yue
Published: December 2022

Moving in synchrony to external rhythmic stimuli is an elementary function that humans regularly engage in. It is termed “sensorimotor synchronization” and it is governed by two main parameters, the period and the phase of the movement with respect to the external rhythm. There has been an extensive body of research on the characteristics of these parameters, primarily once the movement synchronization has reached a steady-state level. Particular interest has been shown about how these parameters are corrected when there are deviations for the steady-state level. However, little is known about the initial “tuning-in” interval, when one aligns the movement to the external rhythm from rest. The current work investigates this “tuning-in” period for each of the four limbs and makes various novel contributions in the understanding of sensorimotor synchronization. The results suggest that phase and period alignment appear to be separate processes. Phase alignment involves limb-specific somatosensory memory in the order of minutes while period alignment has very limited memory usage. Phase alignment is the primary task but then the brain switches to period alignment where it spends most its resources. In overall this work suggests a central, cognitive role of period alignment and a peripheral, sensorimotor role of phase alignment.

Highlights

In the tuning-in phase there are three distinct temporal scales of sensorimotor synchronization with distinct signatures. A long-range, across-blocks monotonic negative gradient to more anticipatory movement, which prevails for tens of minutes, a very consistent “hook”-shaped pattern within each block, in the range of seconds, and a constant difference across time between feet and hands.

The across-blocks, monotonic, negative gradient to more anticipatory movement is instantiated only in the first anticipatory trial of each block and the rest of the subsequent block trials contribute to the alignment of the inter-movement interval to the metronome’s period.

This negative asynchrony gradient is limb-specific and is not affected by the interleaved blocks of other limbs.

Period alignment has a central, cognitive role while phase alignment a peripheral, sensorimotor role.
Ligand Gaussian accelerated molecular dynamics 2 (LiGaMD2): Improved calculations of ligand binding thermodynamics and kinetics with closed protein pocket
Authors: Wang Jinan , Miao Yinglong
Published: December 2022

Ligand binding thermodynamics and kinetics are critical parameters for drug design. However, it has proven challenging to efficiently predict ligand binding thermodynamics and kinetics from molecular simulations due to limited simulation timescales. Protein dynamics especially in the ligand binding pocket often plays an important role in ligand binding. Based on our previously developed Ligand Gaussian accelerated molecular dynamics (LiGaMD), here we present LiGaMD2 in which a selective boost potential was applied to both the ligand and protein residues in the binding pocket to improve sampling of ligand binding and dissociation. To validate the performance of LiGaMD2, the T4 lysozyme (T4L) mutants with open and closed pockets bound by different ligands were chosen as model systems. LiGaMD2 could efficiently capture repetitive ligand dissociation and binding within microsecond simulations of all T4L systems. The obtained ligand binding kinetic rates and free energies agreed well with available experimental values and previous modeling results. Therefore, LiGaMD2 provides an improved approach to sample opening of closed protein pockets for ligand dissociation and binding, thereby allowing for efficient calculations of ligand binding thermodynamics and kinetics.
The Timing of Cortical Activation in Associator Grapheme-Colour Synaesthetes using MEG
Authors: Michalareas Giorgos , Kusnir Flor , Thut Gregor , Gross Joachim
Published: December 2022

Grapheme-colour synaesthetes experience an anomalous form of perception in which graphemes systematically induce specific colour concurrents in their mind’s eye (“associator” type). Although grapheme-colour synaesthesia has been well characterized behaviourally, its neural mechanisms remain largely unresolved. There are currently several competing models, which can primarily be distinguished according to the anatomical and temporal predictions of synaesthesia-inducing neural activity. The first main model (Cross-Activation/ Cascaded Cross-Tuning) positsearlyrecruitment of occipital colour areas in the initial feed-forward sweep of brain activity. The second (Disinhibited Feedback) posits: (i) later involvement of a multisensory convergence zone (for example, in parietal cortices) after graphemes have been processed in their entirety; and (ii) subsequent feedback to early visual areas (i.e., occipital colour areas). In this study, we examine both the timing and anatomical correlates of associator grapheme-colour synaesthetes (n=6) using MEG. Using unbiased analysis methods with littlea prioriassumptions, we applied Independent Component Analysis (ICA) on a single-subject level to identify the dominant patterns of the induced, synaesthetic percept. We observed evoked activity that significantly dissociates between synaesthesia-inducing and non-inducing conditions at approximately 190 ms following grapheme presentation. This effect is present in grapheme-colour synaesthetes, but not in matched controls, and exhibits an occipito-parietal topology localised consistently across individuals to extrastriate visual cortices and to superior parietal lobes. Due to the observed timing of this evoked activity and its localisation, our results support a model predicting relatively late synaesthesia-inducing activity, more akin to the Disinhibited Feedback model. In light of previous findings, our study suggest that differential timing and neural mechanisms may account for associator grapheme-colour synaesthetes, as compared to projectors.

Highlights

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first MEG study to-date examining associator grapheme-colour synaesthetes

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we use single-subj. ICA to dissociate synaesthesia-inducing vs. non-inducing activity

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we observe evoked activity ∼190 ms following grapheme presentation

effects localize consistently to extrastriate visual and superior parietal cortices

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the observed timing is consistent with a model of synaesthesia supporting relatively late activity of synaesthesia-inducing stimuli

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our findings suggest that differential mechanisms may underlie different sub-types of grapheme-colour synaesthesia
Surface-modified measles vaccines encoding oligomeric, fusion-stabilized SARS-CoV-2 spike glycoproteins bypass measles seropositivity, boosting neutralizing antibody responses to omicron and historical variants
Authors: Muñoz-Alía Miguel Á. , Nace Rebecca A. , Balakrishnan Baskar , Zhang Lianwen , Packiriswamy Nandakumar , Singh Gagandeep , Warang Prajakta , Mena Ignacio , Narjari Riya , Vandergaast Rianna , García-Sastre Adolfo , Schotsaert Michael , Russell Stephen J.
Published: December 2022

Serum titers of SARS-CoV-2 neutralizing antibodies (nAb) correlate well with protection from symptomatic COVID-19, but decay rapidly in the months following vaccination or infection. In contrast, measles-protective nAb titers are life-long after measles vaccination, possibly due to persistence of the live-attenuated virus in lymphoid tissues. We therefore sought to generate a live recombinant measles vaccine capable of driving high SARS-CoV-2 nAb responses. Since previous clinical testing of a live measles vaccine encoding a SARS-CoV-2 spike glycoprotein resulted in suboptimal anti-spike antibody titers, our new vectors were designed to encode prefusion-stabilized SARS-CoV-2 spike glycoproteins, trimerized via an inserted peptide domain and displayed on a dodecahedral miniferritin scaffold. Additionally, to circumvent the blunting of vaccine efficacy by preformed anti-measles antibodies, we extensively modified the measles surface glycoproteins. Comprehensivein vivomouse testing demonstrated potent induction of high titer nAb in measles-immune mice and confirmed the significant incremental contributions to overall potency afforded by prefusion stabilization, trimerization, and miniferritin-display of the SARS-CoV-2 spike glycoprotein, and vaccine resurfacing. In animals primed and boosted with a MeV vaccine encoding the ancestral SARS-CoV-2 spike, high titer nAb responses against ancestral virus strains were only weakly cross-reactive with the omicron variant. However, in primed animals that were boosted with a MeV vaccine encoding the omicron BA.1 spike, antibody titers to both ancestral and omicron strains were robustly elevated and the passive transfer of serum from these animals protected K18-ACE2 mice from infection and morbidity after exposure to BA.1 and WA1/2020 strains. Our results demonstrate that antigen engineering can enable the development of potent measles-based SARS-CoV-2 vaccine candidates.
Initial social-policy responses to the COVID-19 pandemic in the Global North – A scoping review
Authors: Mäntyneva Päivi , Ketonen Eeva-Leena , Hiilamo Heikki
Published: December 2022

Purpose

The purpose of this scoping review is to analyse comparative studies on social-policy measures implemented during the COVID-19 pandemic in Global North welfare states. The authors also consider the potential influence of the regimes on the responses.

Design/methodology/approach

The authors conducted a scoping review of six databases including peer-reviewed comparative studies. In an iterative process with exact inclusion criteria, the authors screened 699 titles/abstracts/articles and found 16 comparative research articles to be included in the review and analysis. The review summarises the main themes of the comparative articles and the articles' typical features.

Findings

The results show that social-policy measures were directed specifically at working-age people to minimise income loss and to save jobs. The pandemic also increased care-related responsibilities, necessitating the expansion of current policies and the implementation of new instruments. Despite the differences in responses between universalistic and residual welfare states, the influence of welfare regimes on COVID-19 social-policy measures remains unclear. The emergency responses in the different regimes varied widely in terms of coverage.

Research limitations/implications

The results of this review provide a basis on which to conduct future studies, identify new research topics and knowledge gaps and inspire new research questions and hypotheses. Given the accumulation of scientific knowledge in the area of social-policy measures, the need for systematic reviews will grow in the future.

Originality/value

The authors identified three main themes: changes in employment protection, changes in care-related income protection and the potential influence of welfare-state regimes on COVID-19-related measures.
Experimental extensification of mountain grasslands restores plant species richness but not species composition in the mid‐term
Authors: Humann‐Guilleminot Ségolène , Boch Steffen , Martínez‐De León Gerard , Lessard‐Therrien Malie , Hayoz‐Andrey Aline , Serres‐Hänni Alberto , Humbert Jean‐Yves
Published: December 2022

The traditional grasslands that characterize the cultural landscapes of the Palaeartic mountain massifs represent biodiversity hotspots. Yet, they are currently threatened by the intensification of farming practices, notably excesses in fertilization and irrigation.

We experimentally investigated the passive restoration of montane and subalpine hay meadows after 6 years of management intensification, with different levels of fertilization and irrigation, followed by 5 years of release of intensive management, that is, extensification. More specifically, relying on a full randomized block‐design replicated at 11 Swiss study sites constituted of extensively managed meadows, we exposed during 6 years (2010–2015) four 20 m diameter plots to three levels of intensification (low, medium and high inputs), while a fourth plot served as a control (no inputs). In the second phase of the experiment (2016–2020), all study meadows underwent farming extensification.

We monitored total species richness and plant diversity (Simpson diversity), indicator plant species as well as the composition and variability of the plant communities based on Bray–Curtis dissimilarity distances.

We found that total species richness decreased in the most intensified plots after 6 years of intensification, but all plots retrieved their baseline species richness after 5 years of re‐extensification. In addition, we found no difference between the years in plant diversity (Simpson diversity) among the treatments. Yet, intensification led to different plants communities' compositions in all three levels of intensification in 2015 compared to the extensive plots, and this structural difference remained after 5 years of re‐extensification.

Synthesis and applications. Land‐use intensification induces a rapid impoverishment of the flora of mountain meadows. Our results demonstrate the potential of mountain hay meadows to passively restore plant species richness after re‐extensification; however, plants communities did not fully recover. We recommend maintaining fertilization inputs as low as possible and operating active restoration on grasslands formerly intensified.
The diversity and distribution of introduced plant species reflect 8000 years of settlement history
Authors: Macek Martin , Abraham Vojtěch , Tkáč Peter , Novák David , Kolář Jan
Published: December 2022

Human population has affected natural ecosystems since prehistoric times in many ways, causing disturbances in existing ecosystems and creating novel habitats, and altering the colonisation and extinction rates with potentially long‐lasting effects on biodiversity. Here, we explored the pervasive effects of past human occupancy on present‐day diversity and the distribution of plant species introduced by humans in the distant past—archaeophytes—at the regional spatial scale.

We analysed spatial relations between the present‐day species richness of archaeophytes and native flora, the environmental setting, archaeological evidence, and the relationship between the residence time of archaeophytes and their regional range size. We used fine‐scaled gridded information on plant diversity and archaeological records for the period 6000 BCE to 1000 CE summarised as average occupancy probability (AOP) in Czechia, Central Europe.

The proportion of archaeophytes in the local flora positively correlated with AOP. Variation partitioning revealed largely overlapping effects of AOP, environmental conditions, and present‐day land use on the relative diversity of archaeophytes in local flora. The relationship between the minimum residence time of introduced species and their regional range size was non‐significant.

Synthesis. Present‐day regional diversity of archaeophytes mirrored the intensity of past human settlements. Our results suggest that the main underlying mechanism is the environmental filtering of non‐native species pools, while dispersal limitation plays a minor role in the regional patterns of archaeophyte diversity.
The MocR family transcriptional regulator DnfR has multiple binding sites and regulates Dirammox gene transcription in Alcaligenes faecalis JQ135
Authors: Xu Si‐Qiong , Wang Xiao , Xu Lu , Wang Ke‐Xin , Jiang Yin‐Hu , Zhang Fu‐Yin , Hong Qing , He Jian , Liu Shuang‐Jiang , Qiu Ji‐Guo
Published: December 2022

Microbial ammonia oxidation is vital to the nitrogen cycle. A biological process, called Dirammox (direct ammonia oxidation, NH3→NH2OH→N2), has been recently identified in Alcaligenes ammonioxydans and Alcaligenes faecalis. However, its transcriptional regulatory mechanism has not yet been fully elucidated. The present study characterized a new MocR‐like transcription factor DnfR that is involved in the Dirammox process in A. faecalis strain JQ135. The entire dnf cluster was composed of 10 genes and transcribed as five transcriptional units, that is, dnfIH, dnfR, dnfG, dnfABCDE and dnfF. DnfR activates the transcription of dnfIH, dnfG and dnfABCDE genes, and represses its own transcription. The intact 1506‐bp dnfR gene was required for activation of Dirammox. Electrophoretic mobility shift assays and DNase I footprinting analyses showed that DnfR has one binding site in the dnfH‐dnfR intergenic region and two binding sites in the dnfG‐dnfA intergenic region. Three binding sites of DnfR shared a 6‐bp repeated conserved sequence 5′‐GGTCTG‐N17‐GGTCTG‐3′ which was essential for the transcription of downstream target genes. Cysteine and glutamate act as possible effectors of DnfR to activate the transcription of transcriptional units of dnfG and dnfABCDE, respectively. This study provided new insights in the transcriptional regulation mechanism of Dirammox by DnfR in A. faecalis JQ135.
Seduction, deception and technology
Authors: Tyler Susan
Published: December 2022

This paper proposes that exploring the use of internet pornography creates a potentially beneficial, albeit defensive liminal space that can be used therapeutically. The content of compelling sexual scenes can be viewed as the psyche’s way of mastering internal trauma and masking an inner emptiness. However, from a Jungian perspective, the use of internet pornography can also be seen as a patient’s distorted way of trying to make dissociated affects more bearable by triumphing over them. The sexual use of technology can express intrapsychic conflict encapsulated in what Stoller called ‘microdots’. Stoller’s microdots are unique scripts – a kind of emotional code that gives clinicians an opportunity to reveal unconscious processes and transcend them through meaning making. Wood’s work at the Portman Clinic on the chosen ‘compelling scenarios’ of pornography‐addicted patients is also examined with this Jungian lens. Jung’s idea of the transcendent function is used to show the value of holding the tension of fantasy and frustration provoked by intimacy. The paper illustrates how they come together within the therapeutic relationship to stimulate something new.
Projecting the fiscal impact of immigration in the European Union
Authors: Christl Michael , Bélanger Alain , Conte Alessandra , Mazza Jacopo , Narazani Edlira
Published: December 2022

The increasing flow of immigrants into Europe over the last decade has generated a range of considerations in the policy agenda of many receiving countries. One of the main considerations for policymakers and public opinion alike is whether immigrants contribute their ‘fair’ share to their host country's tax and welfare system. In this paper, we assess the net fiscal impact of intra‐EU and extra‐EU migration in 27 European Union (EU) Member States. We find that migrants in the EU, on average, contribute more than natives to welfare states. However, when we take an age‐specific life‐cycle perspective, we find that natives generally show a higher net fiscal contribution than both groups of migrants. Among migrants, extra‐EU migrants contribute less than intra‐EU migrants. We then use a demographic microsimulation model to project the potential net fiscal impact of migration in the EU into the future. We show that despite the fact that intra‐EU migration contributes to reduce the strong negative impact of population ageing, its contribution is not sufficient to offset the negative fiscal consequences.
Medicare beneficiaries with more comprehensive primary care physicians report better primary care
Authors: O'Malley Ann S. , Rich Eugene C. , Ghosh Arkadipta , Palakal Maya , Rose Tyler , Swankoski Kaylyn , Peikes Deborah , McCall Nancy
Published: December 2022

Objective

To examine whether primary care physician (PCP) comprehensiveness is associated with Medicare beneficiaries' overall rating of care from their PCP and staff.

Data Sources

We linked Medicare claims with survey data from Medicare beneficiaries attributed to Comprehensive Primary Care Plus (CPC+) physicians and practices.

Study Design

We performed regression analyses of the associations between two claims‐based measures of PCP comprehensiveness in 2017 and beneficiaries' rating of care from their PCP and practice staff in 2018.

Data Collection/Extraction Methods

The analytic sample included 6228 beneficiaries cared for by 3898 PCPs. Regressions controlled for beneficiary, physician, practice, and market characteristics.

Principal Findings

Beneficiaries with more comprehensive PCPs rated care from their PCP and practice staff higher than did those with less comprehensive PCPs. For each comprehensiveness measure, beneficiaries whose PCP was in the 75th percentile were more likely than beneficiaries whose PCP was in the 25th percentile to rate their care highly (2 percentage point difference,p = 0.02).

Conclusions

Medicare beneficiaries with more comprehensive PCPs rate overall care from their PCPs and staff higher than those with less comprehensive PCPs.
Racialized economic segregation and potentially preventable hospitalizations among Medicaid/CHIP‐enrolled children
Authors: Smith Laura Barrie , O'Brien Claire , Kenney Genevieve M. , Tabb Loni Philip , Verdeflor Alaisha , Wei Keqin , Lynch Victoria , Waidmann Timothy
Published: December 2022

Objective

To examine geographic variation in preventable hospitalizations among Medicaid/CHIP‐enrolled children and to test the association between preventable hospitalizations and a novel measure of racialized economic segregation, which captures residential segregation within ZIP codes based on race and income simultaneously.

Data Sources

We supplement claims and enrollment data from the Transformed Medicaid Statistical Information System (T‐MSIS) representing over 12 million Medicaid/CHIP enrollees in 24 states with data from the Public Health Disparities Geocoding Project measuring racialized economic segregation.

Study Design

We measure preventable hospitalizations by ZIP code among children. We use logistic regression to estimate the association between ZIP code‐level measures of racialized economic segregation and preventable hospitalizations, controlling for sex, age, rurality, eligibility group, managed care plan type, and state.

Data Extraction Methods

We include children ages 0–17 continuously enrolled in Medicaid/CHIP throughout 2018. We use validated algorithms to identify preventable hospitalizations, which account for characteristics of the pediatric population and exclude children with certain underlying conditions.

Principal Findings

Preventable hospitalizations vary substantially across ZIP codes, and a quarter of ZIP codes have rates exceeding 150 hospitalizations per 100,000 Medicaid‐enrolled children per year. Preventable hospitalization rates vary significantly by level of racialized economic segregation: children living in the ZIP codes that have the highest concentration of low‐income, non‐Hispanic Black residents have adjusted rates of 181 per 100,000 children, compared to 110 per 100,000 for children in ZIP codes that have the highest concentration of high‐income, non‐Hispanic white residents (p < 0.01). This pattern is driven by asthma‐related preventable hospitalizations.

Conclusions

Medicaid‐enrolled children's risk of preventable hospitalizations depends on where they live, and children in economically and racially segregated neighborhoods—specifically those with higher concentrations of low‐income, non‐Hispanic Black residents—are at particularly high risk. It will be important to identify and implement Medicaid/CHIP and other policies that increase access to high‐quality preventive care and that address structural drivers of children's health inequities.
Wool keratin as a novel alternative protein: A comprehensive review of extraction, purification, nutrition, safety, and food applications
Authors: Giteru Stephen G. , Ramsey Derek H. , Hou Yakun , Cong Lei , Mohan Anand , Bekhit Alaa El‐Din Ahmed
Published: December 2022

The growing global population and lifestyle changes have increased the demand for specialized diets that require protein and other essential nutrients for humans. Recent technological advances have enabled the use of food bioresources treated as waste as additional sources of alternative proteins. Sheep wool is an inexpensive and readily available bioresource containing 95%–98% protein, making it an outstanding potential source of protein for food and biotechnological applications. The strong structure of wool and its indigestibility are the main hurdles to achieving its potential as an edible protein. Although various methods have been investigated for the hydrolysis of wool into keratin, only a few of these, such as sulfitolysis, oxidation, and enzymatic processes, have the potential to generate edible keratin. In vitro and in vivo cytotoxicity studies reported no cytotoxicity effects of extracted keratin, suggesting its potential for use as a high‐value protein ingredient that supports normal body functions. Keratin has a high cysteine content that can support healthy epithelia, glutathione synthesis, antioxidant functions, and skeletal muscle functions. With the recent spike in new keratin extraction methods, extensive long‐term investigations that examine prolonged exposure of keratin generated from these techniques in animal and human subjects are required to ascertain its safety. Food applications of wool could improve the ecological footprint of sheep farming and unlock the potential of a sustainable protein source that meets demands for ethical production of animal protein.
Implementing the equality, diversity, and inclusion agenda in multinational companies: A framework for the management of (linguistic) diversity
Authors: Ciuk Sylwia , Śliwa Martyna , Harzing Anne‐Wil
Published: December 2022

Advancing, both conceptually and practically, the equality, diversity, and inclusion (EDI) agenda, which is notoriously difficult to implement, this paper addresses the under‐researched area of global diversity management (GDM) in multinational companies (MNCs). Drawing on Harrison and Klein's (2007) conceptualisations of diversity (separation, variety, and disparity) and two core concepts (fluidity and reciprocity) that reflect recent developments in the EDI literature, we propose a two‐step framework for implementing the EDI agenda through GDM. We argue that to achieve inclusion, we first need to think differently about diversity and differences (i.e., view diversity in a positive light and recognise and appreciate differences as fluid), in order to act differently (i.e., promote reciprocal effort to leverage diversity). We illustrate our framework with the specific case of linguistic diversity, a diversity dimension that is particularly salient, but also often neglected in MNCs, and discuss the implications of the proposed framework for EDI theory as well as human resource management policies and practice.
Comparative study of phenolic compounds reveals a positive relationship between astringency and the phenolic composition in table grape varieties
Authors: Feng Jiao , Nieuwenhuizen Niels , Atkinson Ross , Wang Wu , Zeng Jingjue , Zheng Huan , Tao Jianmin
Published: December 2022

Phenolic compounds and their derivatives play a major role in the intensity and characteristics of grape (Vitis vinifera) astringency. The present study investigated the relationship between phenolic composition and astringency of six commercial table grape varieties (two of each white‐, red‐, and black‐skinned). Qualitative and quantitative liquid chromatography–mass spectrometry analysis was used to identify the variety‐specific phenolic profiles in the skins and total astringency intensity was assessed and described by a trained sensory panel. Thirty phenolic compounds were identified among the six varieties. Principal component analysis of the phenolic profiles revealed that the intensity of astringency of grape skin was positively correlated with catechin, epicatechin, epicatechin‐3‐O‐gallate, and proanthocyanidin dimers B1, B2, and B3. A further orthogonal partial least‐squares discrimination analysis of these compounds showed that catechin was the substance most strongly and positively correlated (R = 0.904) with grape skin astringency.

Practical Application

This study provided a better understanding of the relationships between phenolic composition and table grape astringency and highlighted a potential metabolic marker that could be used as a predictor for the complex astringency sensory attributes of table grape berries.
Let families decide: Barriers and enablers to participation in family‐assisted therapy for older people in transition care
Authors: Lawler Katherine , Taylor Nicholas F. , Shields Nora
Published: December 2022

Objective

To understand the barriers and enablers to participation in family‐assisted therapy for older people in Transition Care.

Methods

A qualitative study, underpinned by interpretive description, was conducted at two public health services in Melbourne, Australia. Participants included patients in Transition Care, or their family members, who either participated in or chose not to participate in a family‐assisted therapy trial. Semi‐structured interviews were conducted, transcribed verbatim and analysed thematically.

Results

Forty‐four participants were interviewed (17 patients and 27 family members). The unifying theme was to let families decide about participation in family‐assisted therapy. The unifying theme was illustrated by three subthemes. The first, what is possible for the family now, described practical considerations including geography, paid and unpaid work structure and commitments and the presence of fit and willing social networks. The second, what is important to the family now, recognised the role of family priorities in deciding. Physical rehabilitation and extra therapy were of high importance to some families. For others, emotional support or searching for a residential aged care bed were more important at the time. Finally, how the family functions described the complexity of relationships and family history that impacted the decision to participate.

Conclusions

The decision to participate in family‐assisted therapy is complex and is best made by patients and their families. Clinicians offering family‐assisted therapy are encouraged to avoid assuming what will or will not work for families and instead, to let families decide.
Study management strategies to optimize student‐ and trainee‐led collaborative research
Authors:   TASMAN Collaborative
Published: December 2022

The collaborative research model is a powerful approach to answering surgical research questions that empowers and inspires medical students and trainees. The Opioid PrEscRiptions and usage After Surgery (OPERAS) study is a student‐ and trainee‐led international multi‐centre prospective cohort study developed in Australia and Aotearoa New Zealand. In this article, we will discuss (i) how the OPERAS study was conceptualized and structured; (ii) the channels through which information and education were communicated to collaborators; (iii) how data was stored in a secure and user‐friendly fashion and (iv) the lessons learned and expected goals for the future. We aim to describe how collaborative research studies can be designed to support early career researchers to make valuable contributions to the literature.