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The transport of transition metal ions by members of the SLC11/NRAMP family constitutes a ubiquitous mechanism for the uptake of Fe2+ and Mn2+ across all kingdoms of life. Despite the strong conservation of the family, two of its branches have evolved a distinct substrate preference with one mediating Mg2+ uptake in prokaryotes and another the transport of Al3+ into plant cells. Our previous work on the SLC11 transporter from Eggerthella lenta revealed the basis for its Mg2+ selectivity (Ramanadane et al., 2022). Here, we have addressed the structural and functional properties of a putative Al3+ transporter from Setaria italica. We show that the protein transports diverse divalent metal ions and binds the trivalent ions Al3+ and Ga3+, which are both presumable substrates. Its cryo-electron microscopy (cryo-EM) structure displays an occluded conformation that is closer to an inward- than an outward-facing state, with a binding site that is remodeled to accommodate the increased charge density of its transported substrate.

Immunoglobulin loci-transgenic animals are widely used in antibody discovery and increasingly in vaccine response modelling. In this study, we phenotypically characterised B-cell populations from the Intelliselect® Transgenic mouse (Kymouse) demonstrating full B-cell development competence. Comparison of the naïve B-cell receptor (BCR) repertoires of Kymice BCRs naïve human and murine BCR repertoires revealed key differences in germline gene usage and junctional diversification. These differences result in Kymice having CDRH3 length and diversity intermediate between mice and humans. To compare the structural space explored by CDRH3s in each species repertoire, we used computational structure prediction to show that Kymouse naïve BCR repertoires are more human-like than mouse-like in their predicted distribution of CDRH3 shape. Our combined sequence and structural analysis indicates that the naïve Kymouse BCR repertoire is diverse with key similarities to human repertoires, while immunophenotyping confirms that selected naïve B-cells are able to go through complete development.

A central regulatory mechanism of iron homeostasis in humans involves ferroportin (FPN), the sole cellular iron exporter, and the peptide hormone hepcidin, which inhibits Fe2+ transport and induces internalization and degradation of FPN. Dysregulation of the FPN/hepcidin axis leads to diverse pathological conditions, and consequently, pharmacological compounds that inhibit FPN-mediated iron transport are of high clinical interest. Here, we describe the cryo-electron microscopy structures of human FPN in complex with synthetic nanobodies and vamifeport (VIT-2763), the first clinical-stage oral FPN inhibitor. Vamifeport competes with hepcidin for FPN binding and is currently in clinical development for β-thalassemia and sickle cell disease. The structures display two distinct conformations of FPN, representing outward-facing and occluded states of the transporter. The vamifeport site is located in the center of the protein, where the overlap with hepcidin interactions underlies the competitive relationship between the two molecules. The introduction of point mutations in the binding pocket of vamifeport reduces its affinity to FPN, emphasizing the relevance of the structural data. Together, our study reveals conformational rearrangements of FPN that are of potential relevance for transport, and it provides initial insight into the pharmacological targeting of this unique iron efflux transporter.

Theoretical accounts disagree on the role of dopamine in intertemporal choice and assume that dopamine either promotes delay of gratification by increasing the preference for larger rewards or that dopamine reduces patience by enhancing the sensitivity to waiting costs. Here, we reconcile these conflicting accounts by providing empirical support for a novel process model according to which dopamine contributes to two dissociable components of the decision process, evidence accumulation and starting bias. We re-analyzed a previously published data set where intertemporal decisions were made either under the D2 antagonist amisulpride or under placebo by fitting a hierarchical drift diffusion model that distinguishes between dopaminergic effects on the speed of evidence accumulation and the starting point of the accumulation process. Blocking dopaminergic neurotransmission not only strengthened the sensitivity to whether a reward is perceived as worth the delay costs during evidence accumulation (drift rate) but also attenuated the impact of waiting costs on the starting point of the evidence accumulation process (bias). In contrast, re-analyzing data from a D1 agonist study provided no evidence for a causal involvement of D1R activation in intertemporal choices. Taken together, our findings support a novel, process-based account of the role of dopamine for cost-benefit decision making, highlight the potential benefits of process-informed analyses, and advance our understanding of dopaminergic contributions to decision making., eLife, 12, ISSN:2050-084X

Although medical providers are a trusted vaccination information source for parents, they do not universally support vaccination. Complementary medicine (CM) providers are particularly likely to hold vaccine hesitant (VH) views, and VH parents often consult with them. Little research compares VH of parents and providers, and if and how each is associated with uptake of recommended childhood vaccines.We defined non-timely receipt as recommended vaccines given 1 month later than officially recommended, based on vaccination records. We administered versions of the Parent Attitudes about Childhood Vaccines (PACV) 5-item survey instrument to 1256 parents and their children's pediatricians (N = 112, 40 CM-oriented, 72 biomedical [not CM-oriented]) to identify moderately (PACV-score 5-6) and highly (PACV-score 7+) hesitant providers/parents. We obtained multivariable adjusted odds ratios to test relationships between parental VH and provider type/VH, and between non-timely receipt of selected childhood vaccines and parental VH and provider type/VH.No biomedical providers were VH, 9 CM providers were moderately VH, and 17 were highly VH. Parents seeing moderately and highly hesitant providers had adjusted odds ratio (AOR) for being VH = 6.6 (95% confidence interval (CI), 3.1-14.0) and AOR = 31.3 (95% CI 16.8-58.3), respectively. Across all vaccine uptake endpoints, children of moderately and highly hesitant parents had 1.9-3.8 and 7.1-12.3 higher odds of non-timely vaccination, and children seeing highly hesitant CM providers had 4.9-9.4 higher odds. Children seeing moderately hesitant CM providers had 3.3 higher odds of non-timely vaccination for the 1st dose of measles and 3.5 higher odds for 1st dose of polio/pertussis/tetanus.VH by both parents and providers each is associated with non-timely childhood vaccination. As VH parents are more likely to consult with VH providers, interventions aimed at increasing timely vaccination need to primarily target VH providers and their clients.

In Switzerland, the implementation of national health and prevention strategies is regulated at the cantonal level. To date, no in-depth inventory outlining the cantonal health activities and legal frameworks for influenza vaccination exists.To assess the activities and legal frameworks for influenza vaccination during the 2019/20 season across all 26 Swiss cantons.Survey using a structured closed-ended questionnaire.The activities' range across the cantons is broad from 11 activities in the canton Vaud to one in cantons Grisons and Jura. French/Italian-speaking cantons conduct 1.5 times more influenza vaccination activities on average than the German-speaking cantons. 23 of 26 cantons authorize influenza vaccination by pharmacists. All cantons allow nurses to vaccinate under the responsibility of a doctor.Influenza vaccination activities and vaccination-specific legal frameworks vary by canton and linguistic regions. Future vaccination strategies should consider concerted approaches to optimize their successful implementation.

Relief of ongoing pain is a potent motivator of behavior, directing actions to escape from or reduce potentially harmful stimuli. Whereas endogenous modulation of pain events is well characterized, relatively little is known about the modulation of pain relief and its corresponding neurochemical basis. Here, we studied pain modulation during a probabilistic relief-seeking task (a ‘wheel of fortune’ gambling task), in which people actively or passively received reduction of a tonic thermal pain stimulus. We found that relief perception was enhanced by active decisions and unpredictability, and greater in high novelty-seeking trait individuals, consistent with a model in which relief is tuned by its informational content. We then probed the roles of dopaminergic and opioidergic signaling, both of which are implicated in relief processing, by embedding the task in a double-blinded cross-over design with administration of the dopamine precursor levodopa and the opioid receptor antagonist naltrexone. We found that levodopa enhanced each of these information-specific aspects of relief modulation but no significant effects of the opioidergic manipulation. These results show that dopaminergic signaling has a key role in modulating the perception of pain relief to optimize motivation and behavior.

There is evidence for the effectiveness of youth combined plyometric and sprint with change of direction (COD) training. However, the evidence is not well-known regarding the in-season effects of biological age (peak height velocity—PHV) on the motor adaptive processes following combined plyometric and COD with ball training (P-CODBT) in youth soccer players. This study aimed to examine the in-season effects of P-CODBT (8 weeks and twice a week) on the athletic performances of male youth soccer players, circa- and post-PHV. In a randomized controlled training study with pre-to-post measurements, forty-eight male players were assigned into two experimental (performing P-CODBT; n = 12 × circa-PHV and n = 12 × post-PHV) and two control groups (CONG; n = 12 × circa-PHV and n = 12 × post-PHV). The pre- and post-training participants were assessed for their anthropometric, linear sprinting with and without a ball, COD speed with and without a ball, vertical jump, dynamic balance, and endurance-intensive performances. After the intervention, the experimental condition induced significant (all p < 0.0001) and small to large effect size (ES = 0.263–3.471) additional gains only on explosive measures compared to CONG. Both the experimental (all p < 0.0001; ES = 0.338–1.908) and control (p = 0.011–0.0001; ES = 0.2–1.8) groups improved their athletic performances over the training period. The improvements generated by p-CODBT were not affected by biological age. In-season short-term P-CODBT (twice a week) could be safe way to generate benefits in explosive performances in youth soccer players, which are relevant components of match-winning actions in soccer.

Spinally projecting serotonergic neurons play a key role in controlling pain sensitivity and can either increase or decrease nociception depending on physiological context. It is currently unknown how serotonergic neurons mediate these opposing effects. Utilizing virus-based strategies and Tph2-Cre transgenic mice, we identified two anatomically separated populations of serotonergic hindbrain neurons located in the lateral paragigantocellularis (LPGi) and the medial hindbrain, which respectively innervate the superficial and deep spinal dorsal horn and have contrasting effects on sensory perception. Our tracing experiments revealed that serotonergic neurons of the LPGi were much more susceptible to transduction with spinally injected AAV2retro vectors than medial hindbrain serotonergic neurons. Taking advantage of this difference, we employed intersectional chemogenetic approaches to demonstrate that activation of the LPGi serotonergic projections decreases thermal sensitivity, whereas activation of medial serotonergic neurons increases sensitivity to mechanical von Frey stimulation. Together these results suggest that there are functionally distinct classes of serotonergic hindbrain neurons that differ in their anatomical location in the hindbrain, their postsynaptic targets in the spinal cord, and their impact on nociceptive sensitivity. The LPGi neurons that give rise to rather global and bilateral projections throughout the rostrocaudal extent of the spinal cord appear to be ideally poised to contribute to widespread systemic pain control., eLife, 12, ISSN:2050-084X

This study aimed to demonstrate the changes of selected blood parameters in relation to 25(OH)D concentration during the autumn period in young soccer players. A total of 35 participants’ results (age: 17.5 ± 0.6 years, body mass 71.3 ± 6.9 kg) were tested twice: in mid-September and in mid-December and divided into subgroups with regard to two criteria. First, according to the initial level of the 25(OH)D concentration (optimal group—ODG, suboptimal group—SDG), second, according to drops in 25(OH)D concentration (high drop group—HDG, low drop group—LDG). A significant decrease (p < 0.001) in the 25(OH)D concentration was reported in the total group (TGr) and in all subgroups. Blood parameters such as white blood cells, red blood cells, haemoglobin and haematocrit increased significantly (p < 0.05) in TGr during the analysed period of time. The analysis of changes in the lipid profile did not expose significant differences except triglycerides. The asparagine amino transferase and creatine kinase activity decreased significantly after autumn in all analysed groups. The declining level of 25(OH)D concentration should be compensated (e.g., with vitamin D supplementation) during autumn. Applied training loads could also influence the blood parameters variability in young soccer players. Regular measurements of 25(OH)D concentration are helpful in identifying potential drops and allows for the preparation of individual supplementation plans for the players.

Peer reviewed: True, Tumour heterogeneity is thought to be a major barrier to successful cancer treatment due to the presence of drug resistant clonal lineages. However, identifying the characteristics of such lineages that underpin resistance to therapy has remained challenging. Here, we utilise clonal transcriptomics with WILD-seq; Wholistic Interrogation of Lineage Dynamics by sequencing, in mouse models of triple-negative breast cancer (TNBC) to understand response and resistance to therapy, including BET bromodomain inhibition and taxane-based chemotherapy. These analyses revealed oxidative stress protection by NRF2 as a major mechanism of taxane resistance and led to the discovery that our tumour models are collaterally sensitive to asparagine deprivation therapy using the clinical stage drug L-asparaginase after frontline treatment with docetaxel. In summary, clonal transcriptomics with WILD-seq identifies mechanisms of resistance to chemotherapy that are also operative in patients and pin points asparagine bioavailability as a druggable vulnerability of taxane-resistant lineages.

Simple Summary Spinal Cord Injury is a devastating condition that compromises the individual’s health, quality of life and functional independence. Rats submitted to Spinal Cord Injury were evaluated after four weeks of resistance training. Analyses of levels of muscle damage and oxidative stress surgery were performed. Resistance training demonstrated increase antioxidative activity while decreased oxidative damage in injured rats, in addition to having presented changes in the levels of muscle damage in that same group. The results highlight that resistance training promoted a decrease in oxidative stress and a significant response in muscle damage markers. Abstract Background: Spinal cord injury (SCI) is a condition that affects the central nervous system, is characterized by motor and sensory impairments, and impacts individuals’ lives. The objective of this study was to evaluate the effects of resistance training on oxidative stress and muscle damage in spinal cord injured rats. Methodology: Forty Wistar rats were selected and divided equally into five groups: Healthy Control (CON), Sham (SHAM) SCI Untrained group (SCI-U), SCI Trained group (SCI- T), SCI Active Trained group (SCI- AT). Animals in the trained groups were submitted to an incomplete SCI at T9. Thereafter, they performed a protocol of resistance training for four weeks. Results: Significant differences in muscle damage markers and oxidative stress in the trained groups, mainly in SCI- AT, were found. On the other hand, SCI- U group presented higher levels of oxidative stress and biomarkers of LDH and AST. Conclusion: The results highlight that resistance training promoted a decrease in oxidative stress and a significative response in muscle damage markers.

Funder: Donation from the Estate of Dr Hans Salvisberg, Funder: Nomis Foundation; Id: http://dx.doi.org/10.13039/501100008483, A defining characteristic of mammalian prions is their capacity for self-sustained propagation. Theoretical considerations and experimental evidence suggest that prion propagation is modulated by cell-autonomous and non-autonomous modifiers. Using a novel quantitative phospholipase protection assay (QUIPPER) for high-throughput prion measurements, we performed an arrayed genome-wide RNA interference (RNAi) screen aimed at detecting cellular host-factors that can modify prion propagation. We exposed prion-infected cells in high-density microplates to 35,364 ternary pools of 52,746 siRNAs targeting 17,582 genes representing the majority of the mouse protein-coding transcriptome. We identified 1,191 modulators of prion propagation. While 1,151 modified the expression of both the pathological prion protein, PrPSc , and its cellular counterpart, PrPC , 40 genes selectively affected PrPSc . Of the latter 40 genes, 20 augmented prion production when suppressed. A prominent limiter of prion propagation was the heterogeneous nuclear ribonucleoprotein Hnrnpk. Psammaplysene A (PSA), which binds Hnrnpk, reduced prion levels in cultured cells and protected them from cytotoxicity. PSA also reduced prion levels in infected cerebellar organotypic slices and alleviated locomotor deficits in prion-infected Drosophila melanogaster expressing ovine PrPC . Hence, genome-wide QUIPPER-based perturbations can discover actionable cellular pathways involved in prion propagation. Further, the unexpected identification of a prion-controlling ribonucleoprotein suggests a role for RNA in the generation of infectious prions.

Cancer cells reprogram their metabolism to support growth and invasion. While previous work has highlighted how single altered reactions and pathways can drive tumorigenesis, it remains unclear how individual changes propagate at the network level and eventually determine global metabolic activity. To characterize the metabolic lifestyle of cancer cells across pathways and genotypes, we profiled the intracellular metabolome of 180 pan-cancer cell lines grown in identical conditions. For each cell line, we estimated activity for 49 pathways spanning the entirety of the metabolic network. Upon clustering, we discovered a convergence into only two major metabolic types. These were functionally confirmed by 13 C-flux analysis, lipidomics, and analysis of sensitivity to perturbations. They revealed that the major differences in cancers are associated with lipid, TCA cycle, and carbohydrate metabolism. Thorough integration of these types with multiomics highlighted little association with genetic alterations but a strong association with markers of epithelial-mesenchymal transition. Our analysis indicates that in absence of variations imposed by the microenvironment, cancer cells adopt distinct metabolic programs which serve as vulnerabilities for therapy., Molecular Systems Biology, 18 (11), ISSN:1744-4292

Extending knowledge on ecosystem stability to larger spatial scales is urgently needed because present local-scale studies are generally ineffective in guiding management and conservation decisions of an entire region with diverse plant communities. We investigated stability of plant productivity across spatial scales and hierarchical levels of organization and analyzed impacts of dominant species, species diversity, and climatic factors using a multisite survey of Inner Mongolian grassland. We found that regional stability across distant local communities was related to stability and asynchrony of local communities. Using only dominant instead of all-species dynamics explained regional stability almost equally well. The diversity of all or only dominant species had comparatively weak effects on stability and synchrony, whereas a lower mean and higher variation of precipitation destabilized regional and local communities by reducing population stability and synchronizing species dynamics. We demonstrate that, for semi-arid temperate grassland with highly uneven species abundances, the stability of regional communities is increased by stability and asynchrony of local communities and these are more affected by climate rather than species diversity. Reduced amounts and increased variation of precipitation in the future may compromise the sustainable provision of ecosystem services to human well-being in this region.

The quantification of a species’ trophic niche is important to understand the species ecology and its interactions with the ecosystem it resides in. Despite the high frequency of long-finned pilot whale (Globicephala melas edwardii) strandings on the Aotearoa New Zealand coast, their trophic niche remains poorly understood. To assess the isotopic niche of G. m. edwardii within New Zealand, ontogenetic (sex, total body length, age, maturity status, reproductive group) and spatiotemporal (stranding location, stranding event, and stranding year) variation were investigated. Stable isotopes of carbon (δ13C) and nitrogen (δ15N) were examined from skin samples of 125 G. m. edwardii (67 females and 58 males) collected at mass-stranding events at Onetahua Farewell Spit in 2009 (n = 20), 2011 (n = 20), 2014 (n = 27) and 2017 (n = 20) and at Rakiura Stewart Island in 2010 (n = 19) and 2011 (n = 19). Variations in δ34S values were examined for a subset of 36 individuals. General additive models revealed that stranding event was the strongest predictor for δ13C and δ15N values, whilst sex was the strongest predictor of δ34S isotopic values. Although similar within years, δ13C values were lower in 2014 and 2017 compared to all other years. Furthermore, δ15N values were higher within Farewell Spit 2017 compared to any other stranding event. This suggests that the individuals stranded in Farewell Spit in 2017 may have been feeding at a higher trophic level, or that the nitrogen baseline may have been higher in 2017 than in other years. Spatiotemporal differences explained isotopic variation of G. m. edwardii in New Zealand waters better than ontogenetic factors.

Reptarenaviruses cause boid inclusion body disease (BIBD), a poten- tially fatal disease, occurring in captive constrictor snakes boas and pythons worldwide. Classical BIBD, characterized by the formation of pathognomonic cyto- plasmic inclusion bodies (IBs), occurs mainly in boas, whereas in pythons, for example, reptarenavirus infection most often manifests as central nervous system signs with limited IB formation. The natural hosts of reptarenaviruses are unknown, although free-ranging/wild constrictor snakes are among the suspects. Here, we report BIBD with reptarenavirus infection in indigenous captive and wild boid snakes in Costa Rica using histology, immunohistology, transmission electron microscopy, and next-generation sequencing (NGS). The snakes studied represented diagnostic post- mortem cases of captive and wild-caught snakes since 1989. The results from NGS on archival paraffin blocks confirm that reptarenaviruses were already present in wild boa constrictors in Costa Rica in the 1980s. Continuous sequences that were de novo assembled from the low-quality RNA obtained from paraffin-embedded tissue allowed the identification of a distinct pair of reptarenavirus S and L segments in all studied animals; in most cases, reference assembly could recover almost complete segments. Sampling of three prospective cases in 2018 allowed an examination of fresh blood or tissues and resulted in the identification of additional reptarenavirus segments and hartmanivirus coinfection. Our results show that BIBD is not only a disease of captive snakes but also occurs in indigenous wild constrictor snakes in Costa Rica, suggesting boa constrictors to play a role in natural reptarenavirus circulation. Los reptarenavirus causan la enfermedad del cuerpo de inclusión de las boas (BIBD), una enfermedad potencialmente mortal que se produce en las serpientes constrictoras cautivas, boas y pitones, de las serpientes constrictoras en cautividad, las boas y las pitones, en todo el mundo. La BIBD clásica, caracterizada por la formación de cuerpos de inclusión cito de inclusión patognomónicos, se da principalmente en las boas, mientras que en las pitones, por ejemplo, la infección por reptarenavirus se manifiesta más a menudo como con una formación limitada de IB. Los huéspedes naturales de los reptarenavirus son desconocidos, aunque las serpientes constrictoras salvajes y en libertad están entre los sospechosos. En este caso, informamos de la existencia de de reptarenavirus en serpientes constrictoras autóctonas y silvestres de Costa Rica. Costa Rica utilizando histología, inmunohistología, microscopía electrónica de transmisión y secuenciación de próxima generación (NGS). Las serpientes estudiadas representaban casos de diagnóstico post de diagnóstico de serpientes cautivas y silvestres desde 1989. Los resultados de la NGS en de parafina de archivo confirman que los reptarenavirus ya estaban presentes en las boas en Costa Rica en la década de 1980. Las secuencias continuas que se ensamblaron de novo a partir del ARN de baja calidad obtenido de los tejidos embebidos en parafina, permitieron identificar un par de segmentos S y L del reptarenavirus en todos los animales estudiados. En la mayoría de los casos, el ensamblaje de referencia pudo recuperar segmentos casi completos. El muestreo de tres casos prospectivos en 2018 permitió un examen de sangre o tejidos y dio lugar a la identificación de segmentos adicionales de reptarenavirus y coinfección por hartmanivirus. Nuestros resultados muestran que la BIBD no es solo una enfermedad de las serpientes cautivas sino que también se da en las serpientes constrictoras silvestres de Costa Rica, lo que sugiere que las boas constrictoras desempeñan un papel en la circulación natural de reptarenavirus. Universidad Nacional, Costa Rica Escuela de Medicina Veterinaria

Childhood and adolescence are critical stages of the human lifespan, in which fundamental neural reorganizational processes take place. A substantial body of literature investigated accompanying neurophysiological changes, focusing on the most dominant feature of the human EEG signal: the alpha oscillation. Recent developments in EEG signal-processing show that conventional measures of alpha power are confounded by various factors and need to be decomposed into periodic and aperiodic components, which represent distinct underlying brain mechanisms. It is therefore unclear how each part of the signal changes during brain maturation. Using multivariate Bayesian generalized linear models, we examined aperiodic and periodic parameters of alpha activity in the largest openly available pediatric dataset (N=2529, age 5-22 years) and replicated these findings in a preregistered analysis of an independent validation sample (N=369, age 6-22 years). First, the welldocumented age-related decrease in total alpha power was replicated. However, when controlling for the aperiodic signal component, our findings provided strong evidence for an age-related increase in the aperiodic-adjusted alpha power. As reported in previous studies, also relative alpha power revealed a maturational increase, yet indicating an underestimation of the underlying relationship between periodic alpha power and brain maturation. The aperiodic intercept and slope decreased with increasing age and were highly correlated with total alpha power. Consequently, earlier interpretations on age-related changes of total alpha power need to be reconsidered, as elimination of active synapses rather links to decreases in the aperiodic intercept. Instead, analyses of diffusion tensor imaging data indicate that the maturational increase in aperiodic-adjusted alpha power is related to increased thalamocortical connectivity. Functionally, our results suggest that increased thalamic control of cortical alpha power is linked to improved attentional performance during brain maturation., eLife, 11, ISSN:2050-084X

The nuclear pore complex (NPC) is the central portal for macromolecular exchange between the nucleus and cytoplasm. In all eukaryotes, NPCs assemble into an intact nuclear envelope (NE) during interphase, but the process of NPC biogenesis remains poorly characterized. Furthermore, little is known about how NPC assembly leads to the fusion of the outer and inner NE, and no factors have been identified that could trigger this event. Here, we characterize the transmembrane protein Brl1 as an NPC assembly factor required for NE fusion in budding yeast. Brl1 preferentially associates with NPC assembly intermediates and its depletion halts NPC biogenesis, leading to NE herniations that contain inner and outer ring nucleoporins but lack the cytoplasmic export platform. Furthermore, we identify an essential amphipathic helix in the luminal domain of Brl1 that mediates interactions with lipid bilayers. Mutations in this amphipathic helix lead to NPC assembly defects, and cryo-electron tomography analyses reveal multilayered herniations of the inner nuclear membrane with NPC-like structures at the neck, indicating a failure in NE fusion. Taken together, our results identify a role for Brl1 in NPC assembly and suggest a function of its amphipathic helix in mediating the fusion of the inner and outer nuclear membranes., eLife, 11, ISSN:2050-084X

Inter-individual differences in gut microbiota composition are hypothesized to generate variation in host fitness – a premise for the evolution of host-gut microbe symbioses. However, recent evidence suggests that gut microbial communities are highly dynamic, challenging the notion that individuals harbour unique gut microbial phenotypes. Leveraging a long-term dataset of wild meerkats, we reconcile these concepts by demonstrating that the relative importance of identity for shaping gut microbiota phenotypes depends on the temporal scale. Across meerkat lifespan, year-to-year variation overshadowed the effects of identity and social group in predicting gut microbiota composition, with identity explaining on average less than 2% of variation. However, identity was the strongest predictor of microbial phenotypes over short sampling intervals (< 2 months), predicting on average 20% of variation. The effect of identity was also dependent on meerkat age, with the gut microbiota becoming more individualized and stable as meerkats aged. Nevertheless, whilst the predictive power of identity was negligible after two months, gut microbiota composition remained weakly individualised compared to that of other meerkats for up to one year. These findings illuminate the degree to which individualised gut microbial signatures can be expected, with important implications for the time frames over which gut microbial phenotypes may mediate host physiology, behaviour and fitness in natural populations., acceptedVersion

The tumour microenvironment and genetic alterations collectively influence drug efficacy in cancer, but current evidence is limited and systematic analyses are lacking. Using chronic lymphocytic leukaemia (CLL) as a model disease, we investigated the influence of 17 microenvironmental stimuli on 12 drugs in 192 genetically characterised patient samples. Based on microenvironmental response, we identified four subgroups with distinct clinical outcomes beyond known prognostic markers. Response to multiple microenvironmental stimuli was amplified in trisomy 12 samples. Trisomy 12 was associated with a distinct epigenetic signature. Bromodomain inhibition reversed this epigenetic profile and could be used to target microenvironmental signalling in trisomy 12 CLL. We quantified the impact of microenvironmental stimuli on drug response and their dependence on genetic alterations, identifying interleukin 4 (IL4) and Toll-like receptor (TLR) stimulation as the strongest actuators of drug resistance. IL4 and TLR signalling activity was increased in CLL-infiltrated lymph nodes compared with healthy samples. High IL4 activity correlated with faster disease progression. The publicly available dataset can facilitate the investigation of cell-extrinsic mechanisms of drug resistance and disease progression.

While in Switzerland, pneumococcal vaccination is recommended for adult patients with axial spondyloarthritis (axSpA) treated with biological drugs, since 2014, little is known about the vaccination status of this specific patient population. This study assessed their vaccination status as part of a larger online survey by the ankylosing spondylitis association of Switzerland (SVMB). Out of 1560 who participated in the survey, 834 (53.5%) were eligible for the analysis. Vaccine coverage was low at 32.5% (271/834). Women and patients who got a flu shot every year were more likely to be covered. Age was negatively associated with being vaccinated. Most (54.2%; 147/271) were vaccinated by their general practitioner. Almost two-thirds of those who had not received the vaccine stated that it had not been offered to them (64.1%; 302/471). In summary, the vaccination coverage is low, but might be increased if the vaccine was offered systematically by general practitioners and specialists.

Mammarenaviruses cause a persistent infection in their natural rodent and bat hosts. Reptarenaviruses cause boid inclusion body disease (BIBD) in constrictor snakes, but it is unclear whether snakes are the natural host of these viruses. Mammarenaviruses establish a persistent infection in their rodent and bat hosts, and the evidence suggests that reptarenaviruses and hartmaniviruses found in captive snakes act similarly. In snakes, reptarenaviruses cause boid inclusion body disease (BIBD), which is often associated with secondary infections. Snakes with BIBD usually carry more than a single pair of reptarenavirus S and L segments and occasionally demonstrate hartmanivirus coinfection. Here, we reported the generation of cell lines persistently infected with a single or two reptarenavirus(es) and a cell line with persistent reptarenavirus-hartmanivirus coinfection. By RT-PCR we demonstrated that the amount of viral RNA within the persistently infected cells remains at levels similar to those observed following initial infection. Using antibodies against the glycoproteins (GPs) and nucleoprotein (NP) of reptarenaviruses, we studied the levels of viral protein in cells passaged 10 times after the original inoculation and observed that the expression of GPs declines dramatically during persistent infection, unlike the expression of NP. Immunofluorescence (IF) staining served to demonstrate differences in the distribution of NP within the persistently infected compared to freshly infected cells. IF staining of cells inoculated with the viruses secreted from the persistently infected cell lines produced similar NP staining compared to cells infected with a traditionally passaged virus, suggesting that the altered NP expression pattern of persistently infected cells does not relate to changes in the virus. The cell cultures described herein can serve as tools for studying the coinfection and superinfection interplay between reptarenaviruses and studying the BIBD pathogenesis mechanisms. IMPORTANCE Mammarenaviruses cause a persistent infection in their natural rodent and bat hosts. Reptarenaviruses cause boid inclusion body disease (BIBD) in constrictor snakes, but it is unclear whether snakes are the natural host of these viruses. In this study, we showed that reptarenaviruses established a persistent infection in cultured Boa constrictor cells and that the persistently infected cells continued to produce infectious virus. Our results showed that persistent infection results from subsequent passaging of cells inoculated with a single reptarenavirus, two reptarenaviruses, or even when inoculating the cells with reptarenavirus and hartmanivirus (another arenavirus genus). The results further suggested that coinfection would not result in overt competition between the different reptarenaviruses, thus helping to explain the frequent reptarenavirus coinfections in snakes with BIBD. The established cell culture models of persistent infection could help to elucidate the role of coinfection and superinfection and potential immunosuppression as the pathogenic mechanisms behind BIBD.

Shark–cetacean trophic interactions, preserved as bite marks in the fossil record, mostly correspond to isolated or fragmentary findings that bear limited information about major trophic patterns or roles. Here, we provide evidence of focalized foraging by sharks in the form of tooth bite marks over physeteroids fossil bones from the late Miocene of Peru. These findings indicate that sharks were targeting the forehead of coeval physeteroids to actively feed on their lipid-rich nasal complexes. Miocene physeteroids displayed a broad diversity, including giant predatorial forms, small benthic foragers and suction feeders. Like their extant relatives, these animals exhibited enlarged fatty forehead organs responsible for their sound production capabilities, thus evolving taxon-specific cranial architecture. Bite marks are found on the cranial bones where these structures were attached, indicating that sharks actively targeted this region; but also, in areas that would only be accessible following the consumption of the surrounding soft tissues. The shape of the bite marks and their distribution suggests a series of consecutive scavenging events by individuals of different shark species. Similar bite patterns can be recognized on other Miocene physeteroids fossils from across the globe, suggesting that sharks actively exploited physeteroid carcasses as fat sources.

Species occurring in sympatry and relying on similar and limited resources may partition resource use to avoid overlap and interspecific competition. Aotearoa, New Zealand hosts an extraordinarily rich marine megafauna, including 50% of the world’s cetacean species. In this study, we used carbon and nitrogen stable isotopes as ecological tracers to investigate isotopic niche overlap between 21 odontocete (toothed whale) species inhabiting neritic, mesopelagic, and bathypelagic waters. Results showed a clear niche separation for the bathypelagic Gray’s beaked whales (Mesoplodon grayi) and sperm whales (Physeter macrocephalus), but high isotopic niche overlap and potential interspecific competition for neritic and mesopelagic species. For these species, competition could be reduced via temporal or finer-scale spatial segregation or differences in foraging behaviour. This study represents the first insights into the coexistence of odontocetes in a biodiverse hotspot. The data presented here provide a critical baseline to a system already ongoing ecosystem change via ocean warming and subsequent effects on prey abundance and distributions.

The ability to quantify partial pressure of oxygen (pO

3’ end formation of most eukaryotic mRNAs is dependent on the assembly of a ∼1.5 megadalton multiprotein complex, that catalyzes the coupled reaction of pre-mRNA cleavage and polyadenylation. In mammals, the cleavage and polyadenylation specificity factor (CPSF) constitutes the core of the 3’ end processing machinery onto which the remaining factors, including cleavage stimulation factor (CstF) and poly(A) polymerase (PAP), assemble. These interactions are mediated by Fip1, a CPSF subunit characterized by high degree of intrinsic disorder. Here, we report two crystal structures revealing the interactions of human Fip1 (hFip1) with CPSF30 and CstF77. We demonstrate that CPSF contains two copies of hFip1, each binding to the zinc finger (ZF) domains 4 and 5 of CPSF30. Using polyadenylation assays we show that the two hFip1 copies are functionally redundant in recruiting one copy of PAP, thereby increasing the processivity of RNA polyadenylation. We further show that the interaction between hFip1 and CstF77 is mediated via a short motif in the N-terminal “acidic” region of hFip1. In turn, CstF77 competitively inhibits CPSF-dependent PAP recruitment and 3’ polyadenylation. Taken together, these results provide a structural basis for the multivalent scaffolding and regulatory functions of hFip1 in 3’ end processing.

Deconstruction of plant cell walls is imperative to global carbon cycling and sustainability efforts. Selected microbes degrade plant fibers using extremely efficient multi-enzymatic cellulosomes assemblies. Organization of cellulosomes on the bacterial cell surface and their ecological regulation remain elusive. By combining structural methodologies with molecular and biochemical approaches on the canonical Clostridium thermocellum system, we provide an unprecedented view into the in-situ structure and distribution of cellulosomal enzymes while interacting with their cellulosic substrate during fiber degradation. Structural exploration of growing cultures revealed isogenic phenotypic heterogeneity of cellulosome organization on single cells across the bacterial population, suggesting a division-of labor strategy driven by product-dependent dynamics. This study demonstrates how structural biology under near-physiological conditions can be employed to develop ecological hypotheses to understand microbial plant-fiber degradation at the single-cell nanoscale level.One Sentence SummaryThis study contributes critical insights into the in-situ organization of cellulosomes and their cellulosic substrates and provides evidence for phenotypic heterogeneity, with dynamic, growth phase-dependent organization of the fiber-degrading machinery.

From the management of microtubules to the production of pathological species: liquid-liquid phase separation may tune the behavior of the protein tau in health and neurodegenerative disease. In this issue of The EMBO Journal, Hochmair et al (2022) demystify important aspects of tau condensate compilation.

Filifactor alocis is a Gram-positive asaccharolytic, obligate anaerobic rod that has been isolated from a variety of oral infections including periodontitis, peri-implantitis, and odontogenic abscesses. As a newly emerging pathogen, its type strain has been investigated for pathogenic properties, yet little is known about its virulence variations among strains. We previously screened the whole genome of nine clinical oral isolates and a reference strain of F. alocis, and they expressed a novel RTX toxin, FtxA. In the present study, we aimed to use label-free quantification proteomics to characterize the full proteome of those ten F. alocis strains. A total of 872 proteins were quantified, and 97 among them were differentially expressed in FtxA-positive strains compared with the negative strains. In addition, 44 of these differentially expressed proteins formed 66 pairs of associations based on their predicted functions, which included clusters of proteins with DNA repair/mediated transformation and catalytic activity-related function, indicating different biosynthetic activities among strains. FtxA displayed specific interactions with another six intracellular proteins, forming a functional cluster that could discriminate between FtxA-producing and non-producing strains. Among them were FtxB and FtxD, predicted to be encoded by the same operon as FtxA. While revealing the broader qualitative and quantitative proteomic landscape of F. alocis, this study also sheds light on the deeper functional inter-relationships of FtxA, thus placing this RTX family member into context as a major virulence factor of this species., Pathogens, 11 (5), ISSN:2076-0817

Calcium ions function as a key second messenger ion in eukaryotes. Spatially and temporally defined cytoplasmic Ca2+ signals are shaped through the concerted activity of ion channels, exchangers, and pumps in response to diverse stimuli; these signals are then decoded through the activity of Ca2+-binding sensor proteins. In plants, Ca2+ signaling is central to both pattern- and effector-triggered immunity, with the generation of characteristic cytoplasmic Ca2+ elevations in response to potential pathogens being common to both. However, despite their importance, and a long history of scientific interest, the transport proteins that shape Ca2+ signals and their integration remain poorly characterized. Here, we discuss recent work that has both shed light on and deepened the mysteries of Ca2+ signaling in plant immunity.

The Chlamydiae are a phylum of obligate intracellular, Gram-negative bacteria with a biphasic lifecycle [...]

Borna disease (BD), a frequently fatal neurologic disorder caused by Borna disease virus 1 (BoDV-1), has been observed for decades in horses, sheep, and other mammals in certain regions of Europe. The bicoloured white-toothed shrew (Crocidura leucodon) was identified as a persistently infected species involved in virus transmission. Recently, BoDV-1 attracted attention as a cause of fatal encephalitis in humans. Here we report investigations on BoDV-1 infected llamas from a farm in a BD endemic area of Switzerland, and alpacas from holdings in a region of Germany where BD was last seen in the 1960s but not thereafter. All New World camelids showed apathy and abnormal behaviour, necessitating euthanasia. Histologically, severe non-suppurative meningoencephalitis with neuronal Joest-Degen inclusion bodies was observed. BoDV-1 was confirmed by immunohistology, RT-qPCR, and sequencing in selected animals. Analysis of the llama herd over 20 years showed that lossesdue to clinically suspected BD increased within the last decade. BoDV-1 whole-genome sequences from one Swiss llama and one German alpaca, and – for comparison – from one Swiss horse and one German shrew were established. They represent the first published whole-genome sequences of BoDV-1 clusters 1B and 3, respectively. Our analysis suggests that New World camelids may have a role as a sentinel species for BoDV-1 infection, even when symptomatic cases are lacking in other animal species.

Summary Pluripotent stem cell-derived human cortical organoids allow for the analysis of stem cell behavior and neurogenesis in neural tissues. Delivery of plasmid DNA into organoids permits visualization of individual cells, characterization of cellular components, and manipulation of gene expression. We describe a protocol to transfect cells inside organoids with plasmid DNA using micro-injection and electroporation, allowing for DNA delivery to cells within cortical units. This protocol was optimized for cortical organoids; however, it may be adapted to other organoid models. For complete details on the use and execution of this protocol, please refer to Denoth-Lippuner et al. (2021), Graphical abstract, Highlights • Plasmid injection of pluripotent stem cell-derived human cortical organoids • Electroporation of human cortical organoids • Genetic manipulation of progenitors and their progeny in cortical organoids • Protocol enables robust DNA transfection of human cortical organoids, Pluripotent stem cell-derived human cortical organoids allow for the analysis of stem cell behavior and neurogenesis in neural tissues. Delivery of plasmid DNA into organoids permits visualization of individual cells, characterization of cellular components, and manipulation of gene expression. We describe a protocol to transfect cells inside organoids with plasmid DNA using micro-injection and electroporation, allowing for DNA delivery to cells within cortical units. This protocol was optimized for cortical organoids; however, it may be adapted to other organoid models.

Due to the low stoichiometry and highly transient nature of protein phosphorylation it is challenging to capture the dynamics and complexity of phosphorylation events on a systems level. Here, we present an optimized protocol to measure virus-induced phosphorylation events with high sensitivity using label free quantification-based phosphoproteomics. Specifically, we describe filter assisted protein digestion (FASP), enrichment of phosphopeptides, mass spectrometry, and subsequent bioinformatic analysis. For complete details on the use and execution of this protocol, please refer to Hunziker et al. (2022)., STAR Protocols, 3 (1), ISSN:2666-1667

Hepatitis E virus (HEV) is the causative agent of an acute and in most cases self-limiting hepatitis. Of the four major HEV genotypes that infect humans, genotype 3 and 4 are zoonotic and have been identified in humans but predominantly in pigs and wild boar, which are considered the main reservoirs. However, the known host range of zoonotic HEV may be increasing to comprise additional species, including companion animals. Several studies have identified contact with dogs as a risk factor for HEV infection in humans, yet information on the occurrence of HEV in Swiss dogs is lacking. To examine a possible risk of exposure, this study was designed to assess the seroprevalence of HEV in 84 Swiss dogs. Serum and plasma samples collected from four veterinary clinics were screened for HEV-specific antibodies by HEV-antibody ELISA test kit. In addition, information of 22 dogs regarding the country of origin, the type of dog feed and any history of hunting was recorded. Samples from seropositive animals were also screened for the presence of HEV RNA by quantitative real-time RT-PCR (qRT-PCR). Overall, 38% (32 of 84) of the dogs tested seropositive for anti-HEV, indicating exposure to HEV. Among the 22 dogs for which information was available, HEV-specific antibodies were detected in three of five dogs that were born abroad, in one of two dogs that were fed a raw meat-based diet, and in one hunting dog. No viral RNA could be detected in any of the serum and plasma samples; thus, the genotype of the strains remained undetermined. This study provides further evidence for canine exposure and susceptibility to HEV and highlights the need to further assess the risks of HEV transmission to humans with contact to dogs.

The incidence of chronic kidney disease (CKD) is associated with major abnormalities in circulating lipoproteins and renal lipid metabolism. This article elaborates on the mechanisms of CKD and lipid uptake abnormalities. The viewpoint we supported is that lipid abnormalities directly cause CKD, resulting in forming a vicious cycle. On the theoretical and experiment fronts, this inference has been verified by elaborately elucidating the role of lipid intake and accumulation as well as their influences on CKD. Taken together, these findings suggest that further understanding of lipid metabolism in CKD may lead to novel therapeutic approaches.

In competitive alpine skiing, a superior antirotation and rear-chain stabilization capacity is essential to constantly remain in dynamic equilibrium while skiing and to counteract the ski-specific adverse loading patterns of the back. As such, skiers’ trunk stabilization performance during deadbug bridging (DBB) exercises has been shown to be associated with both skiing performance and overuse complaints of the lower back in skiers under 16 years of age (U16). However, to date, little is known about the corresponding stabilization abilities in younger skiers, i.e., 6- to 15-year-old skiers. As part of a biomechanical field experiment during a national off-snow fitness competition, a total of 101 youth competitive alpine skiers were tested with respect to their trunk stabilization performance during DDB exercise. The maximum contralateral displacement of the pelvic drop during leg lift (DBBdisplacement) was quantified using reflective markers and a motion capture system (Vicon, Oxford, UK). Potential age group and sex differences in DBBdisplacement were assessed using analysis of variance (ANOVA) at p < 0.05. Within each subgroup, the associations of DBBdisplacement with age, anthropometrics and maturity offset were analysed using Pearson’s correlation (p < 0.05). Female skiers under 15 years of age (U15) showed better DBB performance than male U15 skiers, while there was no sex difference at the under 10-year (U10) level. In female U10 skiers, DBBdisplacement was moderately associated with body height, while in all other subgroups, no confounding associations with anthropometrics or biological maturation were found. Biomechanically quantifying DBB performance may be considered a feasible and nonconfounded screening test approach in young skiers older than 6 years. Body height may represent a confounding bias in exclusively the U10 female skier cohort and, therefore, should be considered when interpreting the test results. In summary, this study provided sport-specific normative reference data that may be of equal interest to both researchers and sport practitioners.

Cells encountering stressful situations activate the integrated stress response (ISR) pathway to limit protein synthesis and redirect translation to better cope. The ISR has also been implicated in cancers, but redundancies in the stress-sensing kinases that trigger the ISR have posed hurdles to dissecting physiological relevance. To overcome this challenge, we targeted the regulatory node of these kinases, namely, the S51 phosphorylation site of eukaryotic translation initiation factor eIF2α and genetically replaced eIF2α with eIF2α-S51A in mouse squamous cell carcinoma (SCC) stem cells of skin. While inconsequential under normal growth conditions, the vulnerability of this ISR-null state was unveiled when SCC stem cells experienced proteotoxic stress. Seeking mechanistic insights into the protective roles of the ISR, we combined ribosome profiling and functional approaches to identify and probe the functional importance of translational differences between ISR-competent and ISR-null SCC stem cells when exposed to proteotoxic stress. In doing so, we learned that the ISR redirects translation to centrosomal proteins that orchestrate the microtubule dynamics needed to efficiently concentrate unfolded proteins at the microtubule-organizing center so that they can be cleared by the perinuclear degradation machinery. Thus, rather than merely maintaining survival during proteotoxic stress, the ISR also functions in promoting cellular recovery once the stress has subsided. Remarkably, this molecular program is unique to transformed skin stem cells, hence exposing a vulnerability in cancer that could be exploited therapeutically.

Cell therapy has long been an emerging treatment paradigm in experimental neurobiology. However, cell transplantation studies often rely on end-point measurements and can therefore only evaluate longitudinal changes of cell migration and survival to a limited extent. This paper provides a reliable, minimally invasive protocol to transplant and longitudinally track neural progenitor cells (NPCs) in the adult mouse brain. Before transplantation, cells are transduced with a lentiviral vector comprising a bioluminescent (firefly-luciferase) and fluorescent (green fluorescent protein [GFP]) reporter. The NPCs are transplanted into the right cortical hemisphere using stereotaxic injections in the sensorimotor cortex. Following transplantation, grafted cells were detected through the intact skull for up to five weeks (at days 0, 3, 14, 21, 35) with a resolution limit of 6,000 cells using in vivo bioluminescence imaging. Subsequently, the transplanted cells are identified in histological brain sections and further characterized with immunofluorescence. Thus, this protocol provides a valuable tool to transplant, track, quantify, and characterize cells in the mouse brain., Journal of Visualized Experiments. JoVE (179), ISSN:1940-087X

Objectives: Since the change in the millennium, an increase in cases of alveolar echinococcosis (AE) has been observed in endemic European countries. Previous studies indicate that a significant proportion of the new AE cases have an immunosuppression-associated condition (IAC). The aim of the current study was to determine how IACs impact the number of new AE diagnoses per year and the characteristics of AE at diagnosis and its clinical course at our center. Methods: Retrospective analysis of 189 patients with AE diagnosed between 2000 and 2021 and participating in the Zurich Echinococcosis Cohort Study (ZECS) included clinical characteristics of AE at diagnosis and report of an IAC, as well as the clinical course during follow-up. Results: Of 189 patients participating in this study, 38 had an IAC reported at, or shortly after, AE diagnosis. Over time, there was a steeper increase in the number of newly diagnosed AE patients without an IAC than the number of patients with IAC. Patients with an IAC were older at diagnosis, more frequently had an incidental finding of AE, smaller mean lesion size, and negative Em18 serology. All but two showed favorable outcomes on the last follow-up. Conclusion: IACs have little impact on the increase in new AE cases, as well as on the extent of the disease at diagnosis and clinical course.

Although monitoring of vaccination program performance is usually evaluated by measurement of vaccine coverage, timely uptake is rarely part of this assessment. This study aims to examine the timeliness of the administration of a measles-containing-vaccine (MCV) for 2-year-old children between 2005 and 2019.We used data from the Swiss National Vaccination Coverage Survey 2005-2019 for the study. We defined timely vaccinated as a vaccination administered within the recommended age specified in the Swiss National Vaccination Schedule, with an added tolerance period of 30.4 days for both MCV 1 and 2 doses. The median delay time was estimated by Kaplan-Meier survival curve and examined using log-rank test. A Cox hazard ratio was used to identify factors associated with delay.81% (95% CI:79-82%) of toddlers were timely vaccinated for MCV1 and 82% (95% CI:81-83%) for MCV2 in survey period 2017-2019. Between 2005 and 2019, the median age of vaccinated children ranged between 12.2 and 12.5 and 18.3-22.0 months for MCV1 and MCV2 with median delay of 44 and 38 days, respectively, at the national level. Children in the French-, Italian- and German-speaking regions were vaccinated earlier between 2005 and 2019 for MCV1 (vaccination coverage range before 10 months of age: 1.7-45.9%, 1.2-35.3% and 1.4-15.0%, respectively). Nationality, linguistic regions, and survey periods were the strongest predictive factors related to prolonged delay time.Overall adherence to recommendations has improved over time, as MCV coverage has significantly increased over the years with differences across linguistic regions. Vaccinations were administered earlier and with shorter delay time.

Influenza A virus (IAV) coopts numerous host factors for efficient replication. The cysteine protease cathepsin W (CTSW) has been identified as one host factor required for IAV entry, specifically for the escape of IAVs from late endosomes. However, the substrate specificity of CTSW and the proviral mechanism are thus far unknown. Here, we show that intracellular but not secreted CTSW promotes viral entry. We reveal 79 potential direct and 31 potential indirect cellular target proteins of CTSW using the high-throughput proteomic approach terminal amine isotopic labeling of substrates (TAILS) and determine the cleavage motif shared by the substrates of CTSW. Subsequent integration with data from RNA interference (RNAi) screens for IAV host factors uncovers first insights into the proviral function of CTSW. Notably, CTSW-deficient mice display a 25% increase in survival and a delay in mortality compared to wild-type mice upon IAV infection. Altogether, these findings support the development of drugs targeting CTSW as novel host-directed antiviral therapies. IMPORTANCE Influenza viruses are respiratory pathogens and pose a constant threat to human health. Although antiviral drugs are available for influenza, the emergence and spread of drug-resistant viruses is cause for concern. Therefore, the development of new antivirals with lower chances of their target viruses acquiring resistance is urgently needed to reduce the high morbidity and mortality caused by influenza. Promising alternatives to drugs targeting viral proteins are those directed against host factors required for viral replication. The cysteine protease cathepsin W (CTSW) is an important host factor for IAV replication, and its proteolytic activity is required for fusion of viral and endosomal membranes. In this work, we identify a number of hitherto unknown CTSW substrates, providing new insights into virus-host interactions, and reveal that CTSW might also play a proviral role in an in vivo model. These results support the development of CTSW as a drug target for next-generation antivirals against influenza., Microbiology Spectrum, 10 (4), ISSN:2165-0497

From observations in rodents, it has been suggested that the cellular basis of learning-dependent changes, detected using structural MRI, may be increased dendritic spine density, alterations in astrocyte volume, and adaptations within intracortical myelin. Myelin plasticity is crucial for neurological function, and active myelination is required for learning and memory. However, the dynamics of myelin plasticity and how it relates to morphometric-based measurements of structural plasticity remains unknown. We used a motor skill learning paradigm in male mice to evaluate experience-dependent brain plasticity by voxel-based morphometry (VBM) in longitudinal MRI, combined with a cross-sectional immunohistochemical investigation. Whole-brain VBM revealed nonlinear decreases in gray matter volume (GMV) juxtaposed to nonlinear increases in white matter volume (WMV) within GM that were best modeled by an asymptotic time course. Using an atlas-based cortical mask, we found nonlinear changes with learning in primary and secondary motor areas and in somatosensory cortex. Analysis of cross-sectional myelin immunoreactivity in forelimb somatosensory cortex confirmed an increase in myelin immunoreactivity followed by a return towards baseline levels. Further investigations using quantitative confocal microscopy confirmed these changes specifically to the length density of myelinated axons. The absence of significant histological changes in cortical thickness suggests that nonlinear morphometric changes are likely due to changes in intracortical myelin for which morphometric WMV in somatosensory cortex significantly correlated with myelin immunoreactivity. Together, these observations indicate a nonlinear increase of intracortical myelin during learning and support the hypothesis that myelin is a component of structural changes observed by VBM during learning.

This study aimed to understand parental discourse about vaccination, and to provide guidance for communication that addresses the needs of parents. We analyzed parental discourse on child vaccination in general and tick-borne encephalitis (TBE) specifically in a Swiss parental online community. For this purpose, a data set containing 105k posts written by parents between 2007 and 2019 was analyzed using a combination of linguistic discourse analysis and qualitative content analysis. Results show that parents enter into a multidimensional decision-making process, characterized by elaborate practices of negotiation, consideration of vaccination recommendations as well as six distinct influencing thematic factors (vaccination safety, development and control, effectiveness, epidemiology, necessity, alternatives or additional prevention methods). The study shows a clear pattern of seasonality, with parents talking about TBE vaccination mostly triggered by events such as tick bites in spring and summer. From a public health perspective, the study emphasizes the need for sufficient, balanced, and tailored information about TBE vaccination. Online forums provide valuable information about what matters to parents and when, which can help public health authorities and practitioners provide information according to these concerns and enhance health literacy among parents.