Potent immunogenicity and protective efficacy of a multi-pathogen vaccination targeting Ebola, Sudan, Marburg and Lassa viruse.

Viral haemorrhagic fevers (VHF) pose a significant threat to human health. In recent years, VHF outbreaks caused by Ebola, Marburg and Lassa viruses have caused substantial morbidity and mortality in West and Central Africa. In 2022, an Ebola disease outbreak in Uganda caused by Sudan virus resulted in 164 cases with 55 deaths. In 2023, a Marburg disease outbreak was confirmed in Equatorial Guinea and Tanzania resulting in over 49 confirmed or suspected cases; 41 of which were fatal. There are no clearly defined correlates of protection against these VHF, impeding targeted vaccine development. Any vaccine developed should therefore induce strong and preferably long-lasting humoral and cellular immunity against these viruses. Ideally this immunity should also cross-protect against viral variants, which are known to circulate in animal reservoirs and cause human disease. We have utilized two viral vectored vaccine platforms, an adenovirus (ChAdOx1) and Modified Vaccinia Ankara (MVA), to develop a multi-pathogen vaccine regime against three filoviruses (Ebola virus, Sudan virus, Marburg virus) and an arenavirus (Lassa virus). These platform technologies have consistently demonstrated the capability to induce robust cellular and humoral antigen-specific immunity in humans, most recently in the rollout of the licensed ChAdOx1-nCoV19/AZD1222. Here, we show that our multi-pathogen vaccines elicit strong cellular and humoral immunity, induce a diverse range of chemokines and cytokines, and most importantly, confers protection after lethal Ebola virus, Sudan virus and Marburg virus challenges in a small animal model.

Small molecule induced STING degradation facilitated by the HECT ligase HERC4.

Stimulator of interferon genes (STING) is a central component of the cytosolic nucleic acids sensing pathway and as such master regulator of the type I interferon response. Due to its critical role in physiology and its' involvement in a variety of diseases, STING has been a focus for drug discovery. Targeted protein degradation (TPD) has emerged as a promising pharmacology for targeting previously considered undruggable proteins by hijacking the cellular ubiquitin proteasome system (UPS) with small molecules. Here, we identify AK59 as a STING degrader leveraging HERC4, a HECT-domain E3 ligase. Additionally, our data reveals that AK59 is effective on the common pathological STING mutations, suggesting a potential clinical application of this mechanism. Thus, these findings introduce HERC4 to the fields of TPD and of compound-induced degradation of STING, suggesting potential therapeutic applications.

Drug-induced eRF1 degradation promotes readthrough and reveals a new branch of ribosome quality control.

Suppression of premature termination codons (PTCs) by translational readthrough is a promising strategy to treat a wide variety of severe genetic diseases caused by nonsense mutations. Here, we present two potent readthrough promoters-NVS1.1 and NVS2.1-that restore substantial levels of functional full-length CFTR and IDUA proteins in disease models for cystic fibrosis and Hurler syndrome, respectively. In contrast to other readthrough promoters that affect stop codon decoding, the NVS compounds stimulate PTC suppression by triggering rapid proteasomal degradation of the translation termination factor eRF1. Our results show that this occurs by trapping eRF1 in the terminating ribosome, causing ribosome stalls and subsequent ribosome collisions, and activating a branch of the ribosome-associated quality control network, which involves the translational stress sensor GCN1 and the catalytic activity of the E3 ubiquitin ligases RNF14 and RNF25.

Convergence of multiple RNA-silencing pathways on GW182/TNRC6.

The RNA-binding protein TRIM71/LIN-41 is a phylogenetically conserved developmental regulator that functions in mammalian stem cell reprogramming, brain development, and cancer. TRIM71 recognizes target mRNAs through hairpin motifs and silences them through molecular mechanisms that await identification. Here, we uncover that TRIM71 represses its targets through RNA-supported interaction with TNRC6/GW182, a core component of the miRNA-induced silencing complex (miRISC). We demonstrate that AGO2, TRIM71, and UPF1 each recruit TNRC6 to specific sets of transcripts to silence them. As cellular TNRC6 levels are limiting, competition occurs among the silencing pathways, such that the loss of AGO proteins or of AGO binding to TNRC6 enhances the activities of the other pathways. We conclude that a miRNA-like silencing activity is shared among different mRNA silencing pathways and that the use of TNRC6 as a central hub provides a means to integrate their activities.

Comparative evaluation of clinical and cerebrospinal fluid biomarker characteristics in rapidly and non-rapidly progressive Alzheimer's disease.

BACKGROUND: Rapidly progressive forms of Alzheimer's disease (rpAD) are increasingly recognized and may have a prevalence of up to 30% of patients among all patients with Alzheimer's disease (AD). However, insights about risk factors, underlying pathophysiological processes, and clinical characteristics of rpAD remain controversial. This study aimed to gain a comprehensive picture of rpAD and new insights into the clinical manifestation to enable a better interpretation of disease courses in clinical practice as well as in future clinical studies. METHODS: Patients (n = 228) from a prospective observational study on AD were selected and categorized into rpAD (n = 67) and non-rpAD (n = 161) disease groups. Patients were recruited through the German Creutzfeldt-Jakob disease surveillance center and the memory outpatient clinic of the Göttingen University Medical Center, representing diverse phenotypes of the AD population. Biomarkers and clinical presentation were assessed using standardized protocols. A drop of ≥ MMSE 6 points within 12 months defined rapid progressors. RESULTS: Lower CSF Amyloid beta 1-42 concentrations (p = 0.048), lower Amyloid beta 42/40 ratio (p = 0.038), and higher Tau/Amyloid-beta 1-42 ratio, as well as pTau/Amyloid-beta 1-42 ratio (each p = 0.004) were associated with rpAD. Analyzes in a subset of the cohort (rpAD: n = 12; non-rpAD: n = 31) showed higher CSF NfL levels in rpAD (p = 0.024). Clinically, rpAD showed earlier impairment of functional abilities (p < 0.001) and higher scores on the Unified Parkinson's Disease Rating Scale III (p < 0.001), indicating pronounced extrapyramidal motor symptoms. Furthermore, cognitive profiles (adjusted for overall cognitive performance) indicated marked deficits in semantic (p = 0.008) and phonematic (0.023) verbal fluency tests as well as word list learning (p = 0.007) in rpAD compared to non-rpAD. The distribution of APOE genotypes did not differ significantly between groups. CONCLUSIONS: Our results suggest that rpAD is associated with distinct cognitive profiles, earlier occurrence of non-cognitive symptoms, extrapyramidal motoric disturbance, and lower Amyloid-beta 1-42 concentrations in the CSF. The findings may help to characterize a distinct phenotype of rpAD and estimate prognosis based on clinical characteristics and biomarker results. However, an important future goal should be a unified definition for rpAD to enable targeted study designs and better comparability of the results.

Identifying hotspots for ecosystem restoration across heterogeneous tropical savannah-dominated regions.

There is high potential for ecosystem restoration across tropical savannah-dominated regions, but the benefits that could be gained from this restoration are rarely assessed. This study focuses on the Brazilian Cerrado, a highly species-rich savannah-dominated region, as an exemplar to review potential restoration benefits using three metrics: net biomass gains, plant species richness and ability to connect restored and native vegetation. Localized estimates of the most appropriate restoration vegetation type (grassland, savannah, woodland/forest) for pasturelands are produced. Carbon sequestration potential is significant for savannah and woodland/forest restoration in the seasonally dry tropics (net biomass gains of 58.2 ± 37.7 and 130.0 ± 69.4 Mg ha-1). Modelled restoration species richness gains were highest in the central and south-east of the Cerrado for savannahs and grasslands, and in the west and north-west for woodlands/forests. The potential to initiate restoration projects across the whole of the Cerrado is high and four hotspot areas are identified. We demonstrate that landscape restoration across all vegetation types within heterogeneous tropical savannah-dominated regions can maximize biodiversity and carbon gains. However, conservation of existing vegetation is essential to minimizing the cost and improving the chances of restoration success. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.

Mapping native and non-native vegetation in the Brazilian Cerrado using freely available satellite products.

Native vegetation across the Brazilian Cerrado is highly heterogeneous and biodiverse and provides important ecosystem services, including carbon and water balance regulation, however, land-use changes have been extensive. Conservation and restoration of native vegetation is essential and could be facilitated by detailed landcover maps. Here, across a large case study region in Goiás State, Brazil (1.1 Mha), we produced physiognomy level maps of native vegetation (n = 8) and other landcover types (n = 5). Seven different classification schemes using different combinations of input satellite imagery were used, with a Random Forest classifier and 2-stage approach implemented within Google Earth Engine. Overall classification accuracies ranged from 88.6-92.6% for native and non-native vegetation at the formation level (stage-1), and 70.7-77.9% for native vegetation at the physiognomy level (stage-2), across the seven different classifications schemes. The differences in classification accuracy resulting from varying the input imagery combination and quality control procedures used were small. However, a combination of seasonal Sentinel-1 (C-band synthetic aperture radar) and Sentinel-2 (surface reflectance) imagery resulted in the most accurate classification at a spatial resolution of 20 m. Classification accuracies when using Landsat-8 imagery were marginally lower, but still reasonable. Quality control procedures that account for vegetation burning when selecting vegetation reference data may also improve classification accuracy for some native vegetation types. Detailed landcover maps, produced using freely available satellite imagery and upscalable techniques, will be important tools for understanding vegetation functioning at the landscape scale and for implementing restoration projects.

Indigenous and local communities can boost seed supply in the UN decade on ecosystem restoration.

The UN Decade of Ecosystem Restoration is poised to trigger the recovery of ecosystem services and transform structural injustices across the world in a way unparalleled in human history. The inclusion of diverse Indigenous and local communities to co-create robust native seed supply systems is the backbone to achieve the goals for the Decade. Here we show how community-based organizations have co-developed native seed supply strategies for landscape restoration from the bottom-up. We draw on the interconnections over two decades of seed networks in Brazil and the emerging Indigenous participation in native seed production in Australia. From an environmental justice perspective, we provide a participatory seed supply approach for local engagement, noting local geographical, social and cultural contexts. Meeting large-scale restoration goals requires the connection between local seed production and collaborative platforms to negotiate roles, rights and responsibilities between stakeholders. An enduring native seed supply must include a diversity of voices and autonomy of community groups that builds equitable participation in social, economic, and environmental benefits.

Putting fire on the map of Brazilian savanna ecoregions.

The Brazilian savanna (Cerrado) is considered the most floristically diverse savanna in the world, home to more than seven thousand species. The region is a mosaic of savannas, grasslands and forests whose unique biophysical and landscape attributes are on the basis of a recent ecoregional map, paving the way to improved region-based strategies for land management actions. However, as a fire-prone ecosystem, Cerrado owes much of its distribution and ecological properties to the fire regime and contributes to an important parcel of South America burned area. Accordingly, any attempt to use ecoregion geography as a guide for management strategies should take fire into account, as an essential variable. The main aim of this study is to complement the ecoregional map of the Cerrado with information related to the fire component. Using remotely sensed information, we identify patterns and trends of fire frequency, intensity, seasonality, extent and scar size, and combine this information for each ecoregion, relying on a simple classification that summarizes the main fire characteristics over the last two decades. Results show a marked north-south fire activity gradient, with increased contributions from MATOPIBA, the latest agricultural frontier. Five ecoregions alone account for two thirds of yearly burned area. More intense fires are found in the Arc of Deforestation and eastern ecoregions, while ecoregions in MATOPIBA display decreasing fire intensity. An innovative analysis of fire scars stratified by size class shows that infrequent large fires are responsible for the majority of burned area. These large fires display positive trends over many ecoregions, whereas smaller fires, albeit more frequent, have been decreasing in number. The final fire classification scheme shows well defined spatially-aggregated groups, where trends are found to be the key factor to evaluate fire within their regional contexts. Results presented here provide new insights to improve fire management strategies under a changing climate.

Managing fires in a changing world: Fuel and weather determine fire behavior and safety in the neotropical savannas.

Fire is an important ecological disturbance, but anthropogenic wildfires increasingly threaten native ecosystems and human lives. In fire-prone ecosystems, zero-fire policies have been replaced by active fire management to reduce the risk of wildfires and improve ecological outcomes. The environmental drivers of fire behavior are widely known, but climate change and deforestation are changing their roles, making fires less predictable. Thus, reassessing the main determinants of fire behavior is preeminent to allow for safe and adaptive uses of fire in protected areas (PA). We did this research in collaboration with PA managers during the initial implementation of a pilot Integrated Fire Management (IFM) program in the Brazilian savanna. The program mainly aimed to prevent large wildfires in the late-dry season and included prescribed burns during the rainy, early- and mid-dry seasons to create vegetation patch mosaics with different fire histories. We assessed fire behavior and its environmental drivers during prescribed fires in the mid-dry season (MF) and experimental late-dry season fires (LF) (emulating wildfires). We applied Linear Models to test for differences in fire intensity, heat released, combustion factor and flame height between fire seasons and to check the influence of meteorological and fuel conditions in these parameters. LF had a significantly higher fire intensity (3508 vs. 895 kW m-1), heat released (5537 vs. 3329 kW m-2), combustion factor (90 vs. 51%) and flame height (2.5 vs. 1.9 m) than MF. Relative humidity, air temperature, wind speed and fuel load were the best predictors of fire behavior, corroborating previous research. Air temperature and relative humidity pushed the seasonal differences in fire behavior while wind speed and fuel load showed similar effects across seasons. Our results emphasize the importance of considering primarily environmental variables during fire management planning, especially in the current climate changing world where extreme events and seasonal weather fluctuations are constantly defying our knowledge about fire behavior.

Maternal stress, low cervicovaginal ß-defensin, and spontaneous preterm birth.

Background: Spontaneous preterm birth (sPTB) is a major contributor to infant mortality and its etiology remains poorly understood. Host immunity and maternal stress may play a role in the pathogenesis of sPTB but mechanisms are poorly delineated. Antimicrobial proteins in the cervicovaginal space, such as beta defensins, modulate immune responses to bacteria and have been shown to modulate the risk of sPTB from non-optimal microbiota. While stress is known to induce immunological changes, no study has examined the interplay between maternal stress and the immune response in association with sPTB. Objectives: Our objectives were to determine whether psychosocial stress was associated with a mediator of the immune system in the cervicovaginal space, beta defensin-2, and to examine the combined impact of high stress and low cervicovaginal beta defensin-2 levels on the odds of sPTB. Study Design: From the Motherhood & Microbiome cohort study (n=2000), we performed a secondary, nested case-control study, frequency matched by race/ethnicity, of 519 pregnant women (110 sPTB and 409 term). Stress and cervicovaginal beta defensin-2 levels were measured at 16-20 weeks of gestation. Stress was dichotomized at a score of 30 on Cohen's Perceived Stress Scale (PSS-14). We measured cervicovaginal beta defensin-2 levels with ELISA and dichotomized at the median. We modeled associations of high stress and low cervicovaginal beta defensin-2 levels using multivariable logistic regression. We also compared the proportion of women with high stress and low cervicovaginal beta defensin-2 levels among women with spontaneous preterm and term births using Chi-Square tests. We modeled adjusted associations of stress and cervicovaginal beta defensin-2 levels with odds of sPTB using logistic regression. Results: The majority of the study population was non-Hispanic black (72.8%), insured by Medicaid (51.1%), and had a PSS-14 score < 30 (80.2%). High stress was associated with reduced adjusted odds of low beta defensin-2 levels (aOR 0.63, 95% CI: 0.38 -0.99). In a model adjusted for race and smoking, both high stress (aOR 1.72, 95% CI: 1.03-2.90) and low beta defensin-2 (aOR 1.58, 95% CI: 1.004-2.49) were associated with increased odds of sPTB. We then built a model of the four possible combinations of low and high stress and low and high beta defensin-2 levels with the odds of sPTB. Women with either high stress (aOR 1.37, 95% CI: 0.68 - 2.78) or low beta defensin-2 (aOR 1.40, 95% CI: 0.83-2.34), had slightly elevated but not significantly increased odds of sPTB compared to women with neither exposure. However, women with both high stress and low beta defensin-2 had significantly elevated odds of sPTB compared to women with neither exposure (aOR 3.16, 95 % CI: 1.46 - 6.84). Conclusion: High perceived stress and low cervicovaginal beta defensin-2 levels are associated with higher odds of sPTB, and when present concurrently, they result in the highest odds of sPTB in a largely non-Hispanic black cohort. Our findings warrant further work to examine social determinants of health and the host cervicovaginal immune responses that may modulate the pathogenesis of sPTB.

Comment on "The global tree restoration potential".

Bastin et al's estimate (Reports, 5 July 2019, p. 76) that tree planting for climate change mitigation could sequester 205 gigatonnes of carbon is approximately five times too large. Their analysis inflated soil organic carbon gains, failed to safeguard against warming from trees at high latitudes and elevations, and considered afforestation of savannas, grasslands, and shrublands to be restoration.

New perspectives in fire management in South American savannas: The importance of intercultural governance.

Wildfires continue to cause damage to property, livelihoods and environments around the world. Acknowledging that dealing with wildfires has to go beyond fire-fighting, governments in countries with fire-prone ecosystems have begun to recognize the multiple perspectives of landscape burning and the need to engage with local communities and their practices. In this perspective, we outline the experiences of Brazil and Venezuela, two countries where fire management has been highly contested, but where there have been recent advances in fire management approaches. Success of these new initiatives have been measured by the reduction in wildfire extent through prescribed burning, and the opening of a dialogue on fire management between government agencies and local communities. Yet, it is clear that further developments in community participation need to take place in order to avoid the appropriation of local knowledge systems by institutions, and to better reflect more equitable fire governance.

Seasonal fire management by traditional cattle ranchers prevents the spread of wildfire in the Brazilian Cerrado.

The use of fire by cattle ranchers is a major source of conflict between conservationists and local communities in tropical savannas. We evaluate the role of traditional pastoral management in wildfire prevention in two protected areas within the Brazilian savanna. Fine-grain field data from transect walks and interviews were combined with geospatial data at landscape scale to compare fire regimes in community-managed areas with those in government-managed areas. Local pastoral management creates seasonal mosaic patterns of burnings performed for productive activities and for deliberate landscape management, i.e. to protect fire-sensitive vegetation and avoid wildfires. Whereas government-managed areas were affected by large biennial late dry season wildfires, community-managed areas with a regular fire regime suffered less damage. These systems are under threat and poorly understood by researchers and environmental managers. In order to improve fire management in tropical savannas, greater understanding of pastoral management practices and their spatiotemporal dimensions is required.

The legacy of colonial fire management policies on traditional livelihoods and ecological sustainability in savannas: Impacts, consequences, new directions.

Land occupation and management systems have defined fire regimes and landscapes for millennia. The savanna biome is responsible for 86% of all fire events, contributes to 10% of the total carbon emissions annually and is home to 10% of the human population. European colonization has been associated with the implementation of fire suppression policies in many tropical savanna regions, markedly disrupting traditional fire management practices and transforming ecosystems. In this paper we assess savanna burning approaches from pre-colonial to contemporary eras in three regions: northern Australia, southern Africa and Brazil. In these regions, fire suppression policies have led to (i) conflicts between government authorities and local communities; (ii) frequent late dry season wildfires and/or (iii) woody encroachment. Such consequences are facilitating changes to fire management policies, including recognition and incorporation of traditional ecological knowledge in contemporary community-based adaptive savanna fire management. Such programs include implementation of prescribed early dry season fires and, in some regions, generating income opportunities for rural and traditional communities through the reduction of late dry season wildfires and associated greenhouse gas emissions. We present a brief history of fire management policies in these three important savanna regions, and identify ongoing challenges for implementation of culturally and ecologically sustainable fire management policies.

Genome-wide CRISPR screen for PARKIN regulators reveals transcriptional repression as a determinant of mitophagy.

PARKIN, an E3 ligase mutated in familial Parkinson's disease, promotes mitophagy by ubiquitinating mitochondrial proteins for efficient engagement of the autophagy machinery. Specifically, PARKIN-synthesized ubiquitin chains represent targets for the PINK1 kinase generating phosphoS65-ubiquitin (pUb), which constitutes the mitophagy signal. Physiological regulation of PARKIN abundance, however, and the impact on pUb accumulation are poorly understood. Using cells designed to discover physiological regulators of PARKIN abundance, we performed a pooled genome-wide CRISPR/Cas9 knockout screen. Testing identified genes individually resulted in a list of 53 positive and negative regulators. A transcriptional repressor network including THAP11 was identified and negatively regulates endogenous PARKIN abundance. RNAseq analysis revealed the PARKIN-encoding locus as a prime THAP11 target, and THAP11 CRISPR knockout in multiple cell types enhanced pUb accumulation. Thus, our work demonstrates the critical role of PARKIN abundance, identifies regulating genes, and reveals a link between transcriptional repression and mitophagy, which is also apparent in human induced pluripotent stem cell-derived neurons, a disease-relevant cell type.

Discovery and Characterization of Allosteric WNK Kinase Inhibitors.

Protein kinases are known for their highly conserved adenosine triphosphate (ATP)-binding site, rendering the discovery of selective inhibitors a major challenge. In theory, allosteric inhibitors can achieve high selectivity by targeting less conserved regions of the kinases, often with an added benefit of retaining efficacy under high physiological ATP concentration. Although often overlooked in favor of ATP-site directed approaches, performing a screen at high ATP concentration or stringent hit triaging with high ATP concentration offers conceptually simple methods of identifying inhibitors that bind outside the ATP pocket. Here, we applied the latter approach to the With-No-Lysine (K) (WNK) kinases to discover lead molecules for a next-generation antihypertensive that requires a stringent safety profile. This strategy yielded several ATP noncompetitive WNK1-4 kinase inhibitors, the optimization of which enabled cocrystallization with WNK1, revealing an allosteric binding mode consistent with the observed exquisite specificity for WNK1-4 kinases. The optimized compound inhibited rubidium uptake by sodium chloride cotransporter 1 (NKCC1) in HT29 cells, consistent with the reported physiology of WNK kinases in renal electrolyte handling.

Differentiation and visualization of diverse cellular phenotypic responses in primary high-content screening.

High-throughput screening, based on subcellular imaging, has become a powerful tool in lead discovery. Through the generation of high-quality images, not only the specific target signal can be analyzed but also phenotypic changes of the whole cell are recorded. Yet analysis strategies for the exploration of high-content screening results, in a manner that is independent from predefined control phenotypes, are largely missing. The approach presented here is based on a well-established modeling technique, self-organizing maps (SOMs), which uses multiparametric results to group treatments that create similar morphological effects. This report describes a novel visualization of the SOM clustering by using an image of the cells from each node, with the most representative cell highlighted to deploy the phenotype described by each node. The approach has the potential to identify both expected hits and novel cellular phenotypes. Moreover, different chemotypes, which cause the same phenotypic effects, are identified, thus facilitating "scaffold hopping."