Forecasting shifts in habitat suitability of three marine predators suggests a rapid decline in inter‐specific overlap under future climate change

Understanding how environmental and climate change can alter habitat overlap of marine predators has great value for the management and conservation of marine ecosystems. Here, we estimated spatiotemporal changes in habitat suitability and inter‐specific overlap among three marine predators: Baltic gray seals (Halichoerus grypus), harbor seals (Phoca vitulina), and harbor porpoises (Phocoena phocoena) under contemporary and future conditions. Location data (>200 tagged individuals) were collected in the southwestern region of the Baltic Sea; one of the fastest‐warming semi‐enclosed seas in the world. We used the maximum entropy (MaxEnt) algorithm to estimate changes in total area size and overlap of species‐specific habitat suitability between 1997–2020 and 2091–2100. Predictor variables included environmental and climate‐sensitive oceanographic conditions in the area. Sea‐level rise, sea surface temperature, and salinity data were taken from representative concentration pathways [RCPs] scenarios 6.0 and 8.5 to forecast potential climate change effects. Model output suggested that habitat suitability of Baltic gray seals will decline over space and time, driven by changes in sea surface salinity and a loss of currently available haulout sites following sea‐level rise in the future. A similar, although weaker, effect was observed for harbor seals, while suitability of habitat for harbor porpoises was predicted to increase slightly over space and time. Inter‐specific overlap in highly suitable habitats was also predicted to increase slightly under RCP scenario 6.0 when compared to contemporary conditions, but to disappear under RCP scenario 8.5. Our study suggests that marine predators in the southwestern Baltic Sea may respond differently to future climatic conditions, leading to divergent shifts in habitat suitability that are likely to decrease inter‐specific overlap over time and space. We conclude that climate change can lead to a marked redistribution of area use by marine predators in the region, which may influence local food‐web dynamics and ecosystem functioning.     

Swing‐out opening of stromal interaction molecule 1

Stromal interaction molecule 1 (STIM1) resides in the endoplasmic reticulum (ER) membrane and senses luminal calcium (Ca²⁺) concentration. STIM1 activation involves a large‐scale conformational transition that exposes a STIM1 domain termed “CAD/SOAR”, ‐ which is required for activation of the calcium channel Orai. Under resting cell conditions, STIM1 assumes a quiescent state where CAD/SOAR is suspended in an intramolecular clamp formed by the coiled‐coil 1 domain (CC1) and CAD/SOAR. Here, we present a structural model of the cytosolic part of the STIM1 resting state using molecular docking simulations that take into account previously reported interaction sites between the CC1α1 and CAD/SOAR domains. We corroborate and refine previously reported interdomain coiled‐coil contacts. Based on our model, we provide a detailed analysis of the CC1‐CAD/SOAR binding interface using molecular dynamics simulations. We find a very similar binding interface for a proposed domain‐swapped configuration of STIM1, where the CAD/SOAR domain of one monomer interacts with the CC1α1 domain of another monomer of STIM1. The rich structural and dynamical information obtained from our simulations reveals novel interaction sites such as M244, I409, or E370, which are crucial for STIM1 quiescent state stability. We tested our predictions by electrophysiological and Förster resonance energy transfer experiments on corresponding single‐point mutants. These experiments provide compelling support for the structural model of the STIM1 quiescent state reported here. Based on transitions observed in enhanced‐sampling simulations paired with an analysis of the quiescent STIM1 conformational dynamics, our work offers a first atomistic model for CC1α1‐CAD/SOAR detachment.     

Demosponge distribution in the eastern Mediterranean: a NW–SE gradient

The purpose of this paper was to investigate patterns of demosponge distribution along gradients of environmental conditions in the biogeographical subzones of the eastern Mediterranean (Aegean and Levantine Sea). The Aegean Sea was divided into six major areas on the basis of its geomorphology and bathymetry. Two areas of the Levantine Sea were additionally considered. All available data on demosponge species numbers and abundance in each area, as well as their vertical and general geographical distribution were taken from the literature. Multivariate analysis revealed a NW–SE faunal gradient, showing an apparent dissimilarity among the North Aegean, the South Aegean and the Levantine Sea, which agrees with the differences in the geographical, physicochemical and biological characteristics of the three areas. The majority of demosponge species has been recorded in the North Aegean, while the South Aegean is closer, in terms of demosponge diversity, to the oligotrophic Levantine Sea. The number of studies in the Aegean and Levantine subareas was positively correlated with the number of species recorded within each Aegean subarea. Demosponge species with an Altanto-Mediterranean distribution prevailed in the Aegean and the Levantine. The reduced contribution of the endemic component, as compared to the western Mediterranean, is consistent with the general NW–SE decrease in the number of endemic species in the Mediterranean. Demosponge distribution at the order level showed also a NW–SE gradient, similar to that observed in the entire Atlantic–Mediterranean region, suggesting a warm water affinity. Sublittoral, circalittoral and bathyal zones were clearly distinguishable in the Aegean Sea on the basis of their sponge fauna. The total number of species was an exponential negative function of depth.     

Community structure of deep-sea demersal fish in the North Aegean Sea (northeastern Mediterranean)

The spatial structure and seasonal changes of the demersal fish assemblages on the continental shelf (100–200 m) and upper slope (200–500 m) in the North Aegean Sea (Northern Aegean and Thracian Seas, northeastern Mediterranean, Greece) were analysed. Seasonal experimental trawl surveys, carried out from summer 1990 to autumn 1993, provided a total of 151 demersal fish species. Analysis of 259 bottom trawls showed the existence of four groups associated with the continental shelf and the upper slope; each group was dominated by a small number of species. The bathymetric distribution of the species, established using measures of the centre of gravity and habitat width, revealed that most of the species had a wide distributional range within the study area, although a few were restricted to the greatest depths. Density, biomass, species richness and diversity decreased significantly with depth, and were also indicative of distinctive characteristics between these fish assemblages. Mean fish weight exhibited two different trends: a bigger-deeper phenomenon at the continental shelf and a smaller-deeper phenomenon at the upper slope. The variability in assemblage structure was determined mainly by depth and, to a lesser extent, by season and geographical location. For some species, results suggest a pattern of gradual species replacement along the depth gradient coupled with ontogenetic habitat shifts.     

Habitat Selection Response of Small Pelagic Fish in Different Environments. Two Examples from the Oligotrophic Mediterranean Sea

A number of scientific papers in the last few years singled out the influence of environmental conditions on the spatial distribution of fish species, highlighting the need for the fisheries scientific community to investigate, besides biomass estimates, also the habitat selection of commercially important fish species. The Mediterranean Sea, although generally oligotrophic, is characterized by high habitat variability and represents an ideal study area to investigate the adaptive behavior of small pelagics under different environmental conditions. In this study the habitat selection of European anchovy Engraulis encrasicolus and European sardine Sardina pilchardus is analyzed in two areas of the Mediterranean Sea that largely differentiate in terms of environmental regimes: the Strait of Sicily and the North Aegean Sea. A number of environmental parameters were used to investigate factors influencing anchovy and sardine habitat selection. Acoustic surveys data, collected during the summer period 2002–2010, were used for this purpose. The quotient analysis was used to identify the association between high density values and environmental variables; it was applied to the entire dataset in each area in order to identify similarities or differences in the “mean” spatial behavioral pattern for each species. Principal component analysis was applied to selected environmental variables in order to identify those environmental regimes which drive each of the two ecosystems. The analysis revealed the effect of food availability along with bottom depth selection on the spatial distribution of both species. Furthermore PCA results highlighted that observed selectivity for shallower waters is mainly associated to specific environmental processes that locally increase productivity. The common trends in habitat selection of the two species, as observed in the two regions although they present marked differences in hydrodynamics, seem to be driven by the oligotrophic character of the study areas, highlighting the role of areas where the local environmental regimes meet ‘the ocean triad hypothesis’.     

Cryo‐EM structure of native human uromodulin,a zona pellucida module polymer

Assembly of extracellular filaments and matrices mediating fundamental biological processes such as morphogenesis, hearing, fertilization, and antibacterial defense is driven by a ubiquitous polymerization module known as zona pellucida (ZP) “domain”. Despite the conservation of this element from hydra to humans, no detailed information is available on the filamentous conformation of any ZP module protein. Here, we report a cryo‐electron microscopy study of uromodulin (UMOD)/Tamm–Horsfall protein, the most abundant protein in human urine and an archetypal ZP module‐containing molecule, in its mature homopolymeric state. UMOD forms a one‐start helix with an unprecedented 180‐degree twist between subunits enfolded by interdomain linkers that have completely reorganized as a result of propeptide dissociation. Lateral interaction between filaments in the urine generates sheets exposing a checkerboard of binding sites to capture uropathogenic bacteria, and UMOD‐based models of heteromeric vertebrate egg coat filaments identify a common sperm‐binding region at the interface between subunits.     

Molecular insights into the primary nucleation of polymorphic amyloid β dimers in DOPC lipid bilayer membrane

Alzheimer''s disease (AD) pathology is characterized by loss of memory cognitive and behavioral deterioration. One of the hallmarks of AD is amyloid β (Aβ) plaques in the brain that consists of Aβ oligomers and fibrils. It is accepted that oligomers, particularly dimers, are toxic species that are produced extracellularly and intracellularly in membranes. It is believed that the disruption of membranes by polymorphic Aβ oligomers is the key for the pathology of AD. This is a first study that investigate the effect of polymorphic “α‐helix/random coil” and “fibril‐like” Aβ dimers on 1,2‐dioleoyl‐sn‐glycero‐3‐phosphocholine (DOPC) membrane. It has been found that the DOPC membrane promotes Aβ_1–42 “fibril‐like” dimers and impedes Aβ_1–42 “α‐helix/random coil” dimers. The N‐termini domains within Aβ_1–42 dimers play a role in Aβ aggregation in membrane milieus. In addition, the aromatic π–π interactions (involving residues F19 and F20 in Aβ_1–42) are the driving forces for the hydrophobic interactions that initiate the primary nucleation of polymorphic Aβ_1–42 dimers within DOPC membrane. Finally, the DOPC bilayer membrane thickness is locally decreased, and it is disrupted by an embedded distinct Aβ_1–42 dimer, due to relatively large contacts between Aβ_1–42 monomers and the DOPC membrane. This study reveals insights into the molecular mechanisms by which polymorphic early‐stage Aβ_1–42 dimers have distinct impacts on DOPC membrane.     

A multi‐data ensemble approach for predicting woodland type distribution: Oak woodland in Britain

Interactions between soil, topography, and climatic site factors can exacerbate and/or alleviate the vulnerability of oak woodland to climate change. Reducing climate‐related impacts on oak woodland habitats and ecosystems through adaptation management requires knowledge of different site interactions in relation to species tolerance. In Britain, the required thematic detail of woodland type is unavailable from digital maps. A species distribution model (SDM) ensemble, using biomod2 algorithms, was used to predict oak woodland. The model was cross‐validated (50%:50% ‐ training:testing) 30 times, with each of 15 random sets of absence data, matching the size of presence data, to maximize environmental variation while maintaining data prevalence. Four biomod2 algorithms provided stable and consistent TSS‐weighted ensemble mean results predicting oak woodland as a probability raster. Biophysical data from the Ecological Site Classification (forest site classification) for Britain were used to characterize oak woodland sites. Several forest datasets were used, each with merits and weaknesses: public forest estate subcompartment database map (PFE map) for oak‐stand locations as a training dataset; the national forest inventory (NFI) “published regional reports” of oak woodland area; and an “NFI map” of indicative forest type broad habitat. Broadleaved woodland polygons of the NFI map were filled with the biomod2 oak woodland probability raster. Ranked pixels were selected up to the published NFI regional area estimate of oak woodland and matched to the elevation distribution of oak woodland stands, from “NFI survey” sample squares. Validation using separate oak woodland data showed that the elevation filter significantly improved the accuracy of predictions from 55% (p = .53) to 83% coincidence success rate (p < .0001). The biomod2 ensemble, with masking and filtering, produced a predicted oak woodland map, from which site characteristics will be used in climate change interaction studies, supporting adaptation management recommendations for forest policy and practice.     

Historical anthropogenic disturbances explain long‐term moorland vegetation dynamics

Upland moorlands are important landscapes, but many are considered degraded as a result of human activities. Consequently, their protection and restoration are of substantial concern. In Europe, restoration activities are often aimed at reversing the effects of 19th and 20th century “agricultural improvements,” which often involved major drainage schemes. However, the ecological effects and long‐term ecological context of “agricultural improvement” are not yet fully understood. To develop this understanding, we analyze paleoecological data (pollen, coprophilous fungal spores, microcharcoal) from five upland peatland sites using a range of analytical approaches: cluster analysis, principal component analysis, rate‐of‐change analysis, and regression analyses incorporating documentary historical data. The sites are located on Exmoor (South West England, UK), a landscape that typifies historic upland degradation. We demonstrate that in this landscape, 19th century drainage is associated with declines in Sphagnum and non‐arboreal taxon richness; over longer timescales burning is associated with enhanced graminoid monocot abundance and grazing with lower taxon richness. We also show that rate‐of‐change in moorland vegetation communities during the 19th century is not distinctive in a long‐term context: change has been a constant in this landscape, rather than an exception during the 19th century. Our findings indicate that the aims of “restoration” interventions intended to increase Sphagnum abundances, increase taxon richness and reduce graminoid dominance are consistent with the long‐term dynamics of peatland systems, such as those on Exmoor. “Restoration” deemed successful in these terms may or may not resemble pre‐drainage conditions, which were themselves a function of millennia of successive moorland management regimes.     

Biosafety in DIY‐bio laboratories: from hype to policy: Discussions about regulating DIY biology tend to ignore the extent of self‐regulation and oversight of DIY laboratories

Policymakers should treat DIY‐biology laboratories as legitimate parts of the scientific enterprise and pay attention to the role of community norms. Subject Categories: Synthetic Biology & Biotechnology, S&S: Economics & Business, S&S: Ethics

DIY biology – very broadly construed as the practice of biological experiments outside of traditional research environments such as universities, research institutes or companies – has, during the past decade, gained much prominence. This increased attention has raised a number of questions about biosafety and biosecurity, both in the media and by policy makers who are concerned about safety and security lapses in “garage biology”. There are a number of challenges here though when it comes to policies to regulate DIY biology. For a start, the term itself escapes easy definition: synonyms or related terms abound, including garage biotechnology, bio‐hacking, self‐modification/grinding, citizen science, bio‐tinkering, bio‐punk, even transhumanism. Some accounts even use ‘DIY‐bio’ interchangeably with synthetic biology, even though these terms refer to different emerging trends in biology. Some of these terms are more charged than others but each carries its own connotations with regard to practice, norms and legality. As such, conversations about the risk, safety and regulation of DIY‐bio can be fraught.

Synonyms or related terms abound, including garage biotechnology, bio‐hacking, self‐modification/grinding, citizen science, bio‐tinkering, bio‐punk, even transhumanism.

Given the increasing policy discussions about DIY‐bio, it is crucial to consider prevailing practice thoughtfully, and accurately. Key questions that researchers, policy makers and the public need to contemplate include the following: “How do different DIY‐bio spaces exist within regulatory frameworks, and enact cultures of (bio)safety?”, “How are these influenced by norms and governance structures?”, “If something is unregulated, must it follow that it is unsafe?” and “What about the reverse: does regulatory oversight necessarily lead to safer practice?”.The DIY‐bio movement emerged from the convergence of two trends in science and technology. The first one is synthetic biology, which can broadly be defined as a conception of genetic engineering as systematic, modular and programmable. While engineering living organisms is obviously a complex endeavour, synthetic biology has sought to re‐frame it by treating genetic components as inherently modular pieces to be assembled, through rational design processes, into complex but predictable systems. This has prompted many “LEGO” metaphors and a widespread sense of democratisation, making genetic engineering accessible not only to trained geneticists, but also to anyone with an “engineering mindset”.The second, much older, trend stems from hacker‐ and makerspaces, which are – usually not‐for‐profit – community organisations that enable groups of enthusiasts to share expensive or technically complex infrastructure, such as 3D printers or woodworking tools, for their projects. These provide a model of community‐led initiatives based on the sharing of infrastructure, equipment and knowledge. Underpinning these two trends is an economic aspect. Many of the tools of synthetic biology – notably DNA sequencing and synthesis – have seen a dramatic drop in cost, and much of the necessary physical apparatus is available for purchase, often second‐hand, through auction sites.DIY‐bio labs are often set‐up under widely varying management schemes. While some present themselves as community outreach labs focusing on amateur users, others cater specifically to semi‐ or professional members with advanced degrees in the biosciences. Other such spaces act as incubators for biotech startups with an explicitly entrepreneurial culture. Membership agreements, IP arrangements, fees, access and the types of project that are encouraged in each of these spaces can have a profound effect on the science being done.     

Phenotypic plasticity in life‐history traits of Daphnia galeata in response to temperature – a comparison across clonal lineages separated in time

Climatic changes are projected to result in rapid adaptive events with considerable phenotypic shifts. In order to reconstruct the impact of increased mean water temperatures during past decades and to reveal possible thermal micro‐evolution, we applied a resurrection ecology approach using dormant eggs of the freshwater keystone species Daphnia galeata. To this end, we compared the adaptive response of D. galeata clones from Lake Constance of two different time periods, 1965–1974 (“historical”) versus 2000–2009 (“recent”), to experimentally increased temperature regimes. In order to distinguish between genetic versus environmentally induced effects, we performed a common garden experiment in a flow‐through system and measured variation in life‐history traits. Experimental thermal regimes were chosen according to natural temperature conditions during the reproductive period of D. galeata in Central European lakes, with one additional temperature regime exceeding the currently observable maximum (+2°C). Increased water temperatures were shown to significantly affect measured life‐history traits, and significant “temperature × clonal age” interactions were revealed. Compared to historical clones, recent clonal lineages exhibited a shorter time to first reproduction and a higher survival rate, which may suggest temperature‐driven micro‐evolution over time but does not allow an explicit conclusion on the adaptive nature of such responses.     

Modelling potential habitat of the invasive ctenophore <Emphasis Type="Italic">Mnemiopsis leidyi</Emphasis> in Aegean Sea

The invasive ctenophore Mnemiopsis leidyi was accidentally introduced into the Black Sea in the early 1980s and it was first sighted in the Aegean Sea (Eastern Mediterranean) in the early 1990s. This article presents a first attempt to develop a predictive spatial model based on M. leidyi presence data and satellite environmental data from the Aegean Sea during early summer, in order to identify those areas in the Greek Seas and the entire Mediterranean basin that could serve as potential habitat for the species. Generalized additive models (GAM) were applied. The final GAM model indicated higher probability of finding M. leidyi present in depths of 65–135 m and sea surface temperature values of 21–25°C. Furthermore, the significant interaction between photosynthetically active radiation (PAR) and sea level anomaly (SLA) indicated a higher probability of M. leidyi presence in low values of PAR and SLA. In the next step, the final GAM was applied in a prediction grid of mean monthly satellite values for June 2004–2006 in order to estimate probability of M. leidyi presence in the Hellenic Seas and the whole Mediterranean basin at a GIS resolution of 4 km. In the Aegean Sea, species potential habitat included areas influenced by the Black Sea Water (e.g. Thracian Sea, Limnos-Imvros plateau), gulfs that are affected by river runoffs, such as the Thermaikos, Strymonikos and Patraikos gulfs, or areas with strong anthropogenic influence such as the Saronikos gulf. Areas with the same environmental conditions as those in Aegean Sea have been indicated in certain spots of the Levantine Sea as well as in coastal waters of Egypt and Libya, although their spatial extent varied largely among years examined. However, the occurrence of conditions that are linked to high probability of M. leidyi presence does not necessarily mean that these areas can support successful reproduction, high population or bloom levels, since these depend on a combination of temperature, salinity, food availability and the abundance of predators. Guest editor: V. D. Valavanis Essential Fish Habitat Mapping in the Mediterranean     

Exercise‐induced α‐ketoglutaric acid stimulates muscle hypertrophy and fat loss through OXGR1‐dependent adrenal activation

The authors approached the journal to correct a mistake in the data presented in Appendix␣Fig S3D. The authors state that the mouse images in Appendix␣Fig S3D mistakenly displayed images from Fig 2F and Appendix␣Fig S1F. The images in Appendix␣Fig S3D are herewith corrected. The authors state that this change does not affect the conclusions or the statistics. The source data for these panels have been added to the original publication.The authors note that the following sentence needs to be corrected from: Appendix Figure S3D. Original. Appendix Figure S3D. Corrected. “Interestingly, several well‐established accumulation signatures of succinate, malate, hypoxanthine, and xanthine induced by endurance exercise (Lewis et␣al, 2010) were found to be decreased by endurance exercise (Figs 1D and EV1A–D)”.to“Interestingly, several well‐established accumulation signatures of succinate, malate, hypoxanthine, and xanthine induced by endurance exercise (Lewis et␣al, 2010) were found to be decreased by resistance exercise (Figs 1D and EV1A–D)”.Further, the authors requested to amend the legend of Appendix␣Fig S3R to indicate that the same sample for the iWAT group, “WT+2%AKG” treatment, is shown in Fig 3P. The corrected legend reads: “(R‐S). Representative images (R) and quantification (S) of p‐HSL DAB staining from male OXGR1OE^AG mice treated with AKG for 12 weeks (n = 6 per group). The same sample is shown as in Fig 3P ”.The authors regret these errors and any confusion they may have caused. All authors approve of this correction.     

Shifts in fisheries management: adapting to regime shifts

For many years, fisheries management was based on optimizing yield and maintaining a target biomass, with little regard given to low-frequency environmental forcing. However, this policy was often unsuccessful. In the last two to three decades, fisheries science and management have undergone a shift towards balancing sustainable yield with conservation, with the goal of including ecosystem considerations in decision-making frameworks. Scientific understanding of low-frequency climate–ocean variability, which is manifested as ecosystem regime shifts and states, has led to attempts to incorporate these shifts and states into fisheries assessment and management. To date, operationalizing these attempts to provide tactical advice has met with limited success. We review efforts to incorporate regime shifts and states into the assessment and management of fisheries resources, propose directions for future investigation and outline a potential framework to include regime shifts and changes in ecosystem states into fisheries management.     

Demographic responses to climate‐driven variation in habitat quality across the annual cycle of a migratory bird species

The demography and dynamics of migratory bird populations depend on patterns of movement and habitat quality across the annual cycle. We leveraged archival GPS‐tagging data, climate data, remote‐sensed vegetation data, and bird‐banding data to better understand the dynamics of black‐headed grosbeak (Pheucticus melanocephalus) populations in two breeding regions, the coast and Central Valley of California (Coastal California) and the Sierra Nevada mountain range (Sierra Nevada), over 28 years (1992–2019). Drought conditions across the annual cycle and rainfall timing on the molting grounds influenced seasonal habitat characteristics, including vegetation greenness and phenology (maturity dates). We developed a novel integrated population model with population state informed by adult capture data, recruitment rates informed by age‐specific capture data and climate covariates, and survival rates informed by adult capture–mark–recapture data and climate covariates. Population size was relatively variable among years for Coastal California, where numbers of recruits and survivors were positively correlated, and years of population increase were largely driven by recruitment. In the Sierra Nevada, population size was more consistent and showed stronger evidence of population regulation (numbers of recruits and survivors negatively correlated). Neither region showed evidence of long‐term population trend. We found only weak support for most climate–demographic rate relationships. However, recruitment rates for the Coastal California region were higher when rainfall was relatively early on the molting grounds and when wintering grounds were relatively cool and wet. We suggest that our approach of integrating movement, climate, and demographic data within a novel modeling framework can provide a useful method for better understanding the dynamics of broadly distributed migratory species.     

Genetic association study of intron variants in the forkhead box protein P3 gene in Chinese patients diagnosed with cervical cancer

The aim of this study was to investigate the effects of forkhead box protein P3 (FOXP3) intron single nucleotide variants (SNVs) in high‐risk human papilloma virus (HR‐HPV) infection and cervical cancer (CC) malignant lesions. We performed FOXP3 genotyping in 350 patients with CC and 350 healthy controls using the ImLDR multiple single nucleotide polymorphism genotyping technology. The heterozygous mutation TC in rs2294021 decreased the risk of HR‐HPV infection and CC malignant lesions (TC vs. TT: OR = 0.71, 95% CI = 0.51–0.99); the dominant model TC+CC and allele C in rs2294021 decreased the risk of CC malignant lesions (TC+CC vs. TT: OR = 0.69, 95% CI = 0.50–0.95; C vs. T: OR = 0.78, 95% CI = 0.63–0.97). The heterozygous mutation GA, dominant model GA+AA and allele A in rs3761549 also decreased the risk of HR‐HPV infection and CC malignant lesions (GA vs. GG: OR = 0.70, 95% CI = 0.51–0.96; GA+AA vs. GG: OR = 0.69, 95% CI = 0.51–0.94; A vs. G: OR = 0.75, 95% CI = 0.58–0.96). Patients with CC and HR‐HPV infection carrying rs2294021 TC and rs3761549 GA had lower expression of FOXP3 protein. Haplotype analysis revealed that T‐C‐A decreased the risk of HR‐HPV infection. Furthermore, we found a significant association between immune cells infiltration and prognosis in patients with CC. Our findings demonstrated that rs2294021 and rs3761549 variants may protect against HR‐HPV and CC malignant lesions by downregulating FOXP3 and that FOXP3 was associated with immune cells infiltration, which affected the prognosis of CC.     

NLRP3 activation contributes to endothelin‐1‐induced erectile dysfunction

In the present study, we hypothesized that endothelin (ET) receptors (ET_A and ET_B) stimulation, through increased calcium and ROS formation, leads to Nucleotide Oligomerization Domain‐Like Receptor Family, Pyrin Domain Containing 3 (NLRP3) activation. Intracavernosal pressure (ICP/MAP) was measured in C57BL/6 (WT) mice. Functional and immunoblotting assays were performed in corpora cavernosa (CC) strips from WT, NLRP3^−/− and caspase^−/− mice in the presence of ET‐1 (100 nM) and vehicle, MCC950, tiron, BAPTA AM, BQ123, or BQ788. ET‐1 reduced the ICP/MAP in WT mice, and MCC950 prevented the ET‐1 effect. ET‐1 decreased CC ACh‐, sodium nitroprusside (SNP)‐induced relaxation, and increased caspase‐1 expression. BQ123 an ET_A receptor antagonist reversed the effect. The ET_B receptor antagonist BQ788 also reversed ET‐1 inhibition of ACh and SNP relaxation. Additionally, tiron, BAPTA AM, and NLRP3 genetic deletion prevented the ET‐1‐induced loss of ACh and SNP relaxation. Moreover, BQ123 diminished CC caspase‐1 expression, while BQ788 increased caspase‐1 and IL‐1β levels in a concentration‐dependent manner (100 nM–10 μM). Furthermore, tiron and BAPTA AM prevented ET‐1‐induced increase in caspase‐1. In addition, BAPTA AM blocked ET‐1‐induced ROS generation. In conclusion, ET‐1‐induced erectile dysfunction depends on ET_A‐ and ET_B‐mediated activation of NLRP3 in mouse CC via Ca²⁺‐dependent ROS generation.     

Neutralization of SARS‐CoV‐2 by highly potent,hyperthermostable, and mutation‐tolerant nanobodies

Monoclonal anti‐SARS‐CoV‐2 immunoglobulins represent a treatment option for COVID‐19. However, their production in mammalian cells is not scalable to meet the global demand. Single‐domain (VHH) antibodies (also called nanobodies) provide an alternative suitable for microbial production. Using alpaca immune libraries against the receptor‐binding domain (RBD) of the SARS‐CoV‐2 Spike protein, we isolated 45 infection‐blocking VHH antibodies. These include nanobodies that can withstand 95°C. The most effective VHH antibody neutralizes SARS‐CoV‐2 at 17–50 pM concentration (0.2–0.7 µg per liter), binds the open and closed states of the Spike, and shows a tight RBD interaction in the X‐ray and cryo‐EM structures. The best VHH trimers neutralize even at 40 ng per liter. We constructed nanobody tandems and identified nanobody monomers that tolerate the K417N/T, E484K, N501Y, and L452R immune‐escape mutations found in the Alpha, Beta, Gamma, Epsilon, Iota, and Delta/Kappa lineages. We also demonstrate neutralization of the Beta strain at low‐picomolar VHH concentrations. We further discovered VHH antibodies that enforce native folding of the RBD in the E. coli cytosol, where its folding normally fails. Such “fold‐promoting” nanobodies may allow for simplified production of vaccines and their adaptation to viral escape‐mutations.