Biology Open

UNC-6/Netrin is a conserved regulator of dorsal-ventral axon and cell migrations. UNC-6 is composed of a Laminin N-terminal domain (LN), three epidermal growth factor repeats (EGF), and a Netrin C terminal domain (NC). Here, we identified missense mutations in distinct UNC-6 domains and assessed their roles in dorsal VD/DD motor axon guidance and ventral AVM axon guidance. A missense mutation in a conserved residue of the LN domain (G289D) resulted in dorsal and ventral axon guidance defects similar to unc-6 null. A distinct missense mutation in the LN domain (S120F) was hypomorphic and strongly perturbed ventral AVM axon guidance with minimal effects on dorsal VD/DD axon guidance, showing that S120F is predominantly required for ventral guidance. Missense mutations altering conserved cysteine residues involved in di-sulfide bonding in the EGF domains were analyzed. EGF1(C321G) caused both ventral and dorsal axon guidance defects albeit weaker than unc-6 null, indicating that EGF1 is required for both. EGF2(C347Y) strongly affected dorsal VD/DD axon guidance similar to unc-6 null, with weaker perturbation of ventral AVM axon guidance. Previous results revealed that EGF3(C410Y) specifically disrupted dorsal axon guidance, a result that we confirmed. Our studies using missense mutations in the endogenous unc-6 locus complement previous structure-function studies using transgenic expression, and identify domains specifically required for ventral AVM guidance (S120Y in the LN domain) and dorsal VD/DD axon guidance (C410Y in EGF3). The crystal structure of UNC-6 indicates conserved N-linked glycosylation at N114 and N128. Mutation of these sites in UNC-6 had no effect on dorsal ventral axon guidance, showing that they do not play a major role. However, the N114 and N128 mutations interacted genetically with unc-40 and unc-5 mutations, indicating that these glycosylation sites indeed have a role in UNC-6 signaling. Our results will inform studies on how these distinct UNC-6 domains interact with guidance receptors (e.g. UNC-40/DCC and UNC-5) and other extracellular molecules to mediate dorsal-ventral axon guidance.

Microcomputed tomography (micro-CT) is a useful tool that can be utilised for 3D structural characterisation and volumetric quantification of small biological specimens. Its potential application is particularly valuable within the field of cardiac development, where phenotypic profiling at the whole organ, cell, and molecular level is often most informative within the same sample. Consequently, this study sought to develop a multimodal imaging protocol to enable 3D phenotypic characterisation of embryonic avian hearts (iodine-based contrast X-ray imaging) prior to immunohistochemistry-based cell and molecular analysis. Micro-CT parameters were tested to establish an optimal protocol for 3D analysis of embryonic cardiac specimens across multiple developmental timepoints. Optimised parameters provided reliable and reproducible 3D analysis of cardiac macrostructures. Sodium thiosulphate treatment of X-ray imaged hearts effectively reversed the iodine-based contrast stain whilst maintaining antigen availability of nuclear, membranous, and cytoplasmic targets in traditional downstream imaging studies. Together, this study demonstrates a robust and highly efficient multimodal imaging strategy to comprehensively characterise cardiac morphology in avian embryos and may serve as a versatile foundation for a broad range of bioimaging applications within the wider scientific community.

Motility of the malaria-causing parasite Plasmodium is essential for transmission to and from mosquitoes, with the turnover of actin filaments being a central feature of productive cell movement. Actin-related proteins (Arps) are known to play critical roles in motility, trafficking and chromatin remodelling. Here, we show that the actin-like protein 1 (Alp1), an apicomplexan Arp, is essential for Plasmodium ookinete motility and establishing of infection in mosquitoes. We identified an insertion region in subdomain 4 that contributes to Alp1 function in ookinetes and show novel actin filament structures in ookinetes. A combination of gene deletion and actin filament recognizing chromobody expression revealed a role of Alp1 in promoting actin filament turnover in ookinetes. We have thus identified a novel Arp that has evolved a specialist function to regulate actin dynamics, govern malaria parasite motility and facilitate malaria transmission.

ABSTRACT Background: Athletes who return to soccer after anterior cruciate ligament reconstruction (ACLR) remain at elevated risk of secondary injury despite meeting conventional discharge criteria, and neuromuscular deficits in the reconstructed limb are known to be exposed by fatigue. Objective: To determine whether match-play fatigue differentially affects muscle stiffness, countermovement jump (CMJ) force symmetry, and rate of force development (RFD) asymmetry between soccer players with a history of ACLR and uninjured teammates. Methods: A prospective, cross-sectional, matched-control study enrolled 128 competitive soccer players (64 ACLR, 6-22 months post-surgery; 64 uninjured controls) across five recruitment waves (February-June 2026). Bilateral CMJ peak vertical force, jump height, RFD, and myotonometric stiffness of the rectus femoris (RF), vastus medialis (VM), and biceps femoris (BF) were recorded immediately before and after a standardized competitive match. Fatigue was quantified from second-half heart rate (percentage of age-predicted maximum) and end-match rating of perceived exertion (RPE). Within-group pre-to-post changes were evaluated with paired t-tests, between-group differences in the magnitude of change with independent-samples t-tests, and associations between fatigue indices and asymmetry changes with Pearson correlations. Results: Match play reduced CMJ limb symmetry index (LSI) in both groups, but the decline was more than three-fold greater in the ACLR group, 92.6% (SD 5.4%) to 85.1% (SD 7.1%), than in control group, 97.3% (SD 3.9%) to 95.0% (SD 4.2%), group-by-time difference, p < 0.001, (d = 0.64). RFD asymmetry approximately doubled in the ACLR group, 10.6% (SD 4.1%) to 17.6% (SD 6.5%), compared with a smaller rise in control group, 4.6% (SD 2.4%) to 6.3% (SD 3.7%); p < 0.001, d = 0.77). Involved-limb stiffness losses in the ACLR group exceeded those of controls for the RF (-21.2 vs. -9.2 N/m, p < 0.001), VM (-17.7 vs. -6.1 N/m, p < 0.001), and BF (-13.3 vs. -6.6 N/m, p < 0.001), whereas uninvolved-limb stiffness losses did not differ between groups (all p > 0.05). Fatigue markers (heart rate, RPE) were not significantly correlated with the magnitude of individual asymmetry change (|r| [&le;] 0.18, p > 0.15). Conclusions: In competitive soccer players 6-22 months after ACLR, match-play fatigue selectively compromises stiffness and explosive force output of the reconstructed limb, widening inter-limb asymmetries beyond what is seen in uninjured teammates, even though global cardiovascular and perceptual fatigue were comparable between groups. These findings suggest that return-to-sport testing performed only in a rested state may underestimate residual neuromuscular deficits, and support fatigue-inclusive assessment protocols before athletes are cleared for unrestricted competition. Abbreviations: ACL: anterior cruciate ligament, ACLR: anterior cruciate ligament reconstruction, BF: biceps femoris, CMJ: countermovement jump, HRmax: maximum heart rate, LSI: limb symmetry index, RF: rectus femoris, RFD: rate of force development, RPE: rating of perceived exertion, RTS: return to sport, VM: vastus medialis, SD: standard deviation. Keywords: Anterior cruciate ligament reconstruction, muscle fatigue, muscle stiffness, countermovement jump, limb symmetry index, rate of force development, soccer, return to sport.

Extensive experiments document that SPARCL1, a secreted protein that is produced primarily by astrocytes in brain and endothelia throughout the body and that is also known as Hevin, enhances synapse formation. However, the mode of action of SPARCL1 at synapses remains unclear owing to divergent results in the literature. Here, we use cultured neurons from newborn male and female mouse embryos to show that the C-terminal follistatin-like and Ca2+-binding domains of SPARCL1, which account for only 35% of the total SPARCL1 sequence, are sufficient to potently enhance synapse numbers. SPARCL1 acts at nanomolar concentrations at which SPARCL1 does not robustly bind to neurexins, neuroligins or neurexin/neuroligin complexes but avidly interacts with all teneurins. Strikingly, the follistatin-like domain of SPARCL1 on its own strongly binds to teneurins but is unable to stimulate synapse formation. Only when combined with the SPARCL1 Ca2+- binding domain does the follistatin-like domain induce synapses, suggesting that SPARCL1 enhances synapse numbers by binding to teneurins via its C-terminal follistatin-like domain and by activating synapse formation via its Ca2+-binding domain. SIGNIFICANCE STATEMENTSPARCL1 (also known as Hevin) is a synaptogenic factor that is produced primarily by astrocytes in brain, and that enhances synapse formation. How SPARCL1 acts at synapses, however, remains unclear because divergent results describe its binding partners at synapses and the sequences involved in its synaptogenic activity remain unclear. In the present study, we show that SPARCL1 avidly binds to the presynaptic teneurins adhesion molecules, that this binding is mediated by its small follistatin-like domain, and that its synaptogenic activity requires both its follistatin-like and its Ca2+-binding EC domains. Thus, our results suggest that SPARCL1 is recruited to developing synapses by binding of its follistatin-like domain to teneurins and then induces synapse assembly via its Ca2+-binding domain.

Stem cells proliferate rapidly to maintain fast tissue turnover during regeneration. However, the feedback mechanisms in stem cells that prevent hyperproliferation remain unclear, and their dysregulation can lead to organ failure and cancer. Here, we identified nuclear factor-Y (NF-Y) as the transcriptional repressors to maintain the stem cell quiescence during intestinal homeostasis. We found that NF-Y negatively regulates intestinal stem cell (ISC) proliferation through preferentially occupying the promoters of EGFR signaling pathway components Egfr/Mkp3/Raf/Ras/pointed, via the action of histone acetyltransferase Nejire (Nej)/p300 dependent transcription regulation. While the loss of NF-Y enhances ISC proliferation, cell death and sensitivity to stress and tumor induced mortality. Moreover, NF-Y acts together with Nej to restrict Egfr expression and suppress ISC hyperproliferation. Together, these results demonstrate NF-Y acts with Nej serve as a key negative feedback module to orchestrate transcription initiation and termination of growth signaling in the control of stem cell activity in homeostatic and disease conditions.

Animal cell-cultured ingredients are entering the EU and UK pet food markets under frameworks that do not require pre-market, ingredient-level safety assessments, creating an ethical need for transparent safety disclosure. We present the first public safety dossier for this sector, describing the proprietary mouse embryonic stem cell line PE25 and its derived, non-viable cellular and conditioned media ingredient produced in food and feed-grade media. PE25 characterization confirmed Mus musculus identity, sterility, absence of mycoplasma and replication-competent retroviruses, and stable growth. Doxorubicin-induced p53 stress testing, CD44/BMI1 profiling, and soft agar assays showed no cancer-like traits and a non-tumorigenic profile; the final ingredient contains no viable cells. Independent OECD TG 471 and 487 assays confirmed non-genotoxicity. Heavy metals, biogenic amines, solvents, and chemical residues were below regulatory limits. Given process variability, we recommend case-by-case safety evaluation and propose this dossier as a model for responsible commercialization.

Background: Lead exposure is estimated to cause approximately 3.5 million premature deaths a year, yet the key ongoing sources of lead exposure are unclear. Methods: We estimated the contribution of dietary lead intake to global blood lead levels (BLLs) for 7-year-old children and 22-year-old adults by applying the All-Ages Lead Model (AALM) to calculate blood lead levels (BLLs) based on 25 total diet studies (TDS) that quantify dietary lead intake across 46 countries. Results: For children, the population-weighted average dietary lead intake in low- and middle-income countries (LMICs) (32.0 g/day) was found to be more than three times higher than in high-income countries (HICs) (9.3 g/day), and more than 10 times higher than the FDA reference level for children (2.2 g/day). The average impact on BLLs for children is estimated to be near 29 g/L in LMICs and near 12 g/L in HICs. Averaged across the TDS data, vegetables (27%) and cereals (24%) were found to contribute the most to dietary lead. Conclusions: While there are limitations associated with biokinetic modelling and the TDS data from LMICs, these results suggest that the contribution of dietary lead intake to global lead exposure is in the region of 40 to 50%, suggesting, in turn, that dietary lead intake is likely a major global driver of lead poisoning. Lead absorbed from the environment into food crops is expected to be the key driver of dietary lead. Current regulatory levels for maximum lead concentrations in foods (0.05-0.3 mg/kg) are out-of-date and may imply a dietary lead intake of 200 g/day, far higher than the FDA reference level (2.2 g/day). Collecting representative TDS data in high lead burden countries should be a priority. Further research is also recommended on upstream lead sources and pathways of lead uptake in plants, driving global food contamination.

Humans and other animals often act in environments that are at least partly familiar, where aspects of the spatial layout are known. Although such knowledge is known to support navigation and spatial cognition, its role in the online control of action remains unclear. We investigated whether drivers use knowledge of road layout to guide steering in high and low visibility and, if so, the form of such knowledge. In two simulated driving experiments (total N = 90), participants repeatedly drove winding roads containing segments with and without fog. Drivers who repeatedly experienced the same road exhibited more stable steering and lane positioning than drivers encountering novel roads, but only when visibility was reduced. These advantages were accompanied by superior performance on post-tests assessing knowledge of road geometry. We next examined the form of such knowledge by dissociating global knowledge of road layout from local associations between landmarks and road segments. Disrupting landmark-road segment associations produced the largest impairment in steering performance. The benefits of prior experience were largely preserved when road-segment order was scrambled but landmark associations remained intact. These findings show that spatial knowledge can support moment-to-moment steering control when visibility is reduced. Rather than relying on a globally coherent representation of the environment, drivers use local associations between landmarks and upcoming road geometry to anticipate future demands. More broadly, the results elucidate how familiarity with environmental structure contributes to the control of action when visual information is degraded, revealing a close interplay between spatial knowledge and visual control. Significance StatementPeople routinely act within surroundings they have encountered before, from commuting on the same streets to walking familiar hallways. Whether the spatial knowledge acquired from such experience actually shapes online visual control remains an open question. Using a simulated driving task, we show that familiarity with a road improves steering stability specifically when visibility is reduced, and that this benefit depends on learned associations between landmarks and upcoming road geometry rather than a global cognitive map. The results indicate that spatial knowledge plays a key role in moment-to-moment control, letting drivers anticipate road segments they cannot yet see. Unfamiliar roads and impaired spatial learning may compound the risks of poor visibility, suggesting a role for driver-assistance systems that leverage landmarks.

Efferocytosis, the recognition, engulfment, and degradation of apoptotic cells by phagocytes, is essential for tissue homeostasis and development, and its failure contributes to chronic inflammation and neurodegeneration. The amyloid precursor protein (APP), a central pathogenic factor in Alzheimers disease, retains physiological functions independent of amyloid production that remain poorly understood. Here, we identify a conserved, non-amyloidogenic role for APP in regulating apoptotic cell degradation via the endolysosomal pathway. Using Drosophila APPL as a model, structure-function analysis demonstrated that the intracellular internalization domain of APPL, but not its secreted ectodomain, is required for efficient apoptotic cell degradation. Immunoprecipitation coupled with mass spectrometry revealed a physical interaction between APPL and the microtubule severing ATPase Spastin, mediated by the microtubule-interacting and trafficking domain of Spastin. APPL interacts with Spastin on endosomal microtubules and modulates the dynamics of the Spastin-ESCRT-III complex, enabling Spastin to sever microtubules and promote endosomal tubule fission. Loss of APPL disrupts this process, causing aberrant endosomal tubulation and impaired lysosome biogenesis. Furthermore, it compromises the function of residual lysosomes, characterized by reduced acidity, diminished proteolytic activity, and increased lysosomal damage, which ultimately impairs the degradation of engulfed apoptotic cells. Critically, this phenotype is evolutionarily conserved in C. elegans and mice. Together, these findings establish a conserved APP-Spastin axis that regulates endolysosomal homeostasis and apoptotic cargo digestion. This reveals a critical non-amyloidogenic function of APP in maintaining tissue homeostasis through efficient efferocytosis, with broad implications for inflammatory and neurodegenerative disorders that warrant further investigation.

The endoplasmic reticulum (ER) and its muscle-specialized form, the sarcoplasmic reticulum (SR), are crucial organelles in muscle cells, involved notably in protein synthesis, calcium regulation and muscle contraction. A well-known process involved in ER remodeling and homeostasis is ER-phagy, also called reticulophagy, a selective form of autophagic process in which ER-phagy receptors mediate the delivery of ER portions to lysosomes for degradation. SH3KBP1 is an adaptor protein involved in membrane trafficking. Recently, it was shown to control ER morphology and SR formation in striated skeletal muscle. In this study, we demonstrate that SH3KBP1 can bind to LC3B and CKAP4 proteins, bridging ER to autophagosome membranes, and is degraded by autophagy, in developing muscle fibers. Moreover, SH3KBP1 down-regulation impacts basal autophagy efficiency and ER-phagy stimulation; it also impairs the turnover of numerous ER-resident proteins. Our work highlights a new role for SH3KBP1 as a soluble ER-phagy receptor in striated skeletal muscle.

PurposeMild therapeutic hypothermia (MTH) preserves cochlear function in animal models and is now entering early-phase human trials for hearing preservation. However, the extent to which the human cochlea can actually be cooled, and the mechanisms underlying MTH, remain unclear, in part because blood perfusion is expected to oppose localized cooling. In this study we evaluated the impact of blood flow on human cochlear temperature exposed to the MTH device using a combined experimental and computational approach. MethodsTemperature measurements were obtained from a human cadaver skull exposed to a commercial MTH device. These data were used to validate a three-dimensional bioheat transfer model incorporating realistic skull anatomy. The validated model was subsequently extended to include physiological blood perfusion in the internal carotid artery; a major heat source located near the cochlea. Finally, the in silico model was further expanded to incorporate the surrounding skin and brain tissues. ResultsIncorporating blood flow in internal carotid artery substantially altered predicted cochlear temperature distributions, highlighting the importance of localized vascular heat transport in the human cochlea during MTH. Although cochlear cooling was attenuated in the presence of perfusion, the therapeutic effects of MTH may not depend solely on the magnitude of local intracochlear temperature reduction. Additional mechanisms, such as reduced facial surface temperature, may also contribute to its efficacy. ConclusionThe validated in silico model provides a physiologically realistic framework for evaluating human cochlear thermal responses, investigating MTH mechanisms, and optimizing temperature-based strategies for hearing preservation.

WNT11, a non-canonical WNT ligand, plays well-defined roles in development and tissue architecture; however, its function in cancer remains ambiguous. Here, we characterize WNT11 as a context-dependent suppressor of cancer stemness, invasion, and in vivo tumor formation in human epithelial cancer models. We show that WNT11 upregulation reduces the expression of stemness-promoting genes, suppresses epithelial-to-mesenchymal transition, and inhibits sphere formation and tumor growth. Conversely, downregulation of WNT11 enhances these aggressive malignant properties of cancer cells. Mechanistically, we found that the ability of WNT11 to inhibit RAC1 GTPase activation is essential for its regulation of invasion and self-renewal. In cells unresponsive to WNT11, the connectivity between WNT11 and RAC1 activity is disengaged. Direct manipulation of RAC1 activity in these cells recapitulates the phenotype and molecular signature of WNT11-responsive cells, establishing RAC1 as a critical effector of WNT11-mediated tumor suppression. Taken together, these findings identify the cellular context in which WNT11 suppresses RAC1 activation as a key determinant of its anti-tumor effects and provide a mechanistic framework for understanding the diverse, and sometimes opposing, roles of WNT11 reported in cancer.

BackgroundPalliative radiation therapy (RT) for metastatic spine disease significantly increases the risk of vertebral fractures. However, the temporal mechanisms underlying radiation-induced vertebral bone fragility remain poorly understood. ObjectiveTo evaluate the longitudinal effects of a single high-dose irradiation, simulating palliative RT, on vertebral bone mechanical, architectural, and compositional properties in a healthy, skeletally mature rat model. MethodsThirty-one male Sprague Dawley rats received a single 15 Gy lumbar spine irradiation (IR). L4 vertebrae were assessed across all groups (irradiation: 7, 14, and 28 days post-IR, controls: at 0 and 28 days post-IR) for compressive strength and stiffness, micro-CT-derived bone composition and trabecular indices, serum bone turnover markers (NTX and BAP) and advanced glycation endproducts (AGEs). ResultsIrradiation induced progressive deterioration of vertebral bone mechanical properties, with strength decreasing up to 44% and stiffness up to 38% by 28 days post-IR, compared to 0- day controls. Trabecular bone exhibited reduced BMD, BV/TV, and Tb.N with increased Tb.Sp, a shift toward a more rod-like structure. Early post-IR changes suggested disrupted bone remodeling, characterized by elevated NTX and AGEs, but decreased BAP. Multivariable regression demonstrated that Tb.Th and AGEs were independent predictors of stiffness, collectively explaining 61% of its variance. DiscussionHigh-dose irradiation induces sustained temporal degradation of vertebral mechanical properties driven by both trabecular architectural deterioration and alterations in bone matrix quality. Measures of bone composition and non-enzymatic bone turnover suggest this early damage was driven by disruption of bone cellular homeostasis, favoring increased resorption over formation. These findings support that radiation impairs both structural integrity and pre-yield mechanical behavior, providing mechanistic insight into the elevated fracture risk observed clinically after irradiation for metastatic spine disease. Lay summaryThis study used a rat model to mimic palliative radiation therapy for cancer that has spread to the spine and evaluated the changes in bone quality up to 28 days post-therapy. We found that irradiation progressively weakened the structural integrity and composition of the bones in the spine and disrupted the normal balance of bone breakdown and repair, leading to greater bone loss and fragility. Our findings provide insight into the increased risk of fractures observed in patients receiving radiation therapy to the spine and may support efforts to better protect bone health during treatment.

Cooling the skin may increase its tolerance to mechanical loading and decrease the risk of developing pressure ulcers. Yet, the mechanisms of action (e.g. cooling-modulation of cytotoxic, post-occlusive hyperaemia), and their individual variability, remain unclear. We investigated the effects of different cooling levels (24{degrees}C and 16{degrees}C) on microvascular, inflammatory and perceptual responses to mechanical loading of the sacrum in healthy young (N=23) and older adults (N=19), and in spinal cord injury patients (SCI; N=10). Healthy participants underwent 45-min loading (~60 mmHg) and 20-min unloading of the sacrum, using an instrumented indenter probe set at either 38{degrees}C (control condition), 24{degrees}C or 16{degrees}C. SCI participants completed a more conservative protocol (i.e. 25min, ~45mmHg loading, 38{degrees}C and 16{degrees}C conditions). Pre-insult skin structure was characterised with optical coherence tomography; skin blood flow (SkBF) at the loading site was continuously measured, alongside thermal acceptability; and post-insult inflammatory responses were determined via skin-sebum cytokines analyses. Compared to control, 24{degrees}C- and 16{degrees}C-cooling induced a similar ~8-fold decrease in peak post-occlusive reactive hyperaemia in healthy participants, with similar temperature-related differences observed in SCI. Pro-inflammatory cytokines decreased post-insult; yet this occurred similarly across all temperatures and groups. The majority of participants ([&ge;]70%) rated both 24{degrees}C- and 16{degrees}C-cooling as thermally acceptable. We conclude that cooling is a potent modulator of the skin microvascular response to mechanical loading in younger, older, and vulnerable skin (SCI). These findings can inform design parameters for thermal technology aimed at preventing the loss of skin integrity (e.g. integrating 24{degrees}C-cooling in support surfaces and skin wearables).

The main olfactory epithelium (MOE) is the primary site of olfaction and consists of multiple cell types including olfactory sensory neurons (OSNs), sustentacular cells, and immune cells. Neuroimmune interactions in epithelial tissues are critical in maintaining tissue function, but how OSNs and immune cells interact in the MOE in healthy and diseased states is largely unknown. Cellular responses in the MOE determine how and whether OSNs maintain olfactory function and are repaired or replenished following inflammatory environmental exposures. We hypothesized that acute nasal aeroallergen exposure alters immune cell function in the MOE to elicit a neuroprotective response, thereby preserving OSN function. We developed an environmental aeroallergen exposure consisting of one week of daily intranasal house dust mite extract (HDM) instillations. Spectral flow cytometry indicated only subtle changes in resident immune cells proportions and phenotypes in the MOE. Immunohistochemical evaluation did not reveal extensive changes in immune cell distribution in the sensory epithelium or lamina propria, but instead we observed increases in axonal olfactory marker protein (OMP) expression in the lamina propria, where resident immune cells are most abundant. To evaluate the effects of HDM exposure on OSN function, we performed live ex vivo Ca2+ imaging of MOEs from HDM- and sham-exposed transgenic mice using objective-coupled planar illumination (OCPI) microscopy. OSN responses to multiple odorants revealed increased chemosensory sensitivity and decreased across-trial adaptation in HDM-treated epithelia. These results indicate that short-term nasal aeroallergen exposure minimally alters immune cell phenotypes, and instead induces functional changes in OSN physiology that preserve olfactory function.

1. Sandy beaches along urbanized coasts are increasingly managed through beach nourishment and hard infrastructure, yet these interventions often constrain natural dune-building processes. Along the Belgian coast, where much of the beach-dune interface is bordered by dikes, promenades and intensive recreation, strandline vegetation may provide an overlooked mechanism for retaining sand and initiating embryo dune development. 2. We assessed the potential for four pioneer dune plant species (Cakile maritima, Calamagrostis arenaria, Elymus farctus and Salsola kali) to establish, develop vegetation cover and contribute to sand accumulation along the Belgian coast. Using field surveys from 2017-2023, LiDAR-derived beach elevation and annual sediment dynamics, we modelled species occurrence and abundance/cover in low-disturbance reference zones and projected these relationships across the wider coastline. 3. Occurrence models identified where abiotic conditions allow plants to establish and persist until the late growing season, whereas zero-inflated abundance/cover models estimated expected vegetation development across environmental gradients. Predicted occurrence was widespread for several species, suggesting that the abiotic gradients modelled here are not the primary constraints on potential establishment across large parts of the coast. In contrast, expected abundance/cover showed stronger species-specific responses, particularly to sand accretion, indicating that sediment dynamics mainly affect post-establishment vegetation development rather than occurrence alone. 4. Independent field measurements of embryo dunes showed positive relationships between vegetation cover and local sand accumulation for all four species. When scaled using spatial predictions of potential abundance/cover, pioneer vegetation could retain substantial volumes of sand, with Cakile maritima contributing the largest share, followed by Salsola kali, Elymus farctus and Calamagrostis arenaria. Estimated volumes depended on assumptions about whether vegetation occurs as dispersed units or aggregated patches. 5. Synthesis and applications. Our results show that, even along a heavily urbanized and nourished coastline, abiotic conditions can support strandline vegetation and embryo dune initiation where disturbance is reduced. Management actions such as limiting trampling, adapting beach cleaning and protecting strandline vegetation could enhance the retention of nourished sand and support nature-based coastal defense. Rather than replacing engineered interventions, strandline vegetation may increase the efficiency with which available sediment is retained within the beach-dune system.

Analyses of human breast carcinomas (BCs) and premalignant breast lesions show that the loss of RASSF1A is an early event in the development of ER+ BCs, which correlates linearly with malignant progression. This observation suggests that RASSF1A inhibition is important for the development and progression of ER+ BCs. In addition to RASSF1A, concurrent caveolin-1 (Cav-1) inhibition may further promote ER+ breast carcinogenesis. In the present study, transgenic Rassf1a-/- and Cav-1(-/-) single as well as Rassf1a-/-, Cav-1(-/-) double knockout mice were used to investigate the impact of single or combined Rassf1a and Cav-1 inactivation on BC initiation. Loss of either one or both proteins led to different, pre-malignant histopathological alterations within the mammary glands of the mice, but not to fully developed BC, confirming that Rassf1a and Cav-1 are both important for maintaining the integrity of mammary gland epithelial structure, but suggesting that further intracellular changes or extracellular factors are required for the development of luminal BC when both genes are lost.

Sakumo Lagoon, a small (1 km2) semi-open coastal lagoon in Ghana, lies between the cities of Accra and Tema. The lagoon and its surrounding wetland were designated a Ramsar Site in 1992, mainly because it served as a refuge for 66 local and migratory bird species. Its ecology, and the biology of its major fish species, notably the blackchin tilapia (Sarotherodon melanotheron) were thoroughly studied in 1971, when the lagoon was a diverse, mainly brackish ecosystem supporting a traditionally and well-managed fishery. In 2016-2017, another study found the lagoon mostly covered by floating vegetation and plastic waste. Finally, in 2024, a visual survey established that the floating vegetation had been almost completely replaced by terrestrial plants, with only a few square meters of garbage-strewn water in front of a culvert connecting the lagoon to the open sea. Several lagoons along the coast of Ghana have been similarly lost to urban sprawl and its various forms of pollution, but Sakumo Lagoon is a Ramsar Site, and its imminent disappearance should not remain undocumented.

Abstract Introduction: Proximal humerus fractures account for approximately 5-6% of all adult fractures and are primarily managed nonoperatively. Healing is conventionally monitored with radiographs, with radiopaque callus formation indicating healing. Visible radiographic callus appears weeks after biological union begins. Ultrasound provides a dynamic, radiation-free, and cost-effective method that can detect early callus formation before x-ray visibility. Although ultrasound has demonstrated utility for fracture healing in the clavicle and humeral shaft, its role in proximal humerus fractures remains unclear. Methods: This single-centre prospective study will be conducted in two phases. The pilot phase will measure inter-rater reliability for ultrasound detection of early callus formation at 2 and 4 weeks post-injury. Ten patients with proximal humerus fractures treated nonoperatively will undergo standardized anterior and lateral scans. Each patient will generate four saved images (short- and long-axis views), producing forty anonymized images independently reviewed by two raters. The prospective cohort phase will recruit approximately thirty additional patients. Results: Reliability will be quantified using Cohens kappa. A power calculation will be performed after pilot analysis. Results from the prospective cohort phase will help determine the association and predictive value of early ultrasound-detected bridging callus for radiographic and clinical union at three and six months. Patient reported outcome measures will be assessed using the Quick Disabilities of Arm, Shoulder and Hand (QuickDASH) questionnaire. Discussion: This study will develop and validate a standardized ultrasound protocol for assessing early fracture healing in proximal humerus fractures. By establishing both inter-rater reliability and predictive value, the findings may support ultrasound as a reproducible, radiation-free adjunct to conventional imaging and enable earlier identification of union status.