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Ribosome biogenesis lies at the nexus of various signaling pathways coordinating protein synthesis with cell growth and proliferation. This process is regulated by well-described transcriptional mechanisms, but a growing body of evidence indicates that other levels of regulation exist. Here we show that the Ras/mitogen-activated protein kinase (MAPK) pathway stimulates post-transcriptional stages of human ribosome synthesis. We identify RIOK2, a pre-40S particle assembly factor, as a new target of the MAPK-activated kinase RSK. RIOK2 phosphorylation by RSK stimulates cytoplasmic maturation of late pre-40S particles, which is required for optimal protein synthesis and cell proliferation. RIOK2 phosphorylation facilitates its release from pre-40S particles and its nuclear re-import, prior to completion of small ribosomal subunits. Our results bring a detailed mechanistic link between the Ras/MAPK pathway and the maturation of human pre-40S particles, which opens a hitherto poorly explored area of ribosome biogenesis.

Abstract Self/non‐self discrimination is a fundamental requirement of life. Endogenous peptides presented by major histocompatibility complex class I (MHC I) molecules represent the essence of self for CD8 T lymphocytes. These MHC I peptides (MIPs) are collectively referred to as the immunopeptidome. From a systems‐level perspective, very little is known about the origin, composition and plasticity of the immunopeptidome. Here, we show that the immunopeptidome, and therefore the nature of the immune self, is plastic and moulded by cellular metabolic activity. By using a quantitative high‐throughput mass spectrometry‐based approach, we found that altering cellular metabolism via the inhibition of the mammalian target of rapamycin results in dynamic changes in the cell surface MIPs landscape. Moreover, we provide systems‐level evidence that the immunopeptidome projects at the cell surface a representation of biochemical networks and metabolic events regulated at multiple levels inside the cell. Our findings open up new perspectives in systems immunology and predictive biology. Indeed, predicting variations in the immunopeptidome in response to cell‐intrinsic and ‐extrinsic factors could be relevant to the rational design of immunotherapeutic interventions.

Abstract The mammalian target of rapamycin (mTOR) is a central regulator of cell growth and proliferation. mTOR signaling is frequently dysregulated in oncogenic cells, and thus an attractive target for anticancer therapy. Using CellDesigner, a modeling support software for graphical notation, we present herein a comprehensive map of the mTOR signaling network, which includes 964 species connected by 777 reactions. The map complies with both the systems biology markup language (SBML) and graphical notation (SBGN) for computational analysis and graphical representation, respectively. As captured in the mTOR map, we review and discuss our current understanding of the mTOR signaling network and highlight the impact of mTOR feedback and crosstalk regulations on drug‐based cancer therapy. This map is available on the Payao platform, a Web 2.0 based community‐wide interactive process for creating more accurate and information‐rich databases. Thus, this comprehensive map of the mTOR network will serve as a tool to facilitate systems‐level study of up‐to‐date mTOR network components and signaling events toward the discovery of novel regulatory processes and therapeutic strategies for cancer.

Mitogen-activated protein kinase 3/1 (Mapk3/1) pathway is critical for LH signal transduction during ovulation. However, the mechanisms remain incompletely understood. We hypothesized that Mapk pathway regulates ovulation through transcriptional regulation of ovulatory genes. To test this hypothesis we used immature mice superovulated with equine and human chorionic gonadotropins (eCG and hCG) and PD0325901, to inhibit hCG-induced Mapk3/1 activity. Mice received either the inhibitor PD0325901 (25 μg/g, i.p.) or vehicle at 2h before hCG stimulation. Administration of the inhibitor abolished Mapk3/1 phosphorylation in granulosa cells. While vehicle-treated mice ovulated normally, there were no ovulations in inhibitor-treated mice. First, we analyzed gene expression in granulosa cells at 0h, 1h and 4h post-hCG. There was expected hCG-driven increase in mRNA abundance of many ovulation-related genes including Ptgs2 in vehicle-treated granulosa cells, but not (P

In patients with melanoma, increased RAS/mitogen-activated protein kinase (MAPK) pathway activity is known to drive chemotherapy resistance. Here, the authors identify CDK12 as a downstream effector of the RAS/MAPK pathway and therapeutic target which mediates chemotherapy resistance through increased expression of DNA repair associated genes.

Summary: Immunotherapy remains underexploited in acute myeloid leukemia (AML) compared to other hematological malignancies. Currently, gemtuzumab ozogamicin is the only therapeutic antibody approved for this disease. Here, to identify potential targets for immunotherapeutic intervention, we analyze the surface proteome of 100 genetically diverse primary human AML specimens for the identification of cell surface proteins and conduct single-cell transcriptome analyses on a subset of these specimens to assess antigen expression at the sub-population level. Through this comprehensive effort, we successfully identify numerous antigens and markers preferentially expressed by primitive AML cells. Many identified antigens are targeted by therapeutic antibodies currently under clinical evaluation for various cancer types, highlighting the potential therapeutic value of the approach. Importantly, this initiative uncovers AML heterogeneity at the surfaceome level, identifies several antigens and potential primitive cell markers characterizing AML subgroups, and positions immunotherapy as a promising approach to target AML subgroup specificities.

Abstract CDK12 is a transcriptional cyclin-dependent kinase (CDK) that interacts with cyclin K to regulate different aspects of gene expression. The CDK12-cyclin K complex phosphorylates several substrates, including RNA polymerase II (Pol II), and thereby regulates transcription elongation, RNA splicing, as well as cleavage and polyadenylation. Because of its implication in cancer, including breast cancer and melanoma, multiple pharmacological inhibitors of CDK12 have been identified to date, including THZ531 and SR-4835. While both CDK12 inhibitors affect Poll II phosphorylation, we found that SR-4835 uniquely promotes cyclin K degradation via the proteasome. Using loss-of-function genetic screening, we found that SR-4835 cytotoxicity depends on a functional CUL4-RBX1-DDB1 ubiquitin ligase complex. Consistent with this, we show that DDB1 is required for cyclin K degradation, and that SR-4835 promotes DDB1 interaction with the CDK12-cyclin K complex. Docking studies and structure-activity relationship analyses of SR-4835 revealed the importance of the benzimidazole side-chain in molecular glue activity. Together, our results indicate that SR-4835 acts as a molecular glue that recruits the CDK12-cyclin K complex to the CUL4-RBX1-DDB1 ubiquitin ligase complex to target cyclin K for degradation.

Abstract How the genetic landscape governs a tumor’s response to immunotherapy remains poorly understood. To assess the immune-modulatory capabilities of 573 genes associated with altered cytotoxicity in human cancers, here we perform CRISPR/Cas9 screens directly in mouse lung cancer models. We recover the known immune evasion factors Stat1 and Serpinb9 and identify the cancer testis antigen Adam2 as an immune modulator, whose expression is induced by KrasG12D and further elevated by immunotherapy. Using loss- and gain-of-function experiments, we show that ADAM2 functions as an oncogene by restraining interferon and TNF cytokine signaling causing reduced presentation of tumor-associated antigens. ADAM2 also restricts expression of the immune checkpoint inhibitors PDL1, LAG3, TIGIT and TIM3 in the tumor microenvironment, which might explain why ex vivo expanded and adoptively transferred cytotoxic T-cells show enhanced cytotoxic efficacy in ADAM2 overexpressing tumors. Together, direct in vivo CRISPR/Cas9 screens can uncover genetic alterations that control responses to immunotherapies.

Dyslipidemia and autophagy have been implicated in the pathogenesis of blinding neovascular age-related macular degeneration (NV-AMD). VLDL receptor (VLDLR), expressed in photoreceptors with a high metabolic rate, facilitates the uptake of triglyceride-derived fatty acids. Since fatty acid uptake is reduced in Vldlr–/– tissues, more remain in circulation, and the retina is fuel deficient, driving the formation in mice of neovascular lesions reminiscent of retinal angiomatous proliferation (RAP), a subtype of NV-AMD. Nutrient scarcity and energy failure are classically mitigated by increasing autophagy. We found that excess circulating lipids restrained retinal autophagy, which contributed to pathological angiogenesis in the Vldlr–/– RAP model. Triglyceride-derived fatty acid sensed by free fatty acid receptor 1 (FFAR1) restricted autophagy and oxidative metabolism in photoreceptors. FFAR1 suppressed transcription factor EB (TFEB), a master regulator of autophagy and lipid metabolism. Reduced TFEB, in turn, decreased sirtuin-3 expression and mitochondrial respiration. Metabolomic signatures of mouse RAP-like retinas were consistent with a role in promoting angiogenesis. This signature was also found in human NV-AMD vitreous. Restoring photoreceptor autophagy in Vldlr–/– retinas, either pharmacologically or by deleting Ffar1, enhanced metabolic efficiency and suppressed pathological angiogenesis. Dysregulated autophagy by circulating lipids might therefore contribute to the energy failure of photoreceptors driving neovascular eye diseases, and FFAR1 may be a target for intervention.

The oncogene KRAS is frequently mutated in cancer, including colorectal cancer. Here, using a cell-surface proteomics approach, KRAS-mutated colorectal cancer cells are shown to express high levels of the copper transporter ATP7A, which has an essential roles in cancer cell survival and proliferation.

Abstract: Acute megakaryoblastic leukemia (AMKL) represents ∼10% of pediatric acute myeloid leukemia cases and typically affects young children (

Directed motility of cell clusters requires coordination of protrusion formation at the front of leader cells and contractility at the rear. Here the authors show that the kinase Misshapen restricts protrusions to the leader cell and promotes contractile forces at the rear of the cluster.

KRAS-driven cancers acquire profound metabolic dependencies that are intimately linked to tumor growth. Our work revealed that colorectal cancers that harbor KRAS mutations are addicted to copper metabolism. This adaptation renders tumor cells critically dependent on the copper transporter ATP7A, which reveals copper metabolism as a promising therapeutic target for KRAS-driven colorectal cancers.

Summary: In human cells, the expression of ∼1,000 genes is modulated throughout the cell cycle. Although some of these genes are controlled by specific transcriptional programs, very little is known about their post-transcriptional regulation. Here, we analyze the expression signature associated with all 687 RNA-binding proteins (RBPs) and identify 39 that significantly correlate with cell cycle mRNAs. We find that NF45 and NF90 play essential roles in mitosis, and transcriptome analysis reveals that they are necessary for the expression of a subset of mitotic mRNAs. Using proteomics, we identify protein clusters associated with the NF45-NF90 complex, including components of Staufen-mediated mRNA decay (SMD). We show that depletion of SMD components increases the binding of mitotic mRNAs to the NF45-NF90 complex and rescues cells from mitotic defects. Together, our results indicate that the NF45-NF90 complex plays essential roles in mitosis by competing with the SMD machinery for a common set of mRNAs.

Summary: AXL is activated by its ligand GAS6 and is expressed in triple-negative breast cancer cells. In the current study, we report AXL expression in HER2-positive (HER2+) breast cancers where it correlates with poor patient survival. Using murine models of HER2+ breast cancer, Axl, but not its ligand Gas6, was found to be essential for metastasis. We determined that AXL is required for intravasation, extravasation, and growth at the metastatic site. We found that AXL is expressed in HER2+ cancers displaying epithelial-to-mesenchymal transition (EMT) signatures where it contributes to sustain EMT. Interfering with AXL in a patient-derived xenograft (PDX) impaired transforming growth factor β (TGF-β)-induced cell invasion. Last, pharmacological inhibition of AXL specifically decreased the metastatic burden of mice developing HER2+ breast cancer. Our data identify AXL as a potential anti-metastatic co-therapeutic target for the treatment of HER2+ breast cancers. : Metastasis is responsible for the majority of breast cancer deaths. Goyette et al. report that AXL is expressed in HER2+ human tumors that acquire aggressive features. Blockade of AXL in mice decreases metastasis. These results suggest that co-targeting AXL and HER2 may limit the metastatic progression of HER2+ breast cancer. Keywords: AXL, HER2, breast cancer, metastasis, EMT, PDX, AXL inhibitor

Beta-arrestins play central roles in the mechanisms regulating GPCR signalling and trafficking. Here the authors identify a selective inhibitor of the interaction between β-arrestin and the β2-adaptin subunit of the clathrin adaptor protein AP-2, which they use to dissect the role of the β-arrestin/β2-adaptin interaction in GPCR signalling.

Viticulture needs to satisfy consumers' demands for environmentally sound grape and wine production while envisaging adaptation options to diminish the impacts of projected climate change on future productivity. However, the impact of climate change and the adoption of adaptation levers on the environmental impacts of future viticulture have not been assessed. This study evaluates the environmental performance of grape production in two French vineyards, one located in the Loire Valley and another in Languedoc-Roussillon, under two climate change scenarios. First, the effect of climate-induced yield change on the environmental impacts of future viticulture was assessed based on grape yield and climate data sets. Second, besides the climate-induced yield change, this study accounted for the impacts of extreme weather events on grape yield and the implementation of adaptation levers based on the future probability and potential yield loss due to extreme events. The life cycle assessment (LCA) results associated with climate-induced yield change led to opposite conclusions for the two vineyards of the case study. While the carbon footprint of the vineyard from Languedoc-Roussillon is projected to increase by 29 % by the end of the century under the high emissions scenario (SSP5-8.5), the corresponding footprint is projected to decrease in the vineyard from the Loire Valley by approximately 10 %. However, when including the effect of extreme events and adaptation options, the life cycle environmental impacts of grape production are projected to drastically increase for both vineyards. For instance, under the SSP5-8.5 scenario, the carbon footprint for the vineyard of Languedoc-Roussillon is projected to increase fourfold compared to the current footprint, while it will rise threefold for the vineyard from the Loire Valley. The obtained LCA results emphasized the need to account for the impact of both climate change and extreme events on grape production under future climate change scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In the last decade, the exploration of deep space has become the objective of the national space programs of many countries. The International Space Exploration Coordination Group has set a roadmap whose long-range strategy envisions the expansion of human presence in the solar system to progress with exploration and knowledge and to accelerate innovation. Crewed missions to Mars could be envisaged by 2040. In this scenario, finding ways to use the local resources for the provision of food, construction materials, propellants, pharmaceuticals is needed. Plants are important resources for deep space manned missions because they produce phytochemicals of pharmaceutical relevance, are sources of food and provide oxygen which is crucial in bioregenerative life support systems. Growth analysis and plant biomass yield have been previously evaluated on Martian regolith simulants; however, molecular approaches employing gene expression analysis and proteomics are still missing. The present work aims at filling this gap by providing molecular data on a representative member of the Poaceae, Lolium multiflorum Lam., grown on potting soil and a Martian regolith simulant (MMS-1). The molecular data were complemented with optical microscopy of root/leaf tissues and physico-chemical analyses. The results show that the plants grew for 2 weeks on regolith simulants. The leaves were bent downwards and chlorotic, the roots developed a lacunar aerenchyma and small brownish deposits containing Fe were observed. Gene expression analysis and proteomics revealed changes in transcripts related to the phenylpropanoid pathway, stress response, primary metabolism and proteins involved in translation and DNA methylation. Additionally, the growth of plants slightly but significantly modified the pH of the regolith simulants. The results here presented constitute a useful resource to get a comprehensive understanding of the major factors impacting the growth of plants on MMS-1., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Separating raypaths in a multipath shallow-water environment is a challenge due to the interferences between them and the colored noise that exists in an ocean environment, especially for two raypaths that arrive close to each other. Thus, in this paper, a three-dimensional (3D) higher-order raypath separation in an array to array configuration is proposed. Performance tests using simulation data in a multipath environment, real data obtained in an ultrasonic waveguide, and ocean shallow-water data, respectively, illustrate that the proposed algorithm achieves a higher resolution and a stronger robustness compared to the existing algorithms.

We propose a modal approach developed in the framework of the paraxial approximation to investigate the effects of deterministic surface perturbations in a planar waveguide. In the first part, the sensitivity of the modal amplitudes is theoretically formulated for a three-dimensional perturbation at the air-water interface. When applied to a broadband ultrasonic signal in a laboratory tank experiment, this approach results in travel-time and amplitude fluctuations that are successfully compared to experimental data recorded between two vertical source-receiver arrays that span the ultrasonic waveguide. The nonlinear shape of the modal amplitude fluctuations is of particular interest and is due to the three-dimensional nature of the surface perturbation. In the second part, a time-harmonic inversion method is built in the paraxial single-scattering approximation to image the dynamic surface perturbation from the modal transmission matrix between two source-receiver arrays. Again, the inversion results for capillary-gravity surface perturbations are successfully compared to similar inversions performed from experimental data processed with a complete set of eigenbeams extracted between the two arrays.

A dramatic slowing down of acoustic wave transport in dense fish shoals is observed in open-sea fish cages. By employing a multi-beam ultrasonic antenna, we observe the coherent backscattering phenomenon. We extract key parameters of wave transport such as the transport mean free path and the energy transport velocity of diffusive waves from diffusion theory fits to the experimental data. The energy transport velocity is found to be about 10 times smaller than the speed of sound in water, a value that is exceptionally low compared with most observations in acoustics. By studying different models of the fish body, we explain the basic mechanism responsible for the observed very slow transport of ultrasonic waves in dense fish shoals. Our results show that, while the fish swim bladder plays an important role in wave scattering, other organs have to be considered to explain ultra-low energy transport velocities., (© 2021. The Author(s).)

Global value chains and climate change have a significant impact on water resources and increasingly threaten freshwater ecosystems. Recent methodological proposals for life cycle impact assessment (LCIA), evaluate water use impacts on freshwater habitats based on river hydraulic parameters alterations. However, they are limited to French rivers due to lack of global data and models. On this basis, this article proposes an approach to compute regionalized characterization factors for modeling river habitat change potential (HCP) induced by water consumption, potentially applicable worldwide. A simplified model is developed for fish guilds and invertebrates. Based on French datasets, it establishes a relationship between HCP and river hydraulic parameters. A methodology to derive discharge and hydraulic geometry at the reach scale is proposed and applied to European and Middle Eastern rivers below 60°N latitude. Regionalized HCPs are calculated at the river reach scale and aggregated at watershed. Then, the impact of agricultural water use in contrasted European and Middle Eastern countries is evaluated comparing the outcomes from the HCP and the Available Water Remaining (AWARE) models at the national scale, considering water supply mix data. The same analysis is carried out on selected river basins. Finally, result consistency, uncertainty and global applicability of the overall approach are discussed. The study demonstrates the reproducibility of the impact model developed for French rivers on any hydrographic network where comparable ecological, hydrological and hydraulic conditions are met. Furthermore, it highlights the need to characterize impacts at a higher spatial resolution in areas where HCP is higher. Large scale quantification of HCP opens the way to the operationalization of mechanistic LCIA models in which the habitat preferences of freshwater species are taken into account to assess the impacts of water consumption on biodiversity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Subglacial water flow strongly modulates glacier basal motion, which itself strongly influences the contributions of glaciers and ice sheets to sea level rise. However, our understanding of when and where subglacial water flow enhances or impedes glacier flow is limited due to the paucity of direct observations of subglacial drainage characteristics. Here, we demonstrate that dense seismic array observations combined with an innovative systematic seismic source location technique allows the retrieval of a two-dimensional map of a subglacial drainage system, as well as its day-to-day temporal evolution. We observe with unprecedented detail when and where subglacial water flows through a cavity-like system that enhances glacier flow versus when and where water mainly flows through a channel-like system that impedes glacier flow. Most importantly, we are able to identify regions of high hydraulic connectivity within and across the cavity and channel systems, which have been identified as having a major impact on the long-term glacier response to climate warming. Applying a similar seismic monitoring strategy in other glacier settings, including for ice sheets, may help to diagnose the susceptibility of their dynamics to increased meltwater input due to climate warming., Competing Interests: The authors declare no competing interest.

Decentralised wastewater management (DWM) systems are deployed in areas where the topography does not allow for gravity flow to a centralised system, or requires a complex and expensive pumping station network. Also, DWM systems are often the only option in rural areas where there are no sewage transport networks. This paper aims at addressing the question of the degree to which DWM systems can be considered as viable alternatives from an environmental point of view using the Life Cycle Assessment (LCA) methodology. First, the environmental sustainability is investigated to identify environmental hotspots in two (nature-based and engineered) onsite DWM systems. Second, DWM scenarios are compared against centralised wastewater management (CWM) scenarios using a whole-systems approach. Finally the boundary conditions under which a given DWM scenario performs better than a CWM scenario are discussed. Results show CWM scenarios were less sustainable than DWM scenarios on the resources endpoint due to their sewer infrastructures, however CWM scenarios performed better than DWM scenarios on the ecosystems quality endpoint due to their well-managed air emissions and discharges. While on human health no clear conclusion could be drawn. Finally, for relatively few households (subject of the study in rural areas) CWM scenarios did not score superior performances compared to DWM scenarios on all three endpoint indicators. Yet for a greater number of households it was impossible to decide in favour of decentralisation because of a lack of favourable consensus on all three endpoint indicators., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

An experimental comparison is reported here between two equivalent resonant subwavelength metasurfaces made of long aluminum beams glued closely together on a thin aluminum plate. One metasurface has a random distribution of the resonator beams, and the other has a regular square lattice of pitch 1.5 cm. The random lattice shows the "resonant" behavior of a typical metasurface, with a wide full bandgap for the first A 0 Lamb mode. Instead, the regular square lattice combines Fano resonance with Bragg scattering at the edges of the passband, thus creating anisotropy and a pseudo bandgap. Comparisons with numerical simulations are performed, with good agreement with the experimental data. The multimodal response of the beams is also responsible for double negativity in a narrow frequency band, and the event of a pseudo bandgap around this same flexural resonance. In addition, the scattering regimes for both the random and regular metasurfaces are characterized using coherent and incoherent signal analysis.

We study numerically the potential of a multimodal elastic metamaterial to filter and guide Lamb waves in a plate. Using a sub-wavelength array of elongated beams attached to the plate, and combining the coupling effects of the longitudinal and flexural motion of these resonators, we create narrow transmission bands at the flexural resonances of the beams inside the wide frequency bandgap induced by their longitudinal resonance. The diameter of the beams becomes the tuning parameter for selection of the flexural leakage frequency, without affecting the main bandgap. Finally, by combination of the monopolar and dipolar scattering effects associated with the coupled beam and plate system, we create a frequency-based multiplexer waveguide in a locally resonant metamaterial.

Ocean acoustic tomography is traditionally performed using the travel-time variations of an acoustic path between a source and a receiver. In the context of shallow-water tomography and multipath propagation, the different acoustic paths can be correctly identified if the source and the receiver are arrays of transducers. Here, a double-beamforming algorithm can be applied to extract a collection of eigenbeams from the raw acoustic dataset. In this study, four observables can be measured for each eigenbeam: the travel-time, the amplitude, and the emitting and receiving angles. In this study, the sensitivity kernel (SK) formulation is used to establish a quantitative relation between a perturbation of the surface of an ultrasonic waveguide and the emitting and receiving angles of each eigenbeam. This theoretical relation is experimentally demonstrated using a forward model experiment designed to measure the SK. The SK formulation is then used in a second experiment to quantitatively and dynamically image the propagation of a surface wave traveling across the surface of the waveguide. The inversion results show that the quality of the joint inversion of the emitting and receiving angles is higher than previous results based on amplitude or travel-time observables.

Life cycle assessment (LCA) has been widely applied in the wastewater industry, but inconsistencies in assumptions and methods have made it difficult for researchers and practitioners to synthesize results from across studies. This paper presents a critical review of published LCAs related to municipal wastewater management with a focus on developing systematic guidance for researchers and practitioners to conduct LCA studies to inform planning, design, and optimization of wastewater management and infrastructure (wastewater treatment plants, WWTPs; collection and reuse systems; related treatment technologies and policies), and to support the development of new technologies to advance treatment objectives and the sustainability of wastewater management. The paper guides the reader step by step through LCA methodology to make informed decisions on i) the definition of the goal and scope, ii) the selection of the functional unit and system boundaries, iii) the selection of variables to include and their sources to obtain inventories, iv) the selection of impact assessment methods, and v) the selection of an effective approach for data interpretation and communication to decision-makers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Multiple scattering of acoustic waves offers a noninvasive method for density estimation of a dense shoal of fish where traditional techniques such as echo-counting or echo-integration fail. Through acoustic experiments with a multi-beam sonar system in open sea cages, multiple scattering of sound in a fish shoal, and, in particular, the coherent backscattering effect, can be observed and interpreted quantitatively. Furthermore, a volumetric scan of the fish shoal allows isolation of a few individual fish from which target strength estimations are possible. The combination of those two methods allows for fish density estimation in the challenging case of dense shoals.