Immune-regulated IDO1-dependent tryptophan metabolism is source of one-carbon units for pancreatic cancer and stellate cells.

Cancer cells adapt their metabolism to support elevated energetic and anabolic demands of proliferation. Folate-dependent one-carbon metabolism is a critical metabolic process underpinning cellular proliferation supplying carbons for the synthesis of nucleotides incorporated into DNA and RNA. Recent research has focused on the nutrients that supply one-carbons to the folate cycle, particularly serine. Tryptophan is a theoretical source of one-carbon units through metabolism by IDO1, an enzyme intensively investigated in the context of tumor immune evasion. Using in vitro and in vivo pancreatic cancer models, we show that IDO1 expression is highly context dependent, influenced by attachment-independent growth and the canonical activator IFNγ. In IDO1-expressing cancer cells, tryptophan is a bona fide one-carbon donor for purine nucleotide synthesis in vitro and in vivo. Furthermore, we show that cancer cells release tryptophan-derived formate, which can be used by pancreatic stellate cells to support purine nucleotide synthesis.

The Cytokine TNF Promotes Transcription Factor SREBP Activity and Binding to Inflammatory Genes to Activate Macrophages and Limit Tissue Repair.

Cytokine tumor necrosis factor (TNF)-mediated macrophage polarization is important for inflammatory disease pathogenesis, but the mechanisms regulating polarization are not clear. We performed transcriptomic and epigenomic analysis of the TNF response in primary human macrophages and revealed late-phase activation of SREBP2, the master regulator of cholesterol biosynthesis genes. TNF stimulation extended the genomic profile of SREBP2 occupancy to include binding to and activation of inflammatory and interferon response genes independently of its functions in sterol metabolism. Genetic ablation of SREBP function shifted the balance of macrophage polarization from an inflammatory to a reparative phenotype in peritonitis and skin wound healing models. Genetic ablation of SREBP activity in myeloid cells or topical pharmacological inhibition of SREBP improved skin wound healing under homeostatic and chronic inflammatory conditions. Our results identify a function and mechanism of action for SREBPs in augmenting TNF-induced macrophage activation and inflammation and open therapeutic avenues for promoting wound repair.

Engineering aquatic plant community composition on floating treatment wetlands can increase ecosystem multifunctionality.

Phytoremediation using floating treatment wetlands (FTWs) is an emerging nature-based solution for freshwater restoration. However, the potential to design these systems by manipulating macrophyte community composition to provide multiple ecosystem services remains unexplored. Using a tank experiment, we simulated aquatic environments impacted by multiple pollutants and employed a comparative ecological approach to design emergent macrophyte communities using the trait of plant stature (plant height) to structure communities. Ecosystem functions were quantified, and a threshold-based method used to compute an ecosystem multifunctionality index that was weighted based on three different management-driven restoration objectives: equal importance, phytoremediation, and regulation and cultural services. Across all restoration scenarios, ecosystem multifunctionality was higher when community types performed more diverse functions. Small emergent plant communities outperformed all other community types due to their increased provision of both regulation and maintenance, cultural, and provisioning services. Conversely, large emergent communities that are more typical candidates for phytoremediation had the highest levels of multifunctionality only when function was lower. Arranging emergent macrophytes in mixed-statured communities led to intermediate or poorer performance both in terms of multifunctionality and specific functions, suggesting that diversity on the plant stature axis leads to negative plant interactions and represents a 'worst of both worlds' combination. Employing comparative ecology to generalise plant selection by stature demonstrates that large emergent macrophytes are more likely to better deliver provision-based services, while small emergent communities can provide additional benefits from cultural and regulatory services. Selecting macrophytes for FTWs employed in freshwater restoration by stature is a simple and widely applicable approach for designing plant communities with predictable outcomes in terms of (multiple) ecosystem service provision and highlights the need for environmental managers to closely align restoration objectives with potential community types.

Multi-pollutant removal dynamics by aquatic plants in monoculture or mixed communities.

Much of our knowledge about the phytoremediation potential of floating treatment wetlands (FTWs) comes from studies focusing on the removal of single pollutants, often by a single plant species. Here, we quantify the potential of FTWs planted with varying proportions of the emergent monocots Typha latifolia, Glyceria maxima, and Phragmites australis to simultaneously remove a suite of eleven nutrient/metalloid pollutants. Pollutants most readily removed from water included total inorganic nitrogen (TIN), K and Mn, whilst P, Zn and Cu showed a moderate removal efficiency, and Mg, Ca, Na, Cr, and Fe were poorly removed. Root length within a FTW was correlated with lower concentrations of remaining in the water, whilst plant uptake and tissue sequestration was more important for reducing concentrations of Mn, TIN, P, and Fe. The effect of community composition over time was greatest for the removal of Zn, with FTWs containing T. latifolia having the strongest effect; community type was less important for the removal of TIN, Mg, K, and Na. Plant tissue sequestration was important for reducing concentrations of Mn, TIN, P and Fe in the water, with median uptake values all greater than 12.5%. Importantly, the removal of some pollutants (e.g., Cu) increased with retention time. Therefore, depending on the management objective, FTWs generally perform better where and when residence times are longer e.g., in ponds or streams under low flow, and assembling FTW communities with varying traits and associated removal mechanisms can allow several pollutants to be remediated at once.

Human lesions and animal studies link the claustrum to perception, salience, sleep and pain.

The claustrum is the most densely interconnected region in the human brain. Despite the accumulating data from clinical and experimental studies, the functional role of the claustrum remains unknown. Here, we systematically review claustrum lesion studies and discuss their functional implications. Claustral lesions are associated with an array of signs and symptoms, including changes in cognitive, perceptual and motor abilities; electrical activity; mental state; and sleep. The wide range of symptoms observed following claustral lesions do not provide compelling evidence to support prominent current theories of claustrum function such as multisensory integration or salience computation. Conversely, the lesions studies support the hypothesis that the claustrum regulates cortical excitability. We argue that the claustrum is connected to, or part of, multiple brain networks that perform both fundamental and higher cognitive functions. As a multifunctional node in numerous networks, this may explain the manifold effects of claustrum damage on brain and behaviour.

Methods for informing people with amyotrophic lateral sclerosis/motor neuron disease of their diagnosis.

BACKGROUND: Amyotrophic lateral sclerosis (ALS), also known as motor neuron disease (MND), causes increasing physical impairment and disability. People with ALS/MND face huge physical challenges, and the diagnosis can be a source of great psychological distress for both people with ALS/MND and their carers. In such a context, how news of the diagnosis is broken is important. At present, there are no systematic reviews of methods for informing people with ALS/MND of their diagnosis. OBJECTIVES: To examine the effects and effectiveness of different methods for informing people of a diagnosis of amyotrophic lateral sclerosis/motor neuron disease (ALS/MND), including effects on the person's knowledge and understanding of their disease, its treatment, and care; and on coping and adjustment to the effects of ALS/MND, its treatment, and care. SEARCH METHODS: We searched the Neuromuscular Specialised Register, CENTRAL, MEDLINE, Embase, PsycINFO, and two trials registers (February 2022). We contacted individuals or organisations to locate studies. We contacted study authors to obtain additional unpublished data. SELECTION CRITERIA: We planned to include randomised controlled trials (RCTs) and quasi-RCTs of techniques for informing people with ALS/MND of their diagnosis. We planned to include adults (aged 17 years or over) with ALS/MND, according to the El Escorial criteria. DATA COLLECTION AND ANALYSIS: Three review authors independently reviewed the results of the search to identify RCTs, and three review authors identified non-randomised studies to include in the discussion section. We planned that two review authors would independently extract data, and three would assess the risk of bias in any included trials. MAIN RESULTS: We did not identify any RCTs that met our inclusion criteria. AUTHORS' CONCLUSIONS: There are no RCTs that evaluate different communication strategies for breaking the bad news for people diagnosed with ALS/MND. Focused research studies are needed to assess the effectiveness and efficacy of different communication methods.

Response to "The opinion of French pulmonologists and palliative care physicians on non-invasive ventilation during palliative sedation at end of life: a nationwide survey''.

We have several major concerns about this article [Guastella et al 2021]. Although it states that it is about palliative sedation, it is not. Rather, it is about the French Claeys Leonetti Law about Continuous Deep Sedation (CDS) at the end of life [Loi n°2016-87].

Quality of life measurement tools for people with dementia and intellectual disabilities: A systematic review.

BACKGROUND: Adults with intellectual disabilities are an at-risk group of developing dementia. In the absence of a cure for dementia, emphasis on treatment is the promotion of Quality of life (QoL). The aim of this review is to identify and describe QoL tools for people with intellectual disabilities and dementia. METHOD: A systematic review was carried out using 10 databases and papers from up to March year 2021. RESULTS: Two instruments were identified and examined. The QoL in late-stage dementia, which showed evidence of good levels of internal consistency, intra-rater reliability, test-retest reliability, and convergent validity. The Dementia Quality of Life - proxy was also used; however, its psychometric properties have yet to be studied within the intellectual disabilities population. CONCLUSION: It is recommended instruments should be developed and psychometrically tested specifically for adults with intellectual disabilities and dementia to help inform policy makers, measure outcomes of interventions and personal outcomes.

An integrated, modular approach to data science education in microbiology.

We live in an increasingly data-driven world, where high-throughput sequencing and mass spectrometry platforms are transforming biology into an information science. This has shifted major challenges in biological research from data generation and processing to interpretation and knowledge translation. However, postsecondary training in bioinformatics, or more generally data science for life scientists, lags behind current demand. In particular, development of accessible, undergraduate data science curricula has the potential to improve research and learning outcomes as well as better prepare students in the life sciences to thrive in public and private sector careers. Here, we describe the Experiential Data science for Undergraduate Cross-Disciplinary Education (EDUCE) initiative, which aims to progressively build data science competency across several years of integrated practice. Through EDUCE, students complete data science modules integrated into required and elective courses augmented with coordinated cocurricular activities. The EDUCE initiative draws on a community of practice consisting of teaching assistants (TAs), postdocs, instructors, and research faculty from multiple disciplines to overcome several reported barriers to data science for life scientists, including instructor capacity, student prior knowledge, and relevance to discipline-specific problems. Preliminary survey results indicate that even a single module improves student self-reported interest and/or experience in bioinformatics and computer science. Thus, EDUCE provides a flexible and extensible active learning framework for integration of data science curriculum into undergraduate courses and programs across the life sciences.

Decoding river pollution trends and their landscape determinants in an ecologically fragile karst basin using a machine learning model.

Karst watersheds accommodate high landscape complexity and are influenced by both human-induced and natural activity, which affects the formation and process of runoff, sediment connectivity and contaminant transport and alters natural hydrological and nutrient cycling. However, physical monitoring stations are costly and labor-intensive, which has confined the assessment of water quality impairments on spatial scale. The geographical characteristics of catchments are potential influencing factors of water quality, often overlooked in previous studies of highly heterogeneous karst landscape. To solve this problem, we developed a machining learning method and applied Extreme Gradient Boosting (XGBoost) to predict the spatial distribution of water quality in the world's most ecologically fragile karst watershed. We used the Shapley Addition interpretation (SHAP) to explain the potential determinants. Before this process, we first used the water quality damage index (WQI-DET) to evaluate the water quality impairment status and determined that CODMn, TN and TP were causing river water quality impairments in the WRB. Second, we selected 46 watershed features based on the three key processes (sources-mobilization-transport) which affect the temporal and spatial variation of river pollutants to predict water quality in unmonitored reaches and decipher the potential determinants of river impairments. The predicting range of CODMn spanned from 1.39 mg/L to 17.40 mg/L. The predictions of TP and TN ranged from 0.02 to 1.31 mg/L and 0.25-5.72 mg/L, respectively. In general, the XGBoost model performs well in predicting the concentration of water quality in the WRB. SHAP explained that pollutant levels may be driven by three factors: anthropogenic sources (agricultural pollution inputs), fragile soils (low organic carbon content and high soil permeability to water flow), and pollutant transport mechanisms (TWI, carbonate rocks). Our study provides key data to support decision-making for water quality restoration projects in the WRB and information to help bridge the science:policy gap.

Validation of a sustainable internationally monitored cervical cancer screening system using a visual smartphone inspection in Kinshasa, Democratic Republic of Congo.

OBJECTIVE: To determine the sensitivity, specificity, and positive and negative predictive values of a cervical cancer screening program based on visual inspection with acetic acid and Lugol's iodine using a smartphone in a sub-urban area of very low resources in Kinshasa (Democratic Republic of Congo). METHODS: This cross-sectional validation study was conducted at Monkole Hospital and it included women between the ages of 25-70 years after announcing a free cervical cancer screening campaign through posters placed in the region of our hospital. Questionnaires collected sociodemographic and behavioral patients characteristics. In the first consultation, we gathered liquid-based cytology samples from every woman. At that time, local health providers performed two combined visual inspection techniques (5% acetic acid and Lugol's iodine) while a photograph was taken with a smartphone. Two international specialists evaluated the results of the smartphone cervicography. When a visual inspection was considered suspicious, patients were offered immediate cryotherapy. Cytological samples were sent to the Pathology Department of the University of Navarra for cytological assessment and human papillomavirus (HPV) DNA genotyping. RESULTS: A total of 480 women participated in the study. The mean age was 44.6 years (range 25-65). Of all the patients, only 18.7% were infected with HPV (75% had high-risk genotypes). The most frequent high-risk genotype found was 16 (12.2%). The majority (88%) of women had normal cytology. After comparing combined visual inspection results with cytology, we found a sensitivity of 66.0%, a specificity of 87.8%, a positive predictive value of 40.7%, and a negative predictive value of 95.3% for any cytological lesion. The negative predictive value for high-grade lesions was 99.7%. CONCLUSIONS: Cervical cancer screening through combined visual inspection, conducted by non-specialized personnel and monitored by experts through smartphones, shows encouraging results, ruling out high-grade cytological lesions in most cases. This combined visual inspection test is a valid and affordable method for screening programs in low-income areas.

Paid carers' understanding and experiences of meaningful involvement in bereavement for people with intellectual disability when a significant other is dying.

BACKGROUND: Wider communication about death and dying for those with intellectual disabilities has been highlighted as being of key importance. OBJECTIVE: To gain the perspective of paid carers based in residential homes about meaningfully supporting individuals with intellectual disabilities in the bereavement process. METHODS: Semi-structured interviews were conducted with paid carers working across two residential homes. RESULTS: Four themes were identified: (a) challenges in having conversations about death and dying; (b) meaningful involvement of residents in the bereavement process; (c) the relationship between paid carers and residents (including the impact of a resident's death on paid carers); and (d) the support needs of paid carers. CONCLUSIONS: The recommendations from previous research of meaningfully involving people with intellectual disabilities in the bereavement process have not filtered fully into practice. Paid carers still highlight the need for specialist support and advice to help them discuss and increase involvement in death and dying.

End-of-life care of people with long-term neurological conditions.

Guidance and protocols of end-of-life care have been directed towards the care of patients with cancer. It is possible to extrapolate some of these to people with long-term neurological conditions, but there are obvious differences. Neurological conditions have widely different time courses of progression, making the timing of these discussions challenging. The common issues around end-of-life care include knowing when to start discussions, approaching advance planning, managing common symptoms, diagnosing the dying phase, withdrawing life-sustaining treatments, providing support for family and carers and judging how to involve specialist palliative care teams. End-of-life care needs close collaboration between neurology, specialist palliative care and general practice.

Survival of human enteric and respiratory viruses on plastics in soil, freshwater, and marine environments.

The public health significance of plastics and microplastics in different environmental matrices has mainly focused on the toxicological effects of human ingestion. But these pollutants can also harbour pathogenic bacteria as the surfaces of plastics in the environment quickly become colonised by microbial biofilm. This novel microbial habitat has been termed the 'plastisphere' and could facilitate the survival and dissemination of important bacterial and fungal pathogens. Importantly, however, the role of plastic pollution as a secondary pathway for the transmission of human pathogenic viruses has never been addressed. Due to the high prevalence of both enteric and respiratory viruses in the population and in the environment, there is significant potential for human viruses to become associated with the plastisphere. In this review we critically evaluate current knowledge on the interaction of human enteric and respiratory viruses with plastic surfaces and identify the main environmental conditions and plastic characteristics that could affect virus survival and persistence in the environment. Our hypothesis is that the plastisphere can enhance the adhesion, survival and dissemination of human pathogenic viruses and potentially lead to more effective transfer and transmission of viral diseases within the environment. We identify key research questions needed to more fully assess the potential human health risks associated with viruses on plastic surfaces. These include understanding, (1) the mechanisms of viral attachment to either naked or biofilm-colonised plastic (2) how the structural characteristics of viruses (e.g., enveloped, or non-enveloped), affect their persistence in the plastisphere, (3) whether the plastisphere offers protection and increases the persistence of infectious viruses in soil, freshwater, and marine environments.

Single-cell transcriptomic analysis identifies extensive heterogeneity in the cellular composition of mouse Achilles tendons.

Tendon is a dense connective tissue that stores and transmits forces between muscles and bones. Cellular heterogeneity is increasingly recognized as an important factor in the biological basis of tissue homeostasis and disease, yet little is known about the diversity of cell types that populate tendon. To address this, we determined the heterogeneity of cell populations within mouse Achilles tendons using single-cell RNA sequencing. In assembling a transcriptomic atlas of Achilles tendons, we identified 11 distinct types of cells, including three previously undescribed populations of tendon fibroblasts. Prior studies have indicated that pericytes, which are found in the vasculature of tendons, could serve as a potential source of progenitor cells for adult tendon fibroblasts. Using trajectory inference analysis, we provide additional support for the notion that pericytes are likely to be at least one of the progenitor cell populations for the fibroblasts that compose adult tendons. We also modeled cell-cell interactions and identified previously undescribed ligand-receptor signaling interactions involved in tendon homeostasis. Our novel and interactive tendon atlas highlights previously underappreciated heterogeneity between and within tendon cell populations. The atlas also serves as a resource to further the understanding of tendon extracellular matrix assembly and maintenance and in the design of therapies for tendinopathies.

Widespread diversity in the transcriptomes of functionally divergent limb tendons.

KEY POINTS: Tendon is a hypocellular, matrix-rich tissue that has been excluded from comparative transcriptional atlases. These atlases have provided important knowledge about biological heterogeneity between tissues, and our study addresses this important gap. We performed measures on four of the most studied tendons, the Achilles, forepaw flexor, patellar and supraspinatus tendons of both mice and rats. These tendons are functionally distinct and are also among the most commonly injured, and therefore of important translational interest. Approximately one-third of the filtered transcriptome was differentially regulated between Achilles, forepaw flexor, patellar and supraspinatus tendons within either mice or rats. Nearly two-thirds of the transcripts that are expressed in anatomically similar tendons were different between mice and rats. The overall findings from this study identified that although tendons across the body share a common anatomical definition based on their physical location between skeletal muscle and bone, tendon is a surprisingly genetically heterogeneous tissue. ABSTRACT: Tendon is a functionally important connective tissue that transmits force between skeletal muscle and bone. Previous studies have evaluated the architectural designs and mechanical properties of different tendons throughout the body. However, less is known about the underlying transcriptional differences between tendons that may dictate their designs and properties. Therefore, our objective was to develop a comprehensive atlas of the transcriptome of limb tendons in adult mice and rats using systems biology techniques. We selected the Achilles, forepaw digit flexor, patellar, and supraspinatus tendons due to their divergent functions and high rates of injury and tendinopathies in patients. Using RNA sequencing data, we generated the Comparative Tendon Transcriptional Database (CTTDb) that identified substantial diversity in the transcriptomes of tendons both within and across species. Approximately 30% of filtered transcripts were differentially regulated between tendons of a given species, and nearly 60% of the filtered transcripts present in anatomically similar tendons were different between species. Many of the genes that differed between tendons and across species are important in tissue specification and limb morphogenesis, tendon cell biology and tenogenesis, growth factor signalling, and production and maintenance of the extracellular matrix. This study indicates that tendon is a surprisingly heterogenous tissue with substantial genetic variation based on anatomical location and species.

hnRNPs Interacting with mRNA Localization Motifs Define Axonal RNA Regulons.

mRNA translation in axons enables neurons to introduce new proteins at sites distant from their cell body. mRNA-protein interactions drive this post-transcriptional regulation, yet knowledge of RNA binding proteins (RBP) in axons is limited. Here we used proteomics to identify RBPs interacting with the axonal localizing motifs of Nrn1, Hmgb1, Actb, and Gap43 mRNAs, revealing many novel RBPs in axons. Interestingly, no RBP is shared between all four RNA motifs, suggesting graded and overlapping specificities of RBP-mRNA pairings. A systematic assessment of axonal mRNAs interacting with hnRNP H1, hnRNP F, and hnRNP K, proteins that bound with high specificity to Nrn1 and Hmgb1, revealed that axonal mRNAs segregate into axon growth-associated RNA regulons based on hnRNP interactions. Axotomy increases axonal transport of hnRNPs H1, F, and K, depletion of these hnRNPs decreases axon growth and reduces axonal mRNA levels and axonal protein synthesis. Thus, subcellular hnRNP-interacting RNA regulons support neuronal growth and regeneration.

The microbial safety of seaweed as a feed component for black soldier fly (Hermetia illucens) larvae.

Farmed insects can offer an environmentally sustainable aquafeed or livestock feed ingredient. The value of black soldier fly (Hermetia illucens) (BSF) larvae could be improved by enrichment in omega-3 through the dietary inclusion of seaweed. However, the industry practice of drying seaweed at low temperatures to retain nutritional properties may benefit the survival of human pathogenic bacteria, particularly if the seaweed has been harvested from contaminated water. Here we have demonstrated that E. coli and E. coli O157:H7 died-off in seaweed dried at 50 °C, although both were detected in the dried powder following 72 h storage. V. parahaemolyticus fell below the level of detection in stored seaweed after drying at ≥ 50 °C, but L. monocytogenes remained detectable, and continued to grow in seaweed dried at ≤60 °C. Therefore, drying seaweed at low temperatures risks pathogen carry-over into insects destined for animal feed. BSF larvae reared on an artificially contaminated seaweed-supplemented diet also became contaminated by all four bacteria present in the supplement. Water quality at seaweed harvesting sites, seaweed desiccation, and insect rearing practices, represent critical points where development of regulatory standards could achieve targeted control of pathogenic hazards.

CDK8/19 Mediator kinases potentiate induction of transcription by NFκB.

The nuclear factor-κB (NFκB) family of transcription factors has been implicated in inflammatory disorders, viral infections, and cancer. Most of the drugs that inhibit NFκB show significant side effects, possibly due to sustained NFκB suppression. Drugs affecting induced, but not basal, NFκB activity may have the potential to provide therapeutic benefit without associated toxicity. NFκB activation by stress-inducible cell cycle inhibitor p21 was shown to be mediated by a p21-stimulated transcription-regulating kinase CDK8. CDK8 and its paralog CDK19, associated with the transcriptional Mediator complex, act as coregulators of several transcription factors implicated in cancer; CDK8/19 inhibitors are entering clinical development. Here we show that CDK8/19 inhibition by different small-molecule kinase inhibitors or shRNAs suppresses the elongation of NFκB-induced transcription when such transcription is activated by p21-independent canonical inducers, such as TNFα. On NFκB activation, CDK8/19 are corecruited with NFκB to the promoters of the responsive genes. Inhibition of CDK8/19 kinase activity suppresses the RNA polymerase II C-terminal domain phosphorylation required for transcriptional elongation, in a gene-specific manner. Genes coregulated by CDK8/19 and NFκB include IL8, CXCL1, and CXCL2, which encode tumor-promoting proinflammatory cytokines. Although it suppressed newly induced NFκB-driven transcription, CDK8/19 inhibition in most cases had no effect on the basal expression of NFκB-regulated genes or promoters; the same selective regulation of newly induced transcription was observed with other transcription signals potentiated by CDK8/19. This selective role of CDK8/19 identifies these kinases as mediators of transcriptional reprogramming, a key aspect of development and differentiation as well as pathological processes.

The disparity between regulatory measurements of E. coli in public bathing waters and the public expectation of bathing water quality.

The main objectives of the European Union (EU) Bathing Water Directive (BWD) 2006/7/EC are to safeguard public health and protect designated aquatic environments from microbial pollution. The BWD is implemented through legislation by individual EU Member States and uses faecal indicator organisms (FIOs) as microbial pollution compliance parameters to determine season-end bathing water classifications (either 'Excellent', 'Good', 'Sufficient' or 'Poor'). These classifications are based on epidemiological studies that have linked human exposure to FIOs with the risk of contracting a gastrointestinal illness (GI). However, understanding public attitudes towards bathing water quality, together with perceptions of relative exposure risks, is often overlooked and yet critically important for informing environmental management decisions at the beach and ensuring effective risk communication. Therefore, this study aimed to determine the effectiveness of current regulatory strategies for informing beach users about bathing water quality, and to assess public understanding of the BWD classifications in terms of exposure risk and public health. Two UK designated bathing waters were selected as case studies, and questionnaires were deployed to beach-users. The bathing waters had different classification histories and both had electronic signage in operation for communicating daily water quality predictions. The majority of respondents did not recognise the standardised EU bathing water quality classification signs, and were unaware of information boards or the electronic signs predicting the water quality on that particular day. In general, respondents perceived the bathing water at their respective beach to be either 'good' or 'sufficient', which were also the lowest classifications of water quality they would be willing to accept for bathing. However, the lowest level of risk of contracting a gastrointestinal illness that respondents would be willing to accept suggested a significant misunderstanding of the BWD classification system, with the majority (91%) of respondents finding only a <1% risk level acceptable. The 'Good' classification is much less stringent in terms of likelihood of GI. This study has shown that the current public understanding of the BWD classifications in terms of exposure risk and public health is limited, and an investment in methods for disseminating information to the public is needed in order to allow beach-users to make more informed decisions about using bathing waters.