Pancreatic adenocarcinoma (PDAC) remains one of the most lethal malignancies worldwide, with limited improvements in long-term survival despite significant progress in systemic therapy. This review focuses predominantly on pancreatic ductal adenocarcinoma (PDAC), the most common and best-studied pancreatic malignancy, while referencing other pancreatic neoplasms only where directly relevant to intrapancreatic fat biology. Obesity has emerged as a major modifiable risk factor for PDAC. In addition, increasing attention has focused on ectopic fat depots, particularly intrapancreatic fat, as potential mediators linking metabolic disease to pancreatic carcinogenesis. This narrative review summarizes the classification and epidemiology of PDAC, the relationship between obesity and pancreatic cancer risk, the biology of intrapancreatic fat, as well as putative mechanisms by which intrapancreatic fat may promote pancreatic carcinogenesis. In addition, we will review current diagnostic approaches, therapeutic and preventive considerations, and key unanswered questions for future research on the relevance of intrapancreatic fat deposits. Collectively, available evidence supports intrapancreatic fat as a biologically plausible and potential mediator of pancreatic cancer risk, warranting further prospective investigation.

The coronavirus disease 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, has presented multifaceted health challenges. COVID-19 primarily targets the respiratory system but also affects multiple organ systems, including the endocrine system. Emerging evidence suggests interactions between thyroid function, the acute phase of COVID-19, and the prolonged symptoms known as post-COVID sequalae or long COVID. Several studies have reported that COVID-19 can induce thyroid dysfunction, leading to conditions such as thyroiditis and alterations in thyroid hormone levels. The mechanisms through which SARS-CoV-2 affects the thyroid include direct viral infection of thyroid cells, leading to viral thyroiditis, which causes inflammation and transient or sustained thyroid dysfunction, as well as an excessive systemic immune response (cytokine storm). This is associated with elevated levels of cytokines, such as IL-6, that disrupt thyroid function and lead to nonthyroidal illness syndrome (NTIS). Medications administered during the acute illness phase, such as corticosteroids and antiviral drugs, can also impact thyroid hormone actions. The involvement of the thyroid gland in long COVID, or postacute sequelae of SARS-CoV-2 infection, is an area not well defined, with potential implications for understanding and managing this condition. Persistent low-grade inflammation affecting thyroid function over time can lead to ongoing thyroiditis or exacerbate pre-existing thyroid conditions. Viral infections, including SARS-CoV-2, can trigger or worsen autoimmune thyroid diseases, such as Hashimoto's thyroiditis and Graves' disease. Long COVID may disrupt the hypothalamic-pituitary-adrenal (HPA) axis, which can, in turn, affect the hypothalamic-pituitary-thyroid (HPT) axis, leading to abnormal thyroid function. This review was designed to systematically capture recent literature on COVID-19-related thyroid dysfunction in the adult population, the prognostic consequences of thyroid dysfunction during COVID-19, and the effects of thyroid dysfunction on patients with long COVID. A comprehensive search of PubMed and EMBASE databases was conducted. The systematic review was performed based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Study quality was assessed using the Critical Appraisal Skills Programme (CASP). A total of 53 studies met the inclusion criteria. The review summarises recent findings and provides an update of the current understanding of thyroid dysfunction in COVID-19-related spectrum of disorders, underscoring the complex nature of SARS-CoV-2 infection and its far-reaching impacts on human health.

BACKGROUND:While coronavirus disease 2019 (COVID-19) is primarily a respiratory infection, few studies have characterised the immune response to COVID-19 in lung tissue. We sought to understand the pathogenic role of microenvironmental interactions and the extracellular matrix in post-mortem COVID-19 lung using an integrative multi-omic approach. METHODS:Post-mortem formalin-fixed paraffin-embedded lung tissue from fatal COVID-19 and nonrespiratory death control lung underwent multi-omic evaluation by Quantseq Bulk RNA sequencing, Nanostring GeoMx spatial transcriptomics, RNAscope, multiplex immunofluorescence and immunohistochemistry, to evaluate virus distribution, immune composition and the extracellular matrix. Markers of extracellular synthesis and breakdown were measured in the serum of 215 patients with COVID-19 and 54 healthy volunteer controls using ELISA. RESULTS:We found that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection was restricted to the pneumocytes and macrophages of early-stage disease. Spatial analyses revealed an immunosuppressive virus microenvironment, enriched for PDL1+IDO1+ macrophages and depleted of T-cells. Oligoclonal T-cells in COVID-19 lung showed no enrichment of SARS-CoV-2 specific T-cell receptors. Collagen VI was upregulated and contributed to alveolar wall thickening and impaired gas exchange in COVID-19 lung. Serum from COVID-19 patients showed increased levels of PRO-C6, a marker of collagen VI synthesis, predicted mortality in hospitalised patients. CONCLUSIONS:Our data refine the current model of respiratory COVID-19 with regard to virus distribution, immune niches and the role of the noncellular microenvironment in pathogenesis and risk stratification in COVID-19. We show that collagen deposition is an early event in the course of the disease.

Disclosure: E. Braybrook: None. K. Natasha: None. D. Grammatopoulos: None. Depressive symptoms experienced either during pregnancy or postpartum, collectively termed perinatal depression (PND) affects around 17% of women globally, with significant impact to both the mother and child’s health. The underpinning mechanisms are not yet fully understood, however dysregulation of the HPA axis is believed to be central, with impairment of neurotransmitter function also linked to inflammation. Additionally, the occurrence of depression antenatally is shown to be a significant risk factor in the development of postpartum depression. Clinicians currently use questionnaires as the primary tool to identify risk of depression, however their performance is inadequate and only one fifth of women who experience PND actively seek help. Biomarker-based screening strategies might offer an additional tool for earlier identification, stratification and development of targeted therapies. Plasma proteomics is emerging as a powerful tool, enabling both improved understanding of key biological processes through the generation of molecular profiles, and the identification of novel biomarkers for disease prediction. This study analysed serum samples from 260 women between 24-28 weeks gestation, with risk of depression assessed through the Edinburgh Postnatal Depression Questionnaire. To capture depressive symptoms either during pregnancy or postpartum, scores were obtained between 24-29 weeks gestation and again 6-10 weeks postpartum, with a cut-off score of 10 used to indicate increased risk. 92 inflammatory markers were analysed in the serum samples using Olink Proseek Multiplex Inflammation I panel, utilising a proximity extension assay. Differential expression analysis revealed distinct profiles between the antenatal and postnatal depression groups. Machine learning models (e.g. Random Forrest, Classification and Regression Tree and Pearsons Chi-square Statistic) were applied to the data using SPSS Modeler, with similarities across the outputs in key proteins identified (STAMPB, SIRT2, AXIN1, LAP TGF-beta-1, IL-10, MMP-10 and IL17C). In addition, differing psychosocial variables were highlighted as contributing factors across the two groups (history of anxiety or depression in antenatal and family history of PND in postnatal). Functional enrichment analyses further explored the biological functions of key proteins. This work highlights the value of targeted proteomics approaches coupled with machine learning in uncovering biomarker signatures that add to our understanding of the underlying biological mechanisms of PND. Alterations in inflammatory protein networks suggest distinct mechanisms between antenatal and postnatal depression. Application of biomarker tools, incorporating key proteins alongside patient history, could pave the way for personalised PND diagnosis and development of novel therapies. Presentation: Saturday, July 12, 2025

Disclosure: H.M. Hooker: None. D. Grammatopoulos: None. Pre-eclampsia (PE) is a multi-system pregnancy disorder and a leading cause of maternal and neonatal morbidity and mortality worldwide. Recent clinical practice for PE incorporates measurement of the ratio between two angiogenic markers, soluble fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF) in blood. Abnormal levels of sFlt-1 and PlGF are secreted from a dysfunctional placenta in PE into the maternal bloodstream, causing widespread maternal endothelial dysfunction. Whilst use of the ratio is a major advancement in early prediction and risk stratification for women with suspected PE, the biological mechanisms regulating the secretion patterns of these key angiogenic biomarkers remain poorly understood. This study aimed to implement a model of angiogenic biomarker secretion from placental trophoblasts, using the BeWo choriocarcinoma cell line, to examine the effects of potential regulators, focussing on corticotrophin releasing hormone (CRH), a major homeostatic regulator of fetomaternal communication, and glucose availability on the mRNA expression and secretion of sFlt-1 and PlGF. BeWo cells were differentiated into a syncytiotrophoblast-like state using 50μM forskolin for 48hrs, a cyclic AMP pathway activator. Forskolin treatment resulted in a substantial 3- to 4-fold increase in PlGF mRNA expression and Western blot analysis showed increased phosphorylation of cAMP-response element binding protein (CREB), a known PlGF transcription factor, indicating regulation of PlGF via the cAMP pathway. In contrast, a marginal increase in sFlt-1 mRNA was observed in forskolin-treated cells, suggesting no direct sFlt-1 control via cAMP in BeWo cells. Untreated BeWo cells exhibited a 250-fold greater expression of PlGF than sFlt-1, rising to 625-fold in forskolin-treated cells.CRH had little effect on undifferentiated cells but in differentiated cells CRH appears to promote PlGF expression and inhibit sFlt-1 expression. However, dose-response experiments identified that at higher concentrations, CRH effects may be diminished through downregulation of the CRH receptor. Moreover, in differentiated cells, an inverse relationship was observed between glucose availability and expression of PlGF and human chorionic gonadotrophin. In contrast, in both undifferentiated and differentiated cells, increased glucose availability resulted in a small but statistically significant increase in sFlt-1 expression. Overall, we have demonstrated significant PlGF expression in BeWo cells that appears to be under the regulation of placental hormones such as CRH as well as glucose availability. Understanding how these placental regulators affect the secretion of sFlt-1 and PlGF advances current knowledge of the pathogenesis of PE and how known risk factors such as gestational diabetes and high-stress pregnancies affect PE phenotype and sFlt-1/PlGF ratio levels. Presentation: Monday, July 14, 2025

Biomarkers of aging serve as important outcome measures in longevity-promoting interventions. However, there is limited consensus on which specific biomarkers are most appropriate for human intervention studies. This work aimed to address this need by establishing an expert consensus on biomarkers of aging for use in intervention studies via the Delphi method.A 3-round Delphi study was conducted using an online platform. In Round 1, expert panel members provided suggestions for candidate biomarkers of aging. In Rounds 2 and 3, they voted on 500 initial statements (yes/no) relating to 20 biomarkers of aging. Panel members could abstain from voting on biomarkers outside their expertise. Consensus was reached when there was >= 70% agreement on a statement/biomarker.Of the 460 international panel members invited to participate, 116 completed Round 1, 87 completed Round 2, and 60 completed Round 3. Across the 3 rounds, 14 biomarkers met consensus that spanned physiological (eg, insulin-like growth factor 1, growth-differentiating factor-15), inflammatory (eg, high sensitivity C-reactive protein, interleukin-6), functional (eg, muscle mass, muscle strength, hand grip strength, Timed-Up-and-Go, gait speed, standing balance test, frailty index, cognitive health, blood pressure), and epigenetic (eg, DNA methylation/epigenetic clocks) domains.Expert consensus identified 14 potential biomarkers of aging which may be used as outcome measures in intervention studies. Future aging research should identify which combination of these biomarkers has the greatest utility.

Disclosure: A. Aimaganova: None. N. Khovanova: None. E. Braybrook: None. D. Grammatopoulos: None. Thyroid dysfunction appears to be a common endocrinopathy in hospitalized COVID-19 patients. The most common pattern of deranged thyroid function tests (TFTs) reported is reminiscent of non-thyroidal illness (NTIS) or sick-euthyroid syndrome. NTIS is often observed in the critical care unit (CCU) setting, with a clear correlation to poor outcomes. The interdependence between the thyroid and immune system although recognized, is not fully understood. Decoding associations between biomarkers of inflammation and thyroid hormones in the context of disease severity may help to elucidate the adaptive mechanisms of the pituitary-thyroid (PT) axis and identify patterns of severity-specific thyroid dysfunction. In this single-center retrospective observational study, routine biomarker data incorporating TFTs were obtained from COVID patients, hospitalized between April 2020 and September 2022. Patients were classified as having severe COVID disease based on admission to CCU or general ward. Statistical tests based on the distribution of biomarkers were used to examine differences, Spearman rank correlations evaluated the strength of the associations, and correlation-based biomarker networks were developed. 237 data sets from 184 patients were extracted. 59% of data were from patients in CCU and 41% from wards. The mean age was 57 and 63 years respectively. Spearman correlations identified a negative relationship between markers of inflammation, red blood cell (RBC) parameters and iron metabolism, and a positive association between thyroid function, RBC parameters and iron metabolism. A deduced network, built around the correlation of TFTs, exhibited severity-specific characteristics. 11 biomarkers exhibited significant group-specific differences, including free triiodothyronine (fT3) and free thyroxine (fT4). An increased number of patients with fT3 below the reference range, in the context of normal TSH and fT4, were observed in those with severe disease. Correlations between components of the PT axis identified a unique interleukin-6 (IL-6) dependent enhancement of correlations between TSH and fT3 or fT3 to fT4 ratio, present mainly in CCU patients. Routine biomarkers have allowed the investigation of thyroid function dynamics and responses to disease signals, which can arise from distinct pathogenic patterns such as hyperinflammation. In place of singular correlation, we explored patterns of biomarker network correlations, which can provide unique information about an organism’s systemic responses. An increased number of correlations in CCU patients suggests activation of coordinated responses associated with enhanced disease severity. Our study has uncovered important differences in biomarker networks and thyroid hormone adaptive responses orchestrated by inflammatory signals such as IL-6, especially in critically ill patients. Presentation: Saturday, July 12, 2025

Objective: The mechanism of treatment-resistant hypertension (TRH) is not fully understood yet. However, studies suggest the involvement of vascular endothelial dysfunction and systemic inflammation as contributors to the pathogenesis. This study aims to identify any measurable differences in protein abundance between two groups of hypertensive adults, controlled hypertension (CH) and TRH, using proteomics analysis techniques. Design and method: The study samples were obtained from an existing observational cohort study involving CH and TRH groups; venous blood samples were previously stored at -80°C, and appropriate consenting and ethical approval was obtained. TRH was defined blood pressure (BP) of 140/90 mmHg or more, on 3 or more antihypertensive agents or controlled BP (less than140/90 mmHg) after taking 4 or more antihypertensive agents. The study was conducted in two phases: the discovery phase, where 60 samples of matched group's (CH, n=30, TRH n=30) Peripheral venous serum samples were initially depleted from the highly abundant proteins before undergoing trypsin digestion, this was followed by Liquid Chromatography Mass Spectrometry analysis. The validation phase included 140 candidates (CH n=81, TRH n=59). Results were statistically analysed using an independent T-test, P valve <0.05 was considered to be statistically significant. Gen Ontology (GO) description is used for the functional description of the proteins. Results: Full results analysis is still undergoing by the time this abstract is being submitted, however, 12 different proteins were found to be significantly different in their expression between the two groups: Alpha 1B glycoprotein, Leucine-rich alpha 2 glycoproteins, Inter alpha trypsin inhibitor heavy chain H3, Lumican, Complement component C9, Complement factor D, Contactin-1, Lysozyme C, Phospholipid transfer protein, and Vascular cell adhesion protein 1.Initial GO analysis suggests above proteins are involved in vascular integrity, endothelial function and inflammatory response. Conclusions: There are significant differences in the proteins expression between patients with TRH and CH. This may allow better understanding of TRH, and treatment strategies.

Background Pre-eclampsia remains a leading cause of maternal and foetal mortality with a poorly understood pathophysiology. It can lead to a range of clinical presentations, but proteinuria and hypertension are key components of the diagnosis. These signs arise due to disordered placental implantation due to poor trophoblastic invasion, resulting in placental oxidative stress due to hypoxia. Oxidative stress triggers the release of syncytiotrophoblast microvesicles (STMBs), of which placenta-derived exosomes may be a key component. The high specificity of exosomes for their cell of origin makes them ideal candidates as diagnostic biomarkers. We are particularly interested in the miRNAs (microRNAs) contained within these exosomes, as they may give us an insight into the genomic regulation within the pre-eclamptic placenta that leads to the disease state. The development of workflows for miRNA quantitation may enable us to identify novel biomarkers. Methods We extracted exosomes and purified total RNA from 23 serum samples using the Norgen Plasma/Serum Exosome Purification and RNA Isolation Midi Kit. We then used the bioanalyser to determine the concentration and quality of the RNA obtained. It uses rapid electrophoresis, requires minimal sample sizes, and can assess the quality of genetic material as small as 25 bases. Results We have successfully isolated RNA from these samples; however, the concentration of the total RNA was too low for downstream molecular analysis. We did gain insight into how to optimise and develop the workflow so that, with each attempt, the yield increased. Our greatest concentrations were obtained by combining serum samples from multiple patients, demonstrating that we needed a higher volume to optimise the yield. Future studies should aim to obtain samples specifically for use in this research so that we can process a larger volume of serum. Conclusions We have also noted that there is a positive correlation between the overall concentration of total RNA and a high sFlt-1/PlGF ratio. Preliminary analysis from Illumina identified with a high degree of confidence the presence of three miRNAs, namely, mir-498(46), mir-122(1), and mir-134(41). Further work is necessary to validate these findings and should focus on the possible future role of these miRNAs as biomarkers for the early diagnosis of pre-eclampsia.

IntroductionThe engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells, and, therefore, blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine elicited serum antibodies. The emergence of SARS-CoV-2 variants has necessitated the development of adaptable assays that can be applied to assess the effectiveness of antibody-based therapeutics.MethodsThrough the testing of a range of recombinant spike proteins, we have developed a cell-based, ACE2/spike protein interaction assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell-bound spike protein fluorescence.ResultsUsing spike proteins from the original "Wuhan" SARS-CoV-2 strain and the Delta and Omicron variants, we identified differential blocking activity of three monoclonal antibodies directed against the spike receptor-binding domain. Importantly, biological activity in the spike interaction assay translated to efficacy in a SARS-CoV-2 infection assay.DiscussionThe spike protein interaction assay can be used to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable for quantification of the impact of antibodies against new and emerging SARS-CoV-2 variants.

The hypothalamic type 2 corticotropin releasing hormone receptor (CRH-R2) plays critical roles in homeostatic regulation, particularly in fine tuning stress recovery. During acute stress, the CRH-R2 ligands CRH and urocortins promote adaptive responses and feeding inhibition. However, in rodent models of chronic stress, over-exposure of hypothalamic CRH-R2 to its cognate agonists is associated with urocortin 2 (Ucn2) resistance; attenuated cAMP-response element binding protein (CREB) phosphorylation and increased food intake. The molecular mechanisms involved in these altered CRH-R2 signalling responses are not well described. In the present study, we used the adult mouse hypothalamus-derived cell line mHypoA-2/30 to investigate CRH-R2 signalling characteristics focusing on gene expression of molecules involved in feeding and circadian regulation given the role of clock genes in metabolic control. We identified functional CRH-R2 receptors expressed in mHypoA-2/30 cells that differentially regulate CREB and AMP-activated protein kinase (AMPK) phosphorylation and downstream expression of the appetite-regulatory genes proopiomelanocortin (Pomc) and neuropeptide Y (Npy) in accordance with an anorexigenic effect. We studied for the first time the effects of Ucn2 on clock genes in native and in a circadian bioluminescence reporter expressing mHypoA-2/30 cells, detecting enhancing effects of Ucn2 on mRNA levels and rhythm amplitude of the circadian regulator Aryl hydrocarbon receptor nuclear translocator-like protein 1 (Bmal1), which could facilitate anorexic responses in the activity circadian phase. These data uncover novel aspects of CRH-R2 hypothalamic signalling that might be important in regulation of circadian feeding during stress responses.

Among the modulatory functions of thyrotropin-releasing hormone (TRH), an anorectic behavior in rodents is observed when centrally injected. Hypothalamic paraventricular nucleus (PVN) neurons receive serotonergic inputs from dorsal raphe nucleus and express serotonin (5HT) receptors such as 5HT1A, 5HT2A/2C, 5HT6, which are involved in 5HT-induced feeding regulation. Rats subjected to dehydration-induced anorexia (DIA) model show increased PVN TRH mRNA expression, associated with their decreased food intake. We analyzed whether 5HT input is implicated in the enhanced PVN TRH transcription that anorectic rats exhibit, given that 5HT increases TRH expression and release when studiedin vitro. By using mHypoA-2/30 hypothalamic cell cultures, we found that 5HT stimulated TRH mRNA, pCREB, and pERK1/2 levels. By inhibiting basal PKA or PKC activities or those induced by 5HT, pCREB or pERK1/2 content did not increase suggesting involvement of both kinases in their phosphorylation. 5HT effect on TRH mRNA was not affected by PKA inhibition, but it diminished in the presence of PKCi suggesting involvement of PKC in 5HT-induced TRH increased transcription. This likely involves 5HT2A/2Cand the activation of alternative transduction pathways than those studied here. In agreement with thein vitrodata, we found that injecting 5HT2A/2Cantagonists into the PVN of DIA rats reversed the increased TRH expression of anorectic animals, as well as their decreased food intake; also, the agonist reduced food intake of hungry restricted animals along with elevated PVN TRH mRNA levels. Our results support that the anorectic effects of serotonin are mediated by PVN TRH in this model.

Fetal exposure in adverse environmental factors during intrauterine life can lead to various biological adjustments, affecting not only in utero development of the conceptus, but also its later metabolic and endocrine wellbeing. During human gestation, maternal bone turnover increases, as reflected by molecules involved in bone metabolism, such as vitamin D, osteocalcin, sclerostin, sRANKL, and osteoprotegerin; however, recent studies support their emerging role in endocrine functions and glucose homeostasis regulation. Herein, we sought to systematically review current knowledge on the effects of aforementioned maternal bone biomarkers during pregnancy on fetal intrauterine growth and metabolism, neonatal anthropometric measures at birth, as well as on future endocrine and metabolic wellbeing of the offspring. A growing body of literature converges on the view that maternal bone turnover is likely implicated in fetal growth, and at least to some extent, in neonatal and childhood body composition and metabolic wellbeing. Maternal sclerostin and sRANKL are positively linked with fetal abdominal circumference and subcutaneous fat deposition, contributing to greater birthweights. Vitamin D deficiency correlates with lower birthweights, while research is still needed on intrauterine fetal metabolism, as well as on vitamin D dosing supplementation during pregnancy, to diminish the risks of low birthweight or SGA neonates in high-risk populations.

The COVID-19 pandemic spotlighted the need for transformation of pathology and laboratory medicine (PALM) to better meet public health-care needs. The Lancet Commission on Diagnostics1Fleming KA Horton S Wilson ML et al.The Lancet Commission on diagnostics: transforming access to diagnostics.Lancet. 2021; 398: 1997-2050Summary Full Text Full Text PDF PubMed Scopus (80) Google Scholar articulated recommendations around a fit-for-purpose global infrastructure; these are also relevant for the UK diagnostics strategy. Effective engagement with front-line PALM services and professional organisations is essential for the uptake of these recommendations and the introduction of transformative changes at the necessary pace and scale. For many years, UK diagnostics has focused on efficiencies gained from networking and service reconfiguration. Benefits from networking have emerged during the COVID-19 pandemic. We expect that contributions made during the pandemic will catalyse a shift in focus and mindset in our future approach to diagnostics; the case is indeed compelling. During the pandemic, PALM services have faced unpredictable clinical, analytical, and organisational challenges. Successful adaptation was based on learning to act in novel ways and set up new infrastructure to address urgent clinical care needs. This response involved mobilising academic and industry partners to capture and quickly translate innovation into clinical practice. Many services made multifaceted clinical and scientific contributions for patient benefit. For example, capacity testing gaps and urgent clinical care bottlenecks in managing admissions were addressed by dedicated high-throughput COVID-19 testing laboratories. The ability to successfully redeploy staff from different specialties competent in molecular methods highlights the need for highly skilled and flexible staff. PALM services with sufficient staff capacity contributed to national programmes by setting up specialist services, such as RNA sequencing to support the COVID-19 Genomics UK Consortium,2COVID-19 Genomics UKAn integrated national scale SARS-CoV-2 genomic surveillance network.Lancet Microbe. 2020; 1: e99-100Summary Full Text Full Text PDF PubMed Scopus (125) Google Scholar and supporting the UK National Health Service (NHS) Test and Trace technical validation groups. Rapid translation of novel biomarker concepts and timely implementation of recommended clinical investigations, such as those published by the Royal College of Pathologist3The Royal College of PathologistsGuidance on the use and interpretation of clinical biochemistry tests in patients with COVID-19 infection. The Royal College of Pathologists, London2020Google Scholar and BMJ Best Practice, became crucial for the effective management of patients with COVID-19, a particular challenge given the paucity of evidence-based markers to monitor progression and severity. We recognise the commitment of PALM services in the face of a global health-care emergency to contribute through the development of dedicated and accredited biobanks, artificial intelligence-based innovations, and medical technology solutions. However, most of these services have had chronic scientific-expertise depletion, making innovation and adoption a challenging goal. Nevertheless, mature partnerships with academia and industry have been a major driver to NHS-led innovation. This collaboration identifies a major focus on the transformation roadmap and makes a strong case for strengthening such collaborative interactions in the future. Planning for pandemic preparedness and tackling innovation in precision medicine and integrated care, requires the UK diagnostics landscape to optimise services cooperation and integration, and accelerate collaboration. Aligning with this vision will ensure that appropriate lessons from this pandemic have been learnt. LY is a member of the Medical and Scientific Advisory Board at Abingdon Health. NRA was president of the UK Association for Clinical Biochemistry and Laboratory Medicine. DKG declares no competing interests. The Lancet Commission on diagnostics: transforming access to diagnosticsAt the end of 2019, the first reports of a new respiratory virus appeared in China. The subsequent COVID-19 pandemic has affected every person, in every country, in the world. One early lesson was the crucial importance of timely accurate diagnosis. A second lesson was the widespread scarcity of such diagnostic capacity and capability. Full-Text PDF

The COVID-19 pandemic has highlighted the need for innovative biosensing, diagnostic, and surveillance platforms. Here we report that glycosylated, polymer-stabilized, gold nanorods can bind the SARS-CoV-2 spike protein and show correlation to the presence of SARS-CoV-2 in primary COVID-19 clinical samples. Telechelic polymers were prepared by reversible addition–fragmentation chain-transfer polymerization, enabling the capture of 2,3-sialyllactose and immobilization onto gold nanorods. Control experiments with a panel of lectins and a galactosamine-terminated polymer confirmed the selective binding. The glycosylated rods were shown to give dose-dependent responses against recombinant truncated SARS-CoV-2 spike protein, and the responses were further correlated using primary patient swab samples. The essentiality of the anisotropic particles for reducing the background interference is demonstrated. This highlights the utility of polymer tethering of glycans for plasmonic biosensors of infection.

The engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells and accordingly blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine-elicited serum antibodies. The emergence of SARS-CoV-2 variants necessitates the development of adaptable assays that can be applied to assess the effectiveness of therapeutics. Through testing of a range of recombinant spike proteins, we have developed a cell based, ACE2/spike protein binding assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell bound spike protein fluorescence. Using spike proteins from the original ‘Wuhan’ SARS-CoV-2 virus, as well as the delta and omicron variants, we identify differential blocking activity of three monoclonal antibodies directed against the spike receptor binding domain. Importantly, biological activity in the spike binding assay translated to efficacy in a SARS-CoV-2 infection assay. Hence, the spike binding assay has utility to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable to quantify impact of antibodies against new and emerging SARS-CoV-2 variants.

Early life is a period of considerable plasticity and vulnerability and insults during that period can disrupt the homeostatic equilibrium of the developing organism, resulting in adverse developmental programming and enhanced susceptibility to disease. Fetal exposure to prenatal stress can impede optimum brain development and deranged mother’s hypothalamic–pituitary–adrenal axis (HPA axis) stress responses can alter the neurodevelopmental trajectories of the offspring. Corticotropin-releasing hormone (CRH) and glucocorticoids, regulate fetal neurogenesis and while CRH exerts neuroprotective actions, increased levels of stress hormones have been associated with fetal brain structural alterations such as reduced cortical volume, impoverishment of neuronal density in the limbic brain areas and alterations in neuronal circuitry, synaptic plasticity, neurotransmission and G-protein coupled receptor (GPCR) signalling. Emerging evidence highlight the role of epigenetic changes in fetal brain programming, as stress-induced methylation of genes encoding molecules that are implicated in HPA axis and major neurodevelopmental processes. These serve as molecular memories and have been associated with long term modifications of the offspring’s stress regulatory system and increased susceptibility to psychosomatic disorders later in life. This review summarises our current understanding on the roles of CRH and other mediators of stress responses on fetal neurodevelopment.

The COVID-19 pandemic, and future pandemics, require diagnostic tools to track disease spread and guide the isolation of (a)symptomatic individuals. Lateral-flow diagnostics (LFDs) are rapid and of lower cost than molecular (genetic) tests, with current LFDs using antibodies as their recognition units. Herein, we develop a prototype flow-through device (related, but distinct to LFDs), utilizing N-acetyl neuraminic acid-functionalized, polymer-coated, gold nanoparticles as the detection/capture unit for SARS-COV-2, by targeting the sialic acid-binding site of the spike protein. The prototype device can give rapid results, with higher viral loads being faster than lower viral loads. The prototype's effectiveness is demonstrated using spike protein, lentiviral models, and a panel of heat-inactivated primary patient nasal swabs. The device was also shown to retain detection capability toward recombinant spike proteins from several variants (mutants) of concern. This study provides the proof of principle that glyco-lateral-flow devices could be developed to be used in the tracking monitoring of infectious agents, to complement, or as alternatives to antibody-based systems.

ABSTRACTControl of SARS-CoV-2 transmission is complicated by the emergence of variants, especially those containing mutations in the spike protein. By enhancing infectivity and evading immunity, infection with these variants might result in more severe clinical outcomes as well as being more resistant to vaccines developed on the basis of the original prototypic virus variant. One such example is the alpha variant (B.1.1.7), which has been detected in more than 100 countries and rapidly become the dominant strain in the UK in late 2020 and early 2021. There is an urgent need to develop appropriate surveillance programmes to rapidly monitor the spread of variants and to better understand the role of variants in disease outcomes and immune evasion. The nucleotide sequencing method, the ‘gold standard’ of variant detection, is unsuitable as a fast-response surveillance tool by frontline diagnostic services which require detection methods with short turnaround times. We developed a screening protocol based of sequential allele-specific qPCR for detection of the N501Y mutation and H69/V70 deletion present in the alpha/B.1.1.7 variant. We tested this protocol in previously confirmed positive samples from the Pathology Dept, University Hospital Coventry and Warwickshire during the second wave period in the UK (December 2020-March 2021). In these samples variant identity was confirmed by NGS sequencing via COG-UK. Our results identified increased incidence of variants containing both N501Y and Δ69/70 HV mutations, especially in patients admitted during January and early February 2021. This approach, which yields results within 3 hours, can be used as an initial rapid screening step with NGS as confirmatory follow-up. We also report that the increased prevalence of alpha/B.1.1.7 variant in admitted patients since mid-January 2021, a period that characterised peaked mortality rates, was associated with a sharp 2.5-fold rise in the mean circulating IL-6 level and to a lesser extent Troponin-T. More detailed biomarker analysis of a small cohort of patients (n=83), where variant status and clinical outcomes were available, demonstrated that deceased patients infected with the alpha/B.1.1.7 variant had significantly higher levels of inflammation and cell injury markers, especially IL-6 and LDH, compared to deceased patients infected with a non-alpha/B.1.1.7 variant, pointing towards a more severe inflammatory disease phenotype. In contrast, both groups survivors most biomarker exhibited levels below the group average, with distinct patterns of modified z-scores present.