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Biological sex is a crucial, but poorly understood variable in age‐related susceptibility to infection. Monocytes are important immune cells responsible for initiating and resolving inflammatory responses to infection. While changes in monocyte populations result in increased susceptibility to infection, there is limited research on the impact of age and sex on human monocyte phenotype and function. The aim of this work was to dissect the impact of increasing age and biological sex on human monocyte phenotype and function. Here, we show that older females have increased inflammatory intermediate and non‐classical monocytes compared to young. These monocyte subsets were the most inflammatory ex vivo, and their frequency correlated with markers of inflammageing. Proteomic analysis of sorted monocyte populations demonstrated that the three human monocyte subsets have largely distinct phenotypes. Key age‐associated protein pathways were identified, including complement cascade and phagocytosis. We confirmed the proteomics findings, showing that circulating C3 concentrations were reduced with age in females but not males. This decrease in complement in older females resulted in reduced monocyte phagocytosis. Crucially, we demonstrate that in peri/menopausal females, hormone replacement therapy (HRT) reversed this expansion in intermediate monocytes and decreased circulating CRP as compared to age‐matched controls. Importantly, peri/menopausal females on HRT had increased C3 serum concentrations and significant improvement in monocyte phagocytosis. The data presented here indicate the importance of menopause in aging monocyte phenotype and function. These data highlight the potential use of HRT in restoring monocyte function in females during aging and potentially improving anti‐pathogen immunity. Post‐menopausal females (> 64 years) have an increase in frequency of inflammatory monocyte populations with defective phagocytosis as compared to pre‐menopausal (< 40 years) females. Hormone Replacement Therapy (HRT) use in menopausal females (44–60 years) decreases the frequency of inflammatory monocytes and improves phagocytosis as compared to age‐matched controls.

$\\bullet$ Nitrogen (N) is known to be transferred from fungus to plant in the arbuscular mycorrhizal (AM) symbiosis, yet its metabolism, storage and transport are poorly understood. $\\bullet$ In vitro mycorrhizas of Glomus intraradices and Ri T-DNA-transformed carrot roots were grown in two-compartment Petri dishes. $^{15}N- and/or ^{13}C-labeled$ substrates were supplied to either the fungal compartment or to separate dishes containing uncolonized roots. The levels and labeling of free amino acids (AAs) in the extraradical mycelium (ERM) in mycorrhizal roots and in uncolonized roots were measured by gas chromatography/mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). $\\bullet$ Arginine (Arg) was the predominant free AA in the ERM, and almost all Arg molecules became labeled within 3 wk of supplying 15NH4+ to the fungal compartment. Labeling in Arg represented > 90% of the total 15N in the free AAs of the ERM. $\\lbrack Guanido-2-^{15}N\\rbrack Arg$ taken up by the ERM and transported to the intraradical mycelium (IRM) gave rise to $^{15}N-labeled$ AAs. $\\lbrack U-^{13}C\\rbrack Arg$ added to the fungal compartment did not produce any 13C labeling of other AAs in the mycorrhizal root. $\\bullet$ Arg is the major form of N synthesized and stored in the ERM and transported to the IRM. However, NH4+ is the most likely form of N transferred to host cells following its generation from Arg breakdown.

Both the plant and the fungus benefit nutritionally in the arbuscular mycorrhizal symbiosis: The host plant enjoys enhanced mineral uptake and the fungus receives fixed carbon. In this exchange the uptake, metabolism, and translocation of carbon by the fungal partner are poorly understood. We therefore analyzed the fate of isotopically labeled substrates in an arbuscular mycorrhiza (in vitro cultures of Ri T-DNA-transformed carrot [Daucus carota] roots colonized by Glomus intraradices) using nuclear magnetic resonance spectroscopy. Labeling patterns observed in lipids and carbohydrates after substrates were supplied to the mycorrhizal roots or the extraradical mycelium indicated that: (a) 13C-labeled glucose and fructose (but not mannitol or succinate) are effectively taken up by the fungus within the root and are metabolized to yield labeled carbohydrates and lipids; (b) the extraradical mycelium does not use exogenous sugars for catabolism, storage, or transfer to the host; (c) the fungus converts sugars taken up in the root compartment into lipids that are then translocated to the extraradical mycelium (there being little or no lipid synthesis in the external mycelium); and (d) hexose in fungal tissue undergoes substantially higher fluxes through an oxidative pentose phosphate pathway than does hexose in the host plant.

Indirect root for nitrogen The discovery of a previously unknown mechanism of nitrogen transfer from the arbuscular mycorrhizal fungi found on the roots of most land plants, to the host plants suggests that this symbiotic relationship may be a much more important factor in the global nitrogen cycle than was thought. The mechanism involves uptake of inorganic nitrogen by the fungus outside the roots, conversion to amino acids within the fungus, then transfer as ammonium ions from the fungal mycelium into the plant. The first event in host recognition by arbuscular mycorrhizal fungi is thought to be hyphal branching. A strigolactone, 5-deoxy-strigol, isolated from Lotus japonicus has now been identified as an inducer of branching. Strigolactones are root metabolites, previously isolated as seed germination stimulants for root parasitic weeds. This finding highlights the close relationship between plant and fungus, and may provide a new strategy for the control of both beneficial fungal symbionts and destructive parasitic weeds in agriculture and natural ecosystems. Most land plants are symbiotic with arbuscular mycorrhizal fungi (AMF), which take up mineral nutrients from the soil and exchange them with plants for photosynthetically fixed carbon. This exchange is a significant factor in global nutrient cycles 1 as well as in the ecology 2 , evolution 3 and physiology 4 of plants. Despite its importance as a nutrient, very little is known about how AMF take up nitrogen and transfer it to their host plants 5 . Here we report the results of stable isotope labelling experiments showing that inorganic nitrogen taken up by the fungus outside the roots is incorporated into amino acids, translocated from the extraradical to the intraradical mycelium as arginine, but transferred to the plant without carbon. Consistent with this mechanism, the genes of primary nitrogen assimilation are preferentially expressed in the extraradical tissues, whereas genes associated with arginine breakdown are more highly expressed in the intraradical mycelium. Strong changes in the expression of these genes in response to nitrogen availability and form also support the operation of this novel metabolic pathway in the arbuscular mycorrhizal symbiosis.

BackgroundMany patients with SARS-CoV-2 infection are reporting long lasting symptoms, requiring holistic multi-disciplinary rehabilitation; however, there are severe capacity restraints in rehabilitation services. We report on the development and initial experience of a digitally-enabled remote, supported rehabilitation programme: ‘Covid Recovery’ from UCLP/Living With.Methods Covid Recovery includes: (i) a clinical pathway; (ii) an app delivering physiotherapy, dietetic and psychology education and treatments for common symptoms (breathlessness, fatigue, anxiety) plus validated Patient-Reported Outcome Measures (PROMS); and (iii) a digital dashboard for clinicians to monitor patient activity and progress. A two-way messaging function allows personalisation of advice.ResultsIn the first 3 months, 66 patients with diverse demographics have been registered on the App: mean (range) age 54 years (23–78 years); 33 female; 19 Black/Asian ethnicity, 38 White/Caucasian ethnicity, remainder not stated/other. Amongst patients registered on the App for at least one week, patients undertake a mean average 7.0 actions per week covering ‘recording weight’, ‘completing a PROM – FACIT-F, Covid Recovery, GAD-7, MRC Breathlessness, D-12’, ‘tracking exercise’, and finishing a ‘’fatigue diary’. Overall on average each patient has read 11 articles, and 1 in 2 patients are creating and tracking a goal. Weekly review takes 2 – 3 minutes per patient.Example Case: A 47-year-old male joined the App following primary care managed Covid-19 but with residual symptoms at 12 weeks. In 77 days of using the App, he logged 97 activities including 15 messages on the two-way platform. Clinician support focused on managing breathlessness and fatigue symptoms whilst returning to exercise. Significant improvement was identified across the outcome measures for both physical and mental health in conjunction with an increase in exercise activity and intensity (see figure 1).Abstract L10 Figure 1Example patient FACIT-Fatigue outcomesDiscussionPatients recovering from Covid-19 report multiple and variable symptoms. Covid Recovery provides specialist services in a personalised manner, supporting patients through their rehabilitation and recovery journey, enabling them to feel seen, heard and believed.

Vesicular-arbuscular mycorrhizal fungi are symbionts for a large variety of crop plants; however, the form in which they take up carbon from the host is not established. To trace the course of carbon metabolism, we have used nuclear magnetic resonance spectroscopy with [13C]glucose labeling in vivo and in extracts to examine leek (Allium porrum) roots colonized by Glomus etunicatum (and uncolonized controls) as well as germinating spores. These studies implicate glucose as a likely substrate for vesicular-arbuscular mycorrhizal fungi in the symbiotic state. Root feeding of 0.6 mM 1-[13C]glucose labeled only the fungal metabolites trehalose and glycogen. The time course of this labeling was dependent on the status of the host. Incubation with 50 mM 1-[13C]glucose caused labeling of sucrose (in addition to fungal metabolites) with twice as much labeling in uncolonized plants. There was no detectable scrambling of the label from C1 glucose to the C6 position of glucose moieties in trehalose or glycogen. Labeling of mannitol C1,6 in the colonized root tissue was much less than in axenically germinating spores. Thus, carbohydrate metabolism of host and fungus are significantly altered in the symbiotic state

Environmental factors have a strong causal role in the development of asthma. Vitamin D insufficiency and particulate matter (PM) air pollution are two environmental factors associated with airways disease. We hypothesise that vitamin D will reduce production of inflammatory mediators by bronchial epithelial cells stimulated with PM, protecting the airways from inflammation that would otherwise promote Th2/Th17 responses in asthma. Primary human bronchial epithelial cells from healthy and asthmatic donors were cultured with standardised PM with or without vitamin D. Activated 1,25(OH)D3 was used in preliminary experiments and the precursor 25(OH)D3 in later experiments. A transcription microarray was conducted to highlight differentially expressed inflammatory mediators for further investigation. Expression of target genes was measured by quantitative real-time PCR and levels of inflammatory cytokines in culture supernatants by Cytometric Bead Array. PM caused increased production of multiple cytokines and chemokines by bronchial epithelial cells. Vitamin D decreased production of a range of cytokines and chemokines, including interleukin (IL) 6 (47·8% [95% CI 5·3%–90·3%] reduction in gene expression with 1,25(OH)D3, 46·7% [35·6%–57·8%] decrease in supernatant protein) and IL24 (61.3% [95% CI 36·0%–86.5%] reduction in gene expression). This reduction was not due to loss of cell viability. Other cytokines were not affected by vitamin D—for example, granulocyte-macrophage colony-stimulating factor (12·9% [95% CI −19·5% to 45·4%] reduction in gene expression, 1·9% [–16·7% to 20·5%] in supernatant protein). The decrease in IL-6 production on treating PM-stimulated bronchial cells with vitamin D, either active 1,25(OH)D3 or the circulating precursor 25(OH)D3, is important given that IL6 inhibits regulatory T-cell responses and promotes Th17 responses. IL24 promotes Th1 cytokine secretion. Functional studies investigating the effect of epithelial-conditioned medium on T cells are in progress. Wellcome Trust and National Institute for Health Research.

IntroductionBenralizumab is an anti-interleukin-5 receptor α monoclonal antibody indicated as an add-on maintenance therapy in adult patients with severe eosinophilic asthma (SEA). Herein we present real-data from the Benralizumab Patient Access Programme (BPAP) on the asthma-related, hospital healthcare resource utilisation (HCRU) in patients with severe asthma.MethodsThe BPAP study is a multi-centre, retrospective chart review study of patients with SEA from eight severe asthma centres in the UK. Data were collected from the medical records of patients receiving their first benralizumab dose between April 2018 and November 2019. Outcomes were assessed using descriptive statistics. HCRU including hospitalisations and emergency department (ED) visits related to exacerbations were described in the 12 months prior to benralizumab initiation (baseline), 1- and 2-years post benralizumab initiation. Patients remaining on treatment were included at each timepoint.ResultsA total of 276 patients were included. During baseline, 42% of patients had ≥1 asthma-related ED attendance; this proportion decreased to 23% at Year 2. Mean (SD) ED attendances reduced from 1.2 (2.2) at baseline to 0.4 (1.0) at Year 2, a relative reduction of 67%. The proportion of patients with ≥1 hospitalisation was 39% at baseline, falling to 18% in Year 2. Mean (SD) hospitalisations reduced from 1.0 (1.8) at baseline to 0.4 (1.0) at Year 2, a relative reduction of 70%. For patients with inpatient hospitalisations during the baseline with a recorded length of stay (n=50), median (IQR) length of stay was 6.5 (3.0–17.3) days; this decreased to 2.5 (0.0–8.3) days (n=38) at Year 2; a relative reduction of 62%. There was also a reduction in intensive care admissions from 16/241 (7%) at baseline to 5/209 (2%) at year 2.Abstract M25 Table 1Asthma-related healthcare resource utilization: ED attendances, hospitalisations and length of hospital stay (days) and ICU admissions at baseline, 1 and 2 years post benralizumab treatment HCRU Baseline 1 year 2 years ED visits Proportion of patients with a given number of visits No visits 137/237 (58%) 196/245 (80%) 160/209 (77%) 1 visit 36/237 (15%) 28/245 (11%) 32/209 (15%) 2 visits 26/237 (11%) 7/245 (3%) 7/209 (3%) ≥3 visits 38/237 (16%) 14/245 (6%) 10/209 (5%) Mean (SD) 1.2 (2.2) 0.2 (0.6) 0.4 (1.0) Median (IQR) 0.0 (0.0–2.0) 0.0 (0.0–0.0) 0.0 (0.0–0.0) Hospitalisations Proportion of patients with a given number of visits No visits 153/249 (61%) 208/245 (85%) 171/209 (82%) 1 visit 38/249 (15%) 23/245 (9%) 28/209 (13%) 2 visits 25/249 (10%) 5/245 (2%) 3/209 (1%) ≥3 visits 33/249 (13%) 9/245 (4%) 7/209 (3%) Mean (SD) 1.0 (1.8) 0.3 (0.9) 0.3 (0.9) Median (IQR) 0.0 (0.0–1.0) 0.0 (0.0–0.0) 0.0 (0.0–0.0) Length of hospitalisation stay (days)* Proportion of patients with a given number of days 0 days 2/50 (4%) 9/38 (24%) 10/39 (26%) 1 day 5/50 (10%) 4/38 (11%) 3/39 (8%) 2 days 3/50 (6%) 4/38 (11%) 6/39 (15%) ≥3 days 40/50 (80%) 21/38 (55%) 20/39 (51%) Mean (SD) 12.6 (13.4) 6.6 (10) 5.5 (8.2) Median (IQR) 6.5 (3.0–17.3) 3.0 (0.5–8.0) 2.5 (0.0–8.3) ICU admissions 16/241 (7%) 8/245 (3%) 5/209 (2%) *Denominator (n=50) is patients with length of hospitalisation recorded. Proportions may not sum to 100% due to rounding.ConclusionsThe results show that patients with SEA treated with benralizumab experienced clinically meaningful and sustained reductions in un-scheduled hospital HCRU. Treatment with benralizumab may be associated with >60% reduction in ED visits, hospitalisation and length of stay across all sites. This would reduce pressures on acute services and further work is warranted to investigate the overall economic impact of reduced HCRU.Please refer to page A295 for declarations of interest related to this abstract.

Introduction and ObjectivesBenralizumab is an anti-interleukin-5 receptor α monoclonal antibody indicated as an add-on maintenance treatment in adult patients with severe eosinophilic asthma (SEA). Herein we present real-world data from the Benralizumab Patient Access Programme (BPAP) for a subgroup of patients with SEA and concomitant nasal polyposis (SEAwNP) after 2 years of treatment with benralizumab for SEA.MethodsThe BPAP study is a multi-centre, retrospective, observational study of patients with SEA from eight UK centres. Data were collected from the medical records of patients receiving their first benralizumab dose between April 2018 and November 2019. Outcomes were assessed using descriptive statistics, with patients censored when treatment was discontinued.ResultsWithin the BPAP cohort of 276 patients, 57 (21%) patients had SEAwNP and were included in this subgroup analysis. The mean age was 50.9 (standard deviation [SD] 13.5); 46% (26/57) were female; mean BMI was 30.2 (SD 5.7); 61% (35/57) of patients were receiving maintenance oral corticosteroids (mOCS) at baseline. Clinical outcomes for the overall BPAP cohort and SEAwNP subgroup are shown in the table 1. Patients with SEAwNP had a mean annualised exacerbation rate (AER) of 3.8 (95% confidence interval [CI] 3.1-4.5) at baseline (n=56), decreasing to 0.8 (95% CI 0.5-1.1) at Year 2 (n=48); a relative reduction of 79%, with 44% of patients being exacerbation free over 2 years. For patients with SEAwNP on mOCS at baseline, 57% were off mOCS for asthma after 2 years, with 77% achieving a dose reduction of ≥50%. At baseline, mean asthma control questionnaire (ACQ-6) score was 2.9 (SD 1.5, n=55); this reduced to 1.3 (SD 1.5. n=30) at 2 years. The proportion of patients with SEAwNP achieving an improvement of ≥0.5 was 71% (20/28); 63% (19/30) had an ACQ-6 score of <1.5. Clinical outcomes in the overall BPAP cohort and SEAwNP subgroup were comparable (table 1).ConclusionsThis analysis in a subgroup of patients with SEAwNP shows that patients who were treated with benralizumab had clinically relevant and sustained improvements in clinical outcomes comparable to the overall cohort, including improved exacerbation rates, mOCS use and asthma symptom control.Abstract P130 Table 1Annualised exacerbation rate (AER), maintenance oral corticosteroid (mOCS) use and patient-reported outcome data (asthma control [ACQ-6]) in the BPAP sample, and in the subgroup of patients with SEA and concomitant NP Overall BPAP cohort Subgroup of patients with SEA and concomitant NP AER Baseline 2 years Baseline 2 years Mean (95%CI) AER a 5.3 (4.8–5.7) 1.1 (0.9–1.2) 3.8 (3.1– 4.5) 0.8 (0.5–1.1) Relative change in mean from baseline (%) - -79% - -79% Proportion exacerbation free (%) 27/273 (10%) 72/209 (34%) 8/56 (14%) 21/48 (44%) mOCS use Baseline 2 years Baseline 2 years Proportion on mOCS in overall cohort, n (%) a,b 174/276 (63%) 63/208 (30%) 35/57 (61%) 13/48 (27%) Proportion on mOCS in patients on mOCS at baseline, n (%) b - 57/127 (45%) - 13/30 (43%) mOCS dose (mg/day) in patients on mOCS at baseline, median (IQR) b 10.0 (5.0–20.0) 0.0 (0.0–5.0) 10.0 (5.0–15.0) 0.0 (0.0–5.0) Proportion of patients with ≥50% reduction in mOCS dose; n (%) [95% CI] - 93/127 (73%) [65%–81%] 23/30 (77%) [58%–90%] ACQ-6 Baseline 2 years Baseline 2 years Mean (SD) ACQ-6 score a 3.0 (1.5) 1.6 (1.5) 2.9 (1.5) 1.3 (1.5) Proportion of patients with improvement of ≥0.5 units for AQC-6 from baseline, n (%) a - 89/128 (70%) - 20/28 (71%) Proportion of patients ACQ-6 score of <1.5 44/257 (17%) 72/134 (54%) 12/55 (22%) 19/30 (63%) aCalculated for overall cohort (all patients with available data at that time point who remained on treatment) bCalculated for patients on mOCS (≥5 mg) at baseline onlyPlease refer to page A291 for declarations of interest related to this abstract.