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Background: Component‐resolved diagnosis allows detection of IgE sensitization having the advantage of reproducibility and standardization compared to crude extracts. The main disadvantage of the traditional allergen identification methods, 1‐ or 2‐dimensional western blotting and screening of expression cDNA libraries with patients' IgEs, is that the native structure of the protein is not necessarily maintained. Methods: We used a novel immunoprecipitation technique in combination with mass spectrometry to identify new allergens of Aspergillus fumigatus. Magnetic Dynabeads coupled with anti‐human IgE antibodies were used to purify human serum IgE and subsequently allergens from A. fumigatus protein extract. Results: Of the 184 proteins detected by subsequent mass peptide fingerprinting, a subset of 13 were recombinantly expressed and purified. In a panel of 52 A. fumigatus‐sensitized people with asthma, 23 non‐fungal‐sensitized asthmatics and 18 healthy individuals, only the former showed an IgE reaction by immunoblotting and/or ELISA. We discovered 11 proteins not yet described as A. fumigatus allergens, with fructose‐bisphosphate aldolase class II (FBA2) (33%), NAD‐dependent malate dehydrogenase (31%) and Cu/Zn superoxide dismutase (27%) being the most prevalent. With respect to these three allergens, native versus denatured protein assays indicated a better recognition of the native proteins. Seven of 11 allergens fulfilled the WHO/IUIS criteria and were accepted as new A. fumigatus allergens. Conclusion: In conclusion, we introduce a straightforward method of allergen identification from complex allergenic sources such as A. fumigatus by immunoprecipitation combined with mass spectrometry, which has the advantage over traditional methods of identifying allergens by maintaining the structure of the proteins. [ABSTRACT FROM AUTHOR]

The early detection of relapse following primary surgery for non-small-cell lung cancer and the characterization of emerging subclones, which seed metastatic sites, might offer new therapeutic approaches for limiting tumour recurrence. The ability to track the evolutionary dynamics of early-stage lung cancer non-invasively in circulating tumour DNA (ctDNA) has not yet been demonstrated. Here we use a tumour-specific phylogenetic approach to profile the ctDNA of the first 100 TRACERx (Tracking Non-Small-Cell Lung Cancer Evolution Through Therapy (Rx)) study participants, including one patient who was also recruited to the PEACE (Posthumous Evaluation of Advanced Cancer Environment) post-mortem study. We identify independent predictors of ctDNA release and analyse the tumour-volume detection limit. Through blinded profiling of postoperative plasma, we observe evidence of adjuvant chemotherapy resistance and identify patients who are very likely to experience recurrence of their lung cancer. Finally, we show that phylogenetic ctDNA profiling tracks the subclonal nature of lung cancer relapse and metastasis, providing a new approach for ctDNA-driven therapeutic studies., Author(s): Christopher Abbosh [1]; Nicolai J. Birkbak [1, 2]; Gareth A. Wilson [1, 2]; Mariam Jamal-Hanjani [1]; Tudor Constantin [3]; Raheleh Salari [3]; John Le Quesne [4]; David A. Moore [...]

The study aims to investigate whether Mepolizumab therapy is effective in reducing exacerbations among patients with refractory eosinophilic asthma. The results indicate that therapy with Mepolizumab is quite effective in reducing exacerbations amongst asthmatic patients .

Aim: To study sputum mediator profiles pattern in children with acute severe asthma, compared with stable asthma and healthy controls. The mechanisms of acute severe asthma attacks, such as biomarkers cascades and immunological responses, are poorly understood. Methods: We conducted a prospective observational case‐control study of children aged 5 to 17 years, who presented to hospital with an asthma attack. Children with stable asthma were recruited during outpatient asthma clinic visits. Control children without an asthma diagnosis were recruited from surgical wards. Sputum mediator profiles were measured, and sputum leukocyte differential cell counts were generated. Results: Sputum data were available in 48 children (acute asthma; n = 18, stable asthma; n = 17, healthy controls; n = 13). Acute‐phase biomarkers and neutrophil attractants such as IL‐6 and its receptor, IL‐8 and cytokines linked with bacterial signals, including TNF‐R1 and TNF‐R2, were elevated in asthma attacks versus stable asthma and healthy controls. T‐cell attractant cytokines, associated with viral infections, such as CCL‐5, CXCL‐10 and CXCL‐11, and CXCL‐9 (secreted from eosinophils after a viral trigger) were also raised. Conclusion: Mediator profiles consistent with bacterial and viral respiratory infections, and T2 inflammation markers co‐exist in the sputum of children with acute severe asthma attacks. [ABSTRACT FROM AUTHOR]

Abstract Background Severe asthma is a heterogeneous disease and the relationship between airway inflammation and airway remodelling is poorly understood. We sought to define sputum mediator profiles in severe asthmatics categorised by CT-determined airway geometry and sputum differential cell counts. Methods In a single centre cross-sectional observational study we recruited 59 subjects with severe asthma that underwent sputum induction and thoracic CT. Quantitative CT analysis of the apical segment of the right upper lobe (RB1) was performed. Forty-one mediators in sputum samples were measured of which 21 mediators that were assessable in >50% of samples were included in the analyses. Results Independent of airway geometry, sputum MMP9 and IL-1β were elevated in those groups with a high sputum neutrophil count while sputum ICAM was elevated in those subjects with a low sputum neutrophil count. In contrast, sputum CCL11, IL-1α and fibrinogen were different in groups stratified by both sputum neutrophil count and airway geometry. Sputum CCL11 concentration was elevated in subjects with a low sputum neutrophil count and high luminal and total RB1 area, whereas sputum IL1α was increased in subjects with a high sputum neutrophil count and low total RB1 area. Sputum fibrinogen was elevated in those subjects with RB1 luminal narrowing and in those subjects with neutrophilic inflammation without luminal narrowing. Conclusions We have demonstrated that sputum mediator profiling reveals a number of associations with airway geometry. Whether these findings reflect important biological phenotypes that might inform stratified medicine approaches requires further investigation.

Background: Sensitization to thermotolerant fungi, including filamentous fungi and Candida albicans, is associated with poor lung function in adults with severe asthma. Data in children are lacking. Environmental exposure to fungi is linked with acute severe asthma attacks, but there are few studies reporting the presence of fungi in the airways during asthma attacks. Methods: We investigated the association between fungal sensitization and/or positive fungal sputum culture and markers of asthma severity in children with chronic and acute asthma. Sensitization was determined using serum‐specific IgE and skin prick testing against a panel of five fungi. Fungal culture was focused towards detection of filamentous fungi from sputum samples. Results: We obtained sensitization data and/or sputum from 175 children: 99 with chronic asthma, 39 with acute asthma and 37 controls. 34.1% of children with chronic asthma were sensitized to thermotolerant fungi compared with no children without asthma (p =< 0.001). These children had worse pre‐bronchodilator lung function compared with asthmatics without sensitization including a lower FEV1/FVC ratio (p <.05). The isolation rate of filamentous fungi from sputum was higher in children with acute compared with chronic asthma. Conclusions: Fungal sensitization is a feature of children with chronic asthma. Children sensitized to thermotolerant fungi have worse lung function, require more courses of systemic corticosteroids and have greater limitation of activities due to asthma. Asthma attacks in children were associated with the presence of filamentous fungi positive sputum culture. Mechanistic studies are required to establish whether fungi contribute directly to the development of acute asthma. [ABSTRACT FROM AUTHOR]

Abstract Background In asthma interleukin (IL)-13 is increased in the airway compared with non-asthmatic eosinophilic bronchitis. Whether this differential expression is specific to the airway or is more generalised is uncertain. Methods We sought to examine IL-13 expression in peripheral blood T-cells and eosinophils in asthma and non-asthmatic eosinophilic bronchitis. Peripheral blood CD3+ cell and eosinophil intracellular IL-13 expression from subjects with asthma, non-asthmatic eosinophilic bronchitis and healthy controls was assessed. The effect of priming by asthmatic serum on the release of IL-13 by peripheral blood mononuclear cells from healthy subjects was examined and the serum from these subjects was analysed for a range of chemokines and cytokines. Results The median (IQR)% intracellular IL-13 expression by CD3+ cells was increased in asthma [5.3 (2.7–9.8)%; n = 12] compared to non-asthmatic eosinophilic bronchitis [1.1 (0.5–3)%; n = 7] and healthy controls [1.7 (0.2–3%); n = 9] (p = 0.02), but was not significantly different in eosinophils across the groups. IL-13 released from healthy peripheral blood mononuclear cells (n = 10) was increased by asthmatic serum [117 (47.8–198)pg/ml] compared to control [78.5 (42.6–128)pg/ml; p = 0.02), but was not affected by non-asthmatic serum. Conclusion Our findings support the view that IL-13 expression is increased in peripheral blood-derived T cells in asthma and that asthmatic serum up-regulates IL-13 release from healthy peripheral blood mononuclear cells.

Background: Fungal involvement in asthma is associated with severe disease. The full spectrum of fungal species in asthma is not well described and is derived largely from insensitive culture techniques. Objectives: To use high‐throughput sequencing to describe the airway mycobiota in asthmatics with and without fungal sensitization and healthy controls; to compare samples representing different airway compartments; to determine whether the mycobiota was influenced by the fungal composition of outdoor air; and to compare findings with clinically relevant outcomes. Methods: We amplified the internal transcribed spacer region 2 of the nuclear ribosomal operon to identify the fungal species present. Ninety‐seven subjects were recruited and provided sputum (83 asthmatics; 14 healthy subjects), with 29 also undergoing a bronchoscopy. A subset of airway samples were compared with matched outdoor air and mouthwash samples. Results: Two hundred and six taxa at the species level were identified in sputum, most at low relative abundance. Aspergillus fumigatus, Candida albicans and Mycosphaerella tassiana had the highest relative abundances and were the most prevalent species across all subjects. The airway mycobiota consisted of a complex community with high diversity between individuals. Notable shifts in the balance of fungi detected in the lung were associated with asthma status, asthma duration and biomarkers of inflammation. Aspergillus tubingensis, a member of the Aspergillus niger species complex, was most prevalent from bronchoscopic protected brush samples and significantly associated with a low sputum neutrophilia. Cryptococcus pseudolongus, from the Cryptococcus humicola species complex, was more abundant from bronchoscopy samples than sputum, and differentially more abundant in asthma than health. Conclusions and Clinical Relevance: The airway mycobiota was dominated by a relatively small number of species, but was distinct from the oropharyngeal mycobiota and air samples. Members of the A. niger and C. humicola species complexes may play unexpected roles in the pathogenesis of asthma. [ABSTRACT FROM AUTHOR]

Background: Eosinophilia is frequently a feature of asthma. Sputum analysis can help with the diagnosis and phenotyping of asthma. The current gold standard method is unsuitable for samples <100 mg. However, children frequently produce samples below this threshold. Aim: To compare and validate our modified, small sample (>10 mg and <100 mg) sputum processing method (which omits sample filtering), with the current gold standard. Method: Prospective study of 32 adults with severe asthma providing sputum samples of sufficient size for dual processing. Results: The median (IQR) sample weight was 211.0 (162.4–185.5) mg and 57.5 (22.0–61.6) mg for standard, and small sputum sample processing respectively. There was no statistically significant difference in the median (IQR) cell counts between Method A and B, respectively: eosinophils 3.8% (1.5–14.0) versus 4.9% (1.3–15.5); neutrophils 78.1% (46.5–92.4) versus 65.0% (48.3–86.6). Conclusion: The small sputum sample processing is feasible and reliable, and yields similar results to standard processing. [ABSTRACT FROM AUTHOR]

Summary Background Patients with chronic obstructive pulmonary disease (COPD) often have vitamin D deficiency, which is associated with increased susceptibility to upper respiratory infection—a major precipitant of exacerbation. Multicentre trials of vitamin D supplementation for prevention of exacerbation and upper respiratory infection in patients with COPD are lacking. We therefore investigated whether vitamin D 3 (colecalciferol) supplementation would reduce the incidence of moderate or severe COPD exacerbations and upper respiratory infections. Methods We did a randomised, double-blind, placebo-controlled trial of vitamin D 3 supplementation in adults with COPD in 60 general practices and four Acute National Health Service Trust clinics in London, UK. Patients were allocated to receive six 2-monthly oral doses of 3 mg vitamin D 3 or placebo over 1 year in a 1:1 ratio using computer-generated permuted block randomisation. Participants and study staff were masked to treatment assignment. Coprimary outcomes were time to first moderate or severe exacerbation and first upper respiratory infection. Analysis was by intention to treat. A prespecified subgroup analysis was done to assess whether effects of the intervention on the coprimary outcomes were modified by baseline vitamin D status. This trial is registered with ClinicalTrials.gov , number NCT00977873 . Findings 240 patients were randomly allocated to the vitamin D 3 group (n=122) and placebo group (n=118). Vitamin D 3 compared with placebo did not affect time to first moderate or severe exacerbation (adjusted hazard ratio 0·86, 95% CI 0·60–1·24, p=0·42) or time to first upper respiratory infection (0·95, 0·69–1·31, p=0·75). Prespecified subgroup analysis showed that vitamin D 3 was protective against moderate or severe exacerbation in participants with baseline serum 25-hydroxyvitamin D concentrations of less than 50 nmol/L (0·57, 0·35–0·92, p=0·021), but not in those with baseline 25-hydroxyvitamin D levels of at least 50 nmol/L (1·45, 0·81–2·62, p=0·21; p=0·021 for interaction between allocation and baseline serum 25-hydroxyvitamin D status). Baseline vitamin D status did not modify the effect of the intervention on risk of upper respiratory infection (p interaction =0·41). Interpretation Vitamin D 3 supplementation protected against moderate or severe exacerbation, but not upper respiratory infection, in patients with COPD with baseline 25-hydroxyvitamin D levels of less than 50 nmol/L. Our findings suggest that correction of vitamin D deficiency in patients with COPD reduces the risk of moderate or severe exacerbation. Funding UK National Institute for Health Research. [ABSTRACT FROM AUTHOR]

Background. Asthma and obesity are common; however the impact of obesity upon asthma remains uncertain. Objectives. To assess relationships between obesity and fat mass with airway inflammation, lung function, and disease control in patients with refractory asthma. Methods. 151 refractory asthma patients were characterised for measures of airway inflammation, lung function, Juniper asthma control questionnaire (JACQ), body mass index (BMI), and fat mass index (FMI) derived from dual energy X-ray absorptiometry. Patients were reassessed over 12 months. Results. 74% of patients had an elevated BMI. BMI and FMI correlated (r = 0.9, P < .001). FMI and JACQ correlated in men (r = 0.3, P = .01). After 12 months 23% lost weight. Weight change over 12 months correlated with FEV1 change (r = −0.3, P = .03), but not with change in JACQ or exacerbations. Conclusion. Increased fat mass is common in refractory asthma and is associated with asthma symptom control in men. Loss of weight is associated with improvement in lung function in refractory asthma. [ABSTRACT FROM AUTHOR]

Among 307 adults with OCD, early onset (age 5-15 years) was more common in men and later onset (age 26-35 years) in women. Early onset was associated with more checking, and late onset with more washing. More women than men had a history of treated depression; 12% of the women but none of the men had a history of anorexia. More women than men were married. Gender-divergent features may reflect differential aetiological factors. Our sample resembled others in the literature in its slight overall female preponderance, low rate of marriage and low fertility, onset mainly before age 35 years, chronicity, and common present and past depression. [ABSTRACT FROM AUTHOR]

Background: Cancer treatments may compromise fertility and family building in reproductive-age women. Previous research has shown that younger women with cancer experienced several reproductive health concerns., Objective: The aim of this study was to conduct a focused review of existing research about the subjective perceptions of reproductive concerns among young women with cancer (aged 15-49 years) and identify their potential predictors and outcomes., Methods: A systematic synthesis of mixed-methods research was conducted including peer-reviewed articles in English. Relevant studies were identified through the electronic databases Scopus, Web of Science, PubMed, ProQuest, and Psychology & Behavioral Sciences Collection (through EBSCOhost)., Results: A total of 22 reports met the eligibility criteria (8 qualitative and 14 quantitative). Research showed that younger women reported concerns related to their fertility status and/or own health after conception, their children's health, and their dyadic relationships. Redefinition of the motherhood role and the family future were also a source of concern. However, there is variance among women in concerns and experiences based on life stages and expectations., Conclusions: Reproductive concerns seem to be affected by personal circumstances and previous therapeutics. These concerns constitute a potential risk factor, simultaneously, for psychosocial maladjustment and adherence to endocrine therapy and fertility care., Implications for Practice: This article proposes a conceptual framework to understand the dimensions and potential predictors and outcomes of perceived concerns among reproductive-age cancer patients. Our data allow us to look at these concerns from a multifactorial perspective, identifying areas to be addressed in providing clinical care, namely, by nurses accompanying patients over an extended period., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2020 Wolters Kluwer Health, Inc. All rights reserved.)

Background: In asthma interleukin (IL)-13 is increased in the airway compared with non-asthmatic eosinophilic bronchitis. Whether this differential expression is specific to the airway or is more generalised is uncertain., Methods: We sought to examine IL-13 expression in peripheral blood T-cells and eosinophils in asthma and non-asthmatic eosinophilic bronchitis. Peripheral blood CD3+ cell and eosinophil intracellular IL-13 expression from subjects with asthma, non-asthmatic eosinophilic bronchitis and healthy controls was assessed. The effect of priming by asthmatic serum on the release of IL-13 by peripheral blood mononuclear cells from healthy subjects was examined and the serum from these subjects was analysed for a range of chemokines and cytokines., Results: The median (IQR)% intracellular IL-13 expression by CD3+ cells was increased in asthma [5.3 (2.7-9.8)%; n = 12] compared to non-asthmatic eosinophilic bronchitis [1.1 (0.5-3)%; n = 7] and healthy controls [1.7 (0.2-3%); n = 9] (p = 0.02), but was not significantly different in eosinophils across the groups. IL-13 released from healthy peripheral blood mononuclear cells (n = 10) was increased by asthmatic serum [117 (47.8-198)pg/ml] compared to control [78.5 (42.6-128)pg/ml; p = 0.02), but was not affected by non-asthmatic serum., Conclusion: Our findings support the view that IL-13 expression is increased in peripheral blood-derived T cells in asthma and that asthmatic serum up-regulates IL-13 release from healthy peripheral blood mononuclear cells.