Genomic epidemiology and longitudinal sampling of ward wastewater environments and patients reveals complexity of the transmission dynamics of -carbapenemase-producing Enterobacterales in a hospital setting.

Background: Healthcare-associated wastewater and asymptomatic patient reservoirs colonized by carbapenemase-producing Enterobacterales (CPE) contribute to nosocomial CPE dissemination, but the characteristics and dynamics of this remain unclear.

Methods: We systematically sampled wastewater sites ( = 4488 samples; 349 sites) and patients ( = 1247) across six wards over 6-12 months to understand bla-associated CPE (KPC-E) diversity within these reservoirs and transmission in a healthcare setting. Up to five KPC-E-positive isolates per sample were sequenced (Illumina).

Th2 high and mast cell gene signatures are associated with corticosteroid sensitivity in COPD.

Rationale: Severe asthma and chronic obstructive pulmonary disease (COPD) share common pathophysiological traits such as relative corticosteroid insensitivity. We recently published three transcriptome-associated clusters (TACs) using hierarchical analysis of the sputum transcriptome in asthmatics from the Unbiased Biomarkers for the Prediction of Respiratory Disease Outcomes (U-BIOPRED) cohort comprising one Th2-high inflammatory signature (TAC1) and two Th2-low signatures (TAC2 and TAC3).

Objective: We examined whether gene expression signatures obtained in asthma can be used to identify the subgroup of patients with COPD with steroid sensitivity.

Using Point of Care Testing to estimate influenza vaccine effectiveness in the English primary care sentinel surveillance network.

Introduction: Rapid Point of Care Testing (POCT) for influenza could be used to provide information on influenza vaccine effectiveness (IVE) as well as influencing clinical decision-making in primary care.

Methods: We undertook a test negative case control study to estimate the overall and age-specific (6 months-17 years, 18-64 years, ≥65 years old) IVE against medically attended POCT-confirmed influenza. The study took place over the winter of 2019-2020 and was nested within twelve general practices that are part of the Oxford-Royal College of General Practitioners (RCGP) Research and Surveillance Centre (RSC), the English sentinel surveillance network.

Peripheral airways type 2 inflammation, neutrophilia and microbial dysbiosis in severe asthma.

Background: IL-13 is considered an archetypal T2 cytokine central to the clinical disease expression of asthma. The IL-13 response genes, which are upregulated in central airway bronchial epithelial of asthma patients, can be normalized by high-dose inhaled steroid therapy in severe asthma. However, this is not the case within the peripheral airways.

Erratum: Blume, C., et al. Modulation of Human Airway Barrier Functions during and Infection Running Title: Airway Barrier Functions during Bacterial Infections. 2016, , 53.

There is an error in the title [...

Postnatal adaptations of phosphatidylcholine metabolism in extremely preterm infants: implications for choline and PUFA metabolism.

Background: Lipid metabolism in pregnancy delivers PUFAs from maternal liver to the developing fetus. The transition at birth to diets less enriched in PUFA is especially challenging for immature, extremely preterm infants who are typically supported by total parenteral nutrition.

Objective: The aim was to characterize phosphatidylcholine (PC) and choline metabolism in preterm infants and demonstrate the molecular specificity of PC synthesis by the immature preterm liver in vivo.

Integrating molecular point-of-care testing for influenza into primary care: a mixed-methods feasibility study.

Background: Molecular point-of-care testing (POCT) for influenza in primary care could influence clinical care and patient outcomes.

Aim: To assess the feasibility of incorporating influenza POCT into general practice in England.

Design And Setting: A mixed-methods study conducted in six general practices that had not previously participated in respiratory virology sampling, which are part of the Royal College of General Practitioners Research and Surveillance Centre English sentinel surveillance network, from February 2019 to May 2019.

Management of a child with primary ciliary dyskinesia.

Primary ciliary dyskinesia (PCD) is an autosomal recessive condition characterized by dysmotile cilia. Typically associated with defects in the cilia structure, it results in impaired mucociliary clearance of pathogens from the lungs and sinuses. Consequently, patients suffer from recurrent sinopulmonary and middle ear infections.

Pragmatic multicentre randomised controlled trial evaluating the impact of a routine molecular point-of-care 'test-and-treat' strategy for influenza in adults hospitalised with acute respiratory illness (FluPOC): trial protocol.

Background: Influenza infections often remain undiagnosed in patients admitted to hospital due to lack of routine testing. When tested for, the diagnosis and treatment of influenza are often delayed due to the slow turnaround times of centralised laboratory PCR testing. Newer molecular systems, have comparable accuracy to laboratory PCR testing, and can generate a result in under 1 hour, making them potentially deployable as point-of-care tests (POCTs).

Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against .

Development of a vaccine to limit the impact of antibiotic resistant is now a global priority. Serum bactericidal antibody (SBA) is a possible indicator of protective immunity to , but conventional assays measure colony forming units (CFU), which is time-consuming. A luminescent assay that quantifies ATP as a surrogate measure of bacterial viability was tested on strains FA1090, MS11 and P9-17 and compared to CFU-based readouts.

Impact of Baseline Tuberculin Skin Test and Isoniazid Chemoprophylaxis on Subsequent Quantiferon-TB Gold In-Tube Performance in Young Children Assessed After Tuberculosis Contact in Catalonia.

We investigated the impact of baseline tuberculin skin tests (TSTs) and preventive isoniazid chemoprophylaxis on subsequent QuantiFERON-TB Gold In-Tube (QFT-GIT) assays performed after a 10- to 12-week window period in 114 children <5 years of age. Previous TSTs and chemoprophylaxis had no impact on the magnitude of subsequent antigen-induced responses in QFT-GIT. Furthermore, previous TSTs did not induce conversion from a negative to a positive QFT-GIT result.

A method for the generation of large numbers of dendritic cells from CD34+ hematopoietic stem cells from cord blood.

Dendritic cells (DCs) play a central role in regulating innate and adaptive immune responses. It is well accepted that their regulatory functions change over the life course. In order to study DCs function during early life it is important to characterize the function of neonatal DCs.

Feasibility of Point-of-Care Testing for Influenza Within a National Primary Care Sentinel Surveillance Network in England: Protocol for a Mixed Methods Study.

Background: Point-of-care testing (POCT) for influenza promises to provide real-time information to influence clinical decision making and improve patient outcomes. Public Health England has published a toolkit to assist implementation of these tests in the UK National Health Service.

Objective: A feasibility study will be undertaken to assess the implementation of influenza POCT in primary care as part of a sentinel surveillance network.

Pollen exposure weakens innate defense against respiratory viruses.

Background: Hundreds of plant species release their pollen into the air every year during early spring. During that period, pollen allergic as well as non-allergic patients frequently present to doctors with severe respiratory tract infections. Our objective was therefore to assess whether pollen may interfere with antiviral immunity.

Validation of pediatric health-related quality of life instruments for primary ciliary dyskinesia (QOL-PCD).

Rationale: Having developed the first disease-specific, health-related quality of life (HRQoL) instruments for children with primary ciliary dyskinesia (PCD), we aimed to assess the psychometric performance of quality of life (QOL)-PCD child, adolescent, and parent-proxy versions in terms of reliability and validity across cross-cultural settings and caring for patients with this rare disease.

Methods: Children (n = 71), adolescents (n = 85), and parents (n = 68) from multiple centers in the UK and North America completed age-appropriate QOL-PCD and generic QOL measures: pediatric QOL inventory, COPD assessment test (CAT), and Sino-Nasal Outcome Test 20. Total of 13 children, 13 parents, and 17 adolescents repeated QOL-PCD 10 to 14 days later to assess test-retest reliability.

Immunological profiling of key inflammatory drivers of nasal polyp formation and growth in chronic rhinosinusitis.

Background: Chronic rhinosinusitis (CRS) is a chronic inflammatory condition of the upper airways, often associated with the formation of nasal polyps (CRSwNP). It is well established that macroscopically normal (non-polypoidal) sinonasal mucosa in CRSwNP patients can undergo polypoidal change over time, turning into frank polyps. However, little is known about what drives this process.

Impact of point-of-care testing for respiratory viruses on antibiotic use in adults with exacerbation of airways disease.

Background: The ResPOC study demonstrated that syndromic molecular point-of-care testing (POCT) for respiratory viruses was associated with early discontinuation of unnecessary antibiotics compared to routine clinical care. Subgroup analysis suggests these changes occur predominantly in patients with exacerbation of airways disease. Use of molecular POCT for respiratory viruses is becoming widespread but there is a lack of evidence to inform the choice between multiplex syndromic panels versus POCT for influenza only.

Use of Human Fallopian Tube Organ in Culture (FTOC) and Primary Fallopian Tube Epithelial Cells (FTEC) to Study the Biology of Neisseria gonorrhoeae Infection.

Epithelial cells represent one of the most important physical barriers to many bacterial pathogens. In the case of Neisseria gonorrhoeae, the epithelial cell response is critical because they are the main target of the tissue damage triggered by the pathogen, particularly when the organism reaches the Fallopian tube (FT). Although the irreversible damage triggered by N.

Basic Methods for Examining Neisseria gonorrhoeae Interactions with Host Cells In Vitro.

The obligate human pathogen Neisseria gonorrhoeae colonizes primarily the mucosal columnar epithelium of the male urethra and the female endocervix. In addition, gonococci can infect the anorectal, pharyngeal, and gingival mucosae and epithelial cells of the conjunctiva. More rarely, the organism can disseminate through the bloodstream, which can involve interactions with other host cell types, including blood vessel endothelial cells and innate immune cells such as dendritic cells, macrophages, and neutrophils.

Cloning, Expression, and Purification of Recombinant Neisseria gonorrhoeae Proteins.

Modern DNA recombinant techniques and major advances in genetic engineering have resulted in the development of bacterial expression systems that guarantee an unlimited supply of valuable proteins that have potential clinical or industrial use, but which are often limited by their low natural availability. This chapter provides the reader with a general scheme to clone, express, and purify native histidine (His)-tagged proteins in the desired quantity and quality required for its intended use, and reviews the most important factors affecting the production of recombinant proteins in a soluble form. Alternative methods for purification of insoluble recombinant proteins under denaturing conditions are also discussed.

Preparation of Lipooligosaccharide (LOS) from Neisseria gonorrhoeae.

Neisseria gonorrhoeae is a gram-negative obligate human pathogen that contains lipooligosaccharide (LOS) as a major constituent within the outer membrane. LOS plays a major role in pathogenesis by inducing host inflammatory responses and also enabling evasion of host innate immunity through sialylation. Epitopes within LOS are also potential vaccine candidates.

Gonococcal Colony Typing.

Gonococcal colony typing is part a science and part an art that has been central to studies which have identified crucial virulence antigens and also demonstrated the ability of the bacteria to undergo rapid phase and antigenic variation. Without this fundamental work, modern molecular biological studies of gonococcal pathogenesis would not have been possible. Indeed colony typing is still essential when performing biological experiments with clinical and laboratory isolates and for monitoring their outcome.