Development and characterisation of a decellularised bovine osteochondral biomaterial for cartilage repair.

It is proposed that an acellular natural osteochondral scaffold will provide a successful repair material for the early intervention treatment of cartilage lesions, to prevent or slow the progression of cartilage deterioration to osteoarthritis. Here, we investigated the efficacy of methods for the decellularisation of bovine osteochondral plugs. The plugs were subject to four freeze/thaw cycles followed by two cycles of washes in hypotonic solution and low concentration (0.1 % w/v) sodium dodecyl sulphate with protease inhibitors. Plugs were treated with nuclease (DNase and RNase) treatment followed by sterilization in peracetic acid. Full tissue decellularisation was achieved as confirmed by histological analysis and DNA quantification, however the resultant acellular matrix had reduced glycosaminoglycan content which led to an increased percent deformation of cartilage. Furthermore, the acellular scaffold was not reproducibly biocompatible. Additional terminal washes were included in the process to improve biocompatibility, however, this led to visible structural damage to the cartilage. This damage was found to be minimised by reducing the cut edge to cartilage area ratio through decellularisation of larger cuts of osteochondral tissue

Distribution and Population Dynamics of Plant-Parasitic Nematodes in Oregon Vineyards, and Their Effects on Vine Growth [1995-1996]

Objectives 1. Survey Oregon vineyards for the presence, identity and abundance of plant-parasitic nematodes. 2. Relate the distribution and abundance of potentially pathogenic species to viticultural practices and site characteristics. 3. Evaluate the efficacy of Nemacur for reducing populations of plant-parasitic nematodes. 4. Document seasonal changes in the abundance of potentially pathogenic species of plant-parasitic nematodes in order to identify optimum times for sampling

Comparison of Meldola’s Blue Staining and Hatching Assay with Potato Root Diffusate for Assessment of Globodera sp. Egg Viability

Laboratory-based methods to test egg viability include staining with Meldola's Blue and/or juvenile (J2) hatching assays using potato root diffusate (PRD). These two methods have not been tested under identical conditions to directly compare their assessments of Globodera egg viability. Using two bioassay strategies, cysts from a Glabodera sp. population found in Oregon were subjected to both viability assessment methods. In strategy one, intact cysts were first stained with Meldola's Blue (primary staining) and eggs were then transferred to PRD (secondary hatching). In the second strategy, intact cysts were exposed to PRD (primary hatching) and then unhatched eggs were transferred to Meldola's Blue (secondary staining). Two different cohorts of cysts were evaluated using these experimental strategies: cohort I was comprised Of cysts produced on potato in the greenhouse that exhibited low hatch when exposed to PRD and cohort 2 consisted of field-collected cysts whose eggs yielded significant hatch when exposed to PRD Percentage viability was calculated and is expressed as the number of hatched J2 or unstained eggs/total number of eggs within a cyst. With field-produced cysts, primary staining with Meldola's Blue and hatching with PRD produced similar viability estimates, with averages of 74.9% and 76.3%, respectively. In contrast, with greenhouse-produced cysts the two methods yielded much lower and unequal estimates 32.4% to 2.2%, respectively for primary hatching and staining methods. In addition, J2 hatch from Unstained (viable) greenhouse-produced eggs was 13.7% after secondary exposure to PRD compared to 61.5% for field-produced eggs. The majority of eggs remaining unhatched after primary exposure to PRD (> 87%) stained with Meldola's Blue regardless of cyst cohort. Staining with Meldola's Blue provided a conservative assessment of egg viability compared to hatch assay with PRD regardless of diapause.Keywords: egg hatch, juvenile, Globodera, Meldola’s Blue, method, potato root diffusate, cyst nematod

Distribution and Population Dynamics of Plant-Parasitic Nematodes in Oregon Vineyards, and Their Effects on Vine Growth [1993-1994]

Objectives: 1. Survey Oregon vineyards for the presence, identity and abundance of plant parasitic nematodes. 2. Document seasonal changes in abundance of species of important plant parasites in order to identify optimum times for sampling. 3. Evaluate the efficacy of Nemacur for reducing populations of plant parasitic nematodes

Distribution and Population Dynamics of Plant-Parasitic Nematodes in Oregon Vineyards, and Their Effects on Vine Growth [1994-1995]

Objectives 1. Survey Oregon vineyards for the presence, identity and abundance of plant-parasitic nematodes. 2. Relate the distribution and abundance of potentially pathogenic species to viticultural practices and site characteristics. 3. Evaluate the efficacy of Nemacur for reducing populations of plant-parasitic nematodes. 4. Document seasonal changes in the abundance of potentially pathogenic species of plant-parasitic nematodes in order to identify optimum times for sampling

A large scale hearing loss screen reveals an extensive unexplored genetic landscape for auditory dysfunction

The developmental and physiological complexity of the auditory system is likely reflected in the underlying set of genes involved in auditory function. In humans, over 150 non-syndromic loci have been identified, and there are more than 400 human genetic syndromes with a hearing loss component. Over 100 non-syndromic hearing loss genes have been identified in mouse and human, but we remain ignorant of the full extent of the genetic landscape involved in auditory dysfunction. As part of the International Mouse Phenotyping Consortium, we undertook a hearing loss screen in a cohort of 3006 mouse knockout strains. In total, we identify 67 candidate hearing loss genes. We detect known hearing loss genes, but the vast majority, 52, of the candidate genes were novel. Our analysis reveals a large and unexplored genetic landscape involved with auditory function

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2–4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Determinants of recovery from post-COVID-19 dyspnoea: analysis of UK prospective cohorts of hospitalised COVID-19 patients and community-based controls

Background The risk factors for recovery from COVID-19 dyspnoea are poorly understood. We investigated determinants of recovery from dyspnoea in adults with COVID-19 and compared these to determinants of recovery from non-COVID-19 dyspnoea. Methods We used data from two prospective cohort studies: PHOSP-COVID (patients hospitalised between March 2020 and April 2021 with COVID-19) and COVIDENCE UK (community cohort studied over the same time period). PHOSP-COVID data were collected during hospitalisation and at 5-month and 1-year follow-up visits. COVIDENCE UK data were obtained through baseline and monthly online questionnaires. Dyspnoea was measured in both cohorts with the Medical Research Council Dyspnoea Scale. We used multivariable logistic regression to identify determinants associated with a reduction in dyspnoea between 5-month and 1-year follow-up. Findings We included 990 PHOSP-COVID and 3309 COVIDENCE UK participants. We observed higher odds of improvement between 5-month and 1-year follow-up among PHOSP-COVID participants who were younger (odds ratio 1.02 per year, 95% CI 1.01–1.03), male (1.54, 1.16–2.04), neither obese nor severely obese (1.82, 1.06–3.13 and 4.19, 2.14–8.19, respectively), had no pre-existing anxiety or depression (1.56, 1.09–2.22) or cardiovascular disease (1.33, 1.00–1.79), and shorter hospital admission (1.01 per day, 1.00–1.02). Similar associations were found in those recovering from non-COVID-19 dyspnoea, excluding age (and length of hospital admission). Interpretation Factors associated with dyspnoea recovery at 1-year post-discharge among patients hospitalised with COVID-19 were similar to those among community controls without COVID-19. Funding PHOSP-COVID is supported by a grant from the MRC-UK Research and Innovation and the Department of Health and Social Care through the National Institute for Health Research (NIHR) rapid response panel to tackle COVID-19. The views expressed in the publication are those of the author(s) and not necessarily those of the National Health Service (NHS), the NIHR or the Department of Health and Social Care. COVIDENCE UK is supported by the UK Research and Innovation, the National Institute for Health Research, and Barts Charity. The views expressed are those of the authors and not necessarily those of the funders

Genetic mechanisms of critical illness in COVID-19.

Host-mediated lung inflammation is present1, and drives mortality2, in the critical illness caused by coronavirus disease 2019 (COVID-19). Host genetic variants associated with critical illness may identify mechanistic targets for therapeutic development3. Here we report the results of the GenOMICC (Genetics Of Mortality In Critical Care) genome-wide association study in 2,244 critically ill patients with COVID-19 from 208 UK intensive care units. We have identified and replicated the following new genome-wide significant associations: on chromosome 12q24.13 (rs10735079, P = 1.65 × 10-8) in a gene cluster that encodes antiviral restriction enzyme activators (OAS1, OAS2 and OAS3); on chromosome 19p13.2 (rs74956615, P = 2.3 × 10-8) near the gene that encodes tyrosine kinase 2 (TYK2); on chromosome 19p13.3 (rs2109069, P = 3.98 ×  10-12) within the gene that encodes dipeptidyl peptidase 9 (DPP9); and on chromosome 21q22.1 (rs2236757, P = 4.99 × 10-8) in the interferon receptor gene IFNAR2. We identified potential targets for repurposing of licensed medications: using Mendelian randomization, we found evidence that low expression of IFNAR2, or high expression of TYK2, are associated with life-threatening disease; and transcriptome-wide association in lung tissue revealed that high expression of the monocyte-macrophage chemotactic receptor CCR2 is associated with severe COVID-19. Our results identify robust genetic signals relating to key host antiviral defence mechanisms and mediators of inflammatory organ damage in COVID-19. Both mechanisms may be amenable to targeted treatment with existing drugs. However, large-scale randomized clinical trials will be essential before any change to clinical practice