The experiences of NHS hospital acute medicine departments in England during the first wave of the COVID-19 pandemic.

We conducted a survey exploring the experiences of NHS hospital acute medicine services in England during the 1st wave of the COVID-19 pandemic. Responses were collected from 26th May to 8th July 2020. The results of 91 sites are presented. The total number of patients referred to the medical take for assessment and admitted from the medical take decreased from pre-pandemic levels compared to peak COVID-19 activity. The total number of acute medical beds decreased, however critical care beds increased by 162%. We report the median timeline from first admission of COVID-19 to when baseline critical care capacity was reached. We found regional variation across the results. These findings can assist healthcare leaders prepare for future pandemics.

Evolution of asexual and sexual reproduction in the aspergilli.

Aspergillus nidulans has long-been used as a model organism to gain insights into the genetic basis of asexual and sexual developmental processes both in other members of the genus Aspergillus, and filamentous fungi in general. Paradigms have been established concerning the regulatory mechanisms of conidial development. However, recent studies have shown considerable genome divergence in the fungal kingdom, questioning the general applicability of findings from Aspergillus, and certain longstanding evolutionary theories have been questioned. The phylogenetic distribution of key regulatory elements of asexual reproduction in A. nidulans was investigated in a broad taxonomic range of fungi. This revealed that some proteins were well conserved in the Pezizomycotina (e.g. AbaA, FlbA, FluG, NsdD, MedA, and some velvet proteins), suggesting similar developmental roles. However, other elements (e.g. BrlA) had a more restricted distribution solely in the Eurotiomycetes, and it appears that the genetic control of sporulation seems to be more complex in the aspergilli than in some other taxonomic groups of the Pezizomycotina. The evolution of the velvet protein family is discussed based on the history of expansion and contraction events in the early divergent fungi. Heterologous expression of the A. nidulans abaA gene in Monascus ruber failed to induce development of complete conidiophores as seen in the aspergilli, but did result in increased conidial production. The absence of many components of the asexual developmental pathway from members of the Saccharomycotina supports the hypothesis that differences in the complexity of their spore formation is due in part to the increased diversity of the sporulation machinery evident in the Pezizomycotina. Investigations were also made into the evolution of sex and sexuality in the aspergilli. MAT loci were identified from the heterothallic Aspergillus (Emericella) heterothallicus and Aspergillus (Neosartorya) fennelliae and the homothallic Aspergillus pseudoglaucus (=Eurotium repens). A consistent architecture of the MAT locus was seen in these and other heterothallic aspergilli whereas much variation was seen in the arrangement of MAT loci in homothallic aspergilli. This suggested that it is most likely that the common ancestor of the aspergilli exhibited a heterothallic breeding system. Finally, the supposed prevalence of asexuality in the aspergilli was examined. Investigations were made using A. clavatus as a representative 'asexual' species. It was possible to induce a sexual cycle in A. clavatus given the correct MAT1-1 and MAT1-2 partners and environmental conditions, with recombination confirmed utilising molecular markers. This indicated that sexual reproduction might be possible in many supposedly asexual aspergilli and beyond, providing general insights into the nature of asexuality in fungi.

HLA-A, -B, -DR allele group frequencies in 7007 kidney transplant list patients in 27 UK centres.

This observational study aims to determine the HLA specificity frequencies of patients on the UK renal transplant list, which can be used as a resource for those laboratories that support the UK renal transplant programme. Whilst the HLA specificity frequencies may differ from that of the general population, it is the individuals on the transplant list who are in need of a new kidney, which has to be provided from the general population. Any differences in protein allele frequencies between this patient population and the general population are likely to be minimal because of the very large number of patients included. The HLA-A, -B and -DR allele group frequencies from 7007 patients on the UK kidney transplant list (August, 2009) were analysed. HLA types had been submitted to NHSBT to register patients on the UK deceased donor kidney waiting list. The data were submitted from 27 different registering centres throughout the UK. Within this data set, 25 different HLA-A, 50 HLA-B and 18 HLA-DR allele groups were present. The most common allele groups at each locus were -A2 (phenotype frequency 42.6%), -B44 (phenotype frequency 23.3%) and -DR4 (phenotype frequency 29.8%). The least common allele groups at each locus were -A19, - A43, -B16, -B21, -B22, -B83 and -DR5. Reports of HLA frequency (protein allotype) data from populations as large as this are not readily available adding value to this observational study.

Exercise stress test utility in patients with chest pain presumed to be of cardiac origin.

INTRODUCTION: NICE stated exercise stress tests (EST) should not be used to diagnose obstructive coronary artery disease in patients presenting with chest pain presumed to be of cardiac origin. METHODS: A retrospective review of 209 patients with presumed cardiac chest pain was done. EST results, GRACE scores and need for invasive coronary angiogram (ICA) were analysed to predict the need for readmission, intervention and future events. RESULTS: The sensitivity of the EST in identifying obstructive coronary artery disease was 70%. The EST, ICA and the GRACE 6-month mortality had a 77%, 70% and 81% negative predictive value (NPV) for readmission respectively. CONCLUSION: EST, GRACE scores and ICA are useful in providing prognostic information but are poor predictors of readmission. Follow up and education programmes are needed to reduce this burden.

Speciation despite globally overlapping distributions in Penicillium chrysogenum: the population genetics of Alexander Fleming's lucky fungus.

Eighty years ago, Alexander Fleming described the antibiotic effects of a fungus that had contaminated his bacterial culture, kick starting the antimicrobial revolution. The fungus was later ascribed to a putatively globally distributed asexual species, Penicillium chrysogenum. Recently, the species has been shown to be genetically diverse, and possess mating-type genes. Here, phylogenetic and population genetic analyses show that this apparently ubiquitous fungus is actually composed of at least two genetically distinct species with only slight differences detected in physiology. We found each species in air and dust samples collected in and around St Mary's Hospital where Fleming worked. Genotyping of 30 markers across the genome showed that preserved fungal material from Fleming's laboratory was nearly identical to derived strains currently in culture collections and in the same distinct species as a wild progenitor strain of current penicillin producing industrial strains rather than the type species P. chrysogenum. Global samples of the two most common species were found to possess mating-type genes in a near 1:1 ratio, and show evidence of recombination with little geographic population subdivision evident. However, no hybridization was detected between the species despite an estimated time of divergence of less than 1MYA. Growth studies showed significant interspecific inhibition by P. chrysogenum of the other common species, suggesting that competition may facilitate species maintenance despite globally overlapping distributions. Results highlight under-recognized diversity even among the best-known fungal groups and the potential for speciation despite overlapping distribution.

Joint British Diabetes Societies guideline for the management of diabetic ketoacidosis.

The Joint British Diabetes Societies guidelines for the management of diabetic ketoacidosis (these do not cover Hyperosmolar Hyperglycaemic Syndrome) are available in full at: (i) http://www.diabetes.org.uk/About_us/Our_Views/Care_recommendations/The-Management-of-Diabetic-Ketoacidosis-in-Adults; (ii) http://www.diabetes.nhs.uk/publications_and_resources/reports_and_guidance; (iii) http://www.diabetologists-abcd.org.uk/JBDS_DKA_Management.pdf. This article summarizes the main changes from previous guidelines and discusses the rationale for the new recommendations. The key points are: Monitoring of the response to treatment (i) The method of choice for monitoring the response to treatment is bedside measurement of capillary blood ketones using a ketone meter. (ii) If blood ketone measurement is not available, venous pH and bicarbonate should be used in conjunction with bedside blood glucose monitoring to assess treatment response. (iii) Venous blood should be used rather than arterial (unless respiratory problems dictate otherwise) in blood gas analysers. (iv) Intermittent laboratory confirmation of pH, bicarbonate and electrolytes only. Insulin administration (i) Insulin should be infused intravenously at a weight-based fixed rate until the ketosis has resolved. (ii) When the blood glucose falls below 14 mmol/l, 10% glucose should be added to allow the fixed-rate insulin to be continued. (iii) If already taking, long-acting insulin analogues such as insulin glargine (Lantus(®), Sanofi Aventis, Guildford, Surry, UK) or insulin detemir (Levemir(®), Novo Nordisk, Crawley, West Sussex, UK.) should be continued in usual doses. Delivery of care (i) The diabetes specialist team should be involved as soon as possible. (ii) Patients should be nursed in areas where staff are experienced in the management of ketoacidosis.

Redefining genera of cereal pathogens: Oculimacula, Rhynchosporium and Spermospora.

The taxonomy of Oculimacula, Rhynchosporium and Spermospora is re-evaluated, along with that of phylogenetically related genera. Isolates are identified using comparisons of DNA sequences of the internal transcribed spacer ribosomal RNA locus (ITS), partial translation elongation factor 1-alpha (tef1), actin (act), DNA-directed RNA polymerase II largest (rpb1) and second largest subunit (rpb2) genes, and the nuclear ribosomal large subunit (LSU), combined with their morphological characteristics. Oculimacula is restricted to two species, O. acuformis and O. yallundae, with O. aestiva placed in Cyphellophora, and O. anguioides accommodated in a new genus, Helgardiomyces. Rhynchosporium s. str. is restricted to species with 1-septate conidia and hooked apical beaks, while Rhynchobrunnera is introduced for species with 1-3-septate, straight conidia, lacking any apical beak. Rhynchosporium graminicola is proposed to replace the name R. commune applied to the barley scald pathogen based on nomenclatural priority. Spermospora is shown to be paraphyletic, representing Spermospora (type: S. subulata), with three new species, S. arrhenatheri, S. loliiphila and S. zeae, and Neospermospora gen. nov. (type: N. avenae). Ypsilina (type: Y. graminea), is shown to be monophyletic, but appears to be of minor importance on cereals. Finally, Vanderaaea gen. nov. (type: V. ammophilae), is introduced as a new coelomycetous fungus occurring on dead leaves of Ammophila arenaria. Citation: Crous PW, Braun U, McDonald BA, Lennox CL, Edwards J, Mann RC, Zaveri A, Linde CC, Dyer PS, Groenewald JZ (2020). Redefining genera of cereal pathogens: Oculimacula, Rhynchosporium and Spermospora. Fungal Systematics and Evolution 7: 67-98. doi: 10.3114/fuse.2021.07.04.

British Society for Histocompatibility & Immunogenetics and British Transplantation Society guidelines for the detection and characterisation of clinically relevant antibodies in allotransplantation.

Ongoing technological developments in antibody detection and characterisation allowing relative quantitation of HLA-specific antibody levels, combined with crossmatch results, now allow a graded assessment of patient potential donor immunological risk for allotransplantation, rather than a simple 'positive' or 'negative' categorization of crossmatch results. These developments have driven a thorough revision of the British Society for Histocompatibility & Immunogenetics and British Transplantation Society Guidelines for the Detection and Characterisation of Clinically Relevant Antibodies in Allotransplantation. These newly published revised Guidelines contain a number of recommendations as to best practice for antibody detection and crossmatching for the transplantation of a wide range of solid organs and tissues. These recommendations are briefly summarized in this article.

Keratin synthesis during development of the embryonic chick feather.

The synthesis of keratin proteins during development of the embryonic chick feather was studied by quantitative gel electrophoresis of the reduced and carboxymethylated proteins. The results demonstrated a coordinated synthesis of the major keratin proteins, during and after the onset of keratin synthesis. The results from gel electrophoresis correlated well with electron microscope visualization or keratin fibrils in the developing feathers. Autoradiography at the electron microscope level indicated that the feather cells lose the ability to synthesize DNA before keratin synthesis begins, but retain the ability to synthesize RNA after keratin synthesis begins.

UV Spectrum and Proposed Role of Diethylberyllium in a 7Li-7Be Solar Neutrino Experiment.

Although measurement of the solar neutrino flux via the (7)Li(v(e),e(-))(7)Be reaction was proposed many years ago, no experiment has been implemented since it has been difficult to identify a sensitive (7)Be detection technique. Here it is proposed that the (7)Be atom be incorporated into a volatile molecule, placed in a buffer-gas-filled cell, and then extracted by photodissociation; after excitation by a tunable laser, bursts of photons would be detected. The absorption spectrum of the molecular candidate diethylberyllium has been measured between 186 and 270 nanometers in a spectrophotometer to determine the required photodissociation laser wavelength and intensity.

Inscription of fibre Bragg gratings in non-sensitised fibres using VUV F2 laser radiation.

We report the inscription of fibre Bragg gratings in non-sensitised SMF 28 and HI 980 fibre by exposure to VUV F2 laser radiation at 157 nm. The modulated effective refractive index change Deltan(eff) deduced from the shift in the grating reflection peaks was Deltan(eff) = 2.8x10(-4) and 1.7x10(-4) in SMF 28 and HI 980 fibre respectively. The possible influence of non-uniformity of core exposure and VUV cladding absorption loss on these results is discussed.

The Role of Catalyst Support, Diluent and Co-Catalyst in Chromium-Mediated Heterogeneous Ethylene Trimerisation.

Sequential treatment of a previously-calcined solid oxide support (i.e. SiO2, γ-Al2O3, or mixed SiO2-Al2O3) with solutions of Cr{N(SiMe3)2}3 (0.71 wt% Cr) and a Lewis acidic alkyl aluminium-based co-catalyst (15 molar equivalents) affords initiator systems active for the oligomerisation and/or polymerisation of ethylene. The influence of the oxide support, calcination temperature, co-catalyst, and reaction diluent on both the productivity and selectivity of the immobilised chromium initiator systems have been investigated, with the best performing combination (SiO2-600, modified methyl aluminoxane-12 {MMAO-12}, heptane) producing a mixture of hexenes (61 wt%; 79% 1-hexene), and polyethylene (16 wt%) with an activity of 2403 g gCr -1 h-1. The observed product distribution is rationalised by two competing processes: trimerisation via a supported metallacycle-based mechanism and polymerisation through a classical Cossee-Arlman chain-growth pathway. This is supported by the indirect observation of two distinct chromium environments at the surface of the oxide support by a solid-state 29Si NMR spectroscopic study of the Cr{N(SiMe3)2}x/SiO2-600 pro-initiator.

Two-photon excitation studies of m-THPC photosensitizer and photodynamic activity in an epithelial cell line.

A femtosecond laser delivering pulses of wavelength 800nm and 124fs duration at rates of 1kHz has been used to investigate the two-photon excited fluorescence in the photosensitizer m-THPC. The scaling of fluorescence amplitude with laser power and fluorescence sidelight imaging are found to support a predominantly two-photon excitation mechanism. A value for the two-photon cross-section of δ=1.8×10(-57)m(4)s is derived by comparing the fluorescence signals excited by wavelengths of 800 and 400nm. Preliminary results demonstrating the two-photon induced PDT activity of m-THPC in an epithelial cell line are also reported.

Sexual and vegetative compatibility genes in the aspergilli.

Gene flow within populations can occur by sexual and/or parasexual means. Analyses of experimental and in silico work are presented relevant to possible gene flow within the aspergilli. First, the discovery of mating-type (MAT) genes within certain species of Aspergillus is described. The implications for self-fertility, sexuality in supposedly asexual species and possible uses as phylogenetic markers are discussed. Second, the results of data mining for heterokaryon incompatibility (het) and programmed cell death (PCD) related genes in the genomes of two heterokaryon incompatible isolates of the asexual species Aspergillus niger are reported. Het-genes regulate the formation of anastomoses and heterokaryons, may protect resources and prevent the spread of infectious genetic elements. Depending on the het locus involved, hetero-allelism is not tolerated and fusion of genetically different individuals leads to growth inhibition or cell death. The high natural level of heterokaryon incompatibility in A. niger blocks parasexual analysis of the het-genes involved, but in silico experiments in the sequenced genomes allow us to identify putative het-genes. Homologous sequences to known het- and PCD-genes were compared between different sexual and asexual species including different Aspergillus species, Sordariales and the yeast Saccharomyces cerevisiae. Both het- and PCD-genes were well conserved in A. niger. However some point mutations and other small differences between the het-genes in the two A. niger isolates examined may hint to functions in heterokaryon incompatibility reactions.

Objective determination of claw pain and its relationship to limb locomotion score in dairy cattle.

We hypothesized that claw and foot pain could be objectively determined and have a strong effect on limb locomotion. Claw pain was measured using hoof testers equipped with a pressure gauge. Soft tissue pain was evaluated with an algometer. Pain was determined as the maximum pressure recorded at the time the limb was withdrawn following claw or soft tissue compression with the hoof tester or algometer. Locomotion scores and claw and soft tissue pain were determined on 263 Holstein cows from 2 commercial dairy farms. The frequency and the magnitude of pain had an effect on locomotion score in the ipsilateral limb for lateral, but not medial, claws. The magnitude of the lateral claw pain index for limbs with locomotion scores 1 to 5 was 0.95 +/- 0.01, 0.90 +/- 0.02, 0.67 +/- 0.04, 0.65 +/- 0.05, and 0.45 +/- 0.11, respectively. The magnitude of the medial claw pain index for limbs with locomotion scores 1 to 5 was 1.0 +/- 0.00, 0.99 +/- 0.01, 0.98 +/- 0.01, 1.0 +/- 0.00, and 1.0 +/- 0.00, respectively. The frequency of painful claws (n = 208) in limbs with locomotion scores 1, 2, and > or =3 was 0.529, 0.173, and 0.298, respectively. The frequency of painless claws (n = 318) in limbs with locomotion scores 1, 2, or > or =3 was 0.792, 0.088, and 0.120, respectively. The frequency of pain (27.1%) and total lesions (85.6%) was greater in lateral claws (n = 524) than that of pain (2.1%) and total lesions (14.4%) in medial claws (n = 524). Yet the magnitude of the pain index in sore claws was similar for medial (0.73 +/- 0.09) and lateral claws (0.64 +/- 0.02). The magnitude and frequency of claw pain in one hind limb was inconsistently and weakly affected by locomotion score or claw pain in the contralateral limb. The prevalence of unilateral (32.8%) and bilateral (23.3%) pain was similar and lower than the occurrence of bilaterally nonpainful claws (43.9%) in the study group. Painful claws (n = 78) occurred on sound limbs (n = 332) with a pain index (0.72 +/- 0.02) indicative of less pain than the pain index (0.61 +/- 0.02) of painful claws (n = 130) on lame limbs (n = 192). The results showed that lateral claw pain was related to ipsilateral limb locomotion score and subclinical pain was a relatively common occurrence. Objective measures of pain may provide a more reliable, continuous measure of clinical events used in modeling lameness.

The role of the Pirani scoring system in the management of club foot by the Ponseti method.

The Pirani scoring system, together with the Ponseti method of club foot management, was assessed for its predictive value. The data on 70 idiopathic club feet successfully treated by the Ponseti method and scored by Pirani's system between February 2002 and May 2004 were analysed. There was a significant positive correlation between the initial Pirani score and number of casts required to correct the deformity. A foot scoring 4 or more is likely to require at least four casts, and one scoring less than 4 will require three or fewer. A foot with a hindfoot score of 2.5 or 3 has a 72% chance of requiring a tenotomy. The Pirani scoring system is reliable, quick, and easy to use, and provides a good forecast about the likely treatment for an individual foot but a low score does not exclude the possibility that a tenotomy may be required.

In Vitro Chromatin Assembly: Strategies and Quality Control.

Chromatin accessibility is modulated by structural transitions that provide timely access to the genetic and epigenetic information during many essential nuclear processes. These transitions are orchestrated by regulatory proteins that coordinate intricate structural modifications and signaling pathways. In vitro reconstituted chromatin samples from defined components are instrumental in defining the mechanistic details of such processes. The bottleneck to appropriate in vitro analysis is the production of high quality, and quality-controlled, chromatin substrates. In this chapter, we describe methods for in vitro chromatin reconstitution and quality control. We highlight the strengths and weaknesses of various approaches and emphasize quality control steps that ensure reconstitution of a bona fide homogenous chromatin preparation. This is essential for optimal reproducibility and reliability of ensuing experiments using chromatin substrates.

Donor CD31 genotype and its association with acute graft-versus-host disease in HLA identical sibling stem cell transplantation.

CD31 gene polymorphisms are implicated in the pathogenesis of graft-versus-host disease (GvHD) following haematopoietic stem cell transplantation (HST). We investigated the influence of CD31 genotype on the incidence of GvHD following HST from an human leukocyte antigen (HLA)-identical sibling donor. Donor and recipient CD31 codons 125, 563 and 670 DNA polymorphisms were determined in 85 cases of HLA identical sibling HST from two transplant centres. A correlation between CD31 genotype and acute GvHD was considered significant if observed in patients from both transplant centres independently. A strong correlation was identified between donor CD31 codon 125 genotype and the incidence of acute GvHD. Acute GvHD grades II-IV occurred in 27 of 46 (59%) recipients with a CD31 codon 125 leucine / valine heterozygous donor compared to nine of 39 (23%) recipients with a CD31 codon 125 homozygous donor (P=0.0019, relative-risk 2.45, 95% confidence interval 1.3-4.5). This correlation was significant in patients from both transplant centres (P=0.015 and P=0.019). We suggest that CD31 genotype may influence the function of donor-derived leukocytes and may be informative when there is a choice of comparable donors.