Hospitalisation and mortality risk of SARS-COV-2 variant omicron sub-lineage BA.2 compared to BA.1 in England.

The Omicron variant of SARS-CoV-2 became the globally dominant variant in early 2022. A sub-lineage of the Omicron variant (BA.2) was identified in England in January 2022. Here, we investigated hospitalisation and mortality risks of COVID-19 cases with the Omicron sub-lineage BA.2 (n = 258,875) compared to BA.1 (n = 984,337) in a large cohort study in England. We estimated the risk of hospital attendance, hospital admission or death using multivariable stratified proportional hazards regression models. After adjustment for confounders, BA.2 cases had lower or similar risks of death (HR = 0.80, 95% CI 0.71-0.90), hospital admission (HR = 0.88, 95% CI 0.83-0.94) and any hospital attendance (HR = 0.98, 95% CI 0.95-1.01). These findings that the risk of severe outcomes following infection with BA.2 SARS-CoV-2 was slightly lower or equivalent to the BA.1 sub-lineage can inform public health strategies in countries where BA.2 is spreading.

Effect of informational internet web pages on patients' decision-making: randomised controlled trial regarding choice of spinal or general anaesthesia for orthopaedic surgery.

This study explored whether patients' preference for particular types of anaesthesia could be influenced pre-operatively by giving them the addresses of various relevant websites. Patients at an orthopaedic pre-assessment education clinic completed a questionnaire, which included a short multiple-choice general knowledge quiz about anaesthesia, and also questioned them as to their choice of anaesthesia (general or neuraxial). Patients were randomly assigned to intervention or control groups. Intervention group members were given the addresses of three relevant anaesthesia and health related websites to access at home. All patients were asked to complete the questionnaires on a second occasion, before surgery. Initially, most patients stated a preference for general anaesthesia. Subsequently, the intervention group altered their preference towards neuraxial anaesthesia compared to the control group (p < or = 0.0001). The increase in median (IQR [range]) anaesthesia knowledge test score was greater in the intervention group (from 10.0 (9.0-12.0 [5.0-14.0]) to 13.0 (11.0-14.0 [6.0-14.0])) than in the control group (from 10.0 (9.0-11.5 [3.0-13.0]) to 11.0 (9.0-12.0 [4.0-14.0]); p = 0.0068).

The small muscle-specific protein Csl modifies cell shape and promotes myocyte fusion in an insulin-like growth factor 1-dependent manner.

We have isolated a murine cDNA encoding a 9-kD protein, Chisel (Csl), in a screen for transcriptional targets of the cardiac homeodomain factor Nkx2-5. Csl transcripts were detected in atria and ventricles of the heart and in all skeletal muscles and smooth muscles of the stomach and pulmonary veins. Csl protein was distributed throughout the cytoplasm in fetal muscles, although costameric and M-line localization to the muscle cytoskeleton became obvious after further maturation. Targeted disruption of Csl showed no overt muscle phenotype. However, ectopic expression in C2C12 myoblasts induced formation of lamellipodia in which Csl protein became tethered to membrane ruffles. Migration of these cells was retarded in a monolayer wound repair assay. Csl-expressing myoblasts differentiated and fused normally, although in the presence of insulin-like growth factor (IGF)-1 they showed dramatically enhanced fusion, leading to formation of large dysmorphogenic "myosacs." The activities of transcription factors nuclear factor of activated T cells (NFAT) and myocyte enhancer-binding factor (MEF)2, were also enhanced in an IGF-1 signaling-dependent manner. The dynamic cytoskeletal localization of Csl and its dominant effects on cell shape and behavior and transcription factor activity suggest that Csl plays a role in the regulatory network through which muscle cells coordinate their structural and functional states during growth, adaptation, and repair.

Real-time whole genome sequencing to control a Streptococcus pyogenes outbreak at a national orthopaedic hospital.

BACKGROUND: Whole genome sequencing (WGS) of Streptococcus pyogenes linked to invasive disease has been used to identify and investigate outbreaks. The clinical application of WGS in real-time for outbreak control is seldom employed. AIMS: A fatal case of bacteraemia at a national orthopaedic hospital prompted an outbreak investigation to identify carriers and halt transmission using real-time WGS. METHODS: Retrospective surveillance was conducted to identify patients with Streptococcus pyogenes infections in the last year. Upon contact tracing, four patients and 179 staff were screened for Streptococcus pyogenes carriage. All isolates identified were emm-typed. WGS was performed in real-time on a subset of isolates. FINDINGS: Twelve isolates of Streptococcus pyogenes from the index case, two patients and eight staff were identified. Six isolates were emm 1.0, including the index case and five staff isolates. The remaining isolates belonged to distinct emm types. WGS analysis was undertaken on the six emm 1.0 isolates. Five were indistinguishable by single nucleotide polymorphism (SNP) analysis, with 0 SNP distance, and one had one SNP difference, supporting the hypothesis of recent local transmission. All screen-positive healthcare workers were offered treatment with penicillin or clindamycin. No further cases were identified. CONCLUSION: The increased molecular discrimination of WGS confirmed the clustering of these cases and the outbreak was contained. This demonstrates the clinical utility of WGS in managing outbreaks of invasive Streptococcus pyogenes in real-time and we recommend its implementation as a routine clinical service.

Identification of mutations in the WT1 gene in tumours from patients with the WAGR syndrome.

Individuals with constitutional, heterozygous deletions of chromosome region 11p13 are predisposed to the development of Wilms' tumour, indicating the site of the tumour predisposition gene. The WT1 gene is a candidate for this cancer predisposition gene. If this gene is truly involved in tumorigenesis it would be expected to be mutant in tumour tissue from patients with 11p13 deletions. We have used single-stranded conformation polymorphism (SSCP) and polymerase chain reaction sequencing to test this hypothesis in an exon-by-exon analysis of the gene. Four tumours were analysed, two of which were from unilaterally affected individuals and two from a bilaterally affected patient. SSCP analysis identified mutations in the two unilateral tumours which, on sequencing, were shown to involve a 10-bp insertion in exon 7 and a single base pair change in exon 8. Both mutations result in the generation of premature stop codons and the predicted proteins would lack part of the zinc finger motif. Despite complete sequencing of the WT1 gene in both of the bilateral tumours, no mutations were identified. These results possibly suggest that WT1 may not be involved in tumorigenesis in all tumours. All four tumours retained heterozygosity in the 11p15 region, making it unlikely that a second recessive oncogene in this region was involved in tumorigenicity.

Co-expression of the Bordetella pertussis leader peptidase I results in enhanced processing and expression of the pertussis toxin S1 subunit in Escherichia coli.

Bordetella pertussis is the causative agent of whooping cough. Traditional vaccines against this disease are inherently reactogenic, thus research is currently focussed on the production of less reactive, acellular vaccines. Expression of candidate antigens for these vaccines in Escherichia coli would be preferable, however, several B. pertussis antigens undergo incorrect post-translational processing in E. coli. The leader peptidase gene (lep) of B. pertussis encodes a protein of 294 amino acid residues that shares homology with other prokaryote leader peptidase I sequences. Hydrophilicity analysis based on the predicted amino acid sequence has demonstrated a similar membrane topology to that of E. coli and Salmonella typhimurium leader peptidase I. Co-expression of the B. pertussis lep gene in E. coli strain TOPP2 expressing the pertussis toxin S1 subunit was found to markedly increase the expression and post-translational processing of the S1 protein.

Effects of benzodiazepines on laryngeal reflexes. Comparison of lormetazepam and Diazemuls.

Of 20 volunteers, five were given intravenous Diazemuls 15 mg over 15 seconds, and three groups of five were given lormetazepam 2 mg intravenously over 10, 20 and 60 seconds, respectively. Laryngeal reactivity and psychomotor function were tested at intervals from prior to injection until 4 hours after injection. For equivalent degrees of depression of psychomotor function, lormetazepam depressed the laryngeal reflex less than Diazemuls (p = 0.004). Lormetazepam give over 60 seconds depressed the laryngeal reflex more than when given over 10 seconds (p = 0.008) or over 20 seconds (p = 0.048), although a significant difference was not demonstrated between the 10-second and 20-second groups. These results concur with experimental evidence that benzodiazepine receptor multiplicity exists, which allows various members of the benzodiazepine group of drugs to exhibit differing therapeutic ratios for their various effects.

Effects of halothane, enflurane and isoflurane anaesthesia on renal plasma flow.

Effective renal plasma flow (ERPF) and linear cardiac output (aortic blood velocity) were measured in 15 patients who received halothane, enflurane or isoflurane in oxygen. All three agents caused a significant reduction in ERPF (P less than 0.05) and the effect was greater at 1.25 MAC than at 0.75 MAC. No significant difference was demonstrated between the agents. Linear cardiac output did not change significantly during the study, suggesting that the observed reduction in ERPF was not caused by cardiovascular depression.

Loss of heterozygosity at 11p13 in Wilms' tumours does not necessarily involve mutations in the WT1 gene.

Loss of heterozygosity (LOH) in tumour cells is generally accepted as 'exposing' recessive cancer genes. The short arm of chromosome 11 shows consistent LOH in Wilms' tumours along its entire length. Occasionally, however, only the 11p13 and/or the 11p15 regions are involved. Deletions of the 11p13 region consistently predisposes to Wilms' tumorigenesis. We have analysed the recently cloned WT1 gene from the 11p13 region exon-by-exon in five tumours previously shown to have undergone LOH for the 11p13 region, using single strand conformation polymorphism analysis (SSCP) and PCR sequencing. Our analysis using SSCP failed to identify any band shifts in the WT1 gene from these tumours. In addition we also sequenced the zinc finger region of WT1, which is the part of the gene most frequently showing mutations. Only the normal sequence was found in all of these tumours. These results demonstrate that LOH in Wilms' tumours is not always related to mutations in the WT1 genes and argues strongly that another gene, probably in the 11p15 region, may be more important in Wilms' tumorigenesis.

Comparison of the effects of dopamine, dobutamine, and dopexamine upon renal blood flow: a study in normal healthy volunteers.

1. We compared the effects of dopexamine, dopamine and dobutamine on the heart rate, blood pressure and renal blood flow of six healthy volunteers in an open triple crossover trial. 2. The results suggest that at the dose ranges investigated dopamine was the most effective agent for increasing renal blood flow.

A single base pair polymorphism in the WT1 gene detected by single-strand conformation polymorphism analysis.

The Wilms' tumor predisposition gene, WT1, was analysed exon-by-exon in a variety of tumours using the single-strand conformation polymorphism (SSCP) technique. A consistent variation in the usual band pattern for exon 7 was detected in this survey. On sequencing, a silent mutation was noted in codon 313 resulting in an A-->G transition in an arginine codon. The A-->G transition destroys an AflIII restriction enzyme recognition site, which provides a rapid means of identifying heterozygotes at this locus. Analysis of the segregation of this polymorphism in families demonstrated a co-dominant inheritance pattern. In an analysis of 21 randomly selected individuals 25% were heterozygous at this locus, which makes this polymorphism useful in a variety of genetic analyses.

Constitutional mutations in the WT1 gene in patients with Denys-Drash syndrome.

The Denys-Drash syndrome is characterised by a typical nephropathy, genital abnormalities and also predisposes to the development of Wilms' tumor. These patients eventually go into end stage renal failure. A candidate Wilms' tumor gene, WT1, from the 11p13 chromosome region has recently been cloned. We have analysed the DNA sequence in constitutional cells from eight patients and have shown heterozygous mutations in six of them. Four of the mutations were in exon 9, all resulting in missense mutations. Three were at nucleotide position 1180 resulting in an arg > trp amino acid change. The other was at position 1186 converting an asp > asn in the predicted resultant protein. One patient had a missense mutation in exon 8, converting an arg > his. A single base pair insertion at nucleotide position 821 in exon 6 resulted in the generation of a premature stop codon in the last patient. We were unable to find a mutation in one patient despite complete sequencing of the genomic sequence of the gene. The last patient carried a constitutional deletion of the 11p13 region and no additional mutation was found. There was no obvious correlation between the type of mutation and phenotypic expression. These results further demonstrate that the WT1 gene is important in both the development of the kidney and the genito-urinary system.

Comparison of the analgesic efficacy and plasma concentrations of high-dose intra-articular and intramuscular morphine for knee arthroscopy.

BACKGROUND AND OBJECTIVE: It is important to provide good postoperative analgesia after discharge from day case surgery. The usefulness of intra-articular morphine for analgesia after day case knee arthroscopy remains controversial. A large dose of morphine intra-articularly may provide a good long-lasting analgesia, but its efficacy and pharmacokinetics are not known and may be no better than intramuscular morphine. We compared the effect of 10 mg intra-articular and intramuscular morphine for 24 h post-injection in a randomized double-blind study. METHODS: Forty adults undergoing knee arthroscopy were recruited and received either 10mg morphine intra-articularly or intramuscularly. Our primary outcome was overall visual analogue assessment of pain (0-100 mm scale where 0 is no pain and 100 is worst possible pain) between 4 h (on discharge) and 24 h (post-operatively). Plasma morphine concentrations were measured at 15 min, and 1, 2, 4 and 24 h. The use of additional analgesia was noted. RESULTS: The assessment of pain experienced between discharge (4 h) and 24 h was significantly better in the intra-articular (n = 20; mean+/-SD: 18+/-19) than the intramuscular (n = 19; mean+/-SD: 34+/-20) group (P = 0.027). The number of patients consuming any additional analgesia between discharge and 24 h was significantly lower in the intra-articular morphine group (P = 0.038), with 4 (20%) patients in the intra-articular group and 11 (60%) patients in the intramuscular group consuming supplementary analgesia. There were no differences in plasma morphine concentrations between the groups. CONCLUSIONS: A large dose of intra-articular morphine provided better analgesia than the same dose of intramuscular morphine, low plasma morphine levels suggesting a peripheral mechanism.