Antiviral antibodies in rheumatoid arthritis discordant HLA-identical same-sexed sibling pairs.
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Biomedical subjects
Publications and source records attributed to C R Madeley.
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We have developed a nucleic acid dot-blot hybridization test for the detection of astroviruses in stool samples. The test was not as sensitive as electron microscopy for the detection of low numbers of well preserved astrovirus particles, but was able to identify astroviruses in stools containing particles of indistinct morphology. In total, this procedure identified astroviruses in more samples than did electron microscopy, and the data indicate that the incidence of astroviruses may be substantially underestimated.
We report conditions for the growth of human faecal astrovirus in a continuous colonic carcinoma cell line (CaCo-2). Purified particles contained three polypeptides, one of which (24k) appeared loosely held on the exterior.
Recent evidence has implicated adenovirus 12 in the aetiology of coeliac disease so that persistent infection by this virus must be considered. We have undertaken a search for adenovirus DNA in duodenal biopsy samples from a total of 26 coeliac and non-coeliac patients. We could find no evidence of persistent virus DNA by Southern blot techniques even under conditions which approach a sensitivity of one copy of virus genome per cell, and use either adenovirus 12 or 41 DNA.
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Adenovirus DNA was isolated directly from virus-containing stools and digested with restriction endonucleases. The resulting fragments were separated by polyacrylamide gel electrophoresis (PAGE) and visualized by silver staining. This enabled us to assign most of the viruses detected to subgenus, serotype and, sometimes, unique strains. Although less sensitive than electron microscopy, the method allowed more information about the infecting virus to be obtained and no cultivation was necessary. Comparison with culture also allowed dual infections to be recognized. A 2-year survey of faecal adenoviruses in Newcastle upon Tyne showed that type 41 (strain 41a) was the predominant type and strain 41p was not recorded. Heterogeneity in strain 41a was also noted as found elsewhere. Adenovirus type 40 was common prior to 1985 but was absent during the last 2 years.
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Mouse monoclonal antibodies, directed against antigenic sites on influenza A and B viruses and found to be type-specific in an immunoassay, were assessed for use as diagnostic reagents in an indirect immunofluorescence assay on nasopharyngeal secretions. The influenza A antibodies were directed against nucleoprotein or matrix protein antigens and the influenza B antibodies against nucleoprotein and haemagglutinin antigens. The influenza A anti-matrix monoclonal antibody was found to give a strong intranuclear particulate fluorescence in normal baboon kidney cells and cells from nasopharyngeal secretions negative for influenza A virus, including those from a patient infected with respiratory syncytial virus. Pools of the remaining monoclonal antibodies gave satisfactory results on 25 specimens from patients with influenza A H1N1 and H3N2 subtypes and 12 from patients with influenza B.
The relation between diarrhoea and vomiting, the excretion of stool micro-organisms, and the passive intestinal permeability in 20 infants living in a deprived urban area was studied prospectively from birth to age 6 months. Intestinal permeability was measured from the ratio of lactulose to mannitol recovered in the urine of infants receiving feeds containing both markers. Micro-organism excretion was found to occur in both the presence and absence of gastrointestinal symptoms, but a significantly higher mean intestinal permeability was recorded in those infants with symptoms and organisms in the stool than in those with neither. An increased intestinal permeability may be a sign of mucosal damage by intestinal micro-organisms.
A commercially available latex agglutination test, Rotalex (Orion Diagnostics, Finland), for detecting rotaviruses was evaluated in comparison with four other tests (electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay) routinely used in our laboratories. Although Rotalex was the least complex method, it showed lack of specificity and sensitivity when carried out according to the manufacturer's instructions. Four basic modifications of Rotalex are described. These include the use of Hank's balanced salt solution, increasing the incubation time to 20 min, reading the agglutination result by an experienced observer, and the use of 50 mm square glass plates. The modified procedure gave results which were comparable with those obtained by electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. The latter techniques, when used to detect rotavirus, all gave similar results.
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The RNA genome of rotaviruses consists of 11 segments in four size-classes which can be separated by polyacrylamide gel electrophoresis, although 11 separate bands are not shown by all strains. We propose a cryptogram (Rotacode) based on the relative distance of migration of adjacent bands in each size-class for coding the typical pattern of each strain of virus. This provides a shorthand for recording details of each strain and for grouping electrophoretically similar strains. Rotacode was found to be reliable and reproducible, with identical codes being obtained for the same samples in repeated experiments under code and by various observers. Rotacode was also used to analyse 189 strains obtained over a three-year period and differentiated 13 electrophoretypes. This confirms the considerable electrophoretic variability of wild strains.
A simple and rapid procedure for identifying adenovirus and rotavirus in stool extracts has been developed. The technique is based on polyacrylamide gel electrophoresis of the virus nucleic acid, but sample preparation is straightforward and does not entail phenol extraction or the use of a radioactive label. Furthermore, processing is not influenced by the amount of specimen obtained and is thus suitable for application as a batch testing method. This approach removes the need for specific antisera, which are not readily available since most of these viruses cannot be grown using routine tissue culture procedures. Trials in this laboratory have indicated that the technique is of comparable sensitivity to electron microscopy.
Although viruses have recently been implicated in the pathogenesis of Paget's disease and Perthes' disease, little information is available regarding bone and marrow response to the presence of a noncarcinogenic virus. Vaccinia virus was introduced into the medullary cavity of the femoral shaft of 30 New Zealand White rabbits, and an appropriate control inoculation was performed. Virus was recovered from the inoculation site in 50% of the animals at intervals to one month after inoculation. Histologic abnormalities were observed in 69% of the test femora; similar abnormalities were not observed in the control femora. These abnormalities consisted of bone and marrow necrosis, lymphocyte and plasma cell infiltration, and new bone formation. Some of the newly formed bone was also necrotic. The latter observation suggests an alternative mechanism for the death of new bone, which, in Perthes' disease, has customarily been attributed to repeated ischemic insults.
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Attempts were made to isolate viruses from babies' stools that contained adenoviruses detected by electron microscopy. One hundred and fifty-nine specimens from 71 children were studied and adenoviruses of established serotypes were isolated from 81 stools. Serial stool samples containing adenovirus particles were obtained from 35 children, and prolonged shedding of recognized serotypes was common. Simultaneous and sequential infections by different serotypes were also observed. Thirty-six children shed adenoviruses that could not be isolated using cell cultures normally used to detect adenoviruses, and nine of these children also shed adenoviruses of established serotypes. Passage in Chang conjunctival cell culture allowed characterization of fastidious adenoviruses from 14 children as members of a previously unrecognized serotype.
The events occurring during the time in hospital of 24 babies were recorded in detail, particularly those related to bowel function. The babies were admitted to a general paediatric unit with various diagnoses a total of 27 times during a six month period. The purpose of the study was to fill gaps in the records of a previous study (Scott et al. 1979) and to try to relate excretion of viruses detectable by electron microscopy to disturbances of bowel function (diarrhoea and/or vomiting). The results showed that a variety of viruses were associated with gastroenteropathy, that virus excretion could occur without disease and that hospital acquisition of virus is not uncommon. Antiobiotic treatment did not appear to precipitate diarrhoea in this small number of babies.