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Biomedical subjects

R A Payne

Publications and source records attributed to R A Payne.

At least 19 recordsLinked to original sources

The prevalence of Toxoplasma infection among pregnant women in Ibadan, Nigeria.

The seroprevalence of toxoplasmosis in pregnant women from the inner area of Ibadan was determined by the dye test. Two hundred and seventy-three of the 352 women (78%) had dye test titres of 1/16 or greater with 165 (47%) having titres of 1/128 or greater. In contrast, 42 of 192 pregnant women (22%) from the Swansea area of the UK were dye test positive with only six (3%) having titres of 1/128. The possibility that reinfection or recrudescence is responsible for maintaining high antibody levels in African women and the consequence of this high level of infection in relation to potential human immune deficiency virus (HIV) infections are discussed. Social and environmental conditions indicate that the source of infection is contact with cat faeces.

Animals

Potential role of IgG avidity for diagnosing toxoplasmosis.

Sera from 20 cases of toxoplasmic lymphadenopathy were examined by an enzyme linked immunosorbent assay toxoplasma IgG avidity (ELISA) at two laboratories. The results obtained were largely in agreement and showed that sera from patients with acute infection had low avidity IgG (30% or less), whereas sera from patients with chronic infection had high avidity IgG (40% or more). It is suggested that this type of assay could have a useful complementary role in antenatal testing for toxoplasmosis.

Acute Disease

Five commercial enzyme linked immunosorbent assay kits for toxoplasma specific IgM antibody.

Five commercially available enzyme linked immunosorbent assay (ELISA) kits for the detection of specific IgM against Toxoplasma gondii were evaluated in a three centre study and results compared with those of the Public Health Laboratory Service ELISA for Toxoplasma IgM (PHL IgM ELISA). Fifty selected sera were tested by all the methods (Toxo-M, Captia Toxo-M EIA, Toxo Enz M EIA, Toxonostika IgM EIA, Sopazyme Toxo IgM EIA) at the three reference centres in England and Wales and 177 routine sera by all the methods in one or other of the centres. Ten of the 50 selected sera contained autoimmune antibodies but no specific IgM and 29 had toxoplasma specific IgM detectable by the PHL IgM ELISA. The kits were assessed for their specificity and sensitivity compared with the PHL IgM ELISA, and the percentage coefficient of variation for binding to the solid phases was determined. They were also rated subjectively by the staff performing the assays and an overall impression of each kit was gained by allocating scores of several criteria. There was quite close agreement among the results obtained with all five commercial assays and the PHL IgM ELISA, although some of the sera pre-selected as being potentially problematic showed the limitations of some of the assays.

Animals

Gastric cancer: a continuing diagnostic challenge--a district general hospital's experience.

We present the results of 87 consecutively referred patients with malignant gastric tumours who presented to the North Middlesex Hospital between 1980 and 1984, inclusively. The results of this prospective study showed a high incidence of adenocarcinoma situated in the cardia (16 per cent of patients), and antral carcinomas presented more commonly with chronic blood loss. Delay in diagnosis was still a serious problem and weight loss remained an underestimated symptom, especially when combined with other upper gastrointestinal symptomology. The majority of tumours were adenocarcinomas. Six patients had unusual tumours which had special therapeutic and prognostic implications. The results of the above study are compared with the European experience of the management of malignant gastric tumours.

Adenocarcinoma

Enzyme-linked immunosorbent assays for the measurement of specific antibodies in experimentally induced ovine toxoplasmosis.

Tachyzoites of the RH strain of Toxoplasma gondii were inoculated intravenously into sheep following which serum samples were collected at approximately weekly intervals for 9 months. The sera were examined by the toxoplasma dye test and two enzyme-linked immunosorbent assays (ELISA) specifically developed for investigations of ovine toxoplasmosis. One was an antibody class capture assay for the detection of anti-toxoplasma specific IgM, the other an indirect assay which detected anti-toxoplasma IgG. Some of the sheep had antibodies to toxoplasma prior to inoculation but none had specific IgM. Sera collected 17 days after inoculation showed that all had raised specific antibody levels but the only sheep that produced specific antitoxoplasma IgM were those that were initially without any antibody. Specific IgM could be detected in all these particular sheep for at least 1 month after infection and up to 3 months in some. Specific IgG persisted at high levels for at least 3 months and could still be detected at moderate levels for at least 9 months. The ELISA methods described are simple to perform and could clearly distinguish between previous infection and this experimental infection with Toxoplasma gondii.

Animals

Rapid indirect enzyme linked immunosorbent assay (ELISA) for detecting antitoxoplasma IgG: comparison with dye test.

A rapid and simple enzyme linked immunosorbent assay (ELISA) for the detection of specific IgG against Toxoplasma gondii was compared with the dye test on 533 serum samples. In general, results were comparable but not with sera that contained high concentrations of toxoplasma specific IgM or that had been heated at 56 degrees C. There were no false positive results with sera containing rheumatoid factor or anti-nuclear factor. It is concluded that if a dye test is not to be performed then the serum should be tested for both toxoplasma specific IgG and IgM to avoid misleading results. Heat inactivated serum should also not be tested in this type of specific IgG assay.

Animals

Public Health Laboratory Service enzyme linked immunosorbent assay for detecting Toxoplasma specific IgM antibody.

An enzyme linked immunosorbent assay (ELISA) based on the antibody class capture method for the detection of specific IgM against Toxoplasma gondii, using the microtitre plate format, was developed. Antigen binding was detected using a monoclonal antibody, CIE3, conjugated to horseradish peroxidase. Prior mixing of the conjugate and antigen improved the stability of these reagents as well as removing an incubation stage from the assay. The incubation time of less than four hours permits a rapid throughput of specimens. Using the assay, a total of 163 sera were examined in a three centre study and good agreement was found. Results were expressed as arbitrary enzyme immunoassay units (EIUs) against a freeze dried standard. Throughout the study the standard serum showed a coefficient of variation less than 10% across the microtitre plate. By measuring IgM titres in patients having toxoplasmic lymphadenopathy with a known date of onset, IgM class antibodies were shown to peak at two months, persisting for about six months. In addition, a case of laboratory acquired toxoplasmosis was monitored. Sera shown to contain rheumatoid factor and antinuclear factor did not give false positive results. This rapid, robust, and simplified assay is used by the Public Health Laboratory Service Toxoplasma Reference Units and will provide a standard with which other assays can be compared.

Animals

Influence of furosemide on rubidium-86 uptake and alpha-adrenergic responsiveness of arterial smooth muscle.

Furosemide-induced inhibition of 86Rb uptake was measured in rat and rabbit aorta and compared with its ability to inhibit contractions induced by alpha-adrenergic agonists. In both rat and rabbit tissues, furosemide defined a portion of 86Rb uptake (IC50 = 2.5 microM) which was distinct from the ouabain-sensitive fraction. Furosemide-sensitive 86Rb uptake was [Cl-]ext dependent and required Na+ and K+ for optimal activity, suggesting that it reflected a Na+-K+ cotransport process. Furosemide-sensitive 86Rb uptake was found to be greater in HEPES buffer than in bicarbonate buffer. Phenylephrine-induced contractions of rat and rabbit aorta were inhibited by furosemide; however, rat responses were far more sensitive. Agonist-induced uptake of 45Ca was reduced by furosemide in rat aorta, but not in rabbit aorta. Agonist-induced 45Ca efflux stimulation was reduced in both species. These findings indicate the presence in arteries of a furosemide-sensitive, Cl-dependent Na+-K+ cotransport process. Along with other monovalent transport processes, it may modulate Ca2+ availability and thereby influence arterial contractility.

Animals

Primary suture or T-tube drainage after choledochotomy.

It has been suggested that decreased morbidity and mortality after choledochotomy is obtained by avoidance of T-tube drainage of the common bile duct. A prospective randomised trial involving 80 patients having either primary suture of the common bile duct or T-tube drainage has therefore been undertaken. The indications for exploring the duct together with the complications arising in the two groups are enumerated. There is a significant saving of hospital time after primary suture, P = 0.05 (Wilcoxon's test).

Adult

Phosphoinositide hydrolysis is correlated with agonist-induced calcium flux and contraction in the rabbit aorta.

In the present study changes in the extent of 32P labelling of membrane phospholipids were correlated with the alpha 1-adrenoceptor-induced events of increased 45Ca influx, 45Ca release and contraction in the rabbit aorta. Under basal conditions 32P incorporation into all phospholipids proceeded without saturation through 80 min of labelling. During a 5 min exposure to 10(-5) M norepinephrine (NE) after 25 min of prelabelling the incorporation of 32P into certain phospholipids was substantially increased. Phosphatidic acid (PA) labelling was increased above basal levels by 4.1 fold, phosphatidylinositol (PI) 2.5 fold and phosphatidylcholine (PC) 1.8 fold. Half maximal stimulation of 32P labelling of PA occurred at 2.0 microM, which was similar to the EC50 value for stimulation of 45Ca influx (2.5 microM) and 45Ca release (2.1 microM) but slightly higher than the value for contractile response (0.9 microM). Antagonist sensitivity studies reinforced the alpha 1 receptor subtype character of the rabbit aorta. Prazosin (10(-7) M) reduced agonist-induced events by 63-82% while yohimbine (10(-7) M) was without influence. Phenoxybenzamine (10(-8) M) reduced agonist-induced events by 56-76%. A temporal comparison showed that agonist stimulation of PA labelling was slower than 45Ca release, but similar to the time course of 45Ca influx. Hydrolysis of 32P-labelled phosphatidylinositol diphosphate (PIP2) was more rapid and paralleled 45Ca release. These findings suggest that PIP2 hydrolysis may account for the rapid phase of norepinephrine-induced contraction in rabbit aorta while PA or its immediate precursor diacylglycerol may account for receptor-induced Ca2+ influx.

Animals

Comparison of a latex agglutination test with other serological tests for the measurement of antibodies to Toxoplasma gondii.

One hundred sera from 49 patients with glandular toxoplasmosis were examined by a latex agglutination test, the dye test, an indirect haemagglutination test, and a double antibody sandwich enzyme linked immunosorbent assay (ELISA) for antitoxoplasma IgM. The results support previous findings that the dye test, indirect haemagglutination test, and latex agglutination test measure different antibodies to Toxoplasma gondii. In early glandular toxoplasmosis, when specific IgM was detected, the titres of both the latex agglutination test and the indirect haemagglutination test were lower than the dye test. Repeat specimens from 11 of the patients showed four cases in which the latex agglutination test titres never exceeded 1/256, whereas both the dye test and the indirect haemagglutination test showed significant titres and specific IgM was detected in every case. We conclude that the latex agglutination test should not be used as a substitute for the dye test in the serological diagnosis of glandular toxoplasmosis. All sera giving a positive latex agglutination test result should be referred for further tests. A combination of the dye test and double antibody sandwich ELISA gives the most reliable serological diagnosis of early glandular toxoplasmosis.

Antibodies

Antibody class capture assay (ACCA) for rubella-specific IgM antibody.

Enzyme-linked immunosorbent assays for IgM antirubella were carried out on 1,546 sera, using an IgM capture method with a F (ab')2 conjugate (ACCA). Under the conditions described, sera containing IgM antirubella bound up to 15 times as much enzyme activity as negative specimens. Paired serum specimens from 27 patients, serial serum specimens from 6 patients, and single serum specimens from 15 patients who had had recent rubella were examined by the haemagglutination inhibition test (HAI) in the presence and absence of 2-mercaptoethanol following sucrose density gradient centrifugation (SDGC). ACCA confirmed all the results found with HAI following SDGC. Specimens were examined from ten patients with congenital rubella; ACCA confirmed the results found with both immunofluorescence following SDGC and radioimmunoassay. Pre- and post-vaccination specimens from 123 patients who had been vaccinated against rubella were examined. An IgM response could only be demonstrated in the 57 cases when IgG was absent in the first specimen. The specificity of the assay was confirmed by testing 31 serum specimens from rubella immune patients that also contained rheumatoid factor, 163 serum specimens from patients with acute infections other than rubella, and 12 serum specimens from infants with miscellaneous neonatal abnormalities other than congenital rubella. The ACCA proved a simple, sensitive, and specific test for IgM antirubella and the results compared favourably with those obtained by the SDGC technique.

Antibodies, Viral

Enzyme-linked immunosorbent assay (ELISA) using antibody class capture for the detection of antitoxoplasma IgM.

Sera from 180 patients with suspected toxoplasmic lymphadenopathy were examined for antitoxoplasma IgM by an enzyme-linked immunosorbent assay (ELISA), using antibody class capture (ACCA). Of 82 positive ACCA results, 78 were confirmed by testing the IgM fractions of the sera, obtained by sucrose density gradient centrifugation (SDGC). The four positive results which could not be confirmed were all from patients with at least a year's history of lymphadenopathy. Sera from 10 patients with low Sabin Feldman dye test (DT) titers gave positive ACCA results and subsequent specimens from them showed a rise in antibody concentration, confirming the diagnosis of acute toxoplasmosis. The antitoxoplasma IgM immunofluorescent antibody test (IgM-IFA) on whole serum was relatively insensitive and gave false-positive results with sera containing rheumatoid factor (RF) and antinuclear factor (ANF). There were no false-positive ACCA results with such sera, probably because the conjugates were prepared from F(ab')2 fragments of antitoxoplasma serum. The ACCA proved to be sensitive, specific and easily automated enabling examination of large numbers of specimens.

Enzyme-Linked Immunosorbent Assay

Cystic tumours of the breast.

A retrospective survey of cystic tumours of the breast seen at the North Middlesex and St Bartholomew's Hospitals, London, has been undertaken to determine their incidence and clinical features. During the years 1967-76 4530 histological examinations were carried out on breast tissue and 1277 breast cancers diagnosed. Intracystic papillary tumours, both benign and malignant, were the least uncommon of cyst tumours, but the malignant type accounted for 0.55% of all the breast cancers. Aspects suggesting the possible presence of an intracystic tumour are discussed.

Adult