PCR examination of bronchoalveolar lavage samples is a useful tool in pre-clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte).

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung tumour of sheep caused by Jaagsiekte sheep retrovirus (JSRV). The disease is a particular problem in flocks in many parts of the world. The aim of the study was to assess screening methods for individual animals as a prelude to future eradication trials. Results of histological examination were used as the standard to evaluate the relative sensitivity and specificity of an established heminested polymerase chain reaction (PCR) test for JSRV proviral DNA from blood and bronchoalveolar lavage (BAL) samples. PCR results from tissue samples are included as control data. PCR testing of blood samples was found to have an estimated sensitivity of only 10% (95% confidence interval (CI) 3-20) while the sensitivity of the PCR test on BAL samples was 89% (CI 79-96) in comparison to the results of histological examination. We conclude that PCR testing of BAL samples is an effective confirmatory test for sheep with suspected clinical OPA. It is also a useful tool for the pre-clinical identification of individual infected sheep within an infected flock and therefore may prove beneficial in future control or eradication programmes.

Eradication of ovine pulmonary adenocarcinoma by motherless rearing of lambs.

The principles of maedi-visna eradication programmes were applied to a field trial for the eradication of ovine pulmonary adenocarcinoma (OPA). In two maternal flocks the prevalence of gross and histological lesions in slaughtered animals was 18.3 per cent and 29.8 per cent, respectively. The lambing period was supervised for three consecutive years from 1999 to 2001, during which the lambs were taken away from their mothers at birth, deprived of maternal colostrum, and hand-reared away from other sheep. Over the three-year period, 322 hand-reared animals, mainly male lambs between 10 and 14 months old, were slaughtered; their lungs were examined grossly, 52.5 per cent of them were examined histologically, and 105 samples of caudal mediastinal lymph nodes were examined by PCR. No OPA tumours were detected in the slaughter specimens from the derived flock, but one lamb had histological lesions in one lung location; intrauterine transmission was ruled out in this case. No clinical OPA has subsequently been observed in the hand-reared flock. Bronchoalveolar lavage samples from the breeding stock were examined by PCR in order to rule out further subclinical cases of OPA. No Jaagsiekte retrovirus was detected in any of the 488 samples.

Lymphocytotoxic antibody detection and crossmatching for renal transplantation.

Sera of 134 recipients awaiting first renal transplants have been tested for cytotoxic antibody by three methods in order to assess the different techniques, and various factors in the techniques have been investigated. The use of only one technique would have failed to detect some of the antibodies found in the sera of 35 patients and it was necessary to use both a 37 C test and the NIH 20 C technique in order to demonstrate them. Results on 402 serial samples from 108 patients indicate the importance, because of fluctuation in antibody levels. of using more than one sample, i.e., previous sample in addition to a fresh specimen from any patient when crossmatching prior to renal transplantation. Lymphocyte suspensions from 44 of 118 cadaver donors were found to be unsatisfactory for accurate cytotoxic crossmatching due to a high background of dead cells. DNase renders such suspensions suitable for use and should be used in such cases. It is felt that if more than one technique were generally used, improved detection of cytotoxic activity would result and would help to resolve present conflicting opinions about the effects of cytotoxic antibody on graft survival. Results are given of 19 patients who had formed cytotoxic antibody prior to renal transplantation in the Unit. Eighteen of these have surviving grafts.

A new finding relating to transfusion and renal transplants.

Results in 29 recipients of second renal transplants from cadaver donors show a significantly better graft survival at 1 year of 90% in 10 recipients who had not received blood transfusion before their first transplant compared to 41% in 19 recipients who had been transfused prior to their first transplant (P = 0.025).

Immunosuppression with polyunsaturated fatty acids in renal transplantation.

A double-blind controlled trial has been undertaken to assess the value of a preparation containing polyunsaturated fatty acids (PUFA) in human cadaveric renal transplantation. Eighty-nine patients were studied and followed for 6 months after transplantation. Forty-four took the PUFA preparation and 45 the placebo (oleic acid). Other immunosuppression was standardised. Functional graft survival was significantly better in the PUFA group than in those taking the placebo during the first 3 to 4 months post-transplant. At 6 months, however, although the difference between the groups persisted, it was no longer statistically significant. Complications were equally distributed between the groups.

Faecal elastase 1: a marker of exocrine pancreatic insufficiency in cystic fibrosis.

Pancreatic elastase 1 (E1), a digestive protease, is synthesized by the acinar cells of the pancreas. Using an enzyme-linked immunosorbent assay, we evaluated stool E1 levels in the following groups of patients. (a) Specimens submitted for occult blood examination from 20 adults, over 3 consecutive days, to assess the inter-day variability in E1 excretion. There were no symptoms suggestive of pancreatic insufficiency in this group. The mean E1 concentration over all samples was 457 micrograms E1/g stool (range 124-1683). The intra-assay variation was 6.4% (n = 14) and the inter-assay variation was 8.8% (n = 12). The mean intra-patient variation was 17%. (b) Cystic fibrosis (CF) patients. Eight patients had E1 levels in the reference range (> 200 micrograms E1/g stool). The remaining 25 patients had undetectable E1 levels. (c) A control group of children presenting with unexplained bronchiectasis and/or recurrent respiratory infections and no symptoms of pancreatic dysfunction. The mean E1 concentration in the group was 519 micrograms E1/g stool (range 139-1941). There was no significant difference in E1 concentrations between the two non-CF groups, nor between the pancreatic-sufficient CF patients when compared with both non-CF groups. There was a significant difference between the pancreatic-sufficient and -insufficient CF groups (P < 0.001) using the Mann Whitney U test. All fifteen CF patients who were delta F508 homozygotes had undetectable E1. It may be possible to relate CF genotype to the presence or absence of E1 and to the degree of pancreatic insufficiency. Measurement of faecal E1 in children with CF appears to differentiate them into a group of children with normal pancreatic function and a larger group with severe insufficiency.

Lack of a specific immune response against a recombinant capsid protein of Jaagsiekte sheep retrovirus in sheep and goats naturally affected by enzootic nasal tumour or sheep pulmonary adenomatosis.

Enzootic nasal tumour (ENT) and sheep pulmonary adenomatosis (SPA) are two contagious adenocarcinomas of the respiratory tract of sheep and goats. Both diseases are associated with related, but distinct, type-D-retroviruses (ENTV and JSRV respectively). No evidence of circulating antibodies has been described in animals affected by either ENT or SPA using antigens from natural sources. We evaluated the usefulness of a recombinant JSRV capsid protein (JSRV-CA) as antigen to study the antibody responses of animals naturally affected by ENT or SPA, using immunoblotting. Positive reactions were detected in the sera of both affected and unaffected sheep and goats. The reactivity was abolished completely by absorption with the GST fusion partner but not by JSRV-CA, suggesting that it was not specific. The results support prior observations indicating that sheep and goats infected by JSRV and ENTV do not develop specific humoral responses to these retroviruses.

Formation of anti-N-like antibodies in dialysis patients: effect of different methods of dialyzer rinsing to remove formaldehyde.

Use of formalin to sterilize dialyzers is known to be responsible for the formation of anti-N-like antibody in long-term hemodialysis patients. Patients dialyzed as in-patients using formalin were found to be completely free of anti-N-like antibody, while among those on home dialysis, there was a high prevalence (31%) and incidence. The hospital patients were found to be receiving concentrations of formaldehyde less than 1 microgram/ml while those on home dialysis received 3-13 micrograms/ml. This is offered as an explanation for the absence of anti-N-antibody in patients using formalin-sterilized dialyzers.

Pneumonia of lambs following inoculation of isolates of sheep herpesvirus (caprine herpesvirus 1) of different DNA genotypes.

One-day-old, specific pathogen-free lambs, were equally susceptible to infection with three isolates of caprine herpesvirus 1 (CHV1). One of these isolates was genomically different by DNA analysis. Lesions, which were confined to the lung, ranged from a mild interstitial reaction to widespread consolidation. CHV1 was recovered from lungs and less commonly from liver and adrenal gland. Three-week-old SPF lambs were also successfully infected with CHV1. Histopathological findings were similar to those in 1-week-old lambs.

Bile acid and lysolecithin concentrations in the stomach in patients with duodenal ulcer before operation and after treatment by highly selective vagotomy, partial gastrectomy, or truncal vagotomy and drainage.

Duodenogastric reflux of bile acids and lysolecithin in the course of a standard test meal was measured in normal people and in patients with duodenal ulcer before operation and more than one year after highly selective vagotomy, Polya partial gastrectomy, truncal vagotomy and pyloroplasty, and truncal vagotomy and gastrojejunostomy. Before operation, duodenal ulcer patients had significantly higher fasting, post-prandial, and peak bile acid concentrations in the stomach than had normal subjects. After Polya partial gastrectomy, fasting, post-prandial, and peak concentrations of bile acids and lysolecithin were significantly higher than in preoperative duodenal ulcer patients. After highly selective vagotomy, in contrast, bile acid concentrations in the stomach were significantly lower than in preoperative duodenal ulcer patients and post-prandial and peak lysolecithin concentrations were less than half (NS) those recorded in preoperative duodenal ulcer patients. After highly selective vagotomy, bile acid concentrations were also significantly lower than bile acid concentrations after Polya partial gastrectomy, truncal vagotomy and pyloroplasty, and truncal vagotomy and gastrojejunostomy; and post-prandial and peak lysolecithin concentrations were significantly lower than after Polya partial gastrectomy and truncal vagotomy and gastrojejunostomy. Thus, when used in the treatment of patients with duodenal ulcer, highly selective vagotomy keeps ;bile' out of the stomach, probably through its effect on gastric smooth muscle, combined with the preservation of an intact antropyloroduodenal segment. In contrast, Polya partial gastrectomy, truncal vagotomy and gastrojejunostomy, and truncal vagotomy and pyloroplasty all lead to a significant increase in reflux of bile acids and lysolecithin into the stomach. The clinical importance of these findings is that both gastritis and, in the long term, gastric carcinoma may prove to be less common after highly selective vagotomy than after partial gastrectomy or vagotomy with a drainage procedure.

HLA-DR antigens in primary biliary cirrhosis: lack of association.

A study of HLA-DR antigen in 75 patients with primary biliary cirrhosis has been carried out in order to test the hypothesis that genetic factors related to genes controlling immune responses might be important in the pathogenesis of primary biliary cirrhosis. The frequencies of HLA-DR locus antigens was not significantly different from those in 200 normal controls, nor were those of tissue antigens on the A and B loci. No HLA-DR antigen was significantly associated with the appearance of granulomata on liver biopsy (possibly good prognosis) or with raised serum bilirubin (possibly bad prognosis); nor was there any association between any HLA-DR antigen and adverse reactions to D-penicillamine treatment in 17 patients with such adverse reactions. It is concluded that genetic traits related to HLA antigens studied are probably not important in the aetiology of the disease.

HLA antigens and spinal muscular atrophy.

Fifty-four patients with spinal muscular atrophy were examined and HLA typing performed on blood from each. A, B and DR antigens were determined. No statistical difference in antigen prevalence was found between patients and a control population. The clinical and genetic heterogeneity found in this disorder does not appear to be related to HLA status.

Bile reflux and degree of gastritis in patients with gastric ulcer: before and after operation.

The incidence and degree of bile reflux and gastritis has been measured in normal subjects and in patients with gastric ulcer before operation and after treatment by highly selective vagotomy with ulcer excision, Billroth 1 partial gastrectomy, and truncal vagotomy and drainage. Before operation patients had significantly higher (P less than 0.001) bile acid concentrations in the stomach than normal subjects. Treatment by highly selective vagotomy resulted in significantly lower bile acid concentrations than those before operation and those found after Billroth 1 partial gastrectomy. Antral and body gastritis was significantly less in normal subjects than in the preoperative and all postoperative groups. There was no significant difference in antral or body gastritis between the preoperative gastric ulcer patients and the patients after any of the surgical procedures despite the significant differences in bile acids. Though highly selective vagotomy in the treatment of gastric ulcer results in a reduction in duodenogastric reflux of bile there is no improvement in the gastritis that is present.

Enzootic nasal tumour of goats: demonstration of a type D-related retrovirus in nasal fluids and tumours.

Nasal exudate and tumour tissue from goats with enzootic nasal tumours were shown to contain a reverse transcriptase activity associated with a particle of buoyant density typical of retroviruses. The same particle contained a 25,000 Mr protein that cross-reacted with the p27 of Mason-Pfizer monkey virus (MPMV) and with p25 of sheep pulmonary adenomatosis retrovirus. It also contained a low Mr protein related to p10-12 of MPMV.

Antisera with Ia specificity selected by lymphocytes from patients with chronic lymphocytic leukemia.

Cells from patients with chronic lymphocytic leukaemia (CLL) have been used to screen the sera of pregnant women for antibodies specifc for B lymphocytes. The sera have been divided into groups showing Ig specificity: group I (UK 3) had a positive association with HLA-B8 and DW3, and group II (UK 2) with HLA-B7 and DW2. Groups III, IV and X were independent of each other and of HLA-A, B and C locus antigens. Certain sera with B-cell activity showed specific blocking activity in the mixed lymphocyte reaction (MLR), and this activity was dependent on the Ia sepcificities and DW specificities of cells used in the MLR. Some sera specifically blocked the responding cells, other the stimulating cells.