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Biomedical subjects

P Stevenson

Publications and source records attributed to P Stevenson.

At least 19 recordsLinked to original sources

Application of a monoclonal antibody-based antigen detection enzyme-linked immunosorbent assay (antigen ELISA) for field diagnosis of bovine trypanosomiasis at Nguruman, Kenya.

A monoclonal antibody-based, enzyme immunoassay (antigen ELISA) for the detection of species-specific invariant antigens of Trypanosoma congolense, T. vivax or T. brucei in the serum of infected animals was evaluated as a means of diagnosis using bovine field sera from a trypanosomiasis endemic area, Nguruman, Kenya. Circulating trypanosome antigens were detected in 126 (96.2%) of 131 serum samples from animals with parasitologically confirmed diagnosis: 74.8% were positive for antigens of two or three trypanosome species, while 21.4% tested positive for one trypanosome species. When 70 sera from animals (at Nguruman), which had tested negative for trypanosomes by the buffy coat technique, were tested, 35 (50.0%) of them were shown to be antigen-ELISA positive: 24 (34.3%) showing infection with a single species and 11 (15.7%) with mixed infections. The predominant trypanosome species diagnosed in the two herds by antigen ELISA was T. vivax, which was detected in 133 (82.6%) of the 161 sera that tested positive for antigens, followed by T. congolense in 122 (75.8%) sera, with 109 (67.7%) showing evidence of mixed infections with two or three trypanosome species. In single infections, T. vivax exceeded T. congolense by a ratio of 2:1, with T. brucei accounting for less than 1.0%. Evidence for the specificity of the test was provided by analysis of field sera from 100 cattle, from a trypanosomiasis-free area, infected with other haemoparasites (anaplasmosis, babesiosis and theileriosis), which all tested negative in the assay.

Animals

Common antigenic domains in transferrin-binding protein 2 of Neisseria meningitidis, Neisseria gonorrhoeae, and Haemophilus influenzae type b.

There is now considerable evidence to show that in the Neisseria and Haemophilus species, membrane receptors specific for either transferrin or lactoferrin are involved in the acquisition of iron from these glycoproteins. In Neisseria meningitidis, the transferrin receptor appears to consist of two proteins, one of which (TBP 1) has an M(r) of 95,000 and the other of which (TBP 2) has an M(r) ranging from 68,000 to 85,000, depending on the strain; TBP 2 binds transferrin after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting, but TBP 1 does not do so. The relative contributions of these two proteins to the binding reaction observed with intact cells and to iron uptake are presently unknown. However, they are being considered as potential components of a group B meningococcal vaccine. Analogous higher- and lower-molecular-weight proteins associated with transferrin binding have been found in N. gonorrhoeae and Haemophilus influenzae. Previous work with polyclonal antibodies raised in mice with whole cells of iron-restricted N. meningitidis showed that the meningococcal TBP 2 exhibits considerable antigenic heterogeneity. Here, we report that antiserum against purified TBP 2 from one strain of N. meningitidis cross-reacts on immunoblotting with the TBP 2 of all meningococcal isolates examined, as well as with the TBP 2 of N. gonorrhoeae. This antiserum also cross-reacted with the TBP 2 of several strains of H. influenzae type b, thus showing the presence of common antigenic domains among these functionally equivalent proteins in different pathogens; no cross-reaction was detected with a purified sample of the human transferrin receptor.

Animals

Control of Fasciola gigantica with triclabendazole in Indonesian cattle.

The efficacy of triclabendazole (TCBZ) against natural infections with Fasciola gigantica was evaluated in a field study using 102 Indonesian cattle. The animals were divided into two groups; one received TCBZ at 12 mg/kg orally every eight weeks for one year while the other remained untreated. Relative to controls the Fasciola faecal egg count in treated animals was significantly reduced (P less than 0.01) to almost zero at all observations. There were no differences between the groups in blood values and body mass gain. Plasma glutamate dehydrogenase (GLDH) levels in treated animals were reduced to within normal values, whereas those in controls remained above normal throughout. Although plasma gamma glutamyl transferase (GGT) levels remained within the normal range in both groups they were consistently higher in control than in treated animals. However, both GLDH and GGT levels in control animals were considerably lower than those reported by others in experimental low-level chronic fasciolosis, which suggests that only mild liver damage was caused by small numbers of F. gigantica in these animals. In addition to considerations of disease prevalence and economic data from abattoirs, control strategies for fasciolosis in Indonesian cattle require an evaluation of losses due to subclinical disease.

Animals

Use of ivermectin to control sarcoptic mange in goats in Indonesia.

The results of three experiments aimed at controlling sarcoptic mange in goats in Indonesia are reported. In Experiment 1 treatment with ivermectin (0.2 mg/kg body weight) or coumaphos (0.05% dip solution) resulted in a significant reduction in mite numbers and an increase in body weight while in the untreated controls all animals had to be withdrawn from the experiment due to severity of infection. In Experiment 2 three treatment levels of ivermectin were shown to be equally effective in reducing mite numbers, while again all untreated control animals were withdrawn. The third experiment demonstrated a significantly greater reduction in mite numbers following two doses of ivermectin at a seven day interval when compared to a single dose. Again the majority of untreated control animals were withdrawn. Future control strategies and reasons for failure to eliminate the presence of mites are discussed.

Animals

Homidium bromide as a chemoprophylactic for cattle trypanosomiasis in Kenya.

Homidium bromide was used in a strategic chemoprophylactic regime to control trypanosomiasis in Boran cattle in Kenya. Trypanosome infection rates in cattle receiving homidium bromide prophylaxis were compared with those in control cattle which received no prophylaxis but were treated with diminazene aceturate when infected. Homidium bromide was administered twice during the year after which no infections were detected for periods of nineteen weeks and seventeen weeks respectively. The drug sensitivity of the infecting trypanosomes is believed to be a major factor in determining the duration of prophylaxis.

Animals

Antigenic and molecular heterogeneity of the transferrin-binding protein of Neisseria meningitidis.

The transferrin-binding protein in 35 Neisseria meningitidis isolates was examined using a binding assay involving 125I-transferrin. The results show that most strains have a binding protein with a Mr between 78 kDa and 83 kDa; only 4 strains had a binding protein with a Mr of about 68 kDa. The side of the protein appears unrelated to the serogroup or serotype of the organism. Using antibodies raised to whole cells of N. meningitidis grown under iron restriction we show also that considerable antigenic heterogeneity exists amongst the transferrin-binding proteins. This makes it a less than promising vaccine candidate antigen, although conserved antigenic domains are now being sought.

Antigens, Bacterial

Utilization of enterobactin and other exogenous iron sources by Haemophilus influenzae, H. parainfluenzae and H. paraphrophilus.

The ability of Haemophilus influenzae, H. parainfluenzae and H. paraphrophilus to utilize iron complexes, iron-proteins and exogenous microbial siderophores was evaluated. In a plate bioassay, all three species used not only ferric nitrate but also the iron chelates ferric citrate, ferric nitrilotriacetate and ferric 2,3-dihydroxybenzoate. Each Haemophilus species examined also used haemin, haemoglobin and haem-albumin as iron sources although only H. influenzae could acquire iron from transferrin or from haemoglobin complexed with haptoglobin. None of the haemophili obtained iron from ferritin or lactoferrin or from the microbial siderophores aerobactin or desferrioxamine B. However, the phenolate siderophore enterobactin supplied iron to both H. parainfluenzae and H. paraphrophilus, and DNA isolated from both organisms hybridized with a DNA probe prepared from the Escherichia coli ferric enterobactin receptor gene fepA. In addition, a monospecific polyclonal antiserum raised against the E. coli 81 kDa ferric enterobactin receptor (FepA) recognized an iron-repressible outer membrane protein (OMP) in H. parainfluenzae of between 80 and 82 kDa (depending on the strain). This anti-FepA serum did not cross-react with any of the OMPs of H. paraphrophilus or H. influenzae. The OMPs of each Haemophilus species were also probed with antisera raised against the 74 kDa Cir or 74 kDa IutA (aerobactin receptor) proteins of E. coli. Apart from one H. parainfluenzae strain (NCTC 10665), in which an OMP of about 80 kDa cross-reacted with the anti-IutA sera, no cross-reactivity was observed between Cir, IutA and the OMPs of H. influenzae, H. parainfluenzae or H. paraphrophilus.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Outer Membrane Proteins

Novel aerobactin receptor in Klebsiella pneumoniae.

Several Klebsiella pneumoniae strains which produced enterochelin but not aerobactin were nevertheless sensitive to cloacin DF13. In contrast, a strain of serotype K1:O1 which produced both siderophores was cloacin-resistant. Loss by mutation of the O1 but not K1 antigen rendered this strain cloacin-sensitive, indicating that the O1 antigen prevented access of cloacin to the cloacin/aerobactin receptor. Unlike the K1:O1 strain, the aerobactin-negative strains failed to hybridize in a colony blot assay with an aerobactin receptor gene probe prepared from pColV-K30. However, antisera raised against the 74 kDa pColV-K30 aerobactin receptor cross-reacted with a 76 kDa outer-membrane protein in each K. pneumoniae strain. In addition to the 76 kDa protein, the K1:O1 strain also produced a strongly cross-reacting 74 kDa protein. To determine whether these aerobactin-negative strains could use aerobactin, mutants unable to synthesize siderophores were isolated. Aerobactin promoted the growth of these mutants in iron-deficient media. The evidence presented suggests that some K. pneumoniae strains produce an aerobactin iron-uptake system without apparent production of aerobactin and which is probably based on a 76 kDa receptor, the gene for which does not hybridize with aerobactin receptor gene encoded on pColV-K30.

Bacterial Outer Membrane Proteins

Comparative effects of two antimycotic agents, ketoconazole and terbinafine on the metabolism of tolbutamide, ethinyloestradiol, cyclosporin and ethoxycoumarin by human liver microsomes in vitro.

Two antimycotic agents, the azole ketoconazole and the allylamine terbinafine, have been examined for their effects on the metabolism of tolbutamide, ethinyloestradiol, cyclosporin and ethoxycoumarin by human liver microsomes (n = 4) in vitro. Ketoconazole caused marked inhibition of all enzyme activities with mean IC50 values (concentration producing 50% inhibition) of 17.9 microM (tolbutamide hydroxylase), 1.9 microM (ethinyloestradiol 2-hydroxylase), 2.0 microM (cyclosporin N-demethylase), 2.1 microM (cyclosporin hydroxylase) and 25 microM (ethoxycoumarin O-deethylase). At 50 microM terbinafine concentration, inhibition was less than 5% for tolbutamide and ethoxycoumarin, approximately 12% for both cyclosporin pathways and 35% for ethinyloestradiol. Terbinafine does not have the same inhibitory potential for cytochrome P-450 isozymes as ketoconazole.

Antifungal Agents

Expression in Escherichia coli K-12 of the 76,000-dalton iron-regulated outer membrane protein of Shigella flexneri confers sensitivity to cloacin DF13 in the absence of Shigella O antigen.

One of the chromosomal segments associated with virulence in Shigella flexneri encodes the production of aerobactin and the synthesis of an iron-regulated 76-kilodalton outer membrane protein believed to be the ferric-aerobactin receptor. However, S. flexneri expressing this putative aerobactin receptor, which is slightly larger than that encoded by pColV, is insensitive to the killing action of cloacin DF13, a bacteriocin which binds to other aerobactin receptor proteins and kills the cells. In this paper we show that the conjugal transfer of DNA encoding the iron-regulated 76-kilodalton protein from S. flexneri to Escherichia coli K-12 conferred cloacin DF13 sensitivity on the recipients. However, E. coli K-12 which had also inherited genes specifying Shigella O-antigen biosynthesis remained cloacin insensitive. The data suggest that it is unwise to use cloacin DF13 sensitivity alone to screen transconjugants or clinical isolates for the expression of aerobactin receptor proteins.

Antigens, Bacterial

Leg pain: an uncommon presentation of perforated diverticular disease.

Five cases of perforated diverticular disease are described in which pain in the thigh or leg was the predominant symptom. In four patients pyrexia or leucocytosis were present and three patients ultimately developed surgical emphysema in the left thigh. In retroperitoneal perforation of the colon, the symptoms in the leg may overshadow those in the abdomen and lead to delay in diagnosis.

Aged

Iron-regulated outer-membrane proteins of Escherichia coli strains associated with enteric or extraintestinal diseases of man and animals.

The SDS-PAGE patterns of the iron-regulated outer-membrane proteins from 70 strains of Escherichia coli isolated from various human and animal infections were analysed and the nature of the siderophores produced was examined. Iron-regulated 81 kDa and 74 kDa protein bands seen in SDS-PAGE gels were characterized further by immunoblotting using anti-81 kDa and anti-74 kDa (Cir) sera. The results showed considerable differences between the patterns of the iron-regulated outer-membrane proteins exhibited by the different strains. Nevertheless, three distinct and characteristic profiles, based on the most prominent bands expressed, could be identified, although not all strains produced patterns which matched with one of these. These results suggest the possibility of using the pattern of iron-regulated outer-membrane proteins expressed, as well as siderophores produced, as a new set of markers to characterize groups of pathogenic E. coli.

Animals

Carbamazepine-induced severe systemic hypersensitivity reaction with eosinophilia.

Skin rash, fever, and eosinophilia developed in a previously healthy 35-year-old woman three weeks after starting carbamazepine. Fulminant respiratory and renal failure ensued. Autopsy showed pneumonitis, nephritis, serositis, pancreatitis, hepatitis, and carditis, characterized by an infiltrate of eosinophils and lymphocytes. The severity, duration, and extensive organ involvement of the reaction make this case unique.

Adult

Iron regulated outer membrane proteins of Escherichia coli: variations in expression due to the chelator used to restrict the availability of iron.

Iron restriction was induced in Escherichia coli O 111, E. coli O 164 and E. coli C by growing the organisms in trypticase soy broth containing ovotransferrin, desferal, EDDA (ethylenediamine-dihydroxyphenylacetic acid) or alpha,alpha'-dipyridyl. There were marked qualitative and quantitative differences in the iron regulated outer membrane proteins expressed in the presence of the various iron chelators. Differences in the kinetics of growth were also noted. E. coli C was devoid of a ferric enterobactin iron uptake system.

2,2'-Dipyridyl

Naturally occurring antibodies in human sera that react with the iron-regulated outer membrane proteins of Escherichia coli.

Sera from normal healthy human adults and infants, as well as sera from mice, rabbits, and guinea pigs, were examined by immunoblotting for naturally occurring antibodies reacting with outer membrane proteins of two Escherichia coli strains, O111 and O18. Some individuals had antibodies reacting very strongly with the iron-regulated outer membrane proteins, including the ferric-enterochelin receptor protein (Mr, 81,000), as well as with ompA. However, sera from infants contained predominantly antibodies to ompA; antibodies recognizing the iron-regulated outer membrane proteins were either absent or barely detectable. In human serum the antibodies were mainly of the immunoglobulin G class. No serotype-specific antibodies to the lipopolysaccharide of E. coli O111 or O18 were found in the sera tested.

Antibodies, Bacterial

Synthesis of aerobactin and a 76,000-dalton iron-regulated outer membrane protein by Escherichia coli K-12-Shigella flexneri hybrids and by enteroinvasive strains of Escherichia coli.

One of the chromosomal segments associated with the virulence of Shigella flexneri and transferred to Escherichia coli K-12 by conjugation has been shown to code for the production of aerobactin and for the synthesis of an iron-regulated 76,000-dalton (76K) outer membrane protein. Analysis of various E. coli K-12-S. flexneri transconjugants showed that the genes involved with the synthesis of aerobactin and with the production of the 76K protein were linked to the mtl region of the S. flexneri chromosome. S. flexneri itself synthesized a 76K protein in its outer membrane under iron restriction as well as traces of 81K and 74K proteins. An examination of four enteroinvasive strains of E. coli showed that each produced aerobactin and a 76K outer membrane protein during iron-restricted growth. The profile of the iron-regulated proteins expressed by the enteroinvasive strains of E. coli was virtually identical to that expressed by the laboratory-constructed E. coli K-12-S. flexneri hybrids under the same growth conditions.

Animals

Toxocara and ascaris infection in British pigs: a serological survey.

A serological survey of growing pigs from farms throughout Great Britain revealed that antibodies to toxocara and ascaris larval antigens were present in 4.5 per cent and 39.3 per cent respectively. Heavy infection with toxocara occurred only rarely and it is considered that this parasite is unlikely to be an important cause of chronic focal interstitial hepatitis. Antibodies to both nematodes were detected more frequently in heavy hogs than in pigs of lighter weight. Reginal differences in the prevalence of antibodies were also observed.

Animals