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Biomedical subjects

J S Gray

Publications and source records attributed to J S Gray.

At least 19 recordsLinked to original sources
PubMed2004

Babesia divergens (Phylum Apicomplexa) in vitro growth in the presence of calf serum.

Resistance to severe babesiosis in young calves has frequently been ascribed to an unknown serum factor(s) which inhibits growth of Babesia bovis in vitro. Our experiments show that young calf sera are as suitable as adult bovine sera for the in vitro culture of Babesia divergens, indicating that in this species at least inverse age resistance is due to alternative mechanisms. The suitability of commercial foetal calf sera for B. divergens cultures seems highly variable.

Animals↗
PubMed2002

Naturally acquired babesiosis in a reindeer (Rangifer tarandus tarandus) herd in Great Britain.

A provisional diagnosis of babesiosis was made in a reindeer herd in Scotland when seven animals died during 1997 and 1998. Additional clinical cases occurred, but the animals recovered after treatment. Thirty-one reindeer from the herd were tested for the prevalence of exposure to Babesia by the indirect fluorescent antibody test using a bovine isolate of Babesia divergens that had been passaged through gerbils. Infection rates were determined by Giemsa-stained blood smears. In addition, molecular identification of the infecting Babesiasp. was undertaken using SSU rRNA gene sequence analysis. It is likely that the organism causing babesiosis in this reindeer herd is B. divergens.

Animals↗
PubMed2002

Invasion, and short- and long-term survival of Babesia divergens (Phylum Apicomplexa) cultures in non-bovine sera and erythrocytes.

In order to explore the feasibility of producing a Babesia divergens live vaccine free of bovine material contaminants the parasite's ability to grow in human, sheep and horse erythrocytes and serum and serum-free medium was investigated. B. divergens was successfully maintained in bovine erythrocytes overlaid with serum-free HL-1 medium. Supplementation of the culture medium with bovine or sheep serum improved parasite growth (monitored by measuring parasitaemia and uptake of tritiated hypoxanthine) whereas horse and human sera reduced parasite growth. As assessed by Giemsa's stained and FITC-labelled blood smears, the parasite invaded all erythrocyte types. Polyparasitism was less common in sheep and horse erythrocytes than in bovine and human erythrocytes. Accole stages were observed in bovine, human and sheep but not in horse erythrocytes. Proliferation following invasion was higher in human but lower in horse and sheep erythrocytes compared with bovine erythrocytes. Long-term cultures of B. divergens reached similar peak parasitaemias in human, sheep and bovine erythrocytes. Attempts to establish long-term cultures in horse erythrocytes failed. These results suggest that B. divergens is not host specific at the level of host cell attachment and invasion. Instead, parasite survival appears to be decided once the organism has gained access into the cell.

Animals↗
PubMed2002

Chymotrypsin and neuraminidase treatment inhibits host cell invasion by Babesia divergens (Phylum Apicomplexa).

The process of host cell invasion by Babesia divergens is poorly understood and improved knowledge of the mechanism involved could lead to development of measures effective in disease prevention. The investigate parasite ligands on the erythrocyte surface, B. divergens cultures in bovine erythrocytes were transferred into enzyme-treated bovine, human, ovine and equine erythrocytes. Parasite invasion of bovine erythrocytes was not affected by trypsin treatment while treatment with alpha-chymotrypsin led to a reduction in parasite growth of 20-40%. Treatment of bovine and non-bovine erythrocytes with neuraminidase decreased their susceptibility to invasion by up to 97% implicating sialic acid as an important erythrocyte ligand for babesia, but the addition of either bovine or human N-acetylneuraminyl-lactose to B. divergens cultures in bovine erythrocytes had no inhibitory effect.

Animals↗
PubMed2001

Extracellular polysaccharides of modified strains of Erwinia spp.

The structure of the extracellular polysaccharide (EPS) produced by Erwinia chrysanthemi strain A2148 has been determined using low pressure size-exclusion and anion-exchange chromatographies, high pH anion-exchange chromatography, glycosyl-linkage analysis, and 1D 1H NMR spectroscopy. The polysaccharide is structurally similar, if not identical, to the EPS produced by E. chrysanthemi strain A350. A streptomycin-resistant strain of E. chrysanthemi Ech6 (Ech6S(+)) has been generated and has an elevated production of EPS, as does a streptomycin-resistant strain (Ech9Sm6) of E. chrysanthemi Ech9. These modified E. chrysanthemi spp. have been ribotyped and found to be closely related to their parent strains.

Carbohydrate Sequence↗
PubMed2001

Predicting effects of toxic chemicals in the marine environment.

Ecological risks are typically characterized in risk assessment procedures by considering the ratio between exposure concentrations and critical effect concentrations. In OECD countries, critical effect concentrations are typically derived from laboratory-based ecotoxicity tests using well-defined protocols on a limited number of species. More and more countries in the tropics are adopting this approach in environmental assessment, protection, and management. In this article we consider a number of issues associated with such an approach, and in particular potential problems with extrapolating effects on individuals observed in laboratory-based ecotoxicological investigations to effects on ecosystems. It is hoped that by making explicit some of the assumptions made in the potential limitations of these tests, we can better target our limited resources to protect valuable and vulnerable systems.

Environmental Monitoring↗
PubMed2000

Extracellular polysaccharide of Erwinia chrysanthemi A350 and ribotyping of Erwinia chrysanthemi spp.

Erwinia chrysanthemi spp. are gram-negative bacterial phytopathogens causing soft rots in a number of plants. The structure of the extracellular polysaccharide (EPS) produced by the E. chrysanthemi strain A350, which is a lacZ- mutant of the wild type strain 3937, pathogenic to Saintpaulia, has been determined using a combination of chemical and physical techniques including methylation analysis, low-pressure gel-filtration and anion-exchange chromatography, high-pH anion-exchange chromatography, partial acid hydrolysis, mass spectrometry and 1- and 2D NMR spectroscopy. In contrast to the structures of the EPS reported for other strains of E. chrysanthemi, the EPS from strain A350 contains D-GalA, together with L-Rhap and D-Galp in a 1:4:1 ratio. Evidence is presented for the following hexasaccharide repeat unit: [structure: see text] All the Erwinia chrysanthemi spp. studied to date have been analyzed by ribotyping and collated into families, which are consistent with the related structures of their EPS.

Carbohydrate Conformation↗
PubMed2000

A strategic dosing scheme for the control of fasciolosis in cattle and sheep in Ireland.

This study examined the effectiveness of a strategic dosing scheme in lowering the incidence of fasciolosis on a mixed dry-stock farm and in maintaining the reduced incidence following a reduction in dosing intensity. Two neighbouring farms with a history of chronic fluke disease were selected, the strategic dosing scheme being implemented on one (the trial farm) while the other (the control farm) continued to treat according to its normal practice. The strategic dosing scheme was designed to suppress the faecal egg output of Fasciola hepatica at critical times of the year in order to limit infection of the intermediate host snail population and thus reduce the subsequent contamination of the pasture with metacercariae. On the trial farm cattle and sheep were treated three times per year for the first 2 years at approximately 8 week intervals, starting in March of each year. A fourth treatment was given when the cattle were housed and out-wintered sheep received an additional treatment in January. In Years 3 and 4 the dosing intensity was reduced. By the end of Year 2, data from faecal egg counts, tracer-sheep fluke burdens and snail infection levels indicated that the treatment strategy had succeeded in suppressing the fluke population and eliminating the occurrence of clinical fasciolosis. The decrease in dosing intensity in Years 3 and 4 maintained both stock and snail infections at low levels and there was no re-emergence of the disease.

Animals↗
PubMed2000

Tick bite and Lyme borreliosis risk at a recreational site in England.

The risk of tick bite and Lyme borreliosis in a forested area in England with public access was studied over a two-year period. Tick infestation levels were high with more than 1000 members of the public reporting for tick removal at a local clinic. Most of the attached ticks were nymphs (82%) and distinct differences in anatomical sites of attachment were observed in children and adults. Children sustained nymphal bites to the head, neck and axilla region much more frequently than adults (48 vs. 10%), whereas adults were bitten on the lower legs more frequently than children (46 vs. 9%). The vegetation was heavily infested with ticks and high numbers were particularly associated with areas used by deer. The average density of nymphs collected from the vegetation was 14.1 per 10 m2 (range 5.1-43.6). Infection rates of these nymphs determined by PCR and indirect IFA ranged from 5.2-17.0%, and the genospecies Borrelia valaisiana and B. garinii were detected, suggesting that birds may be important reservoir hosts in this area. It is estimated that, at the level of tick challenge observed here, at least 50 persons per year may be bitten by infected ticks at this site. However, no cases of Lyme borreliosis have been reported through the clinic follow-up procedure, and sera from 19 forest workers were negative for antibody to B. burgdorferi sensu lato. Despite the high challenge from tick bites, this particular recreational forest site poses a low risk of infection to the general public, and prophylactic antibiotic treatment or serological testing following a bite is not justified.

Adult↗
PubMed1999

Autohydrolysis of phytic acid.

The autohydrolysis of phytic acid at 120 degrees C resulted in the formation of most of the phosphate esters of myo-inositol in varying amounts depending upon the reaction time. Eighteen of the 39 chromatographically distinct myo-inositol mono-, bis-, tris-, tetrakis-, pentakis-, and hexakisphosphates have been characterized using two different HPLC systems. These myo-inositol phosphates were partially purified by preparative anion-exchange chromatography under acidic and alkaline elution conditions. The combination of these two methods provides a two-tiered chromatographic approach to the rapid and sensitive identification of inositol phosphates in complex mixtures. Identification of the products was confirmed by 1D and 2D (1)H NMR analysis. The analytical procedure was applied to the autohydrolysis of the mixture of inositol phosphates from corn steep water.

Chromatography, High Pressure Liquid↗
PubMed1999

Extracellular polysaccharide of Erwinia chrysanthemi CU643.

Erwinia chrysanthemi are gram-negative bacterial phytopathogens causing soft rots in a number of plants. The structure of the extracellular polysaccharide (EPS) produced by E. chrysanthemi strain CU643, pathogenic to Philodendron, has been determined using a combination of chemical and physical techniques including methylation analysis, high- and low-pressure gel-filtration and anion-exchange chromatography, high-pH anion-exchange chromatography, partial acid hydrolysis, mass spectrometry, and 1- and 2-D NMR spectroscopy. In contrast to the structures of the EPS reported for other strains of E. chrysanthemi, the EPS from strain CU643 is a linear polysaccharide containing L-Rhap, D-Galp, and D-GlcAp in the ratio 4:1:1. Evidence is presented for the following hexasaccharide repeat unit: -->3)-beta-D-Galp-(1-->2)-alpha-L-Rhap-(1-->4)-beta-D-GlcAp- (1-->2)-alpha-L- Rhap-(1-->2)-alpha-L-Rhap-(1-->2)-alpha-L-Rhap-(1-->(1 ).

Carbohydrate Sequence↗
PubMed1999

Borrelia burgdorferi sensu lato in Ixodes ricinus ticks and rodents in a recreational park in south-western Ireland.

Ixodes ricinus ticks infected with Borrelia burgdorferi sensu lato were numerous on the edges of paths and roads in a recreational park in south-western Ireland. The abundance of ticks at different sites was related to the presence of deer, but a negative relationship was shown between tick abundance and tick infection rates. This is thought to be due to the deposition of large numbers of uninfected ticks by deer, which are apparently not good reservoir hosts of B. burgdorferi s.l. Blood meal analysis only detected deer DNA in uninfected nymphs. Reservoir competent rodents, Apodemus sylvaticus and Clethrionomys glareolus, were abundant at all sites and a high proportion of captured specimens were infested with larval ticks. However, very few rodents were infected with B. burgdorferi s.l. and none of the unfed infected nymphs analysed for the identity of their larval blood meal had fed on rodents. The spirochaetes detected in I. ricinus in the study area may be poorly adapted to rodents or are not transmitted readily because of the absence of nymphal infestation. The majority of spirochaetes in these ticks were apparently acquired from non-rodent hosts, such as birds.

Animals↗
PubMed1999

Activity of atovaquone against Babesia microti in the Mongolian gerbil, Meriones unguiculatus.

The hydroxynaphthoquinone, atovaquone (Wellvone, Glaxo-Wellcome Ltd.) was found to have significant activity against Babesia microti, the main cause of human babesiosis in the U.S.A. This activity compares well with that of the most effective babesicide currently available for use in animals, imidocarb dipropionate, that unlike atovaquone is not licensed for use in humans. Treatment with well tolerated doses of atovaquone results in a rapid reduction in parasitemias and an early disappearance of parasites from blood smears. However, in common with all the other babesicides tested, atovaquone did not sterilize gerbils of infection, even at very high daily doses administered for up to 10 days. A combination of atovaquone and clindamycin was more effective than atovaquone alone in the treatment of both acute and chronic infections but failed to eliminate parasites completely.

Acute Disease↗
PubMed1998

Risk assessment for complex chemical exposure in aquatic systems: the problem of estimating interactive effects.

The traditional, but little used, way of assessing effects of the interaction between known chemicals is to use factorial experimental designs. Such designs allow one to test for less than additive (antagonistic) and greater than additive (synergistic) effects. Whilst synergism can be demonstrated in such experiments the concentrations at which synergistic effects occur are extremely high and are unlikely to occur in nature. Recently developed techniques allow one to measure directly the effects of combined stressors in the field. These biological effect techniques range from tests on individual organisms to tests on communities. At the biochemical level the tests can indicate that the organism has been exposed to certain groups of chemicals (for example cytochrome P-450 enzymes responding to PAHs or metallothioneins responding to heavy metals). At the community level of organisation there are highly sensitive statistical techniques that indicate clearly the combined effect of stressors. The effects of oil exploration and production on benthic communities in the North Sea can be linked to concentrations of chemicals. However, such relationships are correlative and do not necessarily indicate cause and effect. Experiments are needed to test the hypotheses generated concerning the interactive effects of chemicals on the benthic species. The statistical analyses do, however, show which species have been affected and their relative sensitivity to chemical and physical disturbances. Such species are preferable to the traditional "laboratory weeds" usually utilised. A strategy for risk assessment is needed that combines an experimental protocol for making predictions, from laboratory experiments, of likely effects to be found in the field. This should be combined with field monitoring that allows one to detect changes that were not predicted. At present most monitoring designs cannot adequately detect trends. This is due to concentration on Type-I statistical errors rather than properly considering Type-II errors. By concentrating on Type-II errors one can design monitoring programmes that are able to detect trends with a given degree of precision. There are also strong ethical grounds for a change to giving more emphasis to Type-II errors.

Animals↗
PubMed1998

Heterogeneity of glycans at each N-glycosylation site of horseradish peroxidase.

The tryptic glycopeptides of horseradish peroxidase isozyme c (HRPc) were studied by methylation linkage analysis, exoglycosidase degradation, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOFMS). Over 90% of the predicted tryptic peptides and glycopeptides of HRPc could be identified in the unfractionated digest. Four glycans, namely (Xyl)Man3(Fuc)GlcNAc2 (major species), (Xyl)Man2(Fuc)GlcNAc2, (Xyl)Man3GlcNAc2, and Man3(Fuc)GlcNAc2 (minor species), were observed at all of the N-glycosylation sites and account for greater than 95% of the carbohydrate. Other members of this glycan family, namely (Xyl)xManm(Fuc)f GlcNAc2 (x = 0 or 1, f = 0 or 1, m = 4, 5, 6, or 7), account for the rest of the glycans. Only traces of high mannose-type glycans were detected in HRPc. Two sites, namely those at Asn-57 and Asn-267, were found to be more heterogeneous than the sites at Asn-13, Asn-158, Asn-186, 198 (doubly glycosylated peptide), Asn-214, and Asn-255. Two of the glycopeptides were observed as part of disulfide-linked species. MALDITOFMS confirmed the N-glycosylation sites previously reported [K.G. Welinder, Eur. J. Biochem., 96 (1979) 483-502] and was used to determine the heterogeneity of the glycan pool at each site.

Amino Acid Sequence↗