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Biomedical subjects

D L Lee

Publications and source records attributed to D L Lee.

At least 55 records · Page 3Linked to original sources
PubMed1993

Acetylcholinesterase in infective-stage larvae of Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis resistant and susceptible to benzimidazole anthelmintics.

The amounts of acetylcholinesterase (AChE) in homogenates of infective-stage larvae of strains, from different countries, of Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis resistant or susceptible to benzimidazole (BZ) anthelmintics, were compared using a rapid colorimetric assay. The levels of AChE were much greater in those strains which were BZ-resistant than in BZ-susceptible populations. The larvae of susceptible and resistant H. contortus contained less AChE than did the larvae of the other two species. One-way analysis of variance showed that the observed differences between susceptible and resistant strains were significant in each of the species studied. It is suggested that the extra enzyme may act as an anthelmintic-detoxifying enzyme, may inactivate the active site of the anthelmintic or may prevent binding of the anthelmintic to its receptor.

Acetylcholinesterase↗
PubMed1993

Biochemical characterisation of Trichinella spiralis and T. pseudospiralis stichocyte antigens.

One and two dimensional Western blots of Trichinella spiralis and Trichinella pseudospiralis extracts were probed with a monoclonal antibody (7C2C5) which recognizes molecules found in the stichocytes. Three major antigens were revealed in excreted:secreted (E/S) preparations at approx Mr 45, 53 & 60 X 10(3). The Mr 45 X 10(3) molecule was the most abundant secreted antigen, on two dimensional analysis it resolved into many isoforms with a pI range 4.5-6.4 in T. spiralis and into four isoforms pI range 5.1-5.7 in T. pseudospiralis.

Animals↗
PubMed1992

Wastewater reuse in agriculture and the risk of intestinal nematode infection.

The increasing use of wastewater for crop irrigation in arid and semi-arid zones means that wastewater parasitology is becoming on important research area. The World Health Organization's (WHO) current guideline states that only treated wastewater containing no more than one human intestinal nematode egg per litre should be used for irrigation. This guideline was based on very limited epidemiological evidence. In this article Rachel Ayres, Rebecca Stott, Duncan Mara and Donald Lee describe their recent work in Yorkshire and northeast Brazil and suggest that their results may be the first evidence for relaxing the WHO guideline for restricted irrigation to no more than ten intestinal nematode eggs per litre.

Journal Article↗
PubMed1991

The presence of peptides related to the adipokinetic hormone/red pigment-concentrating hormone family in the nematode, Panagrellus redivivus.

Immunocytochemistry using polyclonal antisera raised to fragments or derivatives of locust adipokinetic hormone (AKH) I and IIs (Schooneveld et al., 1983, 1985, 1986) selectively stained cells in the nervous system of the free-living nematode, Panagrellus redivivus. Antiserum 528 (raised to the C-terminus of AKH IIs) stained the dorsal cephalic papillary cell bodies and the anterior nerve ring. Fibres in the lateral cords were stained with antiserum 241 that recognises the C-terminus of AKH I. Substances reacting to antisera 433 (raised to the N-terminal sequence of AKH I and IIs) 528 and 241 were present in the preanal ganglion and associated ventral nerve fibres. In males, all three antisera stained fibres leading to the base of the spicules. A peptide fraction from whole P. redivivus evoked an adipokinetic response in the locust, Schistocera gregaria which was dose dependent and was abolished by treatment with endopeptidase 24:11 but not by boiling or by incubation with leucine aminopeptidase. The adipokinetic activity was reduced by over 70% on incubation of the peptide fraction with pyroglutamyl aminopeptidase. The same fraction induced hyperglycaemia when injected into the cockroach, Periplaneta americana. These results are consistent with the existence in P. redivivus of peptides that are structurally related to the arthropod adipokinetic hormone/red pigment-concentrating hormone (AKH/RPCH) family.

Amino Acid Sequence↗
PubMed1991

Trichinella spiralis: antigenic epitopes from the stichocytes detected in the hypertrophic nuclei and cytoplasm of the parasitized muscle fibre (nurse cell) of the host.

Monoclonal antibodies raised against antigens present in the excretions/secretions (E/S) of larval Trichinella spiralis, polyclonal antibodies raised against E/S and antisera from rabbits and pigs infected with T. spiralis were used in conjunction with immunocytochemical techniques to detect antigens in sections of muscle from mice that had been infected with T. spiralis for 15 or 30 days. The antibodies recognized epitopes in the stichocytes, on the surface of the cuticle, in the lumen of the oesophagus and in the lumen of the intestine of encysted larvae. Monoclonal antibodies 7C2C5 and 1H7 and the polyclonal antibodies recognized epitopes in the cavity occupied by the larva, in the cytoplasm of the nurse cell, and in the hypertrophic nuclei of the nurse cell, but did not recognize material in the smaller nuclei of the nurse cell, in the cyst wall or in the surrounding muscle. Monoclonals 3B2E6 and 1D11G8B2, which recognized epitopes in the stichocytes and on the surface of the cuticle of the larvae, gave negative results with the cytoplasm and nuclei of the nurse cell. A polyclonal antibody raised against Trichuris suis recognized epitopes in the muscle and hypodermis of encysted T. spiralis but gave negative results with material in the nurse cell and nurse cell nuclei. The possibility that the antigen detected in the cytoplasm and nuclei of the nurse cell is produced by the stichocytes of the nematode and that it is controlling genes of the altered muscle fibre, either directly or indirectly, is discussed.

Animals↗
PubMed1990

A copia-like element in Pisum demonstrates the uses of dispersed repeated sequences in genetic analysis.

A DNA sequence between two legumin genes in Pisum is a member of the copia-like class of retrotransposons and represents one member of a polymorphic and heterogeneous dispersed repeated sequence family in Pisum. This sequence can be exploited in genetic studies either by RFLP analysis where several markers can be scored together, or the segregation of individual elements can be followed after PCR amplification of specific members.

Base Sequence↗
PubMed1990

A larval paralysis assay for the detection of thiabendazole resistance in trichostrongyles.

Infective-stage larvae of trichostrongyle nematodes, either resistant or susceptible to thiabendazole (TBZ), were incubated in eserine, an acetylcholinesterase inhibitor. Paralysis occurred in larvae treated with eserine but significant differences were observed in the percentage of larva immobilized between TBZ-resistant and TBZ-susceptible strains of the nematodes. These differences are probably related to the presence of higher levels of acetylcholinesterase in the TBZ-resistant strains than in the susceptible strains. This could be used as a rapid and inexpensive method of detecting resistance to TBZ in trichostrongyles.

Animals↗
PubMed1990

Two- and three-dimensional locomotion of the nematode Nippostrongylus brasiliensis.

Locomotion of adult Nippostrongylus brasiliensis has been studied in saline, in 0.6% agar, in sodium alginate of different viscosities and amongst sand grains in these media. In saline the nematode formed two-dimensional waves but there was little forward progression. Amongst sand grains in saline the nematode moved forwards by thrusting against sand grains, but thigmokinetic behaviour later resulted in quiescence. In 0.6% agar and in alginates of weak viscosity the nematode produced two-dimensional waves and sometimes a three-dimensional helical wave which resulted in forward movement. The formation of three-dimensional waves and the distance travelled increased with increasing viscosity up to 4% sodium alginate and also amongst sand gains in these media. In 8% sodium alginate the nematode became coiled like a spring but remained almost stationary. The three-dimensional wave is formed with torsion and obtains thrust from the viscous medium. In the intestine of the host thrust will be obtained from the mucus and villi of the intestinal mucosa. The ability of this nematode to move by two- and three-dimensional undulatory propulsion is probably related to its complex ridged cuticle. Attention is drawn to the role that increased viscosity of mucus may play in entrapping nematodes during their immune rejection.

Animals↗
PubMed1989

Post-mortem changes in the abomasal mucosae of sheep infected with Haemonchus contortus compared with those in uninfected sheep.

Progressive post-mortem changes were observed, with fibre optic endoscopy, in the abomasum of uninfected sheep and sheep infected with H. contortus. Loss of epithelial cells was observed 14 to 16 min after death in the uninfected sheep. The loss exposed the underlying lamina propria. This loss became more widespread as the time after death increased. Epithelial cell loss was observed earlier (as early as 6 min after death) in the infected sheep. Three of the infected sheep also displayed dispersal of alkaline phosphatase activity 30 min after death. The present work shows that post-mortem changes can occur quite quickly in animals, especially infected animals, and emphasizes the importance of rapid removal of tissue from animals after death.

Abomasum↗
PubMed1989

Colorimetric assay for the detection of benzimidazole resistance in trichostrongyles.

A modified version of the aphid tile-test, which is used to detect insecticide resistance in single adult aphids that are resistant or susceptible to organophosphate or carbamate insecticides, was used to compare the levels of non-specific esterases in strains of the trichostrongyle nematodes Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis which were known to be resistant or susceptible to benzimidazole (BZ) anthelmintics. This colorimetric assay has shown that there is significantly more non-specific esterase in the infective-stage larvae of BZ-resistant strains than in susceptible strains and this may prove to be of use in the detection of resistance to benzimidazole anthelmintics.

Animals↗
PubMed1988

Detection of benzimidazole resistance in trichostrongylid nematodes.

Benzimidazole anthelmintics (Box 1) have an important role in the control of economically-important nematode parasites of sheep and other ruminants. But effective control and management strategies using such compounds require a clear appraisal of the resistance or susceptibility of the target worms. Currently available tests for benzimidozole resistance in trichostrongylid nematodes tend to be costly, time-consuming and technically-demanding - as a result, attention is now turning to biochemical assays.

Journal Article↗
PubMed1988

Immunocytochemical demonstration of a neuropeptide in Ascaris suum (Nematoda) using an antiserum to FMRFamide.

A FMRFamide-like peptide has been detected in the nematode Ascaris suum, using the peroxidase-anti-peroxidase (PAP) immunocytochemical technique. Positive reactions were obtained in both the central nervous system and the peripheral nervous system of the worm, the strongest reactions being in the anterior nerve ring, the cephalic papillary ganglia, the lateral ganglia and the dorso-rectal ganglion. Immunoreactivity was observed along the length of the main nerve cords of the worm and, to a lesser extent, in the pharyngeal nerve cords. The possible role of this neuropeptide in the physiology of the nematode is discussed.

Animals↗
PubMed1988

Primary and secondary infection of the domestic chicken with Trichostrongylus tenuis (Nematoda), a parasite of red grouse, with observations on the effect on the caecal mucosa.

The course of primary and secondary infections with Trichostrongylus tenuis in the domestic chicken was investigated. Primary infections were established after the administration of single and trickle doses of infective-stage larvae. The worm burden in the caeca was highest after a single dose of 500 infective-stage larvae; this gave a mean of 87 nematodes per bird on days 89 of infection, 20 nematodes on day 14 and 0 on day 28 of infection. Following trickle doses of 60, 100, 200, 300, 400 or 500 infective-stage larvae, there was a rise and then a fall in nematode egg output in all groups. In chickens given a primary dose of 500 infective-stage larvae followed 30 days later by a single secondary dose of 500 infective-stage larvae, the mean worm burden during the secondary infection rose to 57 nematodes on day 9 of infection and then fell rapidly to 18 nematodes on day 15 and to 2 on day 30. Scanning electron microscopy showed changes in the caeca of infected birds, with the caecal surface being covered in a layer of mucus from 12 days after infection. Ball of blood-stained mucus containing nematodes were observed in the caecal droppings from day 9 of infection onwards. It is concluded that chickens rapidly expel an established infection of T. tenuis, unlike the normal host, the red grouse.

Animals↗
PubMed1988

Impaired cardiac muscarinic receptor function in dogs with heart failure.

Prior physiological studies have suggested that parasympathetic control is altered in heart failure. The goal of our studies was to investigate the influence of heart failure on the muscarinic receptor, and its coupling to adenylate cyclase. Ligand binding studies using [3H]quinuclidinyl benzilate and enriched left ventricular (LV) sarcolemma, demonstrated that muscarinic receptor density in heart failure declined 36% from a control of 5.6 +/- 0.6 pmol/mg, with no change in antagonist affinity. However, agonist competition studies with both carbachol and oxotremorine showed that it was a loss of high affinity agonist binding sites in the sarcolemma from failing LV that accounted for this difference. The functional efficacy of the muscarinic receptor was also examined. When 1 microM methacholine was added to 0.1 mM GTP and 0.1 mM isoproterenol, adenylate cyclase stimulated activity was inhibited by 15% in normal LV but only 5% in LV sarcolemma from animals with heart failure even when the reduced adenylate cyclase in these heart failure animals was taken into account. Even at 100-fold greater concentrations of methacholine, significantly less inhibition of adenylate cyclase activity was observed in LV failure as compared with normal LV sarcolemma. Levels of the GTP-inhibitory protein known to couple the muscarinic receptor to adenylate cyclase, as measured with pertussis toxin labeling, were not depressed in LV failure. Thus, the inhibitory pathway regulating LV adenylate cyclase activity is defective in heart failure. The decrease in muscarinic receptor density, and in particular the specific loss of the high affinity agonist binding component of this receptor population, appears to be the major factor underlying this abnormality.

Adenylyl Cyclases↗
PubMed1988

Hypergastrinaemia of sheep infected with Haemonchus contortus.

Radioimmunoassay of plasma gastrin has shown that a hypergastrinaemia occurs in yearling sheep, after infection with 10(6) infective third stage larvae of Haemonchus contortus. Plasma gastrin concentrations began to rise two to four days after infection, that is, at about the same time as, or slightly before, the abomasal pH was observed to increase.

Abomasum↗
PubMed1987

Physiological and microbiological changes in the abomasum of sheep infected with large doses of Haemonchus contortus.

The pH, PO2 and the number of viable bacteria per ml of abomasal fluid were recorded before and after administering large doses of infective larvae of H. contortus to sheep. Highly significant increases were observed in the pH, in numbers of viable bacteria and in the proportion of oxygen-sensitive bacteria in the abomasal fluid during the first 10 to 14 days of infection. At the same time a decrease in the abomasal fluid PO2 was recorded. Subsequently a slow recovery of pH, PO2 and bacterial numbers to pre-infection values ensued. The microbiological and physiological changes occurring in the abomasum of sheep after infection with H. contortus are discussed with reference to these results.

Abomasum↗