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Biomedical subjects

D E Thompson

Publications and source records attributed to D E Thompson.

At least 19 recordsLinked to original sources
PubMed1992

Studies on the ribosomal RNA operons of Listeria monocytogenes.

A 23S rRNA gene of Listeria monocytogenes was cloned into pUC19 on a 6.2-kb Pst I fragment. Hybridisation studies demonstrated the presence of the 5S and partial 16S rRNA genes within the clone. The nucleotide sequence of the region encoding the 23S rRNA was found to be highly homologous with those of other low G + C Gram-positive bacteria. The 16S-23S intergenic spacer region was amplified using PCR technology and revealed two product sizes, the larger of which contained tRNA(Ala) and tRNA(Ileu) genes. Further tRNA genes were found downstream of the 5S rRNA gene.

Base Sequence↗
PubMed1992

Sequence of the gene encoding type F neurotoxin of Clostridium botulinum.

Primers designed to conserved regions of botulinum and tetanus clostridial toxins were used to amplify DNA fragments from non-proteolytic Clostridium botulinum type F (202F) DNA using polymerase chain reaction technology. The fragments were cloned and the complete nucleotide sequence of the gene encoding type F toxin determined. Analysis of the nucleotide sequence demonstrated the presence of an open frame encoding a protein of 1274 amino acids, similar to other botulinum neurotoxins. Upstream of the toxin gene is the end of an open reading frame which encodes the C-terminus of a protein with homology to non-toxic-non-hemagglutinin component of type C progenitor toxin.

Amino Acid Sequence↗
PubMed1992

PCR cloning and nucleotide sequence determination of the 18S rRNA genes and internal transcribed spacer 1 of the protozoan parasites Cryptosporidium parvum and Cryptosporidium muris.

The genes encoding 18S rRNA and internal transcribed spacer 1 (1TS1) of Cryptosporidium parvum and Cryptosporidium muris were amplified from oocysts by PCR utilizing primers complementary to conserved regions of the 5' end of 18S and 5.8S rRNA. PCR products were cloned and the complete nucleotide sequences of two clones of each Cryptosporidium species were determined. The 18S rRNA genes of C. parvum and C. muris showed more than 99% sequence identity.

Animals↗
PubMed1992

Analysis of operons encoding 23S rRNA of Clostridium botulinum type A.

Southern hybridization analysis of Clostridium botulinum type A chromosomal DNA indicated the presence of six copies of the 23S rRNA gene. Fragments of DNA encoding 23S rRNA were amplified by polymerase chain reaction and cloned in Escherichia coli. Three clones examined by restriction enzyme and sequence analysis were found to be derived from different operons. Sequence determination of the entire insert of two clones revealed nine nucleotide changes in the genes coding for 23S rRNA (99.7% sequence identity) between operons encoded on the same chromosome, showing microheterogeneity in the rRNA operons of this organism.

Base Sequence↗
PubMed1992

In vitro cytotoxicity of melanized poly(2-hydroxyethyl methacrylate) hydrogels, a novel class of ocular biomaterials.

Due to their ability to absorb ultraviolet and visible radiation, we have proposed the melanized poly(2-hydroxyethyl methacrylate) hydrogels as biomaterials suitable for the manufacture of soft artificial intraocular lenses. Their biocompatibility has not been evaluated so far. In this study, poly(2-hydroxyethyl methacrylate) containing various amounts of adrenochrome-melanin were synthesized and the cytotoxicity of their aqueous extracts was assessed by using four in vitro testing techniques (trypan blue dye exclusion, inhibition of DNA synthesis, lactate dehydrogenase release, and inhibition of cell growth). Assays were based on incubation with human choroidal fibroblasts. By the first three techniques, no cytotoxicity was found in the extracts. The inhibition of cell growth test showed a slight cellular protein loss, however only in the extracts of polymers with high melanin content. The results suggest that the release of potentially toxic agents from melanized hydrogels into an aqueous medium is not significant. However, when an assay in collagen gel was carried out in the presence of specimens of melanized hydrogels, a toxic reaction was clearly revealed. This can be caused by a delayed release of toxic molecules from melanin, or by some other mechanism. The use of melanin-containing polymers as implant materials becomes questionable and further research is necessary.

Biocompatible Materials↗
PubMed1991

Cytotoxic effects of residual chemicals from polymeric biomaterials for artificial soft intraocular lenses.

Development of improved hydrogels for soft intraocular lenses, based on 2-hydroxyethyl methacrylate monomer, requires the use of various other monomers and polymerization additives which have potential ocular toxicity. Three monomers, 2-hydroxyethyl methacrylate, methyl methacrylate, and 2-ethoxyethyl methacrylate, as well as two common inhibitors, hydroquinone and 4-methoxyphenol, were subjected to in vitro cytotoxicity assays as aqueous solutions at different concentrations. A new polymerization initiator, 2,2'-azo-bis-(2,4-dimethyl valeronitrile), was thermally decomposed in water at different concentrations and the products were also assayed for cytotoxicity. Assays were based on incubation with human choroidal fibroblasts. Cell death was evaluated by trypan blue dye exclusion, DNA synthesis inhibition, and lactate dehydrogenase tests. While methyl methacrylate and 2-ethoxyethyl methacrylate were found nontoxic, the other chemicals displayed high cytotoxicity. However, when extracts of synthesized poly(2-hydroxyethyl methacrylate) specimens, differentially treated after polymerization, were subjected to the same assays it was found that toxicity from residual 2-hydroxyethyl methacrylate monomer was lost during steam sterilization and storage in water because of the removal of the monomer through aqueous washing. The lack of toxicity in these specimens suggests that residual contents of inhibitor and initiator are too low to cause toxic effects on choroidal fibroblasts. It is concluded that hydrogels have low cytotoxic effects in vitro.

Anisoles↗
PubMed1990

The complete amino acid sequence of the Clostridium botulinum type A neurotoxin, deduced by nucleotide sequence analysis of the encoding gene.

A 26-mer oligonucleotide probe was synthesized (based on the determined amino acid sequence of the N-terminus of the Clostridium botulinum type A neurotoxin, BoNT/A) and used in Southern blot analysis to construct a restriction map of the region of the clostridial genome encompassing BoNT/A. The detailed information obtained enabled the cloning of the structural gene as three distinct fragments, none of which were capable of directing the expression of a toxic molecule. The central portion was cloned as a 2-kb PvuII-TaqI fragment and the remaining regions of the light chain and heavy chain as a 2.4-kb ScaI-TaqI fragment and a 3.4-kb HpaI-PvuII fragment, respectively. The nucleotide sequence of all three fragments was determined and an open reading frame identified, composed of 1296 codons corresponding to a polypeptide of 149 502 Da. The deduced amino acid sequence exhibited 33% similarity to tetanus toxin, with the most highly conserved regions occurring between the N-termini of the respective heavy chains. Conservation of Cys residues flanking the position at which the toxins are cleaved to yield the heavy chain and light chain allowed the tentative identification of those residues which probably form the disulphide bridges linking the two toxin subfragments.

Amino Acid Sequence↗
PubMed1990

Physical characterisation of the replication region of the Streptococcus faecalis plasmid pAM beta 1.

The complete nucleotide (nt) sequence of a 5.1-kb EcoRI DNA restriction fragment carrying the replication region of the Streptococcus faecalis plasmid pAM beta 1 has been determined. Of the seven major open reading frames (ORF A-G) identified within this fragment, two (C and E) were shown to be encoding by in vitro transcription/translation assays. Evidence was obtained that synthesis of the polypeptide (Mr 57,380) encoded by the largest ORF (E) was essential for replication. Deletion analysis indicated that the minimum unit of DNA required for replication resided on a 2.59-kb AccI-HpaI subfragment. ORF C resided outside of this fragment and encompassed an extensive region of directly repeated nt sequence. The encoded polypeptide (Mr 30,471) was therefore composed of large tracts of reiterated amino acid sequence (11 x VDP and 35 x TEP tripeptides) which probably caused the observed anomalous electrophoretic mobility of the synthesised protein (equivalent to 61 kDa). Deletion of a 416-bp segment of DNA between unique KpnI and StyI sites caused an increase in copy number, which correlated with the in vitro production of higher levels of ORF E polypeptide. Although homology was detected between the sequenced DNA, and the replicon of a closely related streptococcal plasmid (pSM19035), none was evident to any other characterised Gram+ plasmid.

Amino Acid Sequence↗
PubMed1990

Planning tendon paths using an interactive graphic workstation.

This study provides a technique to be used for planning tendon paths in thumb reconstruction surgery. All mathematical modeling computations are performed on a VAX 11/750 host computer and the graphic manipulation is carried out by the Evans & Sutherland PS390 color display system. The results of the simulation are stored in a log file, including the rotation angles of the joints and the location of the pulley and the insertion points as a record of the tendon transfer design for a specific hand. The methods are based on the modeling of two separate types of tendon paths that consist of straight line segments and curved segments that follow bone contours. The method further assumes that the path of the tendon will always evolve to a planar curve. By integrating this technique with an existing kinematic model of the hand derived from CT-scans, a clinically relevant method has been developed.

Computer Graphics↗
PubMed1989

A kinematic model of the flexor tendons of the hand.

The multi-joint model is a kinematic simulation of the long flexor tendons of the fingers. The tendons modeled are the flexor pollicis longus, the flexor digitorum profundus, and the flexor digitorum superficialis. The simulated tendons are displayed on an Evans and Sutherland PS330 color graphics terminal attached to a display of articulated bones of the hand. As a user changes the position of the joints of the simulated hand, the simulation displays the new tendon path and the excursion of the tendon for the new position of the hand. The multi-joint model is one component of a comprehensive model for use in a hand biomechanics computer workstation.

Computer Graphics↗
PubMed1988

Introduction of genes for leucine biosynthesis from Clostridium pasteurianum into C. acetobutylicum by cointegrate conjugal transfer.

A Clostridium pasteurianum gene bank was constructed in Escherichia coli, using plasmid pAT153, and several chromosomal fragments found which complemented both leuB and leuC mutations in auxotrophic E. coli K12 strains. No fragments capable of complementing leuA or leuD mutations were identified. Conjugal transfer of the leuB/leuC genes from Bacillus subtilis into two different Leu- C. acetobutylicum auxotrophic strains was elicited by their incorporation into a large plasmid cointegrate composed of the conjugal plasmid pAM beta 1 and a specially constructed gram-positive, replication-deficient plasmid, pMTL21 EC. Inheritance of the cointegrate plasmid restored one of the auxotrophic C. acetobutylicum strains to prototrophy. The second strain remained Leu-.

3-Isopropylmalate Dehydrogenase↗
PubMed1988

Effect of age and initial infection intensity on the rate of reinfection with Trichuris trichiura after treatment.

The study examines the rate of re-acquisition of Trichuris trichiura infection after treatment in two populations, one of mixed age and the other of children with known pre-intervention infection intensity. A population living in a Caribbean village was treated with mebendazole and the rate of reacquisition of infection of four age classes (2-4, 5-10, 11-30, 30+ years) monitored over a 20-month period. The reinfection rate was higher in the child age-classes indicating either that children are more exposed to infection or that adults slowly develop a partially effective acquired immunity. A cohort of children (mean age 4.5 years) was separated into 3 intensity categories on the basis of expelled worm burdens and their rate of reacquisition of infection monitored over a 12-month period. The rate of reinfection was directly and positively associated with initial infection status. This may indicate that children with low intensity infections are consistently less exposed to infection or that they have effective immune responses. The latter conclusion, however, would imply that they had acquired this immunity early in life, and so appears to contradict the suggestion that resistance is only slowly acquired by adults. Reconciling these two conclusions may require more sophisticated immunological models than have been suggested previously for geohelminthiases.

Adolescent↗
PubMed1987

Age-related prevalence and intensity of Trichuris trichiura infection in a St. Lucian community.

Age-related changes in the average worm burden and the prevalence of Trichuris trichiura infection, in a village community in St. Lucia, were examined by field studies based on worm expulsion techniques. Horizontal age-intensity profiles were convex in form with peak parasite loads occurring in the 2 to 15-year-old children. Prevalence is shown to be a poor indicator of changes in average worm load with age. Faecal egg counts (epg and epd) provide a qualitative measure of worm burdens since fecundity is shown to be approximately independent of worm load. The parasites were highly aggregated within the study community, with most people harbouring low burdens while a few individuals harboured very heavy burdens. Of the total parasite populations in the study sample, 84% were harboured by the 2 to 15-year-old children. Of those individuals harbouring 100 worms or more, 87% were in the 2 to 10-year-old age range. Crude estimates of population parameters (basic reproductive rate, 4-5; rate of reinfection, 90 year-1) suggest that the rate of reinfection is higher than for other helminth parasites of man. The control of morbidity and parasite transmission is discussed in the context of targeting drug treatment at the child segment of the study population.

Adolescent↗
PubMed1987

Epidemiology and population dynamics of Ascaris lumbricoides and Trichuris trichiura infection in the same community.

The gastrointestinal helminth infection status of an age-stratified sample from a single Caribbean community was assessed using anthelmintic expulsion techniques. The same sample was re-assessed in a similar manner after a 17 month period of re-infection. The age-prevalence profile of Ascaris lumbricoides was convex while that of Trichuris trichiura was asymptotic. The age-intensity profiles of both species were convex. These differing patterns are attributed to differences in the absolute worm burdens of the 2 species. The frequency distributions of infection intensity were similar for both species, and largely independent of host age. The basic reproductive rate of A. lumbricoides (Ro = 1-1.8) was similar to that recorded elsewhere and much lower than that of T. trichiura (Ro = 4-6), implying that the latter is intrinsically more resistant to control. Individual hosts were predisposed to high (or low) intensity infection with either species, although predisposition to both species simultaneously was not conclusively demonstrated. Further studies are required to determine the cause of these observations.

Adolescent↗
PubMed1987

Predisposition to Trichuris trichiura infection in humans.

The study examines the distribution of Trichuris trichiura infection in a village community in St Lucia, West Indies. The infection intensity of the same age-stratified population was assessed (by drug expelled worm burden and faecal egg count) at the initiation of the study, and after 17 months of reinfection following treatment. The frequency distribution of worm numbers per person was similar at both periods of sampling. There was a significant correlation between the initial infection intensity of an individual, and the intensity acquired by the same individual following the 17 month period of reinfection. This relationship was observed in a broad range of host age classes. The study provides firm evidence that individuals are predisposed to heavy (or light) T. trichiura infection.

Adolescent↗
PubMed1987

Age-relationships of Toxocara canis seropositivity and geohelminth infection prevalence in two communities in St. Lucia, West Indies.

Sera were examined from an age-stratified sample of two Caribbean communities using the Toxocara-ELISA with larval ES antigen. Seropositivity was markedly age dependent, attaining maximal values (40 and 60%) in 5-15 year olds and declining in adults. The rate of acquisition of infection with Toxocara canis and the age-prevalence profile are similar to those of Ascaris lumbricoides and Trichuris trichiura. It is suggested that toxocariasis is likely to be prevalent in tropical areas with endemic geohelminthiasis.

Adolescent↗