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Biomedical subjects

J E Thomas

Publications and source records attributed to J E Thomas.

At least 19 recordsLinked to original sources
PubMed1992

Isolation of Helicobacter pylori from human faeces.

Helicobacter pylori is arguably the commonest chronic infection in man. However, its route of transmission is unknown. We have isolated viable H pylori from the faeces of an infected individual from The Gambia. The organism was cultured on selective media after concentration of faecal bacteria by centrifugation in a buffer equilibrated with a microaerophilic gas mixture. Growth characteristics, microscopic appearances, and enzyme activities were the same as those of a typical gastric isolate of H pylori. Protein preparations derived from the new isolate and the typical strain were antigenically similar, and had very similar electrophoretic profiles (including two major protein bands of 62 and 26 kDa, corresponding to the urease enzyme subunits). With the same technique, organisms with the colony morphology, growth requirements, enzyme activities, and microscopic appearances of H pylori were isolated from the faeces of 9 of 23 randomly selected children aged 3-27 months from a Gambian village with a high prevalence of H pylori infection in early life. Faecal-oral transmission is probably important in the spread of infection in such communities.

Adult↗
PubMed1992

The nucleotide sequence of the infectious cloned DNA component of tobacco yellow dwarf virus reveals features of geminiviruses infecting monocotyledonous plants.

An infectious clone of the Australian geminivirus tobacco yellow dwarf virus (TobYDV) was constructed from virus-specific double-stranded DNA isolated from infected tobacco and used to demonstrate a single-component genome. The nucleotide sequence of TobYDV DNA comprises 2580 nucleotides. TobYDV DNA has three coding regions, two in the virion sense and one in the complementary sense, homologous to those identified for other geminiviruses, particularly those infecting monocotyledonous (monocot) plants. The complementary sense coding region is comprised of two overlapping reading frames, with an intron of 86 nucleotides. Efficient splicing of the mRNA for this coding region was observed in the infected dicotyledonous (dicot) hosts bean and tobacco despite the intron having an A + U content (57%) more typical of geminiviruses of monocot plants. TobYDV encapsidates a small oligonucleotide able to prime synthesis of the complementary DNA strand in vitro. The TobYDV genome organization, low A + U intron, and encapsidated oligonucleotide primer resemble those of the monocot-infecting geminiviruses. These results strongly suggest that TobYDV is a monocot geminivirus which has become adapted to dicot hosts.

Base Sequence↗
PubMed1992

Oncogenes in X-ray-transformed C3H 10T1/2 mouse cells and in X-ray-induced mouse fibrosarcoma (RIF-1) cells.

In order to better understand the molecular basis of X-ray induced carcinogenesis we have investigated RNA levels of oncogenes in an X-ray transformed C3H 10T1/2 fibroblast line (XTD) and RIF-1 cells isolated from an X-ray-induced fibrosarcoma in a C3H mouse. Steady-state levels of K-ras, H-ras, N-ras, abl, sis, src, and fos were unchanged in the X-ray-transformed cells compared with non-transformed C3H 10T1/2 cells. However, myc and raf mRNA levels were increased dramatically in the transformed cells. Data further suggests a possible alteration in processing of raf RNA in the XTD cells. Southern blot analysis of secondary transfectants induced with XTD DNA indicated that the oncogenic phenotype did not segregate with the myc or raf loci; nor with nine other oncogenes analysed.

Animals↗
PubMed1992

Assay of urease-inhibiting activity in serum from children infected with Helicobacter pylori.

In order to provide a basis for obtaining further information concerning the host response to Helicobacter pylori urease, four assay methods for detecting urease-inhibiting activity in serum were examined. A quantitative assay, established in a COBAS BIO centrifugal fast analyzer and based on detection of the consumption of NADH by glutamate dehydrogenase stimulated by ammonia production, was considered most suitable for large-scale serological work. Serum samples from 63 children (aged 5 to 16 years), 28 of whom had seropositive H. pylori gastritis, were assayed. One of the serum samples in this latter group showed significant inhibitory activity. This serum sample was one of 13 in the seropositive group known to bind to urease antigen. It showed no inhibitory activity against Bacillus pasteurii or jack bean urease. Protein A binding and heat treatment indicated that the inhibitory activity was immunoglobulin G mediated. The patient from whom this sample was collected showed no distinctive features in his illness. The COBAS BIO analyzer-based urease inhibition assay provides a new tool for studying one aspect of the host response to H. pylori infection.

Child↗
PubMed1992

Amino acid pools in developing Chlamydomonas reinhardtii: vegetative cells, gametes, and mature zygotes.

Free amino acid pools were examined for cultures of vegetative cells, gametes, and mature zygotes of the unicellular green alga Chlamydomonas reinhardtii (Dangeard). The total pool of amino acids found in premature gametes of strains 137c+ (10.0 pmol-micrograms protein-1) and 137c- (10.8 pmol.micrograms protein-1) decreased to levels about half that seen in vegetative 137c- cells (19.8 pmol.micrograms protein-1). Following light activation, amino acid pools in these gametes increased to 18.7 pmol.micrograms protein-1 in 137c+ cells and 20.0 pmol.micrograms protein-1 in 137c- cells. With the exception of cystine, individual amino acid pools in these cells had increased once more to levels similar to those seen in vegetative cells grown in liquid medium. Levels of cystine remained one to two orders of magnitude lower than that seen in vegetative cells. Mature 137c+ and 137c- gametes mixed in solutions of either 2 mM cystine or 2 mM cysteine (half-cystine) suffered a 52-64% reduction, respectively, in the number of vis-à-vis conjugative pairs formed. This suggests that pools of endogenous cystine may play a role in the onset of mating. In zygotes levels of all amino acid pools, except histidine, were depressed; levels of cystine, valine, and phenylalanine were nondetectable in these cells.

Amino Acids↗
PubMed1992

A physician's guide to early detection of oral cancer.

Oral cancer is a disease of the elderly that is more prevalent in patients who drink alcohol, smoke, or are chronically exposed to the sun. An oral examination is recommended annually for all elderly patients and more often for patients with these risk factors. The key to diagnosis and long-term patient survival is recognition of early lesions, which are often asymptomatic. Cancerous lesions occur more frequently at specific oral sites and may be difficult to differentiate from lesions caused by such benign conditions as denture irritation and herpetic ulcers.

Aged↗
PubMed1991

Phosphorylation of c-Src on tyrosine 527 by another protein tyrosine kinase.

The protein tyrosine kinase activity of the cellular Src protein is negatively regulated by phosphorylation at tyrosine residue 527 (Tyr527). It has not been established whether this regulatory modification of Src is mediated by autophosphorylation or by another cellular protein kinase. The phosphorylation of a modified form of c-Src that lacks kinase activity was examined in mouse cells that do not express endogenous Src (because of the targeted disruption of both src alleles). Phosphorylation of the inactive form of Src on Tyr527 occurred to a similar extent in cells lacking endogenous Src as it did in cells expressing Src. Therefore, Tyr527 phosphorylation, and thus negative control of Src kinase activity, is mediated by another cellular protein tyrosine kinase.

Amino Acid Sequence↗
PubMed1991

Altered oncogenes in UV-transformed C3H 10T1/2 mouse cells: identification of mutated H-ras allele(s).

Ultraviolet (UV) light will transform mammalian cells in culture to a phenotype which is characteristic of in vivo neoplasia. The UV-transformed C3H 10T1/2 mouse cell lines, TU-2 and TU-3, were analysed to determine the molecular mechanisms which may account for their phenotype, and to determine the types of mutations induced by UV light. DNA-transfection assays indicated that the transformed phenotype of TU-2 could not be transferred to non-transformed recipient cells. Therefore, studies were initiated to determine the mutagenic effects of UV light with respect to cellular oncogenes. Northern blot analysis indicated that five of the oncogenes analysed (erb-A, erb-B, mos, myb, and N-ras) were not expressed at detectable levels. The steady-state mRNA levels of fos, K-ras, abl, sis, and src oncogenes were similar in the C3H 10T1/2 and TU-2 cells. The mRNA levels of three oncogenes, raf, myc and H-ras, were 1.5-2.0-fold greater in the TU-2 cells compared to C3H 10T1/2. Southern blot analysis of HpaII restriction digested TU-2 DNA indicated that the H-ras oncogene has undergone methylation changes. More extensive analyses of the H-ras locus in TU-2 demonstrated a deletion of the 3' end of the gene, that may involve two separate mutated alleles. This type of damage is consistent with the lesions associated with sister chromatid exchange. While the H-ras locus in the other UV-transformed line, TU-3, showed methylation changes, there were no large genetic mutations detected by Southern blot analysis. These results suggest that UV-irradiation in vitro induces endogenous DNA damage that includes methylation changes and large genomic alterations. Further analysis will be necessary to determine the extent to which each may be involved in cell transformation.

Alleles↗
PubMed1991

Purification, characterization and serological detection of virus-like particles associated with banana bunchy top disease in Australia.

Isometric virus-like particles, 18 nm in diameter, have been isolated from banana (Musa spp.) affected by bunchy top disease in Australia. Banana bunchy top disease-associated virus-like particles (BBTV) banded as a single component with buoyant density of 1.28 to 1.29 g/ml in Cs2SO4 and sedimented at about 46S in isokinetic sucrose density gradients. The A260/A280 of purified preparations was about 1.33. A single coat protein of Mr 20,500 was identified with antibodies to BBTV particles from Australia. Single-stranded DNA of about 1 kb as well as ssRNA smaller than 0.45 kb was also associated with the particles. A polyclonal antiserum to BBTV, suitable for use in ELISA, was prepared. Stability and antigenicity of purified BBTV was impaired by storage at pH greater than or equal to 8.5 and freezing at -20 degrees C without protectants. BBTV was detected by double antibody sandwich-ELISA with monoclonal and polyclonal antibodies, in field-infected banana plants, single aphids from an infective colony, and in experimentally aphid-inoculated banana plants. After transmission of BBTV particles by aphids from a banana bunchy top disease-affected to an uninfected banana plant, the disease was induced and BBTV was detected by ELISA in symptomatic leaves only. BBTV isolates from Australia, Taiwan, People's Republic of China, Tonga, Western Samoa and Hawaii were found to be serologically related, which suggests a common aetiology for the disease.

Animals↗
PubMed1991

Helicobacter pylori in children.

The clinical manifestations of Helicobacter pylori (formerly Campylobacter pylori) infection in children overlap with those encountered in adults but there are some important differences. The aim of this review is to describe the range of manifestations of this infection in children, together with means of diagnosis and treatment.

Adolescent↗
PubMed1991

Behçet's disease associated with HIV infection.

The cooccurrence of Behçet's disease and human immunodeficiency virus infection is described in a 33-year-old man who died. If this is not a chance occurrence, then either the constellation of signs that characterize Behçet's disease may be simulated by complications of HIV infection, or else HIV infection causes, or predisposes to, a Behçet's-like illness.

Adult↗