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K J Robson

Publications and source records attributed to K J Robson.

At least 55 records · Page 3Linked to original sources
PubMed1997

The HLA A1-B8 haplotype extends 6 Mb beyond HLA-A: associations between HLA-A, B, F and 15 microsatellite markers.

Allele frequencies for HLA-A, B and F and 15 microsatellite markers located from 100 kb telomeric to HLA-A to 6 Mb telomeric have been determined in a group of 60 blood donors. Linkage disequilibrium analysis revealed significant haplotype associations even after correction for the number of comparisons made. The HLA-A1, B8 haplotype extends as far as D6S276 (6.0 Mb telomeric to HLA-A). It is important to realize that this common haplotype extends beyond the HLA region, especially when evaluating haplotype associations with particular disorders.

Biomarkers↗
PubMed1997

Global prevalence of putative haemochromatosis mutations.

Haemochromatosis is a genetic disease associated with progressive iron overload, and is common among populations of northern European origin. HLA-H is a recently reported candidate gene for this condition. Two mutations have been identified, a substitution of cysteine for tyrosine at amino acid 282 (C282Y, nucleotide 845) and of histidine for aspartate at amino acid 63 (H63D, nucleotide 187). Over 90% of UK haemochromatosis patients are homozygous for the C282Y mutation. We have examined 5956 chromosomes (2978 people) for the presence of HLA-H C282Y and H63D by PCR followed by restriction enzyme analysis. We have found world wide allele frequencies of 1.9% for C282Y and 8.1% for H63D. The highest frequencies were 10% for C282Y in 90 Irish chromosomes and 30.4% for H63D in 56 Basque chromosomes. C282Y was most frequent in northern European populations and absent from 1042 African chromosomes, 484 Asian chromosomes, and 644 Australasian chromosomes. The distribution of the C282Y mutation coincides with that of populations in which haemochromatosis has been reported and is consistent with the theory of a north European origin for the mutation. The H63D polymorphism is more widely distributed and its connection with haemochromatosis remains unclear.

Alleles↗
PubMed1996

Pfmrk, a MO15-related protein kinase from Plasmodium falciparum. Gene cloning, sequence, stage-specific expression and chromosome localization.

Cyclin-dependent kinases (Cdks) play a central role in the regulation of the eukaryotic cell cycle. A novel gene encoding a Cdk-like protein, Pfmrk, has been isolated from the human malaria parasite Plasmodium falciparum. The gene has no introns and comprises an open reading frame encoding a protein of 324 amino acids with a predicted molecular mass of 38 kDa. Database searches revealed a striking similarity to the Cdk subfamily with the highest similarity to human MO15 (Cdk7). The overall sequence of Pfmrk shares 62% similarity and 46% identity with human MO15, in comparison to the 49-58% similarity and 34-43% identity with other human Cdks. Pfmrk contains two unique inserts: one consisting of 5 amino acids just before the cyclin-binding motif and the other composed of 13 amino acids within the T-loop equivalent region. Southern blots of genomic DNA digests and chromosomal separations showed that Pfmrk is a single-copy gene conserved between several parasite strains and is located on chromosome 10. A 2500-nucleotide transcript of this gene is expressed predominantly in the sexual blood stages (gametocytes), suggesting that Pfmrk may be involved in sexual stage development.

Amino Acid Sequence↗
PubMed1996

An experimental investigation of the effect of light-box luminance on the detection of low contrast objects in mammography.

It has been hypothesized that light-box luminance is an important factor in the detection of objects on radiographs. In this work, existing psychophysical data relating to the measurement of visual thresholds at various scene luminance levels are applied to the problem of the observation of radiographs on a light-box. These data suggest that for a given stimulus size, the threshold contrast varies little over several orders of magnitude of scene luminance. In a series of contrast detail experiments performed over a wide range of light-box luminances it has been demonstrated that the detection of low contrast objects on mammographic film is dominated by external noise, that is noise on the film, rather than the internal visual noise of the observer. It is therefore suggested that in mammography it is inappropriate to base recommendations for optimal values of light-box luminance on psychophysical studies of visual noise. It has been shown that commonly used light-box luminances are suitable for viewing mammograms at the higher average optical densities now being recommended, provided that precautions are taken to avoid glare and reflection.

Contrast Sensitivity↗
PubMed1995

Thrombospondin-related adhesive protein (TRAP) of Plasmodium falciparum: expression during sporozoite ontogeny and binding to human hepatocytes.

Plasmodium sporozoites collected from oocysts, haemocoel and salivary glands of the mosquito show profound differences in their biological properties such as motility, ability to induce protective immune response and infectivity for vertebrate host cells. Sporozoites from salivary glands are much more infectious than those from oocysts and haemocoel. Differential expression of proteins, such as the circumsporozoite (CS) protein and the thrombospondin-related adhesive protein (TRAP), implicated in sporozoite recognition and entry into hepatocytes may account for the development of infectivity during ontogeny. We have carried out a series of experiments to: (i) analyse the expression and localization of TRAP in P.falciparum sporozoites during development in the mosquito; and (ii) elucidate the biochemical and adhesive properties of recombinant TRAP. Our data indicate that TRAP is not expressed in oocysts, whereas variable amounts of CS protein are found in this parasite developmental stage. Hemocoel sporozoites display the distinct phenotypes TRAP- CS protein+ and TRAP+ CS protein+ at a frequency of 98.5 and 1.5% respectively. Salivary gland sporozoites are all TRAP+ CS protein+. We also provide experimental evidence showing that recombinant TRAP binds to the basolateral cell membrane of hepatocytes in the Disse's space and that sulfated glycoconjugates function as TRAP ligands on human hepatocytes.

Amino Acid Sequence↗
PubMed1995

New polymorphic microsatellite markers place the haemochromatosis gene telomeric to D6S105.

The haemochromatosis gene (HFE) is linked to both HLA-A and D6S105 on the short arm of chromosome 6 but these markers are separated by approximately 2 Mb of DNA. Most chromosomes carrying HFE have a common haplotype which extends from HLA-A to D6S105 and includes HLA-F. To localise the gene more precisely we have examined 10 microsatellite markers extending over a genetic distance of approximately 5 cM from D6S265 (within 100 kb of HLA-A on the centromeric side) to D6S299 (telomeric). The order of markers is D6S265, HLA-F, D6S258, D6S306, CS3, D6S105, D6S464, CS5, D6S461 and D6S299. We confirm that haemochromatosis appears to originate from a founder mutation which has multiplied in the population through successive generations. This mutation is associated with the haplotype D6S306-5, CS3-3, D6S105-8, D6S464-9 and CS5-4 which is found on approximately 70% of HFE chromosomes. We have applied a new and powerful, likelihood analysis for linkage disequilibrium. The maximum value of lambda (proportion of total possible association between a marker and disease) is 0.74 for marker CS5 (allele 4). A multipoint analysis also gives a maximum likelihood near marker CS5. We conclude that the HFE gene is likely to be located telomeric of D6S105 and close to CS5.

Alleles↗
PubMed1995

The use of a contrast-detail test object in the optimization of optical density in mammography.

In mammography, it is important that the maximum amount of diagnostic information is obtained from each radiograph. One of the factors influencing image quality is the optical density of the film. In this study, a contrast-detail test object was used to establish the optimum optical density value on the basis of signal-to-noise ratio (SNR) for two mammographic film-screen combinations under particular processing conditions. The optimum optical density, taken as being the point at which the threshold contrast is a minimum varied little with detail size. The optimum optical density for the Fuji film-screen combination with Photosol chemistry was found to be higher than that for the Kodak film-screen combination and Kodak processing conditions studied. Although the actual values of optical density are specific to the fixed processing conditions encountered, the technique has more general applicability.

Breast Neoplasms↗
PubMed1995

Assessment of mean glandular dose in mammography.

The routine assessment of patient dose in the National Health Service Breast Screening Programme is performed as part of the quality assurance protocol recommended by the Institute of Physical Sciences in Medicine. The mean glandular dose to a standard breast is deduced from measurement of the air kerma at the entrance surface of a 4 cm Perspex phantom by applying a series of conversion factors. The exposure factors for this measurement are those used clinically. The measured mean glandular dose is then compared with nationally accepted action levels. In some centres the assessment of mean glandular dose using Perspex is supplemented by patient dose surveys. The mean glandular dose to a series of patients attending a breast screening unit may be estimated from a knowledge of the exposure factors and compressed breast thickness, using a knowledge of the X-ray tube output. Measurements made on units in the Northern Region of England and in Scotland using both methods are presented. The implication of these measurements with regard to patient dose surveys in mammography and quality assurance programmes are discussed. An analysis of the uncertainties associated with the measurement techniques is presented.

Breast↗
PubMed1994

Measurements of the frequency distribution of optical density in screening mammography.

A method for obtaining histograms of the frequency of occurrence of optical densities in mammograms has been implemented. Averaged optical density distributions were produced for approximately 50 screening mammograms at each of two centres using different film-screen and processing chemistry combinations. Additional measurements were made on films of two commercially available anthropomorphic breast phantoms. The results show that screening mammograms, on average, exhibit a strongly peaked optical density distribution which corresponds closely to the target optical density measured using the current recommended test protocol for automatic exposure control devices.

Breast Neoplasms↗
PubMed1994

6-Hydroxy-4-sphingenine in human epidermal ceramides.

The solvent-extractable lipids of human epidermal stratum corneum consist predominantly of ceramides. In addition two non-extractable ceramides are chemically bound to the stratum corneum protein. One of the bound ceramides, constituting 50% of the bound lipids, was previously shown to consist of very long chan omega-hydroxyacids in amide linkage with sphingosine. The second bound caramide, which forms 25% of the bound lipids, was shown to contain the same hydroxyacids, but the sphingoid base was neither sphingosine nor phytosphingosine. In the present study, the undefined bound ceramide was shown by NMR and chemical procedures to be the omega-hydroxyacid derivative of a new base, 6-hydroxy-4-sphingenine. In addition, a ceramide previously known to constitute 25% of the extractable human stratum corneum ceramides has been found to contain the same novel sphingoid base, amide-linked to long-chain alpha-hydroxyacids. Finally, a new acylceramide has been isolated and identified that consists of very long chain omega-hydroxyacids in amide linkage with the novel sphingolipid, with fatty acids esterified wit the terminal hydroxyl group of the hydroxyacid.

Adult↗
PubMed1994

Isolation of CA dinucleotide repeats close to D6S105; linkage disequilibrium with haemochromatosis.

The gene for hereditary haemochromatosis (HC) is linked to HLA-A and D6S105 on chromosome 6p. Both markers have also been reported to display linkage disequilibrium with the disease. However, their physical localization relative to one another has not been established. We demonstrate by fluorescent in situ hybridisation that D6S105 lies at least 1-2 Mb telomeric of HLA-A. The haemochromatosis critical region extending from proximal of HLA-A to distal of D6S105 is therefore large. To improve the genetic resolution in this region more highly polymorphic markers are required. We have therefore isolated three novel CA dinucleotide repeats close to D6S105. A linkage disequilibrium study, with two of these microsatellites, in HC patients and controls lends support to the conclusion that D6S105 is a close marker to the haemochromatosis gene.

Alleles↗
PubMed1993

Molecular modelling of malaria calmodulin suggests that it is not a suitable target for novel antimalarials.

The recent cloning and sequencing of many calmodulin genes permits alignment of DNA and protein sequences, as well as structural comparison based on homology modelling. The crystal structure of calmodulin places the four Ca(2+)-binding domains in a dumbbell-like configuration, with a large hydrophobic cleft in each half of the molecule. Calmodulin from Plasmodium falciparum has a high level of sequence identity (89%) with its mammalian counterpart. However, a lower degree of sequence conservation is observed among calmodulins from other lower eukaryotes. Potentially important differences in calmodulin sequences involve amino acids with side-chains forming the hydrophobic clefts as well as in the central helix; these differences could alter interactions with small hydrophobic molecules such as chloroquine and with enzymes modulated by calmodulin. Our modelling studies suggest that neither of the antimalarials examined (chloroquine and quinine) bind tightly to calmodulin. We conclude that the differences between host and parasite calmodulins are insufficient to merit this protein being chosen as a realistic target for antimalarial drug design. By contrast, our sequence comparisons reveal that the fungal calmodulins are significantly divergent from those of higher eukaryotes suggesting that at least in these species, calmodulin might be a target for novel antimycotic drugs.

Amino Acid Sequence↗
PubMed1993

Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum binds specifically to sulfated glycoconjugates and to HepG2 hepatoma cells suggesting a role for this molecule in sporozoite invasion of hepatocytes.

Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum contains an amino acid motif based around the sequence WSPCSVTCG which is also found in region II of the circumsporozoite (CS) proteins of different species of Plasmodium. This amino acid motif confers on the CS protein the ability to bind specifically to sulfated glycoconjugates and to hepatocytes. This suggests that the interaction of CS protein with sulfated glycoconjugates on the surface of the hepatocytes may represent the first molecular event of sporozoite invasion of liver cells. Experimental evidence indicates that TRAP is localized both on the micronemes and on the surface of P. falciparum sporozoites implying that TRAP with its putative sulfated glycoconjugate binding motif may also be involved in recognition and/or entry of hepatocytes by the sporozoite. We show here that different TRAP constructs expressed in Escherichia coli bind to sulfogalactosyl-cerebrosides (sulfatides) and to the surface of HepG2 cells. These interactions are dependent on the presence of the conserved amino acid motif WSPCSVTCG within the sequences of the constructs and are completely inhibited by several sulfated glycoconjugates as well as by suramin, a polysulfonated drug with anti-protozoan activity. Moreover, sporozoite invasion of HepG2 cells is inhibited by antisera raised against these different TRAP constructs and by the presence of low concentrations of suramin. We concluded that TRAP may be one of the parasite encoded molecules in the host-parasite interaction that results in sporozoite invasion of hepatocytes.

Amino Acid Sequence↗
PubMed1993

Sequence diversity in the intron of the calmodulin gene from Plasmodium falciparum.

Sequence variation in the single intron of the calmodulin gene of Plasmodium falciparum has been examined following amplification using the polymerase chain reaction (PCR). The intron has 4 repeating motifs varying in length: 3 of these contain a dinunucleotide repeat, dA-dT, the fourth is a pentameric repeating unit, dA-dT-dA-dT-dT. These DNA polymorphisms can be applied to the study of parasite populations in mixed infections and in strain identification. The function of these repeating motifs is unknown. Computer modelling of the possible intron structures demonstrates that each of these repeating motifs forms individual stem loops such that any changes in repeat number are self-compensatory and do not change the overall intron structure. The implications of this sequence variation are discussed.

Animals↗
PubMed1993

Transcripts of the multidrug resistance genes in chloroquine-sensitive and chloroquine-resistant Plasmodium falciparum.

Homologues of the mammalian multidrug resistance gene have been identified in isolates and clones of Plasmodium falciparum and designated pfmdr1 and pfmdr2. Mutations in pfmdr1 have been associated with chloroquine resistance but confirmation could not be obtained in a genetic cross. We have examined the copy number and expression of pfmdr1 and pfmdr2 in chloroquine-sensitive and -resistant P. falciparum and have found no relationship between the copy number of either gene and chloroquine resistance. However, a marked correlation was seen between levels of mRNA transcribed for each gene and chloroquine resistance. Two transcripts of pfmdr1 were detected, and in the asexual blood cycle an 8 kb transcript appeared first, followed by the appearance of a 7 kb species.

Animals↗
PubMed1993

A family of cation ATPase-like molecules from Plasmodium falciparum.

We report the nucleotide and derived amino acid sequence of the ATPase 1 gene from Plasmodium falciparum. The amino acid sequence shares homology with the family of "P"-type cation translocating ATPases in conserved regions important for nucleotide binding, conformational change, or phosphorylation. The gene, which is present on chromosome 5, has a product longer than any other reported for a P-type ATPase. Interstrain analysis from 12 parasite isolates by the polymerase chain reaction reveals that a 330-bp nucleotide sequence encoding three cytoplasmic regions conserved in cation ATPases (regions a-c) is of constant length. By contrast, another 360-bp sequence which is one of four regions we refer to as "inserts" contains arrays of tandem repeats which show length variation between different parasite isolates. Polymorphism results from differences in the number and types of repeat motif contained in this insert. Inserts are divergent in sequence from other P-type ATPases and share features in common with many malarial antigens. Studies using RNA from the erythrocytic stages of the malarial life cycle suggest that ATPase 1 (including the sequence which encodes tandem repeats) is expressed at the large ring stage of development. Immunolocalization has identified ATPase 1 to be in the region of the parasite plasma membrane and pigment body. These findings suggest a possible model for the genesis of malarial antigens.

Adenosine Triphosphatases↗