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Biomedical subjects

D J Robinson

Publications and source records attributed to D J Robinson.

At least 55 records · Page 3Linked to original sources
PubMed1996

The accumulation of protoporphyrin IX in plaque psoriasis after topical application of 5-aminolevulinic acid indicates a potential for superficial photodynamic therapy.

The success reported for the treatment of superficial skin carcinomas by photodynamic therapy with topical application of the photosensitizer precursor 5-aminolevulinic acid has therapeutic implications for the treatment of other skin disorders. This paper describes the accumulation of the photosensitizing agent protoporphyrin IX in areas of plaque psoriasis by monitoring of the fluorescence emission induced by low-intensity laser excitation at 488 nm. We present results from 15 patients with a total of 42 plaques and show that the characteristic fluorescence emission of protoporphyrin IX increases in intensity within the 6-h period following application of 5-ami-nolevulinic acid, suggesting that there is a potential for superficial photodynamic therapy. The rate of increase and maximum intensity of fluorescence emission was not directly related to the applied quantity of the precursor. The variability of the fluorescence intensity was as great between plaques at different sites on the same patient as between different patients. Also, the effect of plaque occlusion following application appeared insignificant. Although there was only limited enhancement of emission from areas of skin surrounding the plaque, a significant buildup of sensitizer was detected after several days in some areas of psoriasis that received no application.

Administration, Topical↗
PubMed1995

Antigenic analysis of nematode-transmissible and non-transmissible isolates of tobacco rattle tobravirus using monoclonal antibodies.

Murine monoclonal antibodies (MAbs) were produced against a nematode non-transmissible isolate of tobacco rattle tobravirus (TRV-PLB). Two of the MAbs (SCR 80 and 81) did not react with the serologically closely related isolate TRV-PPK20 (both isolates belong to the PRN serotype), which is readily transmissible by nematodes. When further isolates of the PRN serotype were tested, all the non-transmissible ones reacted with these two MAbs, but so also did some of those that are readily transmissible. SCR 80 and 81 were able to trap TRV-PLB particles onto electron microscope grids and reacted with metatopes located near the C terminus of the particle protein. The epitope recognized by SCR 81 was discontinuous. The MAbs that reacted with both TRV-PLB and TRV-PPK20 recognized either continuous cryptotopes (SCR 78) or discontinuous neotopes (SCR 79 and 82).

Amino Acid Sequence↗
PubMed1994

Electroencephalography as an aid in the exclusion of Alzheimer's disease.

OBJECTIVE: To determine the usefulness or otherwise of the awake electroencephalogram (EEG) in the diagnosis of Alzheimer's disease (AD). DESIGN: Prospective collection of one or more awake EEGs in patients diagnosed as having AD or mixed AD and multi-infarct dementia according to current systematic criteria with analysis of those cases confirmed by postmortem examination. Systematized blind interpretation of EEGs. SETTING: Tertiary care practice with both ambulatory and hospitalized patients, ie, neurological department in general hospital and psychogeriatric unit in psychiatric hospital. PATIENTS: A series of 86 subjects with AD and 17 with mixed AD and multi-infarct dementia being those members of a consecutive series on whom postmortem information was available. Awake EEGs in 56 age- and sex-matched control subjects. RESULTS: Seventy-five patients with AD (87.2%) and 13 of the mixed group (76.5%) had abnormal EEGs on first testing, giving a sensitivity of 87.2% for uncomplicated AD. Ultimately, 79 (92%) of 86 patients with AD had abnormal EEGs. Twenty (35%) of 56 EEGs for matching control subjects were abnormal. Moderately abnormal or severely abnormal EEGs were found in 10 (50%) of 20 of the patients with AD of less than 4 year's duration compared with two (4.1%) of 49 of the control subjects, giving a specificity of 95.9% for EEGs with this degree of abnormality. The normal EEG had a negative predictive value of 0.825 with respect to the diagnosis of AD in these populations. CONCLUSIONS: Widespread availability, low cost, and high sensitivity support the use of the awake EEG in the diagnosis of AD.

Aged↗
PubMed1994

Sequences of 10 variants of the satellite-like RNA-3 of groundnut rosette virus.

Ten cDNA clones, representing the satellite-like RNA-3 associated with five different isolates of groundnut rosette virus, were prepared and sequenced. The sequences are at least 87% identical, but differences associated both with symptom differences and with geographical origin were found. The sequences contain up to five potential open reading frames (ORFs), of which three are in the strand that is most abundant in infected plants and two are in the complementary strand. Only one of these ORFs is present in all 10 sequences. Neither the sequence of RNA-3 nor that of its potential translation products had significant similarities to sequences in the databases.

Arachis↗
PubMed1994

Sequences at the ends of RNA-2 of I6, a recombinant tobravirus.

Tobravirus isolate I6 is an anomalous strain of tobacco rattle virus (TRV), which has biological properties typical of TRV, but which is serologically related to pea early-browning virus (PEBV). Its RNA-2 species is a natural recombinant molecule which contains internal sequences, including the particle protein gene, resembling those of PEBV, but with some TRV-like sequences at each end. Sequencing of RNA-2 of isolate I6 showed that, at the 3' end, it is almost identical to RNA-2 of PEBV strain SP5, and TRV-like sequences are limited to the 25 residues that are common to both viruses. However, unlike PEBV RNA-2, I6 RNA-2 can be replicated by TRV RNA-1-coded enzymes. Thus, specific recognition of RNA-2 by TRV replicase seems not to involve sequences at the 3' end. About 275 residues at the 5' end of I6 RNA-2 are similar to those in RNA-2 of TRV strain TCM, and are jointed to the PEBV-SP5-like subgenomic promoter and coding sequences by about 100 residues of uncertain origin. These results support the idea that I6 RNA-2 originated by recombination between TRV and PEBV RNA-2 molecules, although the exact position of the junction and therefore the mechanism involved are unclear.

Base Sequence↗
PubMed1993

Nucleotide sequence evidence for the occurrence of three distinct whitefly-transmitted geminiviruses in cassava.

The complete nucleotide sequence of the DNA of Indian cassava mosaic virus (ICMV) and a key part of that of a group B isolate of African cassava mosaic virus from Malawi (ACMV-M) were determined and compared at the nucleotide and encoded amino acid levels with the published sequences of an ACMV group A isolate (ACMV-K) and other whitefly-transmitted gemini-viruses (WTGs). The DNA of ICMV consists of two circular single-stranded molecules, DNA-A [2815 nucleotides (nt)] and DNA-B (2645 nt), which differ substantially in sequence from the genome components of ACMV-K (DNA-A 70%, DNA-B 47% sequence identity) and other WTGs. ICMV DNA-A contains eight open reading frames (ORFs) encoding proteins of > 100 amino acid residues, of which four ORFs (one genome sense, three complementary sense) are comparable to those of other WTGs. DNA-B contains one ORF in each sense, as in other WTGs. None of the putative viral proteins are more similar in amino acid sequence to the proteins of ACMV-K than to those of another WTG. The coat protein of ACMV-M is more like that of tomato yellow leaf curl virus from Sardinia (86% sequence identity) than those of ICMV or ACMV-K. The intergenic regions of ACMV-K, ACMV-M and ICMV DNAs differ in size, and largely in sequence, except for two 30 to 40 nt sequences which are also conserved in other WTGs and can form stem-loop structures. The intergenic region of ICMV DNA contains three copies of a 41 nt sequence, and that of ACMV-M DNA contains an imperfect repeat of a 34 nt sequence which resembles the repeated sequence in ICMV DNA. The differences between ACMV-K, ACMV-M and ICMV are considered great enough to justify their separation as isolates of three distinct WTGs: African cassava mosaic virus, East African cassava mosaic virus and Indian cassava mosaic virus.

Amino Acid Sequence↗
PubMed1993

Susceptibility of transgenic tobacco plants expressing tobacco rattle virus coat protein to nematode-transmitted and mechanically inoculated tobacco rattle virus.

Transgenic Samsun NN tobacco plants expressing the coat protein of tobacco rattle virus were exposed to mechanical leaf inoculation with tobacco rattle virus and to viruliferous trichodorid vector nematodes. Whereas plants were resistant to mechanical inoculation the vector nematodes successfully transmitted tobacco rattle virus to the roots as well as to the leaves of these plants. It is suggested that transgenic resistance is overcome either because vector nematodes inject relatively large numbers of virus particles into a cell or because they inject destabilized particles. The results indicate that coat protein-mediated resistance is unlikely to be of value for controlling tobacco rattle virus in field crops.

Animals↗
PubMed1993

RNA-2 of tobacco rattle virus encodes the determinants of transmissibility by trichodorid vector nematodes.

Pseudorecombinant isolates produced from an efficiently transmitted and a non-transmitted isolate of tobacco rattle tobravirus were tested for their transmissibility by trichodorid vector nematodes. Isolates with RNA-2 originating from the non-transmissible isolate were not transmitted by the vector, whereas isolates with RNA-2 originating from the efficiently transmitted isolate were transmitted efficiently. It is therefore concluded that the factor determining vector transmissibility is located on the RNA-2 genome segment of TRV. Because the serological properties of the isolates also correlate with transmissibility, it is likely that the virus particle protein is involved in the transmission process.

Animals↗
PubMed1992

Detection of tobacco rattle virus by reverse transcription and polymerase chain reaction.

An assay involving reverse transcription followed by polymerase chain reaction specifically detected tobacco rattle virus RNA in extracts of infected plants. It detected a wide range of serological variants and typical non-particle producing NM-type isolates. The sensitivity of the method was sufficient to detect TRV RNA in 10 ng total nucleic acid from an infected plant, or in a relatively crude nucleic acid preparation from 60 micrograms infected leaf tissue.

Base Sequence↗
PubMed1992

The nucleotide sequence of RNA-2 of raspberry ringspot nepovirus.

The nucleotide sequence of raspberry ringspot nepovirus (RRV) RNA-2 consists of 3928 nucleotides and a poly(A) tract at the 3' end. RNA-2 contains one open reading frame which encodes a polypeptide of M(r) 123508 (123K). Edman degradation located the N terminus of the coat protein 514 residues from the C-terminal end of the 123K protein, which suggests that the coat protein is released from the polyprotein by cleavage of a C-A bond. The RRV coat protein has some sequence similarities with the coat proteins of other nepoviruses, but is no more like any one nepovirus than another. In contrast, the portion of the 123K protein to the N-terminal side of the coat protein is similar in sequence to the corresponding parts of the polyproteins of tomato black ring and grapevine chrome mosaic nepoviruses, though not to those of other nepoviruses.

Amino Acid Sequence↗
PubMed1991

Secondary structures of narcissus mosaic virus coat protein.

Circular dichroism (CD) measurements on the coat protein of narcissus mosaic virus particles show that the dominant secondary structure is the alpha-helix, with a 45 (+/- 2)% content. The beta-sheet content is much lower at 5 (+/- 3)%. Both values are essentially the same as those found in potato virus X coat protein. The CD results are used to assess the results of secondary structure prediction methods.

Capsid↗
PubMed1991

Nuclear location of the 16K non-structural protein of tobacco rattle virus.

An antiserum, elicited by a synthetic peptide coupled to bovine serum albumin, reacted specifically with the non-structural 16K protein of tobacco rattle virus. The protein was detected in extracts of systemically infected Nicotiana clevelandii leaves, but only in those made with the aid of SDS, urea and 2-mercaptoethanol. Immunogold labelling of ultrathin sections showed that the protein was mainly associated with nuclei, but was also present in the cytoplasm. These observations suggest that the 16K protein binds to macromolecular components of infected cells, especially in nuclei, but do not clarify its function.

Amino Acid Sequence↗
PubMed1989

Nucleotide sequence of potato leafroll luteovirus RNA.

A sequence of 5987 nucleotides is reported for the RNA of potato leafroll luteovirus (PLRV). The sequence contains six large open reading frames, and non-coding regions of 174 nucleotides at the 5' end, 141 nucleotides at the 3' end and 197 nucleotides between two large blocks of coding sequences. The 5' coding region encodes two polypeptides of 28,000 (28K) and 70K which overlap in different reading frames and circumstantial evidence suggests that the third open reading frame in the 5' block is translated by frameshift readthrough near the end of the 70K polypeptide to give a 118K polypeptide. The C-terminal part of the 118K protein contains the consensus sequence for RNA-dependent RNA polymerases. In vitro translation of PLRV RNA resulted in the synthesis mainly of 28K and 70K polypeptides and the largest product made was about 125K; these sizes are similar to those predicted for the translation products of the 5' block of coding sequence. The 3' block of coding sequence codes for three polypeptides: a 23K coat protein, a 17K polypeptide which is encoded in a different frame, and a 53K polypeptide which immediately follows the coat protein coding sequence, and is in the same reading frame. Circumstantial evidence suggests that the 53K polypeptide is translated by readthrough of the amber termination codon of the coat protein gene. The amino acid sequences encoded by the 3' block of coding sequence show many similarities with analogous polypeptides translated from the nucleotide sequences of RNA of barley yellow dwarf virus, PAV strain (BYDV) and, in particular, beet western yellows virus (BWYV). The 118K polypeptide has some similarities with the putative polymerase of southern bean mosaic virus and much more extensive similarities with the corresponding BWYV polypeptide but almost none with that of BYDV. In contrast, the amino acid sequence of the 28K polypeptide is not like that of proteins of the other luteoviruses or of viruses in other groups. The nucleotide sequences reported will appear in the EMBL, GenBank and DDBJ databases under the accession number X14600.

Amino Acid Sequence↗
PubMed1988

Molecular variation in vector-borne plant viruses: epidemiological significance.

Patterns of variation are examined in four groups of plant viruses, with special reference to their particle proteins and to changes in vector transmissibility and specificity. In the nepoviruses and potyviruses, non-circulative transmission, by nematodes and aphids respectively, seems dependent on structural features on the surface of the virus particles. The N-terminal part of the particle protein may play the key role in potyviruses. Similarly in the luteoviruses, and possibly in the geminiviruses, specificity of circulative transmission by aphids, whiteflies and leafhoppers is linked to the antigenic specificity of the virus particles. Among naturally occurring isolates of the same virus, variation seems often to be discontinuous, and is predominantly of two sorts. Minor variations, characterized by loss of an epitope or substitutions of a few amino acids, can be associated with loss of transmissibility in luteoviruses and potyviruses, or have no effect. Major variations are associated with differences in vector specificity and seem likely to involve radical genetic changes that have evolved over long periods. The adaptation of virus particle proteins for transmission by vectors probably results in conservation of the genes that encode them, and in greater conservation of some parts of these genes than of others.

Amino Acid Sequence↗
PubMed1987

The complete nucleotide sequence of tobacco rattle virus RNA-1.

The complete nucleotide sequence of tobacco rattle virus (TRV) strain SYM RNA-1 was determined from a series of overlapping cDNA clones. cDNA prepared by primer extension was used to determine the exact 5' terminus. The RNA sequence was 6791 nucleotides in length and contained four open reading frames (ORFs). The ORF nearest the 5' terminus coded for a polypeptide of predicted mol. wt. 134,000 (134K) and terminated at an opal (UGA) stop codon. Readthrough of this stop codon would result in the production of a protein of 194K. The gene for a 29K polypeptide started one base beyond the 194K stop codon and, in turn, was followed by the gene for a 16K protein at the 3' end of RNA-1. Amino acid comparisons of the 194K protein with the putative replicase of tobacco mosaic virus showed three regions of strong homology, suggesting that the 134K and 194K proteins were similarly involved in virus replication. The 5' terminal sequences of both genome RNA species of TRV strains ORY, N5 and PRN together with that of SYM RNA-2 were also determined. Alignments of these sequences showed that there was a 22 base repeated sequence close to the 5' terminus in all these RNA species. It was also shown that the 5' terminus of RNA-1 was homologous with the same region in RNA-2.

Amino Acid Sequence↗
PubMed1986

A comparison of the in vivo uptake and metabolism of 3H-oestrone and 3H-oestradiol by normal breast and breast tumour tissues in post-menopausal women.

After infusion of 3H-oestradiol or 3H-oestrone into post-menopausal women with breast cancer, there was a significant uptake of both steroids by breast tumour and normal tissue. The proportion of oestrogen present in tumour tissue as 3H-oestradiol after infusion of 3H-oestradiol (89.4 +/- 3.5%, mean +/- SD, n = 4) was significantly higher (p less than 0.001) than in normal breast tissue (72.8 +/- 3.3%) obtained from the same women. Similarly, after infusion of 3H-oestrone, the proportion of oestrogen present as 3H-oestradiol (48.4 +/- 14.4%) was significantly higher than in normal breast tissue (19.1 +/- 6.4%). These results suggest that conversion of oestrone to oestradiol is enhanced in breast tumour tissue with little metabolism of oestradiol. This would account for the higher concentrations of oestradiol reported in breast tumour tissue in the presence of increased oestradiol 17 beta-hydroxysteroid dehydrogenase activity.

Aged↗