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Biomedical subjects

R Harvey

Publications and source records attributed to R Harvey.

At least 91 records · Page 5Linked to original sources

Cellular RNA is not degraded in interferon-treated HeLa cells after poliovirus infection.

A drastic inhibition of protein synthesis occurs in HeLa cells treated with human lymphoblastoid interferon and infected with poliovirus. At the time when this inhibition has been established no degradation of 32P-labelled ribosomal RNA can be detected. Isolation of the mRNAs from poliovirus-infected cells plus or minus interferon treatment, followed by translation in a reticulocyte lysate indicates that cellular mRNAs remain active. These results suggest that gross degradation of cellular RNA does not occur in interferon-treated poliovirus-infected HeLa cells and that a non-specific nuclease induced by 2'-5' A is not responsible for the inhibition of protein synthesis observed.

Cell Transformation, Viral↗

An antibody to a synthetic peptide that recognises SV40 small-t antigen.

A peptide Tyr.Arg.Asp.Leu.Lys.Leu corresponding to the carboxy-terminal six amino acids of small-t antigen predicted from the DNA sequence of SV40 was synthesised, coupled to bovine serum albumin and to ovalbumin and used to raise antibody in rabbits. The sera obtained immunoprecipitated [125I]peptide. It also recognised SV40 small-t that was synthesised in vitro from SV40 mRNA or extracted from SV40 infected monkey cells. The immunoprecipitation of small-t was inhibited by added peptide. To demonstrate that the determinant was present at the carboxy-terminal end of the molecule, truncated versions of small-t coded for by 0.54-0.59 deletion mutants were tested. dl 890 small-t, which contains an in-phase deletion removing nine amino acids but leaving the carboxy-terminal sequences intact, was recognised by the antipeptide serum. By contrast dl 885 small-t, which has an out-of-phase deletion leading to an altered carboxy terminus coded in an alternative reading frame, was not recognised. The data confirm the location and specificity of the determinant recognised on small-t by the antipeptide serum.

Animals↗

Structure and biochemical functions of four simian virus 40 truncated large-T antigens.

The structure of four abnormal T antigens which are present in different simian virus 40 (SV40)-transformed mouse cell lines was studied by tryptic peptide mapping, partial proteolysis fingerprinting, immunoprecipitation with monoclonal antibodies, and in vitro translation. The results obtained allowed us to deduce that these proteins, which have apparent molecular weights of 15,000, 22,000, 33,000 and 45,000, are truncated forms of large-T antigen extending to different amounts into the amino acid sequences unique to large-T. The proteins are all phosphorylated, probably at a site between amino acids 106 and 123. The mRNAs coding for the proteins probably contain the normal large-T splice but are shorter than the normal transcripts of the SV40 early region. The truncated large-Ts were tested for the ability to bind to double-stranded DNA-cellulose. This showed that the 33,000- and 45,000-molecular-weight polypeptides contained sequences sufficient for binding under the conditions used, whereas the 15,000- and 22,000-molecular-weight forms did not. Together with published data, this allows the tentative mapping of a region of SV40 large-T between amino acids 109 and 272 that is necessary and may be sufficient for the binding to double-stranded DNA-cellulose in vitro. None of the truncated large-T species formed a stable complex with the host cell protein referred to as nonviral T-antigen or p53, suggesting that the carboxy-terminal sequences of large-T are necessary for complex formation.

Amino Acid Sequence↗

A survey of practising psychiatrists' views on the treatment of agoraphobia.

The views of practising psychiatrists on the treatment of agoraphobia were investigated as part of a Quality Assurance Project. A one in six random sample of psychiatrists was mailed a questionnaire. This asked for treatment recommendations for each of three case descriptions of patients with agoraphobia and respondents were asked to choose three treatments from a predetermined list of treatments. Eighty-six percent of the sample responded. Behavioural psychotherapy was recommended most often, usually in conjunction with psychodynamic psychotherapy. Recommendations varied across the three cases and were also marginally affected by the format of the question used to elicit them.

Adult↗

Vertebrate histone genes: nucleotide sequence of a chicken H2A gene and regulatory flanking sequences.

The DNA sequence of a chicken genomal fragment containing a histone H2A gene has been determined. It contains extensive 5' and 3' flanking regions and encodes a protein identical in sequence to the histone H2A protein isolated from chicken erythrocytes. In the 5' flanking region, a possible "TATA box" and three possible "cap sites" can be recognised upstream from the initiation codon. To the 5' side of the "TATA box" is found an unusual sequence of 21 A's interrupted by a central G residue. It occupies the same relative position as the P. miliaris H2A gene-specific 5' dyad symmetry sequence and the "CCAAT box" seen in other eukaryotic polymerase II genes but is clearly different from both. A significant feature of the 3' non-coding region is the presence of a 23 base-pair sequence that is nearly identical to a conserved region found in sea urchin histone genes. The coding region is extremely GC rich, with strong selection for these bases in the third position of codons. Not a single coding triplet ends in U. No intervening sequences were found in this gene.

Animals↗

Does psychotherapy benefit neurotic patients.

Smith, Glass, and Miller examined the benefits of psychotherapy in 475 controlled studies. Their data were reexamined, and only studies of patients coded as having neuroses, true phobias, and emotional-somatic complaints and who had sought treatment were included in the present analysis. The results of 81 controlled trials were integrated statistically using the metaanalytic technique. The findings provide further evidence on the efficacy of psychotherapy. The condition of the typical patient after treatment was better than that of 77% of untreated controls measured at the same time. The rate of relapse in the first two years was small. The behavior and psychodynamic verbal therapies appeared to be superior to other therapies. The relation between severity of illness and choice of therapy is unknown. This factor could account for some of the differential effects but does not vitiate the main finding.

Adolescent↗

Regression to the mean in pretreatment measures of stuttering.

Pre-post treatment evaluation designs are common in stuttering research. Their propriety depends on the assumption that spontaneous remission is not likely. There are six studies in the literature in which stutterers have been measured on two occasions some months apart. In all studies there was a trend to less stuttering on the second assessment, but in no study was the difference between scores significant. One hundred and thirty-two stutterers awaiting treatment were assessed when they were first seen and then at the beginning of treatment 1--23 months later. There was a small but significant improvement between the two assessments. The size of the improvement was comparable to those reported in the six published studies. This spontaneous improvement occurs mainly in the three months following the first assessment, and there is little change thereafter. It is concluded that pre-post studies of subjects who waited more than three months for treatment are valid and that the observed improvement can be due solely to the effects of treatment. Studies that assess improvement from the time subjects are first seen should allow for spontaneous remission to determine the improvement due to treatment.

Adult↗

Large and small tumor antigens from simian virus 40 have identical amino termini mapping at 0.65 map units.

Large and small tumor (T)antigens of simian virus 40 were synthesized in vitro with L-cell extracts that had been treated by the method of Palmiter to prevent amino-terminal acetylation of nascent proteins. Partial amino-terminal amino acid sequences of both forms of T-antigen were determined and found to be identical. Methionine residues were located at positions 1 and 14, a lysine residue at position 3, and leucine residues at positions 5, 11, 13,16, 17, and 19. These amino acid sequence data match perfectly the amino acid sequence predicted from a sequence of nucleotides in the E strand of simian virus 40 DNA which begins near the junction between HindII/III fragments A and C at about 0.65 map units. This strongly suggests that the sequence coding for the amino terminus of both proteins is located at this position. Furthermore, the data are consistent with a model for the synthesis of both forms of T-antigen that predicts that (i) small T-antigen is coded for by a sequence of nucleotides from the 5' end of the early region and (ii) large T-antigen is coded for by nucleotide sequences from two noncontiguous regions of simian virus 40 DNA.

Amino Acid Sequence↗

Polyoma virus complementary RNA directs the in vitro synthesis of capsid proteins VP1 and VP2.

Polyoma virus complementary RNA, synthesized in vitro by using highly purified Escherichia coli RNA polymerase and nondefective form I polyoma DNA, was translated in a wheat germ cell-free system. Polypeptides were synthesized that comigrated on sodium dodecyl sulfate-polyacrylamide gels with the polyoma capsid proteins VP1 and VP2, although most of the cell-free products were of smaller molecular weights. The VP1-size protein specifically immunoprecipitated with anti-polyoma virus serum, and upon digestion by trypsin yielded [35S]methionine-labeled tryptic peptides that co-chromatographed with the [3H]methionine-labeled tryptic peptides of virion-derived VP1 on both cation-exchange and anion-exchange resins. The VP2-size in vitro product contained all the virion VP2 methionine-labeled tryptic peptides, as shown by cation- and anion-exchange chromatography and two-dimensional fingerprinting on cellulose. We conclude that full-length polyoma VP1 and VP2 are synthesized in response to complementary RNA and consequently that the viral capsid proteins VP1, VP2, and VP3 are entirely virus coded.

Capsid↗

Cell-free synthesis of simian virus 40 T-antigens.

Polyacrylamide gel electrophoresis and tryptic peptide fingerprint analysis of the proteins made in a cell-free system derived from L-cells and immunoprecipitated with simian virus 40 (SV40) anti-T serum demonstrated that both SV40 large-T and small-T antigens are synthesized in vitro in response to mRNA isolated from productively infected CV1 CELLS. Sucrose density centrifugation in gradients containing 85% formamide showed that the mRNA's for both forms of T-antigen sediment at about 17.5S, with the mRNA for small-t sedimenting marginally, but reproducibly, ahead of the mRNA for large-T. Hybridization experiments using restriction endonuclease fragments Hae III-E and Hind II/III-B showed that all fractions active in the cell-free synthesis of both forms of T-antigen hybridized equally to both fragments. This suggests that the mRNA's for SV40 T-antigens are at least partly virus coded and that the bulk of the early SV40 mRNA contains sequence information from both ends of the early region. The data are consistent with the suggestion that the large-T mRNA is spliced. SV40 complementary RNA (the product of transcription of SV40 DNA using Escherichia coli RNA polymerase) was also translated in the L-cell system and gave two families of polypeptides which specifically immunoprecipitate with anti-T serum. One family (the small-t family) includes a polypeptide indistinguishable by gel electrophoresis and tryptic peptide fingerprinting from small-t isolated from cells. The other family (the 60K family) has a major component with molecular weight approximately 60,000 and includes other polypeptides with molecular weights ranging from approximately 14,000 to about 70,000. The 60K family has petides in common with large-T but not with small-T. Together, the peptides of the small-t and 60K families account for virtually all of the methionine peptides of SV40 large-T. We conclude from these results (i) that small-t is probably entirely, and large-T at least predominantly, virus coded; (ii) that the small-t and 60K families represent the translation products of two different portions of the early region of SV40 DNA (approximately 0.65 to 0.55 map units and 0.54 to 0.17 map units); and (iii) that although most, if not all, of the large-T and small-t peptides are present in the cell-free product, some feature of sequence arrangement of SV40 complementary RNA prevents the translation of full-length large-T and results instead in the synthesis of fragments. We suggest that the absence of a splice in the complementary RNA is responsible for this result.

Antigens, Viral↗

Grass pollen content of the air in the Bristol Channel region in 1976.

The grass pollen content of the air at four sites bordering the Bristol Channel (at Bristol, Cardiff, Penarth and Penmaen) was studied. The trap at Penmaen was situated at an official climatological station which provided the meteorological data used in the surveys. Although at certain times large differences could be seen between the concentrations of grass pollen at the different sites, overall the findings for the four sites were similar. Changes in the atmospheric pollen concentration related to variations in meteorological conditions. At the peak of the season high concentrations of grass pollen persisted throughout the night.

Atmosphere↗