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Biomedical subjects

I D Walker

Publications and source records attributed to I D Walker.

At least 181 records · Page 10Linked to original sources

Induction of delayed-type hypersensitivity to azobenzenearsonate by a monoclonal anti-idiotype antibody.

Azobenzenearsonate (ABA)-specific sensitivity was induced in A/J mice by injecting a monoclonal anti-idiotype reagent, 14A, directed against a determinant present on a minor subpopulation of immunoglobulin molecules within the anti-ABA antibodies of A/J mice. Sensitivity was transferrable by purified T cells and this was abrogated by treating the cells with 14A, rabbit anti-mouse immunoglobulin and complement, not by treatment with only the last two reagents. The transfer was restricted by the K-end of the major histocompatibility complex.

Animals↗

A double blind study of prostacyclin in cardiopulmonary bypass surgery.

In a double-blind placebo-controlled trial of 48 patients requiring open heart surgery, prostacyclin (PGI2) was infused in a dose of 20 ng/kg/min throughout cardiopulmonary bypass. When compared with the placebo group, the patients given active PGI2 were found to have significantly higher platelet counts from 30 min after commencement of bypass and in the immediate post-operative period, and to have significantly less elevation of the platelet secretory proteins, beta thromboglobulin and platelet factor 4 during bypass. The mean weight increase in the arterial line filters was significantly greater in the placebo-treated patients than in the PGI2 group. It is suggested that infused PGI2 decreases platelet activation during cardiopulmonary bypass and that further studies are required to establish its clinical value in this situation.

Adult↗

Monoclonal anti-azobenzenearsonate antibodies expressing the cross-reactive idiotype: immunochemical studies show that all idiotypic determinants reside on a single molecule.

Cell lines that secreted antibodies to the hapten azobenzenearsonate (ABA) were established by hybridization of immune A/J spleen cells to the non-secreting myeloma, NS-1. Solid-phase radioimmunoassays (RIA) were developed for rapid screening of hybridoma supernatants to detect antibodies to ABA and to detect antibodies bearing the ABA cross-reactive idiotype (CRI). Hybrid clones secreting both CRI+ and CRI-- anti-ABA antibodies were obtained. The supernatant from one clone (7-13) strongly inhibited binding of iodinated anti-idiotype serum in a competitive RIA. This clone expressing the CRI produced immunoglobulin of the IgG2a subclass. Solid-phase absorption of anti-idiotype serum followed by competitive radioimmunoassay analyses revealed that all the idiotypic determinants recognized by anti-idiotype serum reside on this monoclonal antibody.

Animals↗

mu Polypeptide phenotypes of lymphoid and myeloid cell lines containing RNA transcripts of the immunoglobulin C mu gene.

We have examined whether polyadenylated-RNA molecules encoded by the immunoglobulin C mu gene (C mu RNA) that have recently been shown to occur in mouse T lymphoma, myeloid tumor, and pre-B lymphoma cell lines are translated. Three T lymphoma, 1 myeloid tumor, and 4 Abelson lymphoma cell lines, all known to contain C mu RNA molecules, were examined for evidence of synthesis of mu polypeptides using radioimmunoassay, biosynthetic peptides were detected in any of the T lymphoma or myeloid cell lines, although mu-chains were readily detected in 2 B lymphoma lines known to contain small amounts of mu mRNA. Detection failure is unlikely to be the result of assay insensitivity and was not caused by lack of an appropriate antiserum reactivity or a requirement for mu-chains to be associated with immunoglobulin light chains. Proteolytic activity that might degrade newly synthesized mu polypeptides in these cell lines was not detected. In 3 of the 4 Abelson lymphoma cell lines containing C mu RNA, corresponding polypeptides were absent, although in the 4th they were easily detected, indicating that transcription of the C mu gene is not the sole determinant of whether a cell will synthesize the corresponding polypeptide. The parallel between the C mu RNA+, mu polypeptide- Abelson cell lines and the T lymphoma cell lines suggests that the onset of C mu transcription may occur in both T and B lineages before translational competence is acquired.

Animals↗

Immunoglobulin Cmu RNA in T lymphoma cells is not translated.

It is widely believed that immunoglobulin genes might encode at least part of the receptor for antigen on the T lymphocyte. Evidence supporting this comes from the effects of anti-immunoglobulin idiotype antibodies on cellular immune networks and from the presence of idiotypes on immunologically active factors from T cells. Detailed molecular characterization of the receptors, however, has been seriously hampered by the lack of a suitable cellular source from which it might be isolated. The recent demonstration of Kemp et al. that thymocytes and certain cultured lines of mouse T lymphoma cells contain polyadenylated RNA molecules encoded by the immunoglobulin Cmu gene (Cmu RNA) prompted us to identify the corresponding protein molecules in those cells. As the haploid mouse genome contains a single Cmu gene, any polypeptide encoded by this gene should react with at least some of the antibodies present in rabbit anti-mouse IgM antiserum. In this letter we report that a number of T lymphoma lines, regardless of whether they contain Cmu RNA, synthesize no detectable mu polypeptides.

Animals↗

Allelic forms of beta 2-microglobulin in the mouse.

Spleen cells from BALB/c and C57BL/6 mice were cultured separately or together, and the biosynthetically labeled supernates were examined by two-dimensional polyacrylamide gel electrophoresis. Although there were no major labeled proteins in the mixed group that were not present in the separate cultures, there was a major low-molecular-weight protein that differed in charge in the two strains. This protein was identified as beta 2-microglobulin; it could be labeled with 125I on the cell surface by using the lactoperoxidase technique, was noncovalently attached to the H-2K molecule, and had the expected size and charge when compared with human beta 2-microglobulin. Both acidic and basic forms were present in (BALB/c X C57BL/6) F1 hybrids, suggesting codominant expression, although allelic exclusion was not ruled out. Either parental form could combine with one parental form of the H-2K molecule. The beta 2-microglobulin gene does not appear to be closely linked to either the H-2 or th immunoglobulin heavy-chain complexes. It is proposed that beta 2-microglobulin is an "effector subunit" of histocompatibility antigens and that its physiological role is to interact with a specific killing structure on the surface of cytolytic T lymphocytes and thereby initiate cell destruction.

Alleles↗

Maintenance control of oral anticoagulant therapy by a chromogenic substrate assay for factor X.

An amidolytic assay employing the chromogenic substrate S 2337 (Kabi Diagnostica) was used to assay factor X in 35 healthy controls and in 100 outpatients receiving oral anticoagulant therapy. This method correlated well with a coagulation assay of factor X in the control group (r = 0.88). When compared with two routine tests for the control of anticoagulant theray (Thrombotest and prothrombin ratio) good correlation was obtained between the methods, r = 0.84 and r = -0.74 respectively. These results suggest that a chromogenic substrate assay for factor X might be a suitable method for the maintenance control of oral anticoagulant therapy.

Adult↗

Breast or bottle.

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Bottle Feeding↗

The keratin chains of avian scale tissue. Sequence heterogeneity and the number of scale keratin genes.

The three major proteins of chick scale keratin were isolated as their S-carboxymethylated derivatives and shown to be similar or identical in molecular weight by gel filtration but to be distinct by amino acid analysis and gel electrophoresis. The major amino-terminal sequence of scale keratin chains was determined and shown to be highly homologous to the corresponding region of feather keratin chains. The carboxyl-terminal peptides of the three scale keratin fractions differed in sequence but were all homologous to the carboxyl-terminal segment of feather keratin chains. The pronounced concentration of cysteine residues at the amino-terminal and carboxyl-terminal segments suggested a similar role for these regions in both scale and feather keratin chains, namely to provide a disulphide-linked matrix to maintain the organisation of fibrils which arise from the internal hydrophobic segments of both types of chain. Analysis of a large hydrophobic segment from each of the three isolated protein fractions revealed that each was composed largely of repeating tripeptide units of the type Gly-Gly-X (where X = Phe, Leu or Tyr). At a few positions in each hydrophobic peptide, microheterogeneity was apparent in the sequences indicating that each isolated protein fraction was composed of at least three different chains each encoded by a different gene. A minimum of nine keratin genes are therefore expressed in scale tissue.

Amino Acid Sequence↗

Photoreceptor protein from the purple membrane of Halobacterium halobium. Molecular weight and retinal binding site.

The apparent molecular weight of the purple membrane protein of Halobacterium halobium was found to be 20 000 by sodium dodecyl sulfate gel electrophoresis and by gel filtration in sodium dodecyl sulfate. However, the molecular weight value determined by gel filtration in 6 M guanidine was 28 000. To resolve this discrepancy, methods insensitive to or independent of the conformation of the protein were used to estimate the molecular weight. Analytical ultracentrifugation of the sodium dodecyl sulfate-protein complex, peptide mapping, and amino acid analysis all gave values of 25 000 +/- 1000, a figure in agreement with a recent x-ray study. Borohydride reduction was used to attach the retinal cofactor covalently to a lysine residue. After digestion with thermolysin, peptide maps were prepared of the protein labeled at lysine residues with [14C] succinic anhydride both before and after reduction. Comparison of the maps showed one radioactive peptide with changed mobility. This peptide was isolated and shown to have the sequence Val-Ser-Asp-Pro-Asp-Lys-Lys with only one of the two lysine residues alkylated. Solid-phase sequencing showed the succinyl group to be at position 6 and hence the retinal group to be at position 7. It was possible that a small amount of retinal was also bound to Lys-6. There was no apparent homology with the corresponding peptide of vertebrate rhodopsin. No evidence of chain heterogeneity was found by radiochemical peptide mapping and sequence analysis of peptides containing lysine residues indicating that all protein chains of purple membrane are very similar or identical.

Amino Acid Sequence↗

The subunits of rabbit-muscle phosphofructokinase. A search for sequence repetition.

Rabbit muscle phosphofructokinase uniformly carboxymethylated with iodo[2-14C]acetate consists of subunits with a molecular weight of 80 000 +/- 5000. The subunit polypeptide chain contains 16 and 52 residues respectively of cysteine and arginine and, contrary to previous results, peptide mapping experiments gave no indication that phosphofructokinase chains yield fewer than the expected numbers of cysteine and arginine containing peptides. To test further for the possible occurrence of repeat sequences within a single subunit chain, cysteine-containing peptides were isolated and sequenced from tryptic and thermolytic digests of s-[2-14C]carboxymethylated phosphofructokinase. In all, 15 different cysteine sequences (comprising a total of 104 residues) were identified, showing that not more than one of an expected 16 cysteine-containing sequences is repeated, and that the subunits of phosphofructokinase are of unique sequence along their entire length. The near quantitative isolation of several cysteine-containing peptides shows further that all subunits are of similar if not identical sequence.

Amino Acid Sequence↗

Plasma fibrinolytic activity following oral anabolic steroid therapy.

Six anabolic steroids were assessed for their ability to enhance plasma fibrinolytic activity in males with ischaemic heart disease. Five 17alpha-alkylated steroids (Ethyloestrenol, Norethandrolone, Methandienone, Methylandrostenediol and Oxymetholone) were examined and all produced a significant increase in plasma plasminogen activator as measured by the euglobulin lysis time. The only non-17alpha-alkylated steroid studied (Methenolone acetate) failed to enhance fibrinolysis. The 17alpha-alkylated steroids studied all deserve more detailed evaluation of their long term effects on plasma fibrinolytic activity.

Adult↗

Effect of anabolic steroids on plasma antithrombin III. alpha2 macroglobulin and alpha1 antitrypsin levels.

The effect of seven different anabolic steroids (Ethyloestrenol, Methenolone acetate, Norethandrolone, Methylandrostenediol, Oxymetholone, Methandienone, and Stanozolol) on three alpha-globulin antiprotease inhibitors of thrombin and plasmin was studied in men with ischaemic heart disease. In distinct contrast to the oral contraceptives, five of the six 17-alpha-alkylated anabolic steroids studied produced increased plasma Antithrombin III levels and five produced decreased levels of plasma alpha2-macroglobulin. The effect on plasma alpha1-antitrypsin levels was less clear-cut but three of the steroids examined produced significantly elevated levels. The increased plasma fibrinolytic activity which the 17-alpha-alkylated anabolic steroids induce is therefore unlikely to be secondary to disseminated intravascular coagulation.

Adult↗