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L Henderson

Publications and source records attributed to L Henderson.

At least 109 records · Page 6Linked to original sources

Expression of a rat brain creatine kinase-beta-galactosidase fusion protein in Escherichia coli and derivation of the complete amino acid sequence of rat brain creatine kinase.

Expression of a rat brain creatine kinase (CKB)/beta-galactosidase fusion protein in Escherichia coli has allowed isolation of a rat CKB cDNA clone by direct antibody screening of a rat brain gamma gt11 expression library. This clone is 1416 bp long and includes 202 bp of 3'-untranslated region and 29 bp of 5'-untranslated region. The coding sequence of this clone has enabled us to deduce the complete amino acid (aa) sequence of rat CKB protein.

Amino Acid Sequence↗

Interleukin 3.

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20-Hydroxysteroid Dehydrogenases↗

Isolation and sequence analysis of cDNA clones coding for rat skeletal muscle creatine kinase.

A series of cDNA clones corresponding to 1494 bases of rat muscle creatine kinase mRNA has been isolated and characterized. The identity of these clones has been confirmed by DNA sequence analysis and by comparison of the predicted amino acid sequence with that determined for the purified protein. The cDNA sequence accounts for the entire coding sequence of the creatine kinase protein in addition to the complete 3' untranslated region and 68 bases of 5' noncoding region. Sequences corresponding to the active site region of the protein, the initiation codon, the termination codon, and poly(A) addition signal have been identified.

Amino Acid Sequence↗

Sister-chromatid exchange and micronucleus induction as indicators of genetic damage in maternal and foetal cells.

The effectiveness of 3 compounds, procarbazine, mitomycin C and cyclophosphamide as inducers of sister-chromatid exchanges (SCEs) in granulocyte-macrophage progenitor cells, in foetal liver and bone marrow from pregnant mice at day 17 of gestation were determined. Cyclophosphamide and procarbazine induced similar SCE frequencies in maternal and foetal cells. Mitomycin C was slightly less effective in foetal liver than in maternal bone marrow. In contrast to the results of SCE induction, cyclophosphamide produced more micronucleated polychromatic erythrocytes in foetal liver than in bone marrow. The SCE results for mitomycin C and procarbazine are compared with results obtained previously for micronuclei induction in 15-day pregnant animals.

Animals↗

The human lymphocyte micronucleus assay. Response of cord blood lymphocytes to gamma-irradiation and bleomycin.

The induction of micronuclei in human cord blood lymphocytes by treatment with gamma-irradiation and bleomycin has been measured. Culture durations which gave peak MN frequencies were determined. The lowest tested doses, 0.1 Gy irradiation and 1.25 micrograms/ml bleomycin, produced significant increases in the frequency of micronuclei. The spontaneous frequency of micronucleated lymphocytes in 28 cord blood samples ranged between 0.5 and 9.5 per thousand lymphocytes, with a modal value of 2.5. The method is evaluated for its potential usefulness in monitoring populations for chromosome breakage.

Bleomycin↗

Short-term tests for transplacentally active carcinogens. Induction of sister-chromatid exchanges in foetal brain, lung and blood-forming cells by procarbazine and cyclophosphamide.

The induction of sister-chromatid exchanges (SCEs) by cyclophosphamide (CP) and procarbazine (PC) in mouse granulocyte-macrophage precursor cells (GM cells) and erythroblasts from foetal liver, and cells from foetal brain and foetal lung has been measured. Agents were administered in vivo, and cells explanted into BrdUrd-containing medium for 2 cell cycles in vitro (using specific growth-promoting substances where necessary) to determine SCE frequency. Tissue and cell-type differences in responses were observed, and it is concluded that the in vivo/in vitro transplacental SCE technique is a useful indicator of genotoxic effects of agents which are potential transplacental carcinogens.

Animals↗

Short-term tests for transplacentally active carcinogens. A comparison of sister-chromatid exchange and the micronucleus test in mouse foetal liver erythroblasts.

The effectiveness of 6 chemicals (benzo[a]pyrene, (BaP), cyclophosphamide (CP), diethylnitrosamine (DEN), methyl methanesulphonate (MMS), mitomycin C (MC) and procarbazine (PC) ) as inducers of micronuclei in foetal liver and maternal bone marrow erythroblasts has been determined, and related to that of gamma-radiation. CP, DEN, MMS and PC were all more effective in the foetal liver. The induction of micronuclei and SCEs by each chemical in foetal erythroblasts after in vivo exposure was measured. When expressed as induction of sister-chromatid exchanges (SCEs) per erythroblast/induction of micronuclei per erythroblast (/microM/kg), the ratios obtained were MC 580, BaP 470, DEN 430, CP 258, MMS 140 and PC 13. The lowest doses detected as potentially genotoxic by each test in foetal liver erythroblasts are (with the exception of PC which is a relatively ineffective inducer of SCEs) similar. When isolated foetal livers were exposed in vitro, SCE dose responses to BaP, MC, MMS and PC could be directly related to those from in vivo exposure, indicating the role of the foetal liver in metabolic activation, but CP was considerably more cytotoxic. The transplacental micronucleus test, and in vivo/in vitro method for SCEs in foetal liver erythroblasts, provide sensitive, complementary assays for genotoxic effects of chemicals during prenatal life. Since foetal liver possesses greater metabolic potential than adult bone marrow, the transplacental tests respond to genotoxic agents not detected by bone-marrow systems.

Animals↗

Amino acid sequence of the catalytic subunit of aspartate transcarbamoylase from Escherichia coli.

We propose a primary structure for the catalytic subunit of aspartate transcarbamoylase (aspartate carbamoyltransferase; carbamoylphosphate: L-aspartate carbamoyltransferase, EC 2.1.3.2) from Escherichia coli based on amino acid sequences of fragments obtained by cyanogen bromide cleavage, by tryptic digestion of the succinylated polypeptide, and by chymotryptic and proteinase C digestion of the intact catalytic chain. The protein contains 310 amino acids and has a calculated molecular weight of 33,944. The negatively and positively charged residues are distributed uniformly, and there is no indication of charge clustering in the linear sequence.

Amino Acid Sequence↗

Biological, chemical, and immunological studies of Rauscher ecotropic and mink cell focus-forming viruses from JLS-V9 cells.

Two murine leukemia viruses were isolated from JLS-V9 cells which had been infected with Rauscher plasma virus. One virus was XC positive and failed to grow on mink or cat cells and thus was an ecotropic virus. The other virus formed cytopathic foci on mink cells, was XC negative, and fell into the mink cell focus-forming (MCF) viral interference group and was thus an MCF virus. The glycoproteins of the two viruses could be distinguished immunologically, by peptide mapping, and by size in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The MCF virus produced gp69, and the ecotropic virus produced gp71, explaining the origin of the heterogeneous glycoprotein (gp69 and gp71) of Rauscher leukemia virus. Amino-terminal sequences of gp69 and gp71 were determined. The MCF sequence was distinct from the ecotropic sequence, but retained partial homology to it. The data show that the glycoproteins are encoded by related yet distinct genes. The protein structural data support the proposal that MCF virus gp70 molecules have nonecotropic sequences at the amino terminus, with ecotropic sequences occurring at the 3' end of the gene. The Rauscher MCF virus glycoprotein lacks a glycosylation site found at position 12 of the ecotropic sequence.

Animals↗

Short-term tests for transplacentally active carcinogens: sensitivity of the transplacental micronueleus test to diethylnitrosamine.

DEN is an established animal carcinogen, and is effective transplacentally; but like other nitrosamines, its genotoxicity is difficult to assess in established shortterm cytogenetic tests. The mouse transplacental micronucleus test demonstrates the ability of DEN to cause chromosome breakage in utero. Positive results were obtained from the 15th day of gestation and a linear dose-response relationship is expressed when micronucleated polychromatic erythrocytes are measured in neonatal blood, after transplacental exposure late in gestation. 25mg/kg (the lowest dose tested) caused a significant increase in MNPCEs (p less than 0.05). A micronucleus test based on polychromatic erythrocytes in neonatal blood permits maximal exploitation of the metabolic capacity of the prenatal liver and is therefore sensitive to clastogenic chemicals not activated by adult bone marrow.

Animals↗

Phenotypic characteristics of cell lines requiring interleukin 3 for growth.

A number of cell lines have been derived from bone marrow cultures in the presence of WEHI-3 conditioned media (CM) that continue to require WEHI-3 CM for growth in vitro. Because the WEHI-3 cell line has been shown to constitutively produce a lymphokine (IL 3) that induces the expression of 20 alpha-hydroxysteroid dehydrogenase (20 alpha SDH) in cultures of splenic lymphocytes from athymic mice, we examined whether these cell lines were dependent upon IL 3 for growth. The results demonstrate that the factor required for growth of these cell lines copurifies with IL 3 activity on G-100, DEAE cellulose, and CM cellulose column chromatography as well as in preparative isoelectric focusing and on hydrophobic supports in reverse phase high pressure liquid chromatography. The biologic activity of peak fractions in each case was similar in both types of assays. These results strongly suggest that the WEHI-3 CM-dependent cell lines are dependent on IL 3 for growth in vitro. All the cell lines have readily detectable levels of 20 alpha SDH but have differing cell surface phenotypes. The C3HSFFV line is devoid of conventional lymphoid cell surface markers with the exception of Lyt-1, whereas the FDC-P1 expresses Thy-1, Ly-5, and H-11. Other cell lines have intermediate phenotypes.

20-Hydroxysteroid Dehydrogenases↗

Mammalian enzymes of trimethyllysine conversion to trimethylaminobutyrate.

The biosynthesis of carnitine proceeds from trimethyllysine (TML) by beta-hydroxylation by a liver or kidney mitochondrial enzyme, which requires oxygen, alpha-ketoglutarate, ferrous iron, and ascorbate. This dioxygenase is rapidly inactivated by preincubation with Fe2+, but not Fe3+. The evidence suggests that superoxide anion is involved in the hydroxylation. beta-Hydroxytrimethyllysine undergoes aldol cleavage to glycine and trimethylaminobutyraldehyde under the influence of serine hydroxymethyltransferase and possibly a specific aldolase. The next step, the aldehyde oxidation, is catalyzed by a specific NAD-dependent aldehyde dehydrogenase from liver cytosol. The product, trimethylaminobutyrate, is then hydroxylated by a cytosolic dioxygenase to carnitine. This enzyme, which has the same cofactor requirements as TML hydroxylase, is found in the liver of all species examined, but is absent from the kidney of some species.

Aldehyde Oxidoreductases↗

Procedures for the purification of interleukin 3 to homogeneity.

A procedure is described for the routine purification of IL 3 to homogeneity from WEHI-3-conditioned media. The techniques employed include ammonium sulfate fractionation, DEAE-cellulose, hydroxylapatite, and G-75 Sephadex column chromatography. The last step in purification involves chromatography on C18 hydrophobic supports in RP-HPLC systems, which results in the coelution of a protein peak and IL 3 activity. This purification sequence results in approximately a 1,000,000-fold purification from the initial starting material with yields of 5 to 10% of the initial activity. typically, 150 liters of conditioned media yields 2 to 10 micrograms of IL 3. The purified material was homogeneous by SDS-PAGE analysis and had an apparent m.w. of 28,000. Purified IL 3 had a specific activity of approximately 0.05 ng/unit of activity. Additional criteria used to establish the relationship of the 28,000-dalton protein to IL 3 include the ability of an antiserum against IL 3 to concomitantly immunoprecipitate the iodinated protein and to inhibit its biologic activity as well as the ability of the iodinated protein to bind specifically to cell lines known to require IL 3 for growth.

Animals↗