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R Sen

Publications and source records attributed to R Sen.

At least 109 records · Page 6Linked to original sources

The NF-kappa B-binding site mediates phorbol ester-inducible transcription in nonlymphoid cells.

The mouse immunoglobulin kappa light-chain enhancer can interact with at least three independent nuclear proteins. One of these proteins, NF-kappa B, is constitutively present only in nuclear extracts derived from B cells and plasma cells. A DNA-binding protein with the same sequence specificity (and therefore presumed to be NF-kappa B itself) can be induced in pre-B cells, T cells, and nonlymphoid cells by phorbol 12-acetate-13-myristate (PMA); however, it is not clear whether the induced factor can activate transcription in nonlymphoid cells as NF-kappa B does in B cells. In this paper we show that multimerization of a fragment of the mouse kappa enhancer that carried only the binding site for NF-kappa B behaved like a B-cell-specific regulatory element. Furthermore, this unit served to activate transcription in nonlymphoid cells after treatment with PMA (but not with cyclic AMP derivatives), and the kinetics of transcription activation correlated well with the kinetics of factor induction. Thus, the induced DNA-binding activity appeared to be functionally indistinguishable from that of NF-kappa B.

B-Lymphocytes↗

Leukemoid reaction in erythema nodosum leprosum in a leprosy patient.

A case of lepromatous leprosy with erythema nodosum leprosum (ENL) presenting as a myeloid leukemoid reaction is reported. Very high leucocyte count with immaturity of the cells in myeloid series was present in peripheral blood. High leucocyte alkaline phosphatase score, absence of hepatosplenomegaly and transient nature of leukemoid reaction differentiated it from chronic myeloid leukemia and acute myeloblastic leukemia. The possible mechanisms of leukemoid reaction in ENL are discussed.

Adult↗

In vitro transcription of immunoglobulin genes in a B-cell extract: effects of enhancer and promoter sequences.

Transfection experiments have led to the identification of three DNA sequences that are responsible for the tissue-specific expression of immunoglobulin genes. As a first step toward characterizing these regulatory phenomena at the biochemical level, we report the development of an in vitro transcription system from cells of the B lymphoid lineage. In these extracts, transcription of the MOPC41 kappa promoter is correctly initiated and dependent on the presence of an upstream sequence element located between -44 and -79 base pairs from the cap site. Second, although standard in vitro transcriptions are not affected by the presence or absence of enhancer sequences, we observed that the addition of polyethylene glycol led to a B-cell extract-specific suppression of transcription from a template that carries an immunoglobulin enhancer.

B-Lymphocytes↗

Inducibility of kappa immunoglobulin enhancer-binding protein Nf-kappa B by a posttranslational mechanism.

NF-kappa B is a nuclear protein, found only in cells that transcribe immunoglobulin light chain genes, that interacts with a defined site in the kappa immunoglobulin enhancer. This protein can be induced in pre-B cells by stimulation with bacterial lipopolysaccharide (LPS). The induction involves a posttranslational activation, and the combined action of LPS and cycloheximide causes a superinduction. An active phorbol ester also induces this factor, and with kinetics more rapid than those for LPS stimulation. Phorbol-ester-mediated induction of NF-kappa B was observed in a T cell line (Jurkat) and a nonlymphoid cell line (HeLa), and is therefore not restricted to B-lymphoid cells. We interpret these results to indicate that factors that control transcription of specific genes in specific cells may be activated by posttranslational modification of precursor factors present more widely.

Anisomycin↗

Multiple nuclear factors interact with the immunoglobulin enhancer sequences.

To characterize proteins that bind to the immunoglobulin (Ig) heavy chain and the kappa light chain enhancers, an electrophoretic mobility shift assay with end-labeled DNA fragments was used. Three binding proteins have been found. One is NF-A, a factor found in all tested cell types that binds to the octamer sequence found upstream of all Ig variable region gene segments and to the same octamer in the heavy chain enhancer. The second, also ubiquitous, protein binds to a sequence in both the heavy chain and the kappa enhancers that was previously shown to be protected from methylation in vivo. Other closely related sites do not compete for this binding, implying a restriction enzyme-like binding specificity. The third protein binds to a sequence in the kappa enhancer (and to an identical sequence in the SV40 enhancer) and is restricted in its occurrence to B cells.

Animals↗

Characterization of mitochondrial monoamine oxidase of Ascaridia galli.

Oxidative deamination of various biogenic monoamines by Ascaridia galli monoamine oxidase (MAO) was blocked by different mammalian MAO inhibitors, namely, iproniazid, trans-PcP, nialamide and pargyline and the blockade was observed to be time as well as concentration dependent. The binding of inhibitors with chick ascarid MAO was of the irreversible type and the nature of the inhibition was competitive. Pargyline showed lowest I50 (8 microM) and Ki (12 microM) values. Chlorgyline and deprenyl at 100 microM concentration inhibited MAO by about 60 and 40% respectively, indicating the presence of both type A and type B MAO in A. galli.

Animals↗

Ascaris lumbricoides and Ascaridia galli: biogenic amines in adults and developmental stages.

Ascaris lumbricoides var. hominis and Ascaridia galli contain 5-hydroxytryptamine, histamine, dopamine, and norepinephrine. The chick parasite showed lower levels of monoamines compared to human ascaris. Amine concentrations in females were higher than in males. In all specimens, 5-hydroxytryptamine was the highest while norepinephrine was found to be uniformly low. The female reproductive organ contained the maximum amount of dopamine while intestine was rich in histamine. A progressive increase in the concentrations of biogenic amines was noticed during development.

Animals↗

Monoamine oxidase in adult Ascaridia galli.

Monoamine oxidase (MAO), catalysing oxidative deamination of biogenic monoamines, has been detected in adult Ascaridia galli. MAO was present in mitochondria and deaminated noradrenaline at the maximal rate, although serotonin, adrenaline, tyramine and dopamine were also degraded but more slowly. Of the organs studied, the body wall, female reproductive organ and intestine, the body wall (containing neuronal structures) showed highest MAO activity. Km value for chick ascarid mitochondrial MAO using tyramine as substrate was 1.66 X 10(-3) M and it was most active at 2.5 mM tyramine concentration, pH 7.5 and 40 degrees C. MAO of A. galli appeared to be thermolabile as nearly 80% of its activity was lost when the incubation temperature was increased 5 degrees above optimum.

Animals↗

Molecular mobilities and the lowered osmolality of the chromaffin granule aqueous phase.

Carbon-13 spin-lattice relaxation times, T1, have been measured in whole adrenal medullary tissue slices, in suspensions of isolated chromaffin granules, in the reconcentrated chromaffin granule lysate, and in various model solutions containing catecholamines. ATP, chromogranins and Ca2+. Reorientational correlation times have been calculated at 10 degrees C using T1 data and nuclear Overhauser enhancements for protonated carbons on both catecholamines and nucleotides. Correlation times in all media are relatively short and characteristic of highly fluid aqueous phases. Adrenalin and ATP exhibit substantial differences in correlation times in all media, however, the ratio tau R (ATP): tau R(catecholamine) ranging from 2.4 in simple 3:1 adrenalin-ATP solutions to 4 in intact chromaffin granules. This difference, as well as the relatively high absolute reorientational mobilities of both components, confirms the importance of labile ionic interactions between ATP and catecholamines, but rules out the presence of high concentrations of base-stacked structures. Participation of the chromogranins in ternary complexes with catecholamines and ATP appears to be of minor importance. Ionic interactions to the protein are not reflected in either 13C T1 values or chemical shifts of arginine or glutamate sidechain resonances, or in the 13C chemical shifts of ATP or catecholamines. Very labile protein-ATP binding appears to be reflected in the correlation time measurements, however, which show selective immobilization of ATP relative to catecholamine in the presence of soluble protein. Osmotic measurements indicate that solutions containing adrenaline, ATP and Ca2+ are highly nonideal, but probably not sufficiently so to account fully for the osmotic stabilization of the chromaffin through their polyelectrolyte properties, exert a significant influence on the intragranular osmolality. The osmotic lowering due to polyion-counterion interactions has been estimated semiquantitatively using a theory developed by Oosawa.

Adenosine Triphosphate↗

Water permeability of the chromaffin granule membrane.

NMR spin-lattice relaxation rates of solvent protons have been used to measure the water permeability coefficient of the chromaffin granule membrane. The technique involves labeling the chromaffin granule interior with Mn+2, which provides an efficient relaxation pathway for intravesicular solvent protons. Added Mn+2 spontaneously accumulates in the chromaffin granule matrix in the presence of the divalent cation-specific ionophore A23187 and is maintained against a large concentration gradient. In this way, the internal proton relaxation rate is readily augmented to values some 10(2)-10(3) times greater than that in the extravesicular water space. Transmembranal water transport permits solvent protons in the extravesicular water space, in which most of the observed NMR signal orginates, to sample the highly relaxive environment of the chromaffin granule matrix. By this process, water permeation shortens the observed relaxation rate. The diffusive water permeability coefficient of the chromaffin granule membrane has been measured over the temperature range 0-38 degrees C. The permeability coefficient measured at 25 degrees C is comparable to a previously reported value for planar lipid bilayers composed of ox brain lipids and cholesterol (Pd approximately equal to 0.37-0.53 10(-3)) cm X s-1 at 25 degrees C) but is substantially less than values for the plasma membranes of erythrocytes and Chlorella. Hypothesized hydrophilic "pores," thought to provide parallel permeation pathways in the latter membranes, appear to be absent in chromaffin granule membranes. The water permeation rate exhibits Arrhenius temperature behavior and does not reflect a phase transition at 32 degrees-34 degrees C observed previously in ESR spin-label studies of chromaffin granule ghosts.

Adrenal Medulla↗

The soluble components of chromaffin granules. A carbon-13 NMR survey.

Carbon-13 NMR spectra of the reconcentrated chromaffin granule lysate have been obtained at 50 MHz and 62.9 MHz. The spectrum contains a number of assignable resonances in addition to those of the main soluble components (catecholamines, adenine nucleotides and chromogranin). Guanine and uridine nucleotides are present at levels of 0.13 and 0.08 mol/mol adenine nucleotides, respectively. Concentrations of cytidine nucleotides and NAD+ are below the detection limit (0.02 mol/mol adenine nucleotides). An unidentified low molecular weight species, thought to be an adenine-containing oligonucleotide, is also present. Ascorbic acid was observed at a concentration of 0.14 mol/mol adenine nucleotides, but both dopamine and dehydroascorbic acid were below the detection limit. Protein resonances agree well with the reported amino acid composition of chromogranin A, with the exception of tryptophan and glutamine which have not previously been measured. The concentrations of these residues are estimated to be 12 +/- 3 and 39 +/- 5 residues per 77 000 dalton unit of chromogranin A. Substantial intensity due to unsaturated fatty acid side-chains in solubilized lipid is seen in the olefinic carbon region and in the methylene region, suggesting the presence of lipoprotein. Unassigned carbohydrate resonances are also present, but are largely obscured by sucrose in the isolation medium.

Adenine Nucleotides↗