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T Johansen

Publications and source records attributed to T Johansen.

At least 109 records · Page 6Linked to original sources

Cloning and sequencing of the gene encoding the phosphatidylcholine-preferring phospholipase C of Bacillus cereus.

A synthetic oligodeoxynucleotide probe was used to clone the gene encoding the phosphatidylcholine-preferring phospholipase C of Bacillus cereus. The sequence of a 2050-bp restriction fragment containing the gene was determined. Analysis of the gene-derived amino acid (aa) sequence showed that this exoenzyme is probably synthesized as a 283-aa precursor with a 24-aa signal peptide and a 14-aa propeptide. The mature, secreted enzyme comprises 245 aa residues. Sonicates of Escherichia coli HB101 carrying the gene on a multicopy plasmid showed phospholipase C activity. This activity was inhibited by Tris, a known inhibitor of the B. cereus enzyme and also by antiserum raised against pure B. cereus phospholipase C. We conclude therefore that the gene is expressed in E. coli. The cloning and sequencing described here complete the first step toward using in vitro mutagenesis for investigations of the structure-function relationships of B. cereus phospholipase C.

Amino Acid Sequence↗

Nucleotide sequence of the Physarum polycephalum small subunit ribosomal RNA as inferred from the gene sequence: secondary structure and evolutionary implications.

The nucleotide sequence of the Physarum polycephalum small subunit ribosomal RNA (SSU rRNA) gene has been determined. Sequence data indicate that the mature 19S SSU rRNA is 1,964 nucleotides long. A complete secondary structure model for P. polycephalum SSU rRNA has been constructed on the basis of the Escherichia coli 16S rRNA model and data from comparative analyses of 28 different eukaryotic sequences. A "four-helix" model is presented for the central domain variable region. This model can be applied both to vertebrate and most lower eukaryotic SSU rRNAs. The increased size of P. polycephalum SSU rRNA relative to the smaller SSU rRNAs from such other lower eukaryotes, as Dictyostelium, Tetrahymena or Saccharomyces is due mainly to three G+C-rich insertions found in two regions known to be of variable length in eukaryotes. In a phylogenetic tree constructed from pairwise comparisons of eukaryotic SSU rRNA sequences, the acellular myxomycete P. polycephalum is seen to diverge before the appearance of the cellular myxomycete Dictyostelium discoideum.

Base Sequence↗

Alterations in sodium-potassium regulation in mononuclear leucocytes from young borderline hypertensive and offspring of hypertensive patients.

Membrane ion transports were investigated in lymphocytes from young normotensive and borderline hypertensive offspring with and without heredity for hypertension. Borderline hypertension per se was associated with an enhancement of sodium-potassium pump activity. Heredity per se was associated with increased sodium influx and ouabain-resistant sodium efflux.

Adolescent↗

86Rubidium uptake in mononuclear leucocytes from young subjects at increased risk of developing essential hypertension.

This study was designed to assess any changes in mononuclear leucocytes from young men at increased risk of developing essential hypertension and to determine whether any changes found were associated with borderline hypertension and/or heredity. To this end we used mononuclear leucocytes as a cellular model for in vitro measurement of total 86rubidium uptake to give an index of sodium-potassium pump activity. Four groups of subjects were evaluated, 28 normotensive and 20 borderline hypertensive offspring of hypertensives, and 12 borderline hypertensives and 28 normotensives with normotensive parents. 86Rubidium uptake was significantly increased in the borderline hypertensive subjects, especially in the borderline hypertensive offspring of hypertensive patients. Our results indicate that the sodium-potassium pump is activated in mononuclear leucocytes from borderline hypertensives, and especially in those borderline hypertensives with at least one hypertensive parent. The latter group was also the group at greatest risk of developing essential hypertension.

Adolescent↗

Energy metabolism in rat mast cells in relation to histamine secretion.

1. The relation between the energy metabolism and the secretory activity of rat peritoneal mast cells has been studied by determination of the cellular content of ATP and the rate of lactate production reflecting the rate of ATP synthesis under various experimental conditions. Secretion of histamine was induced by the antigen-antibody reaction, the polymeric amine compound 48/80, and the divalent ionophore A23187. 2. In presence of low concentrations of metabolic inhibitors (oligomycin or antimycin A) a linear relation between the secretion of histamine induced by all three liberators and the cellular ATP content at the time of cell activation was demonstrated. This may indicate a direct link between ATP and the secretory mechanism. 3. The possibility of an increased utilization of ATP during histamine secretion was explored in mast cells exposed to metabolic inhibitors. Incubation of mast cells with 2-deoxyglucose (2-DG) decreased the ATP content of the cells, and a long-lasting and stable level of mast cell ATP was observed. This is explained by a small decrease in the rate of ATP-synthesis by 2-DG. In 2-DG-treated cells secretion of histamine in response to compound 48/80 or the antigen-antibody reaction could still occur, and the secretion is shown to be associated with a decrease of the cellular ATP level. This ATP decrease indicates that secretion occurs by an ATP-requiring mechanism, that causes an increased cellular utilization of ATP in association with the secretory response. 4. The possibility has been considered that increased cytosol concentration of calcium in association with secretion may decrease the rate of ATP-synthesis due to accumulation of calcium by the mitochondria. This possibility can be excluded by the observation that in presence of respiratory inhibitors an identical ATP decrease was found during the secretion as in the case of the glycolytic inhibitor alone. This indicates that increased utilization of ATP by the cells was related directly to the secretory response. This observation was made with all three secretagogues. 5. A quantitative evaluation of the increased amount of ATP utilized by the cells in relation to the secretory response was performed by use of metabolic inhibitors. The ATP requirement of compound 48/80-induced secretion was 0.15 pmol/10(3) cells. This was observed when the ATP-synthesis was decreased to a negligible value.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Sodium homeostasis in lymphocytes and blood pressure alterations before and during salt restriction in normotensives and in essential hypertensives.

Blood pressure, lymphocytic sodium content and sodium efflux were studied in hypertensive and normotensive subjects during salt restriction. Diastolic blood pressure decreased significantly in both groups. In essential hypertension the initial high lymphocyte sodium content decreased during salt depletion with a simultaneous decrease in absolute sodium efflux probably due to primary changes in sodium influx. Conversely intralymphocytic sodium content increased in normotensive subjects, which may be caused by an inhibition of the sodium, potassium pump. Our findings indicate that autoregulatory mechanisms with regard to lymphocyte sodium metabolism differs between hypertensive and normotensive subjects.

Adult↗

Effect of oral salt loading on blood pressure and lymphocyte sodium metabolism in borderline hypertension.

A randomized double-blind cross-over trial was performed to test the effects of oral salt loading (normal diet + 200 mmol NaCl/day for 4 weeks followed by normal diet + 400 mmol/day for 1 week) against placebo on blood pressure and lymphocyte sodium homeostasis in 10 young borderline hypertensive men, genetically predisposed for essential hypertension. Salt loading caused no significant changes in blood pressure levels, lymphocyte sodium content and efflux. In conclusion, our subjects seem insensitive to a few weeks of excessive salt intake.

Blood Pressure↗

Effects of dimethylsulfoxide (DMSO), nocodazole, and taxol on mast cell histamine secretion.

Nocodazole depolymerized microtubules and increased the number of microfilaments, and dimethylsulfoxide increased the number of microfilaments. Both drugs inhibited compound 48/80-induced histamine release from rat mast cells. Taxol, which increased the number of microtubules, had no effect on histamine release. These observations support the view that microtubules may not be directly involved in secretion, but apparently an increased number of microfilaments is associated with a decreased capacity of the mast cells for histamine release. We suggest that microfilaments have to be depolymerized or rearranged before secretion can take place.

Alkaloids↗

Rubidium uptake of mononuclear leukocytes from normotensive and borderline hypertensive first degree relatives to patients with essential hypertension.

Uptake of 86Rubidium of mononuclear leukocytes (MNL) was used as a measure of cellular sodium-potassium pump activity. 86Rb-uptake was determined with the pump stimulated mainly from inside the cells by sodium as well as with a combined stimulation from inside by sodium and from outside by Rb. In the first case there was an increased pump activity in MNL from borderline hypertensive offspring of hypertensive patients (BHO), and this may be related to an increased number of pump sites observed previously (10). Estimation of maximal pump activity (Vmax) of MNL suggested that Vmax of each pump site in MNL from BHO may be decreased compared to control value of MNL from healthy normotensive subjects.

Humans↗

Measurement of specific [3H]-ouabain binding to different types of human leucocytes.

We have studied the specific binding of [3H]-ouabain to intact mononuclear leucocytes (82% lymphocytes) and polymorphonuclear leucocytes. In both types of cells [3H]-ouabain binding was saturable, confined to a single site of high affinity, slow to reach equilibrium, slow to reverse, temperature-dependent, competitively antagonized by potassium, and facilitated by the presence of divalent cations. The equilibrium dissociation constants were 2.4 +/- 0.7 nmol/l (polymorphs) and 2.4 +/- 0.4 nmol/l (mononuclear cells) (NS). The values of maximal specific ouabain binding, measured by Scatchard analysis of concentration vs binding curves (Bmax), were 33.9 +/- 6.0 fmol/10(6) cells (polymorphs) and 59.3 +/- 11.6 fmol/10(6) cells (mononuclear cells) (P less than 0.02). The corresponding numbers of sites per cell were 20415 +/- 3616 and 35712 +/- 6986 respectively (P less than 0.02). When the numbers of binding sites were expressed per square micron of cell surface area the difference between the two cell types was proportionately greater (83 and 186 sites per micron 2 respectively). We conclude that the [3H]-ouabain binding sites on mononuclear and polymorphonuclear leucocytes are similar in nature, but different in both number and density on the cell surface. Measurements of Bmax in mixed cell populations should therefore take account of cell type as well as cell size and number.

Binding, Competitive↗

Analgetic treatment in acute myocardial infarction. A controlled clinical comparison of morphine, nicomorphine and pethidine.

In a randomized double-blind study with flexible dosage, morphine, nicomorphine and pethidine were compared with regard to analgetic effect, dose requirements, dose intervals and adverse reactions. A total of 275 patients were included, and 28 patients were excluded due to adverse reactions (n = 16) and for practical reasons, etc. Acute myocardial infarction (AMI) was diagnosed in about 60% of the patients, and about 30% had ischemic heart disease without AMI. All three analgesics provided equally efficient pain relief in relative doses of morphine 10, nicomorphine 10 and pethidine 75 mg/ml. Severe adverse reactions were few (allergy 3 cases, respiratory insufficiency 4, severe bradycardia 4), whereas nausea was recorded in 20-30%, vomiting in 5-15% and dizziness in 10-30% of the patients, with no difference between the three drugs. Significant blood pressure drop (greater than 30 mmHg) was seen in 3-8% of the patients, with no significant differences between the drugs.

Aged↗

Ethacrynic acid inhibition of histamine release from rat mast cells: effect on cellular ATP levels and thiol groups.

The experiments concerned the effect of ethacrynic acid (0.5 mM) on the adenosine triphosphate (ATP) content of rat mast cells and the effect on histamine release induced by the ionophore A23187 (10 microM). Ethacrynic acid decreased the ATP level of the cells in presence of antimycin A and glucose as well as in presence of 2-deoxyglucose. A23187-induced histamine release was inhibited by ethacrynic acid, and this inhibition was completely reversed by dithiothreitol. These observations may indicate that ethacrynic acid inhibits glycolytic and respiratory energy production in rat mast cells. Furthermore, the inhibition of histamine release occurred mainly through interaction with thiol groups, although inhibition of cellular energy production may have been an additional mechanism.

Adenosine Triphosphate↗