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T Johansen

Publications and source records attributed to T Johansen.

At least 91 records · Page 5Linked to original sources
PubMed1990

Estimation of adenosine triphosphate utilization of rat mast cells during and after anaphylactic histamine secretion.

Determination of the cellular content of adenosine triphosphate (ATP) and the rate of ATP-synthesis were used to estimate the cellular utilization of ATP in relation to anaphylactic histamine secretion. There was an increased rate of oxidative ATP-synthesis and a decreased cellular ATP content during the time period of histamine secretion and immediately after its completion. During secretion the additional ATP-utilization above the basal level of ATP-synthesis was 0.51 pmol/10(3) cells. 2.5 min after cell activation, the rate of additional ATP-utilization was 0.30 pmol/10(3) cells/min, and the persistent ATP-decrease observed after 30 and 120 min may be due to a decreased rate of oxidative ATP-synthesis.

Adenosine Triphosphate↗
PubMed1990

Phospholipase C-mediated hydrolysis of phosphatidylcholine is an important step in PDGF-stimulated DNA synthesis.

Recent evidence suggests the involvement of phosphatidylcholine (PC) hydrolysis both in the control of normal cell growth and in transformation. We show here that the simple exogenous addition of Bacillus cereus PC-hydrolyzing phospholipase C (PC-PLC) is sufficient to elicit a potent mitogenic response in Swiss 3T3 fibroblasts by a mechanism that is independent of protein kinase C. Our results on the additivity and synergism between B. cereus PC-PLC, PDGF, and insulin in the mitogenic response indicate that this novel phospholipid degradative pathway may be important in the mitogenic signaling cascade activated by PDGF.

Animals↗
PubMed1990

Reversal by EGTA of the enhanced secretory responsiveness of mast cells due to treatment with ouabain.

The effect of EGTA on the enhancement by ouabain of compound 48/80-induced secretion from mast cells was compared with the effect on the Na(+)-K+ pump activity. The time-dependent secretory enhancement by ouabain was blocked by addition of EGTA to the cell suspension concomitantly with the addition of ouabain, and EGTA caused a large increase in the pump activity. Addition of 10 microM EGTA to ouabain-treated cells stopped but did not reverse the enhancement. The experiments show that the effect of ouabain was due to changes in a calcium pool utilized in compound 48/80-induced secretion following changes in the Na+,K+ pump activity.

Animals↗
PubMed1990

Organization of the mitochondrial genome of Atlantic cod, Gadus morhua.

The mitochondrial DNA (mtDNA) from the Atlantic cod, Gadus morhua, was mapped using 11 different restriction enzymes and cloned into plasmid vectors. Sequence data obtained from more than 10 kilobases of cod mtDNA show that the genome organization, genetic code, and the overall codon usage have been conserved throughout the evolution of vertebrates. Comparison of the derived amino acid sequences of proteins encoded by cod mtDNA to the ones encoded by Xenopus laevis mtDNA revealed that the amino acid identity range from 46% to 93% for the different proteins. ND4L is most divergent while COI is most conserved. GUG was found as the translation initiation codon of the COI gene, indicating a dual coding function for this codon. The sequences of the 997 base pair displacement-loop (D-loop)-containing region and the origin of L-strand replication (oriL), are presented. Only few of the primary and secondary structure features found to be conserved among mammalian mitochondrial D-loops, can be identified in cod. Presence of CSB-2 in the D-loop-containing region and the conserved hairpin structure at oriL, indicates that replication of bony fish mtDNA may follow the same general scheme as described for higher vertebrates.

Amino Acid Sequence↗
PubMed1990

Evidence for a role of phosphatidylcholine-hydrolysing phospholipase C in the regulation of protein kinase C by ras and src oncogenes.

The products of ras and src oncogenes are thought to be important components in pathways regulating cell proliferation and differentiation. In fibroblasts transformed by these oncogenes, increased diacylglycerol levels have been found which most probably arise from activation of the turnover of phosphatidylcholine. Diacylglycerol is a key activator of protein kinase C whose role in cell growth and transformation has been proposed. We demonstrate here by using immunochemical techniques that transformation by ras or src oncogenes is associated with permanent translocation of protein kinase C to the cytoplasmic membrane. However, no down-regulation of the enzyme is observed despite its permanent activation in these transformants. Importantly, the lack of down-regulation observed in ras and src transformed cell lines is mimicked by chronic treatment of NIH 3T3 fibroblasts with exogenous Bacillus cereus phosphatidylcholine-hydrolysing phospholipase C, but not with phorbol myristate acetate or exogenous Bacillus thuringiensis phosphatidylinositol-hydrolysing phospholipase C. These results strongly suggest that diacylglycerol derived from phosphatidylcholine but not from phosphoinositide turnover is responsible for the atypical regulation of protein kinase C in cell lines transformed by ras and src oncogenes.

Animals↗
PubMed1990

Beta-adrenoceptor responsiveness in intact leukocytes from adults and newborn infants; effect of phosphodiesterase inhibitors.

Intact mononuclear and polymorphonuclear leukocytes from adults and newborn infants were used to estimate the beta 2-adrenoceptor responsiveness in vitro. The accumulation of cAMP in response to receptor stimulation by isoprenaline was used as a measure of the receptor responsiveness. The relevance of the method is dependent on the activity of the cAMP phosphodiesterase inside the cells. The xanthine derivatives, theophylline (4 mmol l-1) and 3-isobutyl-l-methyl-xanthine (0.1-2.5 mmol l-1), were used to inhibit the cAMP phosphodiesterase activity. It appears that 3-isobutyl-l-methyl-xanthine at 2.5 mmol l-1 inhibits the phosphodiesterase activity almost completely. By use of this concentration no age-dependent changes in the isoprenaline-stimulated cAMP accumulation in mononuclear and polymorphonuclear leukocytes from adults and newborn infants could be demonstrated.

1-Methyl-3-isobutylxanthine↗
PubMed1990

Sodium content and sodium efflux of mononuclear leucocytes from young subjects at increased risk of developing essential hypertension.

Mononuclear leucocytes were used as a cellular model for the in vitro measurements of volume, sodium and potassium content, sodium efflux rate constants and absolute sodium efflux in order to assess any cellular changes in young men at increased risk of developing essential hypertension, and to analyze whether any such changes were associated with borderline hypertension and/or heredity. Four groups of subjects were evaluated: 28 normotensive (NTO) and 20 borderline hypertensive (BHO) offspring of hypertensives, 12 borderline hypertensives with normotensive parents (BH) and 28 normotensive subjects with normotensive parents (NT). The cellular sodium/potassium contents of the four groups were not discernibly different. Ouabain insensitive sodium efflux rate constant and corresponding absolute efflux were significantly increased in offspring of hypertensives. Ouabain sensitive absolute sodium efflux was significantly increased in borderline hypertensives (BHO + BH) compared to normotensives (NT + NTO). These results indicate that leucocytes from subjects predisposed to hypertension possess an increased ouabain insensitive sodium transport mechanism and in subjects with borderline hypertension the sodium-potassium pump seems activated.

Adolescent↗
PubMed1990

Ouabain inhibition of the sodium-potassium pump: estimation of ED50 in different types of human leucocytes in vitro.

1. We examined the effect of ouabain on the Na(+)-K+ pump of intact mononuclear leucocytes and polymorphonuclear leucocytes by using the radioactive potassium analogue, 86rubidium, as a tracer and measuring the cellular uptake of K+ (86Rb+). Na(+)-K+ pump activity was determined as the cellular uptake of K+ (86Rb+) that is sensitive to ouabain, 10(-5) mol l-1. 2. Dose-response curves for inhibition of the Na(+)-K+ pump were obtained after exposure of the cells to various concentrations of ouabain for 2.5 h when the level of pump inhibition was considered to be at steady state. 3. The ED50 of ouabain for the effect on the Na(+)-K+ pump was estimated to be 3 X 10(-9) mol l-1 in the absence of potassium. In the presence of potassium, 3 and 6.5 X 10(-3) mol l-1, it was increased by factors of 10 and 45 respectively. In the presence of potassium, 4 X 10(-3) mol l-1, the ED50 was similar but somewhat higher when determined in an artificial medium (Ringer solution) than when determined in autologous plasma. 4. The ED50 values observed in the present study were very similar to KD values reported in the literature on ligand binding of tritiated ouabain to the same types of human leucocytes.

Biological Transport, Active↗
PubMed1990

Inhibition of Na(+) -K+ pump activity by divalent cations in intact peritoneal mast cells of the rat.

1. The inhibition by the divalent cations magnesium, barium and strontium and the trivalent ion lanthanum of the Na(+) -K+ pump in the plasma membrane of rat peritoneal mast cells was studied in pure mast cell populations by measurement of the ouabain-sensitive uptake of the radioactive potassium analogue, 86rubidium (86Rb+). 2. Exposure of the cells to magnesium induced a time- and concentration-dependent decrease in the ouabain-sensitive K+(86Rb+)-uptake of the cells without influencing the ouabain-resistant uptake. The time-dependent decrease was apparent after incubation of the cells for 10 min or more, but no decrease was observed after 2 min incubation when the cells are supposed to be loaded with sodium due to the cell isolation procedure. 3. Barium and strontium caused concentration-dependent decreases in the ouabain-sensitive K(+) -(86Rb+) -uptake of the cells but the ouabain-resistant uptake was not changed. Half maximum decrease in the ouabain-sensitive K+(86Rb+)-uptake was observed with 1.8 mM magnesium, 1.2mM barium and 0.7 mM strontium. 4. The trivalent ion lanthanum blocked almost completely the ouabain-sensitive K+(86Rb+)-uptake at a concentration of 1 microM as does 1 mM calcium. Combining either of these ions with magnesium had no further inhibitory effect on the ouabain-sensitive uptake. 5. In conclusion, in addition to the previously suggested modulation by calcium of the activity of the Na+ (-)K+ pump, evidence is provided in this investigation that the modulation may be a more general effect of divalent and polyvalent cations present in the extracellular space through their influence on the sodium permeability of the plasma membrane.

Animals↗
PubMed1990

At least two types of control regions can be found among naturally occurring BK virus strains.

The various strains of BK virus (BKV) exhibit a remarkable degree of heterogeneity in the transcriptional control region, which may affect the biological characteristics of a BKV strain. We describe the detection and sequencing of BKV control regions directly from urine samples and after propagation in cell culture. A BKV strain [BKV (TU)] with a control region anatomy not described earlier, as well as a BKV (WW)-like strain [BKV (WWT)], was detected in urine samples by direct sequencing of polymerase chain reaction products. Urine inocula containing BKV (WWT) yielded BKV (TU) upon one passage in cell culture, while BKV (TU) did not change its control region during propagation in cell culture. Analysis of the nucleotide sequence of the transcriptional control regions revealed a partial deletion and duplication in BKV (TU) compared with BKV (WWT). In addition, the control region of BKV (TU) contains two point mutations relative to BKV (WWT). This indicates that both virus strains were probably present in the BKV (WWT)-dominated urine inocula, rather than that BKV (WWT) genomes were rearranged into BKV (TU) genomes during cell propagation. The heterogeneity of the control region of BKV strains is discussed in relation to both confirmed and putative transcription factor-binding sites.

Animals↗
PubMed1989

Members of the RTVL-H family of human endogenous retrovirus-like elements are expressed in placenta.

A cDNA clone homologous to the RTVL-H family of human retrovirus-like elements was isolated from a human placenta cDNA library. The nucleotide sequence of the 1084-bp cDNA revealed an open reading frame (ORF) that may encode a 146 amino acid protein with significant homology to retroviral proteases. Downstream from the putative protease ORF a 3' long terminal repeat (LTR) containing U3 and R regions was found. The cDNA sequence ends in a poly(A) tail appropriately positioned downstream from a polyadenylation signal in the LTR. Northern-blot analysis showed that several distinct RTVL-H homologous transcripts are present in human placenta. We also show that repetitive RTVL-H homologous sequences are present in the genomes of both gorilla and African green monkey.

Amino Acid Sequence↗
PubMed1989

Increased 22Na+-influx in lymphocytes from offspring of essential hypertensive patients.

Lymphocytes were used as a cellular model for the in vitro measurements of 22Na+-influx during sodium pump inhibition by ouabain. The measurements were made using lymphocytes from young men at increased risk of developing essential hypertension in order to assess any changes and to analyse whether any such changes were associated to borderline hypertension and/or heredity. Four groups were evaluated: 28 normotensive and 20 borderline hypertensive offspring of hypertensives, 12 borderline hypertensives and 28 normotensives with normotensive parents. 22Na+-influx was significantly increased in offspring of hypertensive parents especially in the normotensive offspring of hypertensive parents. The association between heredity and increased 22Na+-influx found by us in vitro may be caused by either an increased passive sodium-influx and/or an increased sodium-sodium exchange mechanism.

Adolescent↗
PubMed1989

Increased number of ouabain binding sites in lymphocytes from borderline hypertensives.

Lymphocytes were used as a cellular model for the in vitro measurements of maximal ouabain binding sites in order to assess any changes in young men at increased risk of developing essential hypertension, and to analyse whether any such changes were associated to borderline hypertension and/or heredity. Four groups were evaluated; 28 normotensive (NTO) and 20 borderline hypertensive (BHO) offspring of hypertensives. Twelve borderline hypertensives (BH) and 28 normotensive subjects (NT) with normotensive parents. The number of ouabain binding sites were significantly increased in the borderline hypertensives irrespective of heredity. The borderline hypertensives were heavier than the normotensives. A stepwise multiple regression model was therefore used in order to control confoundings by body mass index (BMI) and other factors such as age, gamma glutamyl transferase, 24 h sodium excretion, serum triglyceride, and serum cholesterol, which may influence the number of ouabain binding sites. Only BMI entered the stepwise model. These results indicate the presence of an increased number of sodium-potassium pumps in lymphocytes from borderline hypertensives. This difference may be attributed to the blood pressure disease or increased body mass.

Adolescent↗
PubMed1989

Na+ -K+ pump activity in rat peritoneal mast cells: inhibition by extracellular calcium.

1. Pure populations of rat peritoneal mast cells were used to study cellular potassium uptake. The radioactive potassium analogue, 86rubidium, was used as a tracer for potassium for measurements of the activity of the cellular potassium uptake process. 2. The ouabain-sensitive and the ouabain-resistant potassium (86rubidium) uptake of mast cells incubated in the presence of calcium, 1 mmol l-1, were very low, 52 and 147 pmol per 10(6) cells min-1. 3. Calcium-deprivation of the cells uncovered a large capacity ouabain-sensitive potassium (86rubidium) uptake mechanism. The activity of the uptake mechanism was decreased by reintroduction of calcium into the cell suspension, and it was dependent on cellular energy metabolism, temperature and pH. 4. The potassium (86rubidium) uptake of mast cells incubated in a calcium-free medium occurs through an active and ouabain-sensitive mechanism that has the nature of an enzyme, and it is mediated by the Na+ -K+ pump located in the plasma membrane. It is demonstrated that the activity of the Na+ -K+ pump mechanism is inhibited by low concentrations of extracellular calcium (0.1-1.2 mmol l-1). The possibility is discussed that calcium-deprivation may increase the pump activity by increasing the permeability of the plasma membrane for Na+.

Animals↗
PubMed1989

The mode of inhibition of the Na+-K+ pump activity in mast cells by calcium.

1 The inhibition by calcium of the Na(+)-K+ pump in the plasma membrane of rat peritoneal mast cells was studied in pure populations of the cells by measuring the ouabain-sensitive uptake of the radioactive potassium analogue, 86rubidium (86Rb+). 2 Exposure of the cells to calcium induced a time- and concentration-dependent decrease in the ouabain-sensitive K+(86Rb+)-uptake of the cells without influencing the ouabain-resistant uptake. The development of the inhibition required the presence of potassium in the medium in the millimolar range (1.5-8.0 mM), and it did not occur at a concentration of potassium (0.24 mM) that is probably rate limiting for the pump activity. In the presence of 1 mM calcium full inhibition developed almost immediately and was not readily reversed. The inhibition was not significantly reduced by 15 min incubation with 1.2 mM EGTA. 3 The inhibitory action of calcium did not develop when the mast cells were incubated in a potassium-free medium, which is known to block Na(+)-K+ pump activity and allow accumulation of sodium inside the cells. Likewise, increasing the sodium permeability of the plasma membrane by monensin abolished the inhibition of the pump activity. In both cases, incubation of the cells with 4.7 mM potassium and tracer amounts of 86Rb+ resulted in a very large uptake of K+ (86Rb+) into the cells (up to 2 nmol per 10(6) cells min-1), indicating a high activity of the Na(+)-K+ pump. 4. These observations support the view that long-term incubation of rat peritoneal mast cells in a calcium-free medium increases the permeability of the plasma membrane to sodium, and the consequent increase in the intracellular concentration of sodium causes an increase in the activity of the pump. Addition of calcium to the cell suspension decreases the sodium permeability, and hence the pump activity. This hypothesis is supported by the stimulation of pump activity produced by monensin, which is not inhibited by calcium. The enhancement of pump activity after exposure of calcium-deprived cells to EGTA might be the result of a further increase in the sodium permeability of the plasma membrane.

Animals↗
PubMed1989

Apparent phosphate retrieval system in Bacillus cereus.

Bacillus cereus secretes three different phospholipases C. We studied the effect of Pi levels in the growth medium on the production of these exoenzymes. Production of both phosphatidylcholine-preferring phospholipase C and sphingomyelinase C was repressed by Pi in the growth medium, whereas production of phosphatidylinositol phospholipase C was unaffected. We also found that B. cereus secretes a phosphate-repressed alkaline phosphatase activity. Together with a previously reported highly efficient, active uptake system for Pi, these three phosphate-repressed exoenzyme activities seem to be part of a phosphate retrieval mechanism that operates under growth-limiting concentrations of Pi. In natural soil systems, which are the natural habitats of B. cereus, the scarcity of Pi is the major growth-limiting factor. A phosphate-repressed metalloprotease activity was also detected in culture supernatants of B. cereus. It is unclear whether this exoenzyme activity also participates in the proposed phosphate-scavenging system.

Alkaline Phosphatase↗
PubMed1989

Reorganization of the subplasmalemmal cytoskeleton in association with exocytosis in rat mast cells.

The subplasmalemmal cytoskeleton in mast cells has been studied by scanning electron microscopy of the internal side of the plasma membrane. Rearrangement of the dense subplasmalemmal network of actin filaments took place following cell activation by compound 48/80 and secretion of histamine. The rearrangement was a withdrawal of the subplasmalemmal cytoskeleton from the exocytotic sites and the development of bare, filament-free areas around the sites. In calcium-depleted mast cells we demonstrated a dense network that was difficult to break. Activation of the calcium-depleted cells by compound 48/80 did not induce rearrangement of the network, and in parallel there was no secretion of histamine.

Animals↗