β-adrenoceptors in human tracheal smooth muscle: characteristics of binding and relaxation

Specific binding of [¹²⁵I]-(−)-cyanopindolol to human tracheal smooth muscle membranes was saturable, stereo-selective and of high affinity (K_d=5.3±0.9 pmol/l and R_T=78±7fmol/g tissue). The β₁-selective antagonists atenolol and LK 203-030 inhibited specific [¹²⁵I]-(−)-cyanopindolol binding according to a one binding site model with low affinity in nearly all subjects, pointing to a homogeneous β₂-adrenoceptor population. In one subject using LK 203-030 a small β-adrenoceptor subpopulation could be demonstrated. The beta-mimetics isoprenaline, fenoterol, salbutamol and terbutaline recognized high and low affinity agonist binding sites. Isoprenaline's pK_H- and pK_L- values for the high and low affinity sites were 8.0±0.2 and 5.9±0.3 respectively. In functional experiments isoprenaline relaxed tracheal smooth muscle strips having intrinsic tone with a pD₂-value of 6.63±0.19.     

Density fluctuations in populations (1982–1986) and biological observations ofPotamopyrgus Jenkinsi in two trophically differing lakes

The hydrobiid snailPotamopyrgus jenkinsi (E.A. Smith), characterized by parthenogenesis and ovovivipary, was quantitatively sampled monthly between June, 1982, and December, 1986, on sandy bottoms in the shallow zones of the meso-oligotrophic Lake Maarsseveen I and the eutrophic Lake Maarsseveen II. The snail demonstrated a very clumped distribution in both lakes. The mean numbers of juveniles and adults taken together fluctuated strongly. Organisms in Lake I showed relatively high densities (up to 25,000 per m²) in 1982, followed by a sudden drop to values approaching zero in December, 1982, with a subsequent rapid increase in densities, fluctuating between 2,000 and 200 per m². In Lake II, densities of snails fluctuated between 13,000 and 300 per m² with decreases in the spring of 1985 and 1986. The various types of decreases in the lakes are extensively discussed, but no explanation is presently available. The reduction in Lake I was of catastrophic proportions, but the speed of recovery of the population was remarkable.Floating was observed only in Lake I, and only during the occurrence of the highest densities on the sediment. Burrowing behaviour was very common, but strongly suppressed under an uninterrupted dark regime. A shift of temperature from 15 to 22°C had the same effect. A number of submerged macrophyte species from Lake I proved to attractP. jenkinsi in the absence of sandy substrate, though these plants were only covered by the snail during the period of the highest densities in 1982. Temperatures of 20°C or lower were well tolerated, unlike temperatures of 25 and 30°C. Growth was distinct at 10, 15 and 20°C. Keeled individuals were encountered in much higher numbers in Lake I than in Lake II.     

13C-NMR studies on the influence of pH and nitrogen source on polyol pool formation in Aspergillus nidulans

Abstract The composition of the polyol pools in Aspergillus nidulans mycelium during active growth on sucrose depends strongly on pH. At pH 2.5, only mannitol is present. A comparison between nitrate- and ammonium-grown cultures shows stimulation of the arabitol content with nitrate a former nitrogen source. When starved mycelium is incubated either with natural-abundance or ¹³C-enriched glucose, label appears rapidly in mannitol and arabitol, regardless of the nitrogen source or the pH used.     

Effect of scatter factor on motility of epithelial cells and fibroblasts

The scatter factor is a protein released by fibroblasts that causes dispersal of epithelial cell colonies and disruption of intercellular junctions, as well as an alteration of morphology with ruffling and rapid extension and movement of pseudopodia. To find out if the scatter factor has a direct effect on cell migration, the Boyden chamber assay was used to determine the effect of partially purified factor on the migration of cells through 8 microns pore size filters. The results showed that the mobility of Madin-Darby canine kidney (MDCK) cells was stimulated, and usually maximal at 100 ng per ml, which is equivalent to 100 to 200 units of activity in the standard assay based on the morphology and arrangement of cells. The migration was due to chemotaxis and chemokinesis. A keratinocyte cell line was also sensitive as were, to a lesser extent, BSCl monkey kidney cells. The motility of freshly isolated fibroblasts and fibroblast cell lines, however, was not significantly affected. The results confirm the cell specificity and paracrine role of the scatter factor and show that this fibroblast-derived molecule can directly stimulate the movement of epithelial cells.     

Generation of soluble lignin-derived compounds during degradation of barley straw in an artificial rumen reactor

Summary The supernatants of effluents from an artificial rumen reactor degrading barley straw have been shown to contain lignin-derived compounds by UV spectral characteristics and pyrolysis mass spectrometry (PYMS). Most of these compounds were shown to be released by the action of rumen microorganisms. The compounds were quantified by measuring absorbance at 280 nm using bamboo-milled wood lignin as a standard. The concentration of the compounds rose from 0.5 mg·ml^–1 at solid and liquid retention times (SRT and HRT) of 60 and 12 h, respectively, and a loading rate (LR) of 25 g total solids (TS)·l^–1 per day to 3.5 mg·ml^–1 at a SRT of 144 h, an HRT of 20 days and an LR of 15 g TS·1^–1 per day. The highest concentration was below the level known to be toxic to rumen microorganisms in vitro. No indications were found for anaerobic lignin degradation in the rumen reactor. Offprint requests to: H. J. M. Op den Camp     

The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco

The effects of subcellular localization on single-chain antibody (scFv) expression levels in transgenic tobacco was evaluated using an scFv construct of a model antibody possessing different targeting signals. For translocation into the secretory pathway a secretory signal sequence preceded the scFv gene (scFv-S). For cytosolic expression the scFv antibody gene lacked such a signal sequence (scFv-C). Also, both constructs were provided with the endoplasmic reticulum (ER) retention signal KDEL (scFv-SK and scFv-CK, respectively). The expression of the different scFv constructs in transgenic tobacco plants was controlled by a CaMV 35S promoter with double enhancer. The scFv-S and scFv-SK antibody genes reached expression levels of 0.01% and 1% of the total soluble protein, respectively. Surprisingly, scFv-CK transformants showed considerable expression of up to 0.2% whereas scFv-C transformants did not show any accumulation of the scFv antibody. The differences in protein expression levels could not be explained by the steady-state levels of the mRNAs. Transient expression assays with leaf protoplasts confirmed these expression levels observed in transgenic plants, although the expression level of the scFv-S construct was higher. Furthermore, these assays showed that both the secretory signal and the ER retention signal were recognized in the plant cells. The scFv-CK protein was located intracellularly, presumably in the cytosol. The increase in scFv protein stability in the presence of the KDEL retention signal is discussed.