In vitro vegetative propagation of Chinese cabbage

Explants from cotyledons, axillary buds, inflorescence stems and flower buds of Brassica campestris ssp. pekinensis (Lour.) Olsson (Chinese cabbage, cv. Wongbok) were cultured on MS medium with growth regulators. Multiple shoots were obtained from cotyledons, axillary buds and flower buds but not from inflorescence stems. Propagation of shoots from cotyledons was more successful than from axillary buds and flower buds. The vegetative propagation rates varied amongst clones derived from cotyledons of the same cultivar and seed lot. The propagation rates of the cotyledon-derived material followed a normal distribution with an average propagation rate of 2.6 shoots per two weeks subculture when cultured on MS media plus 44.4 m benzyladenine (BA) and 14.8 m -indolebutyric acid (IBA). Shoots from three clones were cultured on MS medium with nine different concentrations of BA. The concentration of BA which promoted the highest rate of shoot propagation varied for the three clones and was in the range 44.4 to 177.6 m.     

Nutrient budgets in a dry heathland watershed in northeastern Spain

Dissolved nutrient inputs in bulk precipitation and outputs in streamwater were measured during 3 years of contrasting hydrological conditions in a 6.3-ha, grazed heathland watershed on schists in the Montseny mountains (NE Spain), drained by an intermittent stream. On average, 39% of the precipitation became streamflow. Bulk precipitation delivered positive net alkalinity (mean 0.22 keq/ha/yr), sulphate input was moderate (9.0 kg SO₄-S/ha/yr), and the mean input of inorganic N was not exceptionally high (6.6 kg/ha/yr). Ion concentrations were relatively low in streamwater; SO₄ ^2- was the dominant anion. Most concentrations in streamwater varied seasonally, with maxima in late summer or early autumn and minima in spring. This pattern probably resulted from increased availability of ions for leaching due to decomposition of organic matter and chemical weathering during the warm period. Nitrate concentrations were relatively high in winter and dropped sharply in early spring, probably because of biological uptake. Annual element outputs in streamwater varied between years and seemed to be controlled by both the amount of annual streamflow and its seasonal distribution. Annual inputs exceeded outputs for dissolved inorganic N. The watershed accumulated H⁺ and Ca²⁺, had net losses of Na⁺ and Mg²⁺, and was close to steady state for K⁺, SO₄ ^2-, Cl^- and alkalinity. The chloride budgets gave no evidence of substantial dry deposition in this system. The cationic denudation rate was negative (-0.14 keq/ha/yr) because Ca²⁺ retention was higher than net exports of Na⁺ and Mg²⁺ from silicate weathering. Low nutrient export and little production of alkalinity suggest that this watershed has a low buffering capacity.     

Unfolding domains of recombinant fusion alpha alpha-tropomyosin.

The thermal unfolding of the coiled-coil alpha-helix of recombinant alpha alpha-tropomyosin from rat striated muscle containing an additional 80-residue peptide of influenza virus NS1 protein at the N-terminus (fusion-tropomyosin) was studied with circular dichroism and fluorescence techniques. Fusion-tropomyosin unfolded in four cooperative transitions: (1) a pretransition starting at 35 degrees C involving the middle of the molecule; (2) a major transition at 46 degrees C involving no more than 36% of the helix from the C-terminus; (3) a major transition at 56 degrees C involving about 46% of the helix from the N-terminus; and (4) a transition from the nonhelical fusion domain at about 70 degrees C. Rabbit skeletal muscle tropomyosin, which lacks the fusion peptide but has the same tropomyosin sequence, does not exhibit the 56 degrees C or 70 degrees C transition. The very stable fusion unfolding domain of fusion-tropomyosin, which appears in electron micrographs as a globular structural domain at one end of the tropomyosin rod, acts as a cross-link to stabilize the adjacent N-terminal domain. The least stable middle of the molecule, when unfolded, acts as a boundary to allow the independent unfolding of the C-terminal domain at 46 degrees C from the stabilized N-terminal unfolding domain at 56 degrees C. Thus, strong localized interchain interactions in coiled-coil molecules can increase the stability of neighboring domains.     

Use of a novel plasmid to monitor the fate of a genetically engineered Pseudomonas putida strain

Plasmid pSI30 was constructed to increase the sensitivity of detection of a genetically engineered micro-organism (GEM) and its recombinant DNA in environmental samples. This broad host-range, mobilizable plasmid contained chlorocatechol (clc) degradative genes, antibiotic resistance genes (ampicillin and kanamycin) and a fragment of eukaryotic DNA. The clc genes encode enzymes that convert 3-chlorocatechol to maleylacetic acid permitting the host, Pseudomonas putida RC-4, to grow on 3-chlorobenzoate. This catabolic phenotype was exploited using enrichment procedures to detect RC-4(pSI30) cells, free-living in the water column or when irreversibly bound to surfaces. The eukaryotic DNA sequence provided a unique target allowing positive identification by DNA:DNA hybridization. Using the eukaryotic DNA sequence as a probe, no transfer of the plasmid to indigenous bacteria was detected. Persistence of RC-4(pSI30) and its ability to multiply upon addition of 3-chlorobenzoate were demonstrated 78 days after its addition to natural freshwater. In flow-through microcosms RC-4(pSI30), undetectable as free-living cells, was found by enrichment as irreversibly bound sessile forms. These experiments revealed the stability of pSI30 and its utility in a 'combination' detection system for tracking the survival of a GEM and its DNA in environmental samples.     

人α1/158Ⅴ型(α1c型)干扰素基因在大肠杆菌中的表达及其产物的初步纯化

采用DNA聚合酶链反应(PCR)技术,将本实验室从中国人胎肝细胞染色体DNA中发现和分离的IFN-α1/158V基因的原始克隆,改造成适于进行非融合蛋白原核表达的结构形式,并在大肠杆菌中获得高效表达。测得重组IFN-α1/158V的抗病毒活性为1.9×10~7单位/升菌液。随后又采用以单克隆抗体亲和层析为主的纯化流程对表达产物进行初步纯化,获得了在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带的纯化产物。     

Effects of order of presentation of exercise intensities and of sauna baths on perceived exertion during treadmill running

Thirteen male subjects performed a running test on the treadmill consisting of four standard exercise intensities [65%, 75%, 85%, 95% maximal O2 uptake (VO2 max)] presented in ascending, descending or random order. At the end of each exercise intensity, O2 consumption, heart rate (fc), venous blood lactate concentration [( Ia]b) and perceived exertion were assessed. This last variable was determined according to the Borg nonlinear CR-20 scale. The same variables were also determined during exercise at a standard intensity (65% or 95% VO2 max) performed before and after a Finnish sauna bath. Ratings of perceived exertion showed a good test-retest reliability (r = 0.77); they were the same when the exercise intensity was expressed in relative (%VO2 max) or absolute (speed) terms, and were independent of the order of presentation of the exercise. The latter had no effect on fc either but it did, however, influence [Ia]b, which was significantly higher in the descending, as compared to the ascending or random modes of presentation. The sauna bath increased fc at a given exercise intensity, but left perceived exertion and [Ia]b unchanged. It was concluded that at least under the present experimental conditions, fc and venous [Ia]b do not play a major role as determinants of perceived exertion.     

Effect of lowered muscle temperature on the physiological response to exercise in men

The effect of low muscle temperature on the response to dynamic exercise was studied in six healthy men who performed 42 min of exercise on a cycle ergometer at an intensity of 70% of their maximal O2 uptake. Experiments were performed under control conditions, i.e. from rest at room temperature, and following 45 min standing with legs immersed in a water bath at 12 degrees C. The water bath reduced quadriceps muscle temperature (at 3 cm depth) from 36.4 (SD 0.5) degrees C to 30.5 (SD 1.7) degrees C. Following cooling, exercise heart rate was initially lower, the mean difference ranged from 13 (SD 4) beats.min-1 after 6 min of exercise, to 4 (SD 2) beats.min-1 after 24 min of exercise. Steady-state oxygen uptake was consistently higher (0.2 l.min-1). However, no difference could be discerned in the kinetics of oxygen uptake at the onset of exercise. During exercise after cooling a significantly higher peak value was found for the blood lactate concentration compared to that under control conditions. The peak values were both reached after approximately 9 min of exercise. After 42 min of exercise the blood lactate concentrations did not differ significantly, indicating a faster rate of removal during exercise after cooling. We interpreted these observations as reflecting a relatively higher level of muscle hypoxia at the onset of exercise as a consequence of a cold-induced vasoconstriction. The elevated steady-state oxygen uptake may in part have been accounted for by the energetic costs of removal of the extra lactate released into the blood consequent upon initial tissue hypoxia.     

Variation of geosmin content in Anabaena cells and its relation to nitrogen utilization

The addition of the proper amount of ammonium to the culture medium containing nitrate as nitrogen source enhanced the growth rate of Anabaena viguieri. The amount of geosmin produced by these cells varied with the concentrations of ammonium added. A negative correlation between the amount of geosmin produced and of the growth rate of cells was revealed. This was also found in cells grown on various forms of nitrogen sources. Without supply of any nitrogen compound, this organism is capable of fixing gaseous nitrogen, and under these conditions the cells grew relatively slowly. However, they produced more geosmin (per unit protein mass) than cells grown in the presence of combined nitrogen. The isolation of heterocysts, in which nitrogen was fixed, showed that these cells produced higher amounts of geosmin than vegetative cells. The possible relation of nitrogen assimilation to the production of geosmin in the cells was discussed.