Evidence that Frescon-induced contractures in Lymnaea stagnalis muscles do not depend on intracellular calcium stores: A comparison with caffeine action

The action of the molluscicide Frescon, which induces contractures in smooth and cross-striated muscles of the freshwater snail Lymnaea stagnalis, was compared with the action of caffeine. The results indicate that the latter agent promotes contractions in Lymnaea muscles by releasing Ca²⁺ from intracellular stores. Such a release could not be attributed to the action of Frescon. Furthermore, the molluscicide did not appear to alter muscle function at the level of the contractile apparatus as it did not affect the time course of caffeine-induced contractions. These results favor the hypothesis that Frescon increases sarcolemmal Ca²⁺ permeability.     

The acute toxicity of the death camas (Zigadenus species) alkaloid zygacine in mice, including the effect of methyllycaconitine coadministration on zygacine toxicity

Death camas (Zigadenus spp.) is a common poisonous plant on foothill rangelands in western North America. The steroidal alkaloid zygacine is believed to be the primary toxic component in death camas. Poisonings on rangelands generally occur in the spring when death camas is abundant, whereas other more desirable forage species are limited in availability. In most cases where livestock are poisoned by plants in a range setting, there is more than one potential poisonous plant in that area. One common poisonous plant that is often found growing simultaneously in the same area as death camas is low larkspur (Delphinium nuttallianum). Consequently, the objectives of this study were to conduct acute toxicity studies in mice and to determine if coadministration of low larkspur will exacerbate the toxicity of death camas. We first characterized the acute toxicity of zygacine in mice. The LD(50) of zygacine administered intravenously (i.v.) and orally was 2.0 ± 0.2 and 132 ± 21 mg/kg, respectively. The rate of elimination of zygacine from whole blood was determined to be 0.06 ± 0.01/min, which corresponds to an elimination half-life of 13.0 ± 2.7 min. The i.v. LD(50) of total alkaloid extracts from a Utah and a Nevada collection were 2.8 ± 0.8 and 2.2 ± 0.3 mg/kg, respectively. The i.v. LD(50) of methyllycaconitine (MLA), a major toxic alkaloid in low larkspur, was 4.6 ± 0.5 mg/kg, whereas the i.v. LD(50) of a 1:1 mixture of MLA and zygacine was 2.9 ± 0.7 mg/kg. The clinical signs in mice treated with this mixture were very similar to those of mice treated with zygacine alone, including the time of onset and death. These results suggest that there is an additive effect of coadministering these 2 alkaloids i.v. in mice. The results from this study increase knowledge and understanding regarding the acute toxicity of death camas. As combined intoxications are most likely common, this information will be useful in further developing management recommendations for ranchers and in designing additional experiments to study the toxicity of death camas to livestock.     

Determining phosphorus needs for cabbage grown on calcareous soils

Cabbage (Brassica olearacea var. capitata L.) yield differences from added P were highly significant up to the predicted response level of 0.4 mg 1 . The increases in yield were basically due to larger head sizes. No statistical differences were found after the soil P levels were adjusted above 0.4 ppm, as determined by P isotherms constructed from initial soil samples. Slight differences occurred between years, but likely were due to cultivar growth differences. Data show that the isotherm predicted level fits rather closely the P requirements for cabbage as related to crop yields. Since extraction methods measure only the intensity factor of P, these methods could lead to incorrect P fertilization recommendations. The absorption of P is progressively reduced as the temperature goes down.     

Protocol: High-throughput and quantitative assays of auxin and auxin precursors from minute tissue samples

ABSTRACT: BACKGROUND: The plant hormone auxin, indole-3-acetic acid (IAA), plays important roles in plant growth and development. The signaling response to IAA is largely dependent on the local concentration of IAA, and this concentration is regulated by multiple mechanisms in plants. Therefore, the precise quantification of local IAA concentration provides insights into the regulation of IAA and its biological roles. Meanwhile, pathways and genes involved in IAA biosynthesis are not fully understood, so it is necessary to analyze the production of IAA at the metabolite level for unbiased studies of IAA biosynthesis. RESULTS: We have developed high-throughput methods to quantify plant endogenous IAA and its biosynthetic precursors including indole, tryptophan, indole-3-pyruvic acid (IPyA), and indole-3-butyric acid (IBA). The protocol starts with homogenizing plant tissues with stable-labeled internal standards added, followed by analyte purification using solid phase extraction (SPE) tips and analyte derivatization. The derivatized analytes are finally analyzed by selected reaction monitoring on a gas chromatograph-mass spectrometer (GC-MS/MS) to determine the precise abundance of analytes. The amount of plant tissue required for the assay is small (typically 2--10 mg fresh weight), and the use of SPE tips is simple and convenient, which allows preparation of large sets of samples within reasonable time periods. CONCLUSIONS: The SPE tips and GC-MS/MS based method enables high-throughput and accurate quantification of IAA and its biosynthetic precursors from minute plant tissue samples. The protocol can be used for measurement of these endogenous compounds using isotope dilution, and it can also be applied to analyze IAA biosynthesis and biosynthetic pathways using stable isotope labeling. The method will potentially advance knowledge of the role and regulation of IAA.     

Neutral models for testing landscape hypotheses

Neutral landscape models were originally developed to test the hypothesis that human-induced fragmentation produces patterns distinctly different from those associated with random processes. Other uses for neutral models have become apparent, including the development and testing of landscape metrics to characterize landscape pattern. Although metric development proved to be significant, the focus on metrics obscured the need for iterative hypothesis testing fundamental to the advancement of the discipline. We present here an example of an alternative neutral model and hypothesis designed to relate the process of landscape change to observed landscape patterns. The methods and program, QRULE, are described and options for statistical testing outlined. The results show that human fragmentation of landscapes results in a non-random association of land-cover types that can be describe by simple statistical methods. Options for additional landscape studies are discussed and access to QRULE described in the hope that these methods will be employed to advance our understanding of the processes that affect the structure and function in human dominated landscapes.     

Comparison of the Sensitivity of Landscape-fire-succession Models to Variation in Terrain, Fuel Pattern, Climate and Weather

The purpose of this study was to compare the sensitivity of modelled area burned to environmental factors across a range of independently-developed landscape-fire-succession models. The sensitivity of area burned to variation in four factors, namely terrain (flat, undulating and mountainous), fuel pattern (finely and coarsely clumped), climate (observed, warmer & wetter, and warmer & drier) and weather (year-to-year variability) was determined for four existing landscape-fire-succession models (EMBYR, FIRESCAPE, LANDSUM and SEM-LAND) and a new model implemented in the LAMOS modelling shell (LAMOS(DS)). Sensitivity was measured as the variance in area burned explained by each of the four factors, and all of the interactions amongst them, in a standard generalised linear modelling analysis. Modelled area burned was most sensitive to climate and variation in weather, with four models sensitive to each of these factors and three models sensitive to their interaction. Models generally exhibited a trend of increasing area burned from observed, through warmer and wetter, to warmer and drier climates with a 23-fold increase in area burned, on average, from the observed to the warmer, drier climate. Area burned was sensitive to terrain for FIRESCAPE and fuel pattern for EMBYR. These results demonstrate that the models are generally more sensitive to variation in climate and weather as compared with terrain complexity and fuel pattern, although the sensitivity to these latter factors in a small number of models demonstrates the importance of representing key processes. The models that represented fire ignition and spread in a relatively complex fashion were more sensitive to changes in all four factors because they explicitly simulate the processes that link these factors to area burned. The US Government's and the Canadian Government's right to retain a non-exclusive, royalty-free license is acknowledged