Detection of Glycogen-Debranching System in Trophozoites of Entamoeba histolytica

ABSTRACT. Homogenates of trophozoites of Entamoeba histolytica were shown to bring about the total degradation of glycogen while purified phosphorylase of the same source alone yielded a limit dextrin as end product. An enzyme system capable of debranching the limit dextrin was obtained from the 40,000 g pellet by extraction in aqueous medium, purified by gel filtration on Fractogel TSK HW-55(F), and separated from phosphorylase by chromatography on Blue Sepharose CL-6B and aminobutyl Agarose. The glycogen-debranching system was purified 540-fold to a state of homogeneity by criterion of disc-gel electrophoresis. The purified enzyme was able to degrade glycogen-limit dextrin in the presence of phosphorylase and exhibited activities of both amylo-1,6-glucosidase (EC 3.2.1.33) and 4- α -glucanotransferase (EC 2.4.1.25). Although amylo-1,6-glucosidase released glucose from a glycogen-phosphorylase limit dextrin, transferase activity moved single glucose residues from the limit dextrin to 4-nitrophenyl- α -glucoside yielding successively 4-nitrophenyl- α -maltoside and 4-nitrophenyl- α -maltotrioside that could be detected by HPLC. Native glycogen-debranching system exhibited a relative molecular mass of M_r= 180,000 ± 10% by gel filtration and gel electrophoresis in both denaturing and nondenaturating conditions.     

Declining body size in an endangered large mammal

Body size affects almost every aspect of the biology of a species. According to the ‘resource rule’, decreasing resource availability (e.g. prey density) will lead to a reduction in body size or, alternatively, a decline in mass‐independent energy expenditure. In the present study, we provide a test of this hypothesis, assessing the effect of significantly decreasing prey density on endangered African wild dog (Lycaon pictus) body size and energy expenditure over a 20‐year period. As predicted from the ‘resource rule’, decreasing resource availability resulted in energetic re‐allocation: wild dogs' body size decreased significantly (both shorter and slimmer), whereas our fitness‐related measure of energy expenditure (i.e. litter size) remained constant over time. A phenotypic change of up to 17% within 20 years, as found in the present study, appears to be unprecedented in a nonharvested large mammal, thus advancing the emerging field of eco‐evolutionary dynamics. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 8–12.     

Mate guarding in relation to seasonal changes in the energy reserves of two freshwater amphipods (Gammarus fossarum and G. pulex)

1. We assessed sex‐specific seasonal changes in major energy storage compounds (triglycerides, glycogen) in Gammarus fossarum and Gammarus pulex collected from the field, with respect to their reproductive activity. 2. The dynamics of stored energy followed a seasonal pattern in both species and sexes. Moreover, over a 4‐year period, these changes were independent of the year in which they were investigated. Stored energy reached a peak in late winter, but was depleted in late summer and early autumn, coinciding with the reproductive periods. 3. Triglyceride (annual mean ± SD) accounted for 79.7 ± 11.9% of the total stored energy and was responsible for the seasonal pattern. In contrast, glycogen contributed a lesser percentage (20.3 ± 11.9%). Over the study period, the amount of stored energy ranged between 0.39 and 4.08 kJ g^?1 dry mass (triglyceride: 0.19–3.69 kJ g^?1 dry mass; glycogen: 0.14–0.80 kJ g^?1 dry mass). 4. In both species, the energy reserves of males were drastically depleted shortly before the cessation of precopulatory mate guarding in the field, thus offering a bioenergetic explanation for the reproductive period in these two widespread species.     

Ultrastructural study of metamorphosis in the freshwater bryozoan Plumatella fungosa (Bryozoa,Phylactolaemata)

Summary

At metamorphosis the attachment of the Plumatella larva to the substrate is effected by secretions from glandular cells in the apical plate, the leading pole during swimming. The larval mantle folds back and slides down towards the substrate. By ciliary activity an adhesive secretion is spread over the metamorphosing larva and the attachment area. Two polypides appear through the larval terminal opening. The mantle fold, together with gland cells, nerve cells, sensory cells, and muscle cells from the larva form a nutritive cell mass. Reduction of this nutritive cell mass is accomplished by autolysis and phagocytosis. An invaginated area of the nutritive cell mass is provided with a dense layer of microvilli, which seem to have an absorbtive function. The nutritive cell mass consisting of transitory larval tissues provides a significant source of nutrient for the developing polypide buds.     

Rhizospheric NO affects N uptake and metabolism in Scots pine (Pinus sylvestris L.) seedlings depending on soil N availability and N source

We investigated the interaction of rhizospheric nitric oxide (NO) concentration (i.e. low, ambient or high) and soil nitrogen (N) availability (i.e. low or high) with organic and inorganic N uptake by fine roots of Pinus sylvestris L. seedlings by ¹⁵N feeding experiments under controlled conditions. N metabolites in fine roots were analysed to link N uptake to N nutrition. NO affected N uptake depending on N source and soil N availability. The suppression of nitrate uptake in the presence of ammonium and glutamine was overruled by high NO. The effects of NO on N uptake with increasing N availability showed different patterns: (1) increasing N uptake regardless of NO concentration (i.e. ammonium); (2) increasing N uptake only with high NO concentration (i.e. nitrate and arginine); and (3) decreasing N uptake (i.e. glutamine). At low N availability and high NO nitrate accumulated in the roots indicating insufficient substrates for nitrate reduction or its storage in root vacuoles. Individual amino acid concentrations were negatively affected with increasing NO (i.e. asparagine and glutamine with low N availability, serine and proline with high N availability). In conclusion, this study provides first evidence that NO affects N uptake and metabolism in a conifer.     

Aqueous pathways dominate permeation of solutes across Pisum sativum seed coats and mediate solute transport via diffusion and bulk flow of water

The permeability of seed coats to solutes either of biological or anthropogenic origin plays a major role in germination, seedling growth and seed treatment by pesticides. An experimental set‐up was designed for investigating the mechanisms of seed coat permeation, which allows steady‐state experiments with isolated seed coats of Pisum sativum. Permeances were measured for a set of organic model compounds with different physicochemical properties and sizes. The results show that narrow aqueous pathways dominate the diffusion of solutes across pea seed coats, as indicated by a correlation of permeances with the molecular sizes of the compounds instead of their lipophilicity. Further indicators for an aqueous pathway are small size selectivity and a small effect of temperature on permeation. The application of an osmotic water potential gradient across isolated seed coats leads to an increase in solute transfer, indicating that the aqueous pathways form a water‐filled continuum across the seed coat allowing the bulk flow of water. Thus, the uptake of organic solutes across pea testae has two components: (1) by diffusion and (2) by bulk water inflow, which, however, is relevant only during imbibition.     

Linking environmental warming to the fitness of the invasive clam Corbicula fluminea

Climate warming is discussed as a factor that can favour the success of invasive species. In the present study, we analysed potential fitness gains of moderate warming (3 °C above field temperature) on the invasive clam Corbicula fluminea during summer and winter. The experiments were conducted under seminatural conditions in a bypass-system of a large river (Rhine, Germany). We showed that warming in late summer results in a significant decrease in the clams' growth rates (body mass and shell length increase) and an increase in mortality rate. The addition of planktonic food dampens the negative effect of warming on the growth rates. This suggests that the reason for the negative growth effect of temperature increase in late summer is a negative energetic balance caused by an enhanced metabolic rate at limited food levels. Warming during early summer revealed contrasting effects with respect of body mass (no warming effect) and shell length (increased shell growth with warming). This differential control of both parameters further enhances the loss of the relative (size-specific) body mass with warming. In contrast, warming in winter had a consistently positive effect on the clams' growth rate as demonstrated in two independent experiments. Furthermore, the reproduction success (as measured by the average number of larvae per clam) during the main breeding period (April) was strongly enhanced by experimental warming during winter, i.e. by eight times during the relatively cold winter 2005/2006 and by 2.6 times during the relatively warm winter 2007/2008. This strong, positive effect of moderate winter warming on the clams' fitness is probably one reason for the recent invasion success of C. fluminea in the northern hemisphere. However, warm summer events might counteract the positive winter warming effect, which could balance out the fitness gains.