Lipophosphoglycan Is Present in Distinctly Different Form in Different Entamoeba histolytica Strains and Absent in Entamoeba moshkovskii and Entamoeba invadens1

ABSTRACT. Lipophosphoglycan has recently been demonstrated on the cell surface of Entamoeba histolytica strain HM-1:IMSS. A monoclonal antibody against this molecule had failed to react with some other strains of E. histolytica, including the strain Rahman. To determine if a structurally distinct lipophosphoglycan existed in Rahman, [³H]galactose-labeled glycoconjugates were electrophoresed through sodium dodecyl sulfate polyacrylamide gel electrophoresis. The electrophoretic pattern in Rahman was very different compared to that obtained with strains HM-1:IMSS and 200:NIH. A number of experiments including sensitivity to mild acid, nitrous acid and phosphoinositol-specific phospholipase C suggest that the Rahman glycoconjugate is indeed a lipophosphogylcan-like molecule but distinctly different from that of HM-1:IMSS. Mild acid-treated glycoconjugates from Rahman and HM-1:IMSS revealed the presence of neutral trisaccharides and monosaccharides in Rahman but not in HM-1:IMSS. Human immune sera from amoebiasis patients and a polyclonal antibody against HM-1:IMSS liphophosphoglycan both recognized Rahman glycoconjugate. Thus, while lipophosphoglycan molecules from the two strains share common epitopes, they are clearly distinct from each other. Molecules bearing resemblance to lipophosphoglycan could not be detected in other Entamoeba species, namely Entamoeba invadens and Entamoeba moshkovskii.     

Acclimation is beneficial at extreme test temperatures in the Danube crested newt, Triturus dobrogicus (Caudata, Salamandridae)

Despite many studies demonstrating the effect of acclimation on behavioural or physiological traits, considerable debate still exists about the evolutionary significance of this phenomenon. One of the unresolved issues is whether acclimation to warmer temperature is beneficial at treatment or at more extreme test temperatures. To answer this question, we assessed the effect of thermal acclimation on preferred body temperatures ( T _ps), maximum swimming and running speed, and critical thermal maximum ( CT _max) in the Danube crested newt ( Triturus dobrogicus ). Adult newts were kept at 15 °C (control) and 25 °C (treatment) for 8 weeks prior to measurements. We measured T _ps in an aquatic thermal gradient over 24 h, maximum speeds in a linear racetrack at six temperatures (5–33 °C), and CT _max in a continuously heated water bath. T _ps were higher in newts kept at 15 °C than in those kept at 25 °C. The maximum swimming speed did not acclimate. The maximum running speed at 30–33 °C was substantially higher in newts kept at 25 °C than in those kept at 15 °C. CT _max increased with the treatment temperature. Hence, we conclude that the acclimation response to warm temperature is beneficial not at treatment but at more extreme temperatures in newts.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 627–636.     

Do males with higher mating success invest more in armaments? An across‐populations study in damselflies

1. Males with higher mating success would be expected to invest more in traits that facilitate mating, leading to steeper allometry of those traits with respect to body size. Across‐population studies following latitudinal variation in male mating success are an excellent study system to address this question. 2. Males of the damselfly Lestes sponsa were used to investigate whether the allometric patterns of the length and width of the anal appendages, used for grasping the female prior to mating, corresponded to male mating success. Across a large latitudinal gradient, it was hypothesised that there is a larger investment in the grasping apparatus, i.e. a steeper allometric slope, following higher mating success. 3. Behavioural observations in field enclosures showed the highest mating success at high latitude, while there were no significant differences in mating success between the central and low latitudes. Positive allometry was found for the length of the anal appendages in high‐latitude males, while central‐ and low‐latitude males showed no significant regressions of the traits on body size. 4. These results partially support the hypothesis, as high‐latitude, more successful males invested more in the length (but not the width) of the grasping apparatus than did central‐ and low‐latitude males. Therefore, higher mating success might be facilitated by larger investment in armaments. Intraspecific studies on allometric patterns of traits that participate in mating success might offer new insights into the role of those traits in the reproductive behaviour of a species.     

Reconstruction of fig wasp mating structure: how many mothers share a fig?

Abstract.  1. Fig wasps (Hymenoptera: Agaonidae) represent an important model system for studies of sex ratio evolution, mainly because they may adjust their sex ratios in response to the numbers of ovipositing females (foundresses) that enter a fig and their clutch size.
2. Until recently, it was assumed that all foundresses fail to re-emerge from the figs that they have entered to oviposit, but there is increasing evidence that such re-emergence may be routine. The common practice of counting the number of dead foundresses present in a fig in order to deduce the number of foundresses is therefore questionable in species where failure to re-emerge has not been confirmed.
3. In this study, the alternative approach of microsatellite markers was used to reconstruct the within-fig breeding structure of a pollinating fig wasp by genetic analysis of the offspring. Broods of Liporrhopalum tentacularis , a species where foundresses regularly re-emerge from figs, were collected from figs of Ficus montana in their natural habitat in Indonesia as well as from an experimental glasshouse population in Leeds (U.K.).
4. The estimated foundress densities in the glasshouse population were similar to those in the field and ranged from one to six foundresses per brood.
5. Nearly 40% of all broods were produced by a single foundress, indicating that mating in these broods occurs exclusively between full siblings. High levels of inbreeding are therefore common in this species.     

Green Fluorescent Protein (GFP) as a Reporter Gene for the Plant Pathogenic Oomycete Phytophthora ramorum

ABSTRACT. Transgenic Phytophthora ramorum strains that produce green fluorescent protein (GFP) constitutively were obtained after stable DNA integration using a polyethylene glycol and CaCl₂-based transformation protocol. Green fluorescent protein production was studied in developing colonies and in different propagules of the pathogen to evaluate its use in molecular and physiological studies. About 12% of the GFP transformants produced GFP to a level detectable by a confocal laser scanning microscope. Green fluorescent protein could be visualized in structures with vital protoplasm, such as hyphal tips and germinating cysts. In infection studies with Rhododendron , one of the GFP expressing strains showed aggressiveness equal to that of the corresponding non-labelled isolate. Thus, GFP could be used as a reporter gene in P. ramorum . Limitations of the technology are discussed.     

Periphyton as alternative food source for the filter-feeding cladoceran Daphnia magna

1.  Daphnia magna , a well-studied primary consumer, is mainly known as a filter feeder. In this study, we investigated the ability of D. magna to use periphyton as an alternative food source to phytoplankton. We examined the development of laboratory populations fed with different food sources ( Desmodesmus subspicatus and/or periphyton or neither) over a period of 42 days, and observed the behaviour of the daphnids.
2.  The addition of periphyton to phytoplankton food led to an increase of daphnid population biomass. When fed with periphyton as the only food source, a small but stable D. magna population developed.
3.  The behaviour of daphnids fed with both food sources revealed a preference for feeding on D. subspicatus . Only below a concentration of D. subspicatus of approximately 0.05 mg C L⁻¹ (0.4 × 10⁷ cells L⁻¹) did D. magna use periphyton as an alternative food source.
4.  Periphyton showed distinct reactions to grazing by D. magna . The thickness of the periphyton layer was reduced from about 4 to 1 mm and we observed a change in species composition due to grazing.
5.  The ability of D. magna to graze on periphyton could serve to stabilize its population density and reinforce its competitive advantage over other cladocerans. By switching between food sources, D. magna can act as a coupler between pelagic and benthic habitats and food webs.     

Expression of Glial and Vimentin Type Intermediate Filaments in Cultures Derived from Human Glial Material

Several cultures established from biopsies of apparently normal adult human glial material showed no cells positive for glial fibrillary acidic protein (GFA) when examined after seven or more cumulative population doublings (CPD), although the established glioma line U251 MG showed ∼3% GFA-positive cells, and U333 CG/343 MG clone 3 showed >98% GFA-positive cells. Both the human glia derived cultures and the glioma lines were positive when assayed with sera specific for vimentin. We therefore investigated the expression of GFA as a function of cumulative population doublings after the establishment of primary cultures. Under our experimental conditions, although GFA-positive cells were clearly present in the primary cultures accounting for some 3%–14% of the cells present, the GFA marker was subsequently lost, and the proliferating cultures expressed only the vimentin type of intermediate filament. Those cells that were GFA-positive also appeared to be vimentin-positive. GFA expression was not reinduced in cultures that had lost the GFA marker by treatment with dibutyryl cyclic AMP. We discuss two alternate hypotheses for the origin of the GFA-negative cells: (1) the cultures are of astrocyte origin but lose the ability to express GFA on culturing; (2) the cultures originate from cells of nonastrocyte origin present in the primary material.     

In comparison with nitrate nutrition,ammonium nutrition increases growth of the frostbite1 Arabidopsis mutant

Ammonium nutrition inhibits the growth of many plant species, including Arabidopsis thaliana. The toxicity of ammonium is associated with changes in the cellular redox state. The cellular oxidant/antioxidant balance is controlled by mitochondrial electron transport chain. In this study, we analysed the redox metabolism of frostbite1 (fro1) plants, which lack mitochondrial respiratory chain complex I. Surprisingly, the growth of fro1 plants increased under ammonium nutrition. Ammonium nutrition increased the reduction level of pyridine nucleotides in the leaves of wild‐type plants, but not in the leaves of fro1 mutant plants. The observed higher activities of type II NADH dehydrogenases and cytochrome c oxidase in the mitochondrial electron transport chain may improve the energy metabolism of fro1 plants grown on ammonium. Additionally, the observed changes in reactive oxygen species (ROS) metabolism in the apoplast may be important for determining the growth of fro1 under ammonium nutrition. Moreover, bioinformatic analyses showed that the gene expression changes in fro1 plants significantly overlap with the changes previously observed in plants with a modified apoplastic pH. Overall, the results suggest a pronounced connection between the mitochondrial redox system and the apoplastic pH and ROS levels, which may modify cell wall plasticity and influence growth.     

THE BIOSTIMULATORY EFFECT OF RED LASER IRRADIATION ON PIG BLOOD PLATELET FUNCTION

The molecular mechanisms of laser-induced changes in the cell structure and function are not well known. The authors examined the effects of low-power laser irradiation on unnucleated pig blood platelets. The obtained results showed that laser irradiation (1–5J) caused in blood platelets lipid peroxidation (measured as thiobarbituric acid reactive substances) and super-oxide anion generation, concomitant with the release of adenine nucleotides and proteins from platelets. The maximum platelet response to laser irradiation was observed when doses of 1.8–2J were used. Our results indicate that red laser irradiation induces: (1) platelet secretory process and the release of substances stored in the specific granules (adenine nucleotides, proteins); and (2) lipid peroxidation partly due to stimulation of endogenous arachidonate and production of its metabolites reacting with thiobarbituric acid.     

Campylobacter fetus Induced Proinflammatory Response in Bovine Endometrial Epithelial Cells

Campylobacter fetus subsp. fetus is the causal agent of sporadic abortion in bovines and infertility that produces economic losses in livestock. In many infectious diseases, the immune response has an important role in limiting the invasion and proliferation of bacterial pathogens. Innate immune sensing of microorganisms is mediated by pattern-recognition receptors (PRRs) that identify pathogen-associated molecular patterns (PAMPs) and induces the secretion of several proinflammatory cytokines, like IL-1β, TNF-α, and IL-8. In this study, the expression of IL-1β, TNF-α, IL-8, and IFN-γ in bovine endometrial epithelial cells infected with C. fetus and Salmonella Typhimurium (a bacterial invasion control) was analyzed. The results showed that expression levels of IL-1β and IL-8 were high at the beginning of the infection and decreased throughout the intracellular period. Unlike in this same assay, the expression levels of IFN-γ increased through time and reached the highest peak at 4 hours post infection. In cells infected with S. Typhimurium, the results showed that IL8 expression levels were highly induced by infection but not IFN-γ. In cells infected with S. Typhimurium or C. fetus subsp. fetus, the results showed that TNF-α expression did not show any change during infection. A cytoskeleton inhibition assay was performed to determine if cytokine expression was modified by C. fetus subsp. fetus intracellular invasion. IL-1β and IL-8 expression were downregulated when an intracellular invasion was avoided. The results obtained in this study suggest that bovine endometrial epithelial cells could recognize C. fetus subsp. fetus resulting in early proinflammatory response.