In Vitro Culture of Phytomonas Sp. Isolated from Euphorbia characias. Metabolic Studies by 1H NMR

ABSTRACT. We describe the in vitro culture of Phytomonas species isolated from Euphorbia characias . The best choice between tested media was SDM-79, in which promastigotes, after 6 days of culture, reached cell densities as high as 4 × 10⁷ cells/ml. Cells growing in LIT or MTL medium showed longer division times and lower cell densities. We succeeded in obtaining Phytomonas sp. amastigote and spheromastigote forms in modified GRACE's medium, yielding transformation rates of up to 70%. Electron microscopy studies were performed in order to characterize the ultrastructural features of these forms obtained in vitro. On the other hand, metabolic studies based on qualitative (nuclear magnetic resonance spectroscopy) and quantitative metabolic methods (enzymatic assays) showed that promastigote forms secreted mainly ethanol, acetate, glycine, glycerol, piruvate and succinate in SDM-79 medium, whereas the major metabolites found after transformation in modified Grace's medium were ethanol, acetate, glycine, piruvate and smaller amounts of glycerol.     

The Different Behavior of Diphtheria Toxin, Modeccin and Ricin in HeLa Cells Infected with Trypanosoma cruzi

ABSTRACT. We have studied the action of diphtheria toxin, modeccin and ricin on HeLa cells infected by Trypanosoma cruzi . Parasitized HeLa cells were resistant to diphtheria toxin and modeccin, whereas non-parasitized cells from the same cultures and control cultures showed cytopathological alterations. Protein synthesis, assayed by the incorporation of labelled methionine, diminished in toxin-treated control cultures but remained unaltered in the infected ones, compared to synthesis by untreated infected cells. Ricin, on the other hand, is a toxin that enters the cytoplasm by endocytosis. It has greater cytopathological effects in parasitized cells than in non-parasitized ones from the same cultures or uninfected control cells. Protein synthesis was inhibited in infected cultures treated with ricin.     

IMMUNOHISTOCHEMICAL LOCALIZATION OF GABA IN CHAMELEON RETINA (CHAMALEO CHAMALEO)

We used a policlonal antiserum against GABA and demonstated GABA-immunoreactivity (GABA-IR) in several populations of amacrine cells in the inner nuclear layer (INL), and other cells in the inner plexiform layer (IPL) of the central and peripheral retina of the chameleon. Horizontal cells do not contain GABA-IR and the chameleon retina is therefore an exception among non-mammals. GABA-IR was not seen in cell bodies in the position of photoreceptor, bipolar and interplexiform cells suggesting that GABA is not involved in synaptic transmission in the outer plexiform layer of chameleon retina.     

Species-specific phytoplankton responses to nutrients and zooplankton manipulations in enclosure experiments

1. In situ enclosure experiments were performed in the mesotrophic Bermejales reservoir to evaluate the algal response to changes in the nutrient supply and in the zooplankton size structure and density in a 2 × 2 factorial design. The experiments were conducted during the spring bloom of nanoplanktonic diatoms in 1989. 2. Nutrient enrichment promoted a great increase of phytoplankton biomass indicating a strong nutrient limitation on phytoplankton growth. Total phytoplankton biomass was significantly lower in the Daphina-added enclosures at a given nutrient level and strong direct an indirect effect of zooplankton on phytoplankton community structure and nutrient availability were observed. 3. Most of the nanoplanktonic species were effectively grazed but species with protective coverings and large size colonies were favoured by grazers and small chlorococcales were unaffected probably because of their compensatory high growth rates. The decrease in total biomass imposed by grazers is attributable mainly to the decrease of Cyclotella ocellata, the most abundant species. This taxon suffers two net effects of zooplankton: direct grazing and the indirect decrease of Si availability caused by the growth of C. ocellata which was promoted by P excretion by zooplankton. Indirect effects of grazers on Si availability should, therefore, be taken into account in explaining phytoplankton succession and community structure. 4. In this experiment grazers affected considerably the nanoplanktonic community in Bermejales reservoir. The extent which they were affected, however, depended not only on the algal size as a determinant of edibility but also greatly on the specific nutrient requirements and taxonomic features of the algal species.     

Nickel and Cadmium-related Changes in Growth, Plasma Membrane Lipid Composition, ATPase Hydrolytic Activity and Proton-pumping of Rice (Oryza sativa L. cv. Bahia) Shoots

Six-day-old rice plants (Oryza sativa L. cv. Bahia) were grownin the presence of 0.5 mol m^–3 Ni or Cd for 5 or 10 d.Five days after treatment, some plants were transferred to amedium containing no heavy metal for 5 or 10 d. Plasmalemmavesicles from shoots of treated, transferred (recovery experiments)and control plants were isolated, ATPase activity and proton-pumpingwere assessed, and free sterols and phospholipids determined.The ATPase hydrolytic activity was increased by 37% and 34%in 5 and 10 d Cd-treated plants, respectively; and by 66% in5 and 10 d Ni-treated plants. However, neither the initial rateof H⁺ transport nor the proton-pumping rate at steady-statewere significantly affected by the treatments. The relativeproportion (%) of the plasmalemma sterols campesterol and ⁵-avenasteroldecreased while sitosterol increased during heavy metal treatment.The overall plasmalemma phospholipid fatty acyl chain lengthand degree of unsaturation were also reduced. In experimentswhere plants recovered from Cd and Ni treatment, differencesbetween treated and control plants were reverted, particularlyin 10 d Ni-recovered plants. The possible involvement of lipidsin the regulation of the plasmalemma ATPase as well as the relationshipbetween growth, ATPase and adaptations to stress conditionsare discussed. Key words: Cadmium, nickel, sterols, phospholipids, ATPase     

Energy storage during the transition from endogenous to exogenous feeding in Australian redclaw crayfish Cherax quadricarinatus (Von Martens, 1898)

Summary

The Australian redclaw crayfish Cherax quadricarinatus hatches at an advanced stage. Two post-hatching stages rely on yolk reserves before the organism becomes a juvenile. After a juvenile appears, the yolk is depleted and young crayfish might spend a few days among female pleopods before starting to feed independently. During this phase, the juvenile organism needs alternative energy sources. Offspring from three females were analyzed with histological and histochemical techniques, from hatching until reaching the 2-week juvenile stage, to trace digestive system development and internal organ lipid and glycogen accumulation. Neutral lipids are stored in specialized cells of the hepatopancreatic tissue until the juvenile stage is reached. An increasing rate of accumulation was found. Glycogen was stored in hepatopancreas and tail muscle cells until post-embryo II, and no increasing or decreasing rate was found in storage functions.     

Ancient DNA reveals traces of Iberian Neolithic and Bronze Age lineages in modern Iberian horses

Multiple geographical regions have been proposed for the domestication of Equus caballus . It has been suggested, based on zooarchaeological and genetic analyses that wild horses from the Iberian Peninsula were involved in the process, and the overrepresentation of mitochondrial D1 cluster in modern Iberian horses supports this suggestion. To test this hypothesis, we analysed mitochondrial DNA from 22 ancient Iberian horse remains belonging to the Neolithic, the Bronze Age and the Middle Ages, against previously published sequences. Only the medieval Iberian sequence appeared in the D1 group. Neolithic and Bronze Age sequences grouped in other clusters, one of which (Lusitano group C) is exclusively represented by modern horses of Iberian origin. Moreover, Bronze Age Iberian sequences displayed the lowest nucleotide diversity values when compared with modern horses, ancient wild horses and other ancient domesticates using nonparametric bootstrapping analyses. We conclude that the excessive clustering of Bronze Age horses in the Lusitano group C, the observed nucleotide diversity and the local continuity from wild Neolithic Iberian to modern Iberian horses, could be explained by the use of local wild mares during an early Iberian domestication or restocking event, whereas the D1 group probably was introduced into Iberia in later historical times.     

Biosynthetic pathways of the pheromone of the Egyptian armyworm Spodoptera littoralis

Abstract Most insect pheromones comprise multicomponent blends of geometric or optical isomers, and one major question is how insects produce species‐specific ratios of components for successful reproductive isolation. Key enzymes suggested to be involved in pheromone biosynthesis are acetyl‐coenzyme A carboxylase and fatty acyl synthetase, chain‐shortening enzymes, desaturases, elongases, reductases, oxidases, and alcohol acetyl transferases. The female pheromone composition of the Egyptian armyworm Spodoptera littoralis (Boisd.) is highly dependent on the origin of the strain. In this review, we present a summary of the different reported pheromone compositions of the moth, including from our recent studies on this subject, as well as the biosynthetic routes to the different components and the molecular approaches involved. In addition, the key role played in the proposed biosynthetic pathways by a number of important biosynthetic enzymes, such as chain shortening enzymes, desaturases and alcohol acetyl transferases, is outlined, as well as the latest developments on the inhibition of these enzymes.