MALDI-TOF MS of phosphatidylethanolamines: different adducts cause different post source decay (PSD) fragment ion spectra

Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly applied to lipids. However, positional acyl chain analysis of lipids by MALDI was so far scarcely described. In this paper, the fragmentation behavior of phosphatidylethanolamine (PE) is investigated by using post-source decay (PSD) MS. In dependence on the investigated adduct, significant differences could be obtained. It will be shown that in particular the negative ion spectra enable the determination of the individual acyl chains as well as their positions (sn-1 or sn-2). Therefore, MALDI-TOF PSD spectra are a real alternative to more sophisticated MS/MS methods.     

Differential expression of the three multicopper oxidases from Myxococcus xanthus

Myxococcus xanthus is a soil bacterium that undergoes a unique life cycle among the prokaryotes upon starvation, which includes the formation of macroscopic structures, the fruiting bodies, and the differentiation of vegetative rods into coccoid myxospores. This peculiarity offers the opportunity to study the copper response in this bacterium in two different stages. In fact, M. xanthus vegetative rods exhibit 15-fold-greater resistance against copper than developing cells. However, cells pre-adapted to this metal reach the same levels of resistance during both stages. Analysis of the M. xanthus genome reveals that many of the genes involved in copper resistance are redundant, three of which encode proteins of the multicopper oxidase family (MCO). Each MCO gene exhibits a different expression profile in response to external copper addition. Promoters of cuoA and cuoB respond to Cu(II) ions during growth and development; however, they show a 10-fold-increased copper sensitivity during development. The promoter of cuoC shows copper-independent induction upon starvation, but it is copper up-regulated during growth. Phenotypic analyses of deletion mutants reveal that CuoB is involved in the primary copper-adaptive response; CuoA and CuoC are necessary for the maintenance of copper tolerance; and CuoC is required for normal development. These roles seem to be carried out through cuprous oxidase activity.     

Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L

Chagas disease is caused by Trypanosoma cruzi, being considered a public health problem. An alternative to combat this pathogen is the use of natural products isolated from fruits such as Eugenia uniflora, a plant used by traditional communities as food and medicine due to its antimicrobial and biological activities. Ethanolic extract from E. uniflora was used to evaluate in vitro anti-epimastigote and cytotoxic activity. This is the first record of anti-Trypanosoma activity of E. uniflora, demonstrating that a concentration presenting 50% of activity (EC(50)) was 62.76 μg/mL. Minimum inhibitory concentration (MIC) was ≤ 1024 μg/mL. Our results indicate that E. uniflora could be a source of plant-derived natural products with anti-epimastigote activity with low toxicity.     

Heat stress affects the distribution of JIM8-labelled arabinogalactan proteins in pistils of <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> cv Micro-Tom

Arabinogalactan-proteins are a family of highly glycosylated hydroxyproline-rich glycoproteins widely distributed in the plant kingdom and mainly located at the cell surface. Because of their great heterogeneity, abundance and ubiquitous localization, arabinogalactan-proteins are thought to play important and different roles in plant growth and development. Many evidences also indicate a role of arabinogalactan-proteins during reproduction as well as in response to stress conditions. In the present work, we investigated the distribution of arabinogalactan-proteins recognised by JIM8 antibody in pistils of Solanum lycopersicum cv Micro-Tom heat-stressed for 3 h at 42 °C at different developmental stages (5 and 10 days before anthesis). Our results indicate that high temperature strongly affects the distribution and content of arabinogalactan-proteins in stigma and ovule, particularly in samples heat-stressed 5 days before anthesis. In stigmas, cells exhibited an altered pattern of JIM8-labelled AGPs, dispersed and less abundant. In ovules, the embryo sac-surrounding cells showed a clear reduction in the content of JIM8-labelled arabinogalactan proteins. These evidences suggest that heat stress affects both content and distribution of AGPs. Considering the role of AGPs in plant reproduction, from the acquisition of stigmatic receptivity to pollen guidance into the ovule, we can suppose that high temperature affects all these processes through the alteration of AGPs.     

1-Aminocyclopropane-1-Carboxylate Oxidase Activity Limits Ethylene Biosynthesis in Rumex palustris during Submergence

Submergence strongly stimulates petiole elongation in Rumex palustris, and ethylene accumulation initiates and maintains this response in submerged tissues. cDNAs from R. palustris corresponding to a 1-aminocyclopropane-1-carboxylate (ACC) oxidase gene (RP-ACO1) were isolated from elongating petioles and used to study the expression of the corresponding gene. An increase in RP-ACO1 messenger was observed in the petioles and lamina of elongating leaves 2 h after the start of submergence. ACC oxidase enzyme activity was measured in homogenates of R. palustris shoots, and a relevant increase was observed within 12 h under water with a maximum after 24 h. We have shown previously that the ethylene production rate of submerged shoots does not increase significantly during the first 24 h of submergence (L.A.C.J. Voesenek, M. Banga, R. H. Thier, C.M. Mudde, F.M. Harren, G.W.M. Barendse, C.W.P.M. Blom [1993] Plant Physiol 103: 783-791), suggesting that under these conditions ACC oxidase activity is inhibited in vivo. We found evidence that this inhibition is caused by a reduction of oxygen levels. We hypothesize that an increased ACC oxidase enzyme concentration counterbalances the reduced enzyme activity caused by low oxygen concentration during submergence, thus sustaining ethylene production under these conditions. Therefore, ethylene biosynthesis seems to be limited at the level of ACC oxidase activity rather than by ACC synthase in R. palustris during submergence.     

Geographic control on phenotype expression. The case of Hybonoticeras mundulum (Oppel) from the Mexican Altiplano

Hybonoticeras mundulum (Oppel) (m) s.s. is reported from the Mexican Altiplano on the basis of material collected bed-by-bed in sections in the States of Durango and Zacatecas. The known range of this species in the Mexican Altiplano is interpreted to be uppermost Kimmeridgian (upper to uppermost Beckeri Zone) to lowermost Tithonian (basal to lower Hybonotum Zone). The Mexican specimens studied are the most complete and valuable collection of H. mundulum (Oppel) s.s. known from a given area, and reveal that Mexican populations show phenotypic features different from European ones. Vicariant events accord with data available about the areal and biostratigraphic distribution of this species, as well as with the combination of allocyclic and autocyclic factors influencing Mexican seas, the breaking of populations rather than colonization events, and the impoverished ammonite assemblages showing endemic traits within the stratigraphic interval studied.