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Biomedical subjects

A G Tielens

Publications and source records attributed to A G Tielens.

At least 19 recordsLinked to original sources

Fasciola hepatica miracidia are dependent on respiration and endogenous glycogen degradation for their energy generation.

It is generally accepted that free-living stages of parasitic helminths are dependent on aerobic degradation of endogenous energy sources for their energy generation. This concept, however, is not the result of extensive experimental evidence, but originated mainly intuitively as oxygen is widely available in their habitat and these stages generally have a small size. Schistosoma mansoni, the sole parasitic helminth whose energy metabolism has been studied throughout its life-cycle indeed has aerobically functioning free-living stages. However, large differences exist in energy metabolism between adult stages of distinct parasitic helminths, and caution should be taken in predicting that all free-living stages of all parasitic helminths have the same, aerobic energy metabolism. Hence, this report studied the energy metabolism of Fasciola hepatica miracidia and demonstrated that F. hepatica miracidia are also dependent on aerobic degradation of their endogenous glycogen stores by glycolysis and on Krebs cycle activity for energy generation. However, in contrast to S. mansoni, F. hepatica miracidia cannot function anaerobically, as inhibition of the respiratory chain blocked motility and carbohydrate degradation, and finally resulted in death of the miracidia. Therefore, this report demonstrated that differences exist between miracidia of distinct species, in pre-adaptation of their energy metabolism to the occasional hypoxic conditions within their next host.

Animals↗

R-O-C(triple bond)N species produced by ion irradiation of ice mixtures: comparison with astronomical observations.

We have investigated the effects induced by ion bombardment of mixtures containing nitrogen-bearing compounds at low temperatures. The results show the formation of a band at 2080 cm-1 in binary mixtures, NH3:CH4 and N2:CH4, which we attribute to HCN embedded in the organic residue formed by ion irradiation. In addition to this band, ternary mixtures containing an oxygen-bearing species (i.e., H2O) form a compound with a prominent absorption band at about 2165 cm-1 (4.62 microns). We ascribe this band to a nitrile compound containing O that is bonded to the organic residue. A detailed comparison of the laboratory results with astronomical data of the 4.62 microns absorption band in protostellar spectra shows good agreement in peak position and profile. Our experimental studies show that N2, which is a more likely interstellar ice component than NH3, can be the molecular progenitor of the carrier of the interstellar band. This is an alternative to the pathway by which UV photolysis of NH3-containing ices produces the 4.62 microns band and implies that ion bombardment may well play an important role in the evolution of interstellar ices. Here, we discuss the implications of our studies for the chemical route by which the carrier of the 4.62 microns band is formed in these laboratory experiments.

Ammonia↗

Titanium carbide nanocrystals in circumstellar environments.

Meteorites contain micrometer-sized graphite grains with embedded titanium carbide grains. Although isotopic analysis identifies asymptotic giant branch stars as the birth sites of these grains, there is no direct observational identification of these grains in astronomical sources. We report that infrared wavelength spectra of gas-phase titanium carbide nanocrystals derived in the laboratory show a prominent feature at a wavelength of 20.1 micrometers, which compares well to a similar feature in observed spectra of postasymptotic giant branch stars. It is concluded that titanium carbide forms during a short (approximately 100 years) phase of catastrophic mass loss (>0.001 solar masses per year) in dying, low-mass stars.

Astronomical Phenomena↗

Low-temperature crystallization of silicate dust in circumstellar disks.

Silicate dust in the interstellar medium is observed to be amorphous, yet silicate dust in comets and interplanetary dust particles is sometimes partially crystalline. The dust in disks that are thought to be forming planets around some young stars also appears to be partially crystalline. These observations suggest that as the dust goes from the precursor clouds to a planetary system, it must undergo some processing, but the nature and extent of this processing remain unknown. Here we report observations of highly crystalline silicate dust in the disks surrounding binary red-giant stars. The dust was created in amorphous form in the outer atmospheres of the red giants, and therefore must be processed in the disks to become crystalline. The temperatures in these disks are too low for the grains to anneal; therefore, some low-temperature process must be responsible. As the physical properties of the disks around young stars and red giants are similar, our results suggest that low-temperature crystallization of silicate grains also can occur in protoplanetary systems.

Cold Temperature↗

The interstellar 4.62 micron band.

We present new 4.5-5.1 micron (2210-1970 cm-1) spectra of embedded protostars, W33 A, AFGL 961 E, AFGL 2136, NGC 7538 IRS 9, and Mon R2 IRS 2, which contain a broad absorption feature located near 4.62 micron (2165 cm-1), commonly referred to in the literature as the "X-C triple bond N" band. The observed peak positions and widths of the interstellar band agree to within 2.5 cm-1 and 5 cm-1, respectively. The strengths of the interstellar 4.62 micrometers band and the ice absorption features in these spectra are not correlated, which suggests a diversity of environmental conditions for the ices we are observing. We explore several possible carriers of the interstellar band and review possible production pathways through far-ultraviolet photolysis (FUV), ion bombardment of interstellar ice analog mixtures, and acid-base reactions. Good fits to the interstellar spectra are obtained with an organic residue produced through ion bombardment of nitrogen-containing ices or with the OCN- ion produced either through acid-base reactions or FUV photolysis of NH3-containing ices.

Ammonia↗

Studies on phospholipid turnover argue against sloughing of tegumental membranes in adult Schistosoma mansoni.

The tegumental membrane complex of Schistosoma mansoni is the site of interaction between the parasite and the host. The tegument is involved in uptake of many nutrients, but also plays a crucial role in the evasion of the actions of the host immune system. Essential for the success of this evasion is maintaining the integrity of the tegumental membranes. The rate of turnover of phospholipids was investigated by pulse-labelling worms cultured in vitro, followed by additional incubation in the presence of unlabelled substrates. Tegumental membranes were isolated, characterized using antibodies against specific tegumental proteins, and analysed. It was demonstrated that the most prominent fatty acid found in tegumental phospholipids, palmitate, incorporated rapidly into the phospholipid fraction during a 30 min pulse labelling. In a subsequent 20 h chase with unlabelled substrates, the incorporated radioactivity was lost again from the tegumental membrane complex. This high turnover of palmitate was found to be limited to phosphatidylcholine (PC) only. The turnover was due to deacylation/reacylation, and not to the sloughing of membranes as is the case in schistosomula. It is speculated that this rapid turnover of PC in the tegument of adult schistosomes plays a new and important role in the immune evasion by the parasite.

Animals↗

Recognition of schistosome glycolipids by immunoglobulin E: possible role in immunity.

To investigate the role of antibody responses to (glyco)lipids in immunity to schistosome infection, lipids extracted from Schistosoma mansoni eggs and adult worms were fractionated, and the antibody isotype profile reactive to the fractionated lipids in a well-characterized S. haematobium-infected population was investigated. In tests of 10 plasma samples it was found that immunoglobulin G (IgG) reactivity was highest to the fraction containing ceramidepolyhexosides, whereas IgE reactivity was most prominent to both cerebroside- and ceramidepolyhexoside-containing fractions. The fraction containing ceramidepolyhexosides was then tested for reactivity with IgG subclasses and IgE in plasma samples from 66 S. haematobium-infected patients. Considering IgG4 and IgE, isotypes of particular interest in helminth infections, we found that both isotypes recognized egg (glyco)proteins in more than 90% of the infected subjects. However, in the case of glycolipids, IgE reactivity was much more prominent than IgG4 reactivity (found in 80 and 41% of the subjects, respectively). Furthermore, worm glycolipid-specific IgE prior to treatment of the subjects with praziquantel was negatively correlated with egg counts at 2 years posttreatment, indicating that IgE directed towards glycolipids could play an important role in resistance to reinfection.

Adolescent↗

Rapid separation and identification of phosphatidylethanolamine molecular species.

A novel high performance liquid chromatography method is presented for the separation and identification of intact molecular species of phosphatidylethanolamine (PE). After isocratic separation, detection of species can be achieved by measurement of UV absorbance as well as by the quantitative method of light scattering detection. A mathematical relationship exists between i) the relative retention time of a PE molecular species and ii) the number of carbon atoms and double bonds in the aliphatic groups of the species. This relationship can aid in the identification of the species. Furthermore, the absence of non-volatile components in the solvent allows the use of electrospray mass spectrometry to identify the eluting components and to establish the position of the individual radyl groups at the glycerol backbone. Using this method, samples of bovine heart PE (rich in plasmalogens) and rat liver PE (rich in diacyl species) have been analyzed.

Animals↗

Near-infrared spectroscopy of the proto-planetary nebula CRL 618 and the origin of the hydrocarbon dust component in the interstellar medium.

A new 2.8-3.8 micrometers spectrum of the carbon-rich protoplanetary nebula CRL 618 confirms the previous detection of a circumstellar 3.4 micrometers absorption feature in this object (Lequeux & Jourdain de Muizon). The high resolution and high signal-to-noise ratio of our spectrum allow us to derive the detailed profile of this absorption feature, which is very similar to that observed in the spectrum of the Galactic center and also resembles the strong 3.4 micrometers emission feature in some post-asymptotic giant branch stars. A weak 3.3 micrometers unidentified infrared band, marginally detected in the CRL 618 spectrum of Lequeux & Jourdain de Muizon, is present in our spectrum. The existence of the 3.4 micrometers feature implies the presence of relatively short-chained, aliphatic hydrocarbon materials (-CH2-/-CH3 approximately = 2-2.5) in the circumstellar environment around CRL 618. It also implies that the carriers of the interstellar 3.4 micrometers feature are produced at least in part in circumstellar material, and it calls into question whether any are produced by the processing of interstellar ices in dense interstellar clouds, as has been previously proposed. Other features in the spectrum are recombination lines of hydrogen, rotational and vibration-rotation lines of molecular hydrogen, and a broad absorption probably due to a blend of HCN and C2H2 bands.

Astronomical Phenomena↗

Ether lipids and their possible physiological function in adult Schistosoma mansoni.

Schistosomes have lost the capability to synthesize fatty acids de novo, but they can modify fatty acids by chain elongation. This has a profound effect on the molecular species composition of the two main phospholipid fractions of schistosomes, phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Molecular species of phospholipids are increasingly recognized as important mediators, or precursors thereof, in signal transduction, immune response modulation, and events like membrane fusion. As these are all important aspects of schistosome membranes and of the tegumental membranes in particular, we analysed the PE and PC molecular species of the tegumental membranes, the worm body and the blood of the host. With the aid of on-line mass spectrometry, we unequivocally identified a large number of PC and PE species in schistosomes, among which considerable amounts of plasmalogen species. This was unexpected, as this lipid subclass has been assumed to be absent in the parasite. Species, like (20:1-16:0) diacyl PC and (16:0-20:1) plasmalogen PE, found to be main constituents in schistosomes, were absent from the blood of the host. Large differences were also found between the molecular species composition of the tegumental membranes and the membranes of the worm body. In the tegumental membranes, 1-hexadecyl 2-palmitoyl PC was detected, which could possibly function as a precursor for platelet activating factor (PAF).

Animals↗

5-Octadecenoic acid: evidence for a novel type of fatty acid modification in schistosomes.

The lipid metabolism of schistosomes is characterized by several intriguing adaptations to a parasitic way of living. The surface of the parasite consists of two closely apposed phospholipid bilayers, a structure unique to blood flukes. Schistosomes do not synthesize fatty acids de novo, but are able to modify fatty acids, which they obtain from the host, by chain elongation. Here we present evidence that schistosomes are capable of another type of fatty acid modification, resulting in the formation of 5-octadecenoic acid [C18:1(5)]. This highly unusual fatty acid, which is absent in the blood of the host, was shown to be almost exclusively located in the outer membrane complex of the schistosome. Within these membranes, it was almost exclusively present in one molecular phospholipid species, 1-palmitoyl-2,5-octadecenoyl phosphatidylcholine [C16:0-18:1(5)PtdCho]. Apart from dipalmitoyl phosphatidylcholine, this was the most abundant phosphatidylcholine species in the outer membrane complex. The specific synthesis by the schistosome of C18:1(5) and the highly specific localization of this fatty acid to the tegumental membranes suggest an important tegument-mediated role for this lipid.

Animals↗

The electron transport chain in anaerobically functioning eukaryotes.

Many lower eukaryotes can survive anaerobic conditions via a fermentation pathway that involves the use of the reduction of endogenously produced fumarate as electron sink. This fumarate reduction is linked to electron transport in an especially adapted, anaerobically functioning electron-transport chain. An aerobic energy metabolism with Krebs cycle activity is accompanied by electron transfer from succinate to ubiquinone via complex II of the respiratory chain. On the other hand, in an anaerobic metabolism, where fumarate functions as terminal electron acceptor, electrons are transferred from rhodoquinone to fumarate, which is the reversed direction. Ubiquinone cannot replace rhodoquinone in the process of fumarate reduction in vivo, as ubiquinone can only accept electrons from complex II and cannot donate them to fumarate. Rhodoquinone, with its lower redox potential than ubiquinone, is capable of donating electrons to fumarate. Eukaryotic fumarate reductases were shown to interact with rhodoquinone (a benzoquinone), whereas most prokaryotic fumarate reductases interact with the naphtoquinones menaquinone and demethylmenaquinone. Fumarate reductase, the enzyme essential for the anaerobic functioning of many eukaryotes, is structurally very similar to succinate dehydrogenase, the Krebs cycle enzyme catalysing the reverse reaction. In prokaryotes these enzymes are differentially expressed depending on the external conditions. Evidence is now emerging that also in eukaryotes two different enzymes exist for succinate oxidation and fumarate reduction that are differentially expressed.

Anaerobiosis↗

Processing of icy mantles in protostellar envelopes.

We have obtained CO absorption profiles of several young stellar objects (YSOs), spanning a range of mass and luminosity, in order to investigate their ice mantle composition. We present the first detection of CO toward the class I YSO L1489 IRS in the Taurus dark cloud. In general, the CO profiles for YSOs show evidence for both processed and pristine ices in the same line of sight, strong indirect evidence for CO, is suggested in R CrA IRS 7, L1489 IRS, Elias 18, and GL 961E. Toward other sources (R CrA IRS 1, IRS 2, W33A, NGC 7538 IRS 9, Mon R2 IRS 2) CO is present in (nearly) pure form. We propose an evolutionary scenario to explain the chemical diversity of the icy mantles toward these objects.

Astronomical Phenomena↗

Trypanosomatidae produce acetate via a mitochondrial acetate:succinate CoA transferase.

Hydrogenosome-containing anaerobic protists, such as the trichomonads, produce large amounts of acetate by an acetate:succinate CoA transferase (ASCT)/succinyl CoA synthetase cycle. The notion that mitochondria and hydrogenosomes may have originated from the same alpha-proteobacterial endosymbiont has led us to look for the presence of a similar metabolic pathway in trypanosomatids because these are the earliest-branching mitochondriate eukaryotes and because they also are known to produce acetate. The mechanism of acetate production in these organisms, however, has remained unknown. Four different members of the trypanosomatid family: promastigotes of Leishmania mexicana mexicana, L. infantum and Phytomonas sp., and procyclics of Trypanosoma brucei were analyzed as well as the parasitic helminth Fasciola hepatica. They all use a mitochondrial ASCT for the production of acetate from acetyl CoA. The succinyl CoA that is produced during acetate formation by ASCT is recycled presumably to succinate by a mitochondrial succinyl CoA synthetase, concomitantly producing ATP from ADP. The ASCT of L. mexicana mexicana promastigotes was further characterized after partial purification of the enzyme. It has a high affinity for acetyl CoA (Km 0.26 mM) and a low affinity for succinate (Km 6.9 mM), which shows that significant acetate production can occur only when high mitochondrial succinate concentrations prevail. This study identifies a metabolic pathway common to mitochondria and hydrogenosomes, which strongly supports a common origin for these two organelles.

Acetates↗

Differences in energy metabolism between trypanosomatidae.

Although various members of the family Trypanosomatidae generate energy in a similar way, fundamental differences also exist and are not always recognized. In this review, Louis Tielens and Jaap Van Hellemond discuss the known differences in carbohydrate metabolism among trypanosomatids, and especially compare Leishmania with trypanosomatids such as Trypanosoma brucei and Phytomonas spp. Special attention will be paid to differences in end-products of carbohydrate degradation, to differences in anaerobic capacities between the various trypanosomatids and to the components of their respiratory chains, including the presence or absence of a plant-like alternative oxidase. Furthermore, evidence will be discussed which indicates that the succinate produced by trypanosomatids is formed mainly via an oxidative pathway and not via reduction of fumarate, a process known to occur in parasitic helminths.

Journal Article↗

Glucose Transport and Metabolism in Mammalian-stage Schistosomes.

Adult schistosomes transport nutrients from the host bloodstream across their outer body covering or tegument. The tegument is a cytologically unusual structure; it is a syncytium bounded externally by two lipid bilayer membranes. In this review, Patrick Skelly, Louis Tielens and Chuck Shoemaker reconsider our understanding of how glucose enters schistosomes across this unusual outer covering in the light of recent papers characterizing glucose transport proteins and glucose metabolism pathways in these parasites.

Journal Article↗

Quantitative analysis of phosphatidylcholine molecular species using HPLC and light scattering detection.

A number of HPLC chromatographic procedures can be used to separate intact molecular species of phosphatidylcholine (PC), but on-line quantification has remained problematic due to insensitivity of UV-detection for saturated species. Here, a new method is presented, separating all major PC molecular species from a variety of biological samples in intact form using a single, short and isocratic run. Species were separated on two RP18 reverse-phase columns in series and all species displayed an exponential relation between retention time and the percentage of acetonitrile or triethylamine in the mobile phase, allowing optimization of the mobile phase on a theoretical base, rather than on time-consuming test-runs. The use of triethylamine as a volatile additive instead of choline chloride allowed the use of light scattering detection. On a molar base, the response of the detector was invariant between species and allowed quantification of as little as 50 pmoles. The method was tested using phosphatidylcholines with widely different molecular species patterns, such a PC from rat liver, porcine pulmonary surfactant, bovine heart, boar sperm cells, and the parasite Schistosoma mansoni. As only volatile components are present in the solvents, individual molecular species can easily be recovered in pure form from the column effluent, enabling their further analysis (e.g., scintillation counting).

Acetonitriles↗

The incorporation, modification and turnover of fatty acids in adult Schistosoma mansoni.

Schistosoma mansoni is known to be unable to synthesize fatty acids and sterols de novo, but the parasite is capable of synthesizing phospholipids and triacylglycerols from precursors obtained from the host. The present study focuses on the dynamics of the incorporation of fatty acids in adult parasites. This study showed that fatty acids were rapidly metabolized into complex lipids and that oleate (18:1) was efficiently converted to eicosenoate (20:1) by chain elongation, whereas palmitate was not elongated at an appreciable rate. This chain elongation mainly involved fatty acids that were previously esterified to complex lipids. Furthermore it was shown that in adult parasites triacylglycerols do not serve as fatty-acyl donors in phospholipid synthesis as had been suggested to be the case in schistosomula, because: (1) Immediately after pulse-labelling the specific activity of fatty acids in phospholipids was higher than in triacylglycerols; and (2) the specific activity of eicosenoate, which had been formed by chain elongation of incorporated oleate. was higher in phospholipids than in triacylglycerols. Fatty acids that were esterified to phospholipids had a high turnover, in contrast to fatty acids esterified to triacylglycerols, which persisted for extended periods of time in this lipid class (days rather than hours).

Acylation↗