PubMed Health⌕ Search

Biomedical subjects

M Benchimol

Publications and source records attributed to M Benchimol.

At least 55 records · Page 3Linked to original sources

Morphogenesis of the hydrogenosome: an ultrastructural study.

The morphogenesis of hydrogenosomes in several trichomonad species (Tritrichomonas foetus, Trichomonas vaginalis, Tritrichomonas suis, Trichomonas gallinae, Tritrichomonas augusta and Monocercomonas sp) was investigated by transmission electron microscopy of thin sections and freeze-fracture replicas of whole cells or the isolated organelle. Close proximity, and even continuity, between endoplasmic reticulum and hydrogenosomes was observed. Structures were seen connecting hydrogenosomes to each other and to cytoplasmic structures. Morphological evidence is presented showing that in all the trichomonads here studied, hydrogenosomes, like mitochondria, may divide by two distinct processes: segmentation and partition. In the segmentation process, the hydrogenosome grows, becoming enlongated with the appearance of a constriction in the central portion. Microfibrillar structures appear to help the furrowing process, ending with a total fission of the organelle. In the partition process, the division begins by an invagination of the inner hydrogenosome membrane, forming a transversal septum, separating the organelle matrix into two compartments. We suggest that myelin-like structures seen either in close contact with or in the vicinity of the hydrogenosomes may be a source of membrane lipids for hydrogenosome growth.

Animals↗

Carbohydrate involvement in the association of a prokaryotic cell with Trichomonas vaginalis and Tritrichomonas foetus.

Previous studies have shown that Trichomonas vaginalis are capable of ingesting bacteria. This observation was confirmed in the present study and extended to Tritrichomonas foetus. Using a special strain of Escherichia coli grown under conditions that produced fimbriae presenting a lectin-like molecule recognizing mannose, we showed that the bacteria attached to and were ingested by trichomonads through a mechanism involving cell-to-cell recognition. Absence of the fimbriae or addition of alpha-methyl-D-mannoside to the interaction medium blocked attachment of the bacteria to the protozoa cell surface. Ingested bacteria were later digested within the cytoplasmic vacuole.

Animals↗

[Coma and transtentorial herniation syndrome due to acute non-expansive hemispheric lesion].

An 81-year-old woman developed left hemiplegia and coma due to a right hemisphere infarct and died 6 days later. When first seen in coma she had the classical signs of descending central herniation in the diencephalic stage. The CT scan of the third day showed a large hypodense area occupying the superficial and deep territories of the middle and anterior cerebral arteries on the left as well as a frontal cortico-subcortical hypodense area indicative of an older infarct on the opposite hemisphere. No mass effects were apparent. She followed a downhill course with signs of brainstem deterioration. A second CT scan a few hours before death revealed the expected pattern of brain shift and herniations. This case adds to the available evidence showing that the clinical signs of encroachment of supratentorial structures upon the basal forebrain can be reproduced by an acute mono-hemispheral lesion without visible mass effects. It indicates, further, that the role of intracranial displacements in the genesis and short-term prognosis of coma remains an unsettled matter. In at least a few number of cases, diaschisis might play a major role.

Acute Disease↗

Fine structure of Tritrichomonas foetus as seen using cryotechniques.

Tritrichomonas foetus was studied using different physical and chemical fixation methods such as fast-freezing (by high pressure, "slam-freezing," and jet-propane), freeze-substitution, conventional freeze-fracture and deep-etching, cryoultramicrotomy, and routine preparation for transmission electron microscopy. The use of fast-freezing fixation (FFF) proved to be superior in terms of structural preservation due to the rapidity of this fixation compared to that obtained using conventional chemical fixation. The low temperature techniques used here were useful to confirm data already obtained by conventional freeze-fracture using chemical fixation and cryoprotection, such as the presence of flagellar rosettes and costa structure. Cryoultramicrotomy and slam-freezing also demonstrated the presence of hair-like structures projecting out from the protozoan surface. New aspects of organelles of T. foetus were demonstrated.

Animals↗

Further studies on the endocytic activity of Tritrichomonas foetus.

The endocytic activity of Tritrichomonas foetus was studied at the ultrastructural level using gold-labeled macromolecules (bovine lactoferrin, human and bovine transferrin, bovine albumin, human low-density lipoprotein, horseradish peroxidase, and protein A). All macromolecules were ingested by the protozoan. Binding experiments showed that only bovine lactoferrin bound to the parasite surface in a process that could be inhibited by the unlabeled protein, suggesting that it binds and is internalized via receptors. Label-fracture experiments showed that the receptors were distributed in clusters that did not colocalize with intramembranous particles. Kinetics analysis of the internalization of bovine lactoferrin and horseradish peroxidase, associated with the cytochemical detection of acid phosphatase, revealed that proteins were rapidly ingested through small uncoated vesicles and delivered to acid phosphatase-containing compartments. The colocalization of gold-labeled proteins and reaction product indicative of enzyme activity was confirmed by electron spectroscopic imaging. Simultaneous incubation of cells in the presence of two proteins labeled with gold particles of different diameters showed that they were ingested through the same pathway and were concentrated into cytoplasmic vacuoles corresponding to lysosome-like organelles. These data suggest that the endocytic process in T. foetus is very rapid and that the intracellular pathway for receptor-mediated and fluid-phase endocytosis seems to be the same.

Animals↗

Rapid primary microwave-glutaraldehyde fixation preserves the plasma membrane and intracellular structures of the protozoan Tritrichomonas foetus.

Tritrichomonas foetus, a pathogenic protozoan, was used as a model to analyse microwave-stimulated fixation as a procedure of preparation of biological samples for electron microscopy of thin sections and freeze-fracture replicas. Good preservation of the protozoan structure was achieved by microwave-stimulated fixation and Epon polymerization. The membrane structure, as visualized in freeze-fracture replicas, was well preserved.

Animals↗

The structural organization of the pathogenic protozoan Tritrichomonas foetus as seen in replicas of quick frozen, freeze-fractured and deep etched cells.

The quick-freezing and freeze-etching technique was used to analyse the cytoskeleton of Tritrichomonas foetus, a pathogenic protozoan of the urogenital tract of cattle. The cytoplasm presented a network of filamentous structures interacting with each other, with the surface of the hydrogenosomes and the nuclear membrane. Two nm wide filamentous structures were found in the luminal space of the Golgi complex, connecting the two faces of each cisterna. The microtubules of the pelta-axostyle system were connected by bridges 30-40 nm long and 10 nm wide, regularly spaced with an interval of 25 nm. The costa is a structure formed by a complex array of filaments and globous structures. It seems to be connected to the recurrent flagellum through a complex network formed by 15 and 10 nm wide filaments which emerge from the peripheral region of the costa and penetrate into the surface projections of the protozoan body to which the recurrent flagellum is attached. Other filaments were seen connecting the surface of these projections with the surface of the flagellum.

Animals↗

Electron microscopic study of the effect of zinc on Tritrichomonas foetus.

At concentrations of 3.1 to 24 mM, zinc inhibits the multiplication of and kills the pathogenic protozoan Tritrichomonas foetus. Transmission electron microscopy showed that the hydrogenosome, a organelle which is involved in the metabolism of pyruvate and the site of formation of molecular hydrogen, constitutes the main site of the initial effect of zinc. The hydrogenosomal vesicle increases its electron density and dimension. Electron spectroscopy imaging and the electron energy loss spectrum showed the presence of zinc, calcium, and oxygen in the electron-dense areas of the hydrogenosome.

Animals↗

Isolation and biochemical characterization of the costa of Tritrichomonas foetus.

The costae are cytoskeletal structures found in Trichomonadidae. Both the structural organization and composition of this organelle are still unknown. In the present work we have introduced a new methodology for the costa isolation. Using sucrose density-gradient centrifugation an enriched costa fraction was obtained. Analyses by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the costa contains several proteins, with major bands corresponding to apparent molecular masses of 122, 115, 112, 93, 87, 82, 59, 52, 44, 41, 32, and 26 kDa. No significant amount of carbohydrates was detected in the costa fraction. The fractionation methodology described here has the advantage of using normal centrifugation methods and is being applied to trichomonas in the size range of Tritrichomonas foetus and Trichomonas vaginalis.

Animals↗

Cytochemical localization of enzyme markers in Tritrichomonas foetus.

Cytochemical techniques associated with transmission electron microscopy were used for the localization in Tritrichomonas foetus of enzymes used as markers of different cell structures. Reaction product indicating the presence of Mg(2+)-adenosine triphosphatase (Mg(2+)-ATPase) and 5'-nucleotidase was observed in the plasma membrane. Glucose-6-phosphatase was seen in association with the endoplasmic reticulum, revealing its organization as parallel cisternae. Thiamino-pyrophosphatase was located in the cis-most region of the Golgi complex. Acid phosphatase was found within lysosomes as well as in several cisternae of the Golgi complex, in contrast to previous observations in mammalian cells. These observations provide support for the use of enzyme markers in future studies on cell fractionation of T. foetus.

5'-Nucleotidase↗

Fibronectin- and laminin-mediated endocytic activity in the parasitic protozoa Trichomonas vaginalis and Tritrichomonas foetus.

The pathogenic protozoan Tritrichomonas foetus is able to ingest polystyrene particles with a diameter up to 1.0 micron. Trichomonas vaginalis, however, ingest particles as large as 4.4 microns in diameter. The particles are found within cytoplasmic membrane-bounded vacuoles. Morphometrical analysis showed that T. vaginalis presents a higher endocytic activity than T. foetus. Coating of the polystyrene particles with cationized ferritin increases their binding to the parasite surface but does not interfere with their ingestion. In contrast, coating with laminin significantly increased the uptake of the particles by both parasites while coating with fibronectin potentiates the ingestion of the particles only by T. foetus. These observations suggest the presence of laminin- and fibronectin-binding sites on the surface of trichomonads, an observation which is in agreement with the recent description of a receptor for laminin on the surface of trichomonads.

Animals↗

Tritrichomonas foetus: freeze-fracture cytochemistry using polymyxin B.

The general structure of Tritrichomonas foetus incubated in the presence of the peptide antibiotic polymyxin B, which interacts specifically with anionic phospholipids, was analyzed using transmission electron microscopy of thin sections, and freeze-fracture replicas. Polymyxin B induced morphological changes in the plasma membrane of the parasites with the formation of membrane blebs with a diameter varying from 65 nm to 1.5 micron. Freeze-fracture images of the membrane lining the blebs showed that their inner membrane half is smooth. However, membrane particles, with a density similar to that observed on the E face of the plasma membrane, were seen on the outer half of this membrane.

Animals↗

Localization of acetylated alpha-tubulin in Tritrichomonas foetus and Trichomonas vaginalis.

We used monoclonal antibodies specific for acetylated and nonacetylated alpha-tubulin to detect and to localize microtubules containing acetylated alpha-tubulin (stable microtubules) in the pathogenic protozoa Tritrichomonas foetus and Trichomonas vaginalis. SDS-PAGE analysis showed that tubulin is a major protein of both parasites, being enriched in cytoskeletal preparations of whole cells extracted with Triton X-100. The monoclonal antibodies, which recognize all isoforms of alpha-tubulin (B-5-1-2) and only acetylated alpha-tubulin (6-11B-1), bind to the tubulin of T. foetus and T. vaginalis as seen by immunoblotting. Tubulin-containing structures were localized using immunofluorescence microscopy and transmission electron microscopy of the whole cytoskeleton previously incubated in the presence of the anti-tubulin antibodies and a second antibody-gold complex, and then processed using the negative staining or replica techniques. The results obtained indicate that, in addition to the flagellar microtubules, those which form the peltar-axostyle system represent stable microtubules containing acetylated alpha-tubulin.

Acetylation↗

Structural analysis of the cytoskeleton of Tritrichomonas foetus.

The cytoskeleton of Tritrichomonas foetus was studied by immunofluorescence microscopy, using anti-actin, anti-filamin, anti-myosin and anti-tubulin monospecific antibodies, and by high voltage electron microscopy of Triton X-100 extracted cells. Actin, filamin and myosin were distributed throughout the cytoplasm of T. foetus. Filamin, however, is more concentrated at the cell periphery. The peltar-axostylar system could be seen in interphasic and dividing cells using anti-tubulin antibodies. High voltage electron micrographs showed the spatial distribution of the microtubules and their association with the hydrogenosomes, and the association of the costa with the recurrent flagellum.

Animals↗

Tritrichomonas foetus: ultrastructure and cytochemistry of endocytosis.

Tritrichomonas foetus ingests horseradish peroxidase, native ferritin, cationized ferritin, and 0.08 micron latex beads by a process which involves the formation of pinocytic vesicles. These vesicles fuse with each other and with lysosomes forming large vacuoles. Biochemical determinations on the ingestion of horseradish peroxidase and morphometric analysis on the ingestion of cationized ferritin covered latex beads indicated that T. foetus has high endocytic activity. The process of ingestion of the various tracers used was analyzed by transmission electron microscopy of thin sections and freeze fracture replicas.

Animals↗

Tritrichomonas foetus: cytochemical visualization of the endoplasmic reticulum-Golgi complex and lipids.

The zinc iodide-osmium tetroxide technique was used to analyze the distribution of the endoplasmic reticulum-Golgi complex system of Tritrichomonas foetus. Interconnections between the cisternae of the endoplasmic reticulum as well as between cisternae of the Golgi complex were observed. The nuclear pores, as well as fenestrations in the Golgi complex, were evident. Three to four profiles of the endoplasmic reticulum were seen in the proximal marginal lamellae, related to the attachment of the recurrent flagellum to the protozoan body. No reaction product was seen in the costae, microtubules, glycogen particles, or hydrogenosomes. Imidazole-buffered osmium tetroxide solution was used to visualize lipids. Electron-dense materials were seen either free in the cytoplasm or within membrane-bounded cytoplasmic vesicles. A high contrast of some membranes, mainly of those which enclosed the hydrogenosomes, was observed in unstained sections.

Animals↗