Biomedical subjects

C Argüello

Publications and source records attributed to C Argüello.

At least 19 recordsLinked to original sources

PubMed2004

Surface properties and in vitro cytopathic effect of various strains of Trichomonas vaginalis.

The in vitro cytopathic effect of four strains of Trichomonas vaginalis on cultured epithelial monolayers was analyzed through electrophysiology and electron microscopy. Interaction of trichomonads of two virulent strains (GT-10 and GT-13) with cultured MDCK cell monolayers mounted in Ussing chambers produced a rapid decrease in transepithelial resistance to less than 30% of control values after only 15 min. By 30 min the electrical resistance was practically abolished by the virulent parasites. In contrast, of two attenuated strains of trichomonads (GT-3 and GT-7) analyzed under similar conditions, GT-3 trophozoites required 180 min to reduce transepithelial resistance to 9% of control values, while monolayers in contact with GT-7 parasites still showed 28% of control values at this time of incubation. Sequential scanning electron microscopy confirmed the much faster and widespread cytopathic effect of virulent parasites. In contrast, the slow lytic process produced by attenuated trophozoites was reduced to focal areas of direct contact with epithelial cells. Another difference was found by measurement of the surface charge of the four strains of T. vaginalis by means of cell microelectrophoresis. While the two virulent strains showed a negative surface charge, the two attenuated strains had no detectable surface charge at neutral pH. When parasites were incubated with cationized ferritin and studied with transmission electron microscopy the surface of virulent trichomonads appeared heavily labeled, whereas the surface of attenuated parasites had only sparse and irregular ferritin binding.

Animals↗

PubMed2000

Isolation and analysis of a new developmentally regulated gene from amastigotes of Leishmania mexicana mexicana.

Leishmania differentiates from the promastigote to the amastigote stage during its digenetic life cycle. Characterization of the developmentally regulated genes during that process would help to elucidate the mechanisms of gene regulation. In this study, specific fragments of mRNAs from the amastigote stage of L. mexicana mexicana were discriminated from those of the promastigote and metacyclic stages by differential display. This technique combined with spliced-leader polymerase chain reaction allowed isolation of the complete gene VG7A5. The sequence of this gene did not align with any published L. mexicana sequence. More than one copy of this gene was identified in the genome by Southern-blot analysis and was transcribed exclusively in the amastigote stage. At 20 bp upstream from the splice AG site it has a trans-splicing polypyrimidine tract. The gene encodes the subcellular localization motifs 5'-GGACT and AAGCT-3' in the 3' untranslated region of the mRNA. The open reading frame of the gene VG7A5 predicts a polypeptide of 587 amino acid residues that has a KGRR amidation motif near its carboxyl terminus, suggesting that in the mammalian host this protein may be involved in the process of acute inflammation.

5' Untranslated Regions↗

PubMed1998

Use of microscopical techniques in the study of human chondrocytes from osteoarthritic cartilage: an overview.

Several microscopical techniques, such as high resolution light microscopy, Normaski microscopy, laser confocal and transmission electron microscopy, were used in a correlative morphological study of human osteoarthritic (OA) cartilage. Emphasis was made on the characterization of chondrocytes heterogeneity observed in this tissue. Novel findings were assessed in the morphological and immunocytological study of the chondrocytes organized in aggregates or "clones" typical of this degenerative disease, consisting of the modification of certain elements of the cytoskeleton that influence changes in the cell shape. Also, the presence of cilia and centrioles found in certain cell raised the question if chondrocytes are able to move and regroup as an alternative mechanism to mitosis in the formation of cell clusters or "clones." The presence of two types of secretory chondrocytes was observed and discussed. The use of a correlative approach of several microscopical techniques in a systematic morphological and immunocytological characterization of chondrocyte population within the fibrillated and nonfibrillated human osteoarthritic cartilage gave complementary information that could be important for a better understanding of the histopathogenesis of OA.

Cartilage, Articular↗

PubMed1998

Myocardial interatrial septum loses its epithelial organization by mesenchymal influence. Structural and ultrastructural study.

During atrial septation, the septum primum fuses with the atrioventricular endocardial cushions and myocardial-mesenchymal interactions occur. In order to evaluate the cellular events that take place during this particular interaction a structural, ultrastructural and histochemical study was performed. Our findings indicate that from the fourth day of development, the distal myocardium of the interatrial septum, which interacts with mesenchymal tissue, loses its appearance of an epithelial sheet and becomes a loosely organized tissue. The distal myocytes of the interatrial septum which get progressively separated show features of migratory cells, the final localization of which is the mesenchymal tissue of the adjacent endocardial cushions. These tissue changes involve basal membrane disruption, reduction in the number of desmosomes and intercalated discs with the subsequent appearance of large intercellular spaces between myocytes, myofibrillar disarrangement and acquisition by myocytes of a secretory phenotype characterized by numerous cytoplasmic vesicles. These events occur in a similar way in the atrioventricular canal, where a myocardial-mesenchymal interaction also occurs. In both regions the mesenchymal endocardial cushions and its associated extracellular matrix seem to direct the dissociation of the myocardial tissue and the subsequent migratory cellular behaviour of the interacting myocytes. This is an interesting, and little known, example of a cellular phenotypic transformation during cardiac development.

Animals↗

PubMed1997

Leishmania mexicana: binding of promastigotes to type I collagen.

During leishmania infection, parasites are inoculated to the human host through the bite of a sandfly vector into the dermis, where they first interact with tissue components, cells and extracellular matrix molecules. Since collagen is the most abundant component of the skin matrix, we investigated whether there is a specific interaction of Leishmania mexicana promastigotes with this host component. Promastigotes were able to attach to collagen fibrils and move through the matrix of mouse skin sections and to penetrate easily into a type I collagen gel. Denatured type I collagen coated beads (Cytodex 3) readily bound to the parasite surface. The interaction of promastigotes with type I collagen was dose dependent and saturable and was competitively and specifically inhibited with increasing concentrations of gelatin. Biotin-labeled parasite surface molecules were able to associate with both denatured collagen from microcarriers and native type I collagen from bovine kidney. It is suggested that the presence of parasite cell membrane receptors to collagen may confer a specific tropism for the skin, where collagen is the most abundant component of the matrix.

Animals↗

PubMed1997

Effects of heparin on the electrophysiological and mechanical properties of early embryonic chick hearts.

The effect of heparin (1-200 microg/ml) on several functional properties of 4-day-old embryonic hearts was studied. Intracellular recordings either from cells of the atrium, the atrio-ventricular canal or the ventricle were made on the isolated and perfused hearts. In all impaled myocardial cells, heparin produced an increment in the maximal rate of rise and amplitude of the action potentials and the duration was slightly diminished. In addition, heparin provoked a reduction in the spontaneous beating frequency, while the electrical excitability and the conduction velocity of the atrium and the ventricle were increased. The measurements of atrio-ventricular latency revealed that it was diminished, i.e. the propagation of impulses through the atrio-ventricular canal was improved. Simultaneously with these changes, heparin incremented the amplitude of contraction of either the atrium or the ventricle importantly.

Animals↗

PubMed1996

Transient outward K+ channels in vesicles derived from frog skeletal muscle plasma membranes.

Whole-cell voltage-clamp experiments were performed in vesicles derived from frog skeletal muscle plasma membranes. Capacitance measurements showed that these vesicles lack invaginations. In solutions containing K+, transient outward currents with reversal potentials close to EK were recorded with a maximum potassium conductance of 0.3 mS/cm2. These currents inactivated in a voltage-dependent manner with a time constant of decay that reached a limiting value of 26 ms at large depolarizations. The steady-state inactivation reached half-maximum values at -66 mV. Transient currents were completely blocked with 5 mM 4-aminopyridine. Single-channel recordings made in inside-out excised patches from the vesicles had ensemble averages with characteristics similar to those of the macroscopic currents, although with significantly faster inactivation time constants. The single-channel chord conductance was 21 pS when the pipette and bath solutions contained 2.5 mM and 120 mM KCl, respectively. It is concluded that these vesicles contain potassium channels that are very similar to A channels found in neurons and other cells.

Animals↗

PubMed1996

Variability in the cell phenotype of aggregates or "clones" of human osteoarthritic cartilage. A case report.

Human samples of articular cartilage from the knee of a clinically classified osteoarthritic patient, assessed by arthroscopy as part of the surgical treatment was studied by light and transmission electron microscopy. This particular case differed from others already reported in the variability of cell phenotype within the aggregates or "clones" frequently present in the osteoarthritic cartilage. The most common morphology of "clonal" cells forming the aggregates were large and rounded with an euchromatic nucleus. The cytoplasm was characterized by the presence of alternately clear and dense sites. At the ultrastructural level it was seen that the clear sites were formed by disrupted intermediates filaments and small particles, and that the dense sites were constituted by the segregation of different organelles of the chondrocytes. In addition, there were atypical aggregates composed only by secretory cells or by degenerating chondrocytes. Furthermore, a complex structure consisting of a very large cell inside a giant lacunae delimited by electron-dense material with small vesicles is described as a novel finding. The variability in the chondrocyte phenotype of the aggregates described here could be an indication of a better prognosis; nevertheless, the follow-up of the evolution of this patient is needed in order to know the final outcome.

Cartilage, Articular↗

PubMed1994

Transbilayer diffusion of divalent cations into liposomes mediated by lipidic particles of phosphatidate.

Liposomes formed from egg-yolk phosphatidylcholine:egg-yolk phosphatidate (molar ratio 2:1) containing pBR322 DNA and DNase I were induced to form, with divalent cations, bilayer/nonbilayer phase transitions of phosphatidate which allowed cation diffusion into liposomes; then cation diffusion was measured by the activation of the hydrolysis of DNase I on DNA. The formation of phosphatidate transitions on liposomes was demonstrated by freeze-fracture and 31P NMR, and a direct correlation between the formation of phosphatidate transitions and the transbilayer diffusion of cations was found: only Ca2+ and Mn2+, which induce phase transitions, were able to penetrate liposomes and triggered the DNase I activity; in addition, Ca2+ at higher concentrations (10 mM) caused fusion of liposomes, whereas Mn2+ did not, suggesting that transitions induced by Mn2+ participated only in the diffusion of this ion; furthermore, Mg2+ neither formed phase transitions nor triggered the enzymatic activity. The liposomes studied represent more dynamic structures that can form phosphatidate structures involved in both (1) the interchange of divalent cations with the surroundings, thereby modulating encapsulated enzymes, and (2) the fusion of lipid vesicles probably implicated in the enrichment of liposomal content in the early Precambian Earth.

Cations↗

PubMed1993

Leishmania braziliensis promastigotes and amastigotes interact differently with host macrophages.

In vitro and in vivo ultrastructral studies reveal that the parasite entrance into the macrophage occurs by phagocytosis. The early stage of phagocytosis exhibited different ultrastructural characteristics in both forms of the parasite. Long and prominent projections from peritoneal exudate macrophages made focal contacts with the promastigote surface. The amastigotes, in turn, laid on cup-shaped extensions of the macrophage membrane. Later stages of the phagocytosis are characterized by progressive and complete engulfment of both promastigotes and amastigotes.

Animals↗

PubMed1992

Sparganum proliferum: an overview of its structure and ultrastructure.

A detailed study of the structure and ultrastructure of Sparganum proliferum was made possible for the first time thanks to the successful in vitro and in vivo maintenance of this rare parasite. Although S. proliferum exhibits many of the classical tegumental and parenchymal structures previously described for other larval cestodes, these are either arranged in a distinct fashion or, in some cases, may be completely different. Among the latter and of special interest are the single or multiple parenchymal cavities, surrounded by tegument, which in some instances appear to act as a primitive digestive tract.

Animals↗

PubMed1992

Structural organization of chromatin during the cell cycle of Entamoeba histolytica trophozoites.

The nuclear division of E. histolytica trophozoites was analyzed by using specific stains for DNA, with the aim to define the sequential changes of chromatin during its life cycle. Furthermore, we characterized the internal structural arrangements of microtubules in the microtubular organizing center (MTOC) and determined the number of chromosomes and its association with the spindle. The MTOC is formed by multiple microtubule-nucleating centers, that are involved in the displacement of DNA during nuclear division. We found the existence of a single MTOC in one pole of the nucleus at early anaphase. Our results lead us to propose a new hypothesis in which it is suggested that metaphase corresponds to the arrangement of condensed DNA bodies, or "chromosomes" around the MTOC and, through the assembly of microtubules, one set of uncondensed chromatin is displaced to the opposite pole of the nucleus, while the other remains condensed and associated to the original MTOC. We observed six chromosomes in our preparations, corroborating previous observations (2,3). Whether or not a new MTOC is formed during nuclear division remains to be clarified.

Animals↗

PubMed1990

Diffusion of Mn2+ ions into liposomes mediated by phosphatidate and monitored by the activation of an encapsulated enzymatic system.

Transbilayer diffusion of Mn2+ ions occurred in liposomes formed from dipalmitoyl-phosphatidylcholine or egg-yolk phosphatidylcholine and egg-yolk phosphatidate (molar ratio 2:1) containing DNA and DNase I within their aqueous compartments. Cation diffusion was demonstrated by the hydrolytic activity of DNase I, activated by the Mn2+ ions that diffused into the vesicles, and this was confirmed by light scattering. Phosphatidate, a cone-shaped lipid which has been synthesized under simulated prebiotic conditions, was necessary for cation diffusion across the liposome membranes. Such liposomes represent a simple precellular system that interchanges cations with the surroundings and provides a microenvironment for enzymatic reactions, as evidenced by the hydrolysis of DNA by DNase I inside these closed lipid compartments.

Cations↗

PubMed1990

[Identification and localization of molecules which intervene in the interaction between Entamoeba histolytica and epithelial cells].

During the initial stages of host-parasite interaction between Entamoeba histolytica and its target cell, superficial molecules of both cells participate. The purpose of this study has been the study of the parasite's proteins which are involved in the process of adhesion, as well as localize the superficial sites of the host which bind with the amebic proteins. Antibodies directed against adhesin 112 kDa, and other amebic adhesins, specifically inhibited the destruction of the monolayers of the MDCK epithelial cell line. Soluble proteins of MDCK cells recognized was separated by electrophoresis and transferred to nitrocellulose paper, the amebic proteins of 112, 90 and 45 to 50 kDa. In a similar fashion, membrane proteins of MDCK cells marked with 125[I] specifically recognized amebic proteins of 112, 90, 45 and 32 kDa, as well as others of higher molecular wight. The localization of the binding sites of the E. histolytica molecules on semi-confluent monolayers of MDCK cells, was brought about by immunofluorescence using antibodies against the parasite's total proteins or against adhesins. For electron microscopy antibodies labelled with peroxidase were used. The amebic proteins were mainly found on the periphery of free cells, land in the intercellular cell junctions of monolayers.

Animals↗

PubMed1989

Development of the aorta in the chick embryo: structural and ultrastructural study.

A structural and ultrastructural study was designed to analyze systematically the cellular events which take place in the aortic wall between days 7 and 21 of chick embryo development. Between days 7 and 18, increase in total diameter, number of cell layers, and aortic wall thickness are highly correlated, whereas between days 18 and 21 the total diameter increase is correlated mainly with an increase in vessel lumen diameter. Cell layers of smooth muscle cells showing an immature or synthetic phenotype arise from progressive association and organization of mesenchymal cells originated from an endothelial activation process in which a hyaluronic acid-rich extracellular matrix seems to be involved. It is suggested that the process of endothelial activation takes place between days 7 and 18 of embryonic development provided that within that period the typical cellular events which are involved in such a process take place (hypertrophy, reorientation, invagination, mitotic activity, acquisition of migratory appendages, endothelial detachment and incorporation into adjacent spaces). This endothelial activation has been recognized as a selective multiphasic process required for the transition of endothelial cells into mesenchyma.

Animals↗

PubMed1988

Culture of proliferating and differentiating fat-storing cells in 3T3-conditioned medium.

There is growing evidence suggesting that hepatic fat-storing cells (FSC) or Ito cells have an important function in vitamin A storage and metabolism and in the synthesis of connective tissue components in normal liver and during fibrogenesis. The purified FSC acquire a fibroblastic morphology and their vitamin A content decreases in culture. We cultivated cells under in vitro conditions that allowed the expression of FSC morphological and functional characteristics for 3-4 weeks of primary culture. Cells were isolated from rat liver by the collagenase-perfusion method without further purification and cultured with 3T3-conditioned medium, which seemed to stimulate the selective proliferation of the FSC. After 8-10 days, round and stellate cells grew actively from a few precursor cells in the primary culture and were not subcultivated; the stellate cells had the ability to become round and vice versa and were highly motile. The cells had intracytoplasmic lipid droplets, a well developed rough endoplasmic reticulum, Golgi complex, numerous vesicles filled with electron-dense material, and extracellular matrix (ECM) components on their surface. Both stellate and round cells showed the presence of desmin by immunofluorescence and vitamin A autofluorescence, but lacked peroxidase activity. The culture conditions we describe allowed the selective proliferation of cells with morphological and functional characteristics of the FSC in the normal liver, raising the possibility of studying FSC proliferation and differentiation.

Animals↗

PubMed1988

The early development of the atrioventricular node and bundle of His in the embryonic chick heart. An electrophysiological and morphological study.

The development of the atrioventricular node and bundle of His of embryonic chick hearts was studied by electrophysiological and morphological techniques. The dorsal wall of the AV canal and the interatrial septum were explored to determine if they contribute to the formation of the AV node and bundle of His. The resting membrane and action potentials of the interatrial septum cells were systematically analyzed and found to undergo progressive differentiation with development. The earliest identification of the AV node and upper bundle of His group of cells was achieved at 5 1/2-6 days of development by the electrical recording of their corresponding characteristic action potentials, from a circumscribed area located in the lowest and dorsal segment of the interatrial septum. The morphological and anatomical characterization of the cells was made following electrical recording and labelling with charcoal particles. The earlier AV node and bundle of His responses had similar characteristics to those of the adult heart. It is concluded that the AV node and upper bundle of His cells derive from the low interatrial septum. The possibility that AV canal cells contribute to this event was discarded. The functional relationship of the Av node and bundle of His with other cardiac tissues during the early development of the heart is discussed.

Animals↗

PubMed1987

Influence of the mesenchymal microenvironment on myocardial and endocardial cell behaviour in experimental interaction with chick limb mesenchyme.

In an attempt to clarify the possible influence of the mesenchymal microenvironment in the differentiation of myocardial and endocardial cells, an "in vivo" transplantation experiment was performed in which the ventricular region of the heart (chick or quail) was placed in close association to the mesenchyme of the anterior chick limb to create an experimental interaction between myocardium and foreign mesenchyme. The results showed that after 48 h of tissue interactions the ventricular myocardium is incorporated into the mesenchyme of the anterior chick limb, changing its organization and cytological appearance. The myocytes tend to dissociate, exhibiting a less organized myofibrillar pattern. In addition, abundant extracellular matrix components made up of granular and fibrillar material were observed associated with the myocardial and the mesenchymal cell membranes as well as distributed in their surrounding microenvironment. The endocardium became discontinuous, due to detachment of the cells and emitted multiple pseudopodia and filopodia. These observations indicate that the mesenchyme from the anterior chick limb modifies the cellular behaviour and organization of the neighbouring myocardium and endocardium with which it interacts. We suggest that this might occur through participation of extracellular matrix components such as glycosaminoglycans, fibronectin and collagen which are known to act as macromolecular mediators in cell to cell interactions, cell migration and differentiation.

Animals↗