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Biomedical subjects

J M Senet

Publications and source records attributed to J M Senet.

At least 37 records · Page 2Linked to original sources

Binding of mouse fibrinogen to Candida albicans in vivo.

Binding of fibrinogen to various Candida albicans strains has been investigated by immunofluorescence microscopy on kidney sections of experimentally infected mice. Fibrinogen appeared to bind to both mycelium and blastospores in situ whereas previous studies, carried out in vitro, have shown fibrinogen binding to mycelial elements only.

Animals↗

Fungal cell adhesion molecules in Candida albicans.

Adherence of Candida albicans to host tissues is considered a crucial step in the pathogenesis of candidiasis. Using in vitro assays, it was demonstrated that the yeast-mycelium transition was an important phenomenon in the acquisition of adhesive properties. Proteins with MWs of 60, 68, 200 and greater than 200 kDa seemed to be involved in germ tube adherence to plastic surfaces. Likewise, recent investigations have revealed that C. albicans expresses on its surface receptors which interact with a wide variety of host proteins, particularly some extracellular matrix components like fibronectin, laminin and collagen. Plasmatic components, such as fibrinogen, iC3b and C3d, have also been proposed as mediators of adherence of C. albicans. Thus, by their reaction with laminin, fibrinogen and C3d, the mannoproteins of 68 and 60 kDa demonstrated multiple biological activities. Proteins of similar MWs were detected as C3d and iC3b receptors, the latter showing similarities with the neutrophil CR3. Based upon the antigenic, structural and functional homologies between the candidal receptors and mammalian integrins, it was postulated that these fungal cell adhesion molecules (F-CAM) are members of the integrin family. Interactions with host proteins and molecular mimicry of mammalian adhesion molecules may be a fertile area for further research.

Basement Membrane↗

Specific interaction between Listeria monocytogenes and glycosylated albumins.

We have previously shown that Listeria monocytogenes serovar 1/2b can bind strongly to bovine albumin (BA) glycosylated by glucosamine or fucosylamine with about 20 to 30 carbohydrate residues per albumin molecule. We now show that the binding is time-dependent, reversible, saturable and specific. The two glycosylated compounds inhibit each other competitively. Scatchard analysis showed that about 100 molecules of BA-glucosamide (heptameric configuration) and 14,300 molecules of BA-fucosylamide (monomeric configuration) bound per bacterial cell. The apparent dissociation constants for BA-glucosamide and BA-fucosylamide were found to be 3.9 x 10(-14) M and 3.5 x 10(-13) M, respectively.

Albumins↗

Reaction of Candida albicans cells of different morphology index with monoclonal antibodies specific for the hyphal form.

Two monoclonal antibodies (MAbs), 3D9 with reported specificity for Candida albicans hyphae, and 3B7 with reported specificity for morphological forms of C. albicans found in vivo, were tested by indirect immunofluorescence with C. albicans cells that were grown in 12 different environments (four different culture media incubated at various temperatures) and whose cellular morphology was estimated in terms of morphology index (Mi). Both MAbs reacted strongly with cells with Mi greater than 3.0, i.e., with pseudohyphal and hyphal forms, but in Eagle's medium at 26 degrees C and in a modified Sabouraud's broth medium at 30 degrees C, some reactivity was also found with cells of lower Mi (i.e., yeast forms). Therefore, it was concluded that the hyphal phenotype and the epitopes reactive with the MAbs were co-expressed but that the epitopes could also be expressed independently of the hyphal phenotype. The results confirm the propensity of C. albicans for variation of its surface antigenic composition.

Antibodies, Fungal↗

Specific binding of neoglycoproteins to Toxoplasma gondii tachyzoites.

Several studies have shown that protozoa bind to glycoproteins or neoglycoproteins. Here we report that Toxoplasma gondii binds strongly to bovine serum albumin-glucosamide. The binding was rapid, time dependent, partially reversible, saturable, and specific. Scatchard analysis showed about 40,000 molecules of bovine serum albumin-glucosamide per toxoplasma cell. The apparent dissociation constant was found to be 4.46 x 10(-8) M.

Animals↗

[Q fever and fetal death in utero. Two cases].

Some cases of late abortion occurring after a Coxiella burneti infection, more often with a chronic evolution, have already been mentioned in the literature. We reported here two cases of early abortion, contemporaneous of an acute infection due to C. burneti. Two patients, after a contact, before and at the beginning of the pregnancy, with an animal susceptible to contaminate human beings by C. burneti, presented no clinical symptom characteristic of Q fever. The fetal death for the two cases was found out at the 17th week of amenorrhoea. All the investigations in order to search for an abortion etiology remained negative. Only, the specific serologies showed an acute infection due to C. burneti.

Adult↗

Surface Listeria monocytogenes carbohydrate-binding components revealed by agglutination with neoglycoproteins.

Carbohydrate-binding components were shown to be present at the surface of Listeria monocytogenes by means of a panel of neoglycoproteins using direct agglutination. These lectin-like components bind on neoglycoproteins bearing D-glucosamine, L-fucosylamine, or para-amino-phenyl-alpha-D-mannopyrannoside residues. The interactions were inhibited by the carbohydrate moieties specific to the neoglycoproteins. The protein nature of the lectin-like components of L. monocytogenes was ascertained by the loss of carbohydrate-binding capacity following protease treatment.

Agglutination Tests↗

Structures involved in the binding of human fibrinogen to Candida albicans germ tubes.

Recent evidence for the interaction between human fibrinogen and Candida albicans germ tubes have led us to attempt to characterize the structures involved. Using 125I-radiolabeled proteins, fibrinogen purified by affinity chromatography and its plasmin degradation products, the binding sites on the fibrinogen molecule appeared to be located specifically in the D-domain. Conversely to the fibrinogen and the fragment D, radiolabeled fragment E, however, did not interact with cells. The binding was time-dependent, saturable and reversible. Scatchard analysis of the data obtained revealed an average of 6000 binding sites per germ tube with dissociation constant (Kd) of 5.2 X 10(-8) M. No potent competition was observed for a range of different proteins and carbohydrates. Fibrinogen fragment D binding proteins were identified using a dithiothreitol-iodoacetamide extract of the fungus. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, one component of 68 kDa was detected. Thus, the presence of fibrinogen binding proteins specifically localized on the cell wall surface of C. albicans germ tubes may constitute one of the factors involved in the development of candidosis.

Binding Sites↗

Laminin receptors on Candida albicans germ tubes.

Recent evidence for the role of laminin in cell adhesion and in the pathogenesis of several bacterial infections has led us to investigate the existence of receptors for this extracellular matrix component in Candida albicans. At first, immunofluorescence demonstrated the presence of laminin-binding sites at the surface of germ tubes. Electron microscopy confirmed this result and permitted precise localization of the binding sites on the outermost fibrillar layer of the germ tube cell wall. By using 125I-radiolabeled laminin, the binding was shown to be saturable and specific, hence demonstrating characteristics of true receptors. Analysis of the data by the Scatchard equation indicated that there were about 8,000 binding sites per cell, with a dissociation constant (Kd) of 1.3 x 10(-9) M. Binding was inhibited by prior heating or trypsinization of cells. Furthermore, of the different proteins and carbohydrates tested in competition experiments, only fibrinogen greatly reduced the laminin binding. Finally, dithiothreitol and iodoacetamide treatment of germ tubes allowed us to identify the laminin receptors through analysis of this extract by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting. Two components, of 68 kilodaltons and a doublet of 60 and 62 kilodaltons, were detected. Thus, C. albicans possesses germ tube-specific surface receptors for laminin which could mediate its attachment to basement membranes and so contribute to the establishment of candidiasis.

Candida albicans↗

Evaluation of a new bicolored latex agglutination test for immunological diagnosis of hepatic amoebiasis.

A new bicolored latex agglutination amoeba test (BLA) for detection of antibodies against Entamoeba histolytica was evaluated for its practicability and diagnostic sensitivity and specificity. BLA is rapid (5 min) and simple to perform. It requires only 20 microliters of a 1/3-diluted serum, 17 microliters of reagent, and a glass slide. Reading of the test is easy because a positive result shows a green spot with a red surrounding edge. This bicolored pattern is easily distinguishable from the negative test result, which shows a homogeneous dark-brown spot. By using serum samples from 348 individuals, BLA was compared with immunofluorescence assay, indirect hemagglutination, and counterimmunoelectrophoresis. Sensitivity, specificity, efficiency, and positive and negative predictive values of the four methods were almost identical. The results of this study indicate that BLA could be very useful both as a screening method for the diagnosis of invasive amoebiasis and for epidemiological purposes.

Amebiasis↗

Adherence of Candida albicans germ tubes to plastic: ultrastructural and molecular studies of fibrillar adhesins.

Germ tubes of Candida albicans produced an additional fibrillar surface layer responsible for enhanced adherence to plastic. The correlation between germination of C. albicans and adherence of germ tubes to a plastic matrix led us to consider the existence of germ tube-specific adhesive components involved in the attachment process. Using concanavalin A-sensitized latex microspheres, we first detected extracellular molecules on the plastic surface after removal of the adherent germ tubes. Electron microscopy confirmed that fibrils of the germ tube involved in cell-substratum interconnections were retained on the plastic surface. Cytochemistry with concanavalin A-gold labeling demonstrated that these fibrillar structures contained mannoproteins. Dithiothreitol and iodoacetamide treatment of washed plastic allowed us to further characterize these fibrillar adhesins. Through analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two components with molecular weights (MWs) of 68,000 and 60,000 were detected on the plastic surface. The 68,000-MW component appeared to be one of the major constituents of the germ tube surface layers. Biosynthetic labeling experiments performed with L-[35S]methionine revealed two additional proteins: a high-MW component (greater than 200,000), and a 200,000-MW component. These four proteins, strongly labeled on the plastic surface and on the germ tube cell wall layers, were in contrast slightly labeled or even nonidentified in the culture supernatant, suggesting their involvement in germ tube adherence.

Adhesiveness↗

Binding of fibrinogen to the pathogenic Aspergillus species.

The binding of human fibrinogen to the pathogenic aspergilli was investigated in vitro by different procedures using either fibrinogen in solution or fixed, insolubilized fibrinogen. Binding of fibrinogen was detected at the surface of hyphae and conidia by an immunofluorescence assay. Ultrastructural localization of the binding sites was visualized with fibrinogen-sensitized gold particles. The labelling was restricted to the outer cell wall layer of the 'smooth' walled conidia. Quantitative analysis of the binding carried out with 125I-labelled human fibrinogen on 33 species belonging to different groups (opportunistic fungi, strictly saprophytic or phytopathogenic fungi and dermatophytes or related species) clearly demonstrated that among all the fungi tested, only the pathogenic aspergilli significantly bound fibrinogen. The average amount of fibrinogen bound to individual conidia was also quantified. Binding was greater to Aspergillus niger (5-fold), Aspergillus fumigatus (2.5-fold) and Aspergillus flavus conidia (2.3-fold) than to Aspergillus terreus conidia. The results suggest that fibrinogen binding could contribute to the pathogenesis of aspergillosis.

Aspergillus↗

Characterization of binding of human fibrinogen to the surface of germ-tubes and mycelium of candida albicans.

The binding of human fibrinogen to germ-tubes and mycelium of Candida albicans, forms usually found in infected tissues, was studied in vitro by an immunofluorescence assay. Binding was quantified by using 125I-labelled fibrinogen. The degree of binding differed according to the morphological form of the fungus. Binding relative to that of the yeast form was greater for mycelium (12-fold) than for germ-tube (7.7-fold). Pretreatment of yeasts with fragments D and E (terminal degradation products of fibrinogen) before fibrinogen binding showed that fragment D possessed a higher affinity for C. albicans than fragment E. Binding of fibrinogen was diminished when C. albicans was pretreated with 2-mercaptoethanol alone or in combination with pronase, or pretreated with alpha-mannosidase or trypsin. Binding was not decreased by pretreatment with pronase alone or chitinase. Inhibition experiments using C. albicans dialysed culture filtrate, C. albicans mannan, chitin, sugars or amino sugars were done by preabsorbing the fibrinogen with each of the above mentioned compounds. C. albicans dialysed culture filtrate inhibited the binding more strongly than C. albicans mannan. However, fibrinogen binding to C. albicans was not significantly reduced by mannose, several other sugars or chitin. These studies demonstrate the existence of a fibrinogen-binding factor (FBF) strongly associated with the surface of germ-tube and filamentous forms of C. albicans, and indicate a possible role for FBF in the pathogenicity of C. albicans.

Candida albicans↗

Binding of human fibrinogen to Candida albicans in vitro: a preliminary study.

The binding of human fibrinogen to different morphological forms of Candida albicans (blastoconidia, germ tubes and hyphae) was investigated by immunofluorescence and electron microscopy. Fibrinogen appeared to bind more strongly to hyphae than to yeast forms, and was detected on the cell wall coat. These observations suggest a possible role for fibrinogen in fungal colonization processes in vivo.

Candida albicans↗

[Demonstration of host-parasite common antigens in hydatidosis].

By means of gel agar diffusion techniques (Ouchterlony and immunoelectrophoresis), the authors have demonstrated in hydatid cyst fluid and membrane of E. granulosus, the presence of, at least, one fraction showing a human albumin specificity. This is not a contamination from the host and must be synthesized by the parasite itself. The significance of such a fraction is discussed.

Animals↗