Pleural effusion and toxocariasis.
The case history is described of a woman who presented with bilateral pleural effusions caused by Toxocara canis infestation. The condition responded rapidly to treatment.
Biomedical subjects
Publications and source records attributed to D Chabasse.
The case history is described of a woman who presented with bilateral pleural effusions caused by Toxocara canis infestation. The condition responded rapidly to treatment.
Cercarial dermatitis is a parasitic impasse that has worldwide distribution. The condition manifests itself as a highly pruriginous skin rash and is due to penetration of the dermis by larval stages (furcocercariae) of avian trematodes. Many species may be responsible for this disease. In Europe the genus Trichobilharzia is widely represented, in particular by the species T. ocellata; the definitive host is the duck (Anas platyrhinchos); the intermediate hosts are snails of the genus Lymnea (L. ovata or L. stagnalis). In France, cases of cercarial dermatitis were reported in June and July 1994 to the health authorities of three départements in the Pays de la Loire Region (western France). The epidemiological situation, common to the three maintained ponds that were concerned, is as follows: high level of eutrophication of the sites, colonization of the ponds by L. ovata, and settlement by many duck colonies. The simultaneous occurrence of these three phenomena, combined with long hours of sunshine in the summer, is responsible for most of the foci of cercarial dermatitis recently described in Europe. Control of this condition is difficult, requiring strict maintenance of bodies of water and if necessary the use of molluscicides such as niclosamide. The use of praziquantel in baits for treating the definitive hosts appears to interrupt the natural cycle of the avian Schistosomatidae. In the light of the observations reported here and the analysis of recent publications, cercarial dermatitis may be regarded as an emerging disease. Its public health impact needs to be evaluated at the global level.
Fungi of the order Mucorales determine various infections involving principally the respiratory tract. In spite of their medical importance, little is known about their mechanisms of adherence to the host tissues. Thus we have attempted to define the morphological stages involved in the adherence process of Rhizopus oryzae which is the main causative agent of mucormycoses. The study of the kinetics of germination and adherence to plastic revealed that attachment occurred prior to germination and decreased dramatically with germ tube formation. This correlates with important modifications of the cell wall of the fungus with respect to both carbohydrate composition and distribution of anionic sites. Moreover, the attachment of spores to extracellular matrix components immobilized onto wells of polystyrene microtiter plates has been investigated. Spores adhered readily to immobilized laminin or type IV collagen, but not to fibronectin or the glycosaminoglycans. Attachment to laminin and collagen was dose-dependent and specific. Adhesion was not inhibited by the different carbohydrates tested, suggesting that a lectin was not involved in these interactions. Finally, immunofluorescence revealed that laminin and type IV collagen interacted exclusively with spores and mother cells of germ tubes. Thus, the recognition of laminin or collagen by spores may participate in their adherence to epithelial basement membranes exposed after epithelial tissue damage which frequently accompanies the predisposing factors for mucormycoses.
In order to define the prevalence of Candida ciferrii in onychomycosis, the fungal biota associated with toe nail onyxis was examined in 50 elderly patients with trophic disorders of the legs and in 220 patients without clinical evidence of trophic disorders. Candida ciferrii was more frequent in the first group of patients since it was recovered from 24% of these patients, whereas its prevalence was only 1.4% in the control group. Moreover, the positivity of the direct examination of toe nail scrapings, the absence of any other associated pathogens, and the repeated isolation of this yeast species for some of the patients confirmed its pathogenicity.
The development of aspergillosis in an immunodeficient host depends on interactions between fungal and host components. The recognition by Aspergillus fumigatus of fibrinogen and laminin, and the secretion of extracellular proteinases and ribonucleotoxin have been suggested to mediate adherence to mucosal surfaces and subsequently to bring about host-tissue invasion.
Culture conditions that lead to swelling and germination dramatically influence cell surface characteristics and properties of Aspergillus fumigatus conidia. Conidial adherence to polystyrene and agglutination markedly increased during swelling, in a time-dependent manner. Agglutination appeared to be sensitive to cycloheximide and calcium. Removal of cell wall polysaccharides by lyticase or sodium metaperiodate suppressed agglutination of conidia. Proteinase K weakly decreased it whereas dithiothreitol strongly dispersed the cells. These observations suggest that both cell surface carbohydrates and proteins are involved in the agglutination process. Electron microscopic observations demonstrated that the cell wall of conidia was subject to some rearrangements during swelling, involving degradation and loss of the external convoluted layer, and subsequent exposure of underlying ligands. This was confirmed using lectins labelled with gold or fluorescein isothiocyanate, which showed that some carbohydrates, particularly those acting as ligands for peanut agglutinin, are largely exposed during the process. Finally, SDS-PAGE revealed major protein changes between resting and swollen conidia. We conclude that the ability of A. fumigatus conidia to aggregate correlates with an increase in adherence and biochemical reorganization of the cell wall.
Toxocara canis is a common parasite in puppies. The danger to human health has not been properly established. We estimated the current incidence of this pathogen in two western districts of France, Loire-Atlantique and Maine et Loire. Blood samples from 1836 eosino-philic patients were collected and tested by an enzyme-linked immunosorbent assay (ELISA) excretory-secretory Toxocara antigen test. We obtained positive results in 22% of the cases and highly positive results in 7%. The ELISA data seemed to be age-dependent, with older patients having more positive results (P < 0.0001). The interlaboratory distribution of positive test results was statistically significantly different (P < 0.0001), suggesting regional sources. The main clinical expressions of toxocariasis were: asthenia, gastric pain, and pulmonary disease. Individual and collective surveys of this zoonotic disease need to be carried out.
Interaction between Aspergillus fumigatus conidia and different proteins known to mediate the attachment of malignant tumor cells or microorganisms to the host tissues was studied in vitro. Flow cytometry using fluorescein isothiocyanate-conjugated fibrinogen confirmed that binding of human fibrinogen to the conidia was dose dependent and specific. Binding was inhibited by unlabeled fibrinogen and by basement membrane laminin. Moreover, the expression of fibrinogen receptors at the surfaces of conidia seemed to be related to the maturation of the conidia. Binding sites appeared to be located in the D domains of the fibrinogen molecule. However, the peptide sequence recognized by the fungus could not be identified but was different from the classical adhesive recognition sequences, RGDS and fibrinogen gamma-chain dodecapeptide. In addition, an assay of adherence to proteins immobilized onto microtiter plates allowed us to establish the role of these interactions in fungal adhesion. Conidia strongly adhered to human fibrinogen and to laminin but not to fibronectin. Adhesion to fibrinogen substrates was specific, since it was inhibited by soluble fibrinogen and by specific antibodies, and seemed to be mediated by the D domains of the molecule. Study of the adhesion of numerous strains or clinical isolates to various mammalian fibrinogens did not reveal any particular affinity of strains for some animal species. Finally, by cultivation of the fungus in the presence of 125I-human fibrinogen and analysis of the radiolabeled material bound to the surface of the fungus, we were able to specify the sequence of events allowing its installation within the host. The interactions identified here may play an important role in governing fungal adherence and host tissue invasion.
During the last few years, several works have demonstrated the fixation of different host proteins on Aspergillus fumigatus conidia. Thus, after incubation in the presence of normal human plasma, the C3 component of complement is detected at the surface of conidia. In fact, most of the C3 deposited on conidia is converted in C3b or iC3b which would facilitate their phagocytosis by the macrophages. In the non immune host, the activation of the alternative pathway seems to be the main mechanism of the activation of the complement system by the conidia, but the participation of the classical pathway initiated by the fixation of the C-reactive protein has also been suggested. Aspergillus fumigatus conidia interact also with fibrinogen and laminin. These interactions which are mediated by the D domains of fibrinogen and by the fragment P1 of laminin, are specific. The number of fibrinogen binding sites at the surface of conidia has been calculated to be 1200 by cell, and the dissociation constant 2.2 x 10(-7) M. These interactions could determine the adhesion of conidia to the host tissues as suggested by adherence assays of conidia to proteins immobilized onto wells of microtiter plates. Conidia would bind to the fibrin deposits formed on damaged epithelia in response to the inflammatory reaction, or directly to laminin of the subepithelial basement membrane. Finally, different experiments suggested the identity of the binding sites for C3, fibrinogen and laminin at the surface of A. fumigatus conidia.
Aspergillus fumigatus, the causative agent of human aspergillosis, binds to and degrades basement membrane laminin. Using immunoelectron microscopy, laminin binding appeared to be associated with the cell wall expansions of resting conidia, and progressively extended to the outer electron dense layer of the conidial wall during the germination process. Labeling of thin sections revealed numerous binding sites in the cytoplasm, whereas the inner cell wall and the plasma membrane were not labeled. Attachment of A fumigatus conidia on microtiter plates coated with laminin and its fragments P1 and E8 was also investigated. Conidia cells showed good adhesion to wells coated with laminin. As indicated by inhibition experiments, the interaction was specific and fragment P1 represented the major binding site on the laminin molecule. In addition, since A fumigatus produced an extracellular serine protease, we determined the susceptibility of laminin to this enzyme. We demonstrated that protease extract was capable to degrade laminin in solution as well as in tissue sections. The laminin cleavage products were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All the three chains were extensively degraded within 1 h. Treatment of the crude protease extract with the enzyme inhibitors, phenylmethylsulfonyl-fluoride and chymostatin, blocked the degradation of laminin, indicating a chymotrypsin-like serine protease activity. Immunofluorescence microscopy of cryostat sections of mouse and rat kidneys treated with the protease extract showed widespread loss of laminin epitopes from basement membranes. Enzyme treatment also removed immunoreactivity from lungs as observed after immunoperoxidase performed on paraffin sections. Binding and proteolytic degradation of laminin may together facilitate initial interaction of A fumigatus with the host tissues.
To get a better understanding of the role of the previously reported fibrinogenolytic enzyme of Aspergillus fumigatus, we investigated the in vitro conditions of enzyme synthesis and attempted to characterize it. Modification of the nitrogen source did not influence the extracellular serine-proteinase profile, but resulted in important quantitative differences in the yields in batch cultures. The enzyme synthesis appeared to be an inducible phenomenon in A. fumigatus since it was initiated exclusively in the presence of proteins or protein hydrolysate. Free amino acids or inorganic nitrogen compounds could not promote significant enzyme production. Moreover, peptone at a concentration of 0.1% appeared to be the best inducer of enzyme synthesis. Conversely, modification of the carbon source did not affect fungal growth or enzyme synthesis. However, the production of chymotrypsin was highly sensitive to the carbohydrate level in the culture medium and, with peptone as nitrogen source, highest yields were obtained in the presence of 0.3 or 0.5% glucose. Culture filtrates of A. fumigatus CBS 113.26 grown with peptone or nitrate as nitrogen source were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparison of the protein patterns suggested for the proteinase a molecular mass of 33 kDa which was confirmed by chromatographic purification of the enzyme through (N alpha-CBZ)-D-phenylalanine agarose.
A fibrinogenolytic proteinase has been isolated from Aspergillus fumigatus culture filtrate by ammonium sulfate precipitation followed by successive chromatographies on Sephadex G-75 and immobilized phenylalanine. The purified proteinase exhibited a molecular weight of about 33 kDa. When analysed by SDS-polyacrylamide gels containing co-polymerized fibrinogen, the proteinase appeared as a broad band at the top of the gels, which could correspond to polymerization of the enzyme, as suggested by SDS-PAGE analysis of the unboiled eluate. The isoelectric point was 8.75 and the enzyme was not glycosylated. Proteinase activity was optimum at pH 9 and between 37 and 42 degrees C, although a decrease in activity was observed above 37 degrees C. PMSF and chymostatin markedly inhibited the proteinase activity, and good kinetic constants were obtained for the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA. These results provide direct evidence that this enzyme belongs to the chymotrypsin-like serine proteinase group.
The interaction of purified human fibrinogen with Aspergillus fumigatus conidia was investigated by immunofluorescence and electron microscopy and binding assays with radiolabeled proteins. We described the localization of the binding sites on the A. fumigatus conidia and on the fibrinogen molecule and determined the binding characteristics. Immunofluorescence revealed that the fixation of purified fibrinogen was selectively associated with conidia and suggested a role for the D domains of the fibrinogen molecule. Binding assays performed with 125I-radiolabeled proteins confirmed that binding sites were located specifically in the D domains. No reaction could be detected with fragment E. The binding of 125I-fragment D to conidia was time dependent, saturable, and specific. Scatchard analysis of the data revealed an average of 1,200 binding sites per conidium, and an apparent dissociation constant (Kd) of 2.2 x 10(-9) M was estimated. Pretreatment of the cells with proteolytic enzymes or heat abolished binding, demonstrating the protein nature of the binding sites. Ultrastructural localization of the fungal receptors was determined by transmission electron microscopy. Labeling appeared to be associated with the outer electron-dense layer of the conidial wall and progressively decreased during the germination process. Labeling of thin sections with fragment D and an antifibrinogen immune serum revealed that binding sites also lay in the inner part of the wall and in vacuoles. These results indicate the presence at the conidial surface of specific receptors for fibrinogen which could act as mediators of conidial adherence to host tissues.
Peritoneal dirofilariasis probably due to Dirofilaria repens is reported from a 52 years old woman. The parasite discovery is fortuitous during a surgical cure of intestinal adenocarcinoma. The patient lives in Provence and Corsica (Southern France), an endemic region. The authors present the histological aspects of the parasite.
Skin ulcer due to Mycobacterium ulcerans is presented. The patient come from East of French Guyana. Growth of this mycobacteria is obtained with diphasic Lowenstein medium at 30 degrees C. Diagnostic of M. ulcerans results from mycolic acids study.
Lungs can be the target of many fungal parasitic organisms. Some of them are specific to the lung (Pneumocystis); while others will migrate there in the course of their life cycle (Ascaris) or be caught in it (Paragonimus, Echinococcus). Sometimes, they also generate hypersensitivity reactions (fungi). In making a diagnosis one has to consider the present state of health or the disease history of the patient. A great number of parasites or mycetes are opportunist agents found in immuno-compromised patients. The epidemiological investigation giving details of a trip, even a long time ago, in a tropical region allows the suspicion of an imported parasitosis or an exotic mycosis. At present, we have on one hand, the mixing of populations, and on the other hand, many immuno-compromised situations which has given rise to a renewed interest in fungal and parasitic lung diseases.
In order to appreciate the pathogenicity of several geophilic Chrysosporium species (including Anixiopsis stercoraria, Chrysosporium keratinophilum, C. tropicum, C. pannorum, C. state Arthroderma curreyi, C. state of A. multifidum, and C. state of A. tuberculatum), the authors have realized two series of experimental infestations. Inoculation of these fungi on the back of guinea pigs produced rare erythematous scaling lesions which spontaneously disappeared 3-5 weeks later. No real hair invasion was observed. In white mice, eight weeks after intraperitoneal inoculation, granulomas with necrotic center were observed in the peritoneal tissue with C. keratinophilum, C. tropicum, C. state of A. curreyi and C. state A. tuberculatum. Conidia were often intact in necrotic centers and retrocultures were positive. With C. state of A. curreyi, spherical spores associated with rare budding cells were noted. The pathogenic role of these keratinophilic fungi is uncertain. However, their ability to remain viable for several weeks in skin and peritoneal tissue indicates that they could become pathogen in certain circumstances.
We report a case of congenital malaria due to a chloroquine-resistant strain of Plasmodium falciparum acquired in Mali. Ours is the first report of chloroquine-resistant congenital malaria in this part of Africa. We recall the various pathophysiologic, diagnostic and therapeutic features of this disease that should be considered in every neonate born to a mother who may have malaria. Although such cases are infrequent, we also discuss the very serious problems, mainly therapeutic, that they raise in several countries where they are endemic (South-East Asia and Africa particularly).