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Biomedical subjects

D Camus

Publications and source records attributed to D Camus.

At least 19 recordsLinked to original sources

[Blastocystis hominis: in search of a disease, a misunderstood organism].

Blastocystis hominis is a micro-organism which remains somewhat mysterious. Having defined its present position in the classification of Protozoa, the authors describe its ultrastructure and morphology. Its epidemiology and pathogenicity are discussed in the light of experimental studies and human clinical data, especially in AIDS patients. Metronidazole seems to be the most active drug against this organism, but extreme caution must be exerted when the possible pathogenic property of B. hominis is discussed.

Animals

Is Pneumocystis carinii a deep mycosis-like agent?

Pneumocystis carinii is a widespread eukaryotic microorganism found in the lungs of healthy mammals, including humans. It is able to proliferate extensively in the alveoli, becoming an important agent of severe pneumonitis in immunosuppressed hosts, especially in persons suffering from AIDS. The taxonomic position of P. carinii is uncertain. Typical cytoplasmic organelles of eukaryotic cells have been found and described in the parasite. Biochemical research is hindered by the lack of an efficient in vitro culture system. Results of comparative study of nucleic acid sequences suggest that Pneumocystis is a fungus. However, ultrastructural, biochemical and nucleic acid homology insights appear as clearly insufficient to class Pneumocystis. Pneumocystis infection might be acquired, as deep mycoses, from environmental sources through the respiratory tract. Thus, the hypothesis of an environmental stage of the parasite must be considered. Pneumocystis might be seen as a widespread pathogenic dimorphous fungus. As fungal agents, P. carinii is able to disseminate from the infected lung to other organs. However, deep mycoses and pneumocystosis induce different histopathological changes in the host. Furthermore, deep fungal infections, unlike pneumocystosis, cannot be transmitted from one infested host to another one. Beside these two aspects, pneumocystosis shares many features with deep mycoses. Research on the epidemiology of pneumocystosis is needed.

Animals

High osmotic pressure for Pneumocystis carinii London Resin White embedding enables fine immunocytochemistry studies: I. Golgi complex and cell-wall synthesis.

A method for embedding Pneumocystis carinii in hydrophilic resin (London Resin White) has been developed for immunocytochemistry studies. Using high osmotic pressure (about 850 mosmol) from fixation to embedding, this method improved the preservation of the fine structure as well as the antigenicity of rabbit- and SCID mouse-derived P. carinii. Cytochemistry studies were performed using colloidal gold-conjugated lectins (concanavalin A, glycine max, Ulex europaeus) that reacted with the cytoplasmic components (endoplasmic reticulum, Golgi vesicles). Colloidal gold-conjugated streptavidin was also tested and was found to be reactive with the parasite cell wall and cytoplasmic components, which precludes its indiscriminate use in P. carinii immunocytochemistry studies.

Animals

High osmotic pressure enables fine ultrastructural and cytochemical studies on Pneumocystis carinii. I. Epon embedding.

High osmotic pressure was used to preserve the ultrastructure of rabbit-, SCID mouse-, and rat-derived Pneumocystis carinii organisms from osmotic stress during fixation. Organelles and cytosol were well preserved within the tonicity range of 850-1,300 mosmol. Under these experimental conditions, we determined that the endoplasmic reticulum was well developed in all parasite stages and could observe the Golgi complex, autophagic vacuoles, dense bodies, type II endoplasmic saccules, and the recently described outer surface membrane, which was found in all parasite stages. The biological implications of these findings are discussed.

Animals

Intramolecular mapping of Plasmodium falciparum P126 proteolytic fragments by N-terminal amino acid sequencing.

Protein P126 (also called P140, P113, SERA, SERP1) is a major parasitophorous vacuole antigen of Plasmodium falciparum. This protein is processed upon merozoite release into 2 fragments of 73 kDa (P73) and 50 kDa (P50), which are found in the culture medium. P73 is composed of 2 polypeptides of 47 and 18 kDa linked by disulfide bridges. In the presence of leupeptin, an inhibitor of serine and cysteine proteases which inhibits merozoite release, a 56-kDa intermediate product (P56) is recovered in the culture medium instead of P50. In order to map these proteolytic fragments on the 126-kDa precursor, we purified them from Plasmodium falciparum culture medium by immunoadsorption, SDS-electrophoresis and Western blotting on PVDF membrane and determined the N termini of P126, P73 (P47 and P18), P50 and P56. Comparison of these sequences with the amino acid sequence deduced from the P126 gene allowed the mapping of the different fragments on the precursor. P47 was at the N-terminal and P18 at the C-terminal end of P126. P56 and P50 had the same N-termini and were located in the middle of P126. This latter result indicates that the proteolysis of P56-P50 occurs at the C-terminus of P56. The peptide bonds cleaved by leupeptin-insensitive activities are Glu-Thr and Gln-Asp; C-terminal sequencing of P50 will be needed to identify the leupeptin-sensitive cleavage site.

Amino Acid Sequence

Endogenous superoxide dismutase activity in two Babesia species.

Babesia hylomysci and B. divergens were studied for superoxide dismutase (SOD) activity by enzyme assay and isoelectric focusing (IEF). In the two Babesia species, parasite-associated SOD is cyanide-insensitive and inhibited by H2O2, indicating that iron is the cofactor metal. Measurements of SOD activity from purified parasites show that the SOD activity detected in Babesia is, for the main part, due to an endogenous enzyme.

Animals

Presence of human antibodies reacting with Candida albicans O-linked oligomannosides revealed by using an enzyme-linked immunosorbent assay and neoglycolipids.

In order to study the presence of antibodies directed against Candida albicans O-linked oligomannosides (oligomannosides O) in patient sera, we have developed an enzyme-linked immunosorbent assay (ELISA) involving neoglycolipids constructed with these residues (NGLO). Oligomannosides O released by mild alkaline degradation of the C. albicans cell wall phosphopeptidomannan (PPM) contained one to seven mannose residues, among which the quantitatively major components, mannobiose and mannotriose, were shown by 1H nuclear magnetic resonance to contain exclusively alpha (1-2) linkages. The pool of oligomannosides was converted to neoglycolipids by coupling them to 4-hexadecylaniline in an equimolar reaction checked by thin-layer chromatography. We have tested against these neoantigens, coated on ELISA plates, 15 pairs of sera corresponding to individual seroconversions observed in 15 patients during the course of a mycological and serological survey of candidiasis. For all patients, seroconversions resulted in an increased level of antibodies against NGLO. A significant correlation was observed between the results of ELISA-NGLO, ELISA involving the original PPM molecule, and routine antibody detection tests, indirect immunofluorescence assay, and cocounterimmunoelectrophoresis. These results therefore demonstrate the synthesis of human antibodies reactive with oligomannosides O constitutive of the C. albicans mannan molecule which have been previously described as exhibiting an inhibitory effect on human lymphocytic proliferation.

Antibodies, Fungal

Immunogenicity and antigenicity of the N-term repeat amino acid sequence of the Plasmodium falciparum P126 antigen.

The P126 protein, a parasitophorus vacuole antigen of Plasmodium falciparum has been shown to induce protective immunity in Saimiri and Aotus monkeys. In the present work we investigated its immunogenicity. Our results suggest that the N-term of P126 is poorly immunogenic and the antibody response against the P126 could be under a MHC restricted control in C57BL/6(H-2b) mice, which could be problematic in terms of a use of the P126 in a vaccine program. However, we observed that a synthetic peptide, copying the 6 octapeptide repeat corresponding to the N-term of the P126, induces an antibody response to the native molecule in C57BL/6 non-responder mice. Moreover, the vaccine-P126 recombinant induced antibodies against the N-term of the molecule in rabbits while the unprocessed P126 did not.

Amino Acid Sequence

[Prevention and treatment of materno-fetal toxoplasmosis].

Preventing congenital toxoplasmosis is fundamental, and in France screening tests for primary infection are obligatory for pregnant women. All nonimmunized women should be encouraged to follow good dietary and general health rules until delivery. Nevertheless, the risk of seroconversion does exist but can be detected early through monthly serum tests. Spiramycin as initial treatment of maternal primary infection is an essential step in preventing parasite transmission to the fetus. Fetal antitoxoplasmosis antibodies and indirect signs of congenital abnormalities should be detected early by amniotic fluid and fetal blood tests before echographic evidence confirms the diagnosis. Pyrimethamine-sulfonamide therapy should be undertaken and modulated according to laboratory results in order to prevent or treat any possible fetopathy. Congenital toxoplasmosis can be prevented in utero through biological diagnosis and specific therapy.

Adult

Characterization of microneme proteins of Toxoplasma gondii.

Three microneme proteins of Toxoplasma gondii have been characterized using 3 monoclonal antibodies and a recombinant protein specific antiserum. In all cases, apical labeling of tachyzoites and bradyzoites was observed by indirect immunofluorescence assay. Immunogold localization on ultrathin sections of bradyzoites or tachyzoites showed a specific labeling of micronemes. The following proteins were characterized using 2-dimensional gel electrophoresis and Western immunoblotting: Mic 1 (60 kDa, Pi 6.5), Mic 2 (120 kDa, Pi 5) and Mic 3 (90 kDa, Pi 6.75). The 90-kDa protein (Mic 3) is a heterodimer of two 38-kDa polypeptides (Pi 6.7 and 6.75 respectively) linked by disulfide bridges. Metabolic labeling and immunoprecipitation assays showed that at least one of the 38-kDa polypeptides was processed from a 40-kDa precursor. No processing was observed during the biosynthesis of the 120- and 60-kDa polypeptides.

Animals

Imaging of systemic Candida albicans infections with a radioiodinated monoclonal antibody: experimental study in the guinea pig.

Guinea pigs intravenously infected with Candida albicans were scanned to evaluate the use of radioiodinated monoclonal antibodies (MAb) to fungal antigens for detecting tissue infection sites. A total of 18 infected and 8 uninfected animals were used. MAb and F(ab')2 fragments directed against cell wall glycoproteins of C. albicans were labeled with 131I. Another MAb directed against a Schistosoma mansoni glycoprotein was labeled with 125I and used as a nonspecific control. Radiolabeled MAbs were injected at a dose of 12.5 micrograms (500 kBq) per animal. Images were acquired 24 h later. Animals were then killed and the dissected organs were separately gamma-counted. The number of C. albicans colony forming units (cfu) per gram was determined in each organ. A clear relationship was found between the anatomic distributions of C. albicans and 131I. The biodistribution of 131I radioactivity associated with anti-Candida MAb was greater in infected animals than in healthy animals and increased with the number of cfu per g in each organ. The distribution was highly specific in animals with Candida endophthalmitis, a pathognomic feature of organ involvement during hematogenous dissemination. In contrast, the distribution of 125I radioactivity associated with the nonspecific MAb was similar in healthy and infected animals. In infected animals, it was totally independent of the intensity of fungal infection.

Animals

Differential targeting of dense granule proteins in the parasitophorous vacuole of Toxoplasma gondii.

The biosynthesis and fate of 4 different dense granule proteins of Toxoplasma gondii were studied with 3 monoclonal antibodies raised against tachyzoites and 1 polyclonal antibody raised against a recombinant protein. These proteins have the following molecular weights: 27 kDa (GRA 1), 28 kDa (GRA 2), 30 kDa (GRA 3) and 40 kDa (GRA 4). All four proteins were found in dense granules by immunoelectron microscopy; in T. gondii-infected cells, they were found in the vacuolar network but, in addition, GRA 3 was also detected on the parasitophorous vacuole membrane. Therefore, dense granule contents undergo differential targeting when exocytosed in the parasitophorous vacuole. Metabolic labelling and immunoprecipitation showed that GRA 2 and GRA 3 were processed from lower molecular weight precursors, and that GRA 2 and GRA 4 incorporated [3H] glucosamine and are thus likely to be glycosylated.

Animals

Characterization of bradyzoite-specific antigens of Toxoplasma gondii.

Monoclonal antibodies that react specifically with bradyzoite antigens of Toxoplasma gondii were selected by differential immunofluorescence among hybridomas produced against these organisms. These antigens were further characterized by immunofluorescence on living bradyzoites and Western immunoblotting. Four pellicular antigens (36, 34, 21, and 18 kDa) were identified; three of these are exposed on the surface of the organism and accessible to either antibodies, trypsin cleavage, or both of these surface-probing procedures. These antigens were found on recent human isolates of T. gondii, as well as on laboratory strain bradyzoites obtained from either brain cysts or in vitro-grown parasites.

Animals

Morphological evaluation of Pneumocystis carinii after extraction from infected lung.

Changes in Pneumocystis carinii induced by the extraction of the parasite from rabbit lung have been investigated. Samples obtained using 4 extraction methods were evaluated by light and transmission electron microscopy. Light microscopic evaluation was insufficient to give a measure of the P. carinii viability or to detect parasitic cellular alterations. In contrast, ultrastructural evaluation provided information on host and P. carinii cell integrity, which is a critical condition for viability. None of the tested methods was ideal. How thoroughly and in what shape P. carinii need to be extracted from tissues will determine which extraction technique is of best use.

Animals

[Detection of early humoral response in experimental toxoplasmosis in mice].

Experimental infection of mice by Toxoplasma gondii is the best way for antigen production or direct diagnosis. Early serological response from sera and peritoneal exudate of mice infected by T. gondii has been studied. In this report the authors show that specific antibodies (IgM) are found in the serum at 24 hours after intraperitoneal inoculation. Serological methods were unable to detect free antibodies in the peritoneal exudate. Specific antibodies coating the parasite surface were detected by direct immunofluorescence as soon as 48 hours post infection. Biochemical treatment usually used for antigen production (trypsin, pepsin) were unable to destroy this antibodies which where no longer detected after 2 mercapto-ethanol treatment. Control experiments showed that, in these conditions, 2 mercapto-ethanol treatment had no effect on the reactivity of surface antigens with antibodies. Therefore, this results show that checking for specific antibodies absorbed at the surface of T. gondii is a prerequisite of the use of antigen produced in vivo.

Agglutination Tests

Golgi complex and lysosomes in rabbit derived Pneumocystis carinii.

The ultrastructural morphology of Pneumocystis carinii obtained from nonimmunosuppressed rabbit is described in details. Golgi complex and primary lysosomes of P carinii are described here for the first time. They are easily revealed by the zinc iodide-osmium tetroxide cytochemical reagent. Thiamine pyrophosphatase and beta-glycerophosphatase activities are found in the parasite but cytidine 5' monophosphatase activity is not observed. A weak thiamine pyrophosphatase activity is detected in Golgi vesicles. An endomembranous saccular structure, present from the intracystic body stage to the precystic stage, apparently plays the role of secondary lysosome. A second type of endomembranous saccular structure, only present in the well developed trophozoitic and precystic forms is also described. The presence of carbohydrates in the cell wall of the parasite was demonstrated by periodic acid-thiosemicarbazide-silver proteinate staining and lectin concanavalin A labeling. The development of Golgi vesicles preceded the transition from double-layered to three-layered parasite stages.

Animals

[Immunological changes during the immediate postoperative period (author's transl)].

Postoperative variations in the different parameters of humoral and cellular immunity were studied in 18 adult patients in general surgery. Levels of immunoglobulins G (igG) fell and those of immunoglobulins M (igM) increased significantly. Lymphocytic proliferation under the influence of PHA showed a significant fall. Neither polynuclear function nor total complement activity were modified. Patients in whom the postoperative course was complicated by sepsis differed from the others by a lower activity of the lymphocyte transformation test and by a higher level of igG and igM on the day prior to surgery. In addition, deficiency in the lymphoblastic transformation test increased during the postoperative period, there was a further fall in IgG and IgM increased less in these patients than in those in whom the postoperative course was uncomplicated.

Adult