Biomedical subjects

I M Cesari

Publications and source records attributed to I M Cesari.

40 records · Page 3Linked to original sources

PubMedDate not supplied

Specificity of the solid phase alkaline phosphatase immunocapture assay for the diagnosis of human Schistosoma mansoni infection.

The species specificity of the solid phase alkaline phosphatase immunocapture assay (APIA) for the immunological detection of human immunoglobulin G antibodies to the alkaline phosphatase of adult Schistosoma mansoni was evaluated. Sera from schistosomiasis patients from South America, West Africa, south-east Asia and uninfected control subjects were compared. Only the sera of patients infected with S. mansoni gave positive results. There was no apparent difference between 2 populations infected with S. mansoni, one from South America and the other from West Africa. The results with sera from various regions of West Africa were also indistinguishable. Although the APIA was not able to discriminate the geographical origin of the S. mansoni-infected subjects, the method appeared to be specific for S. mansoni and suitable for use in the immunodiagnosis of schistosomiasis mansoni, particularly in endemic areas where mixed infections of Schistosoma spp. occur.

Alkaline Phosphatase↗

PubMedDate not supplied

Tegumental Ca-stimulated adenosine triphosphatase activity in adult Schistosoma mansoni worms.

A Ca-stimulated ATPase activity (pH 9.5) associated with the tegumental membrane enriched (TME) fraction of Schistosoma mansoni adults was partially inhibited by NAP-taurine or by increasing concentrations of chlorpromazine; endogenous calmodulin was found associated with the TME fraction. A similar activity (pH 8.6) was histochemically visualized within the tegument of fixed worms on the cytoplasmic leaflet of both the double surface membrane and the basement membrane; this reaction was inhibited by 1 microM chlorpromazine and it was also observed on the inner side of double membrane vesicles present in the TME fraction. No ATPase activity could be seen at alkaline pH with added Mg or Na/K ions. Without ATP, the addition of external Ca to the fixed worms induced the appearance of lead precipitates on the tegumental discoid bodies; this reaction was inhibited by molybdate and not by chlorpromazine. The intrategumentary regulation of calcium by the systems described and the possible use of phenothiazines against schistosomes are discussed.

Animals↗

PubMedDate not supplied

Immunodiagnosis of Schistosomiasis mansoni with APIA (alkaline phosphatase immunoassay).

The previously shown antigenicity of Schistosoma mansoni (JL venezuelan strain) alkaline phosphatase (Mg2+, pH 9.5) allowed its use in an immunodiagnosis assay, that consisted in the immunoadsorption of the enzymatic activity. Protein-A coated polyvinyl plates were used as solid phase to capture IgG from sera of infected human patients. After buffered saline washings, the plates were incubated with an enzyme-rich fraction (a n-butanol aqueous extract of adult worm obtained from pairs). Immunoadsorbed alkaline phosphatase (AP) was revealed by adding rho-nitrophenyl phosphate. Anti-AP antibodies were detected in 93% of coproparasitologically proven S. mansoni-infected venezuelan patients but not in parasite-free control sera and sera from patients infected with parasitosis other than schistosomiasis. The APIA did not correlate with cure but the anti-AP antibody response was progressively reduced after treatment. The use of an AP substrate amplifying system allowed an improvement of the assay sensitivity without loss of specificity. The data suggest that the APIA could be used as a marker of infection by S. mansoni.

Alkaline Phosphatase↗

PubMedDate not supplied

Schistosoma mansoni: surface membrane isolation with lectin-coated beads.

Lectins from Lens culinaris and Arachis hypogaea immobilized on polyacrylamide beads were used for selective isolation of glycosylated surface membrane domains of adult Schistosoma mansoni worms, and the method was compared with the membrane isolation procedure developed with polycationic (Affi-Gel) beads. The lentil lectin proved to be suitable for interaction with surface membrane components: an increment in the specific activities of tegumental phosphohydrolases was observed in the bound fraction with respect to that observed in a total worm homogenate. A characteristic polypeptide pattern on gel electrophoresis was also seen, more restricted than that obtained with the bound Affi-Gel fraction. Immobilized peanut lectin was not successful as a method for isolating membrane material from the tegument of adult worms. Solubilization and dissociation of the lentil lectin-bound enzyme markers was achieved after addition of detergent and competing sugars. Glycosylation of the solubilized enzymes was further confirmed by affinity chromatography with fresh lentil lectin-coated beads. These results, together with histochemical evidences, suggest that the active sites of some of these enzymes are located within or close to the cytoplasmic leaflet of the surface tegumental membranes, and allow us to propose a model for the double surface membrane complex where some proteins may be crossing the two bilayers.

Animals↗