Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis

The antigenicity of promastigotes of Leishmania braziliensis braziliensis (L. b.braziliensis) treated with 1% sodium desoxycholate in 10 mM Tris-Hcl pH 8.2 was analysed by immunoblot using as probes sera from American cutaneous leishmaniasis (ACL), American visceral leishmaniasis (AVL), schistosomiasis, malaria and Chagas' disease. The ACL sera reacted constantly with a 60 kD band. No reactivity to this protein was observed with sera from the other diseases above mentioned indicating that the 60 kD protein may be used in serodiagnosis for ACL.

The sensitivity, specificity and efficiency values of some serological tests used in the diagnosis of paracoccidioidomycosis

This work reports on the results of double immunodiffusion (ID), counterimmunoelectrophoresis (CIE), complement fixation (CF) and indirect immunofluorescence (IIF) techniques in the serodiagnosis of paracoccidioidomycosis. The study was undertaken on four groups of individuals: 46 patients with untreated paracoccidioidomycosis, 22 patients with other deep mycoses, 30 with other infectious diseases (tuberculosis and cutaneous leishmaniasis) and 47 blood donors as negative controls. Data were obtained using Paracoccidioides brasiliensis antigens, i.e.,a yeast culture filtrate for ID, CIE and CF, and a yeast cell suspension for IIF. The sensitivity, specificity and efficiency values were measured according to GALEN & GAMBINO8.The gel precipitation tests (ID and CIE) showed the greatest sensitivity (91.3 and 95.6%, respectively), maximum specificity (100%) and the highest efficiency values when compared to the CF and IIF tests.

Self-diagnosis of active head lice infestation by individuals from an impoverished community: high sensitivity and specificity

To compare sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of self-diagnosis for head lice infestation with visual inspection, we conducted a study in an urban slum in Brazil. Individuals were asked about active head lice infestation (self-diagnosis); we performed visual inspection and thereafter wet combing (gold standard). Of the 175 individuals included, 77 (44%) had an active head lice infestation. For self-diagnosis, sensitivity (80.5%), specificity (91.8%), PPV (88.6%) and NPV (85.7%) were high. Sensitivity of visual inspection was 35.1%. Public health professionals can use self-diagnosis as a diagnostic tool, to estimate accurately prevalence of pediculosis in a community, and to monitor ongoing intervention strategies.

APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

On occurrence, habitat specificity and natural history of adult tiger beetles (Coleoptera: Carabidae: Cicindelinae) near Manaus, Central Amazonia, and key to the larvae of tiger beetle genera

Over a seven year period from 1991 to 1997, 22 species of tiger beetles, representing nine genera, were recorded near Manaus, Brazil. In the Whitewaterfloodplains along the Rio Solimões-Amazonas (Ilha de Marchantaria), three diurnal species inhabit inundation forests and six species (two diurnal, four nocturnal) live in open areas. Data on their natural history and adaptation to living conditions in floodplains are presented. Fifteen species were located on non-flooded uplands (Reserva Florestal A. Ducke). Five diurnal species inhabit the forest floor, two species are canopy dwellers, and eight species (seven diurnal, one nocturnal) live in open areas on whitesand or laterite. Only one species, Pentacomia lacordairei, was found in both floodplain and upland forests. A key to the larvae of tiger beetle genera located near Manaus is presented.

The relation of the concentration of macronutrients in the substrate and in the foliage to cell wall thickness and cellulose concentration in the xylem of slash pine (Pinus elliotti)

Sand culture experiments, using a sub-irrigation technique, were installed in order to find out the effects of the macronutrients N, P, K, Ca, Mg and S on growth, aspect, mineral composition, length of fibers, thickness of cell wall and cellulose concentration in slash pine. The aim was to obtain, under controlled conditions, basic information which could eventually lead to practical means designed to increase the rate of growth and to make of slash pine a richer source of cellulose. Nitrogen, Phosphorus, Potassium Experiment A 3 x 3 x 3 factorial design with two replicates was used. Nitrogen was supplied initially at the levels of 25, 50 and 100 ppm; phosphorus was given at the rates of 5, 10 and 20 ppm; potassium was supplied at the rates of 25, 50 and 100 ppm; six months after the experiment was started the first level for each element was dropped to zero. Others macro and all micronutrients were supplied at uniform rates. Fifteen hours of illumination per day were provided. The experimental technique for growing the slash pine seedlings proved quite satisfactory. Symptoms of deficiency of nitrogen, phosphorus and potassium were observed, described and recorded in photographs and water colors. These informations will help to identify abnormalities which may appear under field conditions. Chemical analysis of the several plant parts, on the other hand, give a valuable means to assess the nutritional status of slash pine, thus confirming when needed, the visual diagnosis. The correctness of manurial pratices, on the other hand, can be judged with the help of the analytical data tabulated. Under the experimental conditions nitrogen caused the highest increases on growth, as measured by increments in height and dry weights, whereas the effects of phosphorus and potassium were less marked. Cellulose concentration was not significantly affected by the treatments used. Higher levels of N seemed to decrease both length of fiber elements and the thickness of cell wall. The effects of P and K were not well defined. Calcium, Magnesium, Sulfur Experiment A 3 x 3 x 3 factorial design with two replicates was used. Calcium was supplied initially at the levels of 12.5, 25 and 50 ppm; magnesium and sulfur were given at the rates of 6, 12.5 and 25 ppm. Other macro and micronutrients were supplied at uniform rates, common to all treatments. Three months after starting the experiment the first level for each element was dropped to zero. Symptoms of deficiency of calcium, magnesium and sulfur were observed, described and recorded as in the case of the previous experiment. Chemical analysis were made, both for mineral content and cellulose concentration. Length of fibers and thickness of cell wall were measured. Both calcium and magnesium increase height, sulfur failing to give significant response. Dry weight was beneficially affected by calcium and sulfur. The levels of calcium, magnesium and sulfur in the needles associated with deficiency and maximum growth are comparable with those found in the literature. Cellulose concentration increased when the level of sulfur in the substrate was raised. The thickness of cell wall was negatively affected by the treatments; no effect was observed with regards to length of fibers.

Role of divalent cations, pH, cytoskeleton componentes and surface charge on the adhesion of Trichomonas vaginalis to a polystyrene substrate

The process of adhesion of three different strains of Trichomonas vaginalis to a polystyrene substrate was analysed. The process of adhesion was dependent on the time of incubation and the pH of the phosphate-buffered solution (PBS) in which the parasites were suspended. The highest indices of adhesion were observed after an incubation time of 60 min at pH 6.6. The adhesion index increased when the parasites were incubated in the presence of culture media or when Ca++ or Mg++ was added to the PBS solution, whereas cytochalasin B, trypsin or neuraminidase reduced adhesion. Incubation of the parasites in the presence of poly-L-lysine facilitated the process of adhesion. Incubation of the parasites or polystyrene beads in the presence of poly-L-lysine led to important changes in their surface charge.

The use of dacron plates for DOT enzyme-linked immunosorbent assay (DOT-ELISA)

Dacron (polyethylenetherephthalate) is proposed as a matrix for dot-ELISA procedures, as an alternative to nitrocellulose. Plates of dacron were partially hydrazinolyzed and hydrazide groups introduced were converted to azide groups. The derivative dacron-antigen was covalently linked on to the plates through these azide groups. The derivative dacron-antigen was exaustively washed according to CROOK and antigen was still fixed onto the plates. Protein F1A purified from Yersinia pestis was used as a model. Triration of sera from immunized and non immunized rabbits against this protein was carried out by employing the dot-ELISA method. No significant difference was observed using dacron-antigen and nitrocellulose-antigen preparations. However, both procedures showed to have a significant better performance in comparasion with the passive hemagglutination method. The specificity and reproductibility of the dot-ELISA assay using both preparations showed a similar behaviour. Nitrocellulose preparation was stable at 4ºC, 28ºC and -20ºC for 90 days, whereas the dacron-antigen derivative was stable only when stored at 4ºC. Dacron-antigen derivative could be re-used when the spot developing was proceeded using 4-chloro-1-naphtol as substrate.

Hemagglutinating and fusogenic activities of Newcastle disease virus: studies on receptor binding specificity and pH-induced conformational changes

Vaccinal and wild strains of Newcastle Disease virus (NDV) were analyzed for cell receptor binding and fusogenic biological properties associated with their HN (hemagglutinin-neuraminidase) and F (fusion protein) surface structures respectively. The evaluation of the biological activities of HN and F was carried out respectively by determination of hemagglutinating titers and hemolysis percentages, using erythrocytes from various animal origins at different pH values. Significant differences in hemagglutination titers for some strains of NDV were detected, when interacting with goose, sheep, guinea-pip and human "O" group erythrocytes at neutral pH. Diversity of hemolysis percentagens was observed between different NDV strains at acid pH. These analysis were developed to evaluate particular aspects of the actual influence of the receptor specifity and pH on the receptor binding and fusogenic processes of Newcastle Disease viruses.

The carbohydrate-binding specificity and molecular modelling of Canavalia maritima and Dioclea grandiflora lectins

The carbohydrate-binding specificity of lectins from the seeds of Canavalia maritima and Dioclea grandiflora was studied by hapten-inhibition of haemagglutination using various sugars and sugar derivatives as inhibitors, including N-acetylneuraminic acid and N-acetylmuramic acid. Despite some discrepancies, both lectins exhibited a very similar carbohydrate-binding specificity as previously reported for other lectins from Diocleinae (tribe Phaseoleae, sub-tribe Diocleinae). Accordingly, both lectins exhibited almost identical hydropathic profiles and their three-dimensional models built up from the atomic coordinates of ConA looked very similar. However, docking experiments of glucose and mannose in their monosaccharide-binding sites, by comparison with the ConA-mannose complex used as a model, revealed conformational changes in side chains of the amino acid residues involved in the binding of monosaccharides. These results fully agree with crystallographic data showing that binding of specific ligands to ConA requires conformational chances of its monosaccharide-binding site.

Host Specificity of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) with Comments on the Drop-off Rhythm

The parasitic specificity of larval, nymph and adult Amblyomma cajennense on six different host species: Oryctolagus cuniculus, Rattus norvegicus, Gallus gallus domesticus, Anas platyrhynchus, Coturnix coturnix and Streptopelia decorata is described. In terms of the numbers of larvae and nymphs recovered, O. cuniculus was the best host species. The modal day for drop-off of larvae and nymphs was day three for the mammal hosts, but variable in the birds. We conclude that adult A. cajennense have a strong degree of specificity due to the fact that the tick failed to complete its life cycle on any of the evaluated hosts. The immature stages, on the other hand, showed a low level of specificity, most especially in the larval stage, indicating the existence of secondary hosts which probably serve as dispersers in the wild. The results also indicated a variable drop-off rhythm for larvae and nymphs in two periods, diurnal (6-18 hr) and nocturnal (18-6 hr), which differed depending upon the host.