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Biomedical subjects

C B Mello

Publications and source records attributed to C B Mello.

14 recordsLinked to original sources

Antiserum against perimicrovillar membranes and midgut tissue reduces the development of Trypanosoma cruzi in the insect vector, Rhodnius prolixus.

Antiserum raised against Rhodnius prolixus perimicrovillar membranes (PMM) and midgut tissue interfered with the midgut structural organization and reduced the development of Trypanosoma cruzi in the R. prolixus insect vector. SDS-PAGE and Western blot analyses confirmed the specific recognition of midgut proteins by the antibody. Feeding, mortality, molt, and oviposition of the insects were unaffected by feeding with the antiserum. However, the eclosion of the eggs were reduced from R. prolixus females treated with antiserum. Additionally, in vivo evaluation showed that after oral treatment with the antiserum, the intensity of infection with the Dm-28c clone of T. cruzi decreased in the digestive tract of fifth-instar nymphs and in the excretions of R. prolixus adults. These results suggest that the changes observed in the PMM organization in the posterior midgut of R. prolixus may not be important for triatomine survival but the antiserum acts as a transmission-reduction vaccine able to induce significant decreases in T. cruzi infection in the vector.

Animals↗

A combined method for evaluating radon and progeny in waters and its use at Guarani aquifer, São Paulo State, Brazil.

A combined method for evaluating radon (222Rn) and progeny (214Pb and 214Bi) in water was developed by using inexpensive alpha scintillation counting and gamma ray spectrometry through NaI(Tl) scintillation detectors. A groundwater sample collected at the Poços de Caldas alkaline massif in Brazil was submitted to the technique in order to assure its applicability by comparing the volumetric activities by different methods. Similar volumetric activity was determined for 214Pb and 214Bi in the sample analyzed that is compatible with the expected condition of radioactive equilibrium between these nuclides. The combined method was successfully used to analyze groundwater samples from Guarani aquifer in São Paulo State, Brazil, and the results of the measurements indicated that 214Pb and 214Bi provide useful information concerning the evaluation of the drinking water quality in terms of radiological aspects. This is because they are directly identified in the water samples, without the need of requiring the assumption of the establishment of the transient equilibrium condition with its parent 222Rn.

Bismuth↗

Effect of blood components, abdominal distension, and ecdysone therapy on the ultrastructural organization of posterior midgut epithelial cells and perimicrovillar membranes in Rhodnius prolixus.

The effects of blood components, nerve-cord severance, and ecdysone therapy on the posterior midgut epithelial cells of 5th-instar Rhodnius prolixus nymphs 10 days after feeding were analyzed by transmission electron microscopy. Cutting the nerve-cord of the blood-fed insects partially reduced the development of microvilli and perimicrovillar membranes (PMM), and produced large vacuoles and small electrondense granules; insects fed on Ringer's saline diet exhibited well developed microvilli and low PMM production; swolled rough endoplasmatic reticulum and electrondense granules; Ringer's saline meal with ecdysone led to PMM development, glycogen particles, and several mitochondria in the cytoplasm; epithelial cells of the insects fed on Ringer's saline meal whose nerve-cord was severed showed heterogeneously distributed microvilli with reduced PMM production and a great quantity of mitochondria and glycogen in the cytoplasm; well developed microvilli and PMM were observed in nerve-cord severed insects fed on Ringer's saline meal with ecdysone; Ringer's saline diet containing hemoglobin recovered the release of PMM; and insects fed on human plasma showed slightly reduced PMM production, although the addition of ecdysone in the plasma led to a normal midgut ultrastructural organization. We suggest that the full development of microvilli and PMM in the epithelial cells depends on the abdominal distension in addition to ingestion of hemoglobin, and the release of ecdysone.

Animals↗

Cross-cultural differences for three visual memory tasks in Brazilian children.

Norms for three visual memory tasks, including Corsi's block tapping test and the BEM 144 complex figures and visual recognition, were developed for neuropsychological assessment in Brazilian children. The tasks were measured in 127 children ages 7 to 10 years from rural and urban areas of the States of São Paulo and Minas Gerais. Analysis indicated age-related but not sex-related differences. A cross-cultural effect was observed in relation to copying and recall of Complex pictures. Different performances between rural and urban children were noted.

Age Factors↗

Trypanosoma cruzi I and Trypanosoma cruzi II: recognition of sugar structures by Arachis hypogaea (peanut agglutinin) lectin.

Epimastigote culture forms of different isolates of Trypanosoma cruzi from different mammal hosts, humans, and vectors were tested with FITC-conjugated peanut agglutinin lectin (PNA-FITC). The parasites maintained in axenic medium, liver infusion tryptose. were evaluated by flow cytometric analyses; whereas T. cruzi I (Tcl), which is associated with the sylvatic transmission cycle, was labeled in high percentages with PNA (88-99.2%), T. cruzi II (TcII) (parasites associated with domiciliar cycle) and T. cruzi, zymodeme 3 (Tc/Z3) (also associated with the sylvatic cycle) were labeled in low percentages (TcII, 0-26% and Tc/Z3, 0-12.6%). It was demonstrated that it is possible to differentiate the 2 main T. cruzi subpopulations, TcI and TcII, using Arachis hypogaea. These results also showed a higher variability in TcII in terms of PNA binding.

Animals↗

Role of superoxide and reactive nitrogen intermediates in Rhodnius prolixus (Reduviidae)/Trypanosoma rangeli interactions.

This study compares aspects of the superoxide, nitric oxide and prophenoloxidase pathways in Rhodnius prolixus hemolymph, measured in parallel, in response to Trypanosoma rangeli inoculation. Responses to two strains of T. rangeli, and two developmental forms, were studied, and the results obtained were correlated with the ability of the parasites to survive, multiply, and complete their life cycles in the hemolymph of the host. T. rangeli H14 strain parasites, which fail to complete their life cycle in Rhodnius by invading the salivary glands, stimulated high levels of superoxide and prophenoloxidase activity, which peaked 24 h after inoculation. Simultaneously, the concentration of hemolymph nitrites and nitrates increased, indicative of nitric oxide activity, but parasite numbers remained low. T. rangeli Choachi strain parasite inoculation also stimulated superoxide and prophenoloxidase activity, which, though significantly lower than the equivalent responses to the H14 strain, also peaked at 24 h. However, nitrate and nitrite levels in Choachi strain-inoculated hemolymph remained low, and this parasite strain multiplied rapidly, especially following peak superoxide activity, and eventually invaded the salivary glands for transmission to a vertebrate host. In both strains, short form epimastigotes stimulated greater superoxide and prophenoloxidase responses than long form epimastigotes. Injection of the NADPH oxidase inhibitor N-ethylmaleimide or the inducible nitric oxide synthase inhibitor S-methyl isothiourea sulfate caused significantly higher insect mortalities in groups of R. prolixus inoculated with either parasite strain compared with those of uninfected control insects. This indicates that both NADPH oxidase and nitric oxide synthase activity may be involved in the immune response of R. prolixus to infection by T. rangeli. Finally, Western blotting of R. prolixus hemocyte lysates revealed the presence of a protein immunologically related to the human NADPH oxidase complex, the initiator enzyme of the respiratory burst.

Animals↗

Studies on a haemolymph lectin isolated from Rhodnius prolixus and its interaction with Trypanosoma rangeli.

We demonstrated that in Rhodnius prolixus haemocyte monolayers, both Trypanosoma cruzi and Trypanosoma rangeli are capable of inducing haemocyte/parasite clump formation. We also purified, by one-step affinity chromatography, a haemolymph galactoside-binding lectin from R. prolixus which we believe could play an important role in the development of T. rangeli in the haemocoel of the insect vector. This lectin markedly enhanced the activation of clump formation by T. rangeli in R. prolixus haemocyte monolayers, with an increase in clump size and haemocyte aggregation. The haemolymph lectin also significantly affected the motilitity and survival of T. rangeli culture short forms, but not the long forms, when they were incubated in vitro. This molecule is also one of the few described in insects with agglutination activity independent of calcium ions. The partial N-terminal amino acid sequence of this lectin demonstrated similarity to a bacterial xylulose kinase and in preliminary experiments the purified haemolymph lectin phosphorylated a tyrosine kinase substrate in a dose-dependent manner. The possible role of this haemolymph lectin in the life cycle of T. rangeli is discussed.

Amino Acid Sequence↗

Influence of brain and azadirachtin on Trypanosoma cruzi development in the vector, Rhodnius prolixus.

Studies on the effects of decapitation, head transplantation, azadirachtin, and ecdysone therapy on the ultrastructural organization of the midgut of Rhodnius prolixus, a vector of the protozoan Trypanosoma cruzi, show a distinct effect on the organization of the epithelial cells. When insects are decapitated or treated with azadirachtin, the ultrastructural organiza tion of these compartments changed significantly and drastically blocked the development of T. cruzi infection. In converse experiments, head transplantation or oral therapy with ecdysone significantly re versed the T. cruzi infectivity and reestablished the organization of the stomach and intestine in decapitated or azadirachtin-treated insects. These results indicat that a brain factor, possibly the prothoracicotropic hormone which stimulates ecdysteroid production on the prothoracic glands, may act directly or indirectly on both the midgut cell organiza tion and the intestinal microenvironment, interfering in the trypanosome survival and infection of the vector R. prolixus.

Animals↗

Differential in vitro and in vivo behavior of three strains of Trypanosoma cruzi in the gut and hemolymph of Rhodnius prolixus.

A comparison was made of the agglutination and lysis of three strains of Trypanosoma cruzi in gut extracts and hemolymph of Rhodnius prolixus and the results obtained were correlated with the success or failure of the parasite strain to infect the digestive tube or to survive in the hemocel after inoculation. Both T. cruzi strains Dm28c and Cl urine 35 days after feeding with parasites. Concomitantly, both of these strains were agglutinated but not lysed by the crop extracts. In contrast, T. cruzi Y strain parasites rapidly disappeared from the gut and showed no agglutination, but some lysis, in the crop extract. Following inoculation into the hemocel, only the Cl strain survived at high levels and was also the only strain agglutinated significantly in the hemolymph. Both Dm28c and Y strains rapidly disappeared from the hemocel with the former parasite being removed more slowly than the latter, probably due to clearance by the cellular defenses. The rapid clearance of the Y strain was correlated with the presence of a high titer lysin in the hemolymph. Subsequent experiments using FITC-labeled lectins and FACS to probe the carbohydrates on the parasite surfaces showed significant differences between the three strains. Thus, only Dm28c was stained strongly by Arachis hypogea (PNA) lectin, indicating the presence of galactose/N-acetylgalactosamine residues, and Dm28c and Y strains by Phytolacca lectin for N-acetyl glucosamine moieties. Finally, the fact that, in contrast to Dm28c and Y, the Cl strain strongly interacted with Triticum vulgaris (WGA) but not with Phytolacca lectin may be due to the presence of N-acetyl neuraminic acid residues on these organisms. These surface carbohydrate differences may be correlated both to the behavior and agglutination variations between the three strains recorded in this work.

Agglutination Tests↗

Trypanosoma cruzi and erythrocyte agglutinins: a comparative study of occurrence and properties in the gut and hemolymph of Rhodnius prolixus.

The activity of agglutinins found in the gut tissues and hemolymph of Rhodnius prolixus was tested using rabbit erythrocytes, Trypanosoma cruzi, or Trypanosoma rangeli as test particles. In addition, investigations were made of the influence of parasitic infection and insect diet on the agglutination titers. A range of physicochemical tests and carbohydrate-binding studies were performed and inhibitors were subsequently found for both the crop agglutinin (p-nitrophenol-derived sugars) and the hemolymph agglutinin (galactose-type sugars). As a result, affinity chromatography was utilized for an attempted purification of these agglutinins, and a one-step purification protocol for the R. prolixus hemolymph agglutinin has been developed. Preliminary results of some biological and physicochemical characteristics of this pure agglutinin are described. These results represent a starting point for future studies of lectin/parasite interaction in this reduviid-trypanosome model.

Animals↗

Trypanosoma cruzi and Trypanosoma rangeli: interplay with hemolymph components of Rhodnius prolixus.

Studies were carried out on the course of infection of Trypanosoma cruzi (clone Dm28c) and Trypanosoma rangeli (clone San Agustin) and their interactions with hemolymph components of Rhodnius prolixus. These parasites when inoculated into the hemocoel of adult R. prolixus (i) had different courses of infection (T. rangeli had high rates of both multiplication and infection and T. cruzi had no division and disappeared soon from the hemolymph); (ii) induced high but no differential increases in lysozyme levels; (iii) failed to induce any other antibacterial activity; (iv) showed similar patterns of hemolymph agglutination activity for erythrocytes and parasites, although there was evidence of limited, unquantifiable, agglutination of T. cruzi; (v) elicited different hemocyte responses with only the T. rangeli infection resulting in significantly increased hemocyte counts; and (vi) did not induce trypanolytic activity. These experiments, unlike previous studies, also showed (i) an interaction of these trypanosomes with the prophenoloxidase-activating system [phenoloxidase (PO) production was spontaneously activated by both parasites but the number of T. cruzi in the hemolymph was directly correlated with PO levels] and (ii) that the elimination of T. cruzi also corresponded to the formation of nodules in the hemolymph. The significance of these results is discussed in relation to the hypothesis that T. rangeli but not T. cruzi has the ability to escape from and perhaps utilize the vector immune system in order to successfully colonize the R. prolixus hemolymph.

Animals↗

Rhodnius prolixus: salivary antihemostatic components decrease with Trypanosoma rangeli infection.

Feeding behavior and some antihemostatic properties of the saliva of Rhodnius prolixus infected with Trypanosoma rangeli were studied and compared with those of uninfected controls. Insects with salivary gland infection pierced host skin more often and drew less blood and at a lower rate than controls when feeding on a rabbit. None of these differences was observed when feeding was performed through a membrane feeder. Salivary gland homogenates from infected insects, at 30 days after feeding/infection, had a significantly lower amount of total protein/salivary gland pair and less anticoagulant activity than controls. Also, infected salivary glands exhibited significantly reduced apyrase activities and reactive nitrogen groups when compared to controls. It is concluded that salivary infection of T. rangeli impairs the ability of the vector to locate blood vessels by affecting salivary antihemostatic properties, thus enhancing the possibility of intradermal inoculation of parasites into the mammalian host.

Animals↗

In vitro cytotoxicity of Rhodnius prolixus hemolytic factor and mellitin towards different trypanosomatids.

Trypanosoma cruzi strain Y and clone Dm28c and other trypanosomatids were exposed to two lytic agents, Rhodnius prolixus hemolytic factor (RHF) and mellitin, in vitro. In both cases, the result was a significant decrease in the number of parasites after a 30-min treatment at 37 degrees C. RHF and mellitin had distinct activities on different strains and species of trypanosomatids. These observations suggest that RHF may be an important factor in selecting resistant strains of trypanosomes for development in the vector's gut.

Animals↗

In vivo and in vitro metacyclogenesis tests of two strains of Trypanosoma cruzi in the triatomine vectors Triatoma pseudomaculata and Rhodnius neglectus: short/long-term and comparative study.

Metacyclogenesis of Trypanosoma cruzi of the Y and Berenice strains was studied in Triatoma pseudomaculata and Rhodnius neglectus. Results in vivo showed a higher production of metacyclic trypomastigotes in R. neglectus' digestive tube than in T. pseudomaculata. In vitro experiments were also carried out in order to compare the behavior of culture forms of T. cruzi incubated in extracts of different compartments (stomach, intestine, and rectum) of the digestive tract of both species of triatomines. A higher percentage of metacyclic trypomastigotes for both parasite strains, Y and Berenice, was detected in the rectum extract of R. neglectus in comparison to that from T. pseudomaculata. The same results were obtained with in vitro experiments, using parasites incubated in urine from each of those vectors. The adhesion of parasites to the incubated rectum epithelial cells was also compared. In incubations with the Y strain no significant differences were detected between the two triatomine species but, however, with the Berenice strain the mean percentage of cells with adhered parasites was higher in R. neglectus than in T. pseudomaculata.

Animals↗