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Biomedical subjects

D S Rosa

Publications and source records attributed to D S Rosa.

6 recordsLinked to original sources

Reservosome: an endocytic compartment in epimastigote forms of the protozoan Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). Correlation between endocytosis of nutrients and cell differentiation.

Reservosomes are large membrane-bound organelles found at the posterior end of epimastigote forms of Trypanosoma cruzi, but absent in amastigotes and trypomastigotes. We have transferred bloodstream trypomastigotes to LIT medium supplemented with gold-labelled transferrin in order to analyse, at the ultrastructural level, the occurrence of reservosomes and endocytosis during the trypomastigote to epimastigote differentiation. After 24 h, the trypomastigotes differentiated into amastigotes, which adhered to each other forming large clusters. Electron-dense vesicles were detected close to the Golgi complex in cells with intermediary characteristics between amastigotes and epimastigotes, but typical reservosomes at the posterior cell tip were still absent. Transferrin-gold complexes were observed only bound to the surface of clustered cells. After 72 h, epimastigotes were observed being released from the clusters and free-swimming epimastigotes appeared, containing electron-dense vesicles at their posterior region. Typical reservosomes, labelled with transferrin-gold, were observed only in free-swimming epimastigotes. When fully differentiated epimastigotes were incubated with transferrin-gold complexes and then processed for the immunocytochemical detection of cysteine proteinase, all reservosomes were positive for the enzyme, but co-localization of both markers did not occur in all organelles. Our data demonstrate that in T. cruzi epimastigotes endocytosis is strongly related to reservosome biogenesis during the trypomastigote to epimastigote differentiation process.

Animals↗

Excretory-secretory antigens of Trypanosoma cruzi are potentially useful for serodiagnosis of chronic Chagas' disease.

The reactivities of sera from chronic chagasic patients against the trypomastigote excreted-secreted antigens (TESA) of Trypanosoma cruzi strains with different biodemes were analyzed by TESA-blot and TESA-enzyme-linked immunosorbent assay (ELISA). Although both tests presented high sensitivity and specificity, TESA-ELISA is more appropriate for screening a larger number of samples.

Animals↗

Serodiagnosis of chronic Chagas infection by using EIE-Recombinant-Chagas-Biomanguinhos kit.

A kit based on an enzyme immunoassay, EIE-Recombinant-Chagas-Biomanguinhos, developed by the Oswaldo Cruz Foundation, was evaluated for the serodiagnosis of chronic Chagas disease. Evaluation was performed with 368 serum samples collected from individuals living in an endemic area for Chagas disease: 131 patients in the chronic phase with confirmed clinical, epidemiological, and serological diagnosis (indirect immunofluorescence, indirect hemagglutination or enzyme-linked immunosorbent assay) and 237 nonchagasic seronegative individuals were considered negative control. The EIE-Recombinant-Chagas-Biomanguinhos kit showed high sensitivity, 100% (CI 95%: 96.4-100%) and high specificity, 100% (CI 95%: 98-100%). The data obtained were in full agreement with clinical and conventional serology data. In addition, no cross-reaction was observed with sera from patients with cutaneous (n=14) and visceral (n=3) leishmaniasis. However, when these sera were tested by conventional serological assays for Chagas disease, cross-reactions were detected in 14.3% and 33.3% of the patients with cutaneous and visceral leishmaniasis, respectively. No cross-reactions were observed when sera from nonchagasic seronegative patients bearing other infectious disease (syphilis, n=8; HTLV, n=8; HCV, n=7 and HBV, n=12) were tested. In addition, sera of patients with inconclusive results for Chagas disease by conventional serology showed results in agreement with clinical evaluation, when tested by the kit. These results are relevant and indicate that the referred kit provides a safe immunodiagnosis of Chagas disease and could be used in blood bank screening.

Adolescent↗

Differentiation of Trypanosoma cruzi epimastigotes: metacyclogenesis and adhesion to substrate are triggered by nutritional stress.

Differentiation of Trypanosoma cruzi epimastigotes to metacyclic trypomastigotes occurs in the insect rectum, after adhesion of the epimastigotes to the intestinal wall. We investigated the effect of the nutritional stress on the metacyclogenesis process in vitro by incubating epimastigotes in the chemically defined TAU3AAG medium supplemented with different nutrients. Addition of fetal bovine serum induced epimastigote growth but inhibited metacyclogenesis. In this medium, few parasites attached to the substrate. Ultrastructural analysis demonstrated reservosomes at the posterior end of the epimastigotes. Incubation of the cells in TAU3AAG medium containing gold-labeled transferrin resulted in high endocytosis of the marker by both adhered and free-swimming epimastigotes. No intracellular gold particles could be detected in trypomastigotes. Addition of transferrin gold complexes to adhered epimastigotes cultivated for 4 days in TAU3AAG medium resulted in decrease of both metacyclogenesis and adhesion to the substrate, as compared with parasites maintained in transferrin-free medium. Adhesion to the substrate is triggered by nutritional stress, and proteins accumulated in reservosomes are used as energy source during the differentiation. A close relationship exists among nutritional stress, endocytosis of nutrients, adhesion to the substrate, and cell differentiation in T. cruzi epimastigotes.

Animals↗

Wound healing following excimer laser radial keratotomy.

A pulsed excimer laser was used to produce radial keratotomy in vivo in human corneas and the wound healing process was studied. Fluences of 370 mJ/cm2 at a repetition rate of 20 Hz were effective and atraumatic. The wound healing process was remarkably uneventful without an inflammatory or immune reaction apparent at 21 days. Consistent flattening of the cornea was obtained by shallower cuts than with conventional radial keratotomy techniques.

Adult↗

Laser in situ keratomileusis for hyperopia.

OBJECTIVE: To evaluate laser in situ keratomileusis (LASIK) ablation zone size and its relationship to refractive and visual outcomes in the treatment of hyperopia. METHODS: We evaluated retrospectively 40 eyes of 20 patients who had LASIK: 20 eyes had a refractive ablation zone ranging from 5.5 to 9.0 mm (Group 1); Group 2 consisted of 20 eyes with a refractive ablation zone from 5.5 to 8.25 mm. We used the Nidek EC-5000 excimer laser and the Hansatome microkeratome. Mean spherical equivalent refraction was +2.72 D in Group 1 and +2.75 D in Group 2. Objective refraction and visual acuity were evaluated over 6 months in both groups. RESULTS: Six months after LASIK, mean spherical equivalent refraction in Group 1 was +1.00 +/- 0.84 D and in Group 2 it was +0.75 +/- 0.42 D. One patient in Group 1 lost 1 line of spectacle-corrected visual acuity; none in Group 2 lost lines. Six patients gained 1 to 3 lines from their preoperative spectacle-corrected visual acuity. CONCLUSIONS: LASIK provides good results for mild and moderate hyperopia. LASIK with an ablation zone from 5.5 to 8.25 mm showed better predictability and more stable results. However, corneal diameter and the thickness and width of the flap appear to be important factors in the feasibility of LASIK in hyperopic eyes.

Adult↗