Alternative approaches in schistosomiasis control

Measures for for the control of schistosomiasis were implemented in Egypt begining 1922. This shows that developing endemic countries are facing this problem for near 70 years. However, results in the control of this infection have not been satisfactorily obtained in spite of the technologies and strategies recently developed. The idea that the social and economic components are relevant in the control of schistosomiasis is not new although its extension and profundity have not usually been well understood. More recently; most of the workers have recognized that the distribution of the prevalence rates of schistosomiasis should not be neglected in the control of the infection. At present field work projects on the control of schistosomiasis are being developed in rural areas of two Brazilian states (Espirito Santo and Pernanbuco). The adopted strategy aims to interfere in the complex relationships between amn and his bio-social-cultural environment, without forgeting tha the unequal distribution of the space is a consequence of the political and economic organization of the Society.

Is the thymus a target organ in infectious diseases?

The thymus is a central lymphoid organ, in wich T cell precursors differentiale and generate most of the so-called T cell reprtoire. Along with a variety of acute infectious diseases, we and others determined important changes in both microenvironmental and lymphoid compartments of the organ. For example, one major and common feature observed in acute viral, bacterial and parasitic diseases, is a depletion of cortical thymocytes, mostly those bearing the CD4-CD8 double positive phenotype. This occurs simmultaneously to the relative enrichment in medullary CD4 or CD8 single positive cells, expressing high densities of the CD3 complex. Additionally we noticed a variety of changes in the thymic microenvironment (and particularly is epithelial component), comprising abnormal location of thymic epithelial cell subsets as well has a denser Ia-bearing cellular network. Moreover, the extracellular matrix network was altered with an intralobular increase of basement membrane proteins that positively correlated with the degree of thymocyte death. Lastly, anti-thymic cell antibodies were detected in both human and animal models of infectious diseases, and in some of them a phenomenon of molecular mimicry could be evidenced. Taken together, the data receiwed herein clearly show that the thymus should be regarded as a target in infectious diseases.

Cytological and isoenzyme analysis of the Bucay and Quevedo cytotypes of the onchocerciasis vector Simulium exiguum (Diptera: Simuliidae) in Ecuador

Four cytotypes of Simulium exiguum occur in Ecuador, where this morphospecies is the primary vector of onchocerciasis. In this paper, we give the first full description of the banding pattern of the larval polytene chromosomes of the Quevedo cytotypes differ from the chromosomal standard sequence (of the Cayapa cytotype) by the fixed inversions IIL-5 and IIL-6. The Quevedo cytotype additionally differs from the standard and Bucay cytotypes by processing a differentiated X chromosome, wich is indicated by the inversion IIS-A. As the degree of reproductive isolation between the Bucay and Quevedo cytotypes has not yet been estabilished, they must be regarded as intraspecific variants of the same species. In fact, isoenzyme characterizations showed that the Bucay and Quevedo cytotypes are differentiated only to the extent expected of incipient species or geographical populations. Moreover, the sibiling species status previously given to the Bucay cytotype needs be reassessed, there being inadequate analysis from areas in Ecuador where Bucay occurs in sympatry with the standard Cayapa cytotype. No isoenzyme electromorphs were discovered that identified all or mostadult females of any one (cytotype-pure) collection.

Description of Leishmania (Leishmania) forattinii sp. n., a new parasite infecting opossums and rodents in Brazil

A new parasite species of Leishmania is described, L. (Leishmania) forattinii sp. n., which was isolated from a pooled triturate of liver and spleen of a opossum (Didelphis marsupialis aurita) and from skin samples from a rodent (Proechmys iheringi denigratus), captured in primary forest on the Atlantic Cost of Brazil. Our results on the basis of biological and molecular criteria indicate that this taxonomically distinct parasite ias a new species of the L. mexicana complex, but closely related to L. (L.) aristidesi Laison & shaw, 1979, as revelated by phenetic and phylogenetic numerical analyses of the enzyme data. L. forattinii was clearly distinguishable from other Leishmania species of the genus usisng enzyme electrophoresis, monoclonal antibodies, molecular karyotypes, analysis of restriction enzyme digestion patterns of kinetoplast DNA (kDNA), as well as the use of kDNA hybridization procedures.

Elf-1 contributes to the function of the complex interleukin (IL)-2-responsive enhancer in the mouse IL-2 receptor alpha gene.

Lymphocytes regulate their responsiveness to IL-2 through the transcriptional control of the IL-2R alpha gene, which encodes a component of the high affinity IL-2 receptor. In the mouse IL-2R alpha gene this control is exerted via two regulatable elements, a promoter proximal region, and an IL-2-responsive enhancer (IL-2rE) 1.3 kb upstream. In vitro and in vivo functional analysis of the IL-2rE in the rodent thymic lymphoma-derived, CD4- CD8- cell line PC60 demonstrated that three separate elements, sites I, II, and III, were necessary for IL-2 responsiveness; these three sites demonstrate functional cooperation. Site III contains a consensus binding motif for members of the Ets family of transcription factors. Here we demonstrate that Elf-1, an Ets-like protein, binds to site III and participates in IL-2 responsiveness. In vitro site III forms a complex with a protein constitutively present in nuclear extracts from PC60 cells as well as from normal CD4- CD8- thymocytes. We have identified this molecule as Elf-1 according to a number of criteria. The complex possesses an identical electrophoretic mobility to that formed by recombinant Elf-1 protein and is super-shifted by anti-Elf-1 antibodies. Biotinylated IL-2rE probes precipitate Elf-1 from PC60 extracts provided site III is intact and both recombinant and PC60-derived proteins bind with the same relative affinities to different mutants of site III. In addition, by introducing mutations into the core of the site III Ets-like motif and comparing the corresponding effects on the in vitro binding of Elf-1 and the in vivo IL-2rE activity, we provide strong evidence that Elf-1 is directly involved in IL-2 responsiveness. The nature of the functional cooperativity observed between Elf-1 and the factors binding sites I and II remains unresolved; experiments presented here however suggest that this effect may not require direct interactions between the proteins binding these three elements.

Acetylated alpha-tubulin in Trypanosoma cruzi: immunocytochemical localization

We have used monoclonal antibodies specific for acetylated and non-acetylated alpha-tubulin to localize microtubules containing acetylated alpha-tubulin in all developmental forms of the life cycle of Trypanosoma cruzi. This was demonstrated using immunofluorescence and by transmission electron microscopy of thin sections, negative stained cells, and replicas of whole Triton X-100 extracted cells immunolabeled with antibody-gold complex. The antibody specific for acetylated alpha-tubulin (6-11B-1) binds to the flagellar, as well as to the sub-pellicular microtubules. The extent of labeling of the sub-pellicular microtubules with the monoclonal antibody recognized alpha-acetylated tubulin was smaller than that observed with the antibody which recognizes all tubulin isoforms. In relation to the developmental forms, the extent of labeling of the microtubules with antibody 6-11B-1 was larger in epimastigote and trypomastigote than in amastigote forms. Incubation of the parasites for 1 h at 0º C or in the presence of either colchicine or vinblastine did not interfere with the sub-pellicular microtubules. These observations, in agreement with those reported for Trypanosoma brucei brucei (Schneider et al., 1987; Schulze et al., 1987; Sasse per cent Gull, 1988) indicate that the sub-pellicular microtubules of trypanosomatids represent stable microtubules containing acetylated alpha-tubulin (or the alpha 3-tubulin isotype).

The Crocidura fuliginosa species complex (Mammalia, Insectivora) in Peninsular Malaya: biological, karyological and genetical evidence

Four West Malaysian shrew populations of the genus Crocidura were investigated through their karyotype and allozyme variations, and, in part, by interfertility experiments. Two different karyotypes characterize these shrews. The first, restricted to the Cameron Highlands (Peninsular Malaysia), invariably has 2n = 40 chromosomes but a varying fundamental number (FN = 54-58). The second karyotype shows a fundamental number of 62-68 and a polymorphic chromosomal number of 2n = 38, 39 or 40, a rare event in the genus Crocidura. Thus both can be distinguished by either a low or a higher number of meta- and submetacentric elements. In heterospecific breeding experiments, mutual avoidance was observed suggesting prezygotic barriers, whereas intraspecific pairs produced 13 liters (mean 2.1 young). Furthermore, our biochemical results indicate that both karyotypes correspond to a relatively ancient separation (Nei's D = 0.354), an amount of genetic differentiation comparable to the distance separating them from the West Palearctic C. russula (D = 0.429-0.583). In contrast, conspecific island and mainland Malaysian shrews possessing the second karyotype had only one fixed allelic difference over the 35 loci surveyed. The problem of naming the two biological species remains unsolved and requires further comparative investigations.

Protein recovery, separation and purification: selection of optimal techniques using an expert system

The paper discusses the utilization of new techniques ot select processes for protein recovery, separation and purification. It describesa rational approach that uses fundamental databases of proteins molecules to simplify the complex problem of choosing high resolution separation methods for multi component mixtures. It examines the role of modern computer techniques to help solving these questions.

Visceral leishmaniasis in Teresina, state of Piauí, Brazil: preliminary observations on the detection and transmissibility of canine and sandfly infections

A Leishmania donovani-complex specific DNA probe was usedto confirm the widespread dissemination of amastigotes in apparently normal skinof dogs with canine visceral leishmaniasis. When Lutzomyia longipalpis were fed on abnormal skin of five naturally infected dogs 57 of 163 (35 per cent) fliesbecame infected: four of 65 flies (6 per cent) became infected when fed on apparently normal skin. The bite of a single sandfly that had fed seven days previouslyon a naturally infected dog transmitted the infection to a young dog from a non-endemic area. Within 22 days a lesion had developed at the site of the infectivebite (inner ear): 98 days after infection organisms had not disseminated throughout the skin, bone marrow, spleen or liver and the animal was still serologically negative by indirect immunofluorescence and dot-enzyme-linked immunosorbent assay. When fed Lu. longipalpis were captured from a kennel with a sick dog known to be infected, 33 out of 49 (67 per cent) of flies contained promastigotes. In contrast only two infections were detected among more than 200 sandflies captured in houses. These observations confirm the ease of transmissibility of L.chagasi from dog to sandfly to dog in Teresina. It is likely that canine VL is the major source of human VL by the transmission route dog-sandfly-human. the Lmet2 DNA probe was a useful epidemiological tool for detecting L. chagasi in sandflies.

Gundlachia dutrae: n. sp. from northwest Brazil (Mollusca: Basommatophora: Ancylidae)

A new species of Gundlachia, Gundlachia dutrae is described from northwest Brazil. It is distinguishable from other congenerie species by characteristics of the shell, radula and internal organs. Shell relatively high. Aperture near-circular; periostracum dark brown without periostracal hairs. Apex slightly inclined to the right, projected but not hooked, with an apical depression surrounted by a sculpture of well-marked irregular punctations. Shell surface with prominent radial sculpture. No septate specimens were observed. Ratios (n= 59): shell width/shell lenght = 0,66- 0,79 (mean 0,73); shell height/shell length = 0,32- 0,45 ( mean 0,37); shell height/shell width = 0,43- 0,63 (mean 0,51). Body of normal ancylid type; mantle pigmentation dark brown or black, concentrated along the mantle collar. The dorsal surface of the right anterior muscle is elongated and medially constricted. The left anterior and the posterior muscles are almost elliptical. Adhesive area is V-shaped. Pseudobranch unpigmented bearing a very small and thin dorsal lobe. Ovotestis with more than 25 unbranched diverticula. Ovispermiduct with seminal vesicle rather developed. Elongated nidamental gland continous with the glandular wall of the uterus. Nidamental gland appendix ending into a bulbous swelling Spermathecal body almost rounded. Well-developed prostate with five long diverticula. Ejaculatory complex with long glandular flagellum, without a penis or true ultra-penis. "Penis sheath" developed. "Ultra-penis" projected as a tube inside the lumem of prepuce, with a slit between "ultra-penis" and "penis sheath". Rachidian tooth tetracuspid, with two median cusps assymmetrical and aculeated. Lateral teeth tricuspid, with a reduced endocon and a prominent mesocon. A well marked gap occurs between meso and ectocon. Marginal teeth similar to lateral ones. Jaw T-shaped, with about 28 dorsal plates.

Exploitation of parasite derived antigen in therapeutic success of human cutaneous leishmaniasis in Brazil

In a complete study in 25 patients with American cutaneous leishmaniasis, caused by Leishmania braziliensis complex, immunotherapeutic efficacy of parasite derived antigen (94-67 KD) has been compared to antimonial therapy. Additionally, to delineate the mechanism of therapeutic success, microscopical features of immune response in active lesions and healed or non-healed lesions following therapy were analyzed. The results showed that cure rates in immunotherapy and chemoterapy were equal (>83 por cento). The immunohistochemical changes in two therapeutic groups were also largely similar. The analysis of humoral and cellular immune response suggest that appropriate stimulation of T helper cells in the lesion site, in association with one or more cytokines, play a key role in the healing process.

Determination and control of schistosomiasis

The subject of this conference reflects the scientific community's interest in seeking to understand the complex causal web whose various social, economic, and biological components interact in the production and reproduction of schistosomiasis and its control in relation to community participation. From the onset, the author stresses the impossibility of dealing separately with community participation, as if social components were just one more "weapon" in the arsenal for schistosomiasis control. This study begins with a brief historical review of the 71 years of control activities with this endemic disease, stressing the enormous efforts and huge expenditures in this field vis-à-vis the limited results, despite the extraordinary technological development of specific, classical control inputs such as new treatment drugs and molluscicides. The article then discusses the various strategies used in control programs, emphasizing ideological consistencies and contradictions. Interactions at the macro and micro levels are discussed, as are the determinants and risk factors involved in producing the disease's endemicity. Unequal occupation of space leaves the segregated portion of the population exposed to extremely favorable conditions for transmission of the disease. This raises the issue of how to control an endemic disease which is so closely linked to the way of life imposed on the population. The study challenges the classical control model and suggests an alternative model now undergoing medium-term investigation in the States of Espirito Santo, and Pernambuco, Brazil. The author concludes that we do not need new strategies, but a new control model, contrary to the prevailing classical model in both concept and practice. From the conceptual point of view, the new model mentioned above is different from others in that schistosomiasis control is seen from a social perspective stressing the population's accumulated knowledge in addition to the building of shared knowledge. The model's praxis has the following characteristics: (1) it is integrated with and financed by research agencies and health services; (2) it operates at the local health services level; (3) use of molluscicides has been eliminated; (4) emphasis is given to individual medical treatment and improvement of sanitary conditions.

A molecular genetic study of the variations in metabolic function during schistosome development

During their complex life cycle schistosomes alternate between the use of stored glycogen and reliance on host glucose to provide for their energy needs. In addition, there is dramatic variation between the relative contribution of aerobic versus anaerobic glucose metabolism during development. We have cloned a set of representative cDNAs that encode proteins involved in glucose uptake, glycolysis, Kreb's cycle and oxidative phosphorylation. The different cDNAs were used as probes to examine the expression of glucose metabolism genes during the schistosome life cycle. Steady state mRNA levels from whole cercariae, isolated cercarial tails, schistosomula and adult worms were analysed on Northern blots and dot blots which were quantified using storage phosphor technology. These studies reveal: (1) Transcripts encoding glycogen metabolic enzymes are expressed to much higher levels in cercarial tails than whole cercariae or schistosomula while the opposite pattern is found for glucose transporters and hexokinase transcripts; (2) Schistosomula contain low levels of transcripts encoding respiratory enzymes but regain the capacity for aerobic glucose metabolism as they mature to adulthood; (3) Male and female adults contain similar levels of the different transcripts involved in glucose metabolism.

Novel mechanisms of immune evasion by Schistosoma mansoni

The interaction of Schistosoma mansoni with its host's immune system is largely affected by multiple specific and non-specific evasion mechanisms employed by the parasite to reduce the host's immune reactivity. Only little is known about these mechanisms on the molecular level. The four molecules described below are intrinsic parasitic proteins recently identified and studied in our laboratory. 1. m28-A 28kDa membrane serine protease. m28 cleaves iC3b and can thus restrict attack by effector cells utilizing complement receptors (especially CR3). Treatment with protease inhibitors potentiates killing of schistosomula by complement plus neutrophils. 2. Smpi56-A 56kDa serine protease inhibitor. Smpi56 binds covalently to m28 and to neutrophil's elastase and blocks their proteolytic activity. 3. P70-A 70kDa C3b binding protein. The postulated activity of P70 includes binding to C3b and blocking of complement activation of the C3 step. 4. SCIP-1-A 94kDa schistosome complement inhibitor. SCIP-1 shows antigenic and functional similarities to the human 18kDa complement inhibitor CD59. Like CD59, SCIP-1 binds to C8 and C9 and blocks formation of the complement membrane attack complex. Antibodies directed to human CD59 bind to schistosomula and potentiate their killing by complement. The structure and function of these four proteins as well as their capacity to induce protection from infection with S. mansoni are under investigation.

Neotropical Monogenoidea. 24. Rhinoxenus bulbovaginatus n. sp. (Dactylogyridae, Ancyrocephalinae) from the nasal cavity of Salminus maxillosus (Osteichthyes, Characidae) from the Rio Paraná, Paraná, Brazil

Rhinoxenus bulbovaginatus n. sp. is described from the nose of Salminus maxillosus (Characidae) collected in the basin of the rio Paraná, near the city of Porto Rico, state of Paraná, Brazil. The new species can be differentiated from the other three species in the genus by the morphology of the copulatory complex, vagina, and ventral anchor. The sister group relationship of the known species of Rhinoxenus was determined using techniques of Phylogenetic Systematics (Cladism). The resulting cladogram (C.I.=100%) indicates that the new species is most closely related to R. piranhus Kritsky, Boeger and Thatcher, 1988. The other two species of the genus, R. arietinus Kritsky, Boeger and Thatcher, 1988 and R. nyttus Kritsky, Boeger and Thatcher, 1988, both parasites of Anostomidae fishes, have a paraphyletic position in the cladogram, suggesting that the origin of at least one of them can not be associated to cospeciation.