The sequence of a 200 kb portion of a Plasmodium vivax chromosome reveals a high degree of conservation with Plasmodium falciparum chromosome 3

Within a 199 866 base pair (bp) portion of a Plasmodium vivax chromosome we identified a conserved linkage group consisting of at least 41 genes homologous to Plasmodium falciparum genes located on chromosome 3. There were no P. vivax homologues of the P. falciparum cytoadherence-linked asexual genes clag 3.2, clag 3.1 and a var C pseudogene found on the P. vivax chromosome. Within the conserved linkage group, the gene order and structure are identical to those of P. falciparum chromosome 3. This conserved linkage group may extend to as many as 190 genes. The subtelomeric regions are different in size and the P. vivax segment contains genes for which no P. falciparum homologues have been identified to date. The size difference of at least 900 kb between the homologous P. vivax chromosome and P. falciparum chromosome 3 is presumably due to a translocation. There is substantial sequence divergence with a much higher guanine + cytosine (G + C) content in the DNA and a preference for amino acids using GC-rich codons in the deduced proteins of P. vivax. This structural conservation of homologous genes and their products combined with sequence divergence at the nucleotide level makes the P. vivax genome a powerful tool for comparative analyses of Plasmodium genomes. (C) 2001 Elsevier Science B.V. All rights reserved.

Complete DNA sequence and gene organization of the mitochondrial genome of the liverfluke, Fasciola hepatica L. (Platyhelminthes; Trematoda)

The complete nucleotide sequence of the mitochondrial (mt) DNA molecule of the liverfluke, Fasciola hepatica (phylum Platyhelminthes, class Trematoda, family Fasciolidae), was determined, It comprises 14462 bp, contains 12 protein-encoding, 2 ribosomal and 22 transfer RNA genes, and is the second complete flatworm (and the first trematode) mitochondrial sequence to be described in detail. All of the genes are transcribed from the same strand. Of the genes typically found in mitochondrial genomes of eumetazoans, only atp8 is absent. The nad4L and nad4 genes overlap by 40 nt. Most intergenic sequences are very short. Two larger non-coding regions are present. The longer one (817 nt) is located between trnG and cox3 and consists of 8 identical tandem repeats of 85 nt, rich in G and C, followed by 1 imperfect repeat. The shorter non-coding region (187 nt) exhibits no special features and is separated from the longer region by trnG. The gene arrangement resembles that of some other trematodes including the eastern Asian Schistosoma species (and cyclophyllidean cestode species) but it is strikingly different from that of the African schistosomes, represented by Schistosoma mansoni. The genetic code is as inferred previously for flatworms. Transfer RNA genes range in length from 58 to 70 nt, their products producing characteristic 'clover leaf' structures, except for tRNA(S-VON) and tRNA(S-AGN) lacking the DHU arm.

Immune responses in hookworm infections

Hookworms infect perhaps one-fifth of the entire human population, yet little is known about their interaction with our immune system. The two major species are Necator americanus, which is adapted to tropical conditions, and Ancylostoma duodenale, which predominates in more temperate zones. While having many common features, they also differ in several key aspects of their biology. Host immune responses are triggered by larval invasion of the skin, larval migration through the circulation and lungs, and worm establishment in the intestine, where adult worms feed on blood and mucosa while injecting various molecules that facilitate feeding and modulate host protective responses. Despite repeated exposure, protective immunity does not seem to develop in humans, so that infections occur in all age groups (depending on exposure patterns) and tend to be prolonged. Responses to both larval and adult worms have a characteristic T-helper type 2 profile, with activated mast cells in the gut mucosa, elevated levels of circulating immunoglobulin E, and eosinoophilia in the peripheral blood and local tissues, features also characteristic of type I hypersensitivity reactions. The longevity of adult hookworms is determined probably more by parasite genetics than by host immunity. However, many of the proteins released by the parasites seem to have immunomodulatory activity, presumably for self-protection. Advances in molecular biotechnology enable the identification and characterization of increasing numbers of these parasite molecules and should enhance our detailed understanding of the protective and pathogenetic mechanisms in hookworm infections.

Apoptotic deletion of Th cells specific for the 19-kDa carboxyl-terminal fragment of merozoite surface protein 1 during malaria infection

Immunity induced by the 19-kDa fragment of merozoite surface protein 1 is dependent on CD4(+) Th cells. However, we found that adoptively transferred CFSE-labeled Th cells specific for an epitope on Plasmodium yoelii 19-kDa fragment of merozoite surface protein 1 (peptide (p)24), but not OVA-specific T cells, were deleted as a result of P. yoelii infection. As a result of infection, spleen cells recovered from infected p24-specific T cell-transfused mice demonstrated reduced response to specific Ag. A higher percentage of CFSE-labeled p24-specific T cells stained positive with annexin and anti-active caspase-3 in infected compared with uninfected mice, suggesting that apoptosis contributed to deletion of p24-specific T cells during infection. Apoptosis correlated with increased percentages of p24-specific T cells that stained positive for Fas from infected mice, suggesting that P. yoelii-induced apoptosis is, at least in part, mediated by Fas. However, bystander cells of other specificities also showed increased Fas expression during infection, suggesting that Fas expression alone is not sufficient for apoptosis. These data have implications for the development of immunity in the face of endemic parasite exposure.

Nuclear PTEN expression and clinicopathologic features in a population-based series of primary cutaneous melanoma

Germline mutations of the PTEN tumor-suppressor gene, on 10q23, cause Cowden syndrome, an inherited hamartoma syndrome with a high risk of breast, thyroid and endometrial carcinomas and, some suggest, melanoma. To date, most studies which strongly implicate PTEN in the etiology of sporadic melanomas have depended on cell lines, short-term tumor cultures and noncultured metastatic melanomas. The only study which reports PTEN protein expression in melanoma focuses on cytoplasmic expression, mainly in metastatic samples. To determine how PTEN contributes to the etiology or the progression of primary cutaneous melanoma, we examined cytoplasmic and nuclear PTEN expression against clinical and pathologic features in a population-based sample of 150 individuals with incident primary cutaneous melanoma. Among 92 evaluable samples, 30 had no or decreased cytoplasmic PTEN protein expression and the remaining 62 had normal PTEN expression. In contrast, 84 tumors had no or decreased nuclear expression and 8 had normal nuclear PTEN expression. None of the clinical features studied, such as Clark's level and Breslow thickness or sun exposure, were associated with cytoplasmic PTEN expressional levels. An association with loss of nuclear PTEN expression was indicated for anatomical site (p = 0.06) and mitotic index (p = 0.02). There was also an association for melanomas to either not express nuclear PTEN or to express p53 alone, rather than both simultaneously (p = 0.02). In contrast with metastatic melanoma, where we have shown previously that almost two-thirds of tumors have some PTEN inactivation, only one-third of primary melanomas had PTEN silencing. This suggests that PTEN inactivation is a late event likely related to melanoma progression rather than initiation. Taken together with our previous observations in thyroid and islet cell tumors, our data suggest that nuclear-cytoplasmic partitioning of PTEN might also play a role in melanoma progression. (C) 2002 Wiley-Liss, Inc.

Veterinary parasitology teaching in eastern Australia

There are tendencies in universities globally to change undergraduate teaching in veterinary parasitology. To be able to give considered advice to universities, faculties, governmental bodies and professional societies about a discipline and to establish how particular changes may impact on the quality of a course, is the requirement to record and review its current status. The present paper contributes toward this objective by providing a snap-shot of the veterinary parasitology courses at the Universities of Melbourne, Sydney and Queensland in eastern Australia. It includes a description of the veterinary science curriculum in each institution, and provides an outline of its veterinary parasitology course, including objectives, topics covered, course delivery, student examination procedures and course evaluation. Student contact time in veterinary parasitology during the curriculum is currently higher in Melbourne (183 h) compared with Sydney and Queensland (106-110 h). In the teaching of parasitology, Melbourne adopts a taxonomic approach (in the pre-clinical period) followed by a combined disciplinary and problem-based approach in the clinical semesters, whereas both Sydney and Queensland focus more on presenting parasites on a host species-basis followed by a problem-based approach. (C) 2002 Elsevier Science B.V. All rights reserved.

The triad of bilateral retinoblastoma, dysplastic naevus syndrome and multiple cutaneous malignant melanomas: a case report and review of the literature

We report a case of a patient with the triad of retinoblastoma, dysplastic naevus syndrome (DNS) and multiple cutaneous melanomas. The combination of retinoblastoma and DNS is a significant risk factor for the development of cutaneous melanoma. This risk extends to family members. We recommend that survivors of (inherited) retinoblastoma and their relatives are closely screened for the presence of dysplastic naevi. (C) 2002 Lippincott Williams Wilkins.

Secretory pathway of trypanosomatid parasites

The Trypanosomatidae comprise a large group of parasitic protozoa, some of which cause important diseases in humans. These include Tryanosoma brucei (the causative agent of African sleeping sickness and nagana in cattle), Trypanosoma cruzi (the causative agent of Chagas' disease in Central and South America), and Leishmania spp. (the causative agent of visceral and [muco]cutaneous leishmaniasis throughout the tropics and subtropics). The cell surfaces of these parasites are covered in complex protein- or carbohydrate-rich coats that are required for parasite survival and infectivity in their respective insect vectors and mammalian hosts. These molecules are assembled in the secretory pathway. Recent advances in the genetic manipulation of these parasites as well as progress with the parasite genome projects has greatly advanced our understanding of processes that underlie secretory transport in trypanosomatids. This article provides an overview of the organization of the trypanosomatid secretory pathway and connections that exist with endocytic organelles and multiple lytic and storage vacuoles. A number of the molecular components that are required for vesicular transport have been identified, as have some of the sorting signals that direct proteins to the cell surface or organelles it? the endosome-vacuole system. Finally, the subcellular organization of the major glycosylation pathways in these parasites is reviewed. Studies on these highly divergent eukaryotes provide important insights into the molecular processes underlying secretory transport that arose very early in eukaryotic evolution. They also reveal unusual or novel aspects of secretory), transport and protein glycosylation that may be exploited in developing new antiparasite drugs.

Melanocytic nevi, solar keratoses, and divergent pathways to cutaneous melanoma

Background: Some melanomas form on sun-exposed body sites, whereas others do not. We previously proposed that melanomas at different body sites arise through different pathways that have different associations with melanocytic nevi and solar keratoses. We tested this hypothesis in a case-case comparative study of melanoma patients in Queensland, Australia. Methods: We randomly selected patients from among three prespecified groups reported to the population-based Queensland Cancer Registry: those with superficial spreading or nodular melanomas of the trunk (n = 154, the reference group), those with such melanomas of the head and neck (n = 77, the main comparison group), and those with lentigo maligna melanoma (LMM) (n = 75, the chronic sun-exposed group). Each participant completed a questionnaire, and a research nurse counted melanocytic nevi and solar keratoses. We calculated exposure odds ratios (ORs) and 95% confidence intervals (CIs) to quantify the association between factors of interest and each melanoma group. Results: Patients with head and neck melanomas, compared with patients with melanomas of the trunk, were statistically significantly less likely to have more than 60 nevi (OR = 0.34, 95% CI = 0.15 to 0.79) but were statistically significantly more likely to have more than 20 solar keratoses (OR = 3.61, 95% CI = 1.42 to 9.17) and also tended to have a past history of excised solar skin lesions (OR = 1.87, 95% CI = 0.89 to 3.92). Patients with LMM were also less likely than patients with truncal melanomas to have more than 60 nevi (OR = 0.32, 95% CI = 0.14 to 0.75) and tended toward more solar keratoses (OR = 2.14, 95% CI = 0.88 to 5.16). Conclusions: Prevalences of nevi and solar keratoses differ markedly between patients with head and neck melanomas or LMM and patients with melanomas of the trunk. Cutaneous melanomas may arise through two pathways, one associated with melanocyte proliferation and the other with chronic exposure to sunlight.

Análise de fatores geográficos e climáticos relacionados à distribuição de Phlebotominae (Diptera: Psychodidae) em área de transmissão de leishmaniose tegumentar americana no estado do Espírito Santo, Brasil

A leishmaniose tegumentar americana (LTA) é uma doença que acomete pele e mucosas causada por parasitos dermotrópicos do gênero Leishmania Ross, 1903. Os parasitos são transmitidos através da picada de pequenos dípteros da família Psychodidae, os flebotomíneos. O município de Cariacica, Espírito Santo, Brasil, esteve nos últimos cinco anos (2009 a 2013) entre os cinco que apresentaram maior número de casos notificados no estado, segundo a Secretaria de Estado de Saúde (SESA-ES, 2014). A localidade de Roda D’Água demonstra grande importância, por concentrar elevado número de casos, contribuindo com grande parte das notificações do município. Avaliando os casos da doença na região, registrados nos prontuários médicos do serviço de referência, na Unidade de Medicina Tropical da Universidade Federal do Espírito Santo, observou-se que estes ocorriam a até 550 m de altitude, numa área que vai de 20 a 718 m. A hipótese mais provável seria a de que o fenômeno fosse relacionado aos vetores, já que o homem e os animais estariam presentes em todas as altitudes. De fevereiro de 2002 a janeiro de 2004 foram realizadas coletas mensais de flebotomíneos em Roda D’Água, que aconteciam simultaneamente em três níveis de altitude, sendo: nível 1 - até 250 m; nível 2 - entre 250 e 500m e nível 3 - acima de 500m. Em cada nível as coletas aconteciam em dois ambientes: mata e peridomicílio. As capturas eram feitas em armadilhas de Shannon modificadas e por busca ativa em repouso, com capturador de Castro. Avaliou-se o comportamento das espécies quanto à pluviosidade (períodos seco e chuvoso) e às estações do ano. Analisaram-se estatisticamente as principais espécies antropofílicas de importância epidemiológica (Falqueto, 1995). Foram calculados os índices ecológicos abundância, riqueza, diversidade, equitabilidade e dominância. Coletou-se um total de 13233 flebotomíneos, com identificação de 23 espécies. A espécie predominante foi Nyssomyia intermedia (61,12%), seguida por Pintomyia fischeri (18,20%) e Migonemyia migonei (8,68%), todas antropofílicas. Somou-se a estas a espécie Pintomyia monticola, que representou 1,67% do total de espécimes coletados e também é altamente antropofílica. As demais espécies somaram 10,10% do total de flebotomineos. A altitude influenciou a distribuição das quatro espécies analisadas, tendo Ny. intermedia e Pi. fischeri sido mais abundantes no nível 2, Mg. migonei mais abundante no nível 1 e Pi. monticola no nível 3. Em relação ao ambiente, as espécies Ny. intermedia e Mg. migonei foram estatisticamente mais abundantes no peridomicílio e Pi. monticola na mata. A distribuição de Pi. fisheri não apresentou diferença significativa entre os dois ambientes, porém foi a única afetada pelas chuvas e estações do ano, sendo a espécie mais encontrada no período seco e no inverno. Nyssomyia intermedia parece ser a principal espécie vetora da LTA em Roda D’Água, com Mg. migonei provavelmente tendo papel secundário. Pi. fisheri não parece estar envolvido localmente na transmissão da doença para humanos, apesar de já ter sido incriminado em outras regiões. A distribuição de Pi. monticola em relação à altitude e ao ambiente indica ser improvável sua participação na transmissão da LTA naquela região.

Description of Lutzomyia (Pifanomyia) robusta n. sp. (Diptera, Psychodidae, Phlebotominae) from Peruvian Equadorean interandean areas

Description of Lutzomyia robusta, n. sp. (Diptera, Psychodidae, Phlebotominae) from interandean areas of Peru and Equador. Lutzomyia robusta, n. sp., probable vector of human bartonellosis and cutaneous leishmaniasis, is described and illustrated. This species presents strong affinity with L. serrana (Damasceno & Arouck, 1949) but they can be distinguished by variance analysis of four male characteristics and only one female characteristic. In the variance analysis, populations of L. serrana, of Amazonian areas of Brazil, Peru and Bolivia, the coast of Equador and other areas of Brazil were studied. The synonymy of Lutzomyia guayasi (Rodriguez) and L. serrana was corroborated.

Detection of anti-Giardia lamblia serum antibody among children of day care centers

OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA) for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3%) showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3%) and 100 (68%) samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39%) than IIF (70%). IIF also showed a higher concordance with microscopic examination than ELISA.

Exposure to larva migrans syndromes in squares and public parks of cities in Chile

Between November 2001 and December 2002, 600 dog fecal samples were collected in main squares and public parks of 13 cities in Chile, from the extreme north to the extreme south of the country. The samples were processed in the laboratory by centrifugal sedimentation and the Harada-Mori methods. T. canis eggs were found in 12 cities. Detection rates ranged from 1.9 to 12.5% with an average of 5.2%. Seven percent of the samples had eggs and 9.5% had rhabditoid and/or filariform larvae of Ancylostomatidae. Strongyloides stercoralis were not found. Squares and public parks in Chile pose a potential risk of exposure to visceral, ocular, and/or cutaneous larva migrans syndromes.

Phlebotomine fauna in a rural area of the Brazilian Pantanal

The aim of the study was to identify among the phlebotomine fauna potential leishmaniasis vectors. The study was carried out in Corumbá county, State of Mato Grosso do Sul, Mid-West Brazil (18º59'S, 56º39'W). Sand fly captures were undertaken fortnightly with automatic light traps at 11 sites in forested environments and anthropic areas from April 2001 to July 2003. A total of only 41 specimens were captured. Thirty-one percent of the specimens were captured in forests and 68.3% in anthropic areas. The predominance of non-anthropophilic groups and the low density of N. whitmani, a known cutaneous leishmaniasis vector, does not seem to indicate any actual risk of the transmission of this disease in the study area.