Basic biochemical investigations as rationale for the design of original antimalarial drugs. An example of phospholipid metabolism

The future of antimalarial chemotherapy is particulary alarming in view of the spread of parasite cross-resistances to drugs that are not even structurally related. Only the availability of new pharmacological models will make it possible to select molecules with novel mechanisms of action, thus delaving resistance and allowing the development of new chemotherapeutic strategies. We reached this objective in mice. Our approach is hunged on fundamental and applied research begun in 1980 to investigate to phospholipid (PL) metabolism of intraerythrocytic Plasmodium. This metabolism is abundant, specific and indispensable for the production of Plasmodium membranes. Any drug to interfere with this metabolism blocks parasitic development. The most effective interference yet found involves blockage of the choline transporter, which supplies Plasmodium with choline for the synthesis of phosphatidylcholine, its major PL, this is a limiting step in the pathway. The drug sensitivity thereshold is much lower for the parasite, which is more dependent on this metabolism than host cells. The compounds show in vitro activity against P. falciparum at 1 to 10 nM. They show a very low toxicity against a lymphblastoid cell line, demonstrating a total abscence of correlation between growth inhibition of parasites and lymphoblastoid cells. They show antimalarial activity in vivo, in the P. berghei or P. chabaudi/mouse system, at doses 20-to 100-fold lower than their in acute toxicity limit. The bioavailability of a radiolabeled form of the product seemed to be advantageous (slow blood clearance and no significant concentration in tissues). Lastly, the compounds are inexpensive to produce. They are stable and water-soluble.

Biogeography of Triatominae (Hemiptera: Reduviidae) in Ecuador: implications for the design of control strategies

Chagas disease control strategies strongly depend on the triatomine vector species involved in Trypanosoma cruzi transmission within each area. Here we report the results of the identification of specimens belonging to various species of Triatominae captured in Ecuador (15 species from 17 provinces) and deposited in the entomological collections of the Catholic University of Ecuador (Quito), Instituto Oswaldo Cruz (Brazil), the Natural History Museum London (UK), the London School of Hygiene and Tropical Medicine (UK), the National Institute of Hygiene (Quito), and the Vozandes Hospital (Quito). A critical review of published information and new field records are presented. We analysed these data in relation to the life zones where triatomines occur (11 life zones, excluding those over 2,200 m altitude), and provide biogeographical maps for each species. These records are discussed in terms of epidemiological significance and design of control strategies. Findings relevant to the control of the main vector species are emphasised. Different lines of evidence suggest that Triatoma dimidiata is not native to Ecuador-Peru, and that synanthropic populations of Rhodnius ecuadoriensis in southern Ecuador-northern Peru might be isolated from their sylvatic conspecifics. Local eradication of T. dimidiata and these R. ecuadoriensis populations might therefore be attainable. However, the presence of a wide variety of native species indicates the necessity for a strong longitudinal surveillance system.

Polymorphism of the Human Immunodeficiency Virus Type 1 in Brazil: Genetic Characterization of the nef Gene and Implications for Vaccine Design

Most of the Brazilian HIV-1 samples have been characterized based on the structural genes (env, gag and pol) and no data concerning the variability of the accessory genes such as nef have been available so far. Considering the role of the nef on virus biology and the inclusion of this region in some HIV/AIDS vaccine products under testing, the purpose of this study was to document the genetic diversity of the nef gene in third-four HIV-1 Brazilian samples previously subtyped based on the env C2-V3 region. Although only few non-subtype B samples have already been analyzed so far, the cytotoxic Tlymphocyte epitopes encoded in this region were relatively conserved among the subtypes, with some amino acid signatures mainly in the subtype C samples. Considering the increasing of the non-B HIV-1 subtypes worldwide, in special the subtype C, more data should be generated concerning the genetic and antigenic variability of these subtypes, as well as the study of the impact of such polymorphism in HIV/AIDS vaccine design and testing.

Mapping the molecular characteristics of Brazilian human T-cell lymphotropic virus type 1 Env (gp46) and Pol amino acid sequences for vaccine design

This study was carried out to evaluate the molecular pattern of all available Brazilian human T-cell lymphotropic virus type 1 Env (n = 15) and Pol (n = 43) nucleotide sequences via epitope prediction, physico-chemical analysis, and protein potential sites identification, giving support to the Brazilian AIDS vaccine program. In 12 previously described peptides of the Env sequences we found 12 epitopes, while in 4 peptides of the Pol sequences we found 4 epitopes. The total variation on the amino acid composition was 9 and 17% for human leukocyte antigen (HLA) class I and class II Env epitopes, respectively. After analyzing the Pol sequences, results revealed a total amino acid variation of 0.75% for HLA-I and HLA-II epitopes. In 5 of the 12 Env epitopes the physico-chemical analysis demonstrated that the mutations magnified the antigenicity profile. The potential protein domain analysis of Env sequences showed the loss of a CK-2 phosphorylation site caused by D197N mutation in one epitope, and a N-glycosylation site caused by S246Y and V247I mutations in another epitope. Besides, the analysis of selection pressure have found 8 positive selected sites (w = 9.59) using the codon-based substitution models and maximum-likelihood methods. These studies underscore the importance of this Env region for the virus fitness, for the host immune response and, therefore, for the development of vaccine candidates.

A clinical trial for uniform multidrug therapy for leprosy patients in Brazil: rationale and design

Leprosy will continue to be a public health problem for several decades. The World Health Organization (WHO) recommends that, for treatment purposes, leprosy cases be classified as either paucibacillary or multibacillary (MB). A uniform leprosy treatment regimen would simplify treatment and halve the treatment duration for MB patients. The clinical trial for uniform multidrug therapy (U-MDT) for leprosy patients (LPs) in Brazil is a randomised, open-label clinical trial to evaluate if the effectiveness of U-MDT for leprosy equals the regular regimen, to determine the acceptability of the U-MDT regimen and to identify the prognostic factors. This paper details the clinical trial methodology and patient enrolment data. The study enrolled 858 patients at two centres and 78.4% of participants were classified as MB according to the WHO criteria. The main difficulty in evaluating a new leprosy treatment regimen is that no reliable data are available for the current treatment regimen. Relapse, reaction and impaired nerve function rates have never been systematically determined, although reaction and impaired nerve function are the two major causes of nerve damage that lead to impairments and disabilities in LPs. Our study was designed to overcome the need for reliable data about the current treatment and to compare its efficacy with that of a uniform regimen.

Structure-based drug design studies of the interactions ofent-kaurane diterpenes derived from Wedelia paludosa with the Plasmodium falciparumsarco/endoplasmic reticulum Ca2+-ATPase PfATP6

Malaria is responsible for more deaths around the world than any other parasitic disease. Due to the emergence of strains that are resistant to the current chemotherapeutic antimalarial arsenal, the search for new antimalarial drugs remains urgent though hampered by a lack of knowledge regarding the molecular mechanisms of artemisinin resistance. Semisynthetic compounds derived from diterpenes from the medicinal plant Wedelia paludosawere tested in silico against the Plasmodium falciparumCa2+-ATPase, PfATP6. This protein was constructed by comparative modelling using the three-dimensional structure of a homologous protein, 1IWO, as a scaffold. Compound 21 showed the best docking scores, indicating a better interaction with PfATP6 than that of thapsigargin, the natural inhibitor. Inhibition of PfATP6 by diterpene compounds could promote a change in calcium homeostasis, leading to parasite death. These data suggest PfATP6 as a potential target for the antimalarial ent-kaurane diterpenes.

Computational drug design strategies applied to the modelling of human immunodeficiency virus-1 reverse transcriptase inhibitors

Reverse transcriptase (RT) is a multifunctional enzyme in the human immunodeficiency virus (HIV)-1 life cycle and represents a primary target for drug discovery efforts against HIV-1 infection. Two classes of RT inhibitors, the nucleoside RT inhibitors (NRTIs) and the nonnucleoside transcriptase inhibitors are prominently used in the highly active antiretroviral therapy in combination with other anti-HIV drugs. However, the rapid emergence of drug-resistant viral strains has limited the successful rate of the anti-HIV agents. Computational methods are a significant part of the drug design process and indispensable to study drug resistance. In this review, recent advances in computer-aided drug design for the rational design of new compounds against HIV-1 RT using methods such as molecular docking, molecular dynamics, free energy calculations, quantitative structure-activity relationships, pharmacophore modelling and absorption, distribution, metabolism, excretion and toxicity prediction are discussed. Successful applications of these methodologies are also highlighted.

Mortalidade infantil em município do interior do estado de São Paulo

Considerando que a mortalidade infantil é indicador dos níveis de saúde da população, realizamos este trabalho, cujo objetivo foi identificar as causas de mortalidade infantil no ano de 1998 em Botucatu. O coeficiente de mortalidade infantil obtido foi 12/1000 NV, com maior participação dos óbitos neonatais - 8,3/1000 NV A maior parte dos óbitos foi classificada como reduzível ou parcialmente reduzível, mas a atenção necessária para viabilizar tal redução foi variada. Dos óbitos ocorridos, 21,7% eram inevitáveis, evidenciando que para redução dos índices de mortalidade infantil deveremos continuar investindo na qualidade da assistência à saúde e melhoria das condições de vida da população.

Representação de gestantes tabagistas sobre o uso do cigarro: estudo realizado em hospital do interior paulista

Este estudo objetivou apreender as representações de gestantes tabagistas sobre o uso de cigarro. Utilizou-se como referencial teórico a Teoria das Representações Sociais. Para análise dos dados, construiu-se o Discurso do Sujeito Coletivo. Das 27 mulheres entrevistadas, 18 possuíam primeiro grau completo, oito, o segundo grau completo e uma, ensino superior; 14 tinham união estável, seis eram casadas. Quatro temas emergiram: 1) o início do hábito de fumar: prática social e natural; 2) satisfação versus culpa; 3) uma bomba: efeitos do cigarro na gestação; 4) cessação: entre o querer e o poder. Apreendeu-se representação negativa do cigarro, considerado o pior dos vícios e potencial causador de complicações feto-maternas. O tabagismo foi representado de maneira preconceituosa, desconsiderando a existência e necessidade de tratamento. Emergiram dificuldades relativas à cessação, trazendo a necessidade de ajuda profissional, para informações, abordagem e tratamento adequados e apoio para que se alcance êxito.

Qualidade de vida de mulheres vivendo com o HIV/aids de um município do interior paulista

A terapia antirretroviral de alta potência beneficia os indivíduos com HIV/aids na sobrevida, cronicidade e qualidade de vida. Este estudo de corte transversal, com abordagem quantitativa, objetivou avaliar a qualidade de vida de mulheres com HIV/aids, utilizando o WHOQOL - HIV bref e sua associação com variáveis sociodemográficas. Foi realizado em dois ambulatórios especializados no atendimento a indivíduos com HIV/aids. De 106 mulheres participantes, 99,1% eram heterossexuais e 92,4% foram infectadas por via sexual. Dentre os domínios de qualidade de vida, espiritualidade obteve maior escore (65,7), seguido pelo físico (64,7), psicológico (60,6), relações sociais (59,5). Menores escores foram atingidos nos domínios nível de independência (58,6) e meio ambiente (54,5). Evidenciou-se que os fatores baixo nível socioeconômico e educacional tiveram associação com diferentes domínios, denotando a relação entre qualidade de vida e condições de vida. Concluiu-se que persistem os desafios no âmbito das relações sociais, afetivas, financeiras, requerendo intervenções efetivas focando o empoderamento das mulheres com HIV/aids.

O monitoramento de processos físicos de esterilização em hospitais do interior do estado de Goiás

Estudo descritivo com o objetivo de identificar a realização de controles físicos, químicos e biológicos dos processos de esterilização pelo vapor saturado sob pressão e em estufas de Pasteur em Centros de Material e Esterilização - CME. Os dados foram obtidos por meio de entrevista ao responsável pelo CME do maior hospital de todas as cidades do interior do Estado de Goiás, com número de habitantes igual ou superior a 20.000, no período de agosto de 2005 a junho de 2006. Participaram 44 municípios. Foi utilizado o programa SPSS, para análise. Em 31 (94,0%) hospitais não havia enfermeiros exclusivos no CME, os responsáveis eram técnicos e auxiliares de enfermagem. A maioria não realizava os controles físicos, químicos e biológicos dos processos de esterilização e, em apenas um, esses eram realizados simultaneamente. O descumprimento da monitorização dos ciclos de esterilização, impedindo a garantia da qualidade dos processos, representa risco à população assistida.

Abundância e riqueza de espécies de Syrphidae (Diptera) em áreas de borda e interior de floresta no Parque Estadual de Vila Velha, Ponta Grossa, Paraná, Brasil

Com o objetivo de caracterizar a fauna local de insetos foram obtidas amostras semanais, de setembro/1999 a agosto/2000, utilizando-se armadilhas Malaise instaladas na borda da floresta e no seu interior. Uma análise temporal foi realizada com as espécies de Syrphidae coletadas há, aproximadamente, dezessete anos no mesmo local, dentro da floresta. A abundância e a riqueza de espécies também foram avaliadas. Tanto a riqueza quanto a abundância foram maiores na borda da floresta. Comparando-se os dados atuais com aqueles obtidos em 1986/1987, observa-se um decréscimo na abundância e também na riqueza de espécies de Syrphidae. A espécie mais abundante na borda foi Allograpta neotropica Curran, 1936 e no interior (1999/2000), Ocyptamus sativus (Curran, 1941). Os espécimens de Toxomerus Macquart, 1855 foram os mais abundantes na armadilha localizada na borda da floresta e os de Ocyptamus Macquart, 1834 no interior. Noventa e cinco espécies foram identificadas em 22 gêneros. Ocyptamus foi o gênero com maior riqueza de espécies (23). Na seqüência estão Copestylum Macquart, 1846 (15), Toxomerus (15) e Microdon Meigen, 1803 (10). Sete espécies foram comuns aos três levantamentos: Allograpta neotropica; Copestylum selectum (Curran, 1939); Leucopodella gracilis (Williston, 1891); Mixogaster polistes Hull, 1954; Ocyptamus funebris Macquart, 1834; Toxomerus procrastinatus Metz, 2001 e Toxomerus tibicen (Wiedemann, 1830). Três novas espécies de Microdon, uma de Toxomerus, uma de Aristosyrphus Curran, 1941 e uma de Myolepta Newman, 1838 foram identificadas.

Uma nova subespécie de Parides bunichus (Hübner) (Lepidoptera, Papilionidae, Troidini) do interior da Bahia, Brasil

Parides bunichus almas ssp.nov. é descrita com base em material coletado no Pico das Almas, Rio de Contas, Bahia, Brasil.