Trypanosoma cruzi ribosomal protein S4: characterization of its coding locus, analysis of transcripts, and antigenicity of the protein

Two allelic genomic fragments containing ribosomal protein S4 encoding genes (rpS4) from Trypanosoma cruzi (CL-Brener strain) were isolated and characterized. One allele comprises two complete tandem repeats of a sequence encoding an rpS4 gene. In the other, only one rpS4 gene is found. Sequence comparison to the accessed data in the genome project database reveals that our two-copy allele corresponds to a variant haplotype. However, the deduced aminoacid sequence of all the gene copies is identical. The rpS4 transcripts processing sites were determined by comparison of genomic sequences with published cDNA data. The obtained sequence data demonstrates that rpS4 genes are expressed in epimastigotes, amastigotes, and trypomastigotes. A recombinant version of rpS4 was found to be an antigenic: it was recognized by 62.5% of the individuals with positive serology for T. cruzi and by 93.3% of patients with proven chronic chagasic disease.

Anti-leishmanial and structure-activity relationship of ring substituted 3-phenyl-1-(1,4-di-N-oxide quinoxalin-2-yl)-2-propen-1-one derivatives

A series of ring substituted 3-phenyl-1-(1,4-di-N-oxide quinoxalin-2-yl)-2-propen-1-one derivatives were synthesized and tested for in vitro leishmanicidal activity against amastigotes of Leishmania amazonensis in axenical cultures and murine infected macrophages. Structure-activity relationships demonstrated the importance of a radical methoxy at position R3', R4' and R5'. (2E)-3-(3,4,5-trimethoxy-phenyl)-1-(3,6,7-trimethyl-1,4-dioxy-quinoxalin-2-yl)-propenone was the most active. Cytotoxicity on macrophages revealed that this product was almost six times more active than toxic.

Performance levels of four Latin American laboratories for the serodiagnosis of Chagas disease in Mexican sera samples

In nearly all of the previous multicentre studies evaluating serological tests for Trypanosoma cruzi infection, sera samples from Central or South American countries have been used preferentially. In this work we compared the reliability of the serological tests using Mexican sera samples that were evaluated in four independent laboratories. This included a reference laboratory in Brazil and three participant laboratories, including one in Central America and two in Mexico. The kappa index between Brazilian and Honduran laboratories reached 1.0 and the index for the Mexican laboratories reached 0.94. Another finding of this study was that the source of antigen did not affect the performance of the serological tests.

Distribution and infection of triatomines (Hemiptera: Reduviidae) by Trypanosoma cruzi in the state of Michoacán, Mexico

An entomological study of triatomine species was carried out to assess their prevalence in 10 localities of the state of Michoacán, Mexico. Entomological indices were calculated to estimate the risk for vector-borne transmission of Trypanosoma cruzi to the human population in this area. Four triatomine species (Triatoma barberi, Triatoma dimidiata, Meccus pallidipennis and Meccus longipennis) were collected from the study area. This is the first report of M. longipennis and T. dimidiata in Michoacán. M. pallidipennis was significantly (p < 0.05) more abundant than any of the other species collected in the study area. Infection indices were greater than 50% for each of the four collected triatomine species. Significantly more triatomines were collected from intradomiciliary areas than from peridomiciliary or sylvatic areas. Infestation, crowding and density indices were low, whereas colonisation indices were high in five localities. The current vectorial conditions in the study area require continuous entomological and serological surveillance to diminish the risk of T. cruzi transmission to human populations.

Dengue-2 and yellow fever 17DD viruses infect human dendritic cells, resulting in an induction of activation markers, cytokines and chemokines and secretion of different TNF-α and IFN-α profiles

Flaviviruses cause severe acute febrile and haemorrhagic infections, including dengue and yellow fever and the pathogenesis of these infections is caused by an exacerbated immune response. Dendritic cells (DCs) are targets for dengue virus (DENV) and yellow fever virus (YF) replication and are the first cell population to interact with these viruses during a natural infection, which leads to an induction of protective immunity in humans. We studied the infectivity of DENV2 (strain 16681), a YF vaccine (YF17DD) and a chimeric YF17D/DENV2 vaccine in monocyte-derived DCs in vitro with regard to cell maturation, activation and cytokine production. Higher viral antigen positive cell frequencies were observed for DENV2 when compared with both vaccine viruses. Flavivirus-infected cultures exhibited dendritic cell activation and maturation molecules. CD38 expression on DCs was enhanced for both DENV2 and YF17DD, whereas OX40L expression was decreased as compared to mock-stimulated cells, suggesting that a T helper 1 profile is favoured. Tumor necrosis factor (TNF)-α production in cell cultures was significantly higher in DENV2-infected cultures than in cultures infected with YF17DD or YF17D/DENV. In contrast, the vaccines induced higher IFN-α levels than DENV2. The differential cytokine production indicates that DENV2 results in TNF induction, which discriminates it from vaccine viruses that preferentially stimulate interferon expression. These differential response profiles may influence the pathogenic infection outcome.

How to increase the population of a Phlebotomus perniciosus (Diptera: Psychodidae) colony: a new method

The sandfly Phlebotomus perniciosus is the most widespread vector of Leishmania infantum in Spain. Laboratory colonisation represents the most feasible source of information on the biology of these insects, but in conducting any study, the density of individuals in the colony may drop to such an extent that it is sometimes difficult to recover the initial population levels. A new technique was tested for the recovery of sandfly eggs in three different colonies; the recovery rate was studied by comparing the standard method of mass rearing with this new method of colony management. The results demonstrate a mean increase of 18.4% in adult production, a growth in colony productivity that justifies the inclusion of this process in the routine maintenance of any colony of sandflies.

The in vivo antimalarial activity of methylene blue combined with pyrimethamine, chloroquine and quinine

The effectiveness of methylene blue (MB) combined with pyrimethamine (PYR), chloroquine (CQ) or quinine (Q) was examined in a classical four-day suppressive test against a causative agent of rodent malaria, Plasmodium berghei. A marked potentiation was observed when MB was administered at a non-curative dose of 15 mg/kg/day in combination with PYR (0.19 mg/kg/day) or Q (25 mg/kg/day). No synergy was found between MB (15 mg/Kg) and CQ (0.75 mg/Kg). Our results suggest that the combination of MB with PYR or Q may improve the efficacy of these currently used antimalarial drugs.

Two new species and new records of biting midges of the genus Culicoides from northwestern Argentina (Diptera: Ceratopogonidae)

The following two new species of Culicoides from the Argentinean Yungas are described, illustrated and placed to subgenus or species group and compared with related congeners: Culicoides calchaqui Spinelli & Veggiani Aybar and Culicoides willinki Spinelli & Veggiani Aybar. Culicoides daedaloides Wirth & Blanton is recorded for the first time for Argentina and Culicoides pseudoheliconiae Felippe-Bauer is firstly mentioned from the northwestern region of the country.

Updated distribution of Aedes albopictus (Diptera: Culicidae) in Spain: new findings in the mainland Spanish Levante, 2013

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia.

Vulnerabilidade da criança diante de situações adversas ao seu desenvolvimento: proposta de matriz analítica

Estudo teórico que teve como objetivo elaborar uma matriz conceitual para analisar a vulnerabilidade da criança diante de situações adversas para seu desenvolvimento. Propõe-se que vulnerabilidade da criança seja analisada por meio dos aspectos das dimensões individual, relativa a relações sustentadoras contínuas e proteção física e segurança; social , concernente à inserção social da família e ao acesso a direitos à proteção e promoção social, e programática , cenário político-programático, de diretrizes e implementação político-programáticas. A aplicação prática desta matriz permite a captação do processo saúde-doença para além da dimensão individual, possibilitando a articulação das políticas públicas e das ações dos profissionais, a fim de obter eficácia no atendimento às necessidades das crianças. O uso desse instrumento conceitual pode propiciar às equipes de saúde a apreensão, de forma específica, das situações adversas ao desenvolvimento infantil, bem como subsidiar a construção de planos de intervenção a partir das dimensões analíticas da vulnerabilidade.

Observations on the biology and distribution of Uresiphita reversalis (Lepidoptera, Crambidae), a defoliator of the native tree Calia secundiflora in México

Observations on the biology and distribution of Uresiphita reversalis (Lepidoptera, Crambidae), a defoliator of the native tree Calia secundiflora in México. Uresiphita reversalis (Guenée, 1854) feeding on Calia secundiflora (Ortega) Yakovlev is recorded for the first time in the state of Hidalgo, Mexico. New aspects regarding the life cycle, feeding behaviour, geographical distribution and host plant damage by U. reversalis on C. secundiflora are here presented and discussed.

Frações de carbono e nitrogênio em função da textura, do relevo e do uso do solo na microbacia do agreste em Vaca Brava (PB)

A produtividade em sistemas agrícolas de subsistência ou de baixos insumos depende do fornecimento de nutrientes oriundos da mineralização da matéria orgânica do solo (MOS). Portanto, a quantidade e a qualidade da MOS são duas variáveis fundamentais relacionadas com a sustentabilidade da produção agropecuária de subsistência. O objetivo do presente trabalho foi analisar as inter-relações de usos do solo, as posições no relevo e suas texturas com os teores totais de C (Ct) e N (Nt), a fração de C oxidável por KMnO4 16,5 mol L-1 (Cox) e a fração de N aminoaçúcar (N amino). Esses atributos foram quantificados em 260 amostras simples de solo (0-20 cm) retiradas da microbacia Vaca Brava (PB), em áreas de uso agropecuário, num sistema de amostragem com arranjo fatorial de estratos. Os teores de Ct, Nt e N amino decresceram na seqüência: pastagem > capineira > roçado. Em relação ao relevo, tanto os teores de Ct e Nt quanto suas frações apresentaram os maiores teores nas áreas de várzea, siltosas e os menores nas áreas de rampa pedimentada, mais arenosas. A redistribuição de sedimentos nas vertentes foi ocasionada por processos erosivos da camada superficial das encostas, nas quais predominam texturas finas. Em relação às classes texturais, os teores de MOS e de suas frações foram maiores (p < 0,05) nas amostras de textura argilo-arenosa e menores nas de textura areia franca. Em todas as situações analisadas, os teores de Cox e N amino tenderam a acompanhar as variações nos teores de Ct e Nt. Por conta desta última tendência, os índices Cox/Ct e Nt/N amino, que representam a qualidade da MOS, indicaram poucas mudanças significativas nesse atributo de acordo com o uso, relevo ou classe textural.

Soil phosphorus fractions in sandy soils amended with cattle manure for long periods

Phosphorus fractions were determined in soil samples from areas fertilized or not with farmyard cattle manure (FYM) and in samples of FYM used in the semi-arid region of Paraiba state, Brazil. Soil samples were taken from the 0-20; 20-40 and 40-60 cm layers of 18 cultivated areas, which, according to interviews with farmers, had been treated with 12 to 20 t ha-1 FYM annually, for the past 2 to 40 years. Soil samples were also collected from four unfertilized pasture areas as controls. Phosphorus in the soil samples was sequentially extracted with water (Pw), resin (Pres), NaHCO3 (Pi bic and Po bic), NaOH (Pi hid and Po hid), H2SO4 (Pacid) and, finally, by digestion with H2SO4/H2O2 (Presd). Nine FYM samples were extracted with water, resin, Mehlich-1, H2SO4, NaOH or digestion with H2SO4/H2O2, not sequentially, and the extracts analyzed for P. The sampled areas had homogeneous, sandy and P-deficient soils; increases in total soil P (Pt) above the mean value of the control areas (up to 274 mg kg-1 in the 0-20 cm layer of the most P-enriched samples) were therefore attributed to FYM applications, which was the only external P input in the region. Regression analysis was used to study the relationship between soil P fractions and Pt. The Pacid fraction, related to Ca-P forms, showed the greatest increases (p < 0.01) as a result of FYM applications, rising from 8.4 mg kg-1 in a non-fertilized sample to 43.8 mg kg-1 in the sample with the highest Pt content. The sum of Pw, Pres and Pi bic, considered as labile P, showed comparable increases with Pacid, while Pi hid showed the smallest increase due to FYM applications. Organic P forms also increased, more so the fraction Po hid, considered less labile, than the more labile one, Po bic. The residual P fraction was practically half of Pt, independently of the Pt value. Increases in labile P, Pacid and organic P were justified by the high average concentration of Pw (36 %), Pacid (34 %), and Po hid (30 %) in the FYM. Significant changes in the proportion of P forms among soil layers indicated the downward movement of P in organic forms.

Eficiência de adubação de batatinha com esterco e N mineral (15N) e efeito residual no cultivo do milheto

Os solos arenosos utilizados para o cultivo da batatinha (Solanum tuberosum) no agreste da Paraíba têm baixo teor de N. Adubações anuais com esterco e N mineral, realizadas empiricamente, têm resultado na acumulação e,ou, perda de nutrientes do solo. Por esse motivo, objetivou-se determinar a eficiência do sulfato de amônio (15N) mais esterco na adubação da batatinha e do efeito residual dessa adubação na produção e absorção de N pelo milheto (Pennisetum glaucum) cultivado em sequência. Em experimento de campo em Neossolo regolítico, foi comparada a combinação de 16 t ha-1 de esterco + 80 kg ha-1 de N, utilizada mais frequentemente na região, com doses de 11 t ha-1 de esterco combinadas com 0, 40 e 80 kg ha-1 de N em aplicação única ou parcelada. O sulfato de amônio utilizado foi enriquecido em 15N (2,5 % de abundância). Em todos os tratamentos, avaliou-se a produtividade de matéria seca de tubérculos e parte aérea do milheto, assim como a composição isotópica do N nesses materiais. Demonstrou-se que a adubação tradicional é excessiva nesse solo, uma vez que a redução para 11 t ha-1 de esterco e 40 kg ha-1 de N mineral não resultou em queda de produtividade das culturas; no entanto, houve queda com a aplicação isolada de esterco. O milheto recuperou grande parte do N mineral adicionado à batatinha e mostrou ser uma alternativa como forrageira em condições de semiárido.