Antibodies against the Plasmodium falciparum glutamate-rich protein from naturally exposed individuals living in a Brazilian malaria-endemic area can inhibit in vitro parasite growth

The glutamate-rich protein (GLURP) is an exoantigen expressed in all stages of the Plasmodium falciparum life cycle in humans. Anti-GLURP antibodies can inhibit parasite growth in the presence of monocytes via antibody-dependent cellular inhibition (ADCI), and a major parasite-inhibitory region has been found in the N-terminal R0 region of the protein. Herein, we describe the antiplasmodial activity of anti-GLURP antibodies present in the sera from individuals naturally exposed to malaria in a Brazilian malaria-endemic area. The anti-R0 antibodies showed a potent inhibitory effect on the growth of P. falciparum in vitro, both in the presence (ADCI) and absence (GI) of monocytes. The inhibitory effect on parasite growth was comparable to the effect of IgGs purified from pooled sera from hyperimmune African individuals. Interestingly, in the ADCI test, higher levels of tumour necrosis factor alpha (TNF-α) were observed in the supernatant from cultures with higher parasitemias. Our data suggest that the antibody response induced by GLURP-R0 in naturally exposed individuals may have an important role in controlling parasitemia because these antibodies are able to inhibit the in vitro growth of P. falciparum with or without the cooperation from monocytes. Our results also indicate that TNF-α may not be relevant for the inhibitory effect on P. falciparum in vitro growth.

Hepatitis C virus quantification in serum and saliva of HCV-infected patients

The hepatitis C virus (HCV) can be detected in blood and other bodily fluids, such as saliva, semen and gastric juices. The aim of this study was to compare the HCV viral loads in the serum and saliva of infected patients. Twenty-nine patients with detectable HCV RNA in their serum and saliva were included in this study. The HCV viral loads were determined through quantitative real-time polymerase chain reactions. The median viral RNA levels were 5.78 log10 copies in the serum and 3.32 log10 copies in the saliva. We observed that the salivary HCV viral load was significantly lower than the viral load in the serum. Further studies are required to understand the role of saliva in the diagnosis, management and potential transmission of HCV.

The zinc finger protein TcZFP2 binds target mRNAs enriched during Trypanosoma cruzi metacyclogenesis

Trypanosomes are parasitic protozoa in which gene expression is primarily controlled through the regulation of mRNA stability and translation. This post-transcriptional control is mediated by various families of RNA-binding proteins, including those with zinc finger CCCH motifs. CCCH zinc finger proteins have been shown to be essential to differentiation events in trypanosomatid parasites. Here, we functionally characterise TcZFP2 as a predicted post-transcriptional regulator of differentiation in Trypanosoma cruzi. This protein was detected in cell culture-derived amastigotes and trypomastigotes, but it was present in smaller amounts in metacyclic trypomastigote forms of T. cruzi. We use an optimised recombinant RNA immunopreciptation followed by microarray analysis assay to identify TcZFP2 target mRNAs. We further demonstrate that TcZFP2 binds an A-rich sequence in which the adenosine residue repeats are essential for high-affinity recognition. An analysis of the expression profiles of the genes encoding the TcZFP2-associated mRNAs throughout the parasite life cycle by microarray hybridisation showed that most of the associated mRNAs were upregulated in the metacyclic trypomastigote forms, also suggesting a role for TcZFP2 in metacyclic trypomastigote differentiation. Knockdown of the orthologous Trypanosoma brucei protein levels showed ZFP2 to be a positive regulator of specific target mRNA abundance.

Fibronectin modulates thymocyte-thymic epithelial cell interactions following Trypanosoma cruzi infection

Developing thymocytes interact with thymic epithelial cells (TECs) through cell-cell interactions, TEC-derived secretory moieties and extracellular matrix (ECM)-mediated interactions. These physiological interactions are crucial for normal thymocyte differentiation, but can be disrupted in pathological situations. Indeed, there is severe thymic atrophy in animals acutely infected with Trypanosoma cruzi due to CD4+CD8+ thymocyte depletion secondary to caspase-mediated apoptosis, together with changes in ECM deposition and thymocyte migration. We studied an in vitro model of TEC infection by T. cruzi and found that infected TEC cultures show a reduced number of cells, which was likely associated with decreased proliferative capacity, but not with increased cell death, as demonstrated by bromodeoxyuridine and annexin-V labelling. The infected TEC cultures exhibited increased expression of fibronectin (FN), laminin (LM) and type IV collagen. Importantly, treatment with FN increased the relative number of infected cells, whereas treatment with anti-FN or anti-LM antibodies resulted in lower infection rates. Consistent with these data, we observed increased thymocyte adhesion to infected TEC cultures. Overall, these results suggest that ECM molecules, particularly FN, facilitate infection of the thymic epithelium and that the consequent enhancement of ECM expression might be associated with changes in TEC-thymocyte interactions.

Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

Influence of age on the haemoglobin concentration of malaria-infected patients in a reference centre in the Brazilian Amazon

Anaemia is amongst the major complications of malaria, a major public health problem in the Amazon Region in Latin America. We examined the haemoglobin (Hb) concentrations of malaria-infected patients and compared it to that of malaria-negative febrile patients and afebrile controls. The haematological parameters of febrile patients who had a thick-blood-smear performed at an infectious diseases reference centre of the Brazilian Amazon between December 2009-January 2012 were retrieved together with clinical data. An afebrile community control group was composed from a survey performed in a malaria-endemic area. Hb concentrations and anaemia prevalence were analysed according to clinical-epidemiological status and demographic characteristics. In total, 7,831 observations were included. Patients with Plasmodium falciparum infection had lower mean Hb concentrations (10.5 g/dL) followed by P. vivax-infected individuals (12.4 g/dL), community controls (12.8 g/dL) and malaria-negative febrile patients (13.1 g/dL) (p < 0.001). Age, gender and clinical-epidemiological status were strong independent predictors for both outcomes. Amongst malaria-infected individuals, women in the reproductive age had considerably lower Hb concentrations. In this moderate transmission intensity setting, both vivax and falciparum malaria are associated with reduced Hb concentrations and risk of anaemia throughout a wide age range.

BAY 41-2272 activates host defence against local and disseminated Candida albicans infections

In our previous study, we have found that 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]-pyrimidin-4-ylamine (BAY 41-2272), a guanylate cyclase agonist, activates human monocytes and the THP-1 cell line to produce the superoxide anion, increasing in vitro microbicidal activity, suggesting that this drug can be used to modulate immune functioning in primary immunodeficiency patients. In the present work, we investigated the potential of the in vivo administration of BAY 41-2272 for the treatment of Candida albicans and Staphylococcus aureus infections introduced via intraperitoneal and subcutaneous inoculation. We found that intraperitoneal treatment with BAY 41-2272 markedly increased macrophage-dependent cell influx to the peritoneum in addition to macrophage functions, such as spreading, zymosan particle phagocytosis and nitric oxide and phorbol myristate acetate-stimulated hydrogen peroxide production. Treatment with BAY 41-2272 was highly effective in reducing the death rate due to intraperitoneal inoculation of C. albicans, but not S. aureus. However, we found that in vitro stimulation of peritoneal macrophages with BAY 41-2272 markedly increased microbicidal activities against both pathogens. Our results show that the prevention of death by the treatment of C. albicans-infected mice with BAY 41-2272 might occur primarily by the modulation of the host immune response through macrophage activation.

Molecular characterisation of the NSP4 gene of group A human rotavirus G2P[4] strains circulating in São Paulo, Brazil, from 1994 and 2006 to 2010

Group A human rotaviruses (HuRVA) are causative agents of acute gastroenteritis. Six viral structural proteins (VPs) and six nonstructural proteins (NSPs) are produced in RV-infected cells. NSP4 is a diarrhoea-inducing viral enterotoxin and NSP4 gene analysis revealed at least 15 (E1-E15) genotypes. This study analysed the NSP4 genetic diversity of HuRVA G2P[4] strains collected in the state of São Paulo (SP) from 1994 and 2006-2010 using reverse transcription-polymerase chain reaction, sequencing and phylogenetic analysis. Forty (97.6%) G2P[4] strains displayed genotype E2; one strain (2.4%) displayed genotype E1. These results are consistent with the proposed linkage between VP4/VP7 (G2P[4]) and the NSP4 (E2) genotype of HuRVA. NSP4 phylogenetic analysis showed distinct clusters, with grouping of most strains by their genotype and collection year, and most strains from SP were clustered together with strains from other Brazilian states. A deduced amino acid sequence alignment for E2 showed many variations in the C-terminal region, including the VP4-binding domain. Considering the ability of NSP4 to generate host immunity, monitoring NSP4 variations, along with those in the VP4 or VP7 protein, is important for evaluating the circulation and pathogenesis of RV. Finally, the presence of one G2P[4]E1 strain reinforces the idea that new genotype combinations emerge through reassortment and independent segregation.

Determinantes do desenvolvimento do setor agropecuário nos municípios

Dada a importância do setor agropecuário para a economia brasileira, é fundamental conhecer os fatores determinantes do desenvolvimento agropecuário em seus municípios. Sob a luz de referencial teórico que aborda questões relacionadas aos fatores determinantes do desenvolvimento agropecuário e do desenvolvimento econômico local, neste estudo fez-se uso das metodologias de análise multivariada conhecidas como análise fatorial e análise de cluster. Utilizou-se o estado de Minas Gerais como recorte analítico. Foram selecionadas 22 variáveis para cada município, representando diferentes dimensões do desenvolvimento, visando verificar quais os fatores determinantes do desenvolvimento agropecuário. Após a análise fatorial, optou-se pela extração de seis fatores com raiz característica maior do que um e que respondem, em conjunto, por 62,25% da variância total dos dados: desenvolvimento econômico, investimento público, qualidade de vida, condições da atividade agropecuária, condições de meio ambiente e consumo, e condições de financiamento. Pela análise de cluster, foram criados cinco grupos, de acordo com o desempenho dos membros nos fatores. Os resultados corroboram os apontamentos literários em quase sua totalidade e, além disso, destaca-se e discute-se a existência de um ciclo positivo gerado pelo incremento dos fatores determinantes do desenvolvimento agropecuário e pelas vantagens competitivas. Neste estudo, aponta-se a importância do investimento público na promoção do desenvolvimento, reforçando o proposto por trabalhos anteriores de que o Estado desempenha papel fundamental para garantir condições que propiciem o desenvolvimento do setor agropecuário. Outra constatação factível é a de que fatores isolados, tais como as boas condições da agricultura e do meio ambiente, não são capazes de propiciar desenvolvimento para o setor nos municípios mineiros.

From print to screen: changes and challenges facing the Brazilian publishing industry

The publishing industry is at a turning point. Facing the first major disruptive innovation in five centuries, its long-established structure and business model are at stake. Building on literature based on the pitfalls for incumbents, we interviewed key executives from the major publishers in Brazil to understand their perspective. We find that not only are they facing those pitfalls, but we also propose a new one, The Industry View Trap, concerning challenges created by convergence, the difficulty to deal with changes in the ecosystem and the fact that the very definition of the industry you're part of might have changed.

Testes em condições para o controle de Dysmicoccus texensis (Tinsley) (Hemiptera, Pseudococcidae) em cafeeiro com nematóides entomopatogênicos do gênero Heterorhabditis(Rhabditida, Heterorhabditidae)

Os nematóides entomopatogênicos (NEPs) apresentam potencial para o controle biológico de pragas e têm sido usados com sucesso, em vários países, no controle de pragas de solo e de ambientes crípticos, como a cochonilha-da-raiz-do-cafeeiro Dysmicoccus texensis (Tinsley). Testes de laboratório demonstram que estes agentes apresentam alta virulência sobre este inseto, no entanto, são necessários testes que avaliem a eficiência dos NEPs em condições de casa-de-vegetação e campo, sendo este o objetivo do presente trabalho. O experimento em condição de casa-de-vegetação para o controle da cochonilha foi realizado em vasos infestados, usando dois isolados do nematóide e dois métodos de aplicação (cadáver infectado e suspensão aquosa), conduzido em delineamento inteiramente casualisado com cinco repetições. O experimento em condições de campo foi conduzido em blocos casualisados (6 blocos), para avaliar a eficiência de dois isolados heterorhabditídeos no controle da cochonilha-da-raiz-do-cafeeiro. Os resultados mostraram que, em casa-de-vegetação, o método de suspensão aquosa apresentou melhores resultados para os dois isolados, sendo que JPM3 aplicado em suspensão aquosa foi o melhor tratamento, apresentando eficiência de controle de 70%. No experimento de campo, apenas o tratamento com o inseticida Actara 250 WG (thiamethoxam), usado como padrão de comparação, e JPM3, aplicado em suspensão aquosa, diferiram da testemunha, apresentando 81 e 65% de eficiência de controle, respectivamente.

The aquatic habit and host plants of Paracles klagesi (Rothschild) (Lepidoptera, Erebidae, Arctiinae) in Brazil

The aquatic habit and host plants of Paracles klagesi (Rothschild) (Lepidoptera, Erebidae, Arctiinae) in Brazil. The aquatic caterpillar Paracles klagesi (Rothschild, 1910) was collected from the headwaters of a stream in an ecotone between Cerrado and Babaçu forest in northeastern Brazil. The single caterpillar found was observed feeding on the macrophyte Tonina fluviatilis Aubl. (Eriocaulaceae) and other aquatic plants of the family Nymphaeaceae present in the area, but also accepted as food Elodea canadensis Michx. (Hydrocharitaceae) and Cabomba sp. (Cabombaceae) under laboratory conditions.

Carbono orgânico, nitrogênio total, biomassa e atividade microbiana do solo em duas cronosseqüências de reabilitação após a mineração de bauxita

A mineração é uma das atividades antrópicas que causa grande impacto nos ecossistemas e o grau de degradação depende da intensidade de interferência no solo, do volume explorado e do rejeito produzido. Os microrganismos do solo e sua atividade, por participarem de funções-chave, podem fornecer indicações sobre o real estado de reabilitação de áreas mineradas. O objetivo deste trabalho foi avaliar o impacto da atividade de mineração em duas cronosseqüências com diferentes estratégias (revegetação) de reabilitação em solos de área de mineração de bauxita sobre diversos atributos: C de compostos orgânicos (CO), N total do solo (Nt), biomassa microbiana, atividade enzimática e respiração do solo. O estudo foi desenvolvido em áreas pertencentes a Alcoa S/A, em dois ambientes distintos: (a) em áreas com vegetação típica de campo, predominantemente herbácea, em altitude inferior a 1.000 m, e (b) em áreas de montanhas, no topo do planalto, incluindo remanescentes de cobertura vegetal nativa representada por floresta subtropical subcaducifólia, a uma altitude de 1.600 m, denominadas serra, com características diferenciadas quanto à estratégia e ao tempo de reabilitação, variando de áreas recém-mineradas à áreas com 19 anos de reabilitação. Coletaram-se amostras compostas com três repetições em oito áreas no campo e nove na serra, em duas profundidades (0-10 e 10-20 cm), no inverno e verão. A mineração de bauxita provocou grande impacto nos atributos estudados, causando reduções nos seus valores de até 99 %. Tanto no campo como na serra, os teores de CO, de Nt, a biomassa microbiana e a atividade enzimática aumentaram com as diferentes estratégias de reabilitação. Entretanto a resposta dos atributos estudados foi diferenciada em função do tempo de reabilitação. A biomassa microbiana e a atividade enzimática apresentaram recuperação muito rápida, atingindo valores semelhantes aos das áreas de referência a partir do primeiro ano, enquanto para o CO e o Nt estes só foram alcançados de modo consistente em períodos mais longos de reabilitação (18 anos). O coeficiente metabólico (qCO2) foi indicativo do estresse provocado pela mineração, mas não diferenciou os diferentes tempos de reabilitação. Os resultados deste estudo mostram que os atributos essenciais ao funcionamento adequado do solo são recuperáveis pela revegetação.

Atributos físicos, químicos e biológicos de solo de cerrado sob diferentes sistemas de uso e manejo

À medida que o conhecimento do sistema plantio direto se amplia, verifica-se que o uso de indicadores químicos isolados não permite melhor caracterização dos solos, sendo necessário utilizar um conjunto de indicadores da qualidade do solo com a entrada de outros atributos, entre eles os físicos e os biológicos. Objetivou-se avaliar os efeitos de sistemas de manejo e uso do solo nos atributos físicos, químicos e biológicos de um Latossolo Vermelho distrófico e um Neossolo Quartzarênico órtico sob Cerrado, no entorno do Parque Nacional das Emas. Os aspectos avaliados no Latossolo foram: Cerrado nativo, pastagem, milheto em preparo convencional, nabo forrageiro em plantio direto e sorgo em plantio direto. No Neossolo: Cerrado nativo, pastagem nativa, integração agricultura-pecuária, pastagem cultivada, plantio direto com soja no verão e plantio direto com milho no verão. As amostras de solo foram coletadas na profundidade de 0 a 10 cm. O delineamento experimental foi o inteiramente casualizado, com cinco parcelas de 150 m², sendo coletadas 10 subamostras aleatórias. As análises químicas, físicas e biológicas foram realizadas no Laboratório de Solos da UFG/CJ. Os manejos promoveram alterações na densidade do solo, volume total de poros, macroporos e resistência do solo à penetração no Neossolo e no Latossolo, excetuando-se neste o volume total de poros. Houve pequena variação nos atributos químicos nos dois solos, com o Cerrado apresentando maior acidez potencial e menor teor de cátions trocáveis e P. Os atributos biológicos do solo foram alterados pelos sistemas de manejo, sendo mais prejudicados em sistemas com maior revolvimento do solo. A análise canônica dos dados demonstrou que os atributos físicos foram os de menor importância por apresentar maior coeficiente de ponderação nas variáveis canônicas. Os atributos do solo, isoladamente, pouco contribuíram para a avaliação da qualidade do solo: no entanto, quando se usou a análise multivariada, subsidiaram a constatação dos manejos do solo mais sustentáveis.

Ammonia volatilization of urea in the out-of-season corn

The aim of this study was to evaluate the N losses due to volatilization at different rates of common urea, polymer coated urea and urease inhibitor-treated urea in the out-of-season corn, using semi-open static collectors. The treatments consisted of N levels on side-dressing fertilization with urea in different treatments: (a) control (without N), (b) urea 40 kg ha-1 N, (c) urea 80 kg ha-1 N, (d) polymer coated urea 40 kg ha-1 N, (e) polymer coated urea 80 kg ha-1 N and (f) urea with the urease inhibitor (UI) N 80 kg ha-1 N. The results showed that the treatments with polymer coated urea and with urease inhibitor-treated urea reduced the volatilization of N around 50 % compared to common urea, either in the first and the second N side-dressing fertilizations. Thus, they demonstrate that the polymer coat and the urease inhibitors were effective in reducing the volatilization of urea N applied in coverage, which resulted in higher productivity. There was also increasing urease activity in the treatments with application of common urea.