Fungicidal synergism of fluconazole and cyclosporine in Candida albicans is not dependent on multidrug efflux transporters encoded by the CDR1, CDR2, CaMDR1, and FLU1 genes.

The combination of fluconazole (FLC) and cyclosporine (CY) is fungicidal in FLC-susceptible C. albicans (O. Marchetti, P. Moreillon, M. P. Glauser, J. Bille, and D. Sanglard, Antimicrob. Agents Chemother. 44:2373-2381, 2000). The mechanism of this synergism is unknown. CY has several cellular targets including multidrug efflux transporters. The hypothesis that CY might inhibit FLC efflux was investigated by comparing the effect of FLC-CY in FLC-susceptible parent CAF2-1 (FLC MIC, 0.25 mg/liter) and in FLC-hypersusceptible mutant DSY1024 (FLC MIC, 0.03 mg/liter), in which the CDR1, CDR2, CaMDR1, and FLU1 transporter genes have been selectively deleted. We postulated that a loss of the fungicidal effect of FLC-CY in DSY1024 would confirm the roles of these efflux pumps. Time-kill curve studies showed a more potent fungistatic effect of FLC (P = 0.05 at 48 h with an inoculum of 10(3) CFU/ml) and a more rapid fungicidal effect of FLC-CY (P = 0.05 at 24 h with an inoculum of 10(3) CFU/ml) in the FLC-hypersusceptible mutant compared to those in the parent. Rats with experimental endocarditis were treated for 2 or 5 days with high-dose FLC, high-dose CY, or both drugs combined. FLC monotherapy for 5 days was more effective against the hypersusceptible mutant than against the parent. However, the addition of CY to FLC still conferred a therapeutic advantage in animals infected with mutant DSY1024, as indicated by better survival (P = 0.04 versus the results obtained with FLC) and sterilization of valves and kidneys after a very short (2-day) treatment (P = 0.009 and 0.002, respectively, versus the results obtained with FLC). Both in vitro and in vivo experiments consistently showed that the deletion of the four membrane transporters in DSY1024 did not result in loss of the fungicidal effect of FLC-CY. Yet, the accelerated killing in the mutant suggested a "dual-hit" mechanism involving FLC hypersusceptibility due to the efflux pump elimination and fungicidal activity conferred by CY. Thus, inhibition of multidrug efflux transporters encoded by CDR1, CDR2, CaMDR1, and FLU1 genes is not responsible for the fungicidal synergism of FLC-CY. Other cellular targets must be considered.

Molecular features of hepatosplenic T-cell lymphoma unravels potential novel therapeutic targets.

The pathogenesis of hepatosplenic T-cell lymphoma (HSTL), a rare entity mostly derived from γδ T cells and usually with a fatal outcome, remains largely unknown. In this study, HSTL samples (7γδ and 2αβ) and the DERL2 HSTL cell line were subjected to combined gene-expression profiling and array-based comparative genomic hybridization. Compared with other T-cell lymphomas, HSTL had a distinct molecular signature irrespective of TCR cell lineage. Compared with peripheral T-cell lymphoma, not otherwise specified and normal γδ T cells, HSTL overexpressed genes encoding NK-cell-associated molecules, oncogenes (FOS and VAV3), the sphingosine-1-phosphatase receptor 5 involved in cell trafficking, and the tyrosine kinase SYK, whereas the tumor-suppressor gene AIM1 (absent in melanoma 1) was among the most down-expressed. We found highly methylated CpG islands of AIM1 in DERL2 cells, and decitabine treatment induced a significant increase in AIM1 transcripts. Syk was present in HSTL cells and DERL2 cells contained phosphorylated Syk and were sensitive to a Syk inhibitor in vitro. Genomic profiles confirmed recurrent isochromosome 7q (n = 6/9) without alterations at the SYK and AIM1 loci. Our results identify a distinct molecular signature for HSTL and highlight oncogenic pathways that offer rationale for exploring new therapeutic options such as Syk inhibitors and demethylating agents.

Intracellular trafficking of interleukin-1 receptor I requires Tollip.

Interleukin-1 receptor (IL-1RI) is a master regulator of inflammation and innate immunity. When triggered by IL-1beta, IL-1RI aggregates with IL-1R-associated protein (IL-1RAcP) and forms a membrane proximal signalosome that potently activates downstream signaling cascades. IL-1beta also rapidly triggers endocytosis of IL-1RI. Although internalization of IL-1RI significantly impacts signaling, very little is known about trafficking of IL-1RI and therefore about precisely how endocytosis modulates the overall cellular response to IL-1beta. Upon internalization, activated receptors are often sorted through endosomes and delivered to lysosomes for degradation. This is a highly regulated process that requires ubiquitination of cargo proteins as well as protein-sorting complexes that specifically recognize ubiquitinated cargo. Here, we show that IL-1beta induces ubiquitination of IL-1RI and that via these attached ubiquitin groups, IL-1RI interacts with the ubiquitin-binding protein Tollip. By using an assay to follow trafficking of IL-1RI from the cell surface to late endosomes and lysosomes, we demonstrate that Tollip is required for sorting of IL-1RI at late endosomes. In Tollip-deficient cells and cells expressing only mutated Tollip (incapable of binding IL-1RI and ubiquitin), IL-1RI accumulates on late endosomes and is not efficiently degraded. Furthermore, we show that IL-1RI interacts with Tom1, an ubiquitin-, clathrin-, and Tollip-binding protein, and that Tom1 knockdown also results in the accumulation of IL-1RI at late endosomes. Our findings suggest that Tollip functions as an endosomal adaptor linking IL-1RI, via Tom1, to the endosomal degradation machinery.

Rapid [Gamma]-amino acid synthesis and the inhibition of the growth and development of oblique banded leaf-roller larvae

The hypothesis that rapid y-aminobutyric acid (GABA) accumulation is a plant defense against phytophagous insects was investigated. Simulation of mechanical damage resulting from phytophagous insect activity increased soybean (Glycine max L.) leaf GABA 10- to 25-fold within 1 to 4 min. Pulverizing leaf tissue resulted in a value of 2. 15 (±O. 11 SE) ~mol GABA per gram fresh weight. Increasing the GABA levels in a synthetic diet from 1.6 to 2.6 Jlffiol GABA per gram fresh weight reduced the growth rates, developmental rates, total biomass (50% reduction), and survival rates (30% reduction) of cultured Oblique banded leaf-roller (OBLR) (Choristonellra rosacealla Harris) larvae. In field experiments OBLR larvae were found predominantly on young terminal leaves which have a reduced capacity to produce GABA in response to mechanical damage. Glutamate decarboxylase (GAD) is a cytosolic enzyme which catalyses the decarboxylation of L-Glu to GABA. GAD is a calmodulin binding enzyme whose activity is stimulated dramatically by increased cytosolic H+ or Ca2 + ion concentrations. Phytophagous insect activity will disrupt the cellular compartmentation of H+ and Ca2 +, activate GAD and subsequent GABA accumulation. In animals GABA is a major inhibitory neurotransmitter. The possible mechanisms resulting in GABA inhibited growth and development of insects are discussed.

The identification, distribution and persistence of oxamyl and its degradation products in planted corn seed, seedling root and soil from oxamyl-treated corn seeds

A simple method was developed for treating corn seeds with oxamyl. It involved soaking the seeds to ensure oxamyl uptake, centrifugation to draw off excess solution, and drying under a stream of air to prevent the formation of fungus. The seeds were found to have an even distribution of oxamyl. Seeds remained fungus-free even 12 months after treatment. The highest nonphytotoxic treatment level was obtained by using a 4.00 mg/mL oxamyl solution. Extraction methods for the determination of oxamyl (methyl-N'N'-dimethyl-N-[(methylcarbamoyl)oxy]-l-thiooxamimidate), its oxime (methyl-N',N'-dimethyl-N-hydroxy-1-thiooxamimidate), and DMCF (N,N-dimethyl-1-cyanoformanade) in seed" root, and soil were developed. Seeds were processed by homogenizing, then shaking in methanol. Significantly more oxamyl was extracted from hydrated seeds as opposed to dry seeds. Soils were extracted by tumbling in methanol; recoveries range~ from 86 - 87% for oxamyl. Root was extracted to 93% efficiency for oxamyl by homogenizing the tissue in methanol. NucharAttaclay column cleanup afforded suitable extracts for analysis by RP-HPLC on a C18 column and UV detection at 254 nm. In the degradation study, oxamyl was found to dissipate from the seed down into the soil. It was also detected in the root. Oxime was detected in both the seed and soil, but not in the root. DMCF was detected in small amounts only in the seed.

Évaluation de l'acquisition de la résistance à la colistine chez Escherichia coli O149 chez le porc.

La diarrhée post-sevrage est une maladie d’importance dans l’industrie porcine et est principalement causée Escherichia coli O149. Le traitement habituellement utilisé est la néomycine. Cependant, en raison de l’antibiorésistance, les vétérinaires se tournent vers la colistine sulfate (CS). La CS lie les lipopolysaccharides (LPS) et provoque un déplacement des cations divalents causant la formation de pores entrainant la mort cellulaire. Le système à deux composantes PmrA/PmrB est le plus incriminé dans la résistance à la colistine en ajoutant un groupement 4-amino-4-déoxy-L-arabinose (L-Ara4N) au lipide A des LPS, augmentant ainsi la charge du LPS et diminuant son affinité pour la CS. L’objectif principal est d’évaluer l’acquisition de la résistance à la CS d’E. coli in vitro et dans un modèle in vivo. Nous avons utilisé des souches associées à des cas cliniques d’E. coli O149 et avons créé 22 mutants résistants à la CS. La concentration minimale inhibitrice (CMI) a été mesurée par une méthode de double dilution et comparée au seuil de résistance. Suite au séquençage des gènes pmrA/pmrB, nous avons identifié sept nouveaux polymorphismes, trois dans PmrA : A80V, N128I, S144G et quatre dans PmrB : V87E, D148Y, D148V et T156M. Pour l’essai in vivo, nous avons suivi une souche expérimentale ETEC:F4 (E. coli O149) et isolé des E. coli de la flore commensale. Le séquençage des gènes pmrA et pmrB de ces isolats a montré un polymorphisme spécifique, G15R et T156M respectivement. Cependant, plusieurs souches récoltées possédaient une résistance à la CS, mais sans polymorphisme de PmrA/PmrB, suggérant d’autre(s) mécanisme(s) de résistance.

Bt-cotton boosts the gross margin of small-scale cotton producers in South Africa

This paper explores some of the issues involved in the Genetic Modification (GM) debate by focusing on one crop that has been modified for pest resistance, cotton (Gossypium hirsutum), and commercially released to small-scale farmers in the Makhathini Flats, KwaZulu Natal, the Republic of South Africa. This was the first commercial release of a GM variety (Bt-cotton) in Sub-Saharan Africa, and thus provides valuable and timely insights into some of the potential advantages and disadvantages of the technology for small-scale farmers in Africa. Even though there are wider concerns regarding the vulnerability of small-scale farmers in the area, the survey results suggest that Bt-cotton generated higher yields and gross margins than non-Bt-cotton. In addition, Bt-cotton significantly reduced the use of pesticide with consequent potential benefits to human health and the environment.

Multilocus Analyses of Seven Candidate Genes Suggest Interacting Pathways for Obesity-Related Traits in Brazilian Populations

We investigated whether variants in major candidate genes for food intake and body weight regulation contribute to obesity-related traits under a multilocus perspective. We studied 375 Brazilian subjects from partially isolated African-derived populations (quilombos). Seven variants displaying conflicting results in previous reports and supposedly implicated in the susceptibility of obesity-related phenotypes were investigated: beta(2)-adrenergic receptor (ADRB2) (Arg16Gly), insulin induced gene 2 (INSIG2) (rs7566605), leptin (LEP) (A19G), LEP receptor (LEPR) (Gln223Arg), perilipin (PLIN) (6209T > C), peroxisome proliferator-activated receptor-gamma (PPARG) (Pro12Ala), and resistin (RETN) (-420C > G). Regression models as well as generalized multifactor dimensionality reduction (GMDR) were employed to test the contribution of individual effects and higher-order interactions to BMI and waist-hip ratio (WHR) variation and risk of overweight/obesity. The best multilocus association signal identified in the quilombos was further examined in an independent sample of 334 Brazilian subjects of European ancestry. In quilombos, only the PPARG polymorphism displayed significant individual effects (WHR variation, P = 0.028). No association was observed either with the risk of overweight/obesity (BMI >= 25 kg/m(2)), risk of obesity alone (BMI >= 30 kg/m(2)) or BMI variation. However, GMDR analyses revealed an interaction between the LEPR and ADRB2 polymorphisms (P = 0.009) as well as a third-order effect involving the latter two variants plus INSIG2 (P = 0.034) with overweight/obesity. Assessment of the LEPR-ADRB2 interaction in the second sample indicated a marginally significant association (P = 0.0724), which was further verified to be limited to men (P = 0.0118). Together, our findings suggest evidence for a two-locus interaction between the LEPR Gln223Arg and ADRB2 Arg16Gly variants in the risk of overweight/obesity, and highlight further the importance of multilocus effects in the genetic component of obesity.

Herança da resistência ao enfezamento em milho

Os objetivos deste trabalho foram determinar a herança da resistência ao complexo de enfezamento em milho e determinar as melhores fontes de resistência entre as linhagens estudadas. Foram realizadas as análises dialélica e médias de gerações em linhagens de milho. Para a análise dialélica, foram cruzadas 12 linhagens de milho, em dialélico parcial. Para análises de médias de gerações, foram cruzadas três linhagens resistentes e quatro suscetíveis, para a obtenção das gerações F1, F2, RCP R e RCP S. Os trabalhos foram conduzidos em Jaboticabal, SP. A incidência de enfezamento foi avaliada no estádio fenológico R3. Efeitos significativos quanto à capacidade geral de combinação e capacidade específica de combinação foram obtidos, o que indicou que, no controle do caráter enfezamentos, estão envolvidos tanto os efeitos aditivos quanto os de dominância. Análises de médias de gerações mostraram a presença de poucos genes envolvidos com o controle da resistência, com predominância de efeitos aditivos, o que permite a seleção de genótipos resistentes. As linhagens L02, L03 e L05 poderão ser utilizadas como fontes de resistência, em futuras combinações híbridas.

Efeito da alimentação de Schizaphis graminum com genótipos de sorgo no desenvolvimento do predador Cycloneda sanguinea

O pulgão Schizaphis graminum (Rondani) (Hemiptera: Aphididae) é uma das principais pragas do sorgo. O objetivo deste trabalho foi determinar o efeito da alimentação deste inseto com genótipos de sorgo resistentes e suscetíveis no desenvolvimento do predador Cycloneda sanguinea (Linnaeus) (Coleoptera: Coccinellidae). O pulgão foi criado em folhas dos genótipos de sorgo GR 11111 e TX 430 x GR 111, resistentes, GB 3B, de resistência moderada e BR 007B, suscetível ao hemíptero. As larvas de C. sanguinea foram alimentadas diariamente, ad libitum, com ninfas e adultos de S. graminum. A duração das fases de desenvolvimento e a sobrevivência de C. sanguinea não foram influenciadas pelo genótipo. Apenas a duração do quarto ínstar, da fase larval e do período de larva a adulto foram afetadas. O peso de larvas de segundo ínstar de C. sanguinea não foi afetado pelo genótipo. Nos demais ínstares e fase adulta, esse parâmetro foi diferenciado conforme o genótipo de sorgo utilizado como hospedeiro ao pulgão. Os genótipos resistentes GR 11111 e TX 430 x GR 111 não afetam adversamente o desenvolvimento e a fecundidade de C. sanguinea, durante uma geração. Estes genótipos demonstram compatibilidade com o predador e viabilizam o manejo de S. graminum na cultura do sorgo.

Virulence factors of Escherichia coli isolated from diarrheic calves

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Suscetibilidade de genótipos de feijão ao vírus-do-mosaico-dourado

O objetivo deste trabalho foi avaliar a suscetibilidade de diversos genótipos de feijão ao vírus-do-mosaico-dourado (VMDF), transmitido pela mosca branca (Bemisia tabaci). A semeadura foi realizada na época da seca e das águas, com e sem aplicação do inseticida granulado Aldicarb (3,0 kg ha-1 do i.a.) no sulco de semeadura. O delineamento experimental utilizado foi o de blocos casualizados disposto em esquema fatorial 14x2, representado por genótipos e inseticida, respectivamente, com quatro repetições. A maior infestação de mosca-branca e incidência do vírus ocorreu na época da seca, causando prejuízos à produção do feijoeiro. Os genótipos apresentaram diferentes graus de suscetibilidade ao vírus e ao inseto vetor. Os genótipos mais tolerantes foram IAPAR 57, IAPAR 65, IAPAR 72, Ônix, Aporé e 606 (5)(214-17). A aplicação do inseticida sistêmico controla o vetor em ambas as épocas de cultivo, proporcionando aumentos da produtividade.

Novo registro de Trichospilus diatraeae Cherian & Margabandhu, 1942 (Hymenoptera: Eulophidae), como parasitóide de Spodoptera cosmioides Walker, 1858 (Lepidoptera: Noctuidae) no Brasil

Spodoptera cosmioides (Lepidoptera: Noctuidae)é uma espécie polífaga e alimenta-se de grande número de plantas cultivadas e espontâneas. No Brasil, as culturas do abacaxi, algodoeiro, arroz, berinjela, cebola, eucalipto, pimentão e tomateiro, entre outras hortaliças, são consideradas hospedeiras da praga. Entretanto, apesar da ampla gama de hospedeiros, sua ocorrência como praga é relatada relacionada a desequilíbrios provocados pelo uso excessivo de inseticidas de amplo espectro, o que vem causando a resistência da praga a inseticidas. Devido a essa complexidade, métodos alternativos de controle têm sido propostos, por exemplo, o controle biológico através da utilização de parasitóides. Trichospilus diatraeae (Hymenptera: Eulophidae) é um parasitóide gregário pupal preferencialmente de espécies da ordem Lepidoptera. Este é o primeiro relato de T. diatraeae parasitando pupas de S. cosmioides, parasitóide que oferece novas perspectivas para os programas de controle biológico.

Resposta de cultivares de arroz de sequeiro ao preparo do solo e à irrigação por aspersão

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de diferentes intervalos de recuperação entre as séries sobre o desempenho muscular no exercício leg-press em idosas não treinadas

OBJETIVO: Verificar a influência de dois diferentes intervalos de recuperação (IR) entre as séries no exercício leg-press sobre o número e sustentabilidade das repetições e no volume total, em idosas não treinadas. MÉTODOS: Onze idosas (66,5 ± 5,0 anos; 59,2 ± 9,1kg; 146,4 ± 34,9cm) foram submetidas a duas sessões experimentais de exercícios com pesos com intensidade de 15 repetições máximas. Cada sessão experimental foi composta por três séries realizadas até a fadiga muscular utilizando IR de um (IR-1) ou três minutos (IR-3). As sessões experimentais foram separadas por, no mínimo, 48 horas. Todas as participantes realizaram ambos os protocolos e um delineamento cross-over balanceado foi utilizado para determinar a ordem das sessões experimentais. RESULTADOS: Para ambos os IR entre as séries, reduções significativas (P < 0,05) no número e na sustentabilidade das repetições foram observadas da primeira para a segunda e terceira séries e da segunda para a terceira séries. Diferenças significativas (P < 0,05) entre os IR foram observadas nas duas séries finais. O volume total da sessão realizada com IR-3 foi estatisticamente superior (20,4%; P < 0,05) quando comparada a sessão IR-1. CONCLUSÃO: O número e a sustentabilidade das repetições e o volume total de treino de idosas não treinadas são influenciados pelo IR empregado entre as séries. Maiores IR devem ser utilizados quando a finalidade for otimizar o volume de treino por meio da sustentabilidade das repetições. Em contrapartida, menores IR devem ser utilizados quando a meta for obter maiores níveis de fadiga muscular.