The Three-dimensional Solution Structure of NaD1, a New Floral Defensin from Nicotiana alata and its Application to a Homology Model of the Crop Defense Protein alfAFP

NMR spectroscopy and simulated annealing calculations have been used to determine the three-dimensional structure of NaD1, a novel antifungal and insecticidal protein isolated from the flowers of Nicotiana alata. NaD1 is a basic, cysteine-rich protein of 47 residues and is the first example of a plant defensin from flowers to be characterized structurally. Its three-dimensional structure consists of an a-helix and a triple-stranded anti-parallel beta-sheet that are stabilized by four intramolecular disulfide bonds. NaD1 features all the characteristics of the cysteine-stabilized up motif that has been described for a variety of proteins of differing functions ranging from antibacterial insect defensins and ion channel-perturbing scorpion toxins to an elicitor of the sweet taste response. The protein is biologically active against insect pests, which makes it a potential candidate for use in crop protection. NaD1 shares 31% sequence identity with alfAFP, an antifungal protein from alfalfa that confers resistance to a fungal pathogen in transgenic potatoes. The structure of NaD1 was used to obtain a homology model of alfAFP, since NaD1 has the highest level of sequence identity with alfAFP of any structurally characterized antifungal defensin. The structures of NaD1 and alfAFP were used in conjunction with structure - activity data for the radish defensin Rs-AFP2 to provide an insight into structure-function relationships. In particular, a putative effector site was identified in the structure of NaD1 and in the corresponding homology model of alfAFP. (C) 2002 Elsevier Science Ltd. All rights reserved.

Rapid genotyping of loci involved in antifolate drug resistance in Plasmodium falciparum by primer extension

Current methods used to genotype point mutations in Plasmodium falciparum genes involved in resistance to antifolate drugs include restriction digestion of PCR products, allele-specific amplification or sequencing. Here we demonstrate that known point mutations in dihydrofolate reductase and dihydropteroate synthase can be scored quickly and accurately by single-nucleotide primer extension and detection of florescent products on a capillary sequencer. We use this method to genotype parasites in natural infections from the Thai-Myanmar border. This approach could greatly simplify large-scale screening of resistance mutations of the type required for evaluating and updating antimalarial drug treatment policies. The method can be easily adapted to other P. falciparum genes and will greatly simplify scoring of point mutations in this and other parasitic organisms. © 2002 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.

Effect of sorghum resistant to Stenodiplosis sorghicola (Coquillett) (Diptera: Cecidomyiidae) on oviposition and development of Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae)

Helicoverpa armigera oviposition preference for, and larval development on sorghum hybrids with differing resistance to sorghum midge, Stenodiplosis sorghicola, were investigated. When H. armigera larvae were fed seed of resistant and susceptible hybrids in the laboratory there were no differences in larval and pupal sizes or the rate of development. The same result was recorded when larvae fed on panicles on plants in a glasshouse. On some sampling occasions, significantly more eggs were laid on panicles of resistant hybrids in the field. This occurred when plants were in plots and also in a mixed planting. Midge-resistance status did not affect levels of egg parasitism. In a field study using recombinant inbred lines between a midge-resistant and a midge-susceptible line, no relationship was found between level of resistance and oviposition of H. armigera. We conclude that, although midge-resistant hybrids are sometimes preferred for oviposition by H. armigera, the resistance per se does not determine this preference. Egg survival, larval survival, development and resultant damage are not significantly affected by the midge-resistance status of the host.

Pathogenic specialisation within Colletotrichum trifolii in Australia, and lucerne cultivar reactions to all known Australian pathotypes

Anthracnose and crown rot, caused by Colletotrichum trifolii, are serious diseases of lucerne (Medicago saliva L.) in humid regions of the world. A race survey was conducted by inoculating individual lucerne clones (genotypes) with C. trifolii isolates collected from a range of Medicago hosts, locations, and years in south-eastern Queensland. This survey revealed for the first time in Australia the presence of race 2 (virulence on anthracnose resistance gene An I) and the first world report of race 4 (virulence on An(2)). A collection of North American race I and race 2 C. trifolii isolates, when inoculated onto the Australian differential clones, gave responses that were in agreement with their North American reactions. A RAPD analysis was conducted on 9 Australian C. trifolii isolates including races 1, 2, and 4; two C. destructivum and one C. gloeosporioides isolate were included as known outliers. For the C. trifolii isolates, 94.6% similarity was found regardless of host origin or race, compared with 2.2% similarity between this group and the C. gloeosporioides and C. destructivum isolates, confirming that the new races belong to C. trifolii. Currently, it is hypothesised that only plants carrying genes An, and An2 are resistant to the 3 races. Of 22 cultivars screened against the 3 races, only UQL-1, Hallmark, and Pioneer 54Q53 had >30% of plants resistant to the 3 races in separate screenings. The research highlights the need to find new sources of resistance to C. trifolii in lucerne.

Increased expression of the SNARE accessory protein Munc18c in lipid-mediated insulin resistance

Fatty acids inhibit insulin-mediated glucose metabolism in skeletal muscle, an effect largely attributed to defects in insulin-mediated glucose transport. Insulin-resistant mice transgenic for the overexpression of lipoprotein lipase (LPL) in skeletal muscle were used to examine the molecular mechanism(s) in more detail. Using DNA gene chip array technology, and confirmation by RT-PCR and Western analysis, increases in the yeast Sec1p homolog Munc18c mRNA and protein were found in the gastrocnemius muscle of transgenic mice, but not other tissues. Munc18c has been previously demonstrated to impair insulin-mediated glucose transport in mammalian cells in vitro. Of interest, stably transfected C2C12 cells overexpressing LPL not only demonstrated increases in Munc18c mRNA and protein but also in transcription rates of the Munc18c gene. jlr To confirm the relevance of fatty acid metabolism and insulin resistance to the expression of Munc18c in vivo, a 2-fold increase in Munc18c protein was demonstrated in mice fed a high-fat diet for 4 weeks. Together, these data are the first to implicate in vivo increases in Munc18c as a potential contributing mechanism to fatty acid-induced insulin resistance.

Anomalous water absorption in porous materials

The absorption of fluid by unsaturated, rigid porous materials may be characterized by the sorptivity. This is a simple parameter to determine and is increasingly being used as a measure of a material's resistance to exposure to fluids (especially moisture and reactive solutes) in aggressive environments. The complete isothermal absorption process is described by a nonlinear diffusion equation, with the hydraulic diffusivity being a strongly nonlinear function of the degree of saturation of the material. This diffusivity can be estimated from the sorptivity test. In a typical test the cumulative absorption is proportional to the square root of time. However, a number of researchers have observed deviation from this behaviour when the infiltrating fluid is water and there is some potential for chemo-mechanical interaction with the material. In that case the current interpretation of the test and estimation of the hydraulic diffusivity is no longer appropriate. Kuntz and Lavallee (2001) discuss the anomalous behaviour and propose a non-Darcian model as a more appropriate physical description. We present an alternative Darcian explanation and theory that retrieves the earlier advantages of the simple sorptivity test in providing parametric information about the material's hydraulic properties and allowing simple predictive formulae for the wetting profile to be generated.

Prediction of genetic gain from selection indices for disease resistance in papaya hybrids

In order to select superior hybrids for the concentration of favorable alleles for resistance to papaya black spot, powdery mildew and phoma spot, 67 hybrids were evaluated in two seasons, in 2007, in a randomized block design with two replications. Genetic gains were estimated from the selection indices of Smith & Hazel, Pesek & Baker, Williams, Mulamba & Mock, with selection intensity of 22.39%, corresponding to 15 hybrids. The index of Mulamba & Mock showed gains more suitable for the five traits assessed when it was used the criterion of economic weight tentatively assigned. Together, severity of black spot on leaves and on fruits, characteristics considered most relevant to the selection of resistant materials, expressed percentage gain of -44.15%. In addition, there were gains for other characteristics, with negative predicted selective percentage gain. The results showed that the index of Mulamba & Mock is the most efficient procedure for simultaneous selection of papaya hybrid resistant to black spot, powdery mildew and phoma spot.

Rootstocks resistant to Meloidogyne incognita and compatibility of grafting in net melon

Due to the few studies about grafting in net melon, in order to obtain better control of soil pathogens, the aim of the present study was to evaluate 16 genotypes of Cucurbitaceae: Benincasa hispida, Luffa cylindrica, pumpkin 'Jacarezinho', pumpkin 'Menina Brasileira', squash 'Exposição', squash 'Coroa', pumpkin 'Canhão Seca', pumpkin 'Squash', pumpkin 'Enrrugado Verde', pumpkin 'Mini Paulista', pumpkin 'Goianinha', watermelon 'Charleston Gray', melon 'Rendondo Gaucho', melon 'Redondo Amarelo', cucumber 'Caipira HS' and cucumber 'Caipira Rubi', regarding to compatibility of grafting in net melon and resistance to Meloidogyne incognita, based on the reproduction factor (RF), according to Oostenbrink (1966). To assess resistance, the seedlings were transplanted to ceramic pots and inoculated with 300/mL eggs and/or second stage juveniles of M. incognita. At 50 days after transplanting, the plants were removed from the pots and the resistance was evaluated. The compatibility between resistant rootstock and grafts of net melon was determined by performing simple cleft grafting, in a commercial net melon hybrid of great market acceptance and susceptible to M. incognita (Bonus no. 2). The genotypes Luffa cylindrica, pumpkin 'Goianinha', pumpkin 'Mini-Paulista', melon 'Redondo Amarelo', watermelon 'Charleston Gray' are resistant to the nematode M. incognita. The better compatibilities occurred with the rootstocks melon 'Amarelo', which presented 100% of success, followed by pumpkin 'Mini-Paulista' with 94%. On the other hand, Sponge gourd, watermelon 'Charleston Gray' and pumpkin 'Goianinha' showed low graft take percentages of 66%, 62% and 50%, respectively.

Agronomic performance of 27 cocoa progenies and plant selection based on productivity, self-compatibility and disease resistance

The objective of the present work was to evaluate 27 progenies of cocoa crosses considering the agronomic traits and select F1 plants within superior crosses. The experiment was installed in March 2005, in the Experimental Station Joaquim Bahiana (ESJOB), in Itajuipe, Bahia. The area of the experiment is of approximately 3 ha, with a total of 3240 plants. Thirteen evaluations of vegetative brooms, five of cushion brooms and 15 of number of pods per plant were accomplished. Thirty pollinations were made for each selected plant to test for self-compatibility. The production, based on the number of pods per plant, and resistance to witches´ broom indicated CEPEC 94 x CCN 10, RB 39 x CCN 51 and CCN 10 x VB 1151 as superior progenies. All selections tested were self-compatible. The analyses of progenies and individual tree data, associated to visual field observations, allowed the selection of 17 plants which were included in a network of regional tests to determine the phenotypic stability.

From antimicrobial agents to health promotion

A emergência de multiresistência apresentada por microrganismos é um dos grandes desafios que enfrentam actualmente os profissionais de Saúde e a população em geral. Os factores que contribuem para o desenvolvimento de resistência a antibióticos na comunidade podem ser categorizados como comportamentais ou ambientais/políticas. O objectivo deste trabalho foi caracterizar a situação actual na visão dos Pais de alunos do pré-escolar e 1º ciclo. De modo a avaliar as necessidades de intervenção e as actividades a serem desenvolvidas, um instrumento para estudar os hábitos e comportamentos adoptados na utilização de antibióticos, foi adaptado, validado e aplicado numa amostra piloto.

Yeast resistance and its reversion

Actualmente, a resistência a drogas é um dos grandes problemas na terapia de diferentes patologias e também na biotecnologia e agricultura. O objectivo do presente estudo foi elucidar a resistência a drogas em leveduras patogénicas ou alimentares, que mais frequentemente são encontradas. Assim, diferentes antifungicos clínicos e novos compostos recentemente sintetizados foram testados por um método para avaliar a resistência e sua reversão que foi previamente desenvolvido. Foi testado um painel de diferentes drogas para reverter a resistência. Mostramos que alguns destes moduladores foram capazes de reverter eficazmente a resistência.

ß-Lactams: chemical structure, mode of action and mechanisms of resistance

This synopsis summarizes the key chemical and bacteriological characteristics of β-lactams, penicillins, cephalosporins, carbanpenems, monobactams and others. Particular notice is given to first-generation to fifth-generation cephalosporins. This review also summarizes the main resistance mechanism to antibiotics, focusing particular attention to those conferring resistance to broad-spectrum cephalosporins by means of production of emerging cephalosporinases (extended-spectrum β-lactamases and AmpC β-lactamases), target alteration (penicillin-binding proteins from methicillin-resistant Staphylococcus aureus) and membrane transporters that pump β-lactams out of the bacterial cell.

β-Lactams: chemical structure, mode of action and mechanisms of resistance

This synopsis summarizes the key chemical and bacteriological characteristics of β-lactams, penicillins, cephalosporins, carbanpenems, monobactams and others. Particular notice is given to first-generation to fifth-generation cephalosporins. This reviewalso summarizes the main resistancemechanism to antibiotics, focusing particular attention to those conferring resistance to broad-spectrum cephalosporins by means of production of emerging cephalosporinases (extended-spectrum β-lactamases and AmpC β-lactamases), target alteration (penicillin-binding proteins from methicillin-resistant Staphylococcus aureus) and membrane transporters that pump β-lactams out of the bacterial cell.

Performance of concrete made with aggregates recycled from precasting industry waste: influence of the crushing process

The aim of this paper is to evaluate the influence of the crushing process used to obtain recycled concrete aggregates on the performance of concrete made with those aggregates. Two crushing methods were considered: primary crushing, using a jaw crusher, and primary plus secondary crushing (PSC), using a jaw crusher followed by a hammer mill. Besides natural aggregates (NA), these two processes were also used to crush three types of concrete made in laboratory (L20, L45 e L65) and three more others from the precast industry (P20, P45 e P65). The coarse natural aggregates were totally replaced by coarse recycled concrete aggregates. The recycled aggregates concrete mixes were compared with reference concrete mixes made using only NA, and the following properties related to the mechanical and durability performance were tested: compressive strength; splitting tensile strength; modulus of elasticity; carbonation resistance; chloride penetration resistance; water absorption by capillarity; water absorption by immersion; and shrinkage. The results show that the PSC process leads to better performances, especially in the durability properties. © 2014 RILEM

Variation in response of Schistosoma mansoni strains to schistosomicides

The BH strain of Schistosoma mansoni was found to be highly susceptible, to hy cant hone (1 x 80 mg/kg), oxamniquine (1 x 100 mg/kg) niridazole (5 x 100 mg/kg), praziquantel (1 x 100 mg/kg), oltipraz (5 x 125 mg/kg) and amoscanate (1 x 300 mg/kg) and is therefore a good reference strain for chemotherapeutic trials. By contrast, the MPR-1 strain was shown to have developed resistance to oxamniquine without ever having been dosed with oxamniquine. Other oxamniquine/hycanthone resistant strains were shown to have maintained their resistance and a strain believed to be partially resistant to oltipraz was evaluated.