Mortality of Plutella xylostella larvae treated with Aspidosperma pyrifolium ethanol extracts

The objective of this work was to assess the effects of Aspidosperma pyrifolium ethanol extracts on cabbage moth (Plutella xylostella) larvae. The ethanol extracts of the stem bark, fruits and roots of A. pyrifolium were obtained by classical phytochemical methods, and the resulting subfractions were tested on P. xylostella, using 4 and 5 mg L-1. The crude ethanol extract of the stem bark was more lethal. The alkaloid-rich aqueous subfraction derived from the stem bark extract caused 100% larval mortality at 4 mg L-1. Insecticidal activity was associated with the presence of the monoterpenoid indole alkaloids aspidofractinine, 15-demethoxypyrifoline, and N-formylaspidofractinine. These alkaloids presented excellent insecticidal properties against P. xylostella.

Egg parasitoid wasps as natural enemies of the neotropical stink bug Dichelops melacanthus

The objective of this work was to determine the potential of five species of Scelionidae wasps - Telenomus podisi, Trissolcus basalis, Trissolcus urichi, Trissolcus teretis and Trissolcus brochymenae - as natural enemies of the neotropical stink bug Dichelops melacanthus, and to determine if the presence of eggs of other stink bug species influences the parasitism and development of the parasitoids. Two kinds of experiments were done in laboratory: without choice of hosts (eggs of D. melacanthus) and with choice (eggs of D. melacanthus and of Euschistus heros). Biological parameters, including proportion of parasitism, immature survivorship, progeny sex ratio, immature stage development period, and host preference were recorded. All the evaluated parasitoids can parasitize and develop on D. melacanthus eggs. The first choice of eggs did not influence the proportion of D. melacanthus eggs parasitized by Tr. basalis, Tr. teretis or Tr. brochymenae. However, D. melacanthus eggs as the first choice of Te. podisi and Tr. urichi increased, respectively, 9 and 14 times the chance for parasitism on eggs of this species. Behavioral and ecological aspects of parasitoids should be considered prior to their use in biological control programs.

Comparison of the organization and fine structure of a chicken and a Xenopus laevis vitellogenin gene.

The structural organization and the coding nucleotide sequence of the Xenopus laevis A2 and the chicken major vitellogenin genes have been compared. Both genes show the same exon-intron organization. However, the degree of homology between the nucleotide and derived amino acid sequences varies extensively along the genes. Several of the 35 exons are quite similar, and a unique cysteine motif in the lipovitellin II domain is conserved between the two genes. In contrast, one internal region is quite divergent. Part of this region encodes phosvitin, which appears to have evolved rapidly by both point mutations and duplications of serines or short other amino acid stretches. On the basis of these observations, we discuss the possible mechanism of evolution of phosvitin in vertebrates.

Structure of the extracellular domains of human and Xenopus Fn14: implications in the evolution of TWEAK and Fn14 interactions.

TWEAK (TNF homologue with weak apoptosis-inducing activity) and Fn14 (fibroblast growth factor-inducible protein 14) are members of the tumor necrosis factor (TNF) ligand and receptor super-families. Having observed that Xenopus Fn14 cross-reacts with human TWEAK, despite its relatively low sequence homology to human Fn14, we examined the conservation in tertiary fold and binding interfaces between the two species. Our results, combining NMR solution structure determination, binding assays, extensive site-directed mutagenesis and molecular modeling, reveal that, in addition to the known and previously characterized β-hairpin motif, the helix-loop-helix motif makes an essential contribution to the receptor/ligand binding interface. We further discuss the insight provided by the structural analyses regarding how the cysteine-rich domains of the TNF receptor super-family may have evolved over time. DATABASE: Structural data are available in the Protein Data Bank/BioMagResBank databases under the accession codes 2KMZ, 2KN0 and 2KN1 and 17237, 17247 and 17252. STRUCTURED DIGITAL ABSTRACT: TWEAK binds to hFn14 by surface plasmon resonance (View interaction) xeFn14 binds to TWEAK by enzyme linked immunosorbent assay (View interaction) TWEAK binds to xeFn14 by surface plasmon resonance (View interaction) hFn14 binds to TWEAK by enzyme linked immunosorbent assay (View interaction).

Functional and morphological evidence for a ventricular conduction system in zebrafish and Xenopus hearts.

Zebrafish and Xenopus have become popular model organisms for studying vertebrate development of many organ systems, including the heart. However, it is not clear whether the single ventricular hearts of these species possess any equivalent of the specialized ventricular conduction system found in higher vertebrates. Isolated hearts of adult zebrafish (Danio rerio) and African toads (Xenopus laevis) were stained with voltage-sensitive dye and optically mapped in spontaneous and paced rhythms followed by histological examination focusing on myocardial continuity between the atrium and the ventricle. Spread of the excitation wave through the atria was uniform with average activation times of 20 +/- 2 and 50 +/- 2 ms for zebrafish and Xenopus toads, respectively. After a delay of 47 +/- 8 and 414 +/- 16 ms, the ventricle became activated first in the apical region. Ectopic ventricular activation was propagated significantly more slowly (total ventricular activation times: 24 +/- 3 vs. 14 +/- 2 ms in zebrafish and 74 +/- 14 vs. 35 +/- 9 ms in Xenopus). Although we did not observe any histologically defined tracts of specialized conduction cells within the ventricle, there were trabecular bands with prominent polysialic acid-neural cell adhesion molecule staining forming direct myocardial continuity between the atrioventricular canal and the apex of the ventricle; i.e., the site of the epicardial breakthrough. We thus conclude that these hearts are able to achieve the apex-to-base ventricular activation pattern observed in higher vertebrates in the apparent absence of differentiated conduction fascicles, suggesting that the ventricular trabeculae serve as a functional equivalent of the His-Purkinje system.

Loss of egg yolk genes in mammals and the origin of lactation and placentation

Embryonic development in nonmammalian vertebrates depends entirely on nutritional reserves that are predominantly derived from vitellogenin proteins and stored in egg yolk. Mammals have evolved new resources, such as lactation and placentation, to nourish their developing and early offspring. However, the evolutionary timing and molecular events associated with this major phenotypic transition are not known. By means of sensitive comparative genomics analyses and evolutionary simulations, we here show that the three ancestral vitellogenin-encoding genes were progressively lost during mammalian evolution (until around 30-70 million years ago, Mya) in all but the egg-laying monotremes, which have retained a functional vitellogenin gene. Our analyses also provide evidence that the major milk resource genes, caseins, which have similar functional properties as vitellogenins, appeared in the common mammalian ancestor approximately 200-310 Mya. Together, our data are compatible with the hypothesis that the emergence of lactation in the common mammalian ancestor and the development of placentation in eutherian and marsupial mammals allowed for the gradual loss of yolk-dependent nourishment during mammalian evolution

Changes in activity of the regulatory glycolytic enzymes and of the pyruvate-dehydrogenase complex during the development of Xenopus laevis.

In this study we investigated the variations of the maximal activities of the rate-controlling glycolytic enzymes (i.e., hexokinase, HK; phosphofructokinase, PFK; pyruvate kinase, PK) and of the pyruvate-dehydrogenase complex (PDHc) during the early embryogenesis of Xenopus laevis (from cleavage through hatching). All the enzymatic assays, using different coupled reactions, were performed spectrophotometrically on cytosolic and mitochondrial fractions. The maximal HK activity increases markedly from neurulation onwards, PFK activity presents a peak around gastrulation, PK activity remains relatively constant throughout the period studied and the highest PDHc activity is observed during cleavage. The specific activities display the same temporal pattern. Furthermore, in the sequence of reactions by which glucose is degraded to form acetyl-CoA, the maximal activities of PFK and PK are not limiting while those of HK and PDHc could be rate-limiting at relatively late developmental stages (hatching).

Recognition of insect eggs in Arabidopsis modulates defense responses

Plants activate direct and indirect defenses in response to insect egg deposition. In Arabidopsis thaliana, oviposition by the butterfly Pieris brassicae triggers cellular and molecular changes that are similar to the changes caused by biotrophic pathogens. Even though this innate immune response did not affect egg survival in Arabidopsis, we could show that different insect eggs elicit specific gene expression changes. Additionally, egg- induced necrosis could be observed in a variety of plants from different families ranging from dicotyledonous plants to monocots, suggesting that insect-egg detection by plants is a widespread mechanism and that different insect species contain elicitors of immune responses. Extracts from caterpillars and eggs contain elicitors that co-purified over several extraction steps. Chemical fractionation of caterpillar extracts lead to the characterisation of an active compound that was determined to be a triglyceride by NMR analysis. The exact structure of the side chains as well as the elicitor's presence in insect eggs have yet to be confirmed.We also found that the plant defense signal salicylic acid (SA) accumulates at the site of oviposition. This is unexpected, as the SA pathway controls the defense against fungal and bacterial pathogens whereas it negatively interacts with the jasmonic acid (JA) pathway, which is crucial for the defense against herbivores. Application of P. brassicae or Spodoptera littoralis egg extract onto leaves reduced the induction of insect-responsive genes after challenge with caterpillars, suggesting that egg-derived elicitors suppress plant defense. Consequently, larval growth of the generalist herbivore S. littoralis, but not of the specialist P. brassicae, was significantly higher on plants treated with egg extract than on control plants. In contrast, suppression of gene induction and enhanced S. littoralis performance were not found in the SA-deficient mutant sid2-l, indicating that SA mediates this phenomenon. These data reveal an intriguing facet of the crosstalk between SA- and JA-signalling pathways and suggest that insects have evolved a way to suppress the induction of defense genes by laying eggs that release elicitors. Additionally, we demonstrated that mutants of known crosstalk regulators, including nprl-1, tga2356, ein2-l and wrky70-l, are not affected in egg-induced suppression of herbivore defenses. JA treatment was not able to alleviate this SA/JA negative crosstalk, suggesting that this suppression operates through a novel mechanism downstream of JA biosynthesis.

Bioactivity of Piper extracts on Tuta absoluta (Lepidoptera: Gelechiidae) in tomato

The objective of this work was to evaluate the bioactivity of ethanolic leaf extracts from four species of the genus Piper against the tomato leafminer Tuta absoluta (Lepidoptera: Gelechiidae) under laboratory conditions. The evaluated species were: P. amalago var. medium, P. glabratum, P. mikanianum, and P. mollicomum. In the initial screening assay (extract concentration of 2,000 mg L-1), all tested extracts caused significant larval mortality, particularly the extract of P. amalago var. medium; however, no extracts reduced the weight of the surviving larvae. The extract from P. amalago var. medium at the concentration of 1,011 mg L-1 caused a significant lengthening of the larval and pupal stages. The ethanolic leaf extract of P. amalago var. medium is promising for the control of T. absoluta larvae in tomato, since it exhibits acute toxicity toward these caterpillars at the concentration of 2,000 mg L-1 and affects the insect's development by reducing its survival and lengthening the larval and pupal stages.

Limestone particle sizes and lighting regimens on egg and bone quality of laying hens

Abstract:The objective of this work was to evaluate the effect of limestone particle sizes in the diet and of lighting regimes on the egg and bone quality and on the performance of commercial laying hens. Three hundred Hissex White layers, at 18 weeks of age, were distributed in a completely randomized design, in a 5×2 factorial arrangement (coarse limestone in the diet at 0, 25, 50, 75, and 100%; with or without artificial light), with five replicates of six birds. No significant interaction was observed between particle sizes and lighting regime for the evaluated parameters. There was no significant effect of coarse limestone level in the diet on the performance and egg quality of hens; however, bone deformity (3.23 to 4.01 mm), strength (5.19 to 6.70 kgf cm-2), and mineral matter (51.09 to 59.61%) improved as the proportion of coarse limestone increased. For lighting regime, the treatment with artificial light yielded higher Haugh unit values (87.17 vs. 85.54) than that with natural light only. Greater limestone particles improve bone quality of laying hens, and the use of artificial light can benefit the albumen quality of the eggs.

Screening for wound-induced oxylipins in Arabidopsis thaliana by differential HPLC-APCI/MS profiling of crude leaf extracts and subsequent characterisation by capillary-scale NMR

A simple non-targeted differential HPLC-APCI/MS approach has been developed in order to survey metabolome modifications that occur in the leaves of Arabidopsis thaliana following wound-induced stress. The wound-induced accumulation of metabolites, particularly oxylipins, was evaluated by HPLC-MS analysis of crude leaf extracts. A generic, rapid and reproducible pressure liquid extraction procedure was developed for the analysis of restricted leaf samples without the need for specific sample preparation. The presence of various oxylipins was determined by head-to-head comparison of the HPLC-MS data, filtered with a component detection algorithm, and automatically compared with the aid of software searching for small differences in similar HPLC-MS profiles. Repeatability was verified in several specimens belonging to different series. Wound-inducible jasmonates were efficiently highlighted by this non-targeted approach without the need for complex sample preparation as is the case for the 'oxylipin signature' procedure based on GC-MS. Furthermore this HPLC-MS screening technique allowed the isolation of induced compounds for further characterisation by capillary-scale NMR (CapNMR) after HPLC scale-up. In this paper, the screening method is described and applied to illustrate its potential for monitoring polar and non-polar stress-induced constituents as well as its use in combination with CapNMR for the structural assignment of wound-induced compounds of interest

Translation and assembly of HLA-DR antigens in Xenopus oocytes injected with mRNA from a human B-cell line.

HLA-DR antigens are polymorphic cell surface glycoproteins, expressed primarily in B lymphocytes and macrophages, which are thought to play an important role in the immune response. Two polypeptide chains, alpha and beta, are associated at the cell surface, and a third chain associates with alpha and beta intracellularly. RNA isolated from the human B-cell line Raji was injected in Xenopus laevis oocytes. Immunoprecipitates of translation products with several monoclonal antibodies revealed the presence of HLA-DR antigens similar to those synthesized in Raji cells. One monoclonal antibody was able to bind the beta chain after dissociation of the three polypeptide chains with detergent. The presence of all three chains was confirmed by two-dimensional gel electrophoresis. The glycosylation pattern of the three chains was identical to that observed in vivo, as evidenced in studies using tunicamycin, an inhibitor of N-linked glycosylation. The presence of alpha chains assembled with beta chains in equimolar ratio was further demonstrated by amino-terminal sequencing. An RNA fraction enriched for the three mRNAs, encoding alpha, beta, and intracellular chains, was isolated. This translation-assembly system and the availability of monoclonal antibodies make it possible to assay for mRNA encoding specific molecules among the multiple human Ia-like antigens.

The avocado fruit borer, Stenoma catenifer (wals.) (Lepidoptera: elachistidae): egg and damage distribution and parasitism

The avocado fruit borer, Stenoma catenifer (Wals.) has been a limiting factor in growing avocados over the last years in many Brazilian states. This is a result of the lack of safe and feasible management practices to minimize the fruit borer damage. The aim of this study was to obtain information on the pest biology and ecology as well as on the role of natural enemies to define strategies to control the pest. Samples were taken biweekly and consisted of 20 fruits collected randomly (10 from the upper half and 10 from the lower half of the plant) in ten plants, cv. Margarida, in a commercial avocado grove in Arapongas and Cambé regions, PR, from October/2001 to September/2002. Laboratory determinations of the percentage of damaged fruit per plant region, location and number of bored fruit sites, and the number and location of the fruit borer eggs, including parasitized ones, were performed. The results showed that S. catenifer preferred to oviposit and attack fruits located on the upper half of the trees. The majority of the eggs were laid on the fruit pedicel whereas the damage was mainly located on the lower half of the fruits. Trichogrammatids were the most constant and abundant parasitoids found in both localities throughout the study period.

When cholesterol is not cholesterol: a note on the enzymatic determination of its concentration in model systems containing vegetable extracts

Background: Experimental evidences demonstrate that vegetable derived extracts inhibit cholesterol absorption in the gastrointestinal tract. To further explore the mechanisms behind, we modeled duodenal contents with several vegetable extracts. Results: By employing a widely used cholesterol quantification method based on a cholesterol oxidase-peroxidase coupled reaction we analyzed the effects on cholesterol partition. Evidenced interferences were analyzed by studying specific and unspecific inhibitors of cholesterol oxidase-peroxidase coupled reaction. Cholesterol was also quantified by LC/MS. We found a significant interference of diverse (cocoa and tea-derived) extracts over this method. The interference was strongly dependent on model matrix: while as in phosphate buffered saline, the development of unspecific fluorescence was inhibitable by catalase (but not by heat denaturation), suggesting vegetable extract derived H2O2 production, in bile-containing model systems, this interference also comprised cholesterol-oxidase inhibition. Several strategies, such as cholesterol standard addition and use of suitable blanks containing vegetable extracts were tested. When those failed, the use of a mass-spectrometry based chromatographic assay allowed quantification of cholesterol in models of duodenal contents in the presence of vegetable extracts. Conclusions: We propose that the use of cholesterol-oxidase and/or peroxidase based systems for cholesterol analyses in foodstuffs should be accurately monitored, as important interferences in all the components of the enzymatic chain were evident. The use of adequate controls, standard addition and finally, chromatographic analyses solve these issues.