A micelle nucleation model for the interaction of dodecyl sulphate with Lys49-phospholipases A(2)

Bothropstoxin-I (BthTx-I) is a Lys49-PLA(2) from the venom of Bothrops jararacussu that lacks detectable catalytic activity, yet causes rapid Ca2+-independent membrane damage. With the aim of understanding the interaction between BthTx-I and amphiphilic molecules, we have studied the interaction of sodium dodecyl sulphate (SDS) with the protein. Circular dichroism and attenuated total reflection Fourier-transform infrared spectra of BthTx-I reveal changes in the alpha-helical organization of the protein at an SDS/BthTx-I molar ratio of 20-25. At SDS/BthTx-I ratios of 40-45 the alpha-helices return to a native-like conformation, although fluorescence emission anisotropy measurements of 2-amino-N-hexadecyl-benzamide (AHBA) demonstrate that the total SDS is below the critical micelle concentration when this transition occurs. These results may be interpreted as the result of SDS accumulation by the BthTx-I homodimer and the formation of a pre-micelle SDS/BthTx-I complex, which may subsequently be released from the protein surface as a free micelle. Similar changes in the alpha-helical organization of BthTx-I were observed in the presence of dipalmitoylphosphatidylcholine liposomes, suggesting that protein structure transitions coupled to organization changes of bound amphiphiles may play a role in the Ca2+-independent membrane damage by Lys49-PLA(2)s. (c) 2006 Elsevier B.V. All rights reserved.

An ultraviolet photoacoustic spectroscopy study of the interaction between Lys49-phospholipase A(2) and amphiphilic molecules

We have used near ultraviolet photoacoustic spectroscopy (PAS) over the wavelength range 240-320 nm to investigate the complex formed between the homodimeric bothropstoxin-I, a lysine-49-phospholipase A(2) from the venom of Bothrops jararacussu (BthTx-I), with the anionic amphiphile sodium dodecyl sulfate (SDS). At molar ratios > 10, the complex developed a significant light scatter, accompanied by a decrease in the intrinsic tryptophan fluorescence intensity emission (ITFE) of the protein, and an increase in the near UV-PAS signal. Difference PAS spectroscopy at SDS/BthTx-I ratios < 8 were limited to the region 280-290 nm, suggesting initial SDS binding to the tryptophan 77 located at the dimer interface. At SDS/BthTx-I ratios > 10, the intensity between 260 and 320 nm increases demonstrating that the more widespread tyrosine and phenylalanine residues contribute to the SDS/BthTx-I interaction. PAS signal phase changes at wavelengths specific for each aromatic residue suggest that the Trp77 becomes more buried on SDS binding, and that protein structural changes and dehydration may alter the microenvironments of Tyr and Phe residues. These results demonstrate the potential of near UV-PAS for the investigation of membrane proteins/detergent complexes in which light scatter is significant. (c) 2006 Elsevier B.V. All rights reserved.

Photoacoustic spectroscopy of aromatic amino acids in proteins

This paper concerns the use of photoacoustic spectroscopy (PAS) to study the presence of aromatic amino acid in proteins. We examined the aromatic amino acids in six proteins with well-known structures using absorption spectra of near ultraviolet PAS over the wavelength range 240-320 nm. The fundamental understanding of the physical and chemical properties that govern the absorption of light and a subsequent release of heat to generate a transient pressure wave was used to test the concept of monitoring aromatic amino acids with this method. Second derivative spectroscopy in the ultraviolet region of proteins was also used to study the regions surrounding the aromatics and the percentage area in each band was related in order to determine the contribution in function of the respective molar extinction coefficients for each residue. Further investigation was conducted into the interaction between sodium dodecyl sulphate (SDS) and bothropstoxin-I (BthTx-I), with the purpose of identifying the aromatics that participate in the interaction. The clear changes in the second derivative and curve-fitting procedures suggest that initial SDS binding to the tryptophan located in the dimer interface and above 10 SDS an increased intensity between 260 and 320 nm, demonstrating that the more widespread tyrosine and phenylalanine residues contribute to the SDS/BthTx-I interactions. These results demonstrate the potential of near UV-PAS for the investigation of membrane proteins/detergent complexes in which light scattering is significant.

Association of the stilt bug Jalysus ossesae Henry (Hemiptera : Heteroptera : Berytidae) with myrmecophytic plants of the genus Maieta (Melastomataceae) in an upland forest area in central Amazon, Brazil

Maieta guianensis Aubl. and M. poeppigii Mart. ex. Triana (Melastomataceae) are among the most common myrmecophytic plants in the Amazonian forest understory. These myrmecophytes are colonized exclusively by the ants Pheidole minutula Mayr or Crematogaster sp. and usually host two other arthropods, the spider Faiditus subflavus Exline and Levi and the recently described stilt bug Jalysus ossesae Henry. In this study, the association between J ossesae and the myrmecophytic plants M. guianensis and M. poeppigii in an upland forest area in central Amazon, Brazil, is described. The presence of the stilt bugs on M. guianensis and M. poeppigii and on plants around these myrmecophytes was recorded in five transects. The number and position of the stilt bugs on the leaf surface (upper or lower) and leaf type (with or without domatia) of these myrmecophytes, as well as their behavioral acts, were recorded. Jalysus ossesae was found only on the myrmecophytic plants M. guianensis and M. poeppigii. The stilt bug occurred at similar frequencies on M. guianensis and M. poeppigii, and the number of leaves significantly influenced the presence and number of stilt bugs on these myrmecophytes. Feeding, agonistic interaction between males, and mating were observed. Our data indicate that J. ossesae uses the myrmecophytes M. guinanensis and M. poeppigii as reproductive and foraging sites.

Burrowing behavior of Dermatonotus muelleri (Anura, Microhylidae) with reference to the origin of the burrowing behavior of Anura

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Flavonoid increase in soybean as a response to Nezara viridula injury and its effect on insect-feeding preference

The ability of the stink bug (Nezara viridula) to induce and/or increase production of chemical defenses, i.e., flavonoids, in immature seeds of five genotypes of soybean (BR-16, IAC-100, PI 227687, PI 229358, and PI 274454) was investigated under greenhouse and laboratory conditions. Samples from pods of each genotype damaged by stink bug were analyzed for flavonoid content with high performance liquid chromatography. A dual-choice test was conducted to evaluate the feeding preference of N. viridula comparing BR-16 pods treated with extracts of PI 227687 seeds (with and without stink-bug injury), with water-treated pods. Seeds of PI 227687 damaged by N. viridula presented the highest concentration (352 mug/g) of daidzin (4'-hydroxyisoflavone-7-glucoside). The same trend was observed with genistin (4',5,7-trihydroxyisoflavone-7-glucoside): PI 227687 contained 142.4 mug/g, PI 274454, 31.6 mug/g, and PI 229358, 38.9 mug/g. Seeds damaged by stink bugs had higher isoflavone contents (daidzin and genistin), compared to controls. However, after being damaged, PI 274454 and PI 229358 produced less genistin than the other genotypes and no differences in concentration between damaged and nondamaged plants of this genotypes were observed. The numbers of observations of the insect feeding and the numbers of stylet sheaths left in water-treated BR-16 pods were greater than in those treated with PI 227687 extracts. The insects fed for longer periods on BR-16 pods treated with extract of PI 227687 without injury compared to those that were treated with extract of PI 227687 previously injured by stink bugs. Extracts of PI 227687 pods ( damaged or not) were deterrent to adults of N. viridula, and insect injury increased concentrations of daidzin and genistin in PI 227687 seeds. The deterrence seemed to be more pronounced after pods had suffered stink-bug injury.

Analysis of the ethidium bromide bound to DNA by photoacoustic and FTIR spectroscopy

Under physiological conditions B-form DNA is an exceedingly stable structure. However, experimental evidences obtained through nuclear magnetic resonance and fluorescence anisotropy suggest that the structure of the double helix fluctuates substantially. We describe photoacoustic phase modulation frequency measurements of ethidium bromide (Eb) with calf thymus, DNA. As in fluorescence phase modulation measurements, we used an intercalating dye as a probe; however, we monitored the triplet excited state lifetime at different ionic strengths. The triplet lifetime of Eb varied from about 0.30 ms, with no DNA present, to 20 ms, (at a DNA:Eb molar ratio of 5). With salt titration, this value falls, to about 2.0 ms. This result suggests, a strong coupling between the phenantridinium ring of the ethidium and the base pairs because of the stacking movement of the DNA molecule under salt effect. This, effect may be understood considering DNA as a polyelectrolyte. The counterions, in the solution shield the phosphate groups, reducing the electrostatic repulsion force between them, hence compacting the DNA molecule. The results from Fourier transform infrared demonstrated two important bands: 3187 cm(-1) corresponding to the symmetric stretching of the NH group of the bases, and 1225 cm(-1) corresponding to the asymmetric stretching of phosphate groups shifted toward higher wavenumbers, suggesting a proximity between the intercalant and base pairs and a modification of the DNA backbone state, both induced by salt accretion.

A new biophysics approach using photoacoustic spectroscopy to study the DNA-ethidium bromide interaction

We have examined the binding processes of ethidium bromide interacting with calf thymus DNA using photoacoustic spectroscopy. These binding processes are generally investigated by a combination of absorption or fluorescence spectroscopies with hydrodynamic techniques. The employment of photoacoustic spectroscopy for the DNA-ethidium bromide system identified two binding manners for the dye. The presence of two isosbestic points (522 and 498 nm) during DNA titration was evidence of these binding modes. Analysis of the photoacoustic amplitude signal data was performed using the McGhee-von Hippel excluded site model. The binding constant obtained was 3.4 x 10(8) M(bp)(-1), and the number of base pairs excluded to another dye molecule by each bound dye molecule (n) was 2. A DNA drug dissociation process was applied using sodium dodecyl sulfate to elucidate the existence of a second and weaker binding mode. The dissociation constant determined was 0.43 mM, whose inverse value was less than the previously obtained binding constant, demonstrating the existence of the weaker binding mode. The calculated binding constant was adjusted by considering the dissociation constant and its new value was 1.2 x 10(9) M(bp)(-1) and the number of excluded sites was 2.6. Using the photoacoustic technique it is also possible to obtain results regarding the dependence of the quantum yield of the dye on its binding mode. While intercalated between two adjacent base pairs the quantum yield found was 0.87 and when associated with an external site it was 0.04. These results reinforce the presence of these two binding processes and show that photoacoustic spectroscopy is more extensive than commonly applied spectroscopies.

Calcium-Independent Membrane Damage by Venom Phospholipases A(2)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae).

NARANJO N, MONTERO DAV, SAENZ APONTE A. 2011. First record of infection by entomopathogenic nematodes of the grass bug Collaria scenica Stal (Hemiptera: Miridae). ENTOMOTROPICA 26(3): 117-125. The study was aimed to test the pathogenicity of Steinernema sp. and Heterorhabditis sp. in Collaria scenica. The effect of different concentrations of infective juveniles (IJ) were tested on nymphs and adults of C. scenica. For this purpose, the bugs were inoculated with 5 000 JI of each nematode species in a factorial design (3x2), and seven concentrations were tested in a JI factorial design (7x2x2). The bugs showed 100% mortality and symptoms of pathogenicity. Infection was found with both species of nematodes and penetration was assumed to be through the spiracles and anus. A higher capacity of pathogenicity was observed with Steinernema sp. Based on the results Heterorhabditis sp. and Steinernema sp. could constitute an efficient tool to control populations of C. scenica in pastures.

Gênese e Classificação da Coquina de Camaquã, Formação Corumbataí (Neopermiano), na Região de Rio Claro, SP

Analysis of the taphonomic signatures of a well preserved, silicifled coquina (Pinzonella neotropica assemblage) from the Camaquã outcrop, upper part of the Corumbataí Formation (Late Permian), in the Rio Claro region, state of Sáo Paulo, allowed interpretation of processes involved in its origin as related to high energy events (storms). The coquina occurs as a lenticular body, 2-11 cm thick and extending laterally for about 120 m. Basal contact of the coquina is sharp and erosive. Its upper contact is sharp. The concentration is dominated by pelecypods including the shallow burrowers (Pinzonella neotropica, Jacquesia brasiliensis), intermediate burrower (Pyramus anceps) and semi-infaunal forms (Naiadopsis lamellosus). All these species are suspension feeders. Besides sand-sized or even smaller shell fragments, there occur disarticulated, complete shells which are commonly abraded but do not show any signs of bioerosion or incrustation. In vertical side view, the shells are mainly convex-up, nested or stacked, while in plan-view they show random orientation. Multiple discontinuous grading is visible. These taphonomic signatures suggest that the origin of the skeletal accumulation is related to high energy events (possibly storm flows) in a proximal environment. The amalgamated nature of the Camaquã coquina records several episodes of erosion and deposition.

A new permian bivalve (Megadesmidae, Plesiocyprinellinae) from the Serrinha member, Rio do Rasto Formation, Paraná Basin, Brazil

During a recent inspection in the Paleontological Collection of the Institute of Geosciences, University of São Paulo, we have identifi ed some specimens of undescribed mollusk bivalves. These called our attention for the following reasons: a) all specimens are internal molds of conjugated and closed articulated valves, some of them presenting fragments of silicifi ed shells; b) all internal molds have similar general shape and internal characters, representing specimens of the same taxon; c) the internal molds and silicifi ed valves are well preserved, including fragile structures, which are hardly preserved, such as the internal mold of the external ligament and muscle scars; d) and equally important, according to the labels of all specimens, they were collected from rocks of the Passa Dois Group (Permian), Serrinha Member of the Rio do Rasto Formation. Although who collected the shells and the precise geographic location of the specimens are still unknown, the detailed study of these fossils brings us to the conclusion that they are morphologically distinct from any heretofore published genus of the endemic fauna of bivalves from Passa Dois Group. Based in its general shape, hinge structure and muscles scars, the new form can be classifi ed under the Family Megadesmidae Vokes, 1967, the most diverse group of Permian bivalves of the Paraná Basin. The specimens are referred as Beurlenella elongatella new gen. and sp. The shell shape and taphonomy indicate that this bivalve was a shallow, rapid, active burrower, suspension feeder, probably preserved in situ, in event deposits.

Herbivoria em soja: Efeito na composição química das folhas e na biologia da lagarta da soja e do percevejo verde pequeno

The objective of this research was to study the effect of previous herbivory in the chemical composition of soybean leaves, and its influence on Anticarsia gemmatalis (Hüeb.) and Piezodorus guildinii (West.) development. Two soybean genotypes, 'Embrapa 4' and 'BR 37', were chemically analyzed. The caterpillars were fed on these genotypes leaves and the bugs on string beans, both injured and non- injured (control), totalizing 4 treatments with 50 replications for each treatment. Herbivory was performed by releasing 3rd instar velvetbean caterpillar. The caterpillars were taken out when 30%-40% of foliar area were consumed. After that, leaves and string beans were used in the experiment. The results obtained during this research indicate that leaves chemical composition was altered and that, in general, A. gemmatalis larvae and P. guildinii nymph have presented better development when fed on non-injured soybean plants, being 'Embrapa 4' genotype the most efficient one concerning insects development.

Comparison of eggs, larvae, and pupae of plutella xylostella (lepidoptera: Plutellidae) as prey for podisus nigrispinus (Hemiptera: Pentatomidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Control de la chinche de los pastos Collaria scenica (Hemiptera: Miridae) con nematodos entomopatógenos en invernadero

We studied the efficacy of infective juveniles (IJ) of Steinernema sp. strain JCL024 and Heterorhabditis sp. strain SL0708 on adults and nymphs of Collaria scenica under greenhouse conditions. Five doses were evaluated (2, 16, 78, 160 IJ) cm-2 leaf) for each nematode. The experiment was evaluated for 21 days every 24 hours, making mortality counts and assessment of damage on Kikuyo grass. The bugs showed 100% mortality after 15 days. Each bug produced an average of 13,000 IJ/6 days. As for the damage did not reach level 2 within 21 days prior to treatment without nematodes that damage introduced 3 with necrosis and apical leaf curl. Thus, Steinernema sp. strain JCL024 and Heterorhabditis sp. strain SL0708 exercised control bug on the grass and kept under the level of damage to the foliage.