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.

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.

Thermography and ultrasonography in back pain diagnosis of equine athletes

The object of the current study was to evaluate the efficacy of thermography and ultrasonography in the diagnosis of thoracolumbar lesions in Quarter Horse athletes and associate the different types of lesions found with the athletic modality practiced. Twenty-four horses were admitted to the Surgery Service for Large Animals of the Veterinary and Animal Science Faculty, UNESP, Botucatu, Brazil, with complaints of back problems. All the horses were submitted for physical examinations to confirm the existence of thoracolumbar alterations and then for thermography and ultrasonography Thermography was used to map the lesioned areas of this region and ultrasonography for lesion characterization. The lesions found were supraspinous desmitis, interspinous desmitis, dorsal intervertebral osteoarthritis, and impingement of the spinous processes or kissing spines. The existence of a relation between the type of event practiced by the horse and the type of lesion found was determined. In horses that competed in the barrel race, a predominance of lesions in the thoracic caudal, thoracolumbar, and cranial lumbar regions occurred, with intervertebral osteoarthritis and interspinous desmitis being the most common. In cutting horses, most of the lesions were observed in the caudal lumbar region, whereas horses competing in reining showed a preferential location for lesions in the middle lumbar, with a predominance of supraspinous desmitis and myositis. Thermography associated with ultrasonography was shown to be efficient in the diagnosis of the thoracolumbar lesions of these horses.

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.

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.

Spatial Distribution of Euschistus heros (F.) (Hemiptera: Pentatomidae) in Soybean

Soybean bugs are major crop pests that cause significant reduction in harvest yield and influence grain quality. The aim of this study was to verify the spatial distribution of Euschistus heros (F.) (Hemiptera: Pentatomidae) in conventional and transgenic soybean cultivars. The experiment was conducted during the 2010-2011 crop season in UNESP/FCAV, Jaboticabal, SP, Brazil, in two fields of 10,000-m2 area that were subdivided into 100 plots (10 m × 10 m). The cultivars sown were M 7908 RR and its isoline M-SOY 8001. The number of the first to fifth instars and the number of adults were determined. To evaluate insect dispersion in the area, the following indices were used: variance/mean ratio, Morisita index, Green coefficient, and the k exponent of the negative binomial distribution. To study probabilistic models to describe the spatial distribution of the insects, the adjustments of the Poisson and negative binomial distributions were tested. The first to third instars showed aggregated spatial distribution, whereas the fourth and fifth instars, and adults, isolated or grouped, showed variation in the arrangement, ranging from moderately aggregated to randomly dispersed. During the adjustment of probability distributions, the negative binomial distribution model showed adjustment for the first to third instars, fourth and fifth instars, adults, and fourth and fifth instars plus adults. © 2013 Sociedade Entomológica do Brasil.

Interações de aranhas (Araneae) e artrópodes-praga (Acari e Hemiptera) em cultivos comerciais de seringueira (Hevea brasiliensis) no Noroeste do Estado de São Paulo

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

Avaliação do potencial tripanocida de substâncias extraídas da Família Piperaceae em formas epimastigotas de Trypanosoma cruzi

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Termografia e ultra-sonografia no diagnóstico de lesões toracolombares em eqüinos atletas da raça Quarto de Milha

Pós-graduação em Medicina Veterinária - FMVZ