Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

A novel device with 36 channels for imaging and signal acquisition of the gastrointestinal tract based on AC biosusceptometry

The alternate current biosusceptometry (ACB) is a biomagnetic technique used to study some physiological parameters associated with gastrointestinal (GI) tract. For this purpose it applies an AC magnetic field and measures the response originating from magnetic marks or tracers. This paper presents an equipment based on the ACB which uses anisotropic magnetoresistive (AMR) sensors and an inexpensive electronic support. The ACB-AMR developed consists of a square array of 6x6 sensors arranged in a firstorder gradiometer configuration with one reference sensor. The equipment was applied to capture magnetic images of different phantoms and to acquire gastric contraction activity of healthy rats. The results show a reasonable sensitivity and spatial-temporal resolution, so that it may be applied for imaging of phantoms and signal acquisition of the GI tract of small animals. © 2010 IEEE.

Chemical and biological characterization of four new linear cationic α-helical peptides from the venoms of two solitary eumenine wasps

Four novel peptides were isolated from the venoms of the solitary eumenine wasps Eumenes rubrofemoratus and Eumenes fraterculus. Their sequences were determined by MALDI-TOF/TOF (matrix assisted laser desorption/ionization time-of-flight mass spectrometry) analysis, Edman degradation and solid-phase synthesis. Two of them, eumenitin-R (LNLKGLIKKVASLLN) and eumenitin-F (LNLKGLFKKVASLLT), are highly homologous to eumenitin, an antimicrobial peptide from a solitary eumenine wasp, whereas the other two, EMP-ER (FDIMGLIKKVAGAL-NH 2) and EMP-EF (FDVMGIIKKIAGAL-NH 2), are similar to eumenine mastoparan-AF (EMP-AF), a mast cell degranulating peptide from a solitary eumenine wasp. These sequences have the characteristic features of linear cationic cytolytic peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, they can be predicted to adopt an amphipathic α-helix secondary structure. In fact, the CD (circular dichroism) spectra of these peptides showed significant α-helical conformation content in the presence of TFE (trifluoroethanol), SDS (sodium dodecylsulfate) and asolectin vesicles. In the biological evaluation, all the peptides exhibited a significant broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. © 2011 Elsevier Ltd.

Chemometric analysis of Hymenoptera toxins and defensins: A model for predicting the biological activity of novel peptides from venoms and hemolymph

When searching for prospective novel peptides, it is difficult to determine the biological activity of a peptide based only on its sequence. The trial and error approach is generally laborious, expensive and time consuming due to the large number of different experimental setups required to cover a reasonable number of biological assays. To simulate a virtual model for Hymenoptera insects, 166 peptides were selected from the venoms and hemolymphs of wasps, bees and ants and applied to a mathematical model of multivariate analysis, with nine different chemometric components: GRAVY, aliphaticity index, number of disulfide bonds, total residues, net charge, pI value, Boman index, percentage of alpha helix, and flexibility prediction. Principal component analysis (PCA) with non-linear iterative projections by alternating least-squares (NIPALS) algorithm was performed, without including any information about the biological activity of the peptides. This analysis permitted the grouping of peptides in a way that strongly correlated to the biological function of the peptides. Six different groupings were observed, which seemed to correspond to the following groups: chemotactic peptides, mastoparans, tachykinins, kinins, antibiotic peptides, and a group of long peptides with one or two disulfide bonds and with biological activities that are not yet clearly defined. The partial overlap between the mastoparans group and the chemotactic peptides, tachykinins, kinins and antibiotic peptides in the PCA score plot may be used to explain the frequent reports in the literature about the multifunctionality of some of these peptides. The mathematical model used in the present investigation can be used to predict the biological activities of novel peptides in this system, and it may also be easily applied to other biological systems. © 2011 Elsevier Inc.

Wavelets in a problem of signal processing

In this work a new method is proposed for noise reduction in speech signals in the wavelet domain. The method for signal processing makes use of a transfer function, obtained as a polynomial combination of three processings, denominated operators. The proposed method has the objective of overcoming the deficiencies of the thresholding methods and the effective processing of speech corrupted by real noises. Using the method, two speech signals are processed, contaminated by white noise and colored noises. To verify the quality of the processed signals, two evaluation measures are used: signal to noise ratio (SNR) and perceptual evaluation of speech quality (PESQ).

Agelaia MP-I: A peptide isolated from the venom of the social wasp, Agelaia pallipes pallipes, enhances insulin secretion in mice pancreatic islets

Peptides isolated from animal venoms have shown the ability to regulate pancreatic beta cell function. Characterization of wasp venoms is important, since some components of these venoms present large molecular variability, and potential interactions with different signal transduction pathways. For example, the well studied mastoparan peptides interact with a diversity of cell types and cellular components and stimulate insulin secretion via the inhibition of ATP dependent K + (K ATP) channels, increasing intracellular Ca 2+ concentration. In this study, the insulin secretion of isolated pancreatic islets from adult Swiss mice was evaluated in the presence of synthetic Agelaia MP-I (AMP-I) peptide, and some mechanisms of action of this peptide on endocrine pancreatic function were characterized. AMP-I was manually synthesized using the Fmoc strategy, purified by RP-HPLC and analyzed using ESI-IT-TOF mass spectrometry. Isolated islets were incubated at increasing glucose concentrations (2.8, 11.1 and 22.2 mM) without (Control group: CTL) or with 10 μM AMP-I (AMP-I group). AMP-I increased insulin release at all tested glucose concentrations, when compared with CTL (P < 0.05). Since molecular analysis showed a potential role of the peptide interaction with ionic channels, insulin secretion was also analyzed in the presence of 250 μM diazoxide, a K ATP channel opener and 10 μM nifedipine, a Ca 2+ channel blocker. These drugs abolished insulin secretion in the CTL group in the presence of 2.8 and 11.1 mM glucose, whereas AMP-I also enhanced insulin secretory capacity, under these glucose conditions, when incubated with diazoxide and nifedipine. In conclusion, AMP-I increased beta cell secretion without interfering in K ATP and L-type Ca 2+ channel function, suggesting a different mechanism for this peptide, possibly by G protein interaction, due to the structural similarity of this peptide with Mastoparan-X, as obtained by modeling. © 2012 Elsevier Ltd.

Components of the acoustic swallowing signal: Preliminary study

Purpose: To analyze the components of the acoustic signal of swallowing using a specific software. Methods: Fourteen healthy subjects ranging in age from 20 to 50 years (mean age 31±10 years), were evaluated. Data collection consisted on the simultaneous capture of the swallowing audio with a microphone and of the swallowing videofluoroscopic image. The bursts of the swallowing acoustic signal were identified and their duration and the interval between them were later analyzed using a specific software, which allowed the simultaneous analyses between the acoustic wave and the videofluoroscopic image. Results: Three burst components were identified in most of the swallows evaluated. The first burst presented mean time of 87.3 milliseconds (ms) for water and 78.2 for the substance. The second burst presented mean time of 112.9 ms for water and 85.5 for the pasty substance. The mean interval between first and second burst was 82.1 ms for water and 95.3 ms for the pasty consistency, and between second and third burst was 339.8 ms for water and 322.0 ms for the pasty consistency. Conclusion: The software allowed the visualization of three bursts during the swallowing of healthy individuals, and showed that the swallowing signal in normal subjects is highly variable.

Canonical WNT signaling regulates development of bovine embryos to the blastocyst stage

Objectives were to evaluate the role of canonical WNT signaling in development of the preimplantation embryo. Signaling was activated with 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP) and inhibited with Dickkopf-related protein 1 (DKK1). Treatment of bovine embryos with AMBMP at day 5 after insemination decreased development to the blastocyst stage at day 7 and reduced numbers of trophectoderm and inner cell mass cells. At high concentrations, AMBMP caused disorganization of the inner cell mass. DKK1 blocked actions of AMBMP but did not affect development in the absence of AMBMP. Examination of gene expression in day 6 morulae by microarray revealed expression of 16 WNT genes and other genes involved in WNT signaling; differences in relative expression were confirmed by PCR for 7 genes. In conclusion, the preimplantation embryo possesses a functional WNT signaling system and activation of the canonical pathway can inhibit embryonic development.

Signal classification of submerged aquatic vegetation based on the hemispherical-conical reflectance factor spectrum shape in the yellow and red regions

The water column overlying the submerged aquatic vegetation (SAV) canopy presents difficulties when using remote sensing images for mapping such vegetation. Inherent and apparent water optical properties and its optically active components, which are commonly present in natural waters, in addition to the water column height over the canopy, and plant characteristics are some of the factors that affect the signal from SAV mainly due to its strong energy absorption in the near-infrared. By considering these interferences, a hypothesis was developed that the vegetation signal is better conserved and less absorbed by the water column in certain intervals of the visible region of the spectrum; as a consequence, it is possible to distinguish the SAV signal. To distinguish the signal from SAV, two types of classification approaches were selected. Both of these methods consider the hemispherical-conical reflectance factor (HCRF) spectrum shape, although one type was supervised and the other one was not. The first method adopts cluster analysis and uses the parameters of the band (absorption, asymmetry, height and width) obtained by continuum removal as the input of the classification. The spectral angle mapper (SAM) was adopted as the supervised classification approach. Both approaches tested different wavelength intervals in the visible and near-infrared spectra. It was demonstrated that the 585 to 685-nm interval, corresponding to the green, yellow and red wavelength bands, offered the best results in both classification approaches. However, SAM classification showed better results relative to cluster analysis and correctly separated all spectral curves with or without SAV. Based on this research, it can be concluded that it is possible to discriminate areas with and without SAV using remote sensing. © 2013 by the authors; licensee MDPI, Basel, Switzerland.

Periapical lesions decrease insulin signal and cause insulin resistance

Introduction: Inflammatory cytokines are associated with decreased insulin signal transduction. Moreover, local oral inflammation, such as that accompanying periodontal disease, is associated with insulin resistance and type 2 diabetes mellitus. The aim of this study was to evaluate the effect of periapical lesions (PLs) on insulin signaling and insulin sensitivity in rats. We hypothesized that PLs alter systemic insulin signaling and insulin sensitivity via elevated plasmatic tumor necrosis factor α (TNF-α). Methods: Wistar rats were divided into control (CN) and PL groups. PLs were induced by exposing pulpal tissue to the oral environment. After 30 days, insulin sensitivity was measured using the insulin tolerance test. After euthanization, maxillae were processed for histopathology. Plasmatic concentrations of tumor necrosis factor α (TNF-α) were determined via the enzyme-linked immunosorbent assay. Insulin signal transduction was evaluated using insulin receptor substrate tyrosine phosphorylation status and serine phosphorylation status in periepididymal white adipose tissue via Western blotting. For insulin signaling and insulin tolerance tests, the analyses performed were analysis of variance followed by the Tukey post hoc test. For TNF-α analysis, the Student's t test was used. In all tests, P <.05 was considered significant. Results: The rats with PLs showed higher plasmatic TNF-α, lower constant rate for glucose disappearance values, and reduced pp185 tyrosine phosphorylation status but no change in serine phosphorylation status in white adipose tissue after insulin stimulation. Conclusions: PLs can cause alterations to both insulin signaling and insulin sensitivity, probably because of elevation of plasmatic TNF-α. The results from this study emphasize the importance of the prevention of local inflammatory diseases, such as PLs, with regard to the prevention of insulin resistance. Copyright © 2013 American Association of Endodontists.

Yield and mineral nutrition of soybean, maize, and congo signal grass as affected by limestone and slag

The objective of this work was to evaluate the efficiency of superficial applicationof limestone and slag, and their effects on soil chemical attributes and on yield and mineral nutrition of soybean, maize, and Congo signal grass (Urochloa ruziziensis). The experiment was carried out in a Rhodic Hapludox under no tillage system. The treatments consisted of the use of limestone or slag (silicates of calcium and magnesium) to correct soilacidity, and of a control treatment without the use of soil correctives. Rates were calculated in order to raise soil base saturation up to 70%. Soybean was sown in November 2006and maize in December 2007. Congo signal grass was sown right after the harvests of soybean and maize, and it was cropped during the offseasons. Soil chemical attributes were evaluated at 6, 12, and 18 months after the application of the corrective materials. Slag isan efficient source for soil acidity correction, being able to raise the exchangeable base levels in the soil profile faster than lime. Both limestone and slag increase dry matter yield of Congo signal grass, and grain yield of soybean and maize. Slag is more effective in improving maize grain yield.

How a simple and stereotyped acoustic signal transmits individual information: the song of the White-browed Warbler Basileuterus leucoblepharus

O Pula-pula-assobiador Basileuterus leucoblepharus, um pássaro comum da Mata Atlântica, emite um único e distintivo tipo de canto para defesa territorial. O reconhecimento individual ou entre vizinho e estranho pode ser mais difícil quando as aves compartilham cantos semelhantes. De fato, a análise dos cantos de diferentes indivíduos revelou ligeiras diferenças nos domínios temporal e das freqüências. Efetivamente, um exame cuidadoso dos sinais de 21 indivíduos diferentes por 5 métodos complementares de análise revelou que, primeiro, um ou dois espaços na série tonal ocorrem entre duas notas sucessivas em determinados momentos do canto e, segundo, ocupam posições em tempo e freqüência estereotipadas para cada indivíduo. Experiências de "play-back" confirmam esses dados. Através de experiências de propagação, mostramos que esta informação individual pode ser transmitida somente a curta distância ( < 100 m) na mata. Considerando o tamanho e a repartição dos territórios, este processo de comunicação mostra-se eficiente e bem adaptado.

Defoliation patterns in signal grass tillers with varying heights in the same pasture

The experiment was carried out to evaluate the patterns of defoliation in plants of various heights in the same pasture of Brachiaria decumbens cv. Basilisk under continuous stocking with cattle. Four plant heights were evaluated (10, 20, 30 and 40 cm) in the same managed sward, with mean height equal to 25 cm. A randomized blocks design was used, with two replications. Defoliation frequency increased linearly along with plant height in the same pasture. Defoliation intensity, number of defoliations in leaf blade and percentage of leaf blade grazed per tiller also increased linearly along with plant height. The defoliation interval decreased linearly according to plant height. The quadratic model was adequately fitted into grazing efficiency data, reaching the highest value (98%) at the sward site with 35 cm plants. The percentage variation in signal grass height showed a negative linear response with initial plant height. The structure of the pasture, characterized by the diversity in plant heights, is simultaneously the cause and consequence of the variability in defoliation patterns in individual tillers, which occur on the horizontal plane of the pasture.

Initial height of pasture deferred and utilized in winter and tillering dynamics of signal grass during the following spring

Tillering dynamics during regrowth in the spring (September to December 2009) on signal grass (Brachiaria decumbens cv. Basilisk) pastures deferred in the beginning of April 2009 is evaluated. Four pasture heights were evaluated at the beginning of deferment (10, 20, 30 and 40 cm). The experimental design consisted of completely randomized blocks with two replications. Rise in pasture height at the beginning of deferment decreased the appearance rate, increased the mortality rate and reduced the survival rate, the balance between appearance and mortality and the stability index of the tiller population at the beginning of the spring. Pastures deferred at initial height of 30 and 40 cm presented relatively stable tiller appearance rate between the beginning and end of spring. However, these pastures presented greater mortality rate, lower survival rate, lower balance between appearance and mortality and lower stability index of the tiller population at the beginning of spring when compared to rates at the end of spring. Pastures managed with shorter initial heights (10 and 20 cm) provided more tissue renewal in the subsequent growth season after their utilization in the winter.