Combinatorial synthesis and directed evolution applied to the production of alpha-helix forming antimicrobial peptides analogues

Antimicrobial peptides (AMPs) are effector molecules of innate immune systems found in different groups of organisms, including microorganisms, plants, insects, amphibians and humans. These peptides exhibit several structural motifs but the most abundant AMPs assume an amphipathic alpha-helical structure. The alpha-helix forming antimicrobial peptides are excellent candidates for protein engineering leading to an optimization of their biological activity and target specificity. Nowadays several approaches are available and this review deals with the use of combinatorial synthesis and directed evolution in order to provide a high-throughput source of antimicrobial peptides analogues with enhanced lytic activity and specificity.

Accountability and revenue metering in smart micro-grids

Smart micro-grids offer a new challenging domain for power theories and metering techniques, because they include a variety of intermittent power sources which positively impact on power flow and distribution losses, but may cause voltage asymmetry and frequency variation. Due to the limited power capability of smart micro-grids, the voltage distortion can also get worse (in case of supplying non-linear loads), affecting measurement accuracy and possibly causing tripping of protections. In such a context, a reconsideration of power theories is required, since they form the basis for supply and load characterization. A revision of revenue metering techniques is also needed, to ensure a correct penalization of the loads for their responsibility in generating reactive power, voltage unbalance and distortion. This paper shows that the Conservative Power Theory (CPT) provides a suitable background to cope with smart grids characterization and metering needs. Experimental results validate the proposed approach. © 2010 IEEE.

Application of Conservative Power Theory to load and line characterization and revenue metering

This paper proposes a set of performance factors for load characterization and revenue metering. They are based on the Conservative Power Theory, and each of them relates to a specific load non-ideality (unbalance, reactivity, distortion). The performance factors are capable to characterize the load under different operating conditions, considering also the effect of non-negligible line impedances and supply voltage deterioration. © 2012 IEEE.

Dynamics and Conformational Studies of TOAC Spin Labeled Analogues of Ctx(Ile21)-Ha Peptide from Hypsiboas albopunctatus

Antimicrobial peptides (AMPs) isolated from several organisms have been receiving much attention due to some specific features that allow them to interact with, bind to, and disrupt cell membranes. The aim of this paper was to study the interactions between a membrane mimetic and the cationic AMP Ctx(Ile21)-Ha as well as analogues containing the paramagnetic amino acid 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) incorporated at residue positions n = 0, 2, and 13. Circular dichroism studies showed that the peptides, except for [TOAC13]Ctx(Ile21)-Ha, are unstructured in aqueous solution but acquire different amounts of α-helical secondary structure in the presence of trifluorethanol and lysophosphocholine micelles. Fluorescence experiments indicated that all peptides were able to interact with LPC micelles. In addition, Ctx(Ile21)-Ha and [TOAC13]Ctx(Ile21)-Ha peptides presented similar water accessibility for the Trp residue located near the N-terminal sequence. Electron spin resonance experiments showed two spectral components for [TOAC0]Ctx(Ile21)-Ha, which are most likely due to two membrane-bound peptide conformations. In contrast, TOAC2 and TOAC13 derivatives presented a single spectral component corresponding to a strong immobilization of the probe. Thus, our findings allowed the description of the peptide topology in the membrane mimetic, where the N-terminal region is in dynamic equilibrium between an ordered, membrane-bound conformation and a disordered, mobile conformation; position 2 is most likely situated in the lipid polar head group region, and residue 13 is fully inserted into the hydrophobic core of the membrane. © 2013 Vicente et al.

Dimerization of aurein 1.2: Effects in structure, antimicrobial activity and aggregation of Cândida albicans cells

Antimicrobial peptides (AMPs) are a promising solution to face the antibiotic-resistant problem because they display little or no resistance effects. Dimeric analogues of select AMPs have shown pharmacotechnical advantages, making these molecules promising candidates for the development of novel antibiotic agents. Here, we evaluate the effects of dimerization on the structure and biological activity of the AMP aurein 1.2 (AU). AU and the C- and N-terminal dimers, (AU)2K and E(AU)2, respectively, were synthesized by solid-phase peptide synthesis. Circular dichroism spectra indicated that E(AU)2 has a coiled coil structure in water while (AU)2K has an α-helix structure. In contrast, AU displayed typical spectra for disordered structures. In LPC micelles, all peptides acquired a high amount of α-helix structure. Hemolytic and vesicle permeabilization assays showed that AU has a concentration dependence activity, while this effect was less pronounced for dimeric versions, suggesting that dimerization may change the mechanism of action of AU. Notably, the antimicrobial activity against bacteria and yeast decreased with dimerization. However, dimeric peptides promoted the aggregation of C. albicans. The ability to aggregate yeast cells makes dimeric versions of AU attractive candidates to inhibit the adhesion of C. albicans to biological targets and medical devices, preventing disease caused by this fungus. © 2013 Springer-Verlag Wien.

Síntese, estudos estruturais e biológicos do peptídeo hilina a1 e análogos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Novel Antimicrobial Peptides Bacterial Cellulose Obtained by Symbioses Culture Between Polyhexanide Biguanide (PHMB) and Green Tea

Bacterial cellulose is a highly hydrated pellicle made up of a random assembly of ribbon shaped fibers less than 5 nm wide. The unique properties provided by the nanometric structure have led to a number of diagnostic biological probes, display devices due to their unique size-dependent medical applications. Bacterial cellulose matrix extracellular is a novel biotechnology and unique medicine indicated for ultimate chronic wound treatment management, drug delivery, tissue engineering, skin cancer and offers an actual and effective solution to a serious medical and social problem and to promote rapid healing in lesions caused by Diabetic burns, ulcers of the lower limbs or any other circumstance in which there's epidermal or dermal loss. In this work, it is reported novel antimicrobial peptides (AMPs) bacterial cellulose/polyhexanide biguanide (PHMB) which are produced by symbioses culture between polyhexanide biguanide and green tea culture medium resulting in the pure 3-D structure consisting of an ultra-fine network of novel biocellulose/PHMB nanofibres matrix (2-8 nm), highly hydrated (99% in weight), and with higher molecular weight, full biocompatibility.

Comprehensive Analysis of Gene Expression Profiles of the Beet Armyworm Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Vip3Aa Toxin

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Percepção de professores sobre a Avaliação de Habilidades Motoras e de Processo Versão Escolar aplicada aos alunos com deficiência física

Objetivou-se verificar sob a ótica do professor se a Avaliação de Habilidades Motoras e de Processo – Versão Escolar (School - Version of the Assessment of Motor and Process Skills - School-AMPS) aplicada aos alunos com deficiência física prediz o desempenho do aluno observado em sala de aula. Participaram dez crianças com deficiência física, entre quatro e oito anos de idade, e seus respectivos educadores. Os dados foram coletados por meio da observação das crianças em ambiente clínico durante a realização de tarefas pré-determinadas, com base na versão brasileira da School-AMPS. Aos educadores foi apresentado um relatório de avaliação da criança e aplicado um questionário composto por três questões. Os dados foram analisados de forma quantitativa e qualitativa. Os resultados apontaram que a School-AMPS possibilitou identificar se o desempenho da criança foi, na sua maioria, considerado deficitário, satisfatório ou intermediário e, sobretudo, possibilitou analisar o desempenho a partir da percepção da relação existente entre demandas da pessoa, da tarefa e do ambiente. Os educadores consideraram que o relatório de avaliação apresentado prediz de forma compatível o desempenho do aluno observado em sala de aula. Consideraram viáveis os recursos pedagógicos adaptados e as sugestões de estratégias para minimizar e/ou superar as dificuldades observadas no desempenho do aluno. Conclui-se que a versão brasileira da School-AMPS tem potencial como medida de desempenho funcional em crianças com deficiência física, entre quatro e oito anos de idade, e pode contribuir para a atuação do terapeuta ocupacional no contexto escolar.

Novos peptídeos sintéticos e estudo da interação com membranas modelo: efeito de modificações no N-terminal na atividade lítica e antimicrobiana

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos da dimerização na estrutura e atividade biológica dos peptídeos antimicrobianos Aureína 1.2 e Magainina 2

Pós-graduação em Biotecnologia - IQ

Structure-activity relationship of mastoparan analogs: effects of the number and positioning of Lys residues on secondary structure, interaction with membrane-mimetic systems and biological activity

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

Fatores microbiológicos e imunológicos envolvidos no desenvolvimento da cárie precoce da infância

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

Headgroup specificity for the interaction of the antimicrobial peptide tritrpticin with phospholipid Langmuir monolayers

We examined the interaction of the cationic antimicrobial peptide (AMP) tritrpticin (VRRFPWWWPFLRR, TRP3) with Langmuir monolayers of zwitterionic (dipalmitoyl phosphatidylcholine, DPPC, and dipalmitoyl phosphatidylethanolamine, DPPE) and negatively charged phospholipids (dipalmitoyl phosphatidic acid, DPPA, and dipalmitoyl phosphatidylglycerol, DPPG). Both surface pressure and surface potential isotherms became more expanded upon addition of TRP3 (DPPE similar to DPPC << DPPA < DPPG). The stronger interaction with negatively charged phospholipids agrees with data for vesicles and planar lipid bilayers, and with AMPs greater activity against bacterial membranes versus mammalian cell membranes. Considerable expansion of negatively charged monolayers occurred at 10 and 30 mol% TRP3, especially at low surface pressure. Moreover, a difference was observed between PA and PG, demonstrating that the interaction, besides being modulated by electrostatic interactions, displays specificity with regard to headgroup, being more pronounced in the case of PG, present in large quantities in bacterial membranes. In previous studies, it was proposed that the peptide acts by a toroidal pore-like mechanism [1,2]. Considering the evidence from the literature that PG shows a propensity to form a positive curvature as do toroidal pores, the observation of TRP3's preference for the PG headgroup and the dramatic increase in area promoted by this interaction represent further support for the toroidal pore mechanism of action proposed for TRP3. (C) 2012 Elsevier B.V. All rights reserved.

Optical transition radiation used in the diagnostic of low energy and low current electron beams in particle accelerators

Optical transition radiation (OTR) plays an important role in beam diagnostics for high energy particle accelerators. Its linear intensity with beam current is a great advantage as compared to fluorescent screens, which are subject to saturation. Moreover, the measurement of the angular distribution of the emitted radiation enables the determination of many beam parameters in a single observation point. However, few works deals with the application of OTR to monitor low energy beams. In this work we describe the design of an OTR based beam monitor used to measure the transverse beam charge distribution of the 1.9-MeV electron beam of the linac injector of the IFUSP microtron using a standard vision machine camera. The average beam current in pulsed operation mode is of the order of tens of nano-Amps. Low energy and low beam current make OTR observation difficult. To improve sensitivity, the beam incidence angle on the target was chosen to maximize the photon flux in the camera field-of-view. Measurements that assess OTR observation (linearity with beam current, polarization, and spectrum shape) are presented, as well as a typical 1.9-MeV electron beam charge distribution obtained from OTR. Some aspects of emittance measurement using this device are also discussed. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.4748519]