Study of the mechanism of action of anoplin, a helical antimicrobial decapeptide with ion channel-like activity, and the role of the amidated C-terminus

Anoplin, an antimicrobial, helical decapeptide from wasp venom, looses its biological activities by mere deamidation of its C-terminus. Secondary structure determination, by circular dichroism spectroscopy in amphipathic environments, and lytic activity in zwitterionic and anionic vesicles showed quite similar results for the amidated and the carboxylated forms of the peptide. The deamidation of the C-terminus introduced a negative charge at an all-positive charged peptide, causing a loss of amphipathicity, as indicated by molecular dynamics simulations in TFE/water mixtures and this subtle modification in a peptide's primary structure disturbed the interaction with bilayers and biological membranes. Although being poorly lytic, the amidated form, but not the carboxylated, presented ion channel-like activity on anionic bilayers with a well-defined conductance step; at approximately the same concentration it showed antimicrobial activity. The pores remain open at trans-negative potentials, preferentially conducting cations, and this situation is equivalent to the interaction of the peptide with bacterial membranes that also maintain a high negative potential inside. Copyright (C) 2007 European Peptide Society and John Wiley & Sons, Ltd.

Study of the effect of the peptide loading and solvent system in SPPS by HRMAS-NMR

The SPPS methodology has continuously been investigated as a valuable model to monitor the solvation properties of polymeric materials. In this connection, the present work applied HRMAS-NMR spectroscopy to examine the dynamics of an aggregating peptide sequence attached to a resin core with varying peptide loading (up to 80%) and solvent system. Low and high substituted BHAR were used for assembling the VQAAIDYING sequence and some of its minor fragments. The HRMAS-NMR results were in agreement with the swelling of each resin, i.e. there was an improved resolution of resonance peaks in the better solvated conditions. Moreover, the peptide loading and the attached peptide sequence also affected the spectra. Strong peptide chain aggregation was observed mainly in highly peptide loaded resins when solvated in CDCl3. Conversely, due to the better swelling of these highly loaded resins in DMSO, improved NMR spectra were acquired in this polar aprotic solvent, thus enabling the detection of relevant sequence-dependent conformational alterations. The more prominent aggregation was displayed by the VQAAIDYING segment and not by any of its intermediary fragments and these findings were also corroborated by EPR studies of these peptide-resins labelled properly with an amino acid-type spin probe. Copyright (c) 2005 European Peptide Society and John Wiley & Sons, Ltd.

Effect of thermal cycling and disinfection on colour stability of denture base acrylic resin

Objectives: The purpose of this study was to investigate the effect of thermal cycling and disinfection on the colour change of denture base acrylic resin. Materials and Methods: Four different brands of acrylic resins were evaluated (Onda Cryl, QC 20, Classico and Lucitone). All brands were divided into four groups (n=7) determined according to the disinfection procedure (microwave, Efferdent, 4% chlorhexidine or 1% hypochlorite). The treatments were conducted three times a week for 60days. All specimens were thermal cycled between 5 and 55°C with 30-s dwell times for 1000 cycles before and after disinfection. The specimens' colour was measured with a spectrophotometer using the CIE L*a*b* system. The evaluations were conducted at baseline (B), after first thermal cycling (T 1), after disinfection (D) and after second thermal cycling (T 2). Colour differences (ΔE) were calculated between T 1 and B (T 1B), D and B (DB), and T 2 and B (T 2B) time-points. Results: The samples submitted to disinfection by microwave and Efferdent exhibited the highest values of colour change. There were significant differences on colour change between the time-points, except for the Lucitone acrylic resin. Conclusions: The thermal cycling and disinfection procedures significantly affected the colour stability of the samples. However, all values obtained for the acrylic resins are within acceptable clinical parameters. © 2012 The Gerodontology Society and John Wiley & Sons A/S.

Complete denture wearing and fractures among edentulous patients treated in university clinics

Objective: The prevalence of wearing and fracture of complete dentures was evaluated among edentulous patients treated in two dental schools in Brazil. Background: Acceptance and wearing of complete dentures are related to adaptive behaviour of edentulous patients. However, one reason that could interfere with the wearing dentures is their potential to fracture, which is still a common complication in denture rehabilitation practice. Material and methods: Two hundred and twenty-four edentulous patients rehabilitated with complete dentures from 2000 to 2005 in Araçatuba and Araraquara Dental School, University of State of São Paulo, were assessed in 2006 and 2007 to answer a questionnaire about wearing and fracture of their dentures. Statistical analysis were performed using Epi Info software and chi-squared test to compare maxillary and mandibular data (α = 0.05). Results: Almost 26% of the patients did not wear their dentures, and among the remainder, the majority wore the maxillary denture. About 30% of the dentures were fractured, with higher prevalence in the maxillary arch (p = 0.003). Conclusions: Discontinuation of wearing dentures was quite high, especially considering the treatment which was carried out in university clinics. Prevalence of fractures was also high, greater for the maxillary denture, and was one of the main reasons for non-wearing of complete dentures. © 2011 The Gerodontology Society and John Wiley & Sons A/S.

Antimicrobial activity and surface properties of an acrylic resin containing a biocide polymer

Objective: To evaluate the antimicrobial activity and surface properties of an acrylic resin containing the biocide polymer poly (2-tert-butylaminoethyl) methacrylate (PTBAEMA). Background: Several approaches have been proposed to prevent oral infections, including the incorporation of antimicrobial agents to acrylic resins. Materials and methods: Specimens of an acrylic resin (Lucitone 550) were divided into two groups: 0% (control) and 10% PTBAEMA. Antimicrobial activity was assessed by adherence assay of one of the microorganisms, Staphylococcus aureus, Streptococcus mutans and Candida albicans. Surface topography was characterised by atomic force microscopy and wettability properties determined by contact angle measurements. Results: Data of viable cells (log (CFU + 1)/ml) for S. aureus (control: 7.9 ± 0.8; 10%: 3.8 ± 3.3) and S. mutans (control: 7.5 ± 0.7; 10%: 5.1 ± 2.7) showed a significant decrease with 10% of PTBAEMA (Mann-Whitney, p < 0.05). For C. albicans (control: 6.6 ± 0.2; 10%: 6.6 ± 0.4), there was no significant difference between control and 10% of PTBAEMA (Kruskal-Wallis, p > 0.05). Incorporating 10% PTBAEMA increased surface roughness and decreased contact angles. Conclusion: Incorporating 10% PTBAEMA into acrylic resins increases wettability and roughness of acrylic resin surface; and decreases the adhesion of S. mutans and S. aureus on acrylic surface, but did not exhibit antimicrobial effect against C. albicans. © 2012 The Gerodontology Society and John Wiley & Sons A/S.

Effect of different disinfectants on the microhardness and roughness of acrylic resins for ocular prosthesis

Background: Ocular prosthesis materials should have specific properties for their indication and durability; therefore, it is important to investigate their physical behaviour when affected by several disinfectants. Objectives: This study evaluated the influence of different disinfecting solutions on the microhardness and surface roughness of acrylic resins for ocular prosthesis. Materials and Methods: Fifty samples simulating ocular prostheses were fabricated with N1 resin and colourless resin and divided (n = 10) according to the disinfectant used: neutral soap, Opti-free, Efferdent, 1% hypochlorite (HYC) and 4% chlorhexidine (CHX). Samples were stored in saline solution at 37°C and disinfected during 120 days. Both microhardness and roughness were investigated before, after 60 days and 120 days of disinfection and storage. Microhardness was measured using a microhardner and the roughness with a roughness device. Results: N1 resin showed lower microhardness when compared with colourless resin (p < 0.05). HYC and CHX groups exhibited the highest change of microhardness and roughness values (p < 0.05). An increase in roughness and reduction in microhardness of ocular acrylic resins were observed after both periods of disinfection and storage (p < 0.05). Conclusion: Both disinfection/storage periods affected the microhardness and roughness values of the samples. © 2012 The Gerodontology Society and John Wiley & Sons A/S.

Involvement of the insular cortex in the consolidation and expression of contextual fear conditioning

The insular cortex (IC) has been reported to be involved in the modulation of memory and autonomic and defensive responses. However, there is conflicting evidence about the role of the IC in fear conditioning. To explore the IC involvement in both behavioral and autonomic responses induced by contextual fear conditioning, we evaluated the effects of the reversible inhibition of the IC neurotransmission through bilateral microinjections of the non-selective synapse blocker CoCl2 (1 mm) 10 min before or immediately after the conditioning session or 10 min before re-exposure to the aversive context. In the conditioning session, rats were exposed to a footshock chamber (context) and footshocks were used as the unconditioned stimulus. Forty-eight hours later, the animals were re-exposed to the aversive context for 10 min, but no shock was given. Behavioral (freezing) as well as cardiovascular (arterial pressure and heart rate increases) responses induced by re-exposure to the aversive context were analysed. It was observed that the local IC neurotransmission inhibition attenuated freezing and the mean arterial pressure and heart rate increase of the groups that received the CoCl2 either immediately after conditioning or 10 min before re-exposure to the aversive context, but not when the CoCl2 was injected before the conditioning session. These findings suggest the involvement of the IC in the consolidation and expression of contextual aversive memory. However, the IC does not seem to be essential for the acquisition of memory associated with aversive context. © 2013 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

O neocontratualismo de Norberto Bobbio e John Rawls em cum contexto de neoliberalismo e crise estrutural do capital

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estrutura e significado em Uma rosa para Emily, de William Faulkner

Trata-se de uma análise do consagrado conto Uma Rosa para Emily, de William Faulkner, voltada para alguns dos principais aspectos de sua estrutura. Após considerarmos o enredo, discutimos a construção das personagens, com destaque para a protagonista, fazendo um levantamento e comentários sobre possíveis fontes de inspiração, destacando, entre outras, aspectos da biografia da poeta Emily Dickinson, a ficção e a poesia de E. A. Poe, romances de Charles Dickens e Henry James, o conto de Sherwood Anderson e a poesia de William Blake, Emily Dickinson, Robert Browning e John Crowe Ransom, acrescentando paralelos com o conto Bartleby, o escrivão, de Herman Melville. Analisamos, então, o foco narrativo, os símbolos e o significado, ressaltando aqui o desenvolvimento temático da narrativa.

Aprender versus ensinar: Charles Sanders Peirce e a universidade americana do final do século XIX

A produção científica e filosófica de Charles Sanders PEIRCE (1839-1914), exigindo como critério para o trabalho intelectual e para a conduta da vida do pensador o absoluto rigor na construção dos conceitos e a estrita verificação experimental, teve por conseqüência desvincular o trabalho científico e filosófico de qualquer função apologética. A afirmação de que todo conhecimento do mundo da experiência e mesmo daquele elaborado pela matemática é intrinsecamente provável e falível se opôs a todo e qualquer dogmatismo e mesmo ao a priori de tradição Kantiana. O interesse pela teoria evolucionista e a coerência inabalável da filosofia e das atitudes de PEIRCE, como professor e pesquisador, encontraram profunda resistência no meio universitário e editorial de seu tempo. Num momento de grave crise na Universidade norte-americana, decorrente das transformações econômicas e políticas ocorridas com a guerra da Secessão (1861-1865), o posicionamento de PEIRCE contribuiu muito provavelmente para sua demissão como professor das Universidades de Harvard e de John Hopkins; para dificultar a publicação de seus escritos e para seu total isolamento nos últimos anos de vida.

A 4.2 kDa synthetic peptide as a potential probe to evaluate the antibacterial activity of coumarin drugs

The coumarin antibiotics are potent inhibitors of DNA replication whose target is the enzyme DNA gyrase, an ATP-dependent bacterial type II topoisomerase. The coumarin drugs inhibit gyrase action by competitive binding to the ATP-binding site of DNA gyrase B protein. The production of new biologically active products has stimulated additional studies on coumarin-gyrase interactions. In this regard, a 4.2 kDa peptide mimic of DNA gyrase B protein from Escherichia coli has been designed and synthesized. The peptide sequence includes the natural fragment 131-146 (coumarin resistance-determining region) and a segment containing the gyrase-DNA interaction region (positions 753-770). The peptide mimic binds to novobiocin (K-a = 1.4 +/- 0.3 x 10(5) m(-1)), plasmid (K-a = 1.6 +/- 0.5 x 10(6) m(-1)) and ATP (K-a = 1.9 f 0.4 x 10(3) m(-1)), results previously found with the intact B protein. on the other hand, the binding to novobiocin was reduced when a mutation of Arg-136 to Leu-136 was introduced, a change previously found in the DNA gyrase B protein from several coumarin-resistant clinical isolates of Escherichia coLi. In contrast, the binding to plasmid and to ATP was not altered. These results suggest that synthetic peptides designed in a similar way to that described here could be used as mimics of DNA gyrase in studies which seek a better understanding of the ATP, as well as coumarin, binding to the gyrase and also the mechanism of action of this class of antibacterial drugs. Copyright (C) 2004 European Peptide Society and John Wiley Sons, Ltd.

Two short peptides including segments of subunit A of Escherichia coli DNA gyrase as potential probes to evaluate the antibacterial activity of quinolones

Quinolones constitute a family of compounds with a potent antibiotic activity. The enzyme DNA gyrase, responsible for the replication and transcription processes in DNA of bacteria, is involved in the mechanism of action of these drugs. In this sense, it is believed that quinolones stabilize the so-called 'cleavable complex' formed by DNA and gyrase, but the whole process is still far from being understood at the molecular level. This information is crucial in order to design new biological active products. As an approach to the problem, we have designed and synthesized low molecular weight peptide mimics of DNA gyrase. These peptides correspond to sequences of the subunit A of the enzyme from Escherichia coli, that include the quinolone resistance-determining region (positions 75-92) and a segment containing the catalytic Tyr-122 (positions 116-130). The peptide mimic of the non-mutated enzyme binds to ciprofloxin (CFX) only when DNA and Mg2+ were present (Kd = 1.6 × 10 -6 m), a result previously found with DNA gyrase. On the other hand, binding was reduced when mutations of Ser-83 to Leu-83 and Asp-87 to Asn-87 were introduced, a double change previously found in the subunit A of DNA gyrase from several CFX-resistant clinical isolates of E. coli. These results suggest that synthetic peptides designed in a similar way to that described here can be used as mimics of gyrases (topoisomerases) in order to study the binding of the quinolone to the enzyme-DNA complex as well as the mechanism of action of these antibiotics. Copyright © 2001 European Peptide Society and John Wiley & Sons, Ltd.

Candida albicans adherence to an acrylic resin modified by experimental photopolymerised coatings: An in vitro study

Purpose: This study evaluated whether photopolymerised coatings containing zwitterion or hydrophilic monomers would reduce the adhesion of Candida albicans to an acrylic resin. Materials and methods: Disc-shaped samples (n = 468) were fabricated with rough or smooth surfaces. The samples did not receive any surface treatment (control) or were coated with one of the following experimental coatings (2-hydroxyethyl methacrylate - HE; 3-hydroxypropyl methacrylate - HP; and 2-trimethylammonium ethyl methacrylate chloride - T; and sulfobetaine methacrylate - S). The concentrations of the constituent monomers were 25, 30 or 35%. The water contact angles of the samples were measured, and half of the samples were exposed to saliva. The adherent yeast cells were counted after crystal violet staining. Results: For the smooth samples, the groups S35, HP35 and HE35 showed significantly lower number of adhered Candida than control, in the absence of saliva. There were no significant differences among the experimental and control groups for the rough samples, but the saliva decreased the cell numbers for groups S25, S30 and HP30. The photoelectron spectroscopy analysis confirmed the changes in the chemical compositions of the experimental samples. Conclusions: The experimental photopolymerised coatings changed the chemical composition and decreased C. albicans adhesion in the groups S35, HP35 and HE35, suggesting that they should be further investigated. © 2012 The Gerodontology Society and John Wiley & Sons A/S.

NMDA receptors in the lateral hypothalamus have an inhibitory influence on the tachycardiac response to acute restraint stress in rats

The aim of the present study was to investigate the role of the lateral hypothalamus (LH) and its local glutamatergic neurotransmission in the cardiovascular adjustments observed when rats are submitted to acute restraint stress. Bilateral microinjection of the nonspecific synaptic inhibitor CoCl2 (0.1 nmol in 100 nL) into the LH enhanced the heart rate (HR) increase evoked by restraint stress without affecting the blood pressure increase. Local microinjection of the selective N-methyl-d-aspartate (NMDA) glutamate receptor antagonist LY235959 (2 nmol in 100 nL) into the LH caused effects that were similar to those of CoCl2. No changes were observed in the restraint-related cardiovascular response after a local microinjection of the selective non-NMDA glutamatergic receptor antagonist NBQX (2 nmol in 100 nL) into the LH. Intravenous administration of the muscarinic cholinergic receptor antagonist homatropine methyl bromide (0.2 mg/kg), a quaternary ammonium drug that does not cross the blood-brain barrier, abolished the changes in cardiovascular responses to restraint stress following LH treatment with LY235959. In summary, our findings show that the LH plays an inhibitory role on the HR increase evoked by restraint stress. Present results also indicate that local NMDA glutamate receptors, through facilitation of cardiac parasympathetic activity, mediate the LH inhibitory influence on the cardiac response to acute restraint stress. The bilateral microinjection of the CoCl2 or LY235959 into the LH enhanced the HR increase evoked by restraint stress without affecting the blood pressure increase. Intravenous administration of the homatropine methyl bromide abolished the changes in cardiovascular responses to restraint stress following LH treatment with LY235959. These results suggest that such LH influence is mediated by local NMDA glutamate receptors and involves parasympathetic nervous activation. © 2013 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

Políticas públicas e direitos humanos em Jürgen Habermas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)