Interaction of horseradish peroxidase with Langmuir monolayers of phospholipids

The method employed to incorporate guest molecules onto phospholipid Langmuir monolayers plays an important role in the interaction between the monolayer and the guest molecules. In this paper, we show that for the interaction between horseradish peroxidase (HRP) and a monolayer of dipalmitoylphosphatidylglycerol (DPPG) does depend on the method of HRP incorporation. The surface pressure isotherms of the mixed DPPG/HRP monolayers, for instance, were less expanded when the two materials were co-spread than in the case where HRP was injected into the subphase. Therefore, the method for incorporation affected not only the penetration of HRP but also the changes in molecular packing caused to the DPPG monolayer. With experiments with the monolayer on a pendant drop, we observed that the incorporation of HRP affects the dynamic elasticity of the DPPG monolayer, on a way that varies with the surface pressure. At low pressures, HRP causes the monolayer to be more rigid, while the converse is true for surface pressures above 8 mN/m. Taken all the results together, we conclude that HRP is more efficiently incorporated if injected into the subphase on which a DPPG monolayer had been spread and that the interaction between HRP and DPPG is maintained even at high surface pressures. This is promising for the possible transfer of mixed films onto solid substrates and for applications in biosensors and drug delivery systems. (c) 2008 Elsevier B.V. All rights reserved.

Interaction of oligonucleotide-based amphiphilic block copolymers with cell membrane models

Oligonucleotides have unique molecular recognition properties, being involved in biological mechanisms such as cell-surface receptor recognition or gene silencing. For their use in human therapy for drug or gene delivery, the cell membrane remains a barrier, but this can be obviated by grafting a hydrophobic tail to the oligonucleotide. Here we demonstrate that two oligonucleotides, one consisting of 12 guanosine units (G(12)), and the other one consisting of five adenosine and seven guanosine (A(5)G(7)) units, when functionalized with poly(butadiene), namely PB-G(12) and PB-A(5)G(7), can be inserted into Langmuir monolayers of dipalmitoyl phosphatidyl choline (DPPC), which served as a cell membrane model. PB-G(12) and PB-A(5)G(7) were found to affect the DPPC monolayer even at high surface pressures. The effects from PB-G(12) were consistently stronger, particularly in reducing the elasticity of the DPPC monolayers, which may have important biological implications. Multilayers of DPPC and nucleotide-based copolymers could be adsorbed onto solid supports, in the form of Y-type LB films, in which the molecular-level interaction led to lower energies in the vibrational spectra of the nucleotide-based copolymers. This successful deposition of solid films opens the way for devices to be produced which exploit the molecular recognition properties of the nucleotides. (C) 2010 Elsevier Inc. All rights reserved.

Interaction of cationic bilayer fragments with a model oligonucleotide

The interaction between cationic bilayer fragments and a model oligonucleotide was investigated by differential scanning calorimetry, turbidimetry, determination of excimer to monomer ratio of 2-(10-(1-pyrene)-decanoyl)-phosphatidyl-choline in bilayer fragment dispersions and dynamic light scattering for sizing and zeta-potential analysis. Salt (Na(2)HPO(4)), mononucleotide (2`-deoxyadenosine-5`-monophosphate) or poly (dA) oligonucleotide (3`-AAA AAA AAA A-5`) affected structure and stability of dioctadecyldimethylammonium bromide bilayer fragments. Oligonucleotide and salt increased bilayer packing due to bilayer fragment fusion. Mononucleotide did not reduce colloid stability or did not cause bilayer fragment fusion. Charge neutralization of bilayer fragments by poly (dA) at 1:10 poly (dA):dioctadecyldimethylammonium bromide molar ratio caused extensive aggregation, maximal size and zero of zeta-potential for the assemblies. Above charge neutralization, assemblies recovered colloid stability due to charge overcompensation. For bilayer fragments/poly (dA), the nonmonotonic behavior of colloid stability as a function of poly (dA) concentration was unique for the oligonucleotide and was not observed for Na(2)HPO(4) or 2`-deoxyadenosine-5`-monophosphate. For the first time, such interactions between cationic bilayer fragments and mono- or oligonucleotide were described in the literature. Bilayer fragments/oligonucleotide assemblies may find interesting applications in drug delivery. (c) 2010 Elsevier B.V. All rights reserved.

Dressing liposomal particles with chitosan and poly(vinylic alcohol) for oral vaccine delivery

Liposomes have been used as adjuvants since 1974. One major limitation for the use of liposomes in oral vaccines is the lipid structure instability caused by enzyme activities. Our aim was to combine liposomes that could encapsulate antigens (i.e., Dtxd, diphtheria toxoid) with chitosan, which protects the particles and promotes mucoadhesibility. We employed physical techniques to understand the process by which liposomes (SPC: Cho, 3: 1) can be sandwiched with chitosan (Chi) and stabilized by PVA (poly-vinylic alcohol), which are biodegradable, biocompatible polymers. Round, smooth-surfaced particles of REVs-Chi (reversed-phase vesicles sandwiched by Chi) stabilized by PVA were obtained. The REVs encapsulation efficiencies (Dtxd was used as the antigen) were directly dependent on the Chi and PVA present in the formulation. Chi adsorption on the REVs surface was accompanied by an increase of zeta-potential. In contrast, PVA adsorption on the REVs-Chi surface was accompanied by a decrease of zeta-potential. The presence of Dtxd increased the Chi surface-adsorption efficiency. The PVA affinity by mucine was 2,000 times higher than that observed with Chi alone and did not depend on the molecule being in solution or adsorbed on the liposomal surface. The liberation of encapsulated Dtxd was retarded by encapsulation within REVs-Chi-PVA. These results lead us to conclude that these new, stabilized particles were able to be adsorbed by intestinal surfaces, resisted degradation, and controlled antigen release. Therefore, REVs-Chi-PVA particles can be used as an oral delivery adjuvant.

Adolescência e infecção pelo HIV : situações de risco e proteção, autoconceito e sintomatologia psiquiátrica

Esta dissertação tem como objetivo identificar, através de um estudo transversal, associações entre as características demográficas, comportamentos de risco sexual e por drogas, sintomatologia psiquiátrica e autoconceito em uma amostra de adolescentes, comparando-os pela soropositividade HIV. Foram entrevistados 388 adolescentes entre 13 e 20 anos que procuraram espontaneamente o Centro de Triagem e Avaliação (CTA) Paulo César Bonfim para realizar o teste anti-HIV. Foram utilizados os questionários CRA (Comportamentos de Risco para AIDS), SCL-90-R (The Symptom Check-List-90-R) e EFA (Escala Fatorial de Autoconceito). A média da idade foi de 17,7 anos (±2,0). A maioria da amostra era do sexo feminino, solteiro ou sem companheiro/a fixo/a, sem trabalho, com baixas escolaridade e renda familiar. Em relação ao teste sorológico, 73% estavam realizandoo pela primeira vez, e a soropositividade geral da amostra foi de 6,2%. A escolaridade foi significativamente mais baixa em soropositivos (OR= 7,8) e não houve diferença estatística entre os grupos nas demais variáveis demográficas. Houve associação direta entre soropositividade e precocidade de início da vida sexual (OR= 3,6), história de gravidez (OR=4,7), prática de aborto prévio (OR= 5,9) e relação sexual com parceiro sem preservativo (OR=3,2). Do total da amostra, 8% dos adolescentes recebeu dinheiro em troca de relação sexual, e isto foi associado à soropositividade (OR=4,5). O mesmo ocorreu nos 13,2% que tiveram relação sexual com paceiro/a possivelmente soropositivo (OR=8,3). Apenas 18,9% da amostra afirmaram usar preservativos nos 6 meses prévios à coleta e 81,4% da amostra não haviam usado preservativo nas relações sexuais ao longo da vida. A prevalência geral de uso de drogas no mês anterior à coleta foi alta, com relato freqüente de uso de múltiplas drogas, sendo as mais freqüentes o álcool (71,6 %), maconha (24,7%), cocaína intranasal (12,4%) e solventes (9,3 %). A soropositividade foi mais alta nos usuários recentes de maconha (OR= 2,8), estimulantes (OR=2,9) e solventes (OR= 7,5). A freqüência a local específico para uso de drogas ocorreu em 22,5% do total da amostra e foi estatisticamente maior nos soropositivos (OR=2,6). Os adolescentes soropositivos apresentaram siginficativamente mais sintomatologia psiquiátrica em todas as dimensões avaliadas. Não houve diferença nas dimensões de autoconceito. O Tamanho de Efeito Padronizado foi maior nas variáveis da sintomatologia psiquiátrica do que nas do autoconceito. Os resultados indicam que os adolescentes estudados ainda não possuem cuidados preventivos suficientes para diminuir o contágio pelo vírus da AIDS. São indivíduos sob risco constante por apresentarem maior freqüência de exposição ao HIV associada à baixa escolaridade, comportamento sexual, uso de drogas e sintomatologia psiquiátrica. A falta de diferenças no autoconceito dos adolescentes soropositivos e soronegativos pode estar associada ao senso de invulnerabilidade próprio da faixa etária. Os achados apontam para a necessidade de programas específicos de prevenção para adolescentes. Medidas de intervenção futuras deverão levar em conta a interação de fatores individuais, ambientais, comportamentais e psicológicos, com o objetivo de implementar mudanças efetivas no comportamento de risco para que se detenha o avanço da epidemia nessa faixa etária.

The effect of interaction between macromolecule supplement and oxygen tension on bovine oocytes and embryos cultured in vitro

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

A violência no contexto de um serviço de urgência: análise do processo de cuidar na visão das vítimas e profissionais de saúde em Natal/RN

Exploratory descriptive study, with a quantitative approach and prospective data, performed in Pronto Socorro Clóvis Sarinho (PSCS), in Natal/RN, aiming to analyze care given by the nursing and medical staff to victims of violence attended to in an emergency hospital in Natal/RN; to identify care given by the nursing and medical staff as viewed by the victims; to compare data observed during the process of care with the victim s view on the care given by the nursing and medical staff; to identify the existing knowledge on violence and the process of caring for victims and its relation with prejudice; to identify obstacles and perspectives for prevention during the process of caring for victims in the emergency services. The population consisted of 97 physicians, 16 nurses, 75 nursing technicians and assistants and 365 victims of violence, with data collected from April to May 2009. Out of 188 professionals, 52.1% are female; 32% were aged 41 to 50; 99.5% had given care to a victim of violence; 90.4% reported to have given care to patients under custody; among these, 17.3% felt prejudice; 55.3% stated they don t provide different care for assaulted victims and assailants, however 44.7% stated they do; 86.7% feel their workplace is unsafe; 61.7% denied the existence of any obstacle and 38.3% reported the existence of obstacles; among these, 26.1% referred to inadequate facilities; 37.8% believe reinforcing security and professional training are the main solutions. Among the 365 researched violence victims, 82.2% were assaulted; male (69.6%); aged 18 to 24 (24.9%); hailing from the Greater Natal area (89.9%); on 19.7% the event happened on Saturday; during the night (48.8%); victim of physical assault (61.4%); produced by body force (27.7%); 24.4% were injured in the head and neck. 57% had used some drug, among which alcohol was predominant (75.5%). On 621 observations performed during the victim care process, when compared to the report of assaulted victims, there was a statistical difference, at 5% significance level, regarding reception, resistance from the professionals, questioning about the violent event, providing of guidance, interaction with the patient and the understanding of receiving proper care, and care resolution. In comparisons involving the observed and the assailant victims reports, there was a statistical difference regarding the resence of resistance from the professionals, performance of necessary procedures and the nteraction with the patient and the understanding of receiving proper care and 58.1% reported the nursing team was the one that provided the best care. We conclude that professionals had lready given care to assailant patients, acknowledge the importance of knowing how the vent took place and acquired this preparation during their practice. The most often referred bstacles that hinder assistance were: inadequate facilities, material deficit and lack of rofessional preparation. As solutions for these problems, they cited the reinforcement of ecurity and professional training

Study of the interaction between cardiolipin bilayers and methylene blue in polymer-based Layer-by-Layer and Langmuir films applied as membrane mimetic systems

An increase of the reports involving mimetic systems has been observed. Briefly, these systems use biological phospholipids to exploit specific interactions between membrane-models and drugs. Here, the Layer-by-Layer (LbL) and Langmuir techniques were used to investigate the interaction between cardiolipin (CLP-negative phospholipid) and a cationic-like drug methylene blue (MB). Supported by a cationic polyelectrolyte (PAH), LbL films containing PAH/(CLP + MB) and PAH/(CLP + MB + AgNP) were grown up to 14 bilayers. The optical microscopy analysis revealed a decrease of the CLP vesicle sizes in the presence of MB as a possible consequence of the MB action onto the mechanical properties of the CLP membrane. From FTIR spectra, changes mainly related to peak position and band intensity and shape were observed in the spectra from PAH/CLP when in the presence of MB. The latter supports that the interactions between the phosphate and amine charged groups from CLP and PAH, respectively, established during the LbL film fabrication, besides the CLP hydrocarbon environment, are influenced by the presence of MB. Using the micro-Raman technique, a chemical mapping was build based on MB spectrum by resonance Raman scattering (RRS) and surface-enhanced resonance Raman scattering (SERRS). The later phenomenon was activated by Ag nanoparticles (AgNPs) trapped within the LbL film allowing collecting spectra for a single bilayer of PAH/(CLP + MB + AgNP). A rough estimation showed a SERRS amplification of 10(3) in comparison to RRS spectra. As a complementary approach, Langmuir films of CLP in the presence of co-spread MB were investigated through surface pressure vs mean molecular area (pi-A) isotherms. The results showed that for concentrations of MB below 100 mol%, the drug is expelled to water subphase for high values of surface pressure (condensed phase). For concentration at 100% and higher, the MB keeps bound to CLP floating monolayer. (C) 2010 Elsevier B.V. All rights reserved.

Maternal separation stress in male mice: long-term increases in alcohol intake

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Alcohol use disorders in patients with obsessive-compulsive disorder: The importance of appropriate dual-diagnosis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Interaction of shikimic acid with shikimate kinase (Retracted Article. See vol 334, pg 967, 2005)

The crystal structure of shikimate kinase from Mycobacterium tuberculosis (MtSK) complexed with MgADP and shikimic acid (shikimate) has been determined at 2.3 Angstrom resolution, clearly revealing the amino acid residues involved in shikimate binding. In MtSK, the Glu61 strictly conserved in SK forms a hydrogen bond and salt-bridge with Arg58 and assists in positioning the guanidinium group of Arg58 for shikimate binding. The carboxyl group of shikimate interacts with Arg58, Gly81, and Arg136, and hydroxyl groups with Asp34 and Gly80. The crystal structure of MtSK-MgADP-shikimate will provide crucial information for elucidation of the mechanism of SK-catalyzed reaction and for the development of a new generation of drugs against tuberculosis. (C) 2004 Elsevier B.V. All rights reserved.

Synthesis and Characterization of Novel, Highly Crystalline Poly(vinyl alcohol) Microspheres for Chemoembolization Therapy

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

Interaction between paraventricular nucleus and septal area in the control of physiological responses induced by angiotensin II

We determined the effects of losartan (40 nmol) and PD 123319 (40 nmol) (both non-peptides and selective antagonists of the AT1 and AT2 angiotensin receptors, respectively), and [Sar¹, Ala8] angiotensin II (ANG II) (40 nmol) (a non-selective peptide antagonist of angiotensin receptors) injected into the paraventricular nucleus (PVN) on the water and salt appetite, diuresis and natriuresis and mean arterial pressure (MAP) induced by administration of 10 nmol of ANG II into the medial septal area (MSA) of male Holtzman rats weighing 250-300 g. The volume of drug solution injected was 0.5 µl over a period of 10-15 s. The responses were measured over a period of 120 min. ANG II alone injected into the MSA induced an increase in all the above parameters (8.1 ± 1.2, 1.8 ± 0.3, and 17.1 ± 1.0 ml, 217 ± 25 µEq/120 min, and 24 ± 4 mmHg, respectively, N = 10-12) compared with vehicle-treated rats (1.4 ± 0.2, 0.6 ± 0.1, and 9.3 ± 0.5 ml, 47 ± 5 µEq/120 min, and 4.1 ± 0.8 mmHg, respectively, N = 10-14). Pretreatment with losartan and [Sar¹, Ala8] ANG II completely abolished the water and sodium intake, and the pressor increase (0.5 ± 0.2, 1.1 ± 0.2, 0.5 ± 0.2, and 0.8 ± 0.2 ml, and 1.2 ± 3.9, 31 ± 4.6 mmHg, respectively, N = 9-12), whereas losartan blunted the urinary and sodium excretion induced by ANG II (13.9 ± 1.0 ml and 187 ± 10 µEq/120 min, respectively, N = 9). Pretreatment with PD 123319 and [Sar¹, Ala8] ANG II blocked the urinary and sodium excretion (10.7 ± 0.8, 9.8 ± 0.7 ml, and 67 ± 13 and 57 ± 17 µEq/120 min, respectively, N = 9), whereas pretreatment with PD 123319 partially blocked the water and sodium intake, and the MAP induced by ANG II administration (2.3 ± 0.3, 1.1 ± 0.1 ml, and 12 ± 3 mmHg, respectively, N = 9-10). These results suggest the angiotensinergic effect of the MSA on the AT1 and AT2 receptors of the PVN in terms of water and sodium homeostasis and MAP modulation.

Structural Basis for Interaction of Inhibitors with Cyclin-Dependent Kinase 2

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

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.