Translocating the blood-brain barrier using electrostatics

Copyright © 2012 Ribeiro,Domingues, Freire,Santos and Castanho. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.Mammalian cell membranes regulate homeostasis, protein activity, and cell signaling. The charge at the membrane surface has been correlated with these key events. Although mammalian cells are known to be slightly anionic, quantitative information on the membrane charge and the importance of electrostatic interactions in pharmacokinetics and pharmacodynamics remain elusive. Recently, we reported for the first time that brain endothelial cells (EC) are more negatively charged than human umbilical cord cells, using zeta-potential measurements by dynamic light scattering. Here, we hypothesize that anionicity is a key feature of the blood-brain barrier (BBB) and contributes to select which compounds cross into the brain. For the sake of comparison, we also studied the membrane surface charge of blood components—red blood cells (RBC), platelets, and peripheral blood mononuclear cells (PBMC).To further quantitatively correlate the negative zeta-potential values with membrane charge density, model membranes with different percentages of anionic lipids were also evaluated. From all the cells tested, brain cell membranes are the most anionic and those having their lipids mostly exposed, which explains why lipophilic cationic compounds are more prone to cross the blood-brain barrier.Fundação para a Ciência e Tecnologia — Ministério da Educação e Ciência (FCT-MEC, Portugal) is acknowledged for funding (including fellowships SFRH/BD/42158/2007 to Marta M.B. Ribeiro, SFRH/BD/41750/2007 to Marco M. Domingues and SFRH/BD/70423/2010 to João M. Freire) and project PTDC/QUI-BIQ/119509/2010. Marie Curie Industry-Academia Partnerships and Pathways (European Commission) is also acknowledged for funding (FP7-PEOPLE-2007-3-1-IAPP, Project 230654)

Electronic doping of graphene by deposited transition metal atoms

We perform a phenomenological analysis of the problem of the electronic doping of a graphene sheet by deposited transition metal atoms, which aggregate in clusters. The sample is placed in a capacitor device such that the electronic doping of graphene can be varied by the application of a gate voltage and such that transport measurements can be performed via the application of a (much smaller) voltage along the graphene sample, as reported in the work of Pi et al. [Phys. Rev. B 80, 075406 (2009)]. The analysis allows us to explain the thermodynamic properties of the device, such as the level of doping of graphene and the ionisation potential of the metal clusters in terms of the chemical interaction between graphene and the clusters. We are also able, by modelling the metallic clusters as perfect conducting spheres, to determine the scattering potential due to these clusters on the electronic carriers of graphene and hence the contribution of these clusters to the resistivity of the sample. The model presented is able to explain the measurements performed by Pi et al. on Pt-covered graphene samples at the lowest metallic coverages measured and we also present a theoretical argument based on the above model that explains why significant deviations from such a theory are observed at higher levels of coverage.Comment: 16 pages, 10 figure

Linking cardiorespiratory fitness classification criteria to early subclinical atherosclerosis in children

It is unclear if cardiorespiratory fitness (CRF) can be used as a screening tool for premature changes in carotid intima-media thickness (cIMT) in paediatric populations. The purpose of this cross-sectional study was 3-fold: (i) to determine if CRF can be used to screen increased cIMT; (ii) to determine an optimal CRF cut-off to predict increased cIMT; and (iii) to evaluate its ability to predict increased cIMT among children in comparison with existent CRF cut-offs. cIMT was assessed with high-resolution ultrasonography and CRF was determined using a maximal cycle test. Receiver operating characteristic analyses were conducted in boys (n = 211) and girls (n = 202) aged 11-12 years to define the optimal sex-specific CRF cut-off to classify increased cIMT (≥75th percentile). Logistic regression was used to examine the association between the CRF cut-offs with the risk of having an increased cIMT. The optimal CRF cut-offs to predict increased cIMT were 45.81 and 34.46 mL·kg(-1)·min(-1) for boys and girls, respectively. The odds-ratios for having increased cIMT among children who were unfit was up to 2.8 times the odds among those who were fit (95% confidence interval: 1.40-5.53). Considering current CRF cut-offs, only those suggested by Adegboye et al. 2011. (Br. J. Sports Med. 45(9): 722-728) and Boddy et al. 2012 (PLoS One, 7(9): e45755) were significant in predicting increased cIMT. In conclusion, CRF cut-offs (boys: ≤ 45.8; girls: ≤ 34.5 mL·kg(-1)·min(-1)) are associated with thickening of the arterial wall in 11- to 12-year-old children. Low CRF is an important cardiovascular risk factor in children and our data highlight the importance of obtaining an adequate CRF.info:eu-repo/semantics/publishedVersio

HIV-1 Fusion Inhibitor Peptides Enfuvirtide and T-1249 Interact with Erythrocyte and Lymphocyte Membranes

Enfuvirtide and T-1249 are two HIV-1 fusion inhibitor peptides that bind to gp41 and prevent its fusogenic conformation, inhibiting viral entry into host cells. Previous studies established the relative preferences of these peptides for membrane model systems of defined lipid compositions. We aimed to understand the interaction of these peptides with the membranes of erythrocytes and peripheral blood mononuclear cells. The peptide behavior toward cell membranes was followed by di-8-ANEPPS fluorescence, a lipophilic probe sensitive to the changes in membrane dipole potential. We observed a fusion inhibitor concentration-dependent decrease on the membrane dipole potential. Quantitative analysis showed that T-1249 has an approximately eight-fold higher affinity towards cells, when compared with enfuvirtide. We also compared the binding towards di-8-ANEPPS labeled lipid vesicles that model cell membranes and obtained concordant results. We demonstrated the distinct enfuvirtide and T-1249 membranotropism for circulating blood cells, which can be translated to a feasible in vivo scenario. The enhanced interaction of T-1249 with cell membranes correlates with its higher efficacy, as it can increase and accelerate the drug binding to gp41 in its pre-fusion state

rBPI21 Promotes Lipopolysaccharide Aggregation and Exerts Its Antimicrobial Effects by (Hemi)fusion of PG-Containing Membranes

Antimicrobial peptides (AMPs) are important potential alternatives to conventional therapies against bacterial infections. rBPI21 is a 21 kDa peptide based on the N-terminal region of the neutrophil bactericidal/permeability-increasing protein (BPI). This AMP possesses highly selective bactericidal effects on Gram-negative bacteria and have affinity for lipopolysaccharide (LPS) which is believed to be at the origin of its neutralizing effect of the LPS segregated into the bloodstream. We aim at understanding the molecular bases of rBPI21 bactericidal and LPS neutralization actions, using biomembrane model systems. Using dynamic light scattering spectroscopy we demonstrate that rBPI21 promotes aggregation of negatively charged large unilamellar vesicles (LUV), even in the absence of LPS, and LPS aggregates, while for zwitterionic phosphatidylcholine (POPC) LUV the size remains unchanged. The peptide also promotes the fusion (or hemifusion) of membranes containing phosphatidylglycerol (POPG). The aggregation and fusion of negatively charged LUV are peptide concentration-dependent until massive aggregation is reached, followed by sample flocculation/precipitation. Concomitantly, there is a progressive change in the zeta-potential of the LUV systems and LPS aggregates. LUV systems composed of phosphatidylglycerol (POPG) and lipid mixtures with POPG have higher zeta-potential variations than in the absence of POPG. The interaction of rBPI21 with lipid vesicles is followed by leakage, with higher effect in POPG-containing membranes. LPS aggregation can be related with a decreased toxicity, possibly by facilitating its clearance by macrophage phagocytosis and/or blocking of LPS specific receptor recognition. Our data indicate that rBPI21 mechanism of action at the molecular level involves the interaction with the LPS of the outer membrane of Gram-negative bacteria, followed by internalization and leakage induction through the (hemi)fusion of the bacterial outer and inner membranes, both enriched in phosphatidylglycerol

The effect of the interproximal creeping attachment in aesthetic site after root coverage surgery with VISTA technique - a case report

Gingival recession is a common manifestation in most populations. The mechanism by which gingival recession occurs is not well understood, but it seems to be complex and multifactorial. The main etiological factors are the accumulation of dental plaque biofilm with the resulting inflammatory periodontal diseases and mechanical trauma due to faulty oral hygiene techniques, especially in thin biotypes. This case report describes the treatment of a vestibular recession associated with interdental bone loss, with the VISTA technique associated with a connective tissue graft. The case was evaluated at 3, 9 months and 48 months after the surgery clinically complete root coverage and increased thickness of keratinized tissue were achieved, and the interdental papilla was augmented improving the soft tissue quality for future orthodontic treatment. VISTA technique associated with a connective tissue graft to reconstruct vertically papilla is a promising alternative for minimally invasive treatment and stable after 4 years.info:eu-repo/semantics/publishedVersio

Evolution of squeezed states under the Fock-Darwin Hamiltonian

We develop a complete analytical description of the time evolution of squeezed states of a charged particle under the Fock-Darwin Hamiltonian and a time-dependent electric field. This result generalises a relation obtained by Infeld and Pleba\'nski for states of the one-dimensional harmonic oscillator. We relate the evolution of a state-vector subjected to squeezing to that of state which is not subjected to squeezing and for which the time-evolution under the simple harmonic oscillator dynamics is known (e.g. an eigenstate of the Hamiltonian). A corresponding relation is also established for the Wigner functions of the states, in view of their utility in the analysis of cold-ion experiments. In an appendix, we compute the response functions of the FD Hamiltonian to an external electric field, using the same techniques as in the main text

Conjugation of cholesterol to HIV-1 fusion inhibitor C34 increases peptide-membrane interactions potentiating its action

© 2013 Hollmann et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Recently, the covalent binding of a cholesterol moiety to a classical HIV-1 fusion inhibitor peptide, C34, was shown to potentiate its antiviral activity. Our purpose was to evaluate the interaction of cholesterol-conjugated and native C34 with membrane model systems and human blood cells to understand the effects of this derivatization. Lipid vesicles and monolayers with defined compositions were used as model membranes. C34-cholesterol partitions more to fluid phase membranes that mimic biological membranes. Importantly, there is a preference of the conjugate for liquid ordered membranes, rich in cholesterol and/or sphingomyelin, as observed both from partition and surface pressure studies. In human erythrocytes and peripheral blood mononuclear cells (PBMC), C34-cholesterol significantly decreases the membrane dipole potential. In PBMC, the conjugate was 14- and 115-fold more membranotropic than T-1249 and enfuvirtide, respectively. C34 or cholesterol alone did not show significant membrane activity. The enhanced interaction of C34-cholesterol with biological membranes correlates with its higher antiviral potency. Higher partitions for lipid-raft like compositions direct the drug to the receptor-rich domains where membrane fusion is likely to occur. This intermediary membrane binding step may facilitate the drug delivery to gp41 in its pre-fusion state.This work was funded by Fundação para a Ciência e Tecnologia, Portugal (fellowships SFRH/BPD/72037/2010 and SFRH/BD/42205/2007 to A.H. and P.M.M., respectively, and projects PTDC/QUI-BIQ/104787/2008, PTDC/QUI-BIQ/112929/2009 and VIH/SAU/0047/2011)

Black holes with synchronised Proca hair: linear clouds and fundamental non-linear solutions

Recent studies have made key progress on the black hole/solitonic solutions of the Einstein-Proca system. Firstly, fully non-linear dynamical evolutions of the Kerr black hole superradiant instability, triggered by a Proca field, have shown the formation of a new equilibrium state, a spinning black hole with synchronised Proca hair. Secondly, non-linear evolutions of spinning Proca stars have established that they are dynamically stable, unlike their scalar cousins. Thirdy, separability of the Proca equation on the Kerr background has been achieved. Motivated by these results, in this paper we reconsider Kerr black holes with synchronised Proca hair. The separability of the Proca equation on the Kerr background allows us to examine the stationary Proca clouds in greater detail, in particular their dependence on the different quantum numbers. These stationary clouds occur at a set of existence lines in the Kerr parameter space, from which the black holes with synchronised Proca hair bifurcate. We construct the domain of existence of these black holes, comparing the fundamental states missed in the original study with the first excited states and with the cousin scalar model, giving illustrative examples of Kerr-like and non-Kerr-like BHs. In the vanishing event horizon limit, these hairy black holes connect to the fundamental states of spinning Proca stars, which include the dynamically stable solutions.Comment: 36 pages, 13 figure

Using zeta-potential measurements to quantify peptide partition to lipid membranes

© The Author(s) 2011. This article is published with open access at Springerlink.com.Open Access: This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.Many cellular phenomena occur on the biomembranes. There are plenty of molecules (natural or xenobiotics) that interact directly or partially with the cell membrane. Biomolecules, such as several peptides (e.g., antimicrobial peptides) and proteins, exert their effects at the cell membrane level. This feature makes necessary investigating their interactions with lipids to clarify their mechanisms of action and side effects necessary. The determination of molecular lipid/water partition constants (Kp) is frequently used to quantify the extension of the interaction. The determination of this parameter has been achieved by using different methodologies, such as UV-Vis absorption spectrophotometry, fluorescence spectroscopy and ζ-potential measurements. In this work, we derived and tested a mathematical model to determine the Kp from ζ-potential data. The values obtained with this method were compared with those obtained by fluorescence spectroscopy, which is a regular technique used to quantify the interaction of intrinsically fluorescent peptides with selected biomembrane model systems. Two antimicrobial peptides (BP100 and pepR) were evaluated by this new method. The results obtained by this new methodology show that ζ-potential is a powerful technique to quantify peptide/lipid interactions of a wide variety of charged molecules, overcoming some of the limitations inherent to other techniques, such as the need for fluorescent labeling.This work was partially supported by project PTDC/QUI/ 69937/2006 from Fundação para a Ciência e Tecnologia-Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES, Portugal), and by Fundação Calouste Gulbenkian (Portugal). JMF and MMD also thank FCT-MCTES for grants IMM/BT/37-2010 and SFRH/BD/41750/2007, respectively