Search

Made available in DSpace on 2021-06-25T10:53:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) EMC Dengue virus infection depends on its fusion with the host membrane, where the binding occurs through interaction between proteins on the virus cell surface and specific viral receptors on target membranes. This process is mediated by the fusion peptide located between residues 98 and 112 (DRGWGNGCGLFGKGG) that forms a loop in domain II of dengue E glycoprotein. In this study, we evaluated the role of fusion peptide surrounding regions (88–97 and 113–123) of the Dengue 2 subtype on its interaction with the membrane and fusion activity. These sequences are important to stabilize the fusion peptide loop and increase fusion activity. Three peptides, besides the fusion peptide, were synthesized by SPPS using the Fmoc chemical approach. The first contains the fusion peptide and the C-terminal region of the loop (sequence 98–123); another contains the N-terminal region (88–112) and the larger peptide contains both regions (88–123). The peptides were able to interact with a model membrane. Differences in morphology of the monolayer promoted by the peptides were assessed by Brewster Angle Microscopy (BAM). Our data indicated that the C-terminal region of fusion peptide loop is more efficient in promoting fusion and interacting with the membrane than the N-terminal sequence, which is responsible for the electrostatic initial interaction. We propose a 2-step mechanism for the interaction of the dengue virus fusion peptide with the host membrane, where the N-terminal sequence docks electrostatically on the headgroups and then the C-terminal interacts via hydrophobic forces in the acyl chains. Departamento de Bioquímica e Química Orgânica Instituto de Química UNESP -Univ Estadual Paulista Instituto de Física de São Carlos Universidade de São Paulo USP, São Carlos Department of Chemistry The Ohio State University Departamento de Bioquímica e Química Orgânica Instituto de Química UNESP -Univ Estadual Paulista

The effect of temperature on stability of lipid microbubble shell containing polyethyleneoxide-40-stearate (PEG40St) as emulsifier was investigated. Microbubbles at 4 °C were subjected to different temperatures up to 48 ºC (down-to-up) and it was found that both the number and the size of microbubbles remained unchanged in the population up to a certain time, so called “onset time”. The onset time was about 6 hours at 10 °C, 2 hours at 20 °C and shorter at elevated temperatures, exhibiting an exponential decrease with increasing temperature. Once the onset time was reached, the number of microbubbles started to decrease and the average size of the population started to increase. Observation of single microbubbles on a constant temperature heating stage exhibited that each microbubble had its own onset time, with the smaller microbubbles vanishing earlier than the larger ones. The Langmuir monolayer studies showed that hydration degree of the emulsifier PEG chains decreased with temperature, causing them go through conformational changes and subsequently destabilization of the shell. By subjecting the freshly produced microbubbles directly to the desired temperatures in up-to-down fashion, more stable microbubbles were able to be produced, with their onset time increased 40% at 10 °C to 500% at 38 °C. Overall, the results suggest that the new strategies need to be developed to control the collapse process in the microbubble shell resulting from the conformational changes in the PEG chains of the emulsifier for the design of more stable microbubbles. © 2019, Turkish Chemical Society. All rights reserved.

Made available in DSpace on 2019-10-04T12:15:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Flavonoid-metal complexes are widely studied because of their interesting luminescent behavior and biological activity. Despite the extensive exploration of flavonoid-metal coordination processes in solution, the formation of complexes using the flavonoid molecule inserted in a lipid membrane has been little investigated. This effect could provide important insight into the biological activity of flavonoids at lipid membranes and could represent an attractive strategy to design supramolecular structures. Here, we studied the complexation between Sr2+ and morin inserted in an octadecylphosphonic acid (OPA) Langmuir monolayer. This is a relevant system due to the synergism imposed by the association of the Sr2+ ability to control bone formation/resorption with the morin antioxidative effect. Morin incorporation into the OPA monolayers and further Sr2+ complexation were monitored by surface pressure isotherms. Electronic absorption spectroscopy and fluorescence techniques showed Srmorin complexation both in solution and at the air-liquid interface. Although morin complexation has been described to occur only at basic pH, the specific thermodynamic properties at the air-liquid interface drove metal complexation. LB films were deposited on Ti surfaces, and the resulting OPA/Sr-morin coatings exhibited high surface free energy and increase on its polar component. This optimized surface feature supported further serum protein adsorption and osteoblast growth and differentiation, indicating that these lipid-based coatings are promising for bioactive coating design. This study paves the way for the use of this lipid-based coating in the design of implants for faster osteointegration. Moreover, flavonoid-metal complexation at membranes could also help to shed light on the biological role played by flavonoids. Univ Sao Paulo, Dept Quim, Fac Filosofia Ciencias & Letras Ribeirao Preto, Sao Paulo, Brazil Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil Univ Sao Paulo, Dept Fis, Fac Filosofia Ciencias & Letras Ribeirao Preto, Sao Paulo, Brazil Univ Estadual Paulista, Dept Fis, Fac Ciencias & Tecnol, Sao Paulo, Brazil Univ Estadual Paulista, Dept Fis, Fac Ciencias & Tecnol, Sao Paulo, Brazil FAPESP: 2015/08774-0 FAPESP: 2017/08892-9 FAPESP: 2016/21236-0

Understanding the interaction of ions with organic receptors in confined space is of fundamental importance and could advance nanoelectronics and sensor design. In this work, metal ion complexation of conformationally varied thiacalix[4]monocrowns bearing lower-rim hydroxy (type I), dodecyloxy (type II), or methoxy (type III) fragments was evaluated. At the liquid–liquid interface, alkylated thiacalixcrowns-5(6) selectively extract alkali metal ions according to the induced-fit concept, whereas crown-4 receptors were ineffective due to distortion of the crown-ether cavity, as predicted by quantum-chemical calculations. In type-I ligands, alkali-metal ion extraction by the solvent-accessible crown-ether cavity was prevented, which resulted in competitive Ag+ extraction by sulfide bridges. Surprisingly, amphiphilic type-I/II conjugates moderately extracted other metal ions, which was attributed to calixarene aggregation in salt aqueous phase and supported by dynamic light scattering measurements. Cation–monolayer interactions at the air–water interface were monitored by surface pressure/potential measurements and UV/visible reflection–absorption spectroscopy. Topology-varied selectivity was evidenced, towards Sr2+ (crown-4), K+ (crown-5), and Ag+ (crown-6) in type-I receptors and Na+ (crown-4), Ca2+ (crown-5), and Cs+ (crown-6) in type-II receptors. Nuclear magnetic resonance and electronic absorption spectroscopy revealed exocyclic coordination in type-I ligands and cation–π interactions in type-II ligands.

One of the main known effects of cholesterol is to rigidify the cell membrane throughout the so-called condensing effect. Although many studies have been done in mixtures of cholesterol with different membrane lipids, there are not many studies in a wide concentration range of cholesterol or at physiological conditions. In this work, we studied mixtures of DMPC/Cholesterol monolayers to determine the effect of cholesterol, from very low to physiological concentrations and two pHs. We use a Langmuir balance and Brewster angle microscopy to study their thermodynamic behavior at 37.0 ± 0.1°C at the air/solution interface. From the analysis of the (π−A) isotherms, we determined the excess area and the compressibility elastic modulus to determine the monolayers mechanical properties. Surprisingly, we found three main effects of cholesterol: The first one is a fluidization effect of the monolayer at all cholesterol concentrations. The second effect is the so-called condensing effect that appears due to the non-ideality of the mixture. The third effect is a stiffness of the monolayer as the cholesterol concentration increases. These effects are stronger in pure water, pH ≈ 6.6, than on buffer at physiological pH = 7.4. We also found that all mixtures are thermodynamically stable at all concentrations at a surface pressure of 30.1 ± 1.6 and 27.4 ± 3.2 mN/m in pure water and buffer, respectively. Furthermore, we compared this stability with a fatty acid monolayer that shows a much lower surface pressure equilibrium value that DMPC or its mixtures with cholesterol, indicating a possibly reason why double chain lipids are better than single chain lipids to made up the cell membrane.

Langmuir and Langmuir-Blodgett films holding a synthetic bioinspired wound healing active compound were used as drug-delivery platforms. Palmitic acid Langmuir monolayers were able to incorporate 2-methyltriclisine, a synthetic Triclisine derivative that showed wound healing activity. The layers proved to be stable and the nanocomposites were transferred to solid substrates. Normal human lung cells (Medical Research Council cell strain 5, MRC-5) were grown over the monomolecular Langmuir-Blodgett films that acted as a drug reservoir and delivery system. The proliferation and migration of the cells were clearly affected by the presence of 2-methyltriclisine in the amphiphilic layers. The methodology is proposed as a simple and reliable model for the study of the effects of bioactive compounds over cellular cultures., Langmuir monolayers, Drug-carrier composites, Cell culture, Triclisine

Three-finger toxins are naturally occurring proteins in Elapidae snake venoms. Nowadays, they are gaining popularity because of their therapeutic potential. On the other hand, these proteins may cause undesirable reactions inside the body′s cells. A full assessment of the safety of Naja ashei venom components for human cell application is still unknown. The aim of the study was to determine the effect of the exogenous application of three-finger toxins on the cells of monocytes (U-937) and promyelocytes (HL-60), with particular emphasis on the modification of their membranes under the influence of various doses of 3FTx protein fraction (0–120 ng/mL). The fraction exhibiting the highest proportion of 3FTx proteins after size exclusion chromatography (SEC) separation was used in the experiments. The structural response of cell membranes was described on the basis of single-component and multi-component Langmuir monolayers that mimicked the native membranes. The results show that the mechanism of protein–lipid interactions depends on both the presence of lipid polar parts (especially zwitterionic type of lipids) and the degree of membrane saturation (the greatest-for unsaturated lipids). The biochemical indicators reflecting the tested cells (MDA, LDH, cell survival, induction of inflammation, LD50) proved the results that were obtained for the model.

Made available in DSpace on 2021-06-25T11:15:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Nacional de Ciência e Tecnologia em Eletrônica Orgânica Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The lack of studies involving the effects in human health associated with the chronic ingestion of pollutants lead to the path of investigating the action of these compounds in cell membrane models. We demonstrated the interaction (causes and consequences) of the hormone 17 α-ethinylestradiol (EE2) with lipid monolayers (prepared as Langmuir films) and bilayers prepared as small unilamellar vesicles (SUVs) and giant unilamellar vesicles (GUVs). Both fluidity and majority chemical composition of real plasma cell membrane were guaranteed using the phospholipid 1-palmitoil-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC). Surface pressure-mean molecular area (π-A) isotherms and PM-IRRAS measurements highlighted the strong interaction of EE2 with POPC monolayers, leading the hormone to remain at the air/water interface and promoting its penetration into the phospholipid hydrophobic chains. In the case of bilayers, the entrance of the hormone inside the SUV is likely facilitated by their high curvature. In GUVs, EE2 was responsible for changes in the spherical shape, forming structures like buds and lipid protrusions. The set of results indicates the strong effects of EE2 on fluid membranes, which is an important feature to predict its damage in human cells. Department of Physics School of Technology and Applied Sciences São Paulo State University (UNESP) Institute of Sciences of Food Production Italian National Research Council (CNR-ISPA), S.P. Lecce-Monteroni Department of Biological and Environmental Sciences and Technologies (DiSTeBA) University of Salento S.P. Lecce-Monteroni Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM) Unità di Lecce, S.P. Lecce-Monteroni Department of Physics School of Technology and Applied Sciences São Paulo State University (UNESP) FAPESP: 2016/09633-4 FAPESP: 2018/222147-6 CAPES: 88882.330129/2018-01

Made available in DSpace on 2021-06-25T10:28:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The inadequate disposal and the difficulty in its removal from water treatment systems have made the endocrine disruptor bisphenol A (BPA) a significant hazard for humans and animals. The molecular-level mechanisms of BPA action are not known in detail, which calls for systematic investigations using cell membrane models. This paper shows that BPA affects Langmuir monolayers and giant unilamellar vesicles (GUVs) of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) used as membrane models, in a concentration-dependent manner and with effects that depend on BPA aggregation. BPA increases DPPC monolayer fluidity in surface pressure isotherms upon interacting with the headgroups through hydrogen bonding, according to polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS). In DPPC GUVs, BPA induced wrinkling and distortion in the spherical shape of the vesicles, but this was only observed for fresh solutions where it is not aggregated. BPA also decreased the viability of hamster ovary cells (CHO) in in vitro experiments. In contrast, aged, aggregated BPA solutions did not affect the GUVs and even increased CHO viability. These results may be rationalized in terms of size-dependent effects of BPA, which may be relevant for its endocrine-disrupting effects. São Paulo State University (UNESP) School of Technology and Applied Sciences São Carlos Institute of Physics University of São Paulo, CP 369 São Paulo State University (UNESP) School of Technology and Applied Sciences CNPq: 304836/2018-4 CNPq: 422163/2018-0

Human bestrophin-1 protein (hBest1) is a transmembrane channel associated with the calcium-dependent transport of chloride ions in the retinal pigment epithelium as well as with the transport of glutamate and GABA in nerve cells. Interactions between hBest1, sphingomyelins, phosphatidylcholines and cholesterol are crucial for hBest1 association with cell membrane domains and its biological functions. As cholesterol plays a key role in the formation of lipid rafts, motional ordering of lipids and modeling/remodeling of the lateral membrane structure, we examined the effect of different cholesterol concentrations on the surface tension of hBest1/POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and hBest1/SM Langmuir monolayers in the presence/absence of Ca2+ ions using surface pressure measurements and Brewster angle microscopy studies. Here, we report that cholesterol: (1) has negligible condensing effect on pure hBest1 monolayers detected mainly in the presence of Ca2+ ions, and, (2) induces a condensing effect on composite hBest1/POPC and hBest1/SM monolayers. These results offer evidence for the significance of intermolecular protein&ndash, lipid interactions for the conformational dynamics of hBest1 and its biological functions as multimeric ion channel.

Fipronil is a widely used commercial insecticide whose action mechanism consists in blocking the influx of chloride ions through the γ-aminobutyric acid type A receptor (GABAA-R), an integral membrane protein. The present study investigates the interaction of fipronil with phospholipid Langmuir monolayers, in order to characterize the effects that its partition could exert on the physical properties of these model membranes. A combined experimental and molecular dynamics (MD) simulations approach was performed. MD simulations were conducted in such a way that they resemble an experimental compression isotherm of DPPC in the presence of fipronil in the aqueous subphase. Both the experimental and the simulated compression isotherm showed that the partition of fipronil between DPPC molecules induces an expansion of the monolayer. Experimental results also showed that fipronil can penetrate lipid monolayers even in condensed packing states. MD simulations showed that fipronil induces an ordering effect in the acyl chains of DPPC in the liquid-condensed phase. In addition, the simulations indicate that fipronil orients parallel to the plane of the monolayer and that it establishes hydrogen bonds with the glycerol region of DPPC. Free energy profiles of the partition of fipronil into the monolayers, obtained by means of umbrella sampling, indicated that its penetration is thermodynamically favorable, being the interphase between the glycerol region and the acyl chains of DPPC its most favorable location. Our results suggest that fipronil could modulate the supramolecular organization of biological membranes surrounding GABAA-R, contributing, at least in part, to its action mechanism. Fil: Felsztyna, Iván. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Sánchez Borzone, Mariela E.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Miguel, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina Fil: Garcia, Daniel Asmed. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina

Submitted by Mirella Boaro Kobal (mirella.kobal@unesp.br) on 2021-02-24T03:02:20Z No. of bitstreams: 1 Sabrina_revised version_june_2020.docx: 1687573 bytes, checksum: 6cf2a10c103f27a85a34f8dcb1b03e57 (MD5) Rejected by Maria Luiza Carpi Semeghini (luiza@assis.unesp.br), reason: De acordo com a política desta revista, a versão do artigo a ser incluída no Repositório Institucional deve ser a versão aceita pela revista (postprint). Assim, o arquivo enviado em versão preprint deverá ser substituído pela versão postprint. Agradecemos a compreensão. Atenciosamente, Equipe do Repositório Unesp on 2021-02-25T18:40:58Z (GMT) Submitted by Mirella Boaro Kobal (mirella.kobal@unesp.br) on 2021-03-02T20:36:36Z No. of bitstreams: 1 Sabrina_revised version_june_2020.pdf: 1448731 bytes, checksum: a3d600b9a64085eae9442185c44611bb (MD5) Approved for entry into archive by Maria Luiza Carpi Semeghini (luiza@assis.unesp.br) on 2021-03-08T13:09:42Z (GMT) No. of bitstreams: 1 camacho_sa_postprint_assis_molecular.pdf: 1448731 bytes, checksum: a3d600b9a64085eae9442185c44611bb (MD5) Made available in DSpace on 2021-03-08T13:09:42Z (GMT). No. of bitstreams: 1 camacho_sa_postprint_assis_molecular.pdf: 1448731 bytes, checksum: a3d600b9a64085eae9442185c44611bb (MD5) Previous issue date: 2020-10 Outra Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Nanopartículas metálicas são agentes promissores para terapia fototérmica (TFT) em cânceres devido à sua fotoestabilidade e conversão eficiente de luz em calor, mas sua possível agregação continua sendo um problema. Neste artigo, relatamos o aquecimento fotoinduzido de nanopartículas de ouro recobertas com sílica (AuSHINs) em experimentos in vitro com células humanas de carcinoma de orofaringe (HEp-2) e de mama (BT-474 e MCF-7), com redução da viabilidade celular abaixo de 50% com 2,2 × 1012 AuSHINs / mL e 6 h de incubação. Esta toxicidade para as células cancerosas é significativamente maior do que em trabalhos anteriores com nanopartículas de ouro. Considerando as dimensões das AuSHINs, hipotetizamos que a absorção celular não é simples, e o mecanismo de ação envolve o acúmulo nas membranas fosfolipídicas, sendo o alvo da TFT para aquecimento fotoinduzido e geração subsequente de espécies reativas de oxigênio (ROS). Usando monocamadas de Langmuir como modelos de membrana simplificados, confirmamos que AuSHINs têm um efeito maior em 1,2-dioleoil-snglicero-3-fosfo-L-serina (DOPS), que se acredita representar as membranas de células cancerosas, do que em 1,2-dioleoil-sn -glicero-3-fosfocolina (DOPC) considerada como representante de células eucarióticas saudáveis. Em particular, dados de espectroscopia de absorção de reflexão de infravermelho modulada por polarização (PM-IRRAS) revelaram um aumento da ordem conformacional das caudas de DOPS devido à adsorção mais forte de AuSHINs. Além disso, a irradiação reduziu a estabilidade de AuSHINs contendo monocamadas de DOPC e DOPS devido às reações oxidativas desencadeadas por ROS mediante aquecimento fotoinduzido. Em comparação com o DOPC, o DOPS perdeu quase o dobro de material para a subfase, o que é consistente com uma taxa mais alta de formação de ROS na vizinhança da monocamada DOPS. Metallic nanoparticles are promising agents for photothermal cancer therapy (PTT) owing to their photostability and efficient light-to-heat conversion, but their possible aggregation remains an issue. In this paper, we report on the photoinduced heating of gold shell-isolated nanoparticles (AuSHINs) in in vitro experiments to kill human oropharyngeal (HEp-2) and breast (BT-474 and MCF-7) carcinoma cells, with cell viability reducing below 50 % with 2.2 × 1012 AuSHINs/mL and 6 h of incubation. This toxicity to cancer cells is significantly higher than in previous works with gold nanoparticles. Considering the AuSHINs dimensions we hypothesize that cell uptake is not straightforward, and the mechanism of action involves accumulation on phospholipid membranes as the PTT target for photoinduced heating and subsequent generation of reactive oxygen species (ROS). Using Langmuir monolayers as simplified membrane models, we confirmed that AuSHINs have a larger effect on 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (DOPS), believed to represent cancer cell membranes, than on 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) taken as representative of healthy eukaryotic cells. In particular, data from polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) revealed an increased conformational order of DOPS tails due to the stronger adsorption of AuSHINs. Furthermore, light irradiation reduced the stability of AuSHINs containing DOPC and DOPS monolayers owing to oxidative reactions triggered by ROS upon photoinduced heating. Compared to DOPC, DOPS lost nearly twice as much material to the subphase, which is consistent with a higher rate of ROS formation in the vicinity of the DOPS monolayer. Postprint Universidade Estadual Paulista (UNESP), Faculdade de Ciências e Letras, Assis Universidade de São Paulo (USP), Instituto de Física de São Carlos (IFSC) FAPESP: 2018/14692-5 FAPESP: 2018/16713-0 FAPESP: 2013/14262-7 FAPESP: 2018/22214- 6 CNPq: 403713/2016-1

Made available in DSpace on 2020-12-12T02:41:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-08-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Instituto Nacional de Ciência e Tecnologia em Eletrônica Orgânica Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Risks to human health have been reported owing to prolonged exposure to hormones, whose action depends on their molecular-level interaction with cell membranes. In this study, we investigate the interaction of the synthetic hormone 17 α-ethynylestradiol (EE2) in two different membrane models, Langmuir monolayers and giant unilamellar vesicles (GUVs) made with a binary mixture of 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) and cholesterol (Chol) in order to mimic the plasma membrane of mammalian cells. EE2 was found to expand the Langmuir monolayers, with shifts to larger areas per molecule in the surface pressure isotherm. In all of these observations, stronger effects were noted for the DPPC/Chol monolayers with XChol = 0.3, which mimics the proportion of phospholipid/sterol in the plasma membrane. At high surface pressures, EE2 is believed to weaken the attractive interactions between DPPC and Chol, in addition to affecting the ordering of the lipid chains as indicated in polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) measurements. In GUVs obtained with XChol = 0.3 mixtures, EE2 induced a phase contrast loss as a result of increased permeability of the lipid bilayer. The results with Langmuir monolayers and GUV combined point to EE2 action on representative cell membranes, which can be correlated with physiological effects caused by indirect intake of EE2. Department of Physics School of Technology and Applied Sciences São Paulo State University (UNESP) Sao Carlos Institute of Physics University of Sao Paulo (USP) Department of Physics School of Technology and Applied Sciences São Paulo State University (UNESP) FAPESP: 2013/14262-7 FAPESP: 2016/09633-4 CAPES: 88882.330129/2018-01

Bone biomineralization is an exquisite process by which a hierarchically organized mineral matrix is formed. Growing evidence has uncovered the involvement of one class of extracellular vesicles, named matrix vesicles (MVs), in the formation and delivery of the first mineral nuclei to direct collagen mineralization. MVs are released by mineralization-competent cells equipped with a specific biochemical machinery to initiate mineral formation. However, little is known about the mechanisms by which MVs can trigger this process. Here, we present a combination of in situ investigations and ex vivo analysis of MVs extracted from growing-femurs of chicken embryos to investigate the role played by phosphatidylserine (PS) in the formation of mineral nuclei. By using self-assembled Langmuir monolayers, we reconstructed the nucleation core - a PS-enriched motif thought to trigger mineral formation in the lumen of MVs. In situ infrared spectroscopy of Langmuir monolayers and ex situ analysis by transmission electron microscopy evidenced that mineralization was achieved in supersaturated solutions only when PS was present. PS nucleated amorphous calcium phosphate that converted into biomimetic apatite. By using monolayers containing lipids extracted from native MVs, mineral formation was also evidenced in a manner that resembles the artificial PS-enriched monolayers. PS-enrichment in lipid monolayers creates nanodomains for local increase of supersaturation, leading to the nucleation of ACP at the interface through a multistep process. We posited that PS-mediated nucleation could be a predominant mechanism to produce the very first mineral nuclei during MV-driven bone/cartilage biomineralization.

Gram-negative bacteria possess numerous defenses against antibiotics, due to the intrinsic permeability barrier of their outer membrane (OM), explaining the recalcitrance of some common and life-threatening infections. We report the formulation of a new drug, PPA148, which shows promising activity against all Gram-negative bacteria included in the ESKAPEE pathogens. PPA148 was solubilized by inclusion complexation with cyclodextrin followed by encapsulation in liposomes. The complex and liposomal formulation presented increased activity against E. coli compared to the pure drug when assessed with the Kirby Bauer assay. The novel formulation containing 1 μg PPA148 reached similar efficacy levels equivalent to those of 30 μg of pure rifampicin. A range of biophysical techniques was used to explore the mechanism of drug uptake. Langmuir trough (LT) and neutron reflectivity (NR) techniques were employed to monitor the interactions between the drug and the formulation with model membranes. We found evidence for liposome fusion with the model Gram-negative outer membrane and for cyclodextrins acting as inner membrane (IM) permeation enhancers without presenting intrinsic antimicrobial activity. An antibiotic-in-cyclodextrin-in-liposomes (ACL) formulation was developed, which targets both the bacterial OM and IM, and offers promise as a means to breach the Gram-negative cell envelope.

(1) Background: The study characterized barley mutants with brassinosteroid (BR) biosynthesis and signaling disturbances in terms of the physicochemical/structural properties of membranes to enrich the knowledge about the role of brassinosteroids for lipid metabolism and membrane functioning. (2) Methods: The Langmuir method was used to investigate the properties of the physicochemical membranes. Langmuir monolayers were formed from the lipid fractions isolated from the plants growing at 20 &deg, C and then acclimated at 5 &deg, C or 27 &deg, C. The fatty acid composition of the lipids was estimated using gas chromatography. (3) Results: The BR-biosynthesis and BR-signaling mutants of barley were characterized by a temperature-dependent altered molar percentage of fatty acids (from 14:0 to 20:1) in their galactolipid and phospholipid fractions in comparison to wild-type (WT). For example, the mutants had a lower molar percentage of 18:3 in the phospholipid (PL) fraction. The same regularity was observed at 5 &deg, C. It resulted in altered physicochemical parameters of the membranes (Alim, &pi, coll, Cs&minus, 1). (4) Conclusions: BR may be involved in regulating fatty acid biosynthesis or their transport/incorporation into the cell membranes. Mutants had altered physicochemical parameters of their membranes, compared to the WT, which suggests that BR may have a multidirectional impact on the membrane-dependent physiological processes.

The influence of four common cryoprotectants (dimethyl sulfoxide, glycerol, ethylene glycol and dimethylformamide) on monolayers of four common phospholipids (DPPC, DOPC, POPC and POPE) have been studied using Langmuir isotherms and monolayer insertion experiments. The cryoprotectant concentrations were chosen to be directly relevant to cryoprotection. We show that DMSO causes an expansion of the DPPC area per lipid (in contrast to previous work at higher concentrations). However, it caused compression for POPC, and had little effect for POPE or DOPC. As most previous studies have involved only DPPC, this highlights the importance of studying different lipid types as these may have a significant effect on the interactions. We show that both ethylene glycol and glycerol cause a small expansion of the monolayer at fixed pressure, implying that they insert into the headgroup regions, regardless of lipid species, and consistent with their ability to penetrate membranes. By contrast, dimethylformamide causes monolayer compression for all lipid species, implying it dehydrates the lipid head groups. Membrane insertion experiments at physiological values of lateral pressure highlight that DPPC is the most difficult lipid to penetrate, implying that the penetrating action of cryoprotectants may only occur for unsaturated phospholipids. Thus, extrapolations of results based solely on the DPPC need to be made with care

Soil bacteria are decomposer organisms crucial for the biodegradation of organic pollutants, mineralization of dead organic matter and the turnover of biogenic elements. In their environment they are constantly exposed to membrane-lytic enzymes emitted to the soil by other microorganisms competing for the same niche. Therefore, the composition and structure of their membranes is of utmost importance for survival in the harsh environment. Although soil bacteria species can be Gram-negative or Gram-positive and their membranes differ significantly, they are formed by phospholipids belonging mainly to three classes: phosphatidylethanolamines (PE), phosphatidylglycerols (PG) and cardiolipins (CL). The correlation of the membrane phospholipid composition and its susceptibility to secretory membrane-lytic enzymes is widely unknown; thus, to shed light on these phenomena we applied the Langmuir monolayer technique to construct models of soil bacteria membranes differing in the mutual proportion of the main phospholipids. To characterize the systems we studied their elasticity, mesoscopic texture, 2D crystalline structure and discussed the thermodynamics of the interactions between their components. The model membranes were exposed to secretory phospholipase A2. It turned out that in spite of the structural similarities the model membranes differed significantly in their susceptibility to s-PLA2 attack. The membranes devoid of cardiolipin were completely degraded, whereas, these containing cardiolipin were much more resistant to the enzymatic hydrolysis. It also turned out that the sole presence of cardiolipin in the model membrane did not guarantee the membrane durability and that the interplay between cardiolipin and the zwitterionic phosphatidylethanolamine was here of crucial importance.

Lipid layers are considered among the first protective barriers of the human body against pollutants, e.g., skin, lung surfactant, or tear film. This makes it necessary to explore the physico-chemical bases underlying the interaction of pollutants and lipid layers. This work evaluates using a pool of surface-sensitive techniques, the impact of carbon black and fumed silica particles on the behavior of Langmuir monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The results show that the incorporation of particles into the lipid monolayers affects the surface pressure&ndash, area isotherm of the DPPC, modifying both the phase behavior and the collapse conditions. This is explained considering that particles occupy a part of the area available for lipid organization, which affects the lateral organization of the lipid molecules, and consequently the cohesion interactions within the monolayer. Furthermore, particles incorporation worsens the mechanical performance of lipid layers, which may impact negatively in different processes presenting biological relevance. The modification induced by the particles has been found to be dependent on their specific chemical nature. This work tries to shed light on some of the most fundamental physico-chemical bases governing the interaction of pollutants with lipid layers, which plays an essential role on the design of strategies for preventing the potential health hazards associated with pollution.

Local anesthetics (LAs) are known to act on membrane level; however, the molecular mechanism of their activity is still not fully understood. One hypothesis holds that these drugs can incorporate into lipid membrane of nerve cells and in this way change conformation of channel proteins responsible for transport of sodium ions. However, the action of anesthetics is not limited to nerve cells. These drugs also affect other types of cells and organelles, causing severe side effects. In this paper, we applied Langmuir monolayers—as model of cellular membranes—and investigated interactions between selected amide-type local anesthetics (lidocaine prilocaine, mepivacaine and ropivacaine, in the form of hydrochlorides) and lipid components of natural membranes: cholesterol, POPC and cardiolipin (CL) and their mixtures (POPC/cholesterol and POPC/CL/cholesterol), which can serve as simplified models of nerve cell membranes, erythrocytes, and mitochondria. The influence of the drug was monitored by registering the surface pressure (π) as a function of surface area per molecule (A) in a monolayer in the presence of the drug in the subphase. The structure of lipid monolayers on subphases containing and devoid of the studied drugs were visualized with Brewster angle microscopy (BAM). Langmuir monolayer studies complemented with surface visualization technique reveal the expansion and fluidization of lipid monolayers, with the most pronounced effect observed for cardiolipin. In mixed systems, the effect of LAs was found to depend on cholesterol proportion. The observed fluidization of membranes by local anesthetics may negatively affect cells functioning and therefore can explain side effects of these drugs both on the cardiovascular and nervous systems. Electronic supplementary material The online version of this article (10.1007/s00232-018-0055-6) contains supplementary material, which is available to authorized users.

The progressive contamination of food products by mycotoxins such as zearalenone (ZEN) has prompted the search for specific substances that can act as protectors against an accumulation of these toxins. This paper discusses the effect of selenium ions and 24-epibrassinolide (EBR) as non-organic and organic compounds that preserve human lymphoblastic cells U-937 under ZEN stressogenic conditions. Based on measurements of cell viability and a DAPI test, concentrations of ZEN at 30 μmol/l, Se at 2.5 μmol/l and EBR at 0.005 μmol/l were selected. The addition of both protectors resulted in an increase in the viability of ZEN-treated cells by about 16%. This effect was connected with a decrease in lipid peroxidation (a decrease in the malonyldialdehyde content) and the generation of reactive oxygen species, which were determined by a cellular ROS/superoxide detection assay and the SOD activity. The Se protection was observed as the blocking of the all excess ROS, while the EBR action was mainly concentrated on something other than the superoxide radical itself. The experiments on the model lipid membranes that mimic the environment of U-937 cells confirmed the affect of ZEN on the structure and physicochemical properties of human membranes. Although the presence of both Se and EBR reduced the effect of ZEN by blocking its interaction with a membrane, the action of Se was more evident.

PbS nanocrystals are an important narrow-gap material for solar cells and photodetectors. Nevertheless, their application may be limited because device performance can be affected by atmospheric conditions. Indeed, the presence of oxygen and/or water can degrade the active layers, possibly leading to device failure. Strategies to address this issue are therefore actively explored. Here we report a solution-processed PbS quantum dot solar cell, consisting of a PbS-silane functionalized reduced graphene oxide (PbS-rGO) layer on top of the PbS absorber film, which enhances device stability, especially when the solar cells are exposed to moisture. Power conversion efficiency (PCE) measurements demonstrate a slower degradation under continuous illumination for solar cells with PbS-rGO. When storing the samples under saturated water vapor, differences are even more remarkable: with PbS-rGO the solar cells essentially maintain their initial PCE, while the PCE of the PbS reference devices is reduced by 50% after 5 days. Scanning electron microscopy, energy dispersive X-ray and X-ray photoelectron spectroscopy reveal the damage to the PbS films and the formation of PbSOx crystals in the PbS reference devices. Such crystals are not observed in the PbS-rGO devices, further supporting the importance of the PbS-rGO barrier layer.

Glycosaminoglycans (GAGs) are essential components of the extracellular matrices (ECMs) located on the outer surface of cellular membranes. They belong to the group of polysaccharides involved in diverse biological processes acting on the surface and across natural lipid membranes. Recently, particular attention has been focused on possible role of GAGs in the amyloid deposits. The amyloid formation is related to a disorder in protein folding, causing that soluble—in normal conditions—peptides become deposited extracellularly as insoluble fibrils, impairing tissue structure and its function. One of the hypothesis holds that GAGs may inhibit amyloid formation by interacting with the lipid membrane by blocking the accumulation of protein aggregates on the membrane surface. Although the biophysical properties of GAGs are described rather well, little is known about the nature of association between these polysaccharides and components of natural cell membranes. Therefore, a study of GAGs influence on membrane lipids is of particular importance. The aim of the present work is to get insight into the effect of hydrophilic dextran sulfate (DS)—that can be considered as GAG analogue—on membrane lipids organization. This study was based on examining interactions between DS sodium salt of molecular weight equal to about 40 kDa (DS40), dissolved in water subphase, and a model membrane, mimicked as Langmuir monolayer, formed by representative natural membrane lipids: cholesterol and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) as well as their mixtures. Due to the fact that calcium ions in excess may accumulate in the lipid membrane, attracting high molecular weight molecules to their surface, the influence of calcium ions present in the subphase on the DS40 activity has also been examined. It has been found that negatively charged DS, forming a sublayer underneath the monolayer, barely interacts with membrane lipids; however, in the presence of calcium ions the electrostatic interactions between DS40 and lipid membrane are significantly enhanced, leading to the formation of network-like crystalline structures at the surface of model membrane, which can prevent incorporation and interaction with other extracellular molecules, e.g., proteins.

Measuring surface (excess) entropies provides a bounty of valuable structural information that is hard to obtain otherwise. In the paper these quantities are defined and procedures of measurements discussed. Mostly they involve measurements at different temperatures. A review is given for interfaces with aqueous solutions in the absence of polymers. This review illustrates how, sometimes unanticipated, pieces of information are obtained, for example with cloud seeding and a possible explanation of the Jones-Ray effect. As a novel extension the procedure is applied to deposited, or Langmuir, monolayers of poly(ethylene oxide)-poly(propylene oxide) block copolymers. It will be shown how the various phase transitions and associated configurations of these polymers can be recognized and monitored.

Organic macromolecules with dendrimeric architectures are polymeric materials potentially useful as nanocarriers for therapeutic drugs. In this work, we evaluate a series of Newkome-type dendrons in Langmuir and Langmuir-Blodgett films as platforms capable of interacting with a potential antitumoral agent. The nanocomposite is proposed as model for the development of surface mediated drug delivery systems. We were successful in the formation and characterization of pure (dendrons) and composite (drug-dendron) stable and reproducible monolayers, and their transfer to solid substrates. A detailed study of topographic characteristics of the generated surfaces by atomic force microscopy was conducted. Furthermore, we probed dendron monolayer films as anchorage surfaces for mammalian cells. Normal cell attachment and proliferation on the surfaces were observed. No evident cytotoxic effects were detected, demonstrating the adequate biocompatibility of the surfaces. Fil: Dib, Nahir. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Reviglio, Ana Lucía. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Fernández, Luciana. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Morales, Gustavo. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Santo, Marisa Rosana. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Otero, Luis Alberto. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química y Física; Argentina Fil: Alustiza, Fabrisio Eduardo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina Fil: Liaudat, Ana Cecilia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina Fil: Bosch, Pablo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina Fil: Calderon, Marcelo. Universität zu Berlin; Alemania. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Departamento de Química Orgánica; Argentina Fil: Martinelli, Marisa. Universidad Nacional de Córdoba. Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada; Argentina Fil: Strumia, Miriam Cristina. Universidad Nacional de Córdoba. Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación y Desarrollo en Ingeniería de Procesos y Química Aplicada; Argentina

The dynamic surface properties of mixed monolayers of 1,2-Dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) and negatively charged polystyrene particles (PS) of different size were investigated with the aim to model the interactions of particles with biointerfaces. Special attention was paid to the aggregation of the particles at the water/air interface. The particle aggregates have a weaker influence on the dynamic properties of DPPC monolayers as compared with non-aggregated particles because of a shorter three phase contact line. The incorporation of particles into a DPPC monolayer leads to disordering of the monolayer structure, decreases the changes of surface properties in the course of two-dimensional phase transitions and the dynamic surface elasticity of the condensed lipid film. The influence of particles on the properties of mixed monolayers depends strongly on the ratio of lipid molecules and particles, and decreases with the decrease of particle surface concentration. The Brewster angle microscopy gives direct evidence of the PS particle aggregation in the course of compression of mixed monolayers.

Mono- and digalactosyldiacylglycerol are essential galactolipids for the biogenesis of plastids and functioning of the photosynthetic machinery. In Arabidopsis, the first step of galactolipid synthesis is catalyzed by monogalactosyldiacylglycerol synthase 1 (MGD1), a monotopic protein located in the inner envelope membrane of chloroplasts, which transfers a galactose residue from UDP-galactose to diacylglycerol (DAG). MGD1 needs anionic lipids such as phosphatidylglycerol (PG) to be active, but the mechanism by which PG activates MGD1 is still unknown. Recent studies shed light on the catalytic mechanism of MGD1 and on the possible PG binding site. Particularly, Pro189 was identified as a potential residue implicated in PG binding and His155 as the putative catalytic residue. In the present study, using a multifaceted approach (Langmuir membrane models, atomic force microscopy, molecular dynamics; MD), we investigated the membrane binding properties of native MGD1 and mutants (P189A and H115A). We demonstrated that both residues are involved in PG binding, thus suggesting the existence of a PG-His catalytic dyad that should facilitate deprotonation of the nucleophile hydroxyl group of DAG acceptor. Interestingly, MD simulations showed that MGD1 induces a reorganization of lipids by attracting DAG molecules to create an optimal platform for binding.

The interaction of hydrophobic silver nanoparticles with different phospholipids and stratum corneum mimic (SCM) membranes is studied in Langmuir monolayers. Thus, silver nanoparticles coated with oleic acid (AgNP-OA) were synthesized, characterized and incorporated in Langmuir monolayers of single phospholipids –having different chain length, saturation degree and phase state– or of a SCM mixture. The incorporation of AgNP-OA to the lipid monolayers generated an expansion of the monolayers and a decrease of the surface compressional modulus compared to the pure lipid. X-ray photoelectron spectroscopy (XPS) suggested that the zwitterionic choline-phospholipids can be adsorbed onto the nanoparticles’ surface, which is relevant considering that phospholipids are the major constituents of the cell membrane. We also studied the changes in the topography at the mesoscale level using Brewster angle microscopy. We found the most prominent changes in the lipids with liquid-condensed phase, such as SCM, showing segregation of their components. This could have major implications in the barrier function of the membrane, affecting for example the skin permeability towards hydrophobic nanoparticles. Finally, the capability of hydrophobic AgNP-OA for delivering Ag + ions was studied in aqueous media in the absence and presence of phospholipids. In both conditions, AgNP-OA released Ag + at reported-bactericidal concentrations, being double in the presence of phospholipids. Fil: Villanueva, Martín Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Lanterna, Anabel Estela. University of Ottawa; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Vico, Raquel Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

33 p.-6 fig., Phasins are amphiphilic proteins located at the polymer-cytoplasm interface of bacterial polyhydroxyalkanoates (PHA). The immobilization of phasins on biomaterial surfaces is a promising way to enhance the hydrophilicity and supply cell-directing elements in bioinstructing processes. Optimizing the physical adsorption of phasins requires deep insights into molecular processes during polymer-protein interactions to preserve their structural conformation while optimizing surface coverage. Here, the assembly, organization and stability of phasin PhaF from Pseudomonas putida at interfaces is disclosed. The Langmuir technique, combined with in situ microscopy and spectroscopic methods, revealed that PhaF forms stable and robust monolayers at different temperatures, with an almost flat orientation of its alpha-helix at the air-water interface. PhaF adsorption to preformed monolayers of poly[(3-R-hydroxyoctanoate)-co-(3-R-hydroxyhexanoate)] (PHOHHx), yields stable mixed-layers below π = ~15.7 mN/m. Further insertion induces a molecular reorganization. PHOHHx with strong surface hydrophobicity is a more adequate substrate for PhaF adsorption than the less hydrophobic poly[(rac-lactide)-co-glycolide] (PLGA). The observed orientation of the main axis of the protein in relation to copolyester interfaces ensures the best exposure of the hydrophobic residues, providing a suitable coating strategy for functionalized polymers., The work was funded by the Helmholtz Graduate School Macromolecular Bioscience (no. VH-GS-503) and the Helmholtz Programme-oriented Funding. Additional funding was provided by the European Union’s Horizon 2020 research and innovation program (nº 633962), Ministerio de Ciencia Innovación y Universidades (BIO2017-83448-R), and the PhD Fellowship program of the Department of Science Technology and Innovation-Colciencias, Colombia. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

New gemini cationic surfactants bearing imidazolium groups as polar heads and alkyl chains of various lengths were prepared using the amino acid histidine as starting material. These surfactants were found to have very low cmc values and reduced the surface tension of water solutions with high efficiency. Their biological properties, antimicrobial and hemolytic activities, depended on the alkyl chain length. These surfactants are active against a wide range of bacteria including methicillin-resistant Staphylococcus aureus (MRSA). Moreover, these gemini surfactants have good DNA binding capacity. These new gemini surfactants show a great potential in cosmetic, food and pharmaceutical industries., This work was financed by the Ministerio de Economía y Competitividad, Spain, grants CTQ2017-88948-P and CTQ2014-59632-R. Authors also thank AGAUR 2017 SGR 670.

Hypothesis Amphotericin B (AmB) is a highly effective antimicrobial, with broad antimycotic and antiparasitic effect. However, AmB poor water-solubilisation and aggregation tendency limits its use for topical applications. We studied the capacity of nanostructures formed by alkyl esters of L-ascorbic acid (ASCn) to solubilise AmB and tested the relationship between the prevalence of the monomeric form of AmB and its effectiveness as antimicrobial agent. Experiments We developed self-assembled nanostructures formed by the commercial compound, palmitoyl ascorbic acid, as well as the shorter chained myristoyl and lauroyl ascorbic acid. AmB loaded ASCn nanostructures were studied by a combination of spectroscopic techniques, together with particle analysis, differential scanning calorimetry, microbiological tests, and Langmuir monolayer visualisation. Findings We found no direct relation between the antimicrobial capacity and the prevalence of the monomeric form of the drug. However, the later was related to chemical stability and colloidal robustness. Nanostructures formed by ASC16 in its anionic state provide an appropriate environment for AmB in its monomeric form, maintaining its antimicrobial capacity. Langmuir film visualisation supports spectrophotometric evidence, indicating that ASC16 allows the in-plane solubilisation of AmB. Coagels formed by ASC16 appear as promising for carrying AmB for dermal delivery., amphiphilic drugs; Langmuir monolayers; coagels; polyene macrolides; drug carriers.

The molecular packing, thermodynamics and surface topography of binary Langmuir monolayers of Insulin and DPPC (dipalmitoylphosphatidylcholine) or POCP (1-palmitoyl-2-oleoylphosphatidylcholine) at the air/water interface on Zn2+ containing solutions were studied. Miscibility and interactions were ascertained by the variation of surface pressure-mean molecular area isotherms, surface compressional modulus and surface (dipole) potential with the film composition. Brewster Angle Microscopy was used to visualize the surface topography of the monolayers. Below 20mN/m Insulin forms stable homogenous films with DPPC and POPC at all mole fractions studied (except for films with XINS=0.05 at 10mN/m where domain coexistence was observed). Above 20mN/m, a segregation process between mixed phases occurred in all monolayers without squeezing out of individual components. Under compression the films exhibit formation of a viscoelastic or kinetically trapped organization leading to considerable composition-dependent hysteresis under expansion that occurs with entropic-enthalpic compensation. The spontaneously unfavorable interactions of Insulin with DPPC are driven by favorable enthalpy that is overcome by unfavorable entropic ordering; in films with POPC both the enthalpic and entropic effects are unfavorable. The surface topography reveals domain coexistence at relatively high pressure showing a striped appearance. The interactions of Insulin with two major membrane phospholipids induces composition-dependent and long-range changes of the surface organization that ought to be considered in the context of the information-transducing capabilities of the hormone for cell functioning. Fil: Grasso, Ernesto Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina Fil: Oliveira, Rafael Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina Fil: Maggio, Bruno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina

The incorporation of silica nanoparticles (NPs) from the subphase into Langmuir lipid monolayers formed by three components, 1,2-Dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and Cholesterol (Chol), modifies the thermodynamic and rheological behavior, as well as the structure of the pristine lipid film. Thus, the combination of structural characterization techniques, such as Brewster Angle Microscopy (BAM) and Atomic Force Microscopy (AFM), with interfacial thermodynamic and dilational rheology studies has allowed us to deepen on the physico-chemical bases governing the interaction between lipid molecules and NPs. The penetration of NPs driven by the interaction (electrostatic or hydrogen bonds) with the polar groups of the lipid molecules affects the phase behaviour (surface pressure-area, ? - A, isotherm) of the monolayer. This can be easily rationalized considering the modification of the packing and cohesion of the molecules at the interface as revealed BAM and AFM images. Furthermore, oscillatory barrier experiments have allowed obtaining information related to the effect of NPs on the monolayer response under dynamic conditions that presents a critical impact on the characterization of biological relevant systems because most of the processes of interest for these systems present a dynamic character.

Cholesterol (Chol) interacts with lipoproteins, in order to be transported through the aqueous bloodstream. High density lipoproteins (HDL) and low density lipoproteins (LDL) transport cholesterol differently, a result that may be due to a difference in their interactions with cholesterol. Here, we investigated how the lipoprotein type affects the interaction with cholesterol by using a Langmuir trough and fluorescence microscope. We studied pure monolayers of 1) Chol, 2) LDL, and 3) HDL, and mixed monolayers of 1) Chol-LDL, and 2) Chol-HDL at air/water interfaces. Images of the Chol-LDL mixed monolayer showed many small sterol domains distributed in the non-sterol molecules (e.g. phospholids, proteins and lipids) of LDL. The sterol domains that were seen in the Chol-HDL mixed monolayer were larger in size but smaller in number than those seen in the Chol-LDL mixed monolayers. These images and the excess area, excess free energy, and free energy of mixing values obtained from the thermodynamic analysis of the surface pressure-area per molecule isotherms suggested that the cholesterol phase separated more from HDL than from LDL. Cholesterol was therefore concluded to interact with LDL better than with HDL. This more favorable interaction was explained by the presence of hydrophobic interactions between cholesterol and Apo-B, the major apoprotein of LDL., Bioengineering, Biophysics, Physical chemistry, Thermodynamics, High density lipoproteins; Low density lipoproteins; Cholesterol; Langmuir monolayers; Fluorescence imaging

The use of reversible addition-fragmentation chain transfer (RAFT)-assisted encapsulating emulsion polymerization (REEP) has been explored to prepare diverse types of colloidal stable core&ndash, shell nanostructures. A major field of application of such nanoparticles is in emergent nanomedicines, which require effective biofunctionalization strategies, in which their response to bioanalytes needs to be firstly assessed. Herein, functional core&ndash, shell nanostructures were prepared via REEP and click chemistry. Thus, following the REEP strategy, colloidal gold nanoparticles (Au NPs, d = 15 nm) were coated with a poly(ethylene glycol) methyl ether acrylate (PEGA) macroRAFT agent containing an azide (N3) group to afford N3&ndash, macroRAFT@Au NPs. Then, chain extension was carried out from the NPs surface via REEP, at 44 &deg, C under monomer-starved conditions, to yield N3&ndash, copolymer@Au NPs&ndash, core&ndash, shell type structures. Biotin was anchored to N3-copolymer@Au NPs via click chemistry using an alkynated biotin to yield biofunctionalized Au nanostructures. The response of the ensuing biotin&ndash, copolymer@Au NPs to avidin was followed by visible spectroscopy, and the copolymer&ndash, biotin&ndash, avidin interaction was further studied using the Langmuir&ndash, Blodgett technique. This research demonstrates that REEP is a promising strategy to prepare robust functional core&ndash, shell plasmonic nanostructures for bioapplications. Although the presence of azide moieties requires the use of low polymerization temperature, the overall strategy allows the preparation of tailor-made plasmonic nanostructures for applications of biosensors based on responsive polymer shells, such as pH, temperature, and photoluminescence quenching. Moreover, the interaction of biotin with avidin proved to be time dependent.

Cell membrane models have been used to evaluate the interactions of various imidazolium-based ionic liquids (ILs) with Langmuir monolayers of two types of phospholipids and cholesterol. Data from surface pressure isotherms, Brewster angle microscopy (BAM) and polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) pointed to significant effects on the monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol, used to mimic the membranes of eukaryotic cells, for ILs containing more than 6 carbon atoms in the alkyl chain (i.e. n > 6). For ILs with less hydrophobic tails (n ≤ 6) and low concentrations, the effects were almost negligible, therefore, such ILs should not be toxic to eukaryotic cells. The hydrophobicity of the anion was also proved to be relevant, with larger impact from ILs containing tetrafluoroborate ([BF4]-) than chloride (Cl-). Molecular dynamics simulations for DPPC monolayers at the surface of aqueous solutions of alkylimidazolium chloride ([Cnmim]Cl) confirm the penetration of the IL cations with longer alkyl chains into the phospholid monolayer and provide information on their location and orientation within the monolayer. For monolayers of dipalmitoylphosphatidyl glycerol (DPPG), which is negatively charged like bacteria cell membranes, the ILs induced much larger effects. Similarly to the results for DPPC and cholesterol, effects increased with the number of carbon atoms in the alkyl chain and with a more hydrophobic anion [BF4]-. Overall, the approach used can provide relevant information of molecular-level interactions behind the toxicity mechanisms and support the design of (quantitative) structure-activity relationship models, which may help design more efficient and environmentally friendly ILs. published

Membrane structure is a key factor for the cell`s physiology, pathology, and therapy. Evaluating the importance of lipid species such as N-nervonoyl sphingomyelin (24:1-SM) -able to prevent phase separation- to membrane structuring remains a formidable challenge. This is the first report in which polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS) is applied to investigate the lipid-lipid interactions in 16:0 vs 24:1-SM monolayers and their mixtures with 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol (Chol) (DOPC/SM/Chol 2:1:1). From the results we inferred that the cis double bond (Δ15) in 24:1-SM molecule diminishes intermolecular H-bonding and chain packing density compared to that of 16:0-SM. In ternary mixtures containing 16:0-SM, the relative intensity of the two components of the Amide I band reflected changes in the H-bonding network due to SM-Chol interactions. In contrast, the contribution of the main components of the Amide I band in DOPC/24:1-SM/Chol remained as in 24:1-SM monolayers, with a larger contribution of the non-H-bonded component. The most interesting feature in these ternary films is that the CO stretching mode of DOPC appeared with an intensity similar to that of SM Amide I band in DOPC/16:0-SM/Chol monolayers (a two-phase [Lo/Le] system), whereas an extremely low intensity of the CO band was detected in DOPC/24:1-SM/Chol monolayers (single Le phase). This is evidence that the unsaturation in 24:1-SM affected not only the conformational properties of acyl chains but also the orientation of the chemical groups at the air/water interface. The physical properties and overall H-bonding ability conferred by 24:1-SM may have implications in cell signaling and binding of biomolecules., Instituto de Investigaciones Bioquímicas de La Plata, Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas

Made available in DSpace on 2018-12-11T16:54:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Photodynamic therapy (PDT) efficiency depends on many factors including the incorporation of the photosensitizer (PS) in cell membranes and possible lipid hydroperoxidation. In this study, we show that hydroperoxidation may be photoinduced when eosin Y is incorporated into Langmuir monolayers that serve as cell membrane models. This occurs for Langmuir monolayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), which have unsaturation in their hydrophobic chains. In contrast, light irradiation had no effect on monolayers of saturated 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Evidence of hydroperoxidation was obtained from the area increase in eosin-containing DOPC and POPC monolayers upon irradiation, which was accompanied by a decrease in monolayer thickness according to grazing incidence X-ray off-specular scattering (GIXOS) data. Furthermore, the changes in polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) induced by irradiation were consistent with hydroperoxide migration toward the lipid hydrophilic heads. In summary, this combination of experimental methods allowed us to determine the effects of eosin Y interaction with cell membrane models under irradiation, which may be associated with the underlying mechanisms of eosin Y as photosensitizer in PDT. School of Sciences Humanities and Languages São Paulo State University (UNESP) Brazilian Synchrotron Light Laboratory-LNLS CNPEM, Rua Giuseppe Maximo Scolfaro, 10000 IFSC São Carlos Institute of Physics University of São Paulo (USP) School of Sciences Humanities and Languages São Paulo State University (UNESP) FAPESP: 2013/14262-7 FAPESP: 2016/13280-0 FAPESP: 2017/13404-3 CNPq: 403713/2016-1

The interaction of carbon black particles (CB) with 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) at the water/air interface has been investigated by means of a pool of surface sensitive techniques, in order to analyze the thermodynamic and rheological aspects of these mixed systems. The incorporation of carbonaceous particles to the lipid monolayers induces changes on the surface pressure-area isotherm, as revealed by the shifting to higher surface area of the maximum packing degree of the monolayer, and the decrease of the collapse pressure. These changes are strongly dependent on the DPPC:CB weight ratio at the interface and can be explained by the disruption of the monolayer structure due to the particle incorporation that provokes the modification of the cohesive interactions along the monolayer. Measurements of dilational viscoelasticity against frequency, at different degrees of monolayer compression, have been performed by means of the Oscillatory Barrier method. The rheological response of the monolayer is only slightly affected by the presence of CB, even if a modification of the quasi-equilibrium dilational elasticity, as well as of the frequency dependence of the viscoelastic modulus, is appreciable increasing the particle concentration. Being DPPC the major component of many systems with biological interest (cell membranes, lung surfactant), the results obtained here are expected to contribute to the understanding of the carbon particle interaction with biological relevant systems.

Outer layer of cellular membrane contains ordered domains enriched in cholesterol and sphingolipids, called 'lipid rafts', which play various biological roles, i.e., are involved in the induction of cell death by apoptosis. Recent studies have shown that these domains may constitute binding sites for selected drugs. For example alkylphosphocholines (APCs), which are new-generation antitumor agents characterized by high selectivity and broad spectrum of activity, are known to have their molecular targets located at cellular membrane and their selective accumulation in tumor cells has been hypothesized to be linked with the alternation of biophysical properties of lipid rafts. To get a deeper insight into this issue, interactions between representative APC: erucylphosphocholine, and artificial lipid raft system, modeled as Langmuir monolayer (composed of cholesterol and sphingomyelin mixed in 1:2 proportion) were investigated. The Langmuir monolayer experiments, based on recording surface pressure-area isotherms, were complemented with Brewster angle microscopy results, which enabled direct visualization of the monolayers structure. In addition, the investigated monolayers were transferred onto solid supports and studied with AFM. The interactions between model raft system and erucylphosphocholine were analyzed qualitatively (with mean molecular area values) as well as quantitatively (with ΔG(exc) function). The obtained results indicate that erucylphosphocholine introduced to raft-mimicking model membrane causes fluidizing effect and weakens the interactions between cholesterol and sphingomyelin, which results in phase separation at high surface pressures. This leads to the redistribution of cholesterol molecules in model raft, which confirms the results observed in biological studies.