| 1. |
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| 2. |
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| 3. |
- Datta, A., et al.
(författare)
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Role of Aromatic Amino Acids in Lipopolysaccharide and Membrane Interactions of Antimicrobial Peptides for use in Plant Disease Control
- 2016
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Ingår i: Journal of Biological Chemistry. - 0021-9258 .- 1083-351X. ; 291:25, s. 13301-13317
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Tidskriftsartikel (refereegranskat)abstract
- KYE28(KYEITTIHNLFRKLTHRLFRRNFGYTLR), the representative sequence of helix D of heparin co-factor II, was demonstrated to be potent against agronomically important Gram-negative plant pathogens X. vesicatoria and X. oryzae,capable of inhibiting disease symptoms in detached tomato leaves. NMR studies in presence of lipopolysaccharide provided structural insights into the mechanisms underlying this, notably in relation to outer membrane permeabilisation. The three-dimensional solution structure of KYE28 in LPS is characterised by a N-ter helical segment, an intermediate loop and an extended C-ter. The two termini are in close proximity to each other via aromatic packing interactions, while the positively charged residues formed an exterior polar shell. To further demonstrate the importance of the aromatic residues for this, a mutant peptide KYE28A, with Ala substitutions at F11, F19, F23 and Y25 showed attenuated antimicrobial activity at high salt concentrations, as well as lower membrane disruption and LPS binding abilities compared to KYE28. In contrast to KYE28, KYE28A adopted an opened out helical structure in LPS with extended N- and C-ter and a small break in between the helical segments. Aromatic packing interactions were completely lost, although hydrophobic interaction between the side chains of hydrophobic residues were still partly retained, imparting an amphipathic character and explaining its residual antimicrobial activity and LPS binding as observed from ellipsometry and ITC. We thus present important structural aspects of KYE28, constituting an aromatic zipper, of potential importance, for the development of novel plant protection agents and therapeutic agents.
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| 4. |
- Lilja, AM, et al.
(författare)
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Neural Stem Cell Transplant-Induced Effect on Neurogenesis and Cognition in Alzheimer Tg2576 Mice Is Inhibited by Concomitant Treatment with Amyloid-Lowering or Cholinergic α7 Nicotinic Receptor Drugs
- 2015
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Ingår i: Neural plasticity. - : Hindawi Limited. - 1687-5443 .- 2090-5904. ; 2015, s. 370432-
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Tidskriftsartikel (refereegranskat)abstract
- Stimulating regeneration in the brain has the potential to rescue neuronal networks and counteract progressive pathological changes in Alzheimer’s disease (AD). This study investigated whether drugs with different mechanisms of action could enhance neurogenesis and improve cognition in mice receiving human neural stem cell (hNSC) transplants. Six- to nine-month-old AD Tg2576 mice were treated for five weeks with the amyloid-modulatory and neurotrophic drug (+)-phenserine or with the partialα7 nicotinic receptor (nAChR) agonist JN403, combined with bilateral intrahippocampal hNSC transplantation. We observed improved spatial memory in hNSC-transplanted non-drug-treated Tg2576 mice but not in those receiving drugs, and this was accompanied by an increased number of Doublecortin- (DCX-) positive cells in the dentate gyrus, a surrogate marker for newly generated neurons. Treatment with (+)-phenserine did however improve graft survival in the hippocampus. An accumulation ofα7 nAChR-expressing astrocytes was observed around the injection site, suggesting their involvement in repair and scarring processes. Interestingly, JN403 treatment decreased the number ofα7 nAChR-expressing astrocytes, correlating with a reduction in the number of DCX-positive cells in the dentate gyrus. We conclude that transplanting hNSCs enhances endogenous neurogenesis and prevents further cognitive deterioration in Tg2576 mice, while simultaneous treatments with (+)-phenserine or JN403 result in countertherapeutic effects.
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| 5. |
- Caselli, Lucrezia, et al.
(författare)
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Boosting Membrane Interactions and Antimicrobial Effects of Photocatalytic Titanium Dioxide Nanoparticles by Peptide Coating
- 2024
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Ingår i: Small. - : John Wiley and Sons Inc. - 1613-6810 .- 1613-6829.
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Tidskriftsartikel (refereegranskat)abstract
- Photocatalytic nanoparticles offer antimicrobial effects under illumination due to the formation of reactive oxygen species (ROS), capable of degrading bacterial membranes. ROS may, however, also degrade human cell membranes and trigger toxicity. Since antimicrobial peptides (AMPs) may display excellent selectivity between human cells and bacteria, these may offer opportunities to effectively “target” nanoparticles to bacterial membranes for increased selectivity. Investigating this, photocatalytic TiO2 nanoparticles (NPs) are coated with the AMP LL-37, and ROS generation is found by C11-BODIPY to be essentially unaffected after AMP coating. Furthermore, peptide-coated TiO2 NPs retain their positive ζ-potential also after 1–2 h of UV illumination, showing peptide degradation to be sufficiently limited to allow peptide-mediated targeting. In line with this, quartz crystal microbalance measurements show peptide coating to promote membrane binding of TiO2 NPs, particularly so for bacteria-like anionic and cholesterol-void membranes. As a result, membrane degradation during illumination is strongly promoted for such membranes, but not so for mammalian-like membranes. The mechanisms of these effects are elucidated by neutron reflectometry. Analogously, LL-37 coating promoted membrane rupture by TiO2 NPs for Gram-negative and Gram-positive bacteria, but not for human monocytes. These findings demonstrate that AMP coating may selectively boost the antimicrobial effects of photocatalytic NPs. © 2024 The Authors.
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| 6. |
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| 7. |
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| 8. |
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| 9. |
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| 10. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Interaction of Laponite with Membrane Components - Consequences for Bacterial Aggregation and Infection Confinement
- 2019
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Ingår i: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 11:17, s. 15389-15400
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Tidskriftsartikel (refereegranskat)abstract
- The antimicrobial effects of Laponite nanoparticles with or without loading of the antimicrobial peptide LL-37 was investigated along with their membrane interactions. The study combines data from ellipsometry, circular dichroism, fluorescence spectroscopy, particle size/ζ potential measurements, and confocal microscopy. As a result of the net negative charge of Laponite, loading of net positively charged LL-37 increases with increasing pH. The peptide was found to bind primarily to the outer surface of the Laponite nanoparticles in a predominantly helical conformation, leading to charge reversal. Despite their net positive charge, peptide-loaded Laponite nanoparticles did not kill Gram-negative Escherichia coli bacteria or disrupt anionic model liposomes. They did however cause bacteria flocculation, originating from the interaction of Laponite and bacterial lipopolysaccharide (LPS). Free LL-37, in contrast, is potently antimicrobial through membrane disruption but does not induce bacterial aggregation in the concentration range investigated. Through LL-37 loading of Laponite nanoparticles, the combined effects of bacterial flocculation and membrane lysis are observed. However, bacteria aggregation seems to be limited to Gram-negative bacteria as Laponite did not cause flocculation of Gram-positive Bacillus subtilis bacteria nor did it bind to lipoteichoic acid from bacterial envelopes. Taken together, the present investigation reports several novel phenomena by demonstrating that nanoparticle charge does not invariably control membrane destabilization and by identifying the ability of anionic Laponite nanoparticles to effectively flocculate Gram-negative bacteria through LPS binding. As demonstrated in cell experiments, such aggregation results in diminished LPS-induced cell activation, thus outlining a promising approach for confinement of infection and inflammation caused by such pathogens.
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| 11. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Nanoclay-induced bacterial flocculation for infection confinement
- 2020
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier. - 0021-9797 .- 1095-7103. ; 562, s. 71-80
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Tidskriftsartikel (refereegranskat)abstract
- Effects of size and charge of anionic nanoclays on their interactions with bacteria-mimicking lipid membranes, bacterial lipopolysaccharide (LPS), and Gram-negative bacteria were investigated using ellipsometry, dynamic light scattering, ζ-potential measurements, and confocal microscopy combined with Live/Dead staining. Based on particle size and charge density, three different anionic hectorite nanoclays were employed, and investigated in the presence and absence of the net cationic human antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES). In the absence of this peptide, the nanoclays were found not to bind to similarly anionic bacteria-mimicking model phospholipid membranes, nor to destabilize these. Similarly, while all nanoclays induced aggregation of Escherichia coli bacteria, the flocculated bacteria remained alive after aggregation. In contrast, LL-37 alone, i.e. in the absence of nanoclay particles, displays antimicrobial properties through membrane lysis, but does not cause bacterial aggregation in the concentration range investigated. After loading the nanoclays with LL-37, potent bacterial aggregation combined with bacterial membrane lysis was observed for all nanoclay sizes and charge densities. Demonstrating the potential of these combined systems for confinement of infection, LPS-induced NF-κB activation in human monocytes was found to be strongly suppressed after nanoclay-mediated aggregation, with a wide tolerance for nanoparticle size and charge density.
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| 12. |
- Saravanan, Rathi, et al.
(författare)
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Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides
- 2018
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Ingår i: Nature Communications. - : NATURE PUBLISHING GROUP. - 2041-1723. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.
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| 13. |
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| 14. |
- Siegel, G., et al.
(författare)
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Lipoprotein binding to anionic biopolyelectrolytes and the effect of glucose on nanoplaque formation in arteriosclerosis and Alzheimer's disease
- 2016
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Ingår i: Advances in Colloid and Interface Science. - : Elsevier BV. - 0001-8686 .- 1873-3727. ; 232, s. 25-35
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Tidskriftsartikel (refereegranskat)abstract
- Arteriosclerosis with its clinical sequelae (cardiac infarction, stroke, peripheral arterial occlusive disease) and vascular/Alzheimer dementia not only result in far more than half of all deaths but also represent dramatic economic problems. The reason is, among others, that diabetes mellitus is an independent risk factor for both disorders, and the number of diabetics strongly increases worldwide. More than one-half of infants in the first 6 months of life have already small collections of macrophages and macrophages filled with lipid droplets in susceptible segments of the coronary arteries. On the other hand, the authors of the Bogalusa Heart Study found a strong increase in the prevalence of obesity in childhood that is paralleled by an increase in blood pressure, blood lipid concentration, and type 2 diabetes mellitus. Thus, there is a clear linkage between arteriosclerosis/Alzheimer's disease on the one hand and diabetes mellitus on the other hand. Furthermore, it has been demonstrated that distinct apoE isoforms on the blood lipids further both arteriosclerotic and Alzheimer nanoplaque formation and therefore impair flow-mediated vascular reactivity as well. Nanoplaque build-up seems to be the starting point for arteriosclerosis and Alzheimer's disease in their later full clinical manifestation. In earlier work, we could portray the anionic biopolyelectrolytes syndecan/perlecan as blood flow sensors and lipoprotein receptors in cell membrane and vascular matrix. We described extensively molecular composition, conformation, form and function of the macromolecule heparan sulfate proteoglycan (HS-PG). In two supplementary experimental settings (ellipsometry, myography), we utilized isolated HS-PG for in vitro nanoplaque investigations and isolated human coronary artery segments for in vivo tension measurements. With the ellipsometry-based approach, we were successful in establishing a direct connection on a molecular level between diabetes mellitus on the one side and arteriosclerosis/Alzheimer's disease on the other side. Application of glucose at a concentration representative for diabetics and leading to glycation of proteins and lipids, entailed a significant increase in arteriosclerotic and Alzheimer nanoplaque formation. IDLapoE4/E4 was by far superior to IDLapoE3/E3 in plaque build-up, both in diabetic and non-diabetic patients. Recording vascular tension of flow-dependent reactivity in blood substitute solution and under application of different IDLapoE isoforms showed an impaired vasorelaxation for pooled IDL and IDLapoE4/E4, thus confirming the ellipsometric investigations. Incubation in IDLapoE0/E0 (apoE "knockout man"), however, resulted in a massive flow mediated contraction, also complemented" by strongly aggregated nanoplaques. In contrast, HDL was shown to present a powerful protection against nanoplaque formation on principle, both in the in vitro model and the in vivo scenario on the endothelial cell membrane. The competitive interplay with LDL is highlighted through the flow experiment, where flow-mediated, HDL-induced vasodilatation remains untouched by additional incubation with LDL. This is due to the four times higher affinity for the proteoglycan receptor of HDL as compared to LDL. Taken together, the studies demonstrate that while simplistic, the ellipsometry approach and the endothelial mimicking proteoglycan-modified surfaces provide information on the initial steps of lipoprotein-related plaque formation, which correlates with findings on endothelial cells and blood vessels, and afford insight into the role of lipoprotein deposition and exchange phenomena at the onset of these pathophysiologies.
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| 15. |
- Waldie, Sarah, et al.
(författare)
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Lipoprotein ability to exchange and remove lipids from model membranes as a function of fatty acid saturation and presence of cholesterol.
- 2020
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Ingår i: Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids. - : Elsevier. - 1388-1981 .- 1879-2618. ; 1865:10
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Tidskriftsartikel (refereegranskat)abstract
- Lipoproteins play a central role in the development of atherosclerosis. High and low-density lipoproteins (HDL and LDL), known as 'good' and 'bad' cholesterol, respectively, remove and/or deposit lipids into the artery wall. Hence, insight into lipid exchange processes between lipoproteins and cell membranes is of particular importance in understanding the onset and development of cardiovascular disease. In order to elucidate the impact of phospholipid tail saturation and the presence of cholesterol in cell membranes on these processes, neutron reflection was employed in the present investigation to follow lipid exchange with both HDL and LDL against model membranes. Mirroring clinical risk factors for the development of atherosclerosis, lower exchange was observed in the presence of cholesterol, as well as for an unsaturated phospholipid, compared to faster exchange when using a fully saturated phospholipid. These results highlight the importance of membrane composition on the interaction with lipoproteins, chiefly the saturation level of the lipids and presence of cholesterol, and provide novel insight into factors of importance for build-up and reversibility of atherosclerotic plaque. In addition, the correlation between the results and well-established clinical risk factors suggests that the approach taken can be employed also for understanding a broader set of risk factors including, e.g., effects of triglycerides and oxidative stress, as well as local effects of drugs on atherosclerotic plaque formation.
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| 16. |
- Abletshauser, C, et al.
(författare)
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Biosensing of arteriosclerotic nanoplaque formation and interaction with an HMG-CoA reductase inhibitor
- 2002
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Ingår i: Acta Physiologica Scandinavica. - 0001-6772 .- 1365-201X. ; 176, s. 131-146
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Tidskriftsartikel (refereegranskat)abstract
- Proteoheparan sulphate can be adsorbed to a methylated silica surface in a monomolecular layer via its transmembrane hydrophobic protein core domain. As a result of electrostatic repulsion, its anionic glycosaminoglycan side chains are stretched out into the blood substitute solution, thereby representing one receptor site for specific lipoprotein binding through basic amino acid-rich residues within their apolipoproteins. The binding process was studied by ellipsometric techniques suggesting that high-density lipoprotein (HDL) has a high binding affinity and a protective effect on interfacial heparan sulphate proteoglycan layers with respect to low-density lipoprotein (LDL) and Ca2+ complexation. Low-density lipoprotein was found to deposit strongly at the proteoheparan sulphate-coated surface, particularly in the presence of Ca2+, apparently through complex formation 'proteoglycan-LDL-calcium'. This ternary complex build-up may be interpreted as arteriosclerotic nanoplaque formation on the molecular level responsible for the arteriosclerotic primary lesion. On the other hand, HDL bound to heparan sulphate proteoglycan protected against LDL deposition and completely suppressed calcification of the proteoglycan-lipoprotein complex. In addition, HDL was able to decelerate the ternary complex deposition. Therefore, HDL attached to its proteoglycan receptor sites is thought to raise a multidomain barrier, selection and control motif for transmembrane and paracellular lipoprotein uptake into the arterial wall. Although much remains unclear regarding the mechanism of lipoprotein depositions at proteoglycan-coated surfaces, it seems clear that the use of such systems offers possibilities for investigating lipoprotein deposition at a 'nanoscopic' level under close to physiological conditions. In particular, Ca2+-promoted LDL deposition and the protective effect of HDL even at high Ca2+ and LDL concentrations agree well with previous clinical observations regarding risk and beneficial factors for early stages of atherosclerosis. Considering this, the system was tested on its reliability in a biosensor application in order to unveil possible acute pleiotropic effects of the lipid lowering drug fluvastatin. The very low-density lipoprotein (VLDL)/intermediate-density lipoprotein (IDL)/LDL plasma fraction from a high risk patient with dyslipoproteinaemia and type 2 diabetes mellitus showed beginning arteriosclerotic nanoplaque formation already at a normal blood Ca2+ concentration, with a strong increase at higher Ca2+ concentrations. Fluvastatin, whether applied to the patient (one single 80 mg slow release matrix tablet) or acutely in the experiment (2.2 μmol L-1), markedly slowed down this process of ternary aggregational nanoplaque complexation at all Ca2+ concentrations used. This action resulted without any significant change in lipid concentrations of the patient. Furthermore, after ternary complex build-up, fluvastatin, similar to HDL, was able to reduce nanoplaque adsorption and size. These immediate effects of fluvastatin have to be taken into consideration while interpreting the clinical outcome of long-term studies.
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| 17. |
- Burman, Robert, et al.
(författare)
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Cyclotide-membrane interactions: defining factors of membrane binding, depletion and disruption
- 2011
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Ingår i: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736 .- 1879-2642. ; 1808:11, s. 2665-2673
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Tidskriftsartikel (refereegranskat)abstract
- The cyclotide family of plant-derived peptides is defined by a cyclic backbone and three disulfide bonds locked into a cyclic cystine knot. They display a diverse range of biological activities, many of which have been linked to an ability to target biological membranes. In the current work, we show that membrane binding and disrupting properties of prototypic cyclotides are dependent on lipid composition, using neutral (zwitterionic) membranes with or without cholesterol and/or anionic lipids. Cycloviolacin O2 (cyO2) caused potent membrane disruption, and showed selectivity towards anionic membranes, whereas kalata B1 and kalata B2 cyclotides were significantly less lytic towards all tested model membranes. To investigate the role of the charged amino acids of cyO2 in the membrane selectivity, these were neutralized using chemical modifications. In contrast to previous studies on the cytotoxic and antimicrobial effects of these derivatives, the Glu6 methyl ester of cyO2 was more potent than the native peptide. However, using membranes of Escherichia coil lipids gave the opposite result: the activity of the native peptide increased 50-fold. By using a combination of ellipsometry and LC-MS, we demonstrated that this unusual membrane specificity is due to native cyO2 extracting preferentially phosphatidylethanolamine-lipids from the membrane, i.e., PE-C16:0/cyC17:0 and PE-C16:0/C18:1.
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| 18. |
- Burman, Robert, 1979-, et al.
(författare)
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Membrane integrity as a target for cyclotide cytotoxic activity
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- The cyclotides are a family of plant-derived proteins that occur in plants from the Violaceae (violet) and Rubiaceae (coffee) families and have a diverse range of biological activities, including cytotoxic, hemolytic, antimicrobial, and insecticidal activities; the latter suggests their natural function lies in plant defense. In the current study we have investigated the membrane-disrupting and adsorption ability of prototypic cyclotides and correlated these findings to their cytotoxic properties. We also included modifications of selected charged amino acids in cycloviolacin O2, previously shown to be of importance for its cytotoxic activity. The cyclotides’ cytotoxic activity, ability to adsorb and disrupt model lipid membranes of different charge densities was investigated, e.g. by fluorescence spectroscopy, ellipsometry, and circular dichroism. Cytotoxicity of the native cyclotides was demonstrated to correlate to membrane adsorption and lytic activity. Hence, the activity of native cyclotides is mainly due to interactions between the proteins and the phospholipids in the target membrane. Striking effects of single amino acid variations in cycloviolacin O2 on its membrane interaction were also demonstrated.
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| 19. |
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| 20. |
- Hahn Berg, IC, et al.
(författare)
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Salivary protein adsorption onto hydroxyapatite and SDS-mediated elution studied by in situ ellipsometry
- 2001
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Ingår i: Biofouling (Print). - 0892-7014 .- 1029-2454. ; 17, s. 173-187
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Tidskriftsartikel (refereegranskat)abstract
- Whole unstimulated saliva from two donors was investigated both with respect to adsorption characteristics and SDS-induced elutability. Salivary protein adsorption onto hydroxyapatite (HA) discs was studied by means of in situ ellipsometry in the concentration range 0.1-20% saliva. The adsorbed amounts on HA were found to be similar to those on silica, but the rates of adsorption were lower. Protein adsorption was virtually unaffected by the presence of Na+, whereas Ca2+ induced nucleation of calcium phosphate at the surface, the deposition rate being influenced by the pellicle age but not by the presence of saliva in bulk solution. The SDS elutability of adsorbed pellicles was determined on HA as well as on silica surfaces. Desorption from both surfaces was found to occur in the same SDS concentration range, although a residual layer was observed on HA. The slight net positive charge and lower charge density of HA as compared to the strongly negatively charged silica, may, at least partly, account for this observation by causing a reduction in the repulsive force between protein-surfactant complexes and the surface. Interindividual differences, observed in the adsorption as well as elution experiments, are thought to relate to the compositional differences observed by SDS-PAGE
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| 21. |
- Holdbrook, Daniel A., et al.
(författare)
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Influence of pH on the activity of thrombin-derived antimicrobial peptides
- 2018
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Ingår i: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736 .- 1879-2642. ; 1860:11, s. 2374-2384
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Tidskriftsartikel (refereegranskat)abstract
- The wound environment is characterized by physiological pH changes. Proteolysis of thrombin by wound-derived proteases, such as neutrophil elastase, generates antimicrobial thrombin-derived C-terminal peptides (TCPs), such as HVF18 (HVFRLKKWIQKVIDQFGE). Presence of such TCPs in human wound fluids in vivo, as well as the occurrence of an evolutionarily conserved His residue in the primary amino acid sequence of TCPs, prompted us to investigate the pH-dependent antibacterial action of HVF18, as well as of the prototypic GKY25 (GKYGFYTHVFRLKKWIQKVIDQFGE). We show that protonation of this His residue at pH 5.5 increases the antibacterial activity of both TCPs against Gram-negative Escherichia coli by membrane disruption. Physiological salt level (150 mM NaCl) augments antibacterial activity of GKY25 but diminishes for the shorter HVF18. Replacing His with Leu or Ser in GKY25 abolishes the His protonation-dependent increase in antibacterial activity at pH 5.5, whereas substitution with Lys maintains activity at neutral (pH 7.4) and acidic pH. Interestingly, both TCPs display decreased binding affinities to human CD14 with decreasing pH, suggesting a likely switch in mode-of-action, from anti-inflammatory at neutral pH to antibacterial at acidic pH. Together, the results demonstrate that apart from structural prerequisites such as peptide length, charge, and hydrophobicity, the evolutionarily conserved His residue of TCPs influences their antibacterial effects and reveals a previously unknown aspect of TCPs biological action.
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| 22. |
- Häffner, Sara Malekkhaiat, et al.
(författare)
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Membrane Interactions of Virus-like Mesoporous Silica Nanoparticles
- 2021
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Ingår i: ACS Nano. - : American Chemical Society (ACS). - 1936-0851 .- 1936-086X. ; 15:4, s. 6787-6800
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, we investigated lipid membrane interactions of silica nanoparticles as carriers for the antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES). In doing so, smooth mesoporous nanoparticles were compared to virus-like mesoporous nanoparticles, characterized by a "spiky"external surface, as well as to nonporous silica nanoparticles. For this, we employed a combination of neutron reflectometry, ellipsometry, dynamic light scattering, and ζ-potential measurements for studies of bacteria-mimicking bilayers formed by palmitoyloleoylphosphatidylcholine/palmitoyloleoylphosphatidylglycerol. The results show that nanoparticle topography strongly influences membrane binding and destabilization. We found that virus-like particles are able to destabilize such lipid membranes, whereas the corresponding smooth silica nanoparticles are not. This effect of particle spikes becomes further accentuated after loading of such particles with LL-37. Thus, peptide-loaded virus-like nanoparticles displayed more pronounced membrane disruption than either peptide-loaded smooth nanoparticles or free LL-37. The structural basis of this was clarified by neutron reflectometry, demonstrating that the virus-like nanoparticles induce trans-membrane defects and promote incorporation of LL-37 throughout both bilayer leaflets. The relevance of such effects of particle spikes for bacterial membrane rupture was further demonstrated by confocal microscopy and live/dead assays on Escherichia coli bacteria. Taken together, these findings demonstrate that topography influences the interaction of nanoparticles with bacteria-mimicking lipid bilayers, both in the absence and presence of antimicrobial peptides, as well as with bacteria. The results also identify virus-like mesoporous nanoparticles as being of interest in the design of nanoparticles as delivery systems for antimicrobial peptides.
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| 23. |
- Ilyas, Humaira, et al.
(författare)
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Effect of PEGylation on Host Defense Peptide Complexation with Bacterial Lipopolysaccharide
- 2021
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Ingår i: Bioconjugate Chemistry. - : American Chemical Society (ACS). - 1043-1802 .- 1520-4812. ; 32:8, s. 1729-1741
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Tidskriftsartikel (refereegranskat)abstract
- Conjugation with poly(ethylene glycol) ("PEGylation") is a widely used approach for improving the therapeutic propensities of peptide and protein drugs through prolonging bloodstream circulation, reducing toxicity and immunogenicity, and improving proteolytic stability. In the present study, we investigate how PEGylation affects the interaction of host defense peptides (HDPs) with bacterial lipopolysaccharide (LPS) as well as HDP suppression of LPS-induced cell activation. In particular, we investigate the effects of PEGylation site for KYE28 (KYEITTIHNLFRKLTHRLFRRNFGYTLR), a peptide displaying potent anti-inflammatory effects, primarily provided by its N-terminal part. PEGylation was performed either in the N-terminus, the C-terminus, or in both termini, keeping the total number of ethylene groups (n = 48) constant. Ellipsometry showed KYE28 to exhibit pronounced affinity to both LPS and its hydrophobic lipid A moiety. The PEGylated peptide variants displayed lower, but comparable, affinity for both LPS and lipid A, irrespective of the PEGylation site. Furthermore, both KYE28 and its PEGylated variants triggered LPS aggregate disruption. To investigate the peptide structure in such LPS complexes, a battery of nuclear magnetic resonance (NMR) methods was employed. From this, it was found that KYE28 formed a well-folded structure after LPS binding, stabilized by hydrophobic domains involving aromatic amino acids as well as by electrostatic interactions. In contrast, the PEGylated peptide variants displayed a less well-defined secondary structure, suggesting weaker LPS interactions in line with the ellipsometry findings. Nevertheless, the N-terminal part of KYE28 retained helix formation after PEGylation, irrespective of the conjugation site. For THP1-Xblue-CD14 reporter cells, KYE28 displayed potent suppression of LPS activation at simultaneously low cell toxicity. Interestingly, the PEGylated KYE28 variants displayed similar or improved suppression of LPS-induced cell activation, implying the underlying key role of the largely retained helical structure close to the N-terminus, irrespective of PEGylation site. Taken together, the results show that PEGylation of HDPs can be done insensitively to the conjugation site without losing anti-inflammatory effects, even for peptides inducing such effects through one of its termini.
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| 24. |
- Joiner, A, et al.
(författare)
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Adsorption from black tea and red wine onto in vitro salivary pellicles studied by ellipsometry
- 2003
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Ingår i: European Journal of Oral Sciences. - : Blackwell Munksgaard. - 0909-8836 .- 1600-0722. ; 111, s. 417-422
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Tidskriftsartikel (refereegranskat)abstract
- The adsorption of black tea and red wine components onto a pellicle-like protein layer formed in vitro by adsorption from whole unstimulated saliva on hydroxyapatite discs were studied by in situ ellipsometry. It was found that components from black tea readily adsorbed to the pellicle. Subsequent exposure to saliva led to further adsorption of salivary components to give an overall increase in the amounts adsorbed. The amounts adsorbed increased still further following a third tea and saliva exposure. Components of red wine gave significantly greater amounts of adsorption to the pellicle than black tea. The adsorption of components of black tea gave a concomitant increase in colour or stain as measured by a reflectance chromameter. In all cases, the black tea- and red wine-modified pellicles were not eluted by either phosphate buffer or sodium dodecyl sulphate (SDS) rinses. Thus, black tea and red wine components have been shown to have a profound effect on in vitro pellicle maturation, causing thickened layers of stained material to build up, which are not readily removed.
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| 25. |
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| 26. |
- Kapilashrami, A, et al.
(författare)
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Drying of oil-in-water emulsions on hydrophobic and hydrophilic substrates
- 2004
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Ingår i: Colloids and Surfaces A. - 0927-7757 .- 1873-4359. ; 233, s. 155-161
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Tidskriftsartikel (refereegranskat)abstract
- We have investigated the effect of the hydrophobic/hydrophilic character of the substrates on the drying behaviour of dilute silicone oil-in-water (o/w) emulsions by light microscopy and ellipsometry. The poly(dimethylsiloxane) (PDMS) emulsion droplets, which are stabilised by a triblock PEO/PPO/PEO copolymer, form a close-packed structure containing domains of hexagonally packed droplets on the hydrophilic substrate. We find that the hydrophilic substrate does not destabilise the emulsion droplets; the close-packed structures are very stable and coalesce very slowly only when most of the water has evaporated. This is supported by ellipsometry measurements, which showed that the emulsion droplets do not adsorb to the hydrophilic substrate. The hydrophobic substrate, on the other hand, destabilises the emulsion and we observed a significant increase in the coalescence rate. Ellipsometry measurements suggest that destabilisation is promoted by the strong interaction between the emulsion droplets and the hydrophobic substrate. We also find that the emulsion undergoes dewetting followed by a release of oil and rewetting of the substrate when the o/w emulsion film reaches a critical thickness on the hydrophobic substrate
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| 27. |
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| 28. |
- Kapilashrami, A, et al.
(författare)
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Ellipsometric studies of nonionic block copolymers adsorbed at the solid/water and oil/water interfaces
- 2003
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Ingår i: Colloids ad Surfaces a-Physicochemical and Engineering Aspects. ; 225:1-3, s. 181-192
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Tidskriftsartikel (refereegranskat)abstract
- We report on the interfacial behaviour of a series of nonionic diblock copolymers at solid hydrophobic and hydrophilic surfaces/water and silicone oil/water interfaces, studied by ellipsometry. The polymers consist of a hydrophobic C-18 chain linked to a hydrophilic poly(ethylene oxide) (PEO), block varying from 50 to 250 U. The adsorption of these copolymers at low bulk concentrations was found to be dominated by the PEO block at all interfaces. At higher concentration the copolymer forms surface aggregates at the silica surface whereas we observe a gradual increase in the adsorbed layer thickness with increased surface excess at the solid hydrophobic surface, indicating a transition from a flat conformation to brush-like layer structure. The results indicate a similar evolution in adsorbed amount with concentration at the silicone oil/water interface as at the hydrophobic silica surface. The influence of the rheological proper ies of the interface on the adsorption of the diblock copolymer was investigated by comparing results from two silicon oils with different viscosities. The copolymers were found to have stronger affinity to a low viscosity (990 mPa s) silicone oil than to a higher viscosity (12 800 mPa s) silicone oil and the hydrophobised silica surface. At the silicone oil/water interface the adsorption of a commercial nonionic triblock copolymer was furthermore investigated and compared with the diblock copolymers. (C) 2003 Elsevier B.V. All rights reserved.
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| 29. |
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| 30. |
- Kapilashrami, A., et al.
(författare)
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Ellipsometric studies of nonionic copolymers adsorbed at the solid/water and oil/water interfaces
- 2003
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Ingår i: Colloids and Surfaces A. - 0927-7757 .- 1873-4359. ; 225, s. 181-192
-
Tidskriftsartikel (refereegranskat)abstract
- We report on the interfacial behaviour of a series of nonionic diblock copolymers at solid hydrophobic and hydrophilic surfaces/water and silicone oil/water interfaces, studied by ellipsometry. The polymers consist of a hydrophobic C18 chain linked to a hydrophilic poly(ethylene oxide) (PEO), block varying from 50 to 250 U. The adsorption of these copolymers at low bulk concentrations was found to be dominated by the PEO block at all interfaces. At higher concentration the copolymer forms surface aggregates at the silica surface whereas we observe a gradual increase in the adsorbed layer thickness with increased surface excess at the solid hydrophobic surface, indicating a transition from a flat conformation to brush-like layer structure. The results indicate a similar evolution in adsorbed amount with concentration at the silicone oil/water interface as at the hydrophobic silica surface. The influence of the rheological properties of the interface on the adsorption of the diblock copolymer was investigated by comparing results from two silicon oils with different viscosities. The copolymers were found to have stronger affinity to a low viscosity (990 mPa s) silicone oil than to a higher viscosity (12800 mPa s) silicone oil and the hydrophobised silica surface. At the silicone oil/water interface the adsorption of a commercial nonionic triblock copolymer was furthermore investigated and compared with the diblock copolymers
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| 31. |
- Lin, Jay, et al.
(författare)
-
High prevalence of hepatitis E virus in Swedish moose : A phylogenetic characterization and comparison of the virus from different regions
- 2015
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 10:4
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Tidskriftsartikel (refereegranskat)abstract
- Background: Hepatitis E virus (HEV) infects a range of species, including humans, pigs, wild boars and deer. Zoonotic transmission may contribute to the high HEV seroprevalence in the human population of many countries. A novel divergent HEV from moose (Alces alces) in Sweden was recently identified by partial genome sequencing. Since only one strain was found, its classification within the HEV family, prevalence in moose and zoonotic potential was unclear. We therefore investigated samples from 231 moose in seven Swedish counties for HEV, and sequenced a near complete moose HEV genome. Phylogenetic analysis to classify this virus within the family Hepeviridae and to explore potential host specific determinants was performed. Methods and Findings: The HEV prevalence of moose was determined by PCR (marker for active infection) and serological assays (marker of past infection) of sera and 51 fecal samples from 231 Swedish moose. Markers of active and past infection were found in 67 (29%) animals, while 34 (15%) were positive for HEV RNA, 43 (19%) were seropositive for anti-HEV antibodies, and 10 (4%) had both markers. The number of young individuals positive for HEV RNA was larger than for older individuals, and the number of anti-HEV antibody positive individuals increased with age. The high throughput sequenced moose HEV genome was 35-60% identical to existing HEVs. Partial ORF1 sequences from 13 moose strains showed high similarity among them, forming a distinct monophyletic clade with a common ancestor to HEV genotype 1-6 group, which includes members known for zoonotic transmission. Conclusions: This study demonstrates a high frequency of HEV in moose in Sweden, with markers of current and past infection demonstrated in 30% of the animals. Moose is thus an important animal reservoir of HEV. The phylogenetic relationship demonstrated that the moose HEV belonged to the genotype 1-6 group, which includes strains that also infect humans, and therefore may signify a potential for zoonotic transmission of this HEV. © 2015 Lin et al.
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| 32. |
- Lindström, A, et al.
(författare)
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Interactions in phospholipid stabilized emulsion system
- 1999
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Ingår i: Journal of Dispersion Science and Technology. - 0193-2691 .- 1532-2351. ; 20, s. 247-256
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Tidskriftsartikel (refereegranskat)abstract
- The interaction between a phospholipid stabilized triglyceride emulsion and a hydrophilic silica surface has been studied at varying pH and electrolyte content using ellipsometry. The adsorbed amount decreases with pH and increases with increasing electrolyte content in the emulsion, and this can be rationalized on the basis of the electrostatic interaction between the emulsion droplet and the surface. The layer thickness, however, is essentially independent of these parameters. In addition, the emulsion has been studied during turbulent shear conditions (applied mechanical stress), with the same variation of pH and electrolyte as in the adsorption experiments. A decrease in pH and an increase in electrolyte content, decreasing the repulsive interaction between the droplets, leads to a deterioration in emulsion stability with time.
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| 33. |
- Malekkhaiat Häffner, S., et al.
(författare)
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Composition effects on photooxidative membrane destabilization by TiO2 nanoparticles
- 2021
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797. ; 584, s. 19-33
-
Tidskriftsartikel (refereegranskat)abstract
- Membrane interactions and photooxidative membrane destabilization of titanium dioxide (TiO2) nanoparticles were investigated, focusing on the effects of membrane composition, notably phospholipid headgroup charge and presence of cholesterol. For this, we employed a battery of state-of-the-art methods for studies of bilayers formed by zwitterionic palmitoyloleoylphosphatidylcholine (POPC) containing also polyunsaturated palmitoylarachidonoylphosphocholine (PAPC), as well as its mixtures with anionic palmitoyloleoylphosphatidylglycerol (POPG) and cholesterol. It was found that the TiO2 nanoparticles display close to zero charge at pH 7.4, resulting in aggregation. At pH 3.4, in contrast, the 6 nm TiO2 nanoparticles are well dispersed due to a strongly positive ζ-potential. Mirroring this pH dependence, TiO2 nanoparticles were observed to bind to negatively charged lipid bilayers at pH 3.4, but much less so at pH 7.4. While nanoparticle binding has some destabilizing effect alone, illumination with ultraviolet (UV) light accentuates membrane destabilization, a result of oxidative stress caused by generated reactive oxygen species (ROS). Neutron reflectivity (NR), quartz crystal microbalance (QCM), and small-angle X-ray scattering (SAXS) results all demonstrate that membrane composition strongly influences membrane interactions and photooxidative destabilization of lipid bilayers. In particular, the presence of anionic POPG makes the bilayers more sensitive to oxidative destabilization, whereas a stabilizing effect was observed in the presence of cholesterol. Also, structural aspects of peroxidation were found to depend strongly on membrane composition, notably the presence of anionic phospholipids. The results show that membrane interactions and UV-induced ROS generation act in concert and need to be considered together to understand effects of lipid membrane composition on UV-triggered oxidative destabilization by TiO2 nanoparticles, e.g., in the context of oxidative damage of bacteria and cells.
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| 34. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Membrane interactions and antimicrobial effects of layered double hydroxide nanoparticles
- 2017
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Ingår i: Physical Chemistry, Chemical Physics - PCCP. - : ROYAL SOC CHEMISTRY. - 1463-9076 .- 1463-9084. ; 19:35, s. 23832-23842
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Tidskriftsartikel (refereegranskat)abstract
- Membrane interactions are critical for the successful use of inorganic nanoparticles as antimicrobial agents and as carriers of, or co-actives with, antimicrobial peptides (AMPs). In order to contribute to an increased understanding of these, we here investigate effects of particle size (42-208 nm) on layered double hydroxide (LDH) interactions with both bacteria-mimicking and mammalian-mimicking lipid membranes. LDH binding to bacteria-mimicking membranes, extraction of anionic lipids, as well as resulting membrane destabilization, was found to increase with decreasing particle size, also translating into size-dependent synergistic effects with the antimicrobial peptide LL-37. Due to strong interactions with anionic lipopolysaccharide and peptidoglycan layers, direct membrane disruption of both Gram-negative and Gram-positive bacteria is suppressed. However, LDH nanoparticles cause size-dependent charge reversal and resulting flocculation of both liposomes and bacteria, which may provide a mechanism for bacterial confinement or clearance. Taken together, these findings demonstrate a set of previously unknown behaviors, including synergistic membrane destabilization and dual confinement/killing of bacteria through combined LDH/AMP exposure, of potential therapeutic interest.
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| 35. |
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| 36. |
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| 37. |
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| 38. |
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| 39. |
- Malmsten, A, et al.
(författare)
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Temperature-dependent adsorption and surface forces in aqueous ethyl(hydroxyethyl)cellulose solutions
- 1991
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Ingår i: Langmuir. - 0743-7463 .- 1520-5827. ; 7, s. 988-994
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Tidskriftsartikel (refereegranskat)abstract
- Forces between hydrophobized mica surfaces across an aqueous solution of ethyl(hydroxyethyl)cellulose (EHEC) have been studied, inter alia, as a function of surface separation and temperature. EHEC adsorbs strongly to hydrophobic surfaces in a rather flat conformation. The force is reversible on approach and separation up to a temperature of about 40°C. As the temperature was increased from 20°C to 37°C, which is slightly above the cloud point (CP=35°C), the adsorbed amount increased by a factor of three. The force remained monotonically repulsive, despite the fact that the temperature was higher than CP. On increasing the temperature further, the adsorbed amount, as well as the thickness of the adsorbed layer (at a high compressional force), remains essentially unchanged. However, due to the decreased solvency, there is a contraction of the outer part of the adsorbed layer. The force was essentially repulsive at 41°C, with only a very weak attraction observed on separating the surfaces. At 52°C, a weak attraction was observed on both approach and separation, as would be expected in a significantly worse than q-solvent. As the temperature was decreased again, the adsorbed amount as well as the compressed adsorbed layer thickness decreased. The adsorbed amount was the same (within experimental uncertainty) after heating as before. However, the force curve was shifted outwards after heating as compared to the situation before heating. Hence, the temperature dependence of the adsorption, in the absence of constraints on the adsorbed amount, is not completely reversible, at least over the incubation time used (12 h).
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| 40. |
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| 41. |
- Malmsten, M, et al.
(författare)
-
A model substrate for ellipsometry studies of lipoprotein deposition at the endothelium
- 2001
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 240, s. 372-374
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Tidskriftsartikel (refereegranskat)abstract
- Endothelial cell layers from umbilical cords were grown on methylated silica surfaces. Fluorescence microscopy showed this layer to be confluent and to consist of living cells. Initial ellipsometry measurements were performed to illustrate both the stability of the model surface in ellipsometry measurements and the Ca2+-dependent binding of lipoproteins at the cell-based substrate. The present substrate holds promise as a model substrate for in vitro investigations of lipoprotein deposition at the endothelium surface under close to in vivo conditions.
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| 42. |
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| 43. |
- Malmsten, M, et al.
(författare)
-
Effects of amino acid composition on protein adsorption
- 1996
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 178, s. 160-167
-
Tidskriftsartikel (refereegranskat)abstract
- The adsorption at silica and methylated silica of genetically engineered derivatives of Z and its dimer ZZ, where Z is a hydrophilic synthetic IgG binding domain derived from staphylococcal protein A, was studied with in situ ellipsometry. The protein modifications consisted of introducing short peptide stretches near the C-terminus of the protein. Using this approach, oligopeptide stretches containing hydrophobic tryptophan (Trp) or isoleucine (Ile) [(AlaTrpTrpPro)n or (AlaIleIlePro)n (0≤n≤2), denoted Tn and In, respectively] were introduced in the protein. For comparison, the adsorption of the inserted peptide stretches (Tn and In), as well as of Trp and Ile oligomers, was investigated as well. Increasing the number of Trp residues resulted in an increased adsorption for both ZZTn, ZTn, Tn, and Trpn. At a given number of Trp residues, the adsorbed amount of the ZZ derivatives is larger than that of the peptides, but about the same as that of the Z proteins. Analogous although somewhat smaller effects were obtained for the Ile-derivatized proteins. These results are discussed in terms of the "block copolymer" nature of the proteins. Theoretical calculations using a mean-field lattice model for block copolymer adsorption gave a qualitative agreement with the experimentally obtained results.
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| 44. |
- Malmsten, M, et al.
(författare)
-
Electrostatic and hydrophobic effects of oligopeptide insertions on protein adsorption
- 1998
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 204, s. 104-111
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Tidskriftsartikel (refereegranskat)abstract
- The effects of oligopeptide insertions on the adsorption of the protein ZZ, where Z is the IgG binding domain of staphylococcal Protein A, was investigated by in situ ellipsometry. In particular, the interplay between hydrophobic and electrostatic interactions as driving force for adsorption was investigated by studying the effects of oligopeptide insertions of the type Tn(AlaTrpTrpPro), Na ((AlaTrpTrpLysPro)n), and Pn((AlaTrpTrpLysPro)n) on the adsorption at silica, methylated silica , and diaminocyclohexane (DACH) plasma polymer surfaces. For comparison, the adsorption of the inserted peptide stretches was also investigated. It was found that the adsorption of all the peptides increases with the molecular weight at methylated silica. At silica, only the Pn peptides were found to adsorb. The net negatively charged proteins modified through peptide insertions did not adsorb at the hydrophilic and negatively charged silica, irrespective of the peptide insertion, whereas an extensive adsorption was found for the posetively charged DACH surface for all the proteins investigated. For hydrophobic and negatively charged methylated silica, on the other hand, the peptide insertions were found to have a major influence on the protein interfacial behavior, and the adsorption followed the peptide stretch charge, thus increasing in the order ZZNn< ZZTn < ZZPn. These effects are discussed in terms of the relative importance of hydrophobic and electrostatic interactions as driving force for the adsorption.
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| 45. |
- Malmsten, M, et al.
(författare)
-
Electrostatic effects on interfacial film formation in emulsion systems
- 1996
-
Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 179, s. 537-543
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Tidskriftsartikel (refereegranskat)abstract
- The adsorption at silica from dilute emulsion systems was studied with in situ ellipsometry. In particular, the effects of electrostatic interactions on the adsorption rate and the adsorbed layer structure and formation was investigated by varying the emulsion droplet and surface charge, as well as the electrostatic screening, accomplished by varying pH and the excess electrolyte concentration. Electrostatic interactions were found to markedly affect the adsorption rate, but not the adsorbed layer structure or the mechanism for the adsorbed layer formation. For all cases investigated, the adsorbed layer thickness corresponds to emulsion droplets or multilamellar liposomes, and the adsorbed layer formation proceeds through attachment of emulsion droplets and/or multilamellar liposomes at the surface without extensive droplet spreading or liposome collapse. When the droplets and the surface are similarly charged, the adsorption is facilitated by increasing the electrostatic screening or by decreasing the emulsion droplet and surface charge, accomplished by increasing the excess electrolyte concentration and decreasing pH, respectively. When the droplets and the surface are oppositely charged, the adsorption rate is much higher than that observed when the droplets and the surface are similarly charged, although the adsorbed layer structure and the mechanism for the adsorbed layer formation are similar. Qualitatively, these effects may be understood by considering only electrostatic and van der Waals interactions.
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| 46. |
- Malmsten, M, et al.
(författare)
-
Ellipsometry studies of interfacial film formation in emulsion systems
- 1995
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 173, s. 297-303
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Tidskriftsartikel (refereegranskat)abstract
- The possibility of using in situ ellipsometry for studies of the formation of interfacial films in emulsion systems was investigated. It was found that ellipsometry studies on dilute emulsion can provide information on both the adsorption kinetics, the adsorbed layer structure and the mechanisms for the adsorbed layer formation. At high electrolyte concentrations, the adsorption from a model o/w emulsion stabilized by egg lecithin was found to be more pronounced at hydrophilic and negatively charged silica than at hydrophobic methylated silica. Furthermore, the adsorbed layer thickness at both surfaces is smaller than the average droplet size, corresponding to either small or ”flattened” droplets or liposomes. The adsorbed layer thickness is smaller at methylated silica than at silica, presumably due to a larger degree of emulsion droplet or liposome spreading at this surface. In both cases, the adsorbed layer thickness is independent of the adsorbed amount, i. e. the adsorbed layer formation proceeds by attachment of droplets, which pack more densly with an increasing adsorbed amount.
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| 47. |
- Malmsten, M, et al.
(författare)
-
Immobilization of trypsin on porous glycidyl methacrylate beads; effects of polymer hydrophilization
- 2000
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Ingår i: Colloids and Surfaces B. - 0927-7765 .- 1873-4367. ; 18, s. 277-284
-
Tidskriftsartikel (refereegranskat)abstract
- The immobilization of trypsin at porous glycidyl methacrylate (GMA-GDMA) beads was investigated. In particular, the effects of surface modification of the beads through hydrophilic polymers on the amount protein immobilized and on the extent of retained activity after immobilization were adressed. Furthermore, immobilization at unmodified and hydrophilized beads from aqueous solution was compared to that from a water-in-oil microemulsion. It was found that the amount trypsin immobilized at the unmodified GMA-GDMA beads was significantly higher than that at hydrophilized GMA-GDMA beads. However, also the extent of specific activity loss after immobilization was larger for the unmodified than for the hydrophilized beads. Despite the latter, however, the total activity displayed by the hydrophilized beads was comparable to the unmodified beads at best. On the other hand, by peforming the immobilization from the microemulsion a high immobilization yield can be reached even for the hydrophilized beads, which also results in a higher degree of retained activity in the latter case than obtained for immobilization at the unmodified beads. Using this approach therefore resulted in the highest total activity of the trypsin-activated GMA-GDMA beads.
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| 48. |
- Martin, Alma, et al.
(författare)
-
In Situ Transformation of Electrospun Nanofibers into Nanofiber-Reinforced Hydrogels
- 2022
-
Ingår i: Nanomaterials. - : MDPI AG. - 2079-4991. ; 12:14
-
Tidskriftsartikel (refereegranskat)abstract
- Nanofiber-reinforced hydrogels have recently gained attention in biomedical engineering. Such three-dimensional scaffolds show the mechanical strength and toughness of fibers while benefiting from the cooling and absorbing properties of hydrogels as well as a large pore size, potentially aiding cell migration. While many of such systems are prepared by complicated processes where fibers are produced separately to later be embedded in a hydrogel, we here provide proof of concept for a one-step solution. In more detail, we produced core-shell nanofibers from the natural proteins zein and gelatin by coaxial electrospinning. Upon hydration, the nanofibers were capable of directly transforming into a nanofiber-reinforced hydrogel, where the nanofibrous structure was retained by the zein core, while the gelatin-based shell turned into a hydrogel matrix. Our nanofiber-hydrogel composite showed swelling to ~800% of its original volume and water uptake of up to ~2500% in weight. The physical integrity of the nanofiber-reinforced hydrogel was found to be significantly improved in comparison to a hydrogel system without nanofibers. Additionally, tetracycline hydrochloride was incorporated into the fibers as an antimicrobial agent, and antimicrobial activity against Staphylococcus aureus and Escherichia coli was confirmed.
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| 49. |
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| 50. |
- Nordström, Randi, 1986-, et al.
(författare)
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Degradable dendritic nanogels as carriers for antimicrobial peptides
- 2019
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 554, s. 592-602
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, we investigate degradable anionic dendritic nanogels (DNG) as carriers for antimicrobial peptides (AMPs). In such systems, the dendritic part contains carboxylic acid-based anionic binding sites for cationic AMPs, whereas linear poly(ethylene glycol) (PEG) chains form a shell for promotion of biological stealth. In order to clarify factors influencing membrane interactions of such systems, we here address effects of nanogel charge, cross-linking, and degradation on peptide loading/release, as well as consequences of these factors for lipid membrane interactions and antimicrobial effects. The DNGs were found to bind the AMPs LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) and DPK-060 (GKHKNKGKKNGKHNGWKWWW). For the smaller DPK-060 peptide, loading was found to increase with increasing nanogel charge density. For the larger LL-37, on the other hand, peptide loading was largely insensitive to nanogel charge density. In line with this, results on the secondary structure, as well as on the absence of stabilization from proteolytic degradation by the nanogels, show that the larger LL-37 is unable to enter into the interior of the nanogels. While 40–60% nanogel degradation occurred over 10 days, promoted at high ionic strength and lower cross-linking density/higher anionic charge content, peptide release at physiological ionic strength was substantially faster, and membrane destabilization not relying on nanogel degradation. Ellipsometry and liposome leakage experiments showed both free peptide and peptide/DNG complexes to cause membrane destabilization, indicated also by antimicrobial activities being comparable for nanogel-bound and free peptide. Finally, the DNGs were demonstrated to display low toxicity towards erythrocytes even at peptide concentrations of 100 µM.
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