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1.
Lipid layers on polyelectrolyte multilayer supports
Martin Fischlechner, Markus Zaulig, Stefan Meyer, Irina Estrela-Lopis, Luis Cuéllar, Joseba Irigoyen, Paula Pescador, Milan Brumen, Paul Messner, Sergio Moya, Edwin Donath, 2008, original scientific article

Abstract: The mechanism of formation of supported lipid layers from phosphatidylcholine and phosphatidylserine vesicles in solution on polyelectrolyte multilayers was studied by a variety of experimental techniques. The interaction of zwitterionic and acidic lipid vesicles, as well as their mixtures, with polyelectrolyte supports was followed in real time by micro-gravimetry. The fabricated lipid-polyelectrolyte composite structures on top of multilayer coated colloidal particles were characterized by flow cytometry and imaging techniques. Lipid diffusion over the macroscopic scale was quantified by fluorescence recovery after photobleaching, and the diffusion was related to layer connectivity. The phospholipid-polyelectrolyte binding mechanism was investigated by infrared spectroscopy. A strong interaction of polyelectrolyte primary amino groups with phosphate and carboxyl groups of the phospholipids, leading to dehydration, was observed. Long-range electrostatic attraction was proven to be essential for vesicle spreading and rupture. Fusion of lipid patches into a homogeneous bilayer required lateral mobility of the lipids on the polyelectrolyte support. The binding of amino groups to the phosphate group of the zwitterionic lipids was too weak to induce vesicle spreading, but sufficient for strong adsorption. Only the mixture of phosphatidylcholine and phosphatidylserine resulted in the spontaneous formation of bilayers on polyelectrolyte multilayers. The adsorption of phospholipids onto multilayers displaying quarternary ammonium polymers produced a novel 3D lipid polyelectrolyte structure on colloidal particles.
Keywords: biophysics, lipids, lipid layers, electrolytes, multilayer supports
Published: 07.06.2012; Views: 790; Downloads: 45
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2.
Flow cytometry of HEK 293T cells interacting with polyelectrolyte multilayer capsules containing fluorescein-labeled poly(acrylic acid) as a pH sensor
Uta Reibetanz, David Haložan, Milan Brumen, Edwin Donath, 2007, original scientific article

Abstract: Polyelectrolyte multilayer sensor capsules, 5 m in diameter, which contained fluorescein-labeled poly(acrylic acid) (PAAAF) as pH-sensitive reporter molecules, were fabricated and employed to explore their endocytotic uptake into HEK 293T cells by flow cytometry. The percentage of capsules residing in the endolysosomal compartment was estimated from the fluorescence intensity decrease caused by acidification. Capsules attached to the extracellular surface of the plasma membrane were identified by trypan blue quenching. The number of capsules in the cytoplasm was rather small, being below the detection limit of the method. The advantages of polyelectrolyte multilayer capsules are that the fluorophore is protected from interaction with cellular compartments and that the multilayer can be equipped with additional functions.
Keywords: biophysics, polyelectrolytes, multilayer capsules, polyelectrolyte capsules
Published: 07.06.2012; Views: 858; Downloads: 81
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3.
Donnan equilibrium and osmotic pressure in hollow polyelectrolyte microcapsules
David Haložan, Gleb B. Sukhorukov, Milan Brumen, Edwin Donath, Helmuth Möhwald, 2007, original scientific article

Abstract: A simple theoretical model of the Donnan equilibrium is applied to studies of distributions of ions between the external and internal volumes of hollow polyelectrolyte capsules as well as of concomitant osmotic pressure load on a capsule wall resulting from these distributions and the presence of polyanion. The model system consist of dispersed polyelectrolyte capsules in an electrolyte solution whereby two cases are considered with respect to the presence of the polyanion, either in the inner or external solution. It is assumed that the capsule wall is impermeable to polyanion, but water and all ions can freely penetrate. The model predictions are summarized by presenting the difference between the external pH, the pH of the inner solution and the osmotic pressure difference across the capsule wall, both in the dependence of sodium chloride concentration.
Keywords: biophysics, polyelectrolytes, polyelectrolyte capsules, Donnan equilibrium, osmotic pressure
Published: 07.06.2012; Views: 1380; Downloads: 73
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4.
Selective regulation of protein activity by complex Ca[sup]2+ oscillations : a theoretical study
Beate Knoke, Marko Marhl, Stefan Schuster, 2007, independent scientific component part or a chapter in a monograph

Abstract: Calcium oscillations play an important role in intracellular signal transduction. As a second messenger, ▫$Ca^{2+}$▫ represents a link between several input signals and several target processes in the cell. Whereas the frequency of simple ▫$Ca^{2+}$▫ oscillations enables a selective activation of a specific protein and herewith a particular process, the question arises of how at the same time two or more classes of proteins can be specifically regulated. The question is general and concerns the problem of how one second messenger can transmit more than one signal simultaneously (bow-tie structure of signalling). To investigate whether a complex ▫$Ca^{2+}$▫ signal like bursting, a succession of low-peak and high-peak oscillatory phases, could selectively activate different proteins, several bursting patterns with simplified square pulses were applied in a theoretical model. The results indicate that bursting ▫$Ca^{2+}$▫ oscillations allow a differential regulation of two different calcium-binding proteins, and hence, perform the desired function.
Keywords: biophysics, calcium oscillations, cellular dynamics, mathematical models, signalling, bow-tie structures, bursting, decoding
Published: 07.06.2012; Views: 1105; Downloads: 12
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5.
Theoretical and experimental investigation of calcium-contraction coupling in airway smooth muscle
Prisca Mbikou, Aleš Fajmut, Milan Brumen, Etienne Roux, 2006, original scientific article

Abstract: We investigated theoretically and experimentally the ▫$Ca^{2+}$▫-contraction couplingin rat tracheal smooth muscle. ▫$[Ca^{2+}]_i$▫, isometric contraction and myosin light chain (MLC) phosphorylation were measured in response to 1 mM carbachol. Theoretical modeling consisted in coupling a model of ▫$Ca^{2+}-dependent$▫ MLC kinase (MLCK) activation with a four-state model of smooth muscle contractile apparatus. Stimulation resulted in a short-time contraction obtained within 1 min, followed by a long-time contraction up to the maximal force obtained in 30 min. ML-7 and Wortmannin (MLCK inhibitors) abolished the contraction. Chelerythrine (PKC inhibitor) did not change the short-time, but reduced the long-time contraction. ▫$[Ca^{2+}]_i$▫ responses of isolated myocytes recorded during the first 90 s consisted in a fast peak, followed by a plateau phase and, in 28 % of the cells, superimposed ▫$Ca^{2+}$▫ oscillations. MLC phosphorylation was maximal at 5 s and then decreased, whereas isometric contraction followed a Hill-shaped curve. The model properlypredicts the time course of MLC phosphorylation and force of the short-time response. With oscillating ▫$Ca^{2+}$▫ signal, the predicted force does not oscillate. According to the model, the amplitude of the plateau and the frequency of oscillations encode for the amplitude of force, whereas the peak encodes for force velocity. The long-time phase of the contraction, associated with a second increase in MLC phosphorylation, may be explained, at least partially, by MLC phosphatase (MLCP) inhibition, possibly via PKC inhibition.
Keywords: biophysics, mathematical modelling, modelling, calcium oscillations, contractions, force development, muscle cells, smooth muscles, myosin kinase
Published: 07.06.2012; Views: 1069; Downloads: 62
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6.
Modeling of molecular and cellular mechanisms involved in [Ca sup 2+] signal encoding in airway myocytes
Marko Marhl, Denis Noble, Etienne Roux, 2006, review article

Abstract: In airway myocytes signal transduction via cytosolic calcium plays an important role. In relation with experimental results we review models of basic molecular and cellular mechanisms involved in the signal transduction from the myocyte stimulation to the activation of the contractile apparatus. We concentrate on mechanisms for encoding of input signals into Ca2+ signals and the mechanisms for their decoding. The mechanisms are arranged into a general scheme of cellular signaling, the so-called bow-tie architecture of signaling, in which calcium plays the role of a common media for cellular signals and links the encoding and decoding part. The encoding of calcium signals in airway myocytes is better known and is presented in more detail. Inparticular, we focus on three recent models taking into account the intracellular calcium handling and ion fluxes through the plasma membrane. Themodel of membrane conductances was originally proposed for predicting membrane depolarization and voltage-dependent Ca2+ influx triggered by initialcytosolic Ca2+ increase as observed on cholinergic stimulation. Cellular models of intracellular Ca2+ handling were developed to investigate the role of a mixed population of InsP3 receptor isoforms and the cellular environment in the occurrence of Ca2+ oscillations, and the respective role ofthe sarcoplasmic reticulum, mitochondria, and cytosolic Ca2+-binding proteins in cytosolic Ca2+ clearance. Modeling the mechanisms responsible for the decoding of calcium signals is developed in a lesser extent; however, the most recent theoretical studies are briefly presented in relation with the known experimental results.
Keywords: biophysics, mathematical modelling, modelling, calcium oscillations, contractions, airway smooth muscle cells, muscle cells, smooth muscles, encoding, decoding, bow-tie structures
Published: 07.06.2012; Views: 1147; Downloads: 17
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Bacterial chemotaxis and entropy production
Paško Županović, Milan Brumen, Marko Jagodič, Davor Juretić, 2010, original scientific article

Abstract: Entropy production is calculated for bacterial chemotaxis in the case of a migrating band of bacteria in a capillary tube. It is found that the speed of the migrating band is a decreasing function of the starting concentration of the metabolizable attractant. The experimentally found dependence of speed on the starting concentration of galactose, glucose and oxygen is fitted with power-law functions. It is found that the corresponding exponents lie within the theoretically predicted interval. The effect of the reproduction of bacteria on band speed is considered, too. The acceleration of the band is predicted due to the reproduction rate of bacteria. The relationship between chemotaxis, the maximum entropy production principle and the formation of self-organizing structure is discussed.
Keywords: biofizika, bakterijska kemotaksa, hitrost reprodukcije, princip maksimalne produkcije entropije, biophysics, bacterial chemotaxis, reproduction rate, princip maksimalne produkcije entropije
Published: 10.07.2015; Views: 480; Downloads: 65
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10.
Progressive glucose stimulation of islet beta cells reveals a transition from segregated to integrated modular functional connectivity patterns
Rene Markovič, Andraž Stožer, Marko Gosak, Jurij Dolenšek, Marko Marhl, Marjan Rupnik, 2015, original scientific article

Abstract: Collective beta cell activity in islets of Langerhans is critical for the supply of insulin within an organism. Even though individual beta cells are intrinsically heterogeneous, the presence of intercellular coupling mechanisms ensures coordinated activity and a well-regulated exocytosis of insulin. In order to get a detailed insight into the functional organization of the syncytium, we applied advanced analytical tools from the realm of complex network theory to uncover the functional connectivity pattern among cells composing the intact islet. The procedure is based on the determination of correlations between long temporal traces obtained from confocal functional multicellular calcium imaging of beta cells stimulated in a stepwise manner with a range of physiological glucose concentrations. Our results revealed that the extracted connectivity networks are sparse for low glucose concentrations, whereas for higher stimulatory levels they become more densely connected. Most importantly, for all ranges of glucose concentration beta cells within the islets form locally clustered functional sub-compartments, thereby indicating that their collective activity profiles exhibit a modular nature. Moreover, we show that the observed non-linear functional relationship between different network metrics and glucose concentration represents a well-balanced setup that parallels physiological insulin release.
Keywords: endocrinology, computational biophysics, calcium signalling, biological physics
Published: 23.06.2017; Views: 557; Downloads: 203
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