Anomalous Reactances in Electrodiffusion Systems

A frequency response analysis of a constrained diffusion boundary has been made by linearizing the Nernst-Planck equations for a small applied AC current. The number of time constants and their dependence on ionic concentrations and electric field as well as membrane parameters such as dielectric constant, thickness, etc. have been evaluated by this method. Numerical solutions have been carried out for cases when the Planck charging time can be neglected and the results are presented in the form of impedance loci. These impedance loci show that if the membrane separates two univalent electrolytes with a common anion it will exhibit a combined capacitative inductive response with a 90° phase angle. The dependence of these anomalous reactances on ionic concentrations and the electric field is consistent with the behavior of the Hodgkin-Huxley axon suggesting that a homogeneous electrodiffusion regime could be adequate as a basic model for the kinetic behavior of biological membranes.     

Unilamellar-Vesicle Systems as Model for Studying Membrane Permeabilization by Polyene Antibiotics

Abstract

Permeabilization of phospholipid/sterol unilamellar vesicles by polyene antibiotics (amphotericin B and lucensomycin) was studied by measuring proton leakage with a pH-stat method. The percentage of proton release was directly related to the antibiotic concentration. Using ergosterol-containing vesicles, a relevant proton efflux was induced by micromolar concentrations of amphotericin B, whereas lucensomycin caused membrane permeabilization at higher concentrations (0.1 mM). Cholesterol-containing vesicles were less sensible to the lytic action of polyenes. When amphotericin B was carried in cholesterol-containing liposomes, the selectivity towards ergosterol-containing vesicles was enhanced. An increase in drug selectivity was also observed by dissolving amphotericin B in fresh human plasma. At concentrations one order of magnitude lower than those necessary to induce a detectable proton efflux, lucensomycin seemed to protect the vesicles from the subsequent permeabilizing action of amphotericin B.     

Nitric Oxide Treatment for the Control of Reverse Osmosis Membrane Biofouling

Biofouling remains a key challenge for membrane-based water treatment systems. This study investigated the dispersal potential of the nitric oxide (NO) donor compound, PROLI NONOate, on single- and mixed-species biofilms formed by bacteria isolated from industrial membrane bioreactor and reverse osmosis (RO) membranes. The potential of PROLI NONOate to control RO membrane biofouling was also examined. Confocal microscopy revealed that PROLI NONOate exposure induced biofilm dispersal in all but two of the bacteria tested and successfully dispersed mixed-species biofilms. The addition of 40 μM PROLI NONOate at 24-h intervals to a laboratory-scale RO system led to a 92% reduction in the rate of biofouling (pressure rise over a given period) by a bacterial community cultured from an industrial RO membrane. Confocal microscopy and extracellular polymeric substances (EPS) extraction revealed that PROLI NONOate treatment led to a 48% reduction in polysaccharides, a 66% reduction in proteins, and a 29% reduction in microbial cells compared to the untreated control. A reduction in biofilm surface coverage (59% compared to 98%, treated compared to control) and average thickness (20 μm compared to 26 μm, treated compared to control) was also observed. The addition of PROLI NONOate led to a 22% increase in the time required for the RO module to reach its maximum transmembrane pressure (TMP), further indicating that NO treatment delayed fouling. Pyrosequencing analysis revealed that the NO treatment did not significantly alter the microbial community composition of the membrane biofilm. These results present strong evidence for the application of PROLI NONOate for prevention of RO biofouling.     

How does Amphotericin B Work?: Studies on Model Membrane Systems

Abstract

The drug Amphotericin B is a very important antifungal agent as well as one of the first model systems for transmembrane pore structures. The most widely accepted model for the anticellular activity of this drug involves the formation of 1:1 Amphotericin/ sterol aggregates which subsequently associate into a transmembrane barrel with a large -OH lined aqueous pore down the middle. The stronger association of Amphotericin with ergosterol versus cholesterol explains the higher toxicity toward fungi. However, conflicting membrane permeability data concerning Amphotericin channel ion selectivity, sterol requirements, and mode of delivery has accumulated over the past fifteen years and suggests there exists a multiplicity of AmB channel structures and modes of action. Some of these mechanisms of action may be even more relevant clinically than the Amphotericin/sterol pore structure. Some of the anticellular membrane damage caused by Amphotericin may be due to formation of membrane defects and non-bilayer phases, channels without sterol or even induction of oxidative damage. In this article we present a survey of recent observations on AmB's activity on model membrane systems. As such, we are mostly concerned with liposome and planar bilayer studies. Some of the newer models explaining AmB s differential effects on cholesterol versus ergosterol containing membranes are presented along with a brief overview of membrane disruption models based on current research on membrane-active amphiphilic peptides. A synthesis and reconciliation of many of these diverse observations is attempted in a model which can accommodate most aspects of the classical sterol/Amphotericin barrel model and more recent observations as well.     

Analysis of Reaction-Diffusion Systems with Anomalous Subdiffusion

Reaction-diffusion equations are the cornerstone of modeling biochemical systems with spatial gradients, which are relevant to biological processes such as signal transduction. Implicit in the formulation of these equations is the assumption of Fick's law, which states that the local diffusive flux of species i is proportional to its concentration gradient; however, in the context of complex fluids such as cytoplasm and cell membranes, the use of Fick's law is based on empiricism, whereas evidence has been mounting that such media foster anomalous subdiffusion (with mean-squared displacement increasing less than linearly with time) over certain length scales. Particularly when modeling diffusion-controlled reactions and other systems where the spatial domain is considered semi-infinite, assuming Fickian diffusion might not be appropriate. In this article, two simple, conceptually extreme models of anomalous subdiffusion are used in the framework of Green's functions to demonstrate the solution of four reaction-diffusion problems that are well known in the biophysical context of signal transduction: fluorescence recovery after photobleaching, the Smolochowski limit for diffusion-controlled reactions in solution, the spatial range of a diffusing molecule with finite lifetime, and the collision coupling mechanism of diffusion-controlled reactions in two dimensions. In each case, there are only subtle differences between the two subdiffusion models, suggesting how measurements of mean-squared displacement versus time might generally inform models of reactive systems with partial diffusion control.     

Anomalous Diffusion Induced by Cristae Geometry in the Inner Mitochondrial Membrane

Diffusion of inner membrane proteins is a prerequisite for correct functionality of mitochondria. The complicated structure of tubular, vesicular or flat cristae and their small connections to the inner boundary membrane impose constraints on the mobility of proteins making their diffusion a very complicated process. Therefore we investigate the molecular transport along the main mitochondrial axis using highly accurate computational methods. Diffusion is modeled on a curvilinear surface reproducing the shape of mitochondrial inner membrane (IM). Monte Carlo simulations are carried out for topologies resembling both tubular and lamellar cristae, for a range of physiologically viable crista sizes and densities. Geometrical confinement induces up to several-fold reduction in apparent mobility. IM surface curvature per se generates transient anomalous diffusion (TAD), while finite and stable values of projected diffusion coefficients are recovered in a quasi-normal regime for short- and long-time limits. In both these cases, a simple area-scaling law is found sufficient to explain limiting diffusion coefficients for permeable cristae junctions, while asymmetric reduction of the junction permeability leads to strong but predictable variations in molecular motion rate. A geometry-based model is given as an illustration for the time-dependence of diffusivity when IM has tubular topology. Implications for experimental observations of diffusion along mitochondria using methods of optical microscopy are drawn out: a non-homogenous power law is proposed as a suitable approach to TAD. The data demonstrate that if not taken into account appropriately, geometrical effects lead to significant misinterpretation of molecular mobility measurements in cellular curvilinear membranes.     

Membrane Systems of Crab Fibers

An electron microscopic study of internal and surface-connectedmembrane systems of leg muscle of the crab shows that thereare three kinds of surface-connected membrane systems in additionto an intracellular sarcoplasmic reticulum (SR). One is a systemof large infoldings of the sarcolemma referred to as clefts.These are longitudinally-oriented, flattened infoldings of boththe plasma membrane and the fibrous sheath of the fiber, andwere probably seen earlier with the light microscope. Extendinginto the fiber both from these clefts and from the free fibersurface are two systems of tubules of much smaller caliber,the Z tubules and the A tubules. The Z tubules are located,as their name indicates, near the Z lines of the myofibrils,and are thought to be attached to them mechanically. The A tubulesare found in pairs, near the ends of each A band, and are closelybound to the SR in two-part structures called dyads. Local-activationexperiments, like those done earlier by Huxley and Taylor, suggestthat the A tubules are involved in excitation-contraction coupling;no such experimental suggestion of function exists for the Ztubules.     

Affinity of Alkylphosphocholines to Biological Membrane of Prostate Cancer: Studies in Natural and Model Systems

The effectiveness of two alkylphosphocholines (APCs), hexadecylphosphocholine (miltefosine) and erucylphosphocholine to combat prostate cancer has been studied in vitro with artificial cancerous membrane, modelled with the Langmuir monolayer technique, and on cell line (Du-145). Studies performed with the Langmuir method indicate that both the investigated drugs have the affinity to the monolayer mimicking prostate cancer membrane (composed of cholesterol:POPC = 0.428) and the drug-membrane interactions are stronger for erucylphosphocholine as compared to hexadecylphosphocholine. Moreover, both studied drugs were found to fluidize the model membrane, which may lead to apoptosis. Indeed, biological studies confirmed that in Du-145 cell line both investigated alkylphosphocholines cause cell death primarily by apoptosis while necrotic cells constitute only a small percentage of APC-treated cells.     

Membrane Polypeptides associated with Photochemical Systems

WE wish to report a specific relationship between certain chloroplast membrane polypeptides and functional properties of the chloroplast usually associated with one or the other of the two photochemical systems (designated PSI and PSII). We have known this by the analysis of the chloroplast membrane polypeptides of the wild type strain of the unicellular green alga Chlamydomonas reinhardi and of mutant strains derived from it which have lost the capacity to carry out normal photosynthesis. These mutants have been characterized by the loss of particular membrane-bound components of the photosynthetic electron transport chain^1,2. This relationship is supported by analysis of the membrane polypeptides obtained from chloroplast fractions of wild type C. reinhardi and spinach enriched for reactions characteristic of either PSI or PSII by fractionation of the membranes either with digitonin^3,4 or ‘Triton X-100’⁵.     

物质跨膜转运的通道转运

简单列举物质跨膜运输的几种方式,详细介绍通道转运中4种重要的通道:电位门通道、配体门通道、环核苷酸门通道和机械门通道及水通道概念的提出。总结通道易化扩散的特点。     

Minimalist Model Systems Reveal Similarities and Differences between Membrane Interaction Modes of MCL1 and BAK

Proteins belonging to the BCL2 family are key modulators of apoptosis that establish a complex network of interactions among themselves and with other cellular factors to regulate cell fate. It is well established that mitochondrial membranes are the main locus of action of all BCL2 family proteins, but it is difficult to obtain a precise view of how BCL2 family members operate at the native mitochondrial membrane environment during apoptosis. Here, we used minimalist model systems and multiple fluorescence-based techniques to examine selected membrane activities of MCL1 and BAK under apoptotic-like conditions. We show that three distinct apoptosis-related factors (i.e. the BCL2 homology 3 ligand cBID, the mitochondrion-specific lipid cardiolipin, and membrane geometrical curvature) all promote membrane association of BCL2-like structural folds belonging to both MCL1 and BAK. However, at the same time, the two proteins exhibited distinguishing features in their membrane association modes under apoptotic-like conditions. In addition, scanning fluorescence cross-correlation spectroscopy and FRET measurements revealed that the BCL2-like structural fold of MCL1, but not that of BAK, forms stable heterodimeric complexes with cBID in a manner adjustable by membrane cardiolipin content and curvature degree. Our results add significantly to a growing body of evidence indicating that the mitochondrial membrane environment plays a complex and active role in the mode of action of BCL2 family proteins.     

Protein Secretion and Membrane Insertion Systems in Gram-Negative Bacteria

In contrast to other organisms, gram-negative bacteria have evolved numerous systems for protein export. Eight types are known that mediate export across or insertion into the cytoplasmic membrane, while eight specifically mediate export across or insertion into the outer membrane. Three of the former secretory pathway (SP) systems, type I SP (ISP, ABC), IIISP (Fla/Path) and IVSP (Conj/Vir), can export proteins across both membranes in a single energy-coupled step. A fourth generalized mechanism for exporting proteins across the two-membrane envelope in two distinct steps (which we here refer to as type II secretory pathways [IISP]) utilizes either the general secretory pathway (GSP or Sec) or the twin-arginine targeting translocase for translocation across the inner membrane, and either the main terminal branch or one of several protein-specific export systems for translocation across the outer membrane. We here survey the various well-characterized protein translocation systems found in living organisms and then focus on the systems present in gram-negative bacteria. Comparisons between these systems suggest specific biogenic, mechanistic and evolutionary similarities as well as major differences.     

Osmosis in the balance

A simple investigation into the changes in mass of plant tissue when placed in solutions of different osmotic concentration is described. It is suitable for pupils at early secondary or even late primary levels. The merits of the investigation include its low cost, the easily availability of the materials required, and the ease with which the reversibility of the process can be demonstrated. It is good test of the manipulative skills of students and raises important questions as to the cause(s) of the changes in mass of the plant tissue.     

Model Hydrophobic Ion Exchange Membrane

Two models of hydrophobic ion exchange membranes were examined theoretically with regard to the characteristics of cellulose acetate-nitrate membranes saturated with hydrophobic solvents. The first model, consisting of fixed negative sites dispersed in a homogeneous medium of low dielectric constant, was shown to be invalid for the experimental membranes. The second model, consisting of fixed negative sites in an aqueous channel surrounded by a medium of low dielectric constant, explains many properties of the cellulose acetate-nitrate hydrophobic membranes and was analyzed in some detail. Organic cations can enter the membranes through the hydrophobic phase as well as through the aqueous channels. The mechanism of counterion movement in such a model is assumed to consist of exchange of vacancies and or double-occupied sites positions. The presence of the medium of low dielectric constant around the aqueous channel increases the “self”-energy of the ions in the channel and the electrostatic interaction between a fixed site and a counterion in the membrane. Both these factors can account for the marked dependence of ion mobility in the aqueous channels on the dielectric constant of the surrounding medium. The model predicts membrane preference for monovalent counterions over divalent ones.     

An Ion Displacement Membrane Model

The usual assumption in treating the diffusion of ions in an electric field has been that the movement of each ion is independent of the movement of the others. The resulting equation for diffusion by a succession of spontaneous jumps has been well stated by Parlin and Eyring. This paper will consider one simple case in which a different assumption is reasonable. Diffusion of monovalent positive ions is considered as a series of jumps from one fixed negative site to another. The sites are assumed to be full (electrical neutrality). Interaction occurs by the displacement of one ion by another. An ion leaves a site if and only if another ion, not necessarily of the same species, attempts to occupy the same site. Flux ratios and net fluxes are given as functions of the electrical potential, concentration ratios, and number of sites encountered in crossing the membrane. Quantitative comparisons with observations of Hodgkin and Keynes are presented.     

Mathematical Model for Rhythmic Protoplasmic Movement in the True Slime Mold

The plasmodium of the true slime mold Physarum polycephalum is a large amoeboid organism that displays “smart” behavior such as chemotaxis and the ability to solve mazes and geometrical puzzles. These amoeboid behaviors are based on the dynamics of the viscoelastic protoplasm and its biochemical rhythms. By incorporating both these aspects, we constructed a mathematical model for the dynamics of the organism as a first step towards understanding the relation between protoplasmic movement and its unusual abilities. We tested the validity of the model by comparing it with physiological observation. Our model reproduces fundamental characteristics of the spatio-temporal pattern of the rhythmic movement: (1) the antiphase oscillation between frontal tip and rear when the front is freely extending; (2) the asynchronous oscillation pattern when the front is not freely extending; and (3) the formation of protoplasmic mounds over a longer time scale. Both our model and physiological observation suggest that cell stiffness plays a primary role in plasmodial behaviors, in contrast to the conventional theory of coupled oscillator systems.     

Role of Vacuolar Membrane Transport Systems in Plant Salinity Tolerance

About 20% of all irrigated land is adversely affected by salinity hazards and therefore understanding plant defense mechanisms against salinity will have great impact on plant productivity. In the last decades, comprehension of salinity resistance at molecular level has been achieved through the identification of key genes encoding biomarker proteins underpinning salinity tolerance. Implication of the vacuolar transport systems in plant salinity tolerance is one example of these central mechanisms rendering tolerance to saline stress. One important organelle in plant cells is the central vacuole that plays pivotal multiple roles in cell functioning under normal and stress conditions. This review thus attempts to address different lines of evidence supporting the role of the vacuolar membrane transport systems in plant salinity tolerance. Vacuolar transport systems include Na⁺(K⁺)/H⁺ antiporters, V-ATPase, V-PPase, Ca²⁺/H⁺ exchangers, Ca²⁺-ATPase, ion channels, aquaporins, and ABC transporters. They contribute essentially in retaining a high cytosolic K⁺/Na⁺ ratio, K⁺ level, sequestrating Na⁺ and Cl^? into vacuoles, as well as regulation of other salinity responsive pathways. However, little is known about the regulation and functions of some of the vacuolar transporters under salinity stress and therefore need more exploration and focus. Numerous studies demonstrated that the activities of the vacuolar transporters are upregulated in response to salinity stress, confirming their central roles in salinity tolerance mechanism. The second line of evidence is that manipulation of one of the genes encoding the vacuolar transport proteins results in some successful improvement of plant salinity tolerance. Therefore, transgene pyramiding of more than one gene for developing genotypes with better and strong salinity tolerance and productivity should gain more attention in future research. In addition, we should move step further and verify the experimental data obtained from either a greenhouse or controlled environment into field trials in order to support our claims.