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Sample records for lipid layer determined

  1. Composite S-layer lipid structures

    PubMed Central

    Schuster, Bernhard; Sleytr, Uwe B.

    2010-01-01

    Designing and utilization of biomimetic membrane systems generated by bottom-up processes is a rapidly growing scientific and engineering field. Elucidation of the supramolecular construction principle of archaeal cell envelopes composed of S-layer stabilized lipid membranes led to new strategies for generating highly stable functional lipid membranes at meso- and macroscopic scale. In this review, we provide a state of the art survey how S-layer proteins, lipids, and polysaccharides may be used as basic building blocks for the assembly of S-layer supported lipid membranes. These biomimetic membrane systems are distinguished by a nanopatterned fluidity, enhanced stability and longevity and thus, provide a dedicated reconstitution matrix for membrane-active peptides and transmembrane proteins. Exciting areas for application of composite S-layer membrane systems concern sensor systems involving specific membrane functions. PMID:19303933

  2. Keratin-lipid structural organization in the corneous layer of snake.

    PubMed

    Ripamonti, Alberto; Alibardi, Lorenzo; Falini, Giuseppe; Fermani, Simona; Gazzano, Massimo

    2009-12-01

    The shed epidermis (molt) of snakes comprises four distinct layers. The upper two layers, here considered as beta-layer, contain essentially beta-keratin. The following layer, known as mesos-layer, is similar to the human stratum corneum, and is formed by thin cells surrounded by intercellular lipids. The latter layer mainly contains alpha-keratin. In this study, the molecular assemblies of proteins and lipids contained in these layers have been analyzed in the scale of two species of snakes, the elapid Tiger snake (TS, Notechis scutatus) and the viperid Gabon viper (GV, Bitis gabonica). Scanning X-ray micro-diffraction, FTIR and Raman spectroscopies, thermal analysis, and scanning electron microscopy experiments confirm the presence of the three layers in the GV skin scale. Conversely, in the TS molt a typical alpha-keratin layer appears to be absent. In the latter, experimental data suggest the presence of two domains similar to those found in the lipid intercellular matrix of stratum corneum. X-ray diffraction data also allow to determine the relative orientation of keratins and lipids. The keratin fibrils are randomly oriented inside the layers parallel to the surface of scales while the lipids are organized in lamellar structures having aliphatic chains normal to the scale surface. The high ordered lipid organization in the mature mesos layer probably increases its effectiveness in limiting water-loss. PMID:19280640

  3. Determination of phosphorus in cereal lipids.

    PubMed

    Kovacs, M I

    1986-05-01

    The effect of digestion methods on the determination of phosphorus in cereal lipids was reinvestigated. Samples were either digested with sulfuric acid or ashed in a muffle furnace at 600 degrees C. The standard deviation and the coefficient of variation were significantly higher for the acid-digested samples. Ashing gave more reliable results, especially when large amounts of lipid material had to be oxidized. PMID:3728960

  4. Role of neutral lipids in tear fluid lipid layer: coarse-grained simulation study.

    PubMed

    Telenius, Jelena; Koivuniemi, Artturi; Kulovesi, Pipsa; Holopainen, Juha M; Vattulainen, Ilpo

    2012-12-11

    Tear fluid lipid layer (TFLL) residing at the air-water interface of tears has been recognized to play an important role in the development of dry eye syndrome. Yet, the composition, structure, and mechanical properties of TFLL are only partly known. Here, we report results of coarse-grained simulations of a lipid layer comprising phospholipids, free fatty acids, cholesteryl esters, and triglycerides at the air-water interface to shed light on the properties of TFLL. We consider structural as well as dynamical properties of the lipid layer as a function of surface pressure. Simulations revealed that neutral lipids reside heterogeneously between phospholipids at relatively low pressures but form a separate hydrophobic phase with increasing surface pressure, transforming the initial lipid monolayer to a two-layered structure. When the model of TFLL was compared to a one-component phospholipid monolayer system, we found drastic differences in both structural and dynamical properties that explain the prominent role of neutral lipids as stabilizers of the TFLL. Based on our results, we suggest that neutral lipids are able to increase the stability of the TFLL by modulating its dynamical and structural behavior, which is important for the proper function of tear film. PMID:23151187

  5. Combined urea-thin layer chromatography and silver nitrate-thin layer chromatography for micro separation and determination of hard-to-detect branched chain fatty acids in natural lipids.

    PubMed

    Yan, Yuanyuan; Wang, Xingguo; Liu, Yijun; Xiang, Jingying; Wang, Xiaosan; Zhang, Huijun; Yao, Yunping; Liu, Ruijie; Zou, Xiaoqiang; Huang, Jianhua; Jin, Qingzhe

    2015-12-18

    A simple, fast and efficient procedure was developed for micro separation and enrichment of branched chain fatty acids (BCFA) from natural products using successive thin layer chromatography (TLC) technique coupling novel urea-TLC with AgNO3-TLC, which rely on the formation of urea adduction and AgNO3 bonding in methanol. These natural lipids contain a significant amount of straight chain fatty acids (FA). Fresh and fast urea-TLC and AgNO3-TLC plate making techniques were developed with more even coating and less coating material contamination before being utilized for separation. Goat milk fat was used as a model. Various experimental parameters that affect urea-TLC and AgNO3-TLC separation of BCFA were investigated and optimized, including coating of urea, concentration of original oil sample, mobile phase and sample application format. High efficiency of removal of straight chain FA was achieved with a low amount of sample in an easy and fast way. A total BCFA mix with much higher purity than previous studies was successfully achieved. The developed method has also been applied for the concentration and analysis of BCFA in cow milk fat and Anchovy oil. PMID:26614174

  6. Combined urea-thin layer chromatography and silver nitrate-thin layer chromatography for micro separation and determination of hard-to-detect branched chain fatty acids in natural lipids.

    PubMed

    Yan, Yuanyuan; Wang, Xingguo; Liu, Yijun; Xiang, Jingying; Wang, Xiaosan; Zhang, Huijun; Yao, Yunping; Liu, Ruijie; Zou, Xiaoqiang; Huang, Jianhua; Jin, Qingzhe

    2015-12-18

    A simple, fast and efficient procedure was developed for micro separation and enrichment of branched chain fatty acids (BCFA) from natural products using successive thin layer chromatography (TLC) technique coupling novel urea-TLC with AgNO3-TLC, which rely on the formation of urea adduction and AgNO3 bonding in methanol. These natural lipids contain a significant amount of straight chain fatty acids (FA). Fresh and fast urea-TLC and AgNO3-TLC plate making techniques were developed with more even coating and less coating material contamination before being utilized for separation. Goat milk fat was used as a model. Various experimental parameters that affect urea-TLC and AgNO3-TLC separation of BCFA were investigated and optimized, including coating of urea, concentration of original oil sample, mobile phase and sample application format. High efficiency of removal of straight chain FA was achieved with a low amount of sample in an easy and fast way. A total BCFA mix with much higher purity than previous studies was successfully achieved. The developed method has also been applied for the concentration and analysis of BCFA in cow milk fat and Anchovy oil.

  7. The electroanalytical approach to lipid peroxide determinations.

    PubMed

    Funk, M O

    1987-01-01

    An electroanalytical method for the determination of lipid peroxides in physiological material is described. The technique is based on electrochemical detection for HPLC as the means for enhancing sensitivity. Samples containing organic peroxides, including lipid peroxides, can be analyzed directly using a modified polarographic detector (Lloyd, J.B.F.; Optimization of the operational parameters of the supported mercury drop electrode detector in high performance liquid chromatography. Anal. Chim. Acta 154:121-131; 1983.) for reversed phase HPLC determinations. Detection limits for fatty acid hydroperoxides were found to be in the low nanogram range.

  8. Lipid Layers on Polyelectrolyte Multilayers: Understanding Lipid-Polyelectrolyte Interactions and Applications on the Surface Engineering of Nanomaterials.

    PubMed

    Diamanti, Eleftheria; Gregurec, Danijela; Gabriela, Romero; Cuellar, J L; Donath, E; Moya, S E

    2016-06-01

    In this manuscript we review work of our group on the assembly of lipid layers on top of polyelectrolyte multilayers (PEMs). The assembly of lipid layers with zwitterionic and charged lipids on PEMs is studied as a function of lipid and polyelectrolyte composition by the Quartz Crystal Microbalance. Polyelectrolyte lipid interactions are studied by means of Atomic Force Spectroscopy. We also show the coating of lipid layers for engineering different nanomaterials, i.e., carbon nanotubes and poly(lactic-co-glycolic) nanoparticles and how these can be used to decrease in vitro toxicity and to direct the intracellular localization of nanomaterials.

  9. Lipid Layers on Polyelectrolyte Multilayers: Understanding Lipid-Polyelectrolyte Interactions and Applications on the Surface Engineering of Nanomaterials.

    PubMed

    Diamanti, Eleftheria; Gregurec, Danijela; Gabriela, Romero; Cuellar, J L; Donath, E; Moya, S E

    2016-06-01

    In this manuscript we review work of our group on the assembly of lipid layers on top of polyelectrolyte multilayers (PEMs). The assembly of lipid layers with zwitterionic and charged lipids on PEMs is studied as a function of lipid and polyelectrolyte composition by the Quartz Crystal Microbalance. Polyelectrolyte lipid interactions are studied by means of Atomic Force Spectroscopy. We also show the coating of lipid layers for engineering different nanomaterials, i.e., carbon nanotubes and poly(lactic-co-glycolic) nanoparticles and how these can be used to decrease in vitro toxicity and to direct the intracellular localization of nanomaterials. PMID:27427617

  10. Effect of meibomian lipid layer on evaporation of tears

    NASA Astrophysics Data System (ADS)

    Miano, F.; Calcara, M.; Giuliano, F.; Millar, T. J.; Enea, V.

    2004-07-01

    The outer interface of a tear film was studied with the aid of a model system able to investigate the interfacial phenomena derived from the spreading of an insoluble lipid multilayer onto a tear-like aqueous fluid. The interactions of such a layer with proteins dissolved in the aqueous phase beneath were also investigated. Emphasis was given to evaporation phenomena because the increased rate of tear evaporation in humans is often related to a number of ocular dysfunctions. The model tear was studied as a pendant drop that permitted a functional evaluation of the effect of lipids and proteins upon the evaporation of water from the tear film.

  11. Tear film lipid layer: A molecular level view.

    PubMed

    Cwiklik, Lukasz

    2016-10-01

    Human cornea is covered by an aqueous tear film, and the outermost layer of the tear film is coated by lipids. This so-called tear film lipid layer (TFLL) reduces surface tension of the tear film and helps with the film re-spreading after blinks. Alterations of tear lipids composition and properties are related to dry eye syndrome. Therefore, unveiling structural and functional properties of TFLL is necessary for understanding tear film function under both normal and pathological conditions. Key properties of TFLL, such as resistance against high lateral pressures and ability to spread at the tear film surface, are directly related to the chemical identity of TFLL lipids. Hence, a molecular-level description is required to get better insight into TFLL properties. Molecular dynamics simulations are particularly well suited for this task and they were recently used for investigating TFLL. The present review discusses molecular level organization and properties of TFLL as seen by these simulation studies. This article is part of a Special Issue entitled: Biosimulations edited by Ilpo Vattulainen and Tomasz Róg.

  12. [Abnormal of tear lipid layer and recent advances in clinical study of dry eye].

    PubMed

    Xiao, Xin-Ye; Liu, Zu-Guo

    2012-03-01

    Dry eye is a common disease in the ophthalmological clinic, which is related to the dysfunction of tear film. The tear film is composed of lipid layer, aqueous layer and mucin layer (or lipid layer, aqueous/mucin layer). The lipid of the outmost layer derived from Meibomian gland and distributed on the tear film after blinking can decrease the evaporation and stabilize the tear film. The thickness, quality, and distribution of lipid layer are impaired in many dry eye patients, hence restoring the physiological function of lipid layer may be crucial for the treatment of this kind of dry eye. The lipid artificial tears manifest great effects on increasing lipid layer thickness, stabilizing tear film, improving Meibomian gland dysfunction, and promoting tear film distribution.

  13. Lipid and protein maps defining arterial layers in atherosclerotic aorta.

    PubMed

    Martin-Lorenzo, Marta; Balluff, Benjamin; Maroto, Aroa S; Carreira, Ricardo J; van Zeijl, Rene J M; Gonzalez-Calero, Laura; de la Cuesta, Fernando; Barderas, Maria G; Lopez-Almodovar, Luis F; Padial, Luis R; McDonnell, Liam A; Vivanco, Fernando; Alvarez-Llamas, Gloria

    2015-09-01

    Subclinical atherosclerosis cannot be predicted and novel therapeutic targets are needed. The molecular anatomy of healthy and atherosclerotic tissue is pursued to identify ongoing molecular changes in atherosclerosis development. Mass Spectrometry Imaging (MSI) accounts with the unique advantage of analyzing proteins and metabolites (lipids) while preserving their original localization; thus two dimensional maps can be obtained. Main molecular alterations were investigated in a rabbit model in response to early development of atherosclerosis. Aortic arterial layers (intima and media) and calcified regions were investigated in detail by MALDI-MSI and proteins and lipids specifically defining those areas of interest were identified. These data further complement main findings previously published in J Proteomics (M. Martin-Lorenzo et al., J. Proteomics. (In press); M. Martin-Lorenzo et al., J. Proteomics 108 (2014) 465-468.) [1,2]. PMID:26217810

  14. Profiling the Triacylglyceride Contents in Bat Integumentary Lipids by Preparative Thin Layer Chromatography and MALDI-TOF Mass Spectrometry

    PubMed Central

    Pannkuk, Evan L.; Risch, Thomas S.; Savary, Brett J.

    2013-01-01

    The mammalian integument includes sebaceous glands that secrete an oily material onto the skin surface. Sebum production is part of the innate immune system that is protective against pathogenic microbes. Abnormal sebum production and chemical composition are also a clinical symptom of specific skin diseases. Sebum contains a complex mixture of lipids, including triacylglycerides, which is species-specific. The broad chemical properties exhibited by diverse lipid classes hinder the specific determination of sebum composition. Analytical techniques for lipids typically require chemical derivatizations that are labor-intensive and increase sample preparation costs. This paper describes how to extract lipids from mammalian integument, separate broad lipid classes by thin-layer chromatography, and profile the triacylglyceride contents using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This robust method enables a direct determination of the triacylglyceride profiles among species and individuals, and it can be readily applied to any taxonomic group of mammals. PMID:24056580

  15. Stability and phase separation in mixed monopolar lipid/bolalipid layers.

    PubMed

    Longo, Gabriel S; Thompson, David H; Szleifer, I

    2007-10-15

    The phase stability of a fluid lipid layer that is a mixture of conventional monopolar lipids and C20 bipolar bolalipids was studied using a mean field theory that explicitly includes molecular details and configurational properties of the lipid molecules. The effect of changing the fraction of bolalipids, as well as the length of the hydrocarbon chain of the monopolar lipids, was probed. A phase separation between two liquid lipid phases was found when a mismatch exists in the optimal hydrophobic thicknesses of the pure bolalipid and monopolar lipid layers. The lipid mixture phase separates into a thin bolalipid-rich layer and a thicker monopolar-rich layer. The thin membrane phase is mainly composed of transmembrane bolalipid molecules whose polar heads are positioned at opposite membrane-water interfaces. In the monopolar lipid-rich phase, bolalipids are the minor component and most of them assume a looping configuration where both headgroups are present at the same membrane-water interface. For mixed layers that form a single lipid phase across all bolalipid concentrations, the hairpin-transmembrane ratio strongly depends on the hydrocarbon chain length of the monopolar lipid and the bolalipid concentration. The C-D bond order parameters of the different species have been calculated. Our findings suggest that the concentration-dependent phase transition should be experimentally observable by measuring of the order parameters through quadrupolar splitting experiments. The driving force for the phase separation in the monopolar lipid/bolalipid mixture is the packing mismatch between hydrophobic regions of the monopolar lipid hydrocarbon chains and the membrane-spanning bolalipid chains. The results from the molecular theory may be useful in the design of stable lipid layers for integral membrane protein sensing.

  16. Tear lipid layer deficiency associated with incomplete blinking: A case report

    PubMed Central

    2013-01-01

    Background Meibomian gland obstruction induces hyposecretion of tear film lipids, which results in lipid layer deficiency and evaporative dry eye. Unfortunately, the importance of blinking in meibomian gland dysfunction has been largely overlooked, and it is not known whether incomplete blinking causes tear lipid deficiency, even in the unobstructed meibomian glands. Case presentation A 38-year-old woman suffering from foreign body sensations in her eyes was examined. The cornea was clear and tear secretion was normal. Lid margin abnormalities were not observed and the meibum was clear. However, the lipid layer was very thin, and the patient was given a diagnosis of incomplete blinking. The patient was made aware of her condition and asked to blink consciously and completely. After that, an immediate increase in lipid flow was observed. Conclusion Tear lipid layer deficiency can occur with incomplete blinking, even though meibomian gland structures are intact. This case highlights the importance of complete blinking. PMID:23855887

  17. Debye-Hückel theory of mixed charged-zwitterionic lipid layers

    NASA Astrophysics Data System (ADS)

    Mengistu, D. H.; May, S.

    2008-07-01

    Modeling electrostatic properties of macroions such as charged lipid membranes is especially simple within linear Debye-Hückel theory where analytical solutions are often available. Charged lipid layers typically also contain zwitterionic lipids that possess a large headgroup dipole. We incorporate the presence of zwitterionic lipids into the Debye-Hückel description and derive analytical expressions for the free energies of isolated and interacting lipid layers. Our approach accounts for two major characteristic features of zwitterionic lipids, the firm linkage of the dipole's negatively charged phosphate end to the lipid chain and the comparatively large conformational freedom of the opposite, positively charged, dipole end. This leads to differences in structural properties of mixed anionic and cationic lipid layers that are known from experiment and that the Debye-Hückel description qualitatively recovers. Most notably, this includes the different dependencies of the average cross-sectional area per lipid on the composition of mixed anionic and cationic membranes. In addition, we study the predictions of Debye-Hückel theory regarding the electrostatic influence of zwitterionic lipids on the stability of non-ideally mixed membranes as well as on the adsorption energy of oppositely charged macroions.

  18. Protein Crowding Is a Determinant of Lipid Droplet Protein Composition.

    PubMed

    Kory, Nora; Thiam, Abdou-Rachid; Farese, Robert V; Walther, Tobias C

    2015-08-10

    Lipid droplets (LDs) are lipid storage organelles that grow or shrink, depending on the availability of metabolic energy. Proteins recruited to LDs mediate many metabolic functions, including phosphatidylcholine and triglyceride synthesis. How the LD protein composition is tuned to the supply and demand for lipids remains unclear. We show that LDs, in contrast to other organelles, have limited capacity for protein binding. Consequently, macromolecular crowding plays a major role in determining LD protein composition. During lipolysis, when LDs and their surfaces shrink, some, but not all, proteins become displaced. In vitro studies show that macromolecular crowding, rather than changes in monolayer lipid composition, causes proteins to fall off the LD surface. As predicted by a crowding model, proteins compete for binding to the surfaces of LDs. Moreover, the LD binding affinity determines protein localization during lipolysis. Our findings identify protein crowding as an important principle in determining LD protein composition. PMID:26212136

  19. Lipids from the nacreous and prismatic layers of two Pteriomorpha mollusc shells.

    PubMed

    Farre, B; Dauphin, Y

    2009-02-01

    Mollusc shells are acellular biominerals, in which macromolecular structures are intimately associated with mineral phases. Most studies are devoted to proteins, despite sugars have been described. Lipids were extracted from the calcite prismatic and aragonite nacreous layer of two mollusc shells. Fourier Transform Infrared Spectrometry shows that lipids are present in both samples, but they are not similar. Thin layer chromatography confirms that lipids are different in the two studied layers, so that it may be suggested they are species-dependant. Although not yet deciphered, their role in biomineralization and fossilisation processes is probably important.

  20. Biophysical investigations of the structure and function of the tear fluid lipid layer and the effect of ectoine. Part A: natural meibomian lipid films.

    PubMed

    Dwivedi, Mridula; Backers, Hannes; Harishchandra, Rakesh Kumar; Galla, Hans-Joachim

    2014-10-01

    The tear fluid lipid layer is the outermost part of the tear film on the ocular surface which protects the eye from inflammations and injuries. We investigated the influence of ectoine on the structural organization of natural meibomian lipid films using surface activity analysis and topographical studies. These films exhibit a continuous pressure-area isotherm without any phase transition. With the addition of ectoine, the isotherm is expanded towards higher area per molecule values suggesting an increased area occupied by the interfacial lipid molecules. The AFM topology scans of natural meibomian lipid films reveal a presence of fiber-like structures. The addition of ectoine causes an appearance of droplet-like structures which are hypothesized to be tri-acyl-glycerols and other hydrophobic components excluded from the lipid film. Further the material properties of the droplet-like structure with respect to the surrounding were determined by using the quantitative imaging mode of the AFM technique. The droplet-like structures were found to be comparatively softer than the surrounding. Based on the observations a preliminary hypothesis is proposed explaining the mechanism of action of ectoine leading to the fluidization of meibomian lipid films. This suggests the possibility of ectoine as a treatment for the dry eye syndrome.

  1. Early determinants of development: a lipid perspective1234

    PubMed Central

    Carlson, Susan E

    2009-01-01

    This article results from an International Life Sciences Institute workshop on early nutritional determinants of health and development. The presentation on lipids focused mainly on the longer-chain products of the essential fatty acids, particularly docosahexaenoic acid (22:6n–3), and cognitive development as among the most studied lipids and outcomes, respectively, in early human nutrition. Because there have been several recent reviews on this topic, the present review takes a broader perspective with respect to both early development and lipids: an expanded research agenda is plausible on the basis of observations from some human studies and from animal studies. Other lipids known to be provided in variable amounts to infants through human milk are cholesterol and gangliosides. Short sections address the current state of knowledge and some questions that could be pursued. PMID:19321568

  2. Lipid metabolism of commercial layers fed diets containing aflatoxin, fumonisin, and a binder.

    PubMed

    Siloto, E V; Oliveira, E F A; Sartori, J R; Fascina, V B; Martins, B A B; Ledoux, D R; Rottinghaus, G E; Sartori, D R S

    2013-08-01

    Aflatoxins (AF) and fumonisins (FU) are a major problem faced by poultry farmers, leading to huge economic losses. This experiment was conducted to determine the effects of AF (1 mg/kg of feed) and FU (25 mg/kg of feed), singly or in combination, on the lipid metabolism in commercial layers and investigate the efficacy of a commercial binder (2 kg/t of feed) on reducing the toxic effects of these mycotoxins. A total of 168 Hisex Brown layer hens, 37 wk of age, were randomized into a 3 × 2 + 1 factorial arrangement (3 diets with no binder containing AF, FU, and AF+FU; 3 diets with binder containing AF, FU, and AF+FU; and a control diet with no mycotoxins and binders), totaling 7 treatments. The hens contaminated with AF showed the characteristic effects of aflatoxicosis, such as a yellow liver, resulting from the accumulation of liver fat, lower values of plasma very low-density lipoprotein and triglycerides, and higher relative weight of the kidneys and liver. Hepatotoxic and nephrotoxic effects of FU were not observed in this study. On the other hand, the FU caused a reduction in small intestine length and an increase in abdominal fat deposition. The glucan-based binder prevented some of the deleterious effects of these mycotoxins, particularly the effects of AF on hepatic lipid metabolism, kidney relative weight, and FU in the small intestine.

  3. The lipid layer of tears: dependent on meibomian gland function.

    PubMed

    McCulley, James P; Shine, Ward E

    2004-03-01

    There is growing laboratory and clinical evidence implicating the meibomian glands of the eyelid as playing a critical role in the pathogenesis of various ocular surface disorders such as chronic blepharitis and dry eye. Meibomian glands produce a lipid material whose synthesis is dependent on factors such as stem cells, neurological stimulants and hormones. This lipid material is fluid, spreads easily, is a surfactant as well as an aqueous barrier, and must remain functional after a blink. Before delivery it can be modified by factors such as hormone abnormalities and even after delivery it may be modified by lipases produced by ocular bacteria.

  4. Determining the pivotal plane of fluid lipid membranes in simulations

    NASA Astrophysics Data System (ADS)

    Wang, Xin; Deserno, Markus

    2015-10-01

    Each leaflet of a curved lipid membrane contains a surface at which the area strain vanishes, the so-called pivotal plane. Its distance z0 from the bilayer's midplane arises in numerous contexts, for instance the connection between monolayer and bilayer moduli, stress-profile moments, or area-difference elasticity theories. Here, we propose two precise methods for determining the location of the pivotal plane in computer simulations, both of which rely on monitoring the lipid imbalance across a curved bilayer. The first method considers the ratio of lipid number between the two leaflets of cylindrical or spherical vesicles; it hence requires lipid flip-flop for equilibration. The second method looks at the leaflet difference across local sections cut out from a buckled membrane; this observable equilibrates even in the absence of flip-flop. We apply our methods to two different coarse-grained lipid models, the generic three-bead solvent-free Cooke model and a ten-bead representation of dimyristoylphosphocholine with the explicit solvent MARTINI model. The Cooke model is amenable to both methods and gives results that agree at the percent level. Using it, we also show that the pivotal plane moves outward as lipid curvature becomes more positive. The MARTINI model can only be analyzed with the buckling method; the obtained value z0 = 0.850(11) nm lies about 0.4 nm inwards of the glycerol backbone and is hence unexpectedly small. We attribute this to limitations of the coarse-grained description, suggesting that the location of the pivotal plane might be a good indicator for how well lipid models capture the microscopic origins of curvature elasticity. Finally, we also show that the pivotal plane position itself moves as the membrane is bent. The leading correction is linear in curvature, dependent on the Poisson ratio, and can matter when analyzing experimental results obtained from highly curved inverse hexagonal phases.

  5. Lipid bilayer modules as determinants of K+ channel gating.

    PubMed

    Syeda, Ruhma; Santos, Jose S; Montal, Mauricio

    2014-02-14

    The crystal structure of the sensorless pore module of a voltage-gated K(+) (Kv) channel showed that lipids occupy a crevice between subunits. We asked if individual lipid monolayers of the bilayer embody independent modules linked to channel gating modulation. Functional studies using single channel current recordings of the sensorless pore module reconstituted in symmetric and asymmetric lipid bilayers allowed us to establish the deterministic role of lipid headgroup on gating. We discovered that individual monolayers with headgroups that coat the bilayer-aqueous interface with hydroxyls stabilize the channel open conformation. The hydroxyl need not be at a terminal position and the effect is not dependent on the presence of phosphate or net charge on the lipid headgroup. Asymmetric lipid bilayers allowed us to determine that phosphoglycerides with glycerol or inositol on the extracellular facing monolayer stabilize the open conformation of the channel. This indirect effect is attributed to a change in water structure at the membrane interface. By contrast, inclusion of the positively charged lysyl-dioleoyl-phosphatidylglycerol exclusively on the cytoplasmic facing monolayer of the bilayer increases drastically the probability of finding the channel open. Such modulation is mediated by a π-cation interaction between Phe-19 of the pore module and the lysyl moiety anchored to the phosphatidylglycerol headgroup. The new findings imply that the specific chemistry of the lipid headgroup and its selective location in either monolayer of the bilayer dictate the stability of the open conformation of a Kv pore module in the absence of voltage-sensing modules.

  6. Four characteristics and a model of an effective tear film lipid layer (TFLL).

    PubMed

    King-Smith, P Ewen; Bailey, Melissa D; Braun, Richard J

    2013-10-01

    It is proposed that a normal, effective tear film lipid layer (TFLL) should have the following four characteristics: 1) high evaporation resistance to prevent water loss and consequent hyperosmolarity; 2) respreadability, so it will return to its original state after the compression-expansion cycle of the blink; 3) fluidity sufficient to avoid blocking secretion from meibomian glands; 4) gel-like and incompressible structure that can resist forces that may tend to disrupt it. These characteristics tend to be incompatible; for example, lipids that form good evaporation barriers tend to be disrupted by compression-expansion cycles. It is noted that clues about the function and organization of the TFLL can be obtained by comparison with other biological lipid layers, such as lung surfactant and the lipid evaporation barrier of the skin. In an attempt to satisfy the conflicting characteristics, a "multilamellar sandwich model" of the TFLL is proposed, having features in common with the skin evaporation barrier.

  7. Four Characteristics and a Model of an Effective Tear Film Lipid Layer (TFLL)

    PubMed Central

    King-Smith, P. Ewen; Bailey, Melissa D.; Braun, Richard J.

    2015-01-01

    It is proposed that a normal, effective tear film lipid layer (TFLL) should have the following four characteristics: 1) high evaporation resistance to prevent water loss and consequent hyperosmolarity; 2) respreadability, so it will return to its original state after the compression-expansion cycle of the blink; 3) fluidity sufficient to avoid blocking secretion from meibomian glands; 4) gel-like and incompressible structure that can resist forces that may tend to disrupt it. These characteristics tend to be incompatible; for example, lipids that form good evaporation barriers tend to be disrupted by compression-expansion cycles. It is noted that clues about the function and organization of the TFLL can be obtained by comparison with other biological lipid layers, such as lung surfactant and the lipid evaporation barrier of the skin. In an attempt to satisfy the conflicting characteristics, a “multilamellar sandwich model” of the TFLL is proposed, having features in common with the skin evaporation barrier. PMID:24112227

  8. Biomimetic interfaces based on S-layer proteins, lipid membranes and functional biomolecules

    PubMed Central

    Schuster, Bernhard; Sleytr, Uwe B.

    2014-01-01

    Designing and utilization of biomimetic membrane systems generated by bottom-up processes is a rapidly growing scientific and engineering field. Elucidation of the supramolecular construction principle of archaeal cell envelopes composed of S-layer stabilized lipid membranes led to new strategies for generating highly stable functional lipid membranes at meso- and macroscopic scale. In this review, we provide a state-of-the-art survey of how S-layer proteins, lipids and polymers may be used as basic building blocks for the assembly of S-layer-supported lipid membranes. These biomimetic membrane systems are distinguished by a nanopatterned fluidity, enhanced stability and longevity and, thus, provide a dedicated reconstitution matrix for membrane-active peptides and transmembrane proteins. Exciting areas in the (lab-on-a-) biochip technology are combining composite S-layer membrane systems involving specific membrane functions with the silicon world. Thus, it might become possible to create artificial noses or tongues, where many receptor proteins have to be exposed and read out simultaneously. Moreover, S-layer-coated liposomes and emulsomes copying virus envelopes constitute promising nanoformulations for the production of novel targeting, delivery, encapsulation and imaging systems. PMID:24812051

  9. Biophysical investigations of the structure and function of the tear fluid lipid layers and the effect of ectoine. Part B: artificial lipid films.

    PubMed

    Dwivedi, Mridula; Brinkkötter, Marc; Harishchandra, Rakesh Kumar; Galla, Hans-Joachim

    2014-10-01

    The tear fluid lipid layer is present at the outermost part of the tear film which lines the ocular surface and functions to maintain the corneal surface moist by retarding evaporation. Instability in the structure of the tear fluid lipid layer can cause an increased rate of evaporation and thus dry eye syndrome. Ectoine has been previously shown to fluidize lipid monolayers and alter the phase behavior. In the current study we have investigated the effect of ectoine on the artificial tear fluid lipid layer composed of binary and ternary lipid mixtures of dipalmitoyl phosphatidylcholine (DPPC), cholesteryl esters and tri-acyl-glycerols. The focus of our study was mainly the structural and the biophysical aspects of the artificial tear fluid lipid layer using surface activity studies and topology analysis. The presence of ectoine consistently causes an expansion of the pressure-area isotherm indicating increased intermolecular spacing. The topology studies showed the formation of droplet-like structures due to the addition of ectoine only when tri-acyl-glycerol is present in the mixture of DPPC and chol-palmitate, similar to the natural meibomian lipids. Consequently, the hypothesis of an exclusion of tri/di-acyl-glycerol from the meibomian lipid film in the presence of ectoine in the subphase is confirmed. A model describing the effect of ectoine on meibomian lipid films is further presented which may have an application for the use of ectoines in eye drops as a treatment for the dry eye syndrome.

  10. [The lipid layer of the lacrimal tear film: physiology and pathology].

    PubMed

    Lozato, P A; Pisella, P J; Baudouin, C

    2001-06-01

    The preocular tear film (POTF) is composed of a deep aqueous-mucin phase that supports a thin superficial lipid phase. The tear lipid layer (TLL), although thin, stabilizes the POTF providing a 25% surface-tension decrease and a 90-95% aqueous evaporation reduction. TLL is formed from lipids secreted by tarsal meibomian glands and spread onto the ocular surface by blinking. The TLL itself is composed of two phases. A thin and deep polar phase, adjacent to the aqueous-mucin layer, has a surfactant role. A thicker and superficial nonpolar phase has antievaporative properties. At the same time, tear lipocalins help the TLL spread and stabilize the lipid-aqueous interface. For clinical examination, TLL is directly observed with the Tearscope. POTF stability and the evaporation rate depend on the lipid layer pattern. When chronic, POTF qualitative trouble (evaporative syndrome) due to a TLL anomaly, leads to secondary ocular surface impairment with increased tear instability and self-propagation of ocular dryness. Meibomian gland dysfunction (MGD) results from local pathology, dermatologic disease (ocular rosacea) or iatrogenic etiology. Cosmetic use is the other principal cause of TLL destabilization. Lid hygiene is the mainstay of MGD treatment. Systemic antibiotics (cyclins) can be associated in cases of severe symptoms. Topical treatment is useful if there is marginal lid inflammation or infection.

  11. Lipids from the nacreous and prismatic layers of two Pteriomorpha Mollusc shells

    NASA Astrophysics Data System (ADS)

    Farre, B.; Dauphin, Y.

    2009-04-01

    Mollusc shells are the best-known Ca-carbonate biominerals. They are commonly described as a mineralized two layered structure: an outer layer composed of calcite prismatic units, and an internal layer composed of tablets of aragonite: the nacreous layer. An external organic layer (periostracum) is present in most taxa. However, the most common structure in the Mollusc shell is the aragonite crossed lamellar layer, but aragonite prisms, calcite foliated layers and homogeneous layers have been also described by Boggild (1930) in all the Mollusc orders. Since, more detailed descriptions of Bivalve shells have been done (Taylor et al., 1969, 1973). Despite the nacroprismatic arrangement is rare, calcite prismatic and aragonite nacreous layers are the best studied because of their simple 3D structure and large units. Among these Molluscs, some Bivalve species composed of these two layers are of commercial interest, such as the pearl oyster, Pinctada margaritifera, cultivated in French Polynesia to produce black pearls. It is well established that Mollusc shells are composite structures of organic and inorganic components (Hatchett, 1799; Grégoire et al., 1955; Beedham, 1958; Simkiss, 1965; Mutvei, 1969; Cuif et al., 1980; Berman et al., 1993; Kobayashi and Samata, 2006). Numerous studies are concerned with the organic matrix of the shell. Organic components are commonly obtained after a strong or mild decalcification process. They are said to consist of both a soluble and insoluble fraction. The main part of studies is dedicated to the soluble components, and among them, proteins (Grégoire et al., 1955; Grégoire, 1961; Krampitz et al., 1976; Samata et al., 1980, 2004; Weiner, 1983; Miyamoto et al., 2006). Despite the pioneering work of Wada (1980) sugars are usually neglected despite their role in biomineralization. The third component of the organic matrix of calcareous biominerals is lipids. To date, there is a paucity of information concerning the presence

  12. Thermal phase transition behavior of lipid layers on a single human corneocyte cell.

    PubMed

    Imai, Tomohiro; Nakazawa, Hiromitsu; Kato, Satoru

    2013-09-01

    We have improved the selected area electron diffraction method to analyze the dynamic structural change in a single corneocyte cell non-invasively stripped off from human skin surface. The improved method made it possible to obtain reliable diffraction images to trace the structural change in the intercellular lipid layers on a single corneocyte cell during heating from 24°C to 100°C. Comparison of the results with those of synchrotron X-ray diffraction experiments on human stratum corneum sheets revealed that the intercellular lipid layers on a corneocyte cell exhibit essentially the same thermal phase transitions as those in a stratum corneum sheet. These results suggest that the structural features of the lipid layers are well preserved after the mechanical stripping of the corneocyte cell. Moreover, electron diffraction analyses of the thermal phase transition behaviors of the corneocyte cells that had the lipid layers with different distributions of orthorhombic and hexagonal domains at 24°C suggested that small orthorhombic domains interconnected with surrounding hexagonal domains transforms in a continuous manner into new hexagonal domains.

  13. Micro-determinations of alpha-tocopherol in tissue lipids.

    PubMed

    Nishiyama, J; Ellison, E C; Mizuno, G R; Chipault, J R

    1975-01-01

    It has been shown that the determination of small amounts of tocopheral in tissue lipids by methods that use, in sequence, saponification, extraction, TLC and colorimetric assay, is greatly influenced by the concentration of soap in the saponification mixture. Soaps appear to interfere with the recovery of tocopherols during the subsequent extraction step. Based on this knowledge a method is described that was used to measure as little as 15 to 20 ng of tocopherol in 3 to 4 mg of lipid obtained from rat retinas. Presumably the method would be applicable to other tissues as well. The results show that the rat retina is readily depleted of tocopherol if none is included in the diet.

  14. Lipids from the nacreous and prismatic layers of two Pteriomorpha Mollusc shells

    NASA Astrophysics Data System (ADS)

    Farre, B.; Dauphin, Y.

    2009-04-01

    Mollusc shells are the best-known Ca-carbonate biominerals. They are commonly described as a mineralized two layered structure: an outer layer composed of calcite prismatic units, and an internal layer composed of tablets of aragonite: the nacreous layer. An external organic layer (periostracum) is present in most taxa. However, the most common structure in the Mollusc shell is the aragonite crossed lamellar layer, but aragonite prisms, calcite foliated layers and homogeneous layers have been also described by Boggild (1930) in all the Mollusc orders. Since, more detailed descriptions of Bivalve shells have been done (Taylor et al., 1969, 1973). Despite the nacroprismatic arrangement is rare, calcite prismatic and aragonite nacreous layers are the best studied because of their simple 3D structure and large units. Among these Molluscs, some Bivalve species composed of these two layers are of commercial interest, such as the pearl oyster, Pinctada margaritifera, cultivated in French Polynesia to produce black pearls. It is well established that Mollusc shells are composite structures of organic and inorganic components (Hatchett, 1799; Grégoire et al., 1955; Beedham, 1958; Simkiss, 1965; Mutvei, 1969; Cuif et al., 1980; Berman et al., 1993; Kobayashi and Samata, 2006). Numerous studies are concerned with the organic matrix of the shell. Organic components are commonly obtained after a strong or mild decalcification process. They are said to consist of both a soluble and insoluble fraction. The main part of studies is dedicated to the soluble components, and among them, proteins (Grégoire et al., 1955; Grégoire, 1961; Krampitz et al., 1976; Samata et al., 1980, 2004; Weiner, 1983; Miyamoto et al., 2006). Despite the pioneering work of Wada (1980) sugars are usually neglected despite their role in biomineralization. The third component of the organic matrix of calcareous biominerals is lipids. To date, there is a paucity of information concerning the presence

  15. Lipid modification gives rise to two distinct Haloferax volcanii S-layer glycoprotein populations.

    PubMed

    Kandiba, Lina; Guan, Ziqiang; Eichler, Jerry

    2013-03-01

    The S-layer glycoprotein is the sole component of the protein shell surrounding Haloferax volcanii cells. The deduced amino acid sequence of the S-layer glycoprotein predicts the presence of a C-terminal membrane-spanning domain. However, several earlier observations, including the ability of EDTA to selectively solubilize the protein, are inconsistent with the presence of a trans-membrane sequence. In the present report, sequential solubilization of the S-layer glycoprotein by EDTA and then with detergent revealed the existence of two distinct populations of the S-layer glycoprotein. Whereas both S-layer glycoprotein populations underwent signal peptide cleavage and N-glycosylation, base hydrolysis followed by mass spectrometry revealed that a lipid, likely archaetidic acid, modified only the EDTA-solubilized version of the protein. These observations are consistent with the S-layer glycoprotein being initially synthesized as an integral membrane protein and subsequently undergoing a processing event in which the extracellular portion of the protein is separated from the membrane-spanning domain and transferred to a waiting lipid moiety.

  16. Characterization of Floating Surface Layers of Lipids and Lipopolymers by Surface-Sensitive Scattering

    NASA Astrophysics Data System (ADS)

    Krüger, Peter; Lösche, Mathias

    Nanotechnology and molecular (bio-)engineering are making ever deepening inroads into everybodys daily life. Physicochemical and biotechnological achievements in the design of physiologically active supramolecular assemblies have brought about the quest for their submolecular-level characterization. We employ surface-sensitive scattering techniques for the investigation of planar lipid membranes - floating monolayers on aqueous surfaces - to correlate structural, functional and dynamic aspects of biomembrane models. This chapter surveys recent work on the submolecular structure of floating phospholipid monolayers - where the advent of third-generation synchrotron X-ray sources has driven the development of realistic, submolecular-scale quasi-chemical models - as well as of more complex systems: cation binding to anionic lipid surfaces; conformational changes of lipopolymers undergoing phase transitions; the conformational organization of phosphatidylinositol and phosphatidylinositides, as examples of physiologically important lipids; and the adsorption of peptides (neuropeptide Y, NPY) or solvents (dimethylsulfoxide, DMSO) onto phospholipid surface layers.

  17. Probing Peptide and Protein Insertion in a Biomimetic S-Layer Supported Lipid Membrane Platform

    PubMed Central

    Damiati, Samar; Schrems, Angelika; Sinner, Eva-Kathrin; Sleytr, Uwe B.; Schuster, Bernhard

    2015-01-01

    The most important aspect of synthetic lipid membrane architectures is their ability to study functional membrane-active peptides and membrane proteins in an environment close to nature. Here, we report on the generation and performance of a biomimetic platform, the S-layer supported lipid membrane (SsLM), to investigate the structural and electrical characteristics of the membrane-active peptide gramicidin and the transmembrane protein α-hemolysin in real-time using a quartz crystal microbalance with dissipation monitoring in combination with electrochemical impedance spectroscopy. A shift in membrane resistance is caused by the interaction of α-hemolysin and gramicidin with SsLMs, even if only an attachment onto, or functional channels through the lipid membrane, respectively, are formed. Moreover, the obtained results did not indicate the formation of functional α-hemolysin pores, but evidence for functional incorporation of gramicidin into this biomimetic architecture is provided. PMID:25633104

  18. Development of "all natural" layer-by-layer redispersible solid lipid nanoparticles by nano spray drying technology.

    PubMed

    Wang, Taoran; Hu, Qiaobin; Zhou, Mingyong; Xia, Yan; Nieh, Mu-Ping; Luo, Yangchao

    2016-10-01

    Solid lipid nanoparticles (SLNs) have gained tremendous attraction as carriers for controlled drug delivery. Despite numerous advances in the field, one long-standing historical challenge for their practical applications remains unmet: redispersibility after drying. In this work, a novel design of SLNs using a layer-by-layer (LbL) technique was developed and the formulations were optimized by surface response methodology (Box-Behnken design). To the best of our knowledge, this is the first study reporting the fabrication of SLNs from all natural ingredients in the absence of any synthetic surfactants or coatings. The SLNs were prepared by a combined solvent-diffusion and hot homogenization method, with soy lecithin as natural emulsifier (first layer), followed by the subsequent coating with sodium caseinate (second layer) and pectin (third layer), both of which are natural food biopolymers. The adsorption of pectin coating onto caseinate was reinforced by hydrophobic and electrostatic interactions induced by a pH-driven process along with thermal treatment. The innovative nano spray drying technology was further explored to obtain ultra-fine powders of SLNs. Compared to uncoated or single-layer coated SLNs powders, which showed severe aggregation after spray drying, the well-separated particles with spherical shape and smooth surface were obtained for layer-by-layer (LbL) SLNs, which were redispersible into water without variation of dimension, shape and morphology. The SLNs were characterized by Fourier transform infrared and high-performance differential scanning calorimetry for their physical properties. The LbL-coated SLNs based on all natural ingredients have promising features for future applications as drug delivery systems, overcoming the major obstacles in conventional spray drying and redispersing SLNs-based formulations. PMID:27470922

  19. Development of "all natural" layer-by-layer redispersible solid lipid nanoparticles by nano spray drying technology.

    PubMed

    Wang, Taoran; Hu, Qiaobin; Zhou, Mingyong; Xia, Yan; Nieh, Mu-Ping; Luo, Yangchao

    2016-10-01

    Solid lipid nanoparticles (SLNs) have gained tremendous attraction as carriers for controlled drug delivery. Despite numerous advances in the field, one long-standing historical challenge for their practical applications remains unmet: redispersibility after drying. In this work, a novel design of SLNs using a layer-by-layer (LbL) technique was developed and the formulations were optimized by surface response methodology (Box-Behnken design). To the best of our knowledge, this is the first study reporting the fabrication of SLNs from all natural ingredients in the absence of any synthetic surfactants or coatings. The SLNs were prepared by a combined solvent-diffusion and hot homogenization method, with soy lecithin as natural emulsifier (first layer), followed by the subsequent coating with sodium caseinate (second layer) and pectin (third layer), both of which are natural food biopolymers. The adsorption of pectin coating onto caseinate was reinforced by hydrophobic and electrostatic interactions induced by a pH-driven process along with thermal treatment. The innovative nano spray drying technology was further explored to obtain ultra-fine powders of SLNs. Compared to uncoated or single-layer coated SLNs powders, which showed severe aggregation after spray drying, the well-separated particles with spherical shape and smooth surface were obtained for layer-by-layer (LbL) SLNs, which were redispersible into water without variation of dimension, shape and morphology. The SLNs were characterized by Fourier transform infrared and high-performance differential scanning calorimetry for their physical properties. The LbL-coated SLNs based on all natural ingredients have promising features for future applications as drug delivery systems, overcoming the major obstacles in conventional spray drying and redispersing SLNs-based formulations.

  20. Thermal stability of ladderane lipids as determined by hydrous pyrolysis

    USGS Publications Warehouse

    Jaeschke, A.; Lewan, M.D.; Hopmans, E.C.; Schouten, S.; Sinninghe, Damste J.S.

    2008-01-01

    Anaerobic ammonium oxidation (anammox) has been recognized as a major process resulting in loss of fixed inorganic nitrogen in the marine environment. Ladderane lipids, membrane lipids unique to anammox bacteria, have been used as markers for the detection of anammox in marine settings. However, the fate of ladderane lipids after sediment burial and maturation is unknown. In this study, anammox bacterial cell material was artificially matured by hydrous pyrolysis at constant temperatures ranging from 120 to 365 ??C for 72 h to study the stability of ladderane lipids during progressive dia- and catagenesis. HPLC-MS/MS analysis revealed that structural alterations of ladderane lipids already occurred at 120 ??C. At temperatures >140 ??C, ladderane lipids were absent and only more thermally stable products could be detected, i.e., ladderane derivatives in which some of the cyclobutane rings were opened. These diagenetic products of ladderane lipids were still detectable up to temperatures of 260 ??C using GC-MS. Thus, ladderane lipids are unlikely to occur in ancient sediments and sedimentary rocks, but specific diagenetic products of ladderane lipids will likely be present in sediments and sedimentary rocks of relatively low maturity (i.e., C31 hopane 22S/(22S + 22R) ratio 0.5). ?? 2008 Elsevier Ltd.

  1. Spectroscopic study of 3-Hydroxyflavone - protein interaction in lipidic bi-layers immobilized on silver nanoparticles.

    PubMed

    Voicescu, Mariana; Ionescu, Sorana; Nistor, Cristina L

    2017-01-01

    The interaction of 3-Hydroxyflavone with serum proteins (BSA and HSA) in lecithin lipidic bi-layers (PC) immobilized on silver nanoparticles (SNPs), was studied by fluorescence and Raman spectroscopy. BSA secondary structure was quantified with a deconvolution algorithm, showing a decrease in α-helix structure when lipids were added to the solution. The effect of temperature on the rate of the excited-state intra-molecular proton transfer and on the dual fluorescence emission of 3-HF in the HSA/PC/SNPs systems was discussed. Evaluation of the antioxidant activity of 3-HF in HSA/PC/SNPs systems was also studied. The antioxidant activity of 3-HF decreased in the presence of SNPs. The results are discussed with relevance to the secondary structure of proteins and of the 3-HF based nano-systems to a topical formulation useful in the oxidative stress process. PMID:27380623

  2. Intraspecific Differences in Lipid Content of Calanoid Copepods across Fine-Scale Depth Ranges within the Photic Layer

    PubMed Central

    Zarubin, Margarita; Farstey, Viviana; Wold, Anette; Falk-Petersen, Stig; Genin, Amatzia

    2014-01-01

    Copepods are among the most abundant and diverse groups of mesozooplankton in the world's oceans. Each species has a certain depth range within which different individuals (of the same life stage and sex) are found. Lipids are accumulated in many calanoid copepods for energy storage and reproduction. Lipid content in some species increases with depth, however studies so far focused mostly on temperate and high-latitude seasonal vertically migrating copepods and compared lipid contents among individuals either from coarse layers or between diapausing, deep-dwelling copepods and individuals found in the photic, near-surface layer. Here we examined whether lipid contents of individual calanoid copepods of the same species, life stage/sex differ between finer depth layers within the upper water column of subtropical and Arctic seas. A total of 6 calanoid species were collected from samples taken at precise depths within the photic layer in both cold eutrophic and warm oligotrophic environments using SCUBA diving, MOCNESS and Multinet. Measurements of lipid content were obtained from digitized photographs of the collected individuals. The results revealed significant differences in lipid content across depth differences as small as 12–15 meters for Mecynocera clausi C5 and Ctenocalanus vanus C5 (Red Sea), Clausocalanus furcatus males and two clausocalanid C5s (Mediterranean Sea), and Calanus glacialis C5 (Arctic). We suggest two possible explanations for the differences in lipid content with depth on such a fine scale: predator avoidance and buoyancy. PMID:24667529

  3. Intraspecific differences in lipid content of calanoid copepods across fine-scale depth ranges within the photic layer.

    PubMed

    Zarubin, Margarita; Farstey, Viviana; Wold, Anette; Falk-Petersen, Stig; Genin, Amatzia

    2014-01-01

    Copepods are among the most abundant and diverse groups of mesozooplankton in the world's oceans. Each species has a certain depth range within which different individuals (of the same life stage and sex) are found. Lipids are accumulated in many calanoid copepods for energy storage and reproduction. Lipid content in some species increases with depth, however studies so far focused mostly on temperate and high-latitude seasonal vertically migrating copepods and compared lipid contents among individuals either from coarse layers or between diapausing, deep-dwelling copepods and individuals found in the photic, near-surface layer. Here we examined whether lipid contents of individual calanoid copepods of the same species, life stage/sex differ between finer depth layers within the upper water column of subtropical and Arctic seas. A total of 6 calanoid species were collected from samples taken at precise depths within the photic layer in both cold eutrophic and warm oligotrophic environments using SCUBA diving, MOCNESS and Multinet. Measurements of lipid content were obtained from digitized photographs of the collected individuals. The results revealed significant differences in lipid content across depth differences as small as 12-15 meters for Mecynocera clausi C5 and Ctenocalanus vanus C5 (Red Sea), Clausocalanus furcatus males and two clausocalanid C5s (Mediterranean Sea), and Calanus glacialis C5 (Arctic). We suggest two possible explanations for the differences in lipid content with depth on such a fine scale: predator avoidance and buoyancy. PMID:24667529

  4. Intraspecific differences in lipid content of calanoid copepods across fine-scale depth ranges within the photic layer.

    PubMed

    Zarubin, Margarita; Farstey, Viviana; Wold, Anette; Falk-Petersen, Stig; Genin, Amatzia

    2014-01-01

    Copepods are among the most abundant and diverse groups of mesozooplankton in the world's oceans. Each species has a certain depth range within which different individuals (of the same life stage and sex) are found. Lipids are accumulated in many calanoid copepods for energy storage and reproduction. Lipid content in some species increases with depth, however studies so far focused mostly on temperate and high-latitude seasonal vertically migrating copepods and compared lipid contents among individuals either from coarse layers or between diapausing, deep-dwelling copepods and individuals found in the photic, near-surface layer. Here we examined whether lipid contents of individual calanoid copepods of the same species, life stage/sex differ between finer depth layers within the upper water column of subtropical and Arctic seas. A total of 6 calanoid species were collected from samples taken at precise depths within the photic layer in both cold eutrophic and warm oligotrophic environments using SCUBA diving, MOCNESS and Multinet. Measurements of lipid content were obtained from digitized photographs of the collected individuals. The results revealed significant differences in lipid content across depth differences as small as 12-15 meters for Mecynocera clausi C5 and Ctenocalanus vanus C5 (Red Sea), Clausocalanus furcatus males and two clausocalanid C5s (Mediterranean Sea), and Calanus glacialis C5 (Arctic). We suggest two possible explanations for the differences in lipid content with depth on such a fine scale: predator avoidance and buoyancy.

  5. Phospatidylserine or ganglioside--which of anionic lipids determines the effect of cationic dextran on lipid membrane?

    PubMed

    Hąc-Wydro, Katarzyna; Wydro, Paweł; Cetnar, Andrzej; Włodarczyk, Grzegorz

    2015-02-01

    In this work the influence of cationic polymer, namely diethylaminoethyl DEAE-dextran on model lipid membranes was investigated. This polymer is of a wide application as a biomaterial and a drug carrier and its cytotoxicity toward various cancer cells was also confirmed. It was suggested that anticancer effect of cationic dextran is connected with the binding of the polymer to the negatively charged sialic acid residues overexpressed in cancer membrane. This fact encouraged us to perform the studies aimed at verifying whether the effect of cationic DEAE-dextran on membrane is determined only by the presence of the negatively charged lipid in the system or the kind of anionic lipid is also important. To reach this goal systematic investigations on the effect of dextran on various one-component lipid monolayers and multicomponent hepatoma cell model membranes differing in the level and the kind of anionic lipids (phosphatidylserine, sialic acid-containing ganglioside GM3 or their mixture) were done. As evidenced the results the effect of DEAE-dextran on the model system is determined by anionic lipid-polymer electrostatic interactions. However, the magnitude of the effect of cationic polymer is strongly dependent on the kind of anionic lipid in the model system. Namely, the packing and ordering of the mixtures containing ganglioside GM3 were more affected by DEAE-dextran than phosphatidylserine-containing monolayers. Although the experiments were done on model systems and therefore further studies are highly needed, the collected data may indicate that ganglioside may be important in the differentiation of the effect of cationic dextran on membranes. PMID:25576813

  6. Line active hybrid lipids determine domain size in phase separation of saturated and unsaturated lipids.

    PubMed

    Brewster, Robert; Safran, Samuel A

    2010-03-17

    A simple model of the line activity of a hybrid lipid (e.g., POPC) with one fully saturated chain and one partially unsaturated chain demonstrates that these lipids preferentially pack at curved interfaces between phase-separated saturated and unsaturated domains. We predict that the domain sizes typically range from tens to hundreds of nm, depending on molecular interactions and parameters such as molecular volume and area per headgroup in the bulk fluid phase. The role of cholesterol is taken into account by an effective change in the headgroup areas and the domain sizes are predicted to increase with cholesterol concentration.

  7. Line Active Hybrid Lipids Determine Domain Size in Phase Separation of Saturated and Unsaturated Lipids

    PubMed Central

    Brewster, Robert; Safran, Samuel A.

    2010-01-01

    A simple model of the line activity of a hybrid lipid (e.g., POPC) with one fully saturated chain and one partially unsaturated chain demonstrates that these lipids preferentially pack at curved interfaces between phase-separated saturated and unsaturated domains. We predict that the domain sizes typically range from tens to hundreds of nm, depending on molecular interactions and parameters such as molecular volume and area per headgroup in the bulk fluid phase. The role of cholesterol is taken into account by an effective change in the headgroup areas and the domain sizes are predicted to increase with cholesterol concentration. PMID:20303848

  8. The lymph lipid precursor pool is a key determinant of intestinal lymphatic drug transport.

    PubMed

    Trevaskis, Natalie L; Porter, Christopher J H; Charman, William N

    2006-02-01

    The influence of the size and turnover kinetics of the enterocyte-based lymph lipid precursor pool (LLPP) on intestinal lymphatic drug transport has been examined. Mesenteric lymph duct-cannulated rats were infused intraduodenally with low (2-5 mg/h) or high (20 mg/h) lipid-dose formulations containing 100 microg/h halofantrine (Hf, a model drug) and 1 microCi/h (14)C-oleic acid (OA) (as a marker for lipid transport) until steady-state rates of lipid(dX(L)/dt)(ss) and drug (dD(L)/dt)(ss) transport in lymph were obtained. After 5 h, the infusion was changed to formulations of the same composition but excluding (14)C-OA and Hf, allowing calculation of the first order rate constants describing turnover of lipid (K(X)) and drug (K(D)) from the LLPP into the lymph from the washout kinetics. The mass of lipid (X(LP)) and drug (D(LP)) in the LLPP was also determined. Biliary-lipid output was determined in a separate group of rats that had been infused with the same formulations. The results indicate that after administration of high lipid doses, lymphatic drug transport is dependent on the mass of exogenous lipid available in the LLPP and the rate of lipid pool turnover into the lymph. In contrast, after administration of low lipid doses, biliary-derived endogenous lipids are most likely to be the primary drivers of drug incorporation into the LLPP and lymph. Therefore, the LLPP size and composition seem to be major determinants of lymphatic drug transport, and formulation components, which increase lipid pool size, may therefore enhance lymphatic drug transport. PMID:16249368

  9. Glass transition temperature of water confined in lipid membranes as determined by anelastic spectroscopy

    NASA Astrophysics Data System (ADS)

    Castellano, C.; Generosi, J.; Congiu, A.; Cantelli, R.

    2006-12-01

    The research of gene delivery vehicles used in gene therapy is focused on nonviral vectors like lipid membranes. Such vectors, nonimmunogenic and biodegradable, are formed by complexation of DNA with a mixture of cationic lipids and a neutral colipid which improve the transfection efficiency. A main topic related to lipid membrane dynamics is their capability to spontaneously confine water. At present the value of the glass transition temperature (Tg) is largely debated and determined only by some indirect methods. Here the authors show that anelastic spectroscopy allows the confined water Tg value to be directly identified in several lipid mixtures.

  10. Purification and lipid-layer crystallization of yeast RNA polymerase II.

    PubMed Central

    Edwards, A M; Darst, S A; Feaver, W J; Thompson, N E; Burgess, R R; Kornberg, R D

    1990-01-01

    Yeast RNA polymerase II was purified to homogeneity by a rapid procedure involving immunoaffinity chromatography. The purified enzyme contained 10 subunits, as reported for conventional preparations, but with no detectable proteolysis of the largest subunit. In assays of initiation of transcription at the yeast CYC1 promoter, the enzyme complemented the deficiency of an extract from a strain that produces a temperature-sensitive polymerase II. Mammalian RNA polymerase II was inactive in this initiation assay. The purified yeast enzyme formed two-dimensional crystals on positively charged lipid layers, as previously found for Escherichia coli RNA polymerase holoenzyme. Image analysis of electron micrographs of crystals in negative stain, which diffracted to about 30-A resolution, showed protein densities of dimensions consistent with those of single polymerase molecules. Images PMID:2179949

  11. Engineering and validation of a novel lipid thin film for biomembrane modeling in lipophilicity determination of drugs and xenobiotics

    PubMed Central

    Idowu, Sunday Olakunle; Adeyemo, Morenikeji Ambali; Ogbonna, Udochi Ihechiluru

    2009-01-01

    Background Determination of lipophilicity as a tool for predicting pharmacokinetic molecular behavior is limited by the predictive power of available experimental models of the biomembrane. There is current interest, therefore, in models that accurately simulate the biomembrane structure and function. A novel bio-device; a lipid thin film, was engineered as an alternative approach to the previous use of hydrocarbon thin films in biomembrane modeling. Results Retention behavior of four structurally diverse model compounds; 4-amino-3,5-dinitrobenzoic acid (ADBA), naproxen (NPX), nabumetone (NBT) and halofantrine (HF), representing 4 broad classes of varying molecular polarities and aqueous solubility behavior, was investigated on the lipid film, liquid paraffin, and octadecylsilane layers. Computational, thermodynamic and image analysis confirms the peculiar amphiphilic configuration of the lipid film. Effect of solute-type, layer-type and variables interactions on retention behavior was delineated by 2-way analysis of variance (ANOVA) and quantitative structure property relationships (QSPR). Validation of the lipid film was implemented by statistical correlation of a unique chromatographic metric with Log P (octanol/water) and several calculated molecular descriptors of bulk and solubility properties. Conclusion The lipid film signifies a biomimetic artificial biological interface capable of both hydrophobic and specific electrostatic interactions. It captures the hydrophilic-lipophilic balance (HLB) in the determination of lipophilicity of molecules unlike the pure hydrocarbon film of the prior art. The potentials and performance of the bio-device gives the promise of its utility as a predictive analytic tool for early-stage drug discovery science. PMID:19735551

  12. The real reason for having a meibomian lipid layer covering the outer surface of the tear film - A review.

    PubMed

    Millar, Thomas J; Schuett, Burkhardt S

    2015-08-01

    This review critically evaluates a broad range of literature in order to show the relationship between meibum, tear lipids and the tear film lipid layer (TFLL). The relationship of meibum composition to dry eye syndrome is briefly discussed. The review also explores the interactions between aqueous and the TFLL by examining the correlations between meibomian lipids and lipids extracted from whole tears, and by considering protein adsorption to the TFLL from the aqueous. Although it is clear to the authors that a normal tear film resists evaporation, an emerging idea from the literature is that the main purpose of the TFLL is to allow the spread of the tear film and to prevent its collapse onto the ocular surface, rather than to be an evaporative blanket. Current models on the possible structure of the TFLL are also examined.

  13. The real reason for having a meibomian lipid layer covering the outer surface of the tear film - A review.

    PubMed

    Millar, Thomas J; Schuett, Burkhardt S

    2015-08-01

    This review critically evaluates a broad range of literature in order to show the relationship between meibum, tear lipids and the tear film lipid layer (TFLL). The relationship of meibum composition to dry eye syndrome is briefly discussed. The review also explores the interactions between aqueous and the TFLL by examining the correlations between meibomian lipids and lipids extracted from whole tears, and by considering protein adsorption to the TFLL from the aqueous. Although it is clear to the authors that a normal tear film resists evaporation, an emerging idea from the literature is that the main purpose of the TFLL is to allow the spread of the tear film and to prevent its collapse onto the ocular surface, rather than to be an evaporative blanket. Current models on the possible structure of the TFLL are also examined. PMID:25981748

  14. Genetic determinants of obesity-related lipid traits.

    PubMed

    Sonnenberg, Gabriele E; Krakower, Glenn R; Martin, Lisa J; Olivier, Michael; Kwitek, Anne E; Comuzzie, Anthony G; Blangero, John; Kissebah, Ahmed H

    2004-04-01

    In our ongoing effort to identify genes influencing the biological pathways that underlie the metabolic disturbances associated with obesity, we performed genome-wide scanning in 2,209 individuals distributed over 507 Caucasian families to localize quantitative trait loci (QTLs), which affect variation of plasma lipids. Pedigree-based analysis using a quantitative trait variance component linkage method that localized a QTL on chromosome 7q35-q36, which linked to variation in levels of plasma triglyceride [TG, logarithm of odds (LOD) score = 3.7] and was suggestive of linkage to LDL-cholesterol (LDL-C, LOD = 2.2). Covariates of the TG linkage included waist circumference, fasting insulin, and insulin:glucose, but not body mass index or hip circumference. Plasma HDL-cholesterol (HDL-C) levels were suggestively linked to a second QTL on chromosome 12p12.3 (LOD = 2.6). Five other QTLs with lower LOD scores were identified for plasma levels of LDL-C, HDL-C, and total cholesterol. These newly identified loci likely harbor genetic elements that influence traits underlying lipid adversities associated with obesity. PMID:14754912

  15. Determination of partition coefficient of spin probe between different lipid membrane phases.

    PubMed

    Arsov, Zoran; Strancar, Janez

    2005-01-01

    Model lipid membranes made from binary mixtures of dimyristoylphosphatidylcholine/dipalmitoylphosphatidylcholine (DMPC/DPPC) and dimyristoylphosphatidylcholine/cholesterol (DMPC/Chol) exhibit coexistence of diverse lipid phases at appropriate temperature and composition. Since lipids in different phases show different structural and motional properties, it is expected that the corresponding spin probe electron paramagnetic resonance (EPR) spectra will be superposition of several spectral components. From comparison of proportions of spectral components of the EPR spectrum with the fractions of the corresponding lipid phases obtained from known phase diagrams the partition coefficient of spin probe methyl ester of 5-doxyl palmitate between different lipid phases was determined. The results indicate that the used spin probe partitions approximately equally between different phases. PMID:16309270

  16. Fabrication of fracture-free nanoglassified substrates by layer-by-layer deposition with a paint gun technique for real-time monitoring of protein-lipid interactions.

    PubMed

    Linman, Matthew J; Culver, Sean P; Cheng, Quan

    2009-03-01

    New sensing materials that are robust, biocompatible, and amenable to array fabrication are vital to the development of novel bioassays. Herein we report the fabrication of ultrathin (ca. 5-8 nm) glass (silicate) layers on top of a gold surface for surface plasmon resonance (SPR) biosensing applications. The nanoglass layers are fabricated by layer-by-layer (LbL) deposition of poly(allylamine) hydrochloride (PAH) and sodium silicate (SiO(x)), followed by calcination at high temperature. To deposit these layers in a uniform and reproducible manner, we employed a high-volume, low-pressure (HVLP) paint gun technique that offers high precision and better control through pressurized nitrogen gas. The new substrates are stable in solution for a long period of time, and scanning electron microscopy (SEM) images confirm that these films are nearly fracture-free. In addition, atomic force microscopy (AFM) indicates that the surface roughness of the silicate layers is low (rms = 2 to 3 nm), similar to that of bare glass slides. By tuning the experimental parameters such as HVLP gun pressure and layers deposited, different surface morphology could be obtained as revealed by fluorescence microscopy and SEM images. To demonstrate the utility of these ultrathin, fracture-free substrates, lipid bilayer membranes composed of phosphorylated derivatives of phosphoinositides (PIs) were deposited on the new substrates for biosensing applications. Fluorescence recovery after photobleaching (FRAP) data indicated that these lipid components in the membranes were highly mobile. Furthermore, interactions of PtdIns(4,5)P2 and PtdIns(4)P lipids with their respective binding proteins were detected with high sensitivity by using SPR spectroscopy. This method of glass deposition can be combined with already well-developed surface chemistry for a range of planar glass assay applications, and the process is amenable to automation for mass production of nanometer thick silicate chips in a highly

  17. Statistical comparison of classifiers applied to the interferential tear film lipid layer automatic classification.

    PubMed

    Remeseiro, B; Penas, M; Mosquera, A; Novo, J; Penedo, M G; Yebra-Pimentel, E

    2012-01-01

    The tear film lipid layer is heterogeneous among the population. Its classification depends on its thickness and can be done using the interference pattern categories proposed by Guillon. The interference phenomena can be characterised as a colour texture pattern, which can be automatically classified into one of these categories. From a photography of the eye, a region of interest is detected and its low-level features are extracted, generating a feature vector that describes it, to be finally classified in one of the target categories. This paper presents an exhaustive study about the problem at hand using different texture analysis methods in three colour spaces and different machine learning algorithms. All these methods and classifiers have been tested on a dataset composed of 105 images from healthy subjects and the results have been statistically analysed. As a result, the manual process done by experts can be automated with the benefits of being faster and unaffected by subjective factors, with maximum accuracy over 95%.

  18. Biomimetic biosensor based on lipidic layers containing tyrosinase and lutetium bisphthalocyanine for the detection of antioxidants.

    PubMed

    Apetrei, C; Alessio, P; Constantino, C J L; de Saja, J A; Rodriguez-Mendez, M L; Pavinatto, F J; Ramos Fernandes, E Giuliani; Zucolotto, V; Oliveira, O N

    2011-01-15

    This paper describes the preparation of a biomimetic Langmuir-Blodgett film of tyrosinase incorporated in a lipidic layer and the use of lutetium bisphthalocyanine as an electron mediator for the voltammetric detection of phenol derivatives, which include one monophenol (vanillic acid), two diphenols (catechol and caffeic acid) and two triphenols (gallic acid and pyrogallol). The first redox process of the voltammetric responses is associated with the reduction of the enzymatically formed o-quinone and is favoured by the lutetium bisphthalocyanine because significant signal amplification is observed, while the second is associated with the electrochemical oxidation of the antioxidant and occurs at lower potentials in the presence of an electron mediator. The biosensor shows low detection limit (1.98×10(-6)-27.49×10(-6) M), good reproducibility, and high affinity to antioxidants (K(M) in the range of 62.31-144.87 μM). The excellent functionality of the enzyme obtained using a biomimetic immobilisation method, the selectivity afforded by enzyme catalysis, the signal enhancement caused by the lutetium bisphthalocyanine mediator and the increased selectivity of the curves due to the occurrence of two redox processes make these sensors exceptionally suitable for the detection of phenolic compounds.

  19. Lipids that determine detergent resistance of MDCK cell membrane fractions.

    PubMed

    Manni, Marco M; Cano, Ainara; Alonso, Cristina; Goñi, Félix M

    2015-10-01

    A comparative lipidomic study has been performed of whole Madin-Darby canine kidney epithelial cells and of the detergent-resistant membrane fraction (DRM) obtained after treating the cells with the non-ionic detergent Triton X-100. The DRM were isolated following a standard procedure that is extensively used in cell biology studies. Significant differences were found in the lipid composition of the whole cells and of DRM. The latter were enriched in all the analyzed sphingolipid classes: sphingomyelins, ceramides and hexosylceramides. Diacylglycerols were also preferentially found in DRM. The detergent-resistant fraction was also enriched in saturated over unsaturated fatty acyl chains, and in sn-1 acyl chains containing 16 carbon atoms, over the longer and shorter ones. The glycerophospholipid species phosphatidylethanolamines and phosphatidylinositols, that were mainly unsaturated, did not show a preference for DRM. Phosphatidylcholines were an intermediate case: the saturated, but not the unsaturated species were found preferentially in DRM. The question remains on whether these DRM, recovered from detergent-membrane mixtures by floatation over a sucrose gradient, really correspond to membrane domains existing in the cell membrane prior to detergent treatment.

  20. Lipids that determine detergent resistance of MDCK cell membrane fractions.

    PubMed

    Manni, Marco M; Cano, Ainara; Alonso, Cristina; Goñi, Félix M

    2015-10-01

    A comparative lipidomic study has been performed of whole Madin-Darby canine kidney epithelial cells and of the detergent-resistant membrane fraction (DRM) obtained after treating the cells with the non-ionic detergent Triton X-100. The DRM were isolated following a standard procedure that is extensively used in cell biology studies. Significant differences were found in the lipid composition of the whole cells and of DRM. The latter were enriched in all the analyzed sphingolipid classes: sphingomyelins, ceramides and hexosylceramides. Diacylglycerols were also preferentially found in DRM. The detergent-resistant fraction was also enriched in saturated over unsaturated fatty acyl chains, and in sn-1 acyl chains containing 16 carbon atoms, over the longer and shorter ones. The glycerophospholipid species phosphatidylethanolamines and phosphatidylinositols, that were mainly unsaturated, did not show a preference for DRM. Phosphatidylcholines were an intermediate case: the saturated, but not the unsaturated species were found preferentially in DRM. The question remains on whether these DRM, recovered from detergent-membrane mixtures by floatation over a sucrose gradient, really correspond to membrane domains existing in the cell membrane prior to detergent treatment. PMID:26320877

  1. Selective analysis of lipids by thin-layer chromatography blot matrix-assisted laser desorption/ionization imaging mass spectrometry.

    PubMed

    Zaima, Nobuhiro; Goto-Inoue, Naoko; Adachi, Kohsuke; Setou, Mitsutoshi

    2011-01-01

    Thin-layer chromatography (TLC) is an essential method for food composition analyses such as lipid nutrition analysis. TLC can be used to obtain information about the lipid composition of foods; however, it cannot be used for analyses at the molecular level. Recently we developed a new method that combines matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) with TLC-blotting (TLC-Blot-MALDI-IMS). The combination of MALDI-IMS and TLC blotting enabled detailed and sensitive analyses of lipids. In this study, we applied TLC-Blot-MALDI-IMS for analysis of major phospholipids extracted from bluefin tuna. We showed that TLC-Blot-MALDI-IMS analysis could visualize and identify major phospholipids such as phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin.

  2. Structural determinants of protein partitioning into ordered membrane domains and lipid rafts.

    PubMed

    Lorent, Joseph Helmuth; Levental, Ilya

    2015-11-01

    Increasing evidence supports the existence of lateral nanoscopic lipid domains in plasma membranes, known as lipid rafts. These domains preferentially recruit membrane proteins and lipids to facilitate their interactions and thereby regulate transmembrane signaling and cellular homeostasis. The functionality of raft domains is intrinsically dependent on their selectivity for specific membrane components; however, while the physicochemical determinants of raft association for lipids are known, very few systematic studies have focused on the structural aspects that guide raft partitioning of proteins. In this review, we describe biophysical and thermodynamic aspects of raft-mimetic liquid ordered phases, focusing on those most relevant for protein partitioning. Further, we detail the variety of experimental models used to study protein-raft interactions. Finally, we review the existing literature on mechanisms for raft targeting, including lipid post-translational modifications, lipid binding, and transmembrane domain features. We conclude that while protein palmitoylation is a clear raft-targeting signal, few other general structural determinants for raft partitioning have been revealed, suggesting that many discoveries lie ahead in this burgeoning field.

  3. Beneath the Minerals, a Layer of Round Lipid Particles Was Identified to Mediate Collagen Calcification in Compact Bone Formation

    PubMed Central

    Xu, Shaohua; Yu, Jianqing J.

    2006-01-01

    Astronauts lose 1–2% of their bone minerals per month during space flights. A systematic search for a countermeasure relies on a good understanding of the mechanism of bone formation at the molecular level. How collagen fibers, the dominant matrix protein in bones, are mineralized remains mysterious. Atomic force microscopy was carried out, in combination with immunostaining and Western blotting, on bovine tibia to identify unrecognized building blocks involved in bone formation and for an elucidation of the process of collagen calcification in bone formation. Before demineralization, tiles of hydroxyapatite crystals were found stacked along bundles of collagen fibers. These tiles were homogeneous in size and shape with dimensions 0.69 × 0.77 × 0.2 μm3. Demineralization dissolved these tiles and revealed small spheres with an apparent diameter around 145 nm. These spheres appeared to be lipid particles since organic solvents dissolved them. The parallel collagen bundles had widths mostly <2 μm. Composition analysis of compact bones indicated a high content of apolar lipids, including triglycerides and cholesterol esters. Apolar lipids are known to form lipid droplets or lipoproteins, and these spheres are unlikely to be matrix vesicles as reported for collagen calcification in epiphyseal cartilages. Results from this study suggest that the layer of round lipid particles on collagen fibers mediates the mineral deposition onto the fibers. The homogeneous size of these lipid particles and the presence of apolipoprotein in demineralized bone tissue suggest the possibility that these particles might be of lipoprotein origin. More studies are needed to verify the last claim and to exclude the possibility that they are secreted lipid droplets. PMID:16980361

  4. Direct comparison of fatty acid ratios in single cellular lipid droplets as determined by Raman spectroscopy and gas chromatography

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cellular lipid droplets are the least studied and least understood cellular organelles in eukaryotic and prokaryotic cells. Despite a broad research trying to understand lipid droplets it has not been possible to determine the composition of individual cellular lipid droplets. In this paper we prese...

  5. Some contributions to the thin-layer chromatographic analysis of complex natural phospholipid and neutral lipid mixtures.

    PubMed

    Helmy, F M; Hack, M H

    1986-01-10

    We have isolated a minor phosphatidyl ethanolamine component from pancreas and a minor phosphatidyl choline component from retina, which were revealed by their separate thin-layer chromatographic properties on silica gel and aluminum oxide sheets, respectively. We have described in some detail a number of modifications in thin-layer chromatography methodology which enhances the opportunity for assessing the glycerophospholipid and neutral lipid composition of tissues, as attested by a diverse set of examples, and have pointed out some of the associated technical problems.

  6. Thin-layer chromatographic analysis of myelin lipids, their differential O-deacylation by primary alkylamines and their selective staining by thionine. A limited phylogenetic study.

    PubMed

    Hack, M H; Helmy, F M

    1990-02-23

    A silica gel thin-layer chromatographic procedure is described for the study of the myelin lipid patterns in a small phylogenetic series of nerve tissue specimens. It involves the selective staining by the thiazine dye thionine and the interpretations were facilitated by a preceding primary alkylamine O-deacylation step. Glycolipids, including sulfatides, and ethanolamine plasmalogens were the principal characterizing lipids.

  7. Immobilization of Lipid Substrates: Application on Phospholipase A2 Determination.

    PubMed

    Karkabounas, Athanassios; Georgiadou, Dimitra G; Argitis, Panagiotis; Psycharis, Vassilios; Nakos, George; Kosmas, Agni M; Lekka, Marilena E

    2015-12-01

    The purpose of the study was to assess a fluorimetric assay for the determination of total phospholipase A(2) (PLA(2)) activity in biological samples introducing the innovation of immobilized substrates on crosslinked polymeric membranes. The immobilized C(12)-NBD-PtdCho, a fluorescent analogue of phosphatidylcholine, exhibited excellent stability for 3 months at 4 °C and was not desorbed in the aqueous reaction mixture during analysis. The limit of detection was 0.5 pmol FA (0.2 pg) and the linear part of the response curve extended from 1 up to 190 nmol FA/h/mL sample. The intra- and inter-day relative standard deviations (%RSD), were ≤6 and ≤9 %, respectively. Statistical comparison with other fluorescent methods showed excellent correlation and agreement. Semiempirical calculations showed a fair amount of electrostatic interaction between the NBD-labeled substrate and the crosslinked polyvinyl alcohol with the styryl pyridinium residues (PVA-SbQ) material, from the plane of which, the sn-2 acyl chain of the phospholipid stands out and is accessible by PLA(2). Atomic Force Microscopy revealed morphological alterations of the immobilized substrate after the reaction with PLA(2). Mass spectrometry showed that only C(12)-NBD-FA, the PLA(2 )hydrolysis product, was detected in the reaction mixture, indicating that PLA(2) recognizes PVA-SbQ/C(12)-NBD-PtdCho as a surface to perform catalysis. PMID:26449236

  8. Experimental Determination of High-Order Bending Elastic Constants of Lipid Bilayers.

    PubMed

    Toscano-Flores, Liliana G; Jacinto-Méndez, Damián; Carbajal-Tinoco, Mauricio D

    2016-06-30

    We present a method to describe the formation of small lipid vesicles in terms of three bending elastic constants that can be experimentally measured. Our method combines a general expression of the elastic free energy of the bilayer and the thermodynamic description of molecular aggregation. The resulting model requires the size distribution of liposomes, which is determined from the X-ray scattered intensity spectra of vesicular dispersions. By using two different preparation methods, we studied a series of vesicular solutions made of distinct lipids and we obtained their corresponding bending elastic constants that are consistent with known bending rigidities. PMID:27267752

  9. Improved pharmacokinetics and enhanced tumor growth inhibition using a nanostructured lipid carrier loaded with doxorubicin and modified with a layer-by-layer polyelectrolyte coating.

    PubMed

    Mussi, Samuel V; Parekh, Gaurav; Pattekari, Pravin; Levchenko, Tatyana; Lvov, Yuri; Ferreira, Lucas A M; Torchilin, Vladimir P

    2015-11-10

    A nanostructured lipid carrier (NLC) loaded with doxorubicin (DOX) has been shown to be cytotoxic against the human cancer cell lines A549 and MCF-7/Adr. In attempts to improve formulation characteristics, enhance pharmacokinetics and antitumor effects, we modified the surface of these NLC with an alternating layer-by-layer (LbL) assembly of polycation and polyanion polyelectrolytes and an additional coating with PEG using a simple method of core shell attachment. The formulation had a narrow size distribution, longer residence in the blood, lower accumulation in the liver, higher accumulation in tumors and a significant tumor growth inhibition effect. Thus, NLC-DOX nanopreparations complexes modified by LbL coating have the potential to enhance the anticancer effects of DOX against tumors. PMID:26325314

  10. Change in Tear Film Lipid Layer Thickness, Corneal Thickness, Volume and Topography after Superficial Cauterization for Conjunctivochalasis

    PubMed Central

    Chan, Tommy C. Y.; Ye, Cong; Ng, Paul KF; Li, Emmy Y. M.; Yuen, Hunter K. L.; Jhanji, Vishal

    2015-01-01

    We evaluated the change in tear film lipid layer thickness, corneal thickness, volume and topography after superficial cauterization of symptomatic conjunctivochalasis. Bilateral superficial conjunctival cauterization was performed in 36 eyes of 18 patients with symptomatic conjunctivochalasis. The mean age of patients (12 males, 6 females) was 68.6 ± 10.9 years (range: 44–83 years). Preoperatively, 28 eyes (77.8%) had grade 1 conjunctivochalasis, and 8 eyes (22.2%) had grade 2 conjunctivochalasis. At 1 month postoperatively, the severity of conjunctivochalasis decreased significantly (p < 0.001) and 29 eyes (80.6%) had grade 0 conjunctivochalasis whereas 7 eyes (19.4%) had grade 1 conjunctivochalasis. The mean Ocular Surface Disease Index score decreased from 31.5 ± 15.2 preoperatively to 21.5 ± 14.2 at the end of 1 month postoperatively (p = 0.001). There was a statistically significant increase in mean tear film lipid layer thickness 1 month after the surgery (49.6 ± 16.1 nm vs 62.6 ± 21.6 nm; p < 0.001). The central corneal thickness, thinnest corneal thickness and corneal volume decreased significantly postoperatively (p < 0.001). Our study showed that superficial conjunctival cauterization is an effective technique for management of conjunctivochalasis in the short term. An increase in tear film lipid layer thickness along with a decrease in corneal thickness and volume were observed after surgical correction of conjunctivochalasis. PMID:26184418

  11. Change in Tear Film Lipid Layer Thickness, Corneal Thickness, Volume and Topography after Superficial Cauterization for Conjunctivochalasis.

    PubMed

    Chan, Tommy C Y; Ye, Cong; Ng, Paul K F; Li, Emmy Y M; Yuen, Hunter K L; Jhanji, Vishal

    2015-01-01

    We evaluated the change in tear film lipid layer thickness, corneal thickness, volume and topography after superficial cauterization of symptomatic conjunctivochalasis. Bilateral superficial conjunctival cauterization was performed in 36 eyes of 18 patients with symptomatic conjunctivochalasis. The mean age of patients (12 males, 6 females) was 68.6 ± 10.9 years (range: 44-83 years). Preoperatively, 28 eyes (77.8%) had grade 1 conjunctivochalasis, and 8 eyes (22.2%) had grade 2 conjunctivochalasis. At 1 month postoperatively, the severity of conjunctivochalasis decreased significantly (p < 0.001) and 29 eyes (80.6%) had grade 0 conjunctivochalasis whereas 7 eyes (19.4%) had grade 1 conjunctivochalasis. The mean Ocular Surface Disease Index score decreased from 31.5 ± 15.2 preoperatively to 21.5 ± 14.2 at the end of 1 month postoperatively (p = 0.001). There was a statistically significant increase in mean tear film lipid layer thickness 1 month after the surgery (49.6 ± 16.1 nm vs 62.6 ± 21.6 nm; p < 0.001). The central corneal thickness, thinnest corneal thickness and corneal volume decreased significantly postoperatively (p < 0.001). Our study showed that superficial conjunctival cauterization is an effective technique for management of conjunctivochalasis in the short term. An increase in tear film lipid layer thickness along with a decrease in corneal thickness and volume were observed after surgical correction of conjunctivochalasis. PMID:26184418

  12. Direct comparison of fatty acid ratios in single cellular lipid droplets as determined by comparative Raman spectroscopy and gas chromatography.

    PubMed

    Schie, Iwan W; Nolte, Lena; Pedersen, Theresa L; Smith, Zach; Wu, Jian; Yahiatène, Idir; Newman, John W; Huser, Thomas

    2013-11-01

    Cellular lipid droplets are the least studied and least understood cellular organelles in eukaryotic and prokaryotic cells. Despite a significant body of research studying the physiology of lipid droplets it has not yet been possible to fully determine the composition of individual cellular lipid droplets. In this paper we use Raman spectroscopy on single cellular lipid droplets and least-squares fitting of pure fatty acid spectra to determine the composition of individual lipid droplets in cells after treatment with different ratios of oleic and palmitic acid. We validate the results of the Raman spectroscopy-based single lipid droplet analysis with results obtained by gas chromatography analysis of millions of cells, and find that our approach can accurately predict the relative amount of a specific fatty acid in the lipid droplet. Based on these results we show that the fatty acid composition in individual lipid droplets is on average similar to that of all lipid droplets found in the sample. Furthermore, we expand this approach to the investigation of the lipid composition in single cellular peroxisomes. We determine the location of cellular peroxisomes based on two-photon excitation fluorescence (TPEF) imaging of peroxisomes labeled with the green fluorescent protein, and successive Raman spectroscopy of peroxisomes. We find that in some cases peroxisomes can produce a detectable CARS signal, and that the peroxisomal Raman spectra exhibit an oleic acid-like signature.

  13. Nile red fluorescence screening facilitating neutral lipid phenotype determination in budding yeast, Saccharomyces cerevisiae, and the fission yeast Schizosaccharomyces pombe.

    PubMed

    Rostron, Kerry A; Rolph, Carole E; Lawrence, Clare L

    2015-07-01

    Investigation of yeast neutral lipid accumulation is important for biotechnology and also for modelling aberrant lipid metabolism in human disease. The Nile red (NR) method has been extensively utilised to determine lipid phenotypes of yeast cells via microscopic means. NR assays have been used to differentiate lipid accumulation and relative amounts of lipid in oleaginous species but have not been thoroughly validated for phenotype determination arising from genetic modification. A modified NR assay, first described by Sitepu et al. (J Microbiol Methods 91:321-328, 2012), was able to detect neutral lipid changes in Saccharomyces cerevisiae deletion mutants with sensitivity similar to more advanced methodology. We have also be able to, for the first time, successfully apply the NR assay to the well characterised fission yeast Schizosaccharomyces pombe, an increasingly important organism in biotechnology. The described NR fluorescence assay is suitable for increased throughput and rapid screening of genetically modified strains in both the biotechnology industry and for modelling ectopic lipid production for a variety of human diseases. This ultimately negates the need for labour intensive and time consuming lipid analyses of samples that may not yield a desirable lipid phenotype, whilst genetic modifications impacting significantly on the cellular lipid phenotype can be further promoted for more in depth analyses.

  14. Determination of lipid content of oleaginous microalgal biomass by NMR spectroscopic and GC-MS techniques.

    PubMed

    Sarpal, Amarijt S; Teixeira, Claudia M L L; Silva, Paulo R M; Lima, Gustavo M; Silva, Samantha R; Monteiro, Thays V; Cunha, Valnei S; Daroda, Romeu J

    2015-05-01

    Direct methods based on (1)H NMR spectroscopic techniques have been developed for the determination of neutral lipids (triglycerides and free fatty acids) and polar lipids (glyceroglycolipids/phospholipids) in the solvent extracts of oleaginous microalgal biomasses cultivated on a laboratory scale with two species in different media. The chemical shift assignments observed in the (1)H and (13)C NMR spectra corresponding to unsaturated (C18:N, N = 1-3, C20:3, C20:5, C22:6, epoxy) and saturated (C14-C18) fatty acid ester components in a complex matrix involving overlapped resonances have been unambiguously confirmed by the application of 2D NMR spectroscopy (total correlation spectroscopy and heteronuclear single quantum coherence-total correlation spectroscopy). The study of the effect of a polar lipid matrix on the determination of neutral lipids by an internal reference blending process by a systematic designed experimental protocol has provided absolute quantification. The fatty acid composition of algal extracts was found to be similar to that of vegetable oils containing saturated (C16-C18:0) and unsaturated (C18:N, N = 1-3, C20:N, N = 3-4, C22:6) fatty acids as confirmed by NMR spectroscopy and gas chromatography-mass spectrometry analyses. The NMR methods developed offer great potential for rapid screening of algal strains for generation of algal biomass with the desired lipid content, quality, and potential for biodiesel and value-added polyunsaturated fatty acids in view of the cost economics of the overall cost of generation of the biomass. PMID:25801382

  15. Determination of PCBs and total lipids in edible fish and crab tissue using supercritical fluid extraction

    SciTech Connect

    Gavlor, M.; Hale, R.; Smith, C.; Thames, J.; Mothershead, R.

    1995-12-31

    An offline supercritical fluid extraction (SFE) method has been developed to determine PCB congeners and total tissue lipid content in edible fish and crab tissues collected from several river systems in Virginia. The method is rapid and safe, requiring only 40 minutes per sample and uses nonorganic solvents for total lipid extraction and only 1.5 mL isooctane for PCB extraction. The SFE approach compares favorably with soxhlet extraction, ASE and column elution. Over 800 fish and crab tissue samples were analyzed successfully, thus demonstrating the robustness of the method. Total lipid values obtained using SFE showed considerable spatial and interspecies variability ranging from 1.8% in blue crab (Callinectes sapidus) to 36.4% in striped bass (Morone saxatilis). Total PCB concentrations also varied greatly by site and species. These ranged from below the quantitation limit (1.0 {micro}1 g/kg) to 9,910 {micro}g/kg on a dry weight basis using GCELCD. Dominant PCB congeners detected were in good agreement with those reported by other researchers. Mean total PCB concentrations did not correlate well with total tissue lipid content.

  16. The effect of supplementing layer diets with shark cartilage or chitosan on egg components and yolk lipids.

    PubMed

    Nogueira, C M; Zapata, J F F; Fuentes, M F F; Freitas, E R; Craveiro, A A; Aguiar, C M

    2003-05-01

    1. An experiment was designed to evaluate the effects of the addition of shark cartilage (SC) or chitosan (CH) to layer diets on egg component weights, yolk lipids and hen plasma lipids. 2. Hy-Line laying hens (80) were used during a 56 d feeding trial. Treatments were: basal diet (BD), BD + 20 g/kg SC, BD + 30 g/kg SC, BD + 20 g/kg CH and BD + 30 g/kg CH. Eggs were analysed on d 14, 28, 42 and 56. 3. Egg weight and egg component weights were not affected by these treatments throughout the experimental period. 4. After 14d of experimental feeding, cholesterol levels were higher in eggs from birds given BD + 20 g/kg CH and BD + 30 g/kg CH than in those from birds given BD. 5. Furthermore, eggs from hens given BD + 20 g/kg SC or BD + 20 g/kg CH were higher in palmitic and stearic acids and lower in oleic acid than those from birds fed on BD. After 56 d feeding, however, palmitic and stearic acid contents in eggs from hens given any of the supplemented diets were lower than in those from hens given BD, and oleic acid in eggs from hens given BD + 20 g/kg SC, BD + 30 g/kg SC and BD + 30 g/kg CH was higher than in those from birds fed on BD. 6. Plasma cholesterol and triacylglycerol levels were not significantly affected by dietary treatment. 7. Shark cartilage or chitosan at up to 30 g/kg in layer diets did not affect egg component weights (yolk, white and shell) and total lipid contents. During the period from 42 to 56d of experimental feeding, diets containing up to 30 g/kg chitosan reduced egg yolk contents of cholesterol, palmitic and stearic acids and increased the content of oleic acid. PMID:12828207

  17. Development of the layer-by-layer biosensor using graphene films: application for cholesterol determination

    NASA Astrophysics Data System (ADS)

    Binh Nguyen, Hai; Chuc Nguyen, Van; Nguyen, Van Tu; Doan Le, Huu; Quynh Nguyen, Van; Thanh Tam Ngo, Thi; Phuc Do, Quan; Nghia Nguyen, Xuan; Phan, Ngoc Minh; Tran, Dai Lam

    2013-03-01

    The preparation and characterization of graphene films for cholesterol determination are described. The graphene films were synthesized by thermal chemical vapor deposition (CVD) method. Methane gas (CH4) and copper tape were used as carbon source and catalyst in the graphene growth process, respectively. The intergrated array was fabricated by using micro-electro-mechanical systems (MEMS) technology in which Fe3O4-doped polyaniline (PANi) film was electropolymerized on Pt/Gr electrodes. The properties of the Pt/Gr/PANi/Fe3O4 films were investigated by field-emission scanning electron microscopy (FE-SEM), Raman spectroscopy and electrochemical techniques. Cholesterol oxidase (ChOx) has been immobilized onto the working electrode with glutaraldehyde agent. The cholesterol electrochemical biosensor shows high sensitivity (74 μA mM-1 cm-2) and fast response time (<5 s). A linear calibration plot was obtained in the wide cholesterol concentration range from 2 to 20 mM and correlation coefficient square (R2) of 0.9986. This new layer-by-layer biosensor based on graphene films promises many practical applications.

  18. Determination of layer-charge characteristics of smectites

    USGS Publications Warehouse

    Christidis, G.E.; Eberl, D.D.

    2003-01-01

    A new method for calculation of layer charge and charge distribution of smectites is proposed. The method is based on comparisons between X-ray diffraction (XRD) patterns of K-saturated, ethylene glycol-solvated, oriented samples and calculated XRD patterns for three-component, mixed-layer systems. For the calculated patterns it is assumed that the measured patterns can be modeled as random interstratifications of fully expanding 17.1 A?? layers, partially expanding 13.5 A?? layers and non-expanding 9.98 A?? layers. The technique was tested using 29 well characterized smectites. According to their XRD patterns, smectites were classified as group 1 (low-charge smectites) and group 2 (high-charge smectites). The boundary between the two groups is at a layer charge of -0.46 equivalents per half unit-cell. Low-charge smectites are dominated by 17.1 A?? layers, whereas high-charge smectites contain only 20% fully expandable layers on average. Smectite properties and industrial applications may be dictated by the proportion of 17.1 A?? layers present. Non-expanding layers may control the behavior of smectites during weathering, facilitating the formation of illite layers after subsequent cycles of wetting and drying. The precision of the method is better than 3.5% at a layer charge of -0.50; therefore the method should be useful for basic research and for industrial purposes.

  19. Experimental determination of wave function spread in Si inversion layers

    NASA Astrophysics Data System (ADS)

    Majumdar, Amlan

    2010-08-01

    We have experimentally determined the extent of wave function spread TQM in Si inversion layers on (100)-oriented surface in metal-oxide-semiconductor field-effect transistors (MOSFETs) using the back gate bias sensitivity of front gate threshold voltage of planar fully depleted silicon-on-insulator (SOI) MOSFETs. We show that the sum of TQM for large positive and negative F is an electrically determined value of the SOI thickness TSI. We find that the electric field dependence of TQM for electrons and holes is given by TQM˜F-0.4 and F-0.6, respectively, at high electric fields with TQM being larger for holes at a given F. Larger TQM for holes can be explained by the fact that holes have a smaller effective mass along the confinement direction than electrons in (100) Si. The field dependences of TQM are, however, not consistent with the results of variational calculations that assume single-subband occupancy and predict TQM˜F-1/3. The discrepancy likely indicates that the effects of multiple-subband occupation are significant at room temperature, especially for holes.

  20. Application of solid/liquid extraction for the gravimetric determination of lipids in royal jelly.

    PubMed

    Antinelli, Jean-François; Davico, Renée; Rognone, Catherine; Faucon, Jean-Paul; Lizzani-Cuvelier, Louisette

    2002-04-10

    Gravimetric lipid determination is a major parameter for the characterization and the authentication of royal jelly quality. A solid/liquid extraction was compared to the reference method, which is based on liquid/liquid extraction. The amount of royal jelly and the time of the extraction were optimized in comparison to the reference method. Boiling/rinsing ratio and spread of royal jelly onto the extraction thimble were identified as critical parameters, resulting in good accuracy and precision for the alternative method. Comparison of reproducibility and repeatability of both methods associated with gas chromatographic analysis of the composition of the extracted lipids showed no differences between the two methods. As the intra-laboratory validation tests were comparable to the reference method, while offering rapidity and a decrease in amount of solvent used, it was concluded that the proposed method should be used with no modification of quality criteria and norms established for royal jelly characterization.

  1. Altered lipid metabolism in the aging kidney identified by three layered omic analysis.

    PubMed

    Braun, Fabian; Rinschen, Markus M; Bartels, Valerie; Frommolt, Peter; Habermann, Bianca; Hoeijmakers, Jan H J; Schumacher, Björn; Dollé, Martijn E T; Müller, Roman-Ulrich; Benzing, Thomas; Schermer, Bernhard; Kurschat, Christine E

    2016-03-01

    Aging-associated diseases and their comorbidities affect the life of a constantly growing proportion of the population in developed countries. At the center of these comorbidities are changes of kidney structure and function as age-related chronic kidney disease predisposes to the development of cardiovascular diseases such as stroke, myocardial infarction or heart failure. To detect molecular mechanisms involved in kidney aging, we analyzed gene expression profiles of kidneys from adult and aged wild-type mice by transcriptomic, proteomic and targeted lipidomic methodologies. Interestingly, transcriptome and proteome analyses revealed differential expression of genes primarily involved in lipid metabolism and immune response. Additional lipidomic analyses uncovered significant age-related differences in the total amount of phosphatidylethanolamines, phosphatidylcholines and sphingomyelins as well as in subspecies of phosphatidylserines and ceramides with age. By integration of these datasets we identified Aldh1a1, a key enzyme in vitamin A metabolism specifically expressed in the medullary ascending limb, as one of the most prominent upregulated proteins in old kidneys. Moreover, ceramidase Asah1 was highly expressed in aged kidneys, consistent with a decrease in ceramide C16. In summary, our data suggest that changes in lipid metabolism are involved in the process of kidney aging and in the development of chronic kidney disease. PMID:26886165

  2. Altered lipid metabolism in the aging kidney identified by three layered omic analysis

    PubMed Central

    Braun, Fabian; Rinschen, Markus M.; Bartels, Valerie; Frommolt, Peter; Habermann, Bianca; Hoeijmakers, Jan H.J.; Schumacher, Björn; Dollé, Martijn E.T.; Müller, Roman-Ulrich; Benzing, Thomas; Schermer, Bernhard; Kurschat, Christine E.

    2016-01-01

    Aging-associated diseases and their comorbidities affect the life of a constantly growing proportion of the population in developed countries. At the center of these comorbidities are changes of kidney structure and function as age-related chronic kidney disease predisposes to the development of cardiovascular diseases such as stroke, myocardial infarction or heart failure. To detect molecular mechanisms involved in kidney aging, we analyzed gene expression profiles of kidneys from adult and aged wild-type mice by transcriptomic, proteomic and targeted lipidomic methodologies. Interestingly, transcriptome and proteome analyses revealed differential expression of genes primarily involved in lipid metabolism and immune response. Additional lipidomic analyses uncovered significant age-related differences in the total amount of phosphatidylethanolamines, phosphatidylcholines and sphingomyelins as well as in subspecies of phosphatidylserines and ceramides with age. By integration of these datasets we identified Aldh1a1, a key enzyme in vitamin A metabolism specifically expressed in the medullary ascending limb, as one of the most prominent upregulated proteins in old kidneys. Moreover, ceramidase Asah1 was highly expressed in aged kidneys, consistent with a decrease in ceramide C16. In summary, our data suggest that changes in lipid metabolism are involved in the process of kidney aging and in the development of chronic kidney disease. PMID:26886165

  3. Effect of cholesterol on the phase behavior of solid-supported lipid vesicle layers.

    PubMed

    Losada-Pérez, P; Khorshid, M; Yongabi, D; Wagner, P

    2015-04-16

    The interest in solid-supported biomimetic membranes stems from their utility in nanotechnology and biosensing. In particular, supported lipid vesicles (SLVs) have become popular in both fundamental biophysical studies and pharmaceutical screening applications. It is thus essential to gain information on the structural properties and phase behavior of SLVs. Here we report on a study on the influence of cholesterol on the phase behavior of SLVs of saturated phospholipids by using quartz crystal microbalance with dissipation monitoring, a label-free and nonintrusive surface-sensitive technique. Two complementary approaches have been used, a Voigt-based viscoelastic model yielding shear viscosity temperature profiles and the first-order derivative of the frequency (mass-sensitive) shifts. Anomalies in the shear viscosity and extrema in the first-order derivative frequency curves stand as a token of the main phase transition and provide information on its gradual suppression upon addition of cholesterol. This method proves convenient for its small sample volume needed, its short temperature equilibration time and the non-necessity of external labels. This work can be regarded as a starting point for further studies on more rare lipid systems and different geometries, such as tethered SLVs or biologically relevant vesicles produced by living cells.

  4. O3-induced formation of bioactive lipids: estimated surface concentrations and lining layer effects.

    PubMed

    Postlethwait, E M; Cueto, R; Velsor, L W; Pryor, W A

    1998-06-01

    Recent evidence suggests that inhaled ozone (O3) does not induce toxicity via direct epithelial interactions. Reactions with epithelial lining fluid (ELF) constituents limit cellular contact and generate products, including lipid ozonation products, postulated to initiate pathophysiological cascades. To delineate specific aspects of lipid ozonation product formation and to estimate in situ surface concentrations, we studied the O3 absorption characteristics of ELF constituent mixtures and measured hexanal, heptanal, and nonanal yields as a function of ascorbic acid (AH2) concentration. Exposures of isolated rat lungs, bronchoalveolar lavage fluid (BALF) and egg phosphatidylcholine (PC) liposomes were conducted. 1) O3 absorption by AH2, uric acid, and albumin exceeded that by egg PC and glutathione. O3 reaction with egg PC occurred when AH2 concentrations were reduced. 2) Aldehydes were produced in low yield during lung and BALF exposures in a time- and O3 concentration-dependent manner. 3) Diminishing BALF AH2 content lowered O3 uptake but increased aldehyde yields. Conversely, AH2 addition to egg PC increased O3 uptake but reduced aldehyde yields. Estimations of bioactive ozonation and autoxidation product accumulation within the ELF suggested possible nanomolar to low micromolar concentrations. The use of reaction products as metrics of O3 exposure may have intrinsic sensitivity and specificity limitations. Moreover, due to the heterogenous nature of O3 reactions within the ELF, dose-response relationships may not be linear with respect to O3 absorption.

  5. Quantitative thin layer chromatography for the analysis of skin surface lipids. A time-saving method using a new TLC plate.

    PubMed

    Weissmann, A

    1979-07-30

    Recently a new thin layer chromatography plate (Whatman LK 6D) became available which is extremely easy to handle and permits highly reproducible qualitative and quantitative analysis. This plate proved to be of great value for the investigation of skin surface lipids. The use of a fatty acid methyl ester as an internal standard makes it unnecessary to employ additional gravimetrical or photometrical methods for quanitative lipid analysis. The method presented in this paper is simpler and requires much less time than alternative procedures and allows a large number of lipid samples to be processed simultaneously. PMID:475450

  6. Interplay of electrostatics and lipid packing determines the binding of charged polymer coated nanoparticles to model membranes.

    PubMed

    Biswas, Nupur; Bhattacharya, Rupak; Saha, Arindam; Jana, Nikhil R; Basu, Jaydeep K

    2015-10-01

    Understanding of nanoparticle-membrane interactions is useful for various applications of nanoparticles like drug delivery and imaging. Here we report on the studies of interaction between hydrophilic charged polymer coated semiconductor quantum dot nanoparticles with model lipid membranes. Atomic force microscopy and X-ray reflectivity measurements suggest that cationic nanoparticles bind and penetrate bilayers of zwitterionic lipids. Penetration and binding depend on the extent of lipid packing and result in the disruption of the lipid bilayer accompanied by enhanced lipid diffusion. On the other hand, anionic nanoparticles show minimal membrane binding although, curiously, their interaction leads to reduction in lipid diffusivity. It is suggested that the enhanced binding of cationic QDs at higher lipid packing can be understood in terms of the effective surface potential of the bilayers which is tunable through membrane lipid packing. Our results bring forth the subtle interplay of membrane lipid packing and electrostatics which determine nanoparticle binding and penetration of model membranes with further implications for real cell membranes.

  7. The Transmembrane Helix Tilt May Be Determined by the Balance between Precession Entropy and Lipid Perturbation

    PubMed Central

    2012-01-01

    Hydrophobic helical peptides interact with lipid bilayers in various modes, determined by the match between the length of the helix’s hydrophobic core and the thickness of the hydrocarbon region of the bilayer. For example, long helices may tilt with respect to the membrane normal to bury their hydrophobic cores in the membrane, and the lipid bilayer may stretch to match the helix length. Recent molecular dynamics simulations and potential of mean force calculations have shown that some TM helices whose lengths are equal to, or even shorter than, the bilayer thickness may also tilt. The tilt is driven by a gain in the helix precession entropy, which compensates for the free energy penalty resulting from membrane deformation. Using this free energy balance, we derived theoretically an equation of state, describing the dependence of the tilt on the helix length and membrane thickness. To this end, we conducted coarse-grained Monte Carlo simulations of the interaction of helices of various lengths with lipid bilayers of various thicknesses, reproducing and expanding the previous molecular dynamics simulations. Insight from the simulations facilitated the derivation of the theoretical model. The tilt angles calculated using the theoretical model agree well with our simulations and with previous calculations and measurements. PMID:24932138

  8. Virosome engineering of colloidal particles and surfaces: bioinspired fusion to supported lipid layers.

    PubMed

    Fleddermann, J; Diamanti, E; Azinas, S; Košutić, M; Dähne, L; Estrela-Lopis, I; Amacker, M; Donath, E; Moya, S E

    2016-04-21

    Immunostimulating reconstituted influenza virosomes (IRIVs) are liposomes with functional viral envelope glycoproteins: influenza virus hemagglutinin (HA) and neuraminidase intercalated in the phospholipid bilayer. Here we address the fusion of IRIVs to artificial supported lipid membranes assembled on polyelectrolyte multilayers on both colloidal particles and planar substrates. The R18 assay is used to prove the IRIV fusion in dependence of pH, temperature and HA concentration. IRIVs display a pH-dependent fusion mechanism, fusing at low pH in analogy to the influenza virus. The pH dependence is confirmed by the Quartz Crystal Microbalance technique. Atomic Force Microscopy imaging shows that at low pH virosomes are integrated in the supported membrane displaying flattened features and a reduced vertical thickness. Virosome fusion offers a new strategy for transferring biological functions on artificial supported membranes with potential applications in targeted delivery and sensing. PMID:27006101

  9. Super-resolved thickness maps of thin film phantoms and in vivo visualization of tear film lipid layer using OCT

    PubMed Central

    dos Santos, Valentin Aranha; Schmetterer, Leopold; Triggs, Graham J.; Leitgeb, Rainer A.; Gröschl, Martin; Messner, Alina; Schmidl, Doreen; Garhofer, Gerhard; Aschinger, Gerold; Werkmeister, René M.

    2016-01-01

    In optical coherence tomography (OCT), the axial resolution is directly linked to the coherence length of the employed light source. It is currently unclear if OCT allows measuring thicknesses below its axial resolution value. To investigate spectral-domain OCT imaging in the super-resolution regime, we derived a signal model and compared it with the experiment. Several island thin film samples of known refractive indices and thicknesses in the range 46 – 163 nm were fabricated and imaged. Reference thickness measurements were performed using a commercial atomic force microscope. In vivo measurements of the tear film were performed in 4 healthy subjects. Our results show that quantitative super-resolved thickness measurement can be performed using OCT. In addition, we report repeatable tear film lipid layer visualization. Our results provide a novel interpretation of the OCT axial resolution limit and open a perspective to deeper extraction of the information hidden in the coherence volume. PMID:27446696

  10. Virosome engineering of colloidal particles and surfaces: bioinspired fusion to supported lipid layers

    NASA Astrophysics Data System (ADS)

    Fleddermann, J.; Diamanti, E.; Azinas, S.; Košutić, M.; Dähne, L.; Estrela-Lopis, I.; Amacker, M.; Donath, E.; Moya, S. E.

    2016-04-01

    Immunostimulating reconstituted influenza virosomes (IRIVs) are liposomes with functional viral envelope glycoproteins: influenza virus hemagglutinin (HA) and neuraminidase intercalated in the phospholipid bilayer. Here we address the fusion of IRIVs to artificial supported lipid membranes assembled on polyelectrolyte multilayers on both colloidal particles and planar substrates. The R18 assay is used to prove the IRIV fusion in dependence of pH, temperature and HA concentration. IRIVs display a pH-dependent fusion mechanism, fusing at low pH in analogy to the influenza virus. The pH dependence is confirmed by the Quartz Crystal Microbalance technique. Atomic Force Microscopy imaging shows that at low pH virosomes are integrated in the supported membrane displaying flattened features and a reduced vertical thickness. Virosome fusion offers a new strategy for transferring biological functions on artificial supported membranes with potential applications in targeted delivery and sensing.Immunostimulating reconstituted influenza virosomes (IRIVs) are liposomes with functional viral envelope glycoproteins: influenza virus hemagglutinin (HA) and neuraminidase intercalated in the phospholipid bilayer. Here we address the fusion of IRIVs to artificial supported lipid membranes assembled on polyelectrolyte multilayers on both colloidal particles and planar substrates. The R18 assay is used to prove the IRIV fusion in dependence of pH, temperature and HA concentration. IRIVs display a pH-dependent fusion mechanism, fusing at low pH in analogy to the influenza virus. The pH dependence is confirmed by the Quartz Crystal Microbalance technique. Atomic Force Microscopy imaging shows that at low pH virosomes are integrated in the supported membrane displaying flattened features and a reduced vertical thickness. Virosome fusion offers a new strategy for transferring biological functions on artificial supported membranes with potential applications in targeted delivery and sensing

  11. Evaluation of FT-IR and Nile Red methods for microalgal lipid characterization and biomass composition determination.

    PubMed

    Feng, Guo-Dong; Zhang, Fang; Cheng, Li-Hua; Xu, Xin-Hua; Zhang, Lin; Chen, Huan-Lin

    2013-01-01

    To characterize lipid content of microalgal cells rapidly and accurately, the gravimetric determination, FT-IR and Nile Red (NR) staining were investigated on six typical eukaryotic and prokaryotic algae species. FT-IR and Nile Red were relative quantification methods and a standard curve was required in contrast to the gravimetric method. The FT-IR method determined the lipid, carbohydrate and protein contents simultaneously assuming that the algal cells only consisted of those three components. The Nile Red method was a relatively rapid method for neutral lipid content characterization by spectrofluorometry and could locate lipid body of the algal cell by fluorescence microscopy. According to sample sources and processing purposes, the gravimetric determination was preferable for large-scale cultivation with low-frequency monitoring, while FT-IR and Nile Red were suitable for general laboratory cultivation with medium-frequency monitoring, in particularly Nile Red was appropriate for small samples when high-frequency screening was required. PMID:23196229

  12. The Effect of Particle Size on the Deposition of Solid Lipid Nanoparticles in Different Skin Layers: A Histological Study

    PubMed Central

    Mardhiah Adib, Zahra; Ghanbarzadeh, Saeed; Kouhsoltani, Maryam; Yari Khosroshahi, Ahmad; Hamishehkar, Hamed

    2016-01-01

    Purpose: In the present study the effect of particle size, as a substantial parameters in skin penetration, on the deposition depth and rate of SLNs in different layers of skin was explored. Methods: SLNs in different particle size ranges (80, 333 and 971 nm) made of Precirol as solid lipid were prepared using hot melt homogenization technique and pigmented by Rhodamine B to be able to be tracked in the skin under inspection of fluorescent microscopy. After 0.5 h, 3 h, 6 h and 24 h of SLNs administration on rat skin, animals were sacrificed and exercised skins were sliced by a freeze microtome. SLNs were monitored in the skin structure under fluorescence microscope. Results: The size of SLNs played a crucial role in the penetration to deep skin layers. The sub100 nm size range of SLNs showed the most promising skin penetration rate and depth mainly via hair follicles. Conclusion: The results of the present study indicated that the selection of an appropriate size of particles may be a valuable factor impacting the therapeutic outcomes of dermal drug administration. PMID:27123415

  13. Health consequences of postsocialist transition: dietary and lifestyle determinants of plasma lipids in Yakutia.

    PubMed

    Sorensen, M V; Snodgrass, J J; Leonard, W R; Tarskaia, A; Ivanov, K I; Krivoshapkin, V G; Spitsyn, V A

    2005-01-01

    The rapid social and cultural changes introduced by the collapse of the Soviet Union have resulted in important differences in cardiovascular health for indigenous Siberians. This study investigated diet and lifestyle determinants of plasma lipids in the Yakut, an indigenous Siberian herding population. The study used a cross-sectional design with data on 201 subjects in three urbanized towns and three rural communities in northeastern Siberia. Data on sociodemographic characteristics, dietary intake, and material lifestyle were collected, and lipids were analyzed from venous whole blood. Diet was analyzed using patterns of dietary intake based on principal components analysis of a dietary intake (food frequency) questionnaire. We identified three diet patterns: a traditional subsistence diet, a market foods diet, and a mixed diet. The effect of lifestyle on cardiovascular risk factors was measured using an ethnographically defined lifestyle index, with two orthogonal dimensions: subsistence lifestyle and modern lifestyle. Total cholesterol (TC) and low-density lipoprotein (LDL) were significantly higher among those consuming a traditional subsistence diet of meat and dairy products. A modern lifestyle was associated with lower TC and LDL but higher adiposity and higher risk of obesity. LDL and TC were higher in rural communities and lower in urbanized towns. The significantly higher lipid levels associated with a subsistence diet and indirectly with a subsistence lifestyle indicate the emergence of a significant health problem associated with the social and cultural changes occurring in Yakutia today. These findings underscore the need for dietary modification and promotion of physical activity among those most at risk for cardiovascular disease (CVD). Moreover, these results differ from those commonly seen in "modernizing" populations, in that elements of subsistence lifestyle are associated with an elevated rather than reduced risk of CVD. Such variable

  14. Preparation of stabilized magnetic nanoparticles with polymerizable lipid and anchor compounds.

    PubMed

    Kang, Boram; Choi, Suk-Jung

    2014-02-01

    Although the lipid-based method for coating of magnetic nanoparticles (MNPs) is rapid and simple, the unstable state of the lipid layer is a major limitation for the practical application of this method. We devised a method to prepare stabilized MNPs by covalent modifications such as lipid polymerization and anchoring of the lipid layer. The stability of the modified lipid layer was demonstrated by the stable status of enzymes immobilized on the MNPs and the resistance of the MNPs to aggregation. We also determined the maximum ratio of nonpolymerizable lipophilic compounds that can be included in the layer without significantly reducing stability.

  15. Anion determination in damage layers of stone monuments

    NASA Astrophysics Data System (ADS)

    Gobbi, G.; Zappia, G.; Sabbioni, C.

    An analytical procedure was set up for the dissolution of damage layers from stone monuments with the aim of performing ion chromatography analyses. The dissolution was performed using ion exchange resin H-form and the ion chromatography analysis with two serial detectors (conductometer and spectrophotometer). The problem of interferences of different ions is discussed.

  16. Determination of Stability and Translation in a Boundary Layer

    NASA Technical Reports Server (NTRS)

    Crepeau, John; Tobak, Murray

    1996-01-01

    Reducing the infinite degrees of freedom inherent in fluid motion into a manageable number of modes to analyze fluid motion is presented. The concepts behind the center manifold technique are used. Study of the Blasius boundary layer and a precise description of stability within the flow field are discussed.

  17. Formulation and In Vitro and In Vivo Evaluation of Lipid-Based Terbutaline Sulphate Bi-layer Tablets for Once-Daily Administration.

    PubMed

    Hashem, Fahima M; Nasr, Mohamed; Fathy, Gihan; Ismail, Aliaa

    2016-06-01

    The objective of this study was to prepare and evaluate terbutaline sulphate (TBS) bi-layer tablets for once-daily administration. The bi-layer tablets consisted of an immediate-release layer and a sustained-release layer containing 5 and 10 mg TBS, respectively. The sustained-release layer was developed by using Compritol®888 ATO, Precirol® ATO 5, stearic acid, and tristearin, separately, as slowly eroding lipid matrices. A full 4 × 2(2) factorial design was employed for optimization of the sustained-release layer and to explore the effect of lipid type (X 1), drug-lipid ratio (X 2), and filler type (X 3) on the percentage drug released at 8, 12, and 24 h (Y 1, Y 2, and Y 3) as dependent variables. Sixteen TBS sustained-release matrices (F1-F16) were prepared by melt solid dispersion method. None of the prepared matrices achieved the targeted release profile. However, F2 that showed a relatively promising drug release was subjected to trial and error optimization for the filler composition to develop two optimized matrices (F17 and F18). F18 which consisted of drug-Compritol®888 ATO at ratio (1:6 w/w) and Avicel PH 101/dibasic calcium phosphate mixture of 2:1 (w/w) was selected as sustained-release layer. TBS bi-layer tablets were evaluated for their physical properties, in vitro drug release, effect of storage on drug content, and in vivo performance in rabbits. The bi-layer tablets showed acceptable physical properties and release characteristics. In vivo absorption in rabbits revealed initial high TBS plasma levels followed by sustained levels over 24 h compared to immediate-release tablets. PMID:26335420

  18. Diffusion of ionizable solutes across planar lipid bilayer membranes: boundary-layer pH gradients and the effect of buffers.

    PubMed

    Xiang, T X; Anderson, B D

    1993-11-01

    The diffusion of weak acids or bases across planar lipid bilayer membranes results in aqueous boundary layer pH gradients. If not properly taken into account, such pH gradients will lead to errors in estimated membrane permeability coefficients, Pm. The role of the permeant concentration, the buffer capacity, and the physicochemical properties of both permeant and buffer on the magnitude and impact of such pH gradients have been explored. A theoretical model has been developed to describe the diffusion of both permeant and buffer species. Significant pH gradients develop depending on solution pH and the pKa's, concentrations, and Pm values of both permeant and buffer. The relative error in experimentally determined Pm values was calculated as the ratio, r, between apparent Pm values (obtained from flux measurements using an equation which neglected boundary layer pH gradients) and its true value. Simulated r values ranged from 1 (0% error) to < 0.01 (> 100% error) for weak acids, decreasing with decreasing buffer capacity and increasing solute flux. The buffer capacity required for an r > 0.95 was calculated versus pH for permeants varying in pKa and Pm. Membrane-permeable buffers significantly reduce boundary layer pH gradients through a feedback effect due to buffer cotransport. Apparent Pm values of p-hydroxymethyl benzoic acid across lecithin bilayer membranes at 25 degrees C were obtained as a function of permeant concentration in various buffers [glycolic, 2-(N-morpholino)ethane-sulfonic, and formic acids]. Predictions agreed closely with experimental fluxes. PMID:8290481

  19. Consumption of dietary sugar by gut bacteria determines Drosophila lipid content.

    PubMed

    Huang, Jia-Hsin; Douglas, Angela E

    2015-09-01

    Gut microorganisms are essential for the nutritional health of many animals, but the underlying mechanisms are poorly understood. This study investigated how lipid accumulation by adult Drosophila melanogaster is reduced in flies associated with the bacterium Acetobacter tropicalis which displays oral-faecal cycling between the gut and food. We demonstrate that the lower lipid content of A. tropicalis-colonized flies relative to bacteria-free flies is linked with a parallel bacterial-mediated reduction in food glucose content; and can be accounted for quantitatively by the amount of glucose acquired by the flies, as determined from the feeding rate and assimilation efficiency of bacteria-free and A. tropicalis-colonized flies. We recommend that nutritional studies on Drosophila include empirical quantification of food nutrient content, to account for likely microbial-mediated effects on diet composition. More broadly, this study demonstrates that selective consumption of dietary constituents by microorganisms can alter the nutritional balance of food and, thereby, influence the nutritional status of the animal host. PMID:26382071

  20. Determination of the primary structure of two lipid transfer proteins from apricot (Prunus armeniaca).

    PubMed

    Conti, A; Fortunato, D; Ortolani, C; Giuffrida, M G; Pravettoni, V; Napolitano, L; Farioli, L; Perono Garoffo, L; Trambaioli, C; Pastorello, E A

    2001-05-25

    It has been recently demonstrated that the major allergen of apricot is a protein of molecular mass (Mr) 9000 belonging to the family of Lipid Transfer Protein. The aim of this study was the determination of the primary structure of apricot LTP by micro-sequencing and mass spectrometric analyses. Apricot LTP is a 91 amino acids protein like peach and almond LTPs with a sequence identity of 91% and 94%, respectively. Like for the peach LTP, out of the 25 amino acids forming the inner surface of the tunnel-like hydrophobic cavity in maize ns-LTP, 16 are identical and 7 similar in the apricot LTP, supporting the hypothesis of a similar function.

  1. Determination of orientational order parameters from 2H NMR spectra of magnetically partially oriented lipid bilayers.

    PubMed Central

    Schäfer, H; Mädler, B; Sternin, E

    1998-01-01

    The partial orientation of multilamellar vesicles (MLVs) in high magnetic fields is known to affect the shape of 2H NMR spectra. There are numerical methods for extracting either the orientational order parameters of lipid molecules for a random distribution of domain orientations in the sample, or the distribution of orientations for a known set of spectral anisotropies. A first attempt at determining the orientational order parameters in the presence of an unknown nonrandom distribution of orientations is presented. The numerical method is based on the Tikhonov regularization algorithm. It is tested using simulated partially oriented spectra. An experimental spectrum of a phospholipid-ether mixture in water is analyzed as an example. The experimental spectrum is consistent with an ellipsoidal shape of MLVs with a ratio of semiaxes of approximately 3.4. PMID:9533713

  2. Determination of lipid A and endotoxin in serum by mass spectroscopy

    PubMed Central

    Maitra, Shyamal K.; Schotz, Michael C.; Yoshikawa, Thomas T.; Guze, Lucien B.

    1978-01-01

    A quantitative technique for determining lipid A content of endotoxin added to serum by combined gas chromatography-mass spectrometry is described. This technique uses detection of the β-hydroxymyristic acid content of Salmonella minnesota R595 lipopolysaccharide by selected ion monitoring at atomic mass unit of 315.4 The fatty acids produced on hydrolysis of serum containing lipopolysaccharide were extracted and the methyl esters were made. Silica gel chromatography was used to separate methyl esters of hydroxy fatty acids from other fatty acid methyl esters. Trimethylsilyl ether derivatives of the hydroxy fatty acid methyl ester fraction were quantitated by this technique. As little as 200 fmol of β-hydroxymyristic acid could be detected. PMID:16592555

  3. Determination of the topography of cytochrome b5 in lipid vesicles by fluorescence quenching.

    PubMed

    Markello, T; Zlotnick, A; Everett, J; Tennyson, J; Holloway, P W

    1985-06-01

    Cytochrome b5, a protein isolated from the endoplasmic reticulum by detergent extraction, interacts spontaneously with small unilamellar phosphatidylcholine vesicles. When the vesicles are made from 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), the tryptophan fluorescence of the cytochrome is enhanced, and when they are made from 1-palmitoyl-2-(dibromostearoyl) phosphatidylcholine (BRPC), the fluorescence is quenched. A series of BRPC were synthesized with bromine atoms at the 6,7, 9,10, 11,12 or 15,16 positions. The vesicles synthesized from each of these lipids were similar in size to those made from POPC. The relative fluorescence intensities of the cytochrome b5 in POPC and 6,7-, 9,10-, 11,12- and 15,16- BRPC were 100, 19.4, 29.4, 37.1, and 54.0, respectively. These data suggest that the exposed tryptophan(s) is (are) at a depth of 0.7 nm below the surface of the vesicle. Bromine is a collisional quencher; hence, these data may indicate the relative position of the lipid annulus around the protein rather than the depth of the protein below the average vesicle surface. Cytochrome b5 contains three potentially fluorescent tryptophans, and determinations of fluorescent quantum yield indicate all three potentially fluorescent tryptophans, and determinations of fluorescent quantum yield indicate all three are fluorescent with an average quantum yield, when in POPC vesicles, of 0.21. Fluorescence lifetime measurements by the demodulation technique indicated heterogeneity of fluorescence lifetimes in all vesicles. The lifetimes in the BRPC vesicles ranged from 2.0 to 2.4 ns compared to a value of 3.3 ns in POPC.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. Determination of Components in Beverages by Thin-Layer Chromatography.

    ERIC Educational Resources Information Center

    Ma, Yinfa; Yeung, Edward S.

    1990-01-01

    Described is a simple and interesting chromatography experiment using three different fluorescence detection principles for the determination of caffeine, saccharin and sodium benzoate in beverages. Experimental procedures and an analysis and discussion of the results are included. (CW)

  5. Steady-state electrochemical determination of lipidic nanotube diameter utilizing an artificial cell model.

    PubMed

    Adams, Kelly L; Engelbrektsson, Johan; Voinova, Marina; Zhang, Bo; Eves, Daniel J; Karlsson, Roger; Heien, Michael L; Cans, Ann-Sofie; Ewing, Andrew G

    2010-02-01

    By exploiting the capabilities of steady-state electrochemical measurements, we have measured the inner diameter of a lipid nanotube using Fick's first law of diffusion in conjunction with an imposed linear concentration gradient of electroactive molecules over the length of the nanotube. Fick's law has been used in this way to provide a direct relationship between the nanotube diameter and the measurable experimental parameters Deltai (change in current) and nanotube length. Catechol was used to determine the Deltai attributed to its flux out of the nanotube. Comparing the nanotube diameter as a function of nanotube length revealed that membrane elastic energy was playing an important role in determining the size of the nanotube and was different when the tube was connected to either end of two vesicles or to a vesicle on one end and a pipet tip on the other. We assume that repulsive interaction between neck regions can be used to explain the trends observed. This theoretical approach based on elastic energy considerations provides a qualitative description consistent with experimental data.

  6. Optimization of a Nile Red method for rapid lipid determination in autotrophic, marine microalgae is species dependent.

    PubMed

    Balduyck, Lieselot; Veryser, Cedrick; Goiris, Koen; Bruneel, Charlotte; Muylaert, Koenraad; Foubert, Imogen

    2015-11-01

    Several studies have been conducted to develop rapid methods for quantification of lipid content in microalgae, as an alternative for time consuming gravimetric methods. Different studies showed that lipid staining with Nile Red in whole cell suspensions and subsequently quantification by the use of a spectrofluorometric device is a promising method, but a profound optimization and validation is rare. It has already been proven that the correlation curve for quantification is species dependent, but it has not yet been investigated whether this is also the case for the optimization of the Nile Red assay protocol. Therefore, two autotrophic, marine microalgae, Nannochloropsis oculata and T-Isochrysis lutea, strongly differing in e.g. cell wall structure, were selected in this study to investigate whether optimization of the Nile Red assay is species dependent. Besides this, it was checked for one of these species, Nannochloropsis, whether the lipid content, determined by the Nile Red assay, could indeed be correlated with the neutral and/or total lipid content determined by gravimetric methods. It was found that optimization of the Nile Red assay was strongly species dependent. Consequently, optimization has to be done for each species before using the assay. For Nannochloropsis, a good correlation was found between total and neutral lipid content obtained by the Nile Red assay and by gravimetric methods. PMID:26388510

  7. Optimization of a Nile Red method for rapid lipid determination in autotrophic, marine microalgae is species dependent.

    PubMed

    Balduyck, Lieselot; Veryser, Cedrick; Goiris, Koen; Bruneel, Charlotte; Muylaert, Koenraad; Foubert, Imogen

    2015-11-01

    Several studies have been conducted to develop rapid methods for quantification of lipid content in microalgae, as an alternative for time consuming gravimetric methods. Different studies showed that lipid staining with Nile Red in whole cell suspensions and subsequently quantification by the use of a spectrofluorometric device is a promising method, but a profound optimization and validation is rare. It has already been proven that the correlation curve for quantification is species dependent, but it has not yet been investigated whether this is also the case for the optimization of the Nile Red assay protocol. Therefore, two autotrophic, marine microalgae, Nannochloropsis oculata and T-Isochrysis lutea, strongly differing in e.g. cell wall structure, were selected in this study to investigate whether optimization of the Nile Red assay is species dependent. Besides this, it was checked for one of these species, Nannochloropsis, whether the lipid content, determined by the Nile Red assay, could indeed be correlated with the neutral and/or total lipid content determined by gravimetric methods. It was found that optimization of the Nile Red assay was strongly species dependent. Consequently, optimization has to be done for each species before using the assay. For Nannochloropsis, a good correlation was found between total and neutral lipid content obtained by the Nile Red assay and by gravimetric methods.

  8. Determination of elastic properties of surface layers and coatings by resonant ultrasound spectroscopy

    NASA Astrophysics Data System (ADS)

    Růžek, M.; Sedlák, P.; Seiner, H.; Landa, M.

    2011-01-01

    This paper deals with determination of in-plane elastic constants of thin layers deposited on substrates. Modified resonant ultrasound spectroscopy is used to measure resonant spectra before and after layer deposition . These two spectra are compared and changes in the position of the resonant peaks are associated with layer properties. It is shown that for thin layers either the elastic moduli or the surface mass density can be determined, providing the complementary information (the surface mass density for the determination of the moduli, the elastic moduli for the determination of the surface mass density) is known. As an experimental demonstration of this approach, the elastic moduli of diamond-like-carbon film deposited on a silicon substrate and the surface mass density of a thin spray paint on a silicon substrate are determined.

  9. Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods

    EPA Science Inventory

    Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [E. Van Handel, J. Am. Mosq. Control Assoc. 1(1985) 302] and recently validated for small samples by Inouye and Lotufo ...

  10. Development of novel fluorescent probe 3-perylene diphenylphosphine for determination of lipid hydroperoxide with fluorescent image analysis

    SciTech Connect

    Chotimarkorn, Chatchawan; Nagasaka, Reiko; Ushio, Hideki . E-mail: hushio@s.kaiyodai.ac.jp; Ohshima, Toshiaki; Matsunaga, Shigeki

    2005-12-16

    A novel fluorescent probe 3-perylene diphenylphosphine (3-PeDPP) was synthesized for the direct analysis of lipid hydroperoxides. The structure of 3-PeDPP was identified by the spectroscopic data, FAB-MS, {sup 1}H NMR, and {sup 13}C NMR. The reactivities of 3-PeDPP with lipid hydroperoxides were investigated in chloroform/MeOH homogeneous solutions and PC liposome model systems oxidized by either 2,2'-azobis(2-amidinopropane)dihydrochloride and photosensitized oxidation. The fluorescence intensity derived from 3-perylene diphenylphosphineoxide (3-PeDPPO) increased proportionally with amount of hydroperoxides produced in homogeneous solutions and liposome model systems. 3-PeDPP was easily incorporated into mouse myeloma SP2 cells and thin tissue section for dynamic membrane lipid peroxidation studies. Linear correlations between fluorescence intensity and amount of hydroperoxides in the cell membrane and tissue sections were obtained. The fluorescence intensity from 2-dimensional image analysis was also well correlated with lipid hydroperoxide level in these models. Thus, the novel probe 3-PeDPP is useful for the direct determination of lipid hydroperoxides in biological materials.

  11. Membrane Protein Structure Determination Using Crystallography and Lipidic Mesophases - Recent Advances and Successes

    PubMed Central

    Caffrey, Martin; Li, Dianfan; Dukkipati, Abhiram

    2012-01-01

    The crystal structure of the β2-adrenergic receptor in complex with an agonist and its cognate G protein has just recently been solved. It is now possible to explore in molecular detail the means by which this paradigmatic transmembrane receptor binds agonist, communicates the impulse or signalling event across the membrane and sets in motion a series of G protein-directed intracellular responses. The structure was determined using crystals of the ternary complex grown in a rationally designed lipidic mesophase by the so-called in meso method. The method is proving to be particularly useful in the G protein-coupled receptor field where the structures of thirteen distinct receptor types have been solved in the past five years. In addition to receptors, the method has proven useful with a wide variety of integral membrane protein classes that include bacterial and eukaryotic rhodopsins, a light harvesting complex II (LHII), photosynthetic reaction centers, cytochrome oxidases, β-barrels, an exchanger, and an integral membrane peptide. This attests to the versatility and range of the method and supports the view that the in meso method should be included in the arsenal of the serious membrane structural biologist. For this to happen however, the reluctance in adopting it attributable, in part, to the anticipated difficulties associated with handling the sticky, viscous cubic mesophase in which crystals grow must be overcome. Harvesting and collecting diffraction data with the mesophase-grown crystals is also viewed with some trepidation. It is acknowledged that there are challenges associated with the method. Over the years, we have endeavored to establish how the method works at a molecular level and to make it user-friendly. To these ends, tools for handling the mesophase in the pico- to nano-liter volume range have been developed for highly efficient crystallization screening in manual and robotic modes. Methods have been implemented for evaluating the functional

  12. Determination of lipid peroxidation and cytotoxicity in calcium, magnesium, titanium and hectorite (SHCa-1) suspensions.

    PubMed

    Kibanova, Daria; Nieto-Camacho, Antonio; Ramírez-Apan, Teresa; Cervini-Silva, Javiera

    2011-01-01

    This paper reports data on the relative ability of CaO, CaCl2, MgO, MgCl2, TiO2, and hectorite (SHCa-1) to induce oxidative stress (as determined by lipid peroxidation, LP) in biological matrices. The effectiveness of structural (oxide form) versus soluble Ca and Mg to induce LP is compared. An assessment on cytotoxicity as affected by soluble and structural Ca, Mg, TiO2 and SHCa-1 is also addressed. LP was screened and monitored using the Thiobarbituric Acid Reactive Substances (TBARS). The extent of TBARS production was found to vary with the type and initial concentration of the soluble or structural cation, Ca or Mg respectively. Obtained results showed higher magnitude values for the latter set of experiments. In the presence of TiO2 no significant TBARS production was detected pointing out a negligible effect of TiO2 on LP. At solid concentrations ca. 100 ppm, CaO appears to be more effective than SHCa-1 to induce LP. By contrast at ca. 25 ppm, MgO appears to be more effective than the clay mineral. The SHCa-1 LP-inducing activity has been proven to closely relate to structural Ca. The prevalence of mechanisms that may induce LP but not cytotoxicity (as determined by cell growth inhibition) was also addressed. Results on cell growth inhibition as affected by soluble and structural Ca, Mg, TiO2 and hectorite provide evidence to support that structural Ca or Mg brings about significantly higher variations than soluble Ca.

  13. Comparison of Methods for Determining Boundary Layer Edge Conditions for Transition Correlations

    NASA Technical Reports Server (NTRS)

    Liechty, Derek S.; Berry, Scott A.; Hollis, Brian R.; Horvath, Thomas J.

    2003-01-01

    Data previously obtained for the X-33 in the NASA Langley Research Center 20-Inch Mach 6 Air Tunnel have been reanalyzed to compare methods for determining boundary layer edge conditions for use in transition correlations. The experimental results were previously obtained utilizing the phosphor thermography technique to monitor the status of the boundary layer downstream of discrete roughness elements via global heat transfer images of the X-33 windward surface. A boundary layer transition correlation was previously developed for this data set using boundary layer edge conditions calculated using an inviscid/integral boundary layer approach. An algorithm was written in the present study to extract boundary layer edge quantities from higher fidelity viscous computational fluid dynamic solutions to develop transition correlations that account for viscous effects on vehicles of arbitrary complexity. The boundary layer transition correlation developed for the X-33 from the viscous solutions are compared to the previous boundary layer transition correlations. It is shown that the boundary layer edge conditions calculated using an inviscid/integral boundary layer approach are significantly different than those extracted from viscous computational fluid dynamic solutions. The present results demonstrate the differences obtained in correlating transition data using different computational methods.

  14. Microfluidic Synthesis of Hybrid Nanoparticles with Controlled Lipid Layers: Understanding Flexibility-Regulated Cell-Nanoparticle Interaction.

    PubMed

    Zhang, Lu; Feng, Qiang; Wang, Jiuling; Zhang, Shuai; Ding, Baoquan; Wei, Yujie; Dong, Mingdong; Ryu, Ji-Young; Yoon, Tae-Young; Shi, Xinghua; Sun, Jiashu; Jiang, Xingyu

    2015-10-27

    The functionalized lipid shell of hybrid nanoparticles plays an important role for improving their biocompatibility and in vivo stability. Yet few efforts have been made to critically examine the shell structure of nanoparticles and its effect on cell-particle interaction. Here we develop a microfluidic chip allowing for the synthesis of structurally well-defined lipid-polymer nanoparticles of the same sizes, but covered with either lipid-monolayer-shell (MPs, monolayer nanoparticles) or lipid-bilayer-shell (BPs, bilayer nanoparticles). Atomic force microscope and atomistic simulations reveal that MPs have a lower flexibility than BPs, resulting in a more efficient cellular uptake and thus anticancer effect than BPs do. This flexibility-regulated cell-particle interaction may have important implications for designing drug nanocarriers.

  15. Determination of general relations for the behavior of turbulent boundary layers

    NASA Technical Reports Server (NTRS)

    Von Doenhoff, Albert E; Tetervin, Neal

    1943-01-01

    An analysis has been made of a considerable amount of data for turbulent boundary layers along wings and bodies of various shapes in order to determine the fundamental variables that control the development of turbulent boundary layers. It was found that the type of velocity distribution in the boundary layer could be expressed in terms of a single parameter. This parameter was chosen as the ratio of the displacement thickness to the momentum thickness of the boundary layer. The variables that control the development of the turbulent boundary layer apparently are: (1) the ratio of the nondimensional pressure gradient, expressed in terms of the local dynamic pressure outside the boundary layer and boundary-layer thickness, to the local skin-friction coefficient and (2) the shape of the boundary layer. An empirical equation has been developed in terms of these variables that, when used with the momentum equation and the skin-friction relation, makes it possible to trace the development of the turbulent boundary layer to the separation point.

  16. Examining the role of membrane lipid composition in determining the ethanol tolerance of Saccharomyces cerevisiae.

    PubMed

    Henderson, Clark M; Block, David E

    2014-05-01

    Yeast (Saccharomyces cerevisiae) has an innate ability to withstand high levels of ethanol that would prove lethal to or severely impair the physiology of other organisms. Significant efforts have been undertaken to elucidate the biochemical and biophysical mechanisms of how ethanol interacts with lipid bilayers and cellular membranes. This research has implicated the yeast cellular membrane as the primary target of the toxic effects of ethanol. Analysis of model membrane systems exposed to ethanol has demonstrated ethanol's perturbing effect on lipid bilayers, and altering the lipid composition of these model bilayers can mitigate the effect of ethanol. In addition, cell membrane composition has been correlated with the ethanol tolerance of yeast cells. However, the physical phenomena behind this correlation are likely to be complex. Previous work based on often divergent experimental conditions and time-consuming low-resolution methodologies that limit large-scale analysis of yeast fermentations has fallen short of revealing shared mechanisms of alcohol tolerance in Saccharomyces cerevisiae. Lipidomics, a modern mass spectrometry-based approach to analyze the complex physiological regulation of lipid composition in yeast and other organisms, has helped to uncover potential mechanisms for alcohol tolerance in yeast. Recent experimental work utilizing lipidomics methodologies has provided a more detailed molecular picture of the relationship between lipid composition and ethanol tolerance. While it has become clear that the yeast cell membrane composition affects its ability to tolerate ethanol, the molecular mechanisms of yeast alcohol tolerance remain to be elucidated.

  17. Determination of the transition point in a boundary layer by the measurement of pressure fluctuation

    NASA Technical Reports Server (NTRS)

    Strach, V.

    1981-01-01

    A method is developed for determining the laminar-to-turbulent transition point in a boundary layer. The method involves the measurement of pressure on the surface of the body and the computation of pressure fluctuations. This method of determining the transition point is suitable when visualization methods cannot be used.

  18. Direct determination of the thickness of stratospheric layers from single-channel satellite radiance measurements.

    NASA Technical Reports Server (NTRS)

    Quiroz, R. S.; Gelman, M. E.

    1972-01-01

    The direct use of measured radiances for determining the thickness of stratospheric layers is investigated. Layers based at 100-10 mb, with upper boundaries at 10-0.5 mb, are investigated using a carefully selected family of stratospheric temperature profiles and computed radiances. On the basis of physical reasoning, a high correlation of thickness with radiance is anticipated for deep layers, such as the 100- to 2-mb layer (from about 15 to 43 km), that emit a substantial part of the infrared energy reaching a satellite radiometer in a particular channel. Empirical regression curves relating thickness and radiance are developed and are compared with blackbody curves obtained by substituting the blackbody temperature in the hydrostatic equation. Maximum thickness-radiance correlation is found, for each infrared channel, for the layer having the best agreement of empirical and blackbody curves.

  19. Using fluorescence-activated flow cytometry to determine reactive oxygen species formation and membrane lipid peroxidation in viable boar spermatozoa.

    PubMed

    Guthrie, H David; Welch, Glenn R

    2010-01-01

    Fluorescence-activated flow cytometry analyses were developed for determination of reactive oxygen species (ROS) formation and membrane lipid peroxidation in live spermatozoa loaded with, respectively, hydroethidine (HE) or the lipophilic probe 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid, C(11)BODIPY(581/591) (BODIPY). ROS was detected by red fluorescence emission from oxidization of HE and membrane lipid peroxidation was detected by green fluorescence emission from oxidation of BODIPY in individual live sperm. Of the reactive oxygen species generators tested, BODIPY oxidation was specific for FeSo4/ascorbate (FeAc), because menadione and H(2)O(2) had little or no effect. The oxidization of hydroethidine to ethidium was specific for menadione and H(2)O(2); FeAc had no effect. The incidence of basal or spontaneous ROS formation and membrane lipid peroxidation were low in boar sperm (<1% of live sperm) in fresh semen or after low temperature storage; however the sperm were quite susceptible to treatment-induced ROS formation and membrane lipid peroxidation. PMID:20072917

  20. Formation of a suspended lipid membrane on a microcavity covered by a thin SiO2 layer with a nanohole array

    NASA Astrophysics Data System (ADS)

    Tanaka, Aya; Kashimura, Yoshiaki; Kuramochi, Eiichi; Sumitomo, Koji

    2014-01-01

    To provide a platform for a nanobiodevice, we fabricated microcavities on a Si/SiO2 substrate covered by a thin SiO2 layer with nanohole arrays that we call a pepper shaker substrate. Fluorescence and atomic force microscopy images showed that the structure of the pepper shaker substrate improved both the probability of membrane sealing over the microcavities by rupturing giant unilameller vesicles and the lifetime of the lipid membrane suspended over the microcavities. The success of this study reveals the potential for fabricating an artificial cell array as a tool for the functional and high throughput analysis of membrane proteins.

  1. Comparison of macro-gravimetric and micro-colorimetric lipid determination methods.

    PubMed

    Inouye, Laura S; Lotufo, Guiherme R

    2006-10-15

    In order to validate a method for lipid analysis of small tissue samples, the standard macro-gravimetric method of Bligh-Dyer (1959) [E.G. Bligh, W.J. Dyer, Can. J. Biochem. Physiol. 37 (1959) 911] and a modification of the micro-colorimetric assay developed by Van Handel (1985) [E. Van Handel, J. Am. Mosq. Control Assoc. 1 (1985) 302] were compared. No significant differences were observed for wet tissues of two species of fish. However, limited analysis of wet tissue of the amphipod, Leptocheirusplumulosus, indicated that the Bligh-Dyer gravimetric method generated higher lipid values, most likely due to the inclusion of non-lipid materials. Additionally, significant differences between the methods were observed with dry tissues, with the micro-colorimetric method consistently reporting calculated lipid values greater than as reported by the gravimetric method. This was most likely due to poor extraction of dry tissue in the standard Bligh-Dyer method, as no significant differences were found when analyzing a single composite extract. The data presented supports the conclusion that the micro-colorimetric method described in this paper is accurate, rapid, and minimizes time and solvent use.

  2. Postprandial lipid responses to standard carbohydrate challenges used to determine glycemic index values

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Prior studies assessing metabolic effects of different types of carbohydrate have focused on their glycemic response. Not considered has been the response of postprandial cardiometabolic risk indicators. This study assessed the postprandial lipid responses to two forms of carbohydrates used as ref...

  3. Determination of the Mixing Layer Height Over two Sites, Using Pilot Balloons During the MILAGRO Campaign

    NASA Astrophysics Data System (ADS)

    Wohrnschimmel, H.; Alonso, A. L.; Ángeles, F.; Sosa, G.; Varela, J.; Cárdenas, B.

    2007-12-01

    Among the mechanisms that affect air quality there is a variety of meteorological processes. An important process in this context are the changes in the mixing layer height during a day and over the year. The mixing layer height is the portion of the atmosphere close to the surface layer where air pollutants get diluted, without leaving this layer. Therefore, it is important to describe the variations in the height of the mixing layer, i.e. the vertical dilution of air pollution, since this is a process mitigating naturally the impact of emissions. There exist different methods to obtain information on the mixing layer height, among them radio soundings, the application of vertical wind profilers, and launching pilot balloons. In this study, pilot balloons have been used simultaneously over two sites of the Mexico City Metropolitan Area during the MILAGRO campaign in March 2006. The objective was to determine the vertical wind profiles and derive information on the mixing layer height. Daily, four pilot balloons were launched, at 9:00, 12:00, 15:00, and 18:00 hours, over Tenango del Aire (a rural area in the Southeast of Mexico City), and over Ciudad Universitaria, in the Southern metropolitan area. At some occasions, night time measurements have been carried out at 21:00 and 24:00. A variability of the diurnal evolution of the mixing layer was observed along March, which could be related to surface temperature. The diurnal evolution showed a sudden growth of the mixing layer between 9:00 and 12:00 hours. Data intercomparisons were carried out for pilot balloons versus radio soundings during a few days at a third site, Tula, in the North of Mexico City. Both intercomparisons showed that pilot balloons are an effective method to obtain information about the development of the mixing layer.

  4. Archaebacterial activity in the Orca Basin determined by the isolation of characteristic isopranyl ether-linked lipids

    NASA Astrophysics Data System (ADS)

    Dickins, Holli D.; Van Vleet, Edward S.

    1992-04-01

    Phytanyl glycerol ether lipids characteristic of archaebacterial inputs have been quantified in 30 water samples taken in the Orca Basin, an anoxic hypersaline basin located in the northwestern Gulf of Mexico. Because of the Basin's anoxic hypersaline character, it seems likely that archaebacteria may play a significant role in the microbial ecology of the brine. Physical data, including temperature, salinity, per cent transmission, oxygen and nutrient concentrations, also were collected from six depths at five sampling sites in the Basin. Four of the five sites were characterized by a 200 m thick, anoxic brine (salinity ≈ 250 ppt) at an approximate water depth of 2240 m. A stepwise increase in salinity was associated with the brine-seawater interface, increasing from 38 to 150 ppt within the upper portion of the 10 m interface and to >250 ppt within the brine. Three distinct layers of particulate material were observed within the 10 m interface. Corresponding with the salinity gradient was a decrease in dissolved oxygen from 5.0 ml l -1 at 2040 m to 0 ml l -1 within the brine. Ammonia and phosphate concentrations increased from 0 and 2.5 μM above the brine to 519 and 63.5 μM within the brine. At the same time, nitrate concentrations decreased from 22 μM above the brine to negligible within the brine. Depletion of oxygen, with concomitant increases in ammonia and phosphate, decreased nitrate, and the production of methane suggest microbially mediated processes may be occurring at the brine-seawater interface. Highest concentrations of phytanyl ether lipids were observed within the interface, ranging from 29.7 to 84.1 ng l -1. Concentrations were negligible below the interface. Elevated phytanyl ether lipid concentrations in conjunction with microbial activity studies carried out by other investigators suggest that archaebacterial activity is occurring within the brine particulate layers. A decline in ether lipid concentration and microbial activity below this

  5. Determination of lipid oxidation products in vegetable oils and marine omega-3 supplements

    PubMed Central

    Halvorsen, Bente Lise; Blomhoff, Rune

    2011-01-01

    Background There is convincing evidence that replacing dietary saturated fats with polyunsaturated fats (PUFA) decreases risk of cardiovascular diseases. Therefore, PUFA rich foods such as vegetable oils, fatty fish, and marine omega-3 supplements are recommended. However, PUFA are easily oxidizable and there is concern about possible negative health effects from intake of oxidized lipids. Little is known about the degree of lipid oxidation in such products. Objective To assess the content of lipid oxidation products in a large selection of vegetable oils and marine omega-3 supplements available in Norway. Both fresh and heated vegetable oils were studied. Design A large selection of commercially available vegetable oils and marine omega-3 supplements was purchased from grocery stores, pharmacies, and health food stores in Norway. The content of lipid oxidation products were measured as peroxide value and alkenal concentration. Twelve different vegetable oils were heated for a temperature (225°C) and time (25 minutes) resembling conditions typically used during cooking. Results The peroxide values were in the range 1.04–10.38 meq/kg for omega-3 supplements and in the range 0.60–5.33 meq/kg for fresh vegetable oils. The concentration range of alkenals was 158.23–932.19 nmol/mL for omega-3 supplements and 33.24–119.04 nmol/mL for vegetable oils. After heating, a 2.9–11.2 fold increase in alkenal concentration was observed for vegetable oils. Conclusions The contents of hydroperoxides and alkenals in omega-3 supplements are higher than in vegetable oils. After heating vegetable oils, a large increase in alkenal concentration was observed. PMID:21691461

  6. Gramicidin-based fluorescence assay; for determining small molecules potential for modifying lipid bilayer properties.

    PubMed

    Ingólfsson, Helgi I; Sanford, R Lea; Kapoor, Ruchi; Andersen, Olaf S

    2010-10-13

    Many drugs and other small molecules used to modulate biological function are amphiphiles that adsorb at the bilayer/solution interface and thereby alter lipid bilayer properties. This is important because membrane proteins are energetically coupled to their host bilayer by hydrophobic interactions. Changes in bilayer properties thus alter membrane protein function, which provides an indirect way for amphiphiles to modulate protein function and a possible mechanism for "off-target" drug effects. We have previously developed an electrophysiological assay for detecting changes in lipid bilayer properties using linear gramicidin channels as probes. Gramicidin channels are mini-proteins formed by the transbilayer dimerization of two non-conducting subunits. They are sensitive to changes in their membrane environment, which makes them powerful probes for monitoring changes in lipid bilayer properties as sensed by bilayer spanning proteins. We now demonstrate a fluorescence assay for detecting changes in bilayer properties using the same channels as probes. The assay is based on measuring the time-course of fluorescence quenching from fluorophore-loaded large unilamellar vesicles due to the entry of a quencher through the gramicidin channels. We use the fluorescence indicator/quencher pair 8-aminonaphthalene-1,3,6-trisulfonate (ANTS)/Tl(+) that has been successfully used in other fluorescence quenching assays. Tl(+) permeates the lipid bilayer slowly but passes readily through conducting gramicidin channels. The method is scalable and suitable for both mechanistic studies and high-throughput screening of small molecules for bilayer-perturbing, and potential "off-target", effects. We find that results using this method are in good agreement with previous electrophysiological results.

  7. Anionic Lipids: Determinants of Binding Cytotoxins from Snake Venom on the Surface of Cell Membranes

    PubMed Central

    Boldyrev, I.A.; Omelkov, A.V.; Utkin, Yu.N.; Efremov, R.G.

    2010-01-01

    The cytotoxic properties of cytotoxins (CTs) from snake venom are mediated by their interaction with the cell membrane. The hydrophobic pattern containing the tips of loops I–III and flanked by polar residues is known to be a membrane–binding motif of CTs. However, this is not enough to explain the difference in activity among various CTs which are similar in sequence and in 3D structure. The mechanism of further CT–membrane interaction leading to pore formation and cell death still remains unknown. Published experimental data on the specific interaction between CT and low molecular weight anionic components (sulphatide) of the bilayer point to the existence of corresponding ligand binding sites on the surface of toxin molecules. In this work we study the membrane–lytic properties of CT I, CT II (Naja oxiana), and Ct 4 (Naja kaouthia), which belong to different structural and functional types (P– and S–type) of CTs, by measuring the intensity of a fluorescent dye, calcein released from liposomes containing a phosphatidylserine (PS) lipid as an anionic component. Using molecular docking simulations, we find and characterize three sites in CT molecules that can potentially bind the PS polar head. Based on the data obtained, we suggest a hypothesis that CTs can specifically interact with one or more of the anionic lipids (in particular, with PS) contained in the membrane, thus facilitating the interaction between CTs and the lipid bilayer of a cell membrane. PMID:22649646

  8. Lipid and fatty acid analysis of the Plodia interpunctella granulosis virus (PiGV) envelope

    NASA Technical Reports Server (NTRS)

    Shastri-Bhalla, K.; Funk, C. J.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    Virus envelope was isolated from Plodia interpunctella granulosis virus, produced in early fourth-instar larvae. Both polar and neutral lipids were analyzed by two-dimensional thin-layer chromatography. Fatty acid composition of various individual neutral and polar lipids was determined by gas-liquid chromatography. The major components of envelope neutral lipid were diacylglycerols. Palmitic acid and stearic acid were the major saturated fatty acids in both polar and neutral lipids. Whereas palmitoleic acid was the major unsaturated fatty acids in neutral lipids, oleic acid was the major unsaturated fatty acid in the polar lipids.

  9. Determination of lipid ester ozonides and core aldehydes by high-performance liquid chromatography with on-line mass spectrometry.

    PubMed

    Ravandi, A; Kuksis, A; Myher, J J; Marai, L

    1995-11-01

    Unsaturated triacylglycerols (TG) and choline (PC) and ethanolamine (PE) phosphatides of known structure were subjected to ozonization and reduction with triphenylphosphine to yield the corresponding lipid ester core aldehydes. Mono- and di-C9 aldehyde palmitoylglycerols were prepared from oleoyldipalmitoyl and oleoyllinoleoylpalmitoyl glycerols, respectively, while egg yolk PC and PE provided the mono-C5 and mono-C9 aldehydes of palmitoyl-and stearoyl glycerophospholipids. The aldehydes were isolated in the free form and as the dinitrophenylhydrazone (DNPH) derivatives by thin-layer chromatography (TLC). The intermediate ozonides, free aldehydes and hydrazones were identified by reversed phase high performance liquid chromatography (HPLC) with on-line negative ion thermospray and normal phase HPLC with on-line positive ion electrospray mass spectrometry (LC-MS). The synthetic aldehydes were used as carriers during isolation from natural sources and as reference compounds in quantitative analyses.

  10. Determining the imaging plane of a retinal capillary layer in adaptive optical imaging

    NASA Astrophysics Data System (ADS)

    Yang, Le-Bao; Hu, Li-Fa; Li, Da-Yu; Cao, Zhao-Liang; Mu, Quan-Quan; Ma, Ji; Xuan, Li

    2016-09-01

    Even in the early stage, endocrine metabolism disease may lead to micro aneurysms in retinal capillaries whose diameters are less than 10 μm. However, the fundus cameras used in clinic diagnosis can only obtain images of vessels larger than 20 μm in diameter. The human retina is a thin and multiple layer tissue, and the layer of capillaries less than 10 μm in diameter only exists in the inner nuclear layer. The layer thickness of capillaries less than 10 μm in diameter is about 40 μm and the distance range to rod&cone cell surface is tens of micrometers, which varies from person to person. Therefore, determining reasonable capillary layer (CL) position in different human eyes is very difficult. In this paper, we propose a method to determine the position of retinal CL based on the rod&cone cell layer. The public positions of CL are recognized with 15 subjects from 40 to 59 years old, and the imaging planes of CL are calculated by the effective focal length of the human eye. High resolution retinal capillary imaging results obtained from 17 subjects with a liquid crystal adaptive optics system (LCAOS) validate our method. All of the subjects’ CLs have public positions from 127 μm to 147 μm from the rod&cone cell layer, which is influenced by the depth of focus. Project supported by the National Natural Science Foundation of China (Grant Nos. 11174274, 11174279, 61205021, 11204299, 61475152, and 61405194).

  11. Determining the imaging plane of a retinal capillary layer in adaptive optical imaging

    NASA Astrophysics Data System (ADS)

    Yang, Le-Bao; Hu, Li-Fa; Li, Da-Yu; Cao, Zhao-Liang; Mu, Quan-Quan; Ma, Ji; Xuan, Li

    2016-09-01

    Even in the early stage, endocrine metabolism disease may lead to micro aneurysms in retinal capillaries whose diameters are less than 10 μm. However, the fundus cameras used in clinic diagnosis can only obtain images of vessels larger than 20 μm in diameter. The human retina is a thin and multiple layer tissue, and the layer of capillaries less than 10 μm in diameter only exists in the inner nuclear layer. The layer thickness of capillaries less than 10 μm in diameter is about 40 μm and the distance range to rod&cone cell surface is tens of micrometers, which varies from person to person. Therefore, determining reasonable capillary layer (CL) position in different human eyes is very difficult. In this paper, we propose a method to determine the position of retinal CL based on the rod&cone cell layer. The public positions of CL are recognized with 15 subjects from 40 to 59 years old, and the imaging planes of CL are calculated by the effective focal length of the human eye. High resolution retinal capillary imaging results obtained from 17 subjects with a liquid crystal adaptive optics system (LCAOS) validate our method. All of the subjects’ CLs have public positions from 127 μm to 147 μm from the rod&cone cell layer, which is influenced by the depth of focus. Project supported by the National Natural Science Foundation of China (Grant Nos. 11174274, 11174279, 61205021, 11204299, 61475152, and 61405194).

  12. Role of Burkholderia cenocepacia afcE and afcF genes in determining lipid-metabolism-associated phenotypes.

    PubMed

    Subramoni, Sujatha; Agnoli, Kirsty; Eberl, Leo; Lewenza, Shawn; Sokol, Pamela A

    2013-03-01

    Burkholderia cenocepacia is an opportunistic pathogen that primarily infects cystic fibrosis patients. Previously we have reported that mutations in shvR, a LysR-type transcriptional regulator, and ShvR-regulated genes BCAS0208 and BCAS0201 (designated afcE and afcF, respectively) affect colony morphotype, biofilm and pellicle formation and virulence in B. cenocepacia. In this study we investigated the role of afcE and afcF in influencing lipid-metabolism-associated phenotypes. As previously reported for K56-2ΔshvR, the Δ2afcE and afcF : : lux mutants had no antifungal activity against Fusarium and Rhizoctonia solani, suggesting that these genes are involved in synthesis of a membrane-associated antifungal lipopeptide. Strains Δ2afcE and afcF : : lux had reduced swarming motility and altered cell membrane morphology, both of which were restored to wild-type levels upon providing these genes in trans. Both K56-2ΔshvR and Δ2afcE showed increased uptake of the hydrophobic fluorescent probe N-phenylnaphthylamine (NPN), indicating altered outer membrane properties. Total lipid profiles determined by TLC revealed distinct differences in cellular lipid compositions of K56-2ΔshvR, Δ2afcE and afcF : : lux compared with K56-2. Taken together, these results indicate that afcE and afcF are involved in metabolic pathway(s) influencing lipid profiles and affect both cell surface and antifungal properties of B. cenocepacia.

  13. Effect of Dietary Marine Microalgae (Schizochytrium) Powder on Egg Production, Blood Lipid Profiles, Egg Quality, and Fatty Acid Composition of Egg Yolk in Layers

    PubMed Central

    Park, J. H.; Upadhaya, S. D.; Kim, I. H.

    2015-01-01

    Two hundred and sixteen Institut de Sélection Animale (ISA) brown layers (40 wks of age) were studied for 6 wks to examine the effect of microalgae powder (MAP) on egg production, egg quality, blood lipid profile, and fatty acid concentration of egg yolk. Dietary treatments were as follows: i) CON (basal diet), ii) 0.5% MAP (CON+0.5% Schizochytrium powder), and iii) 1.0% MAP (CON+1.0% Schizochytrium powder). From 44 to 46 wks, egg production was higher in 1.0% MAP treatment than in control treatment (linear, p = 0.034); however, there was no difference on the egg production from 40 to 43 wks (p>0.05). Serum triglyceride and total cholesterol were significantly reduced in the groups fed with MAP, compared to those in groups fed with control diets (Quadratic, p = 0.034 and p = 0.039, respectively). Inclusion of 0.5% MAP in the diet of layers improved egg yolk color, compared with hens fed with basal diet at 46 wks (quadratic, p = 0.044). Eggshell thickness was linearly increased in MAP-fed treatments at 46th wk (p<0.05). Concentration of yolk docosahexaenoic acid (DHA; C22:6n-3) was increased in treatment groups fed with MAP (linear, p<0.05). The n-6 fatty acids, n-6/n-3 fatty acid, and unsaturated fatty acid/saturated fatty acid were decreased in treatment groups fed with MAP (linear, p<0.05). These results suggest that MAP improved the egg production and egg quality, and may affect serum lipid metabolites in the layers. In addition, MAP increases yolk DHA levels, and deceases n-6/n-3 fatty acid ratio. PMID:25656210

  14. Lipid composition determines the effects of arbutin on the stability of membranes.

    PubMed Central

    Hincha, D K; Oliver, A E; Crowe, J H

    1999-01-01

    Arbutin (hydroquinone-beta-D-glucopyranoside) is an abundant solute in the leaves of many freezing- or desiccation-tolerant plants. Its physiological role in plants, however, is not known. Here we show that arbutin protects isolated spinach (Spinacia oleracea L.) thylakoid membranes from freeze-thaw damage. During freezing of liposomes, the presence of only 20 mM arbutin led to complete leakage of a soluble marker from egg PC (EPC) liposomes. When the nonbilayer-forming chloroplast lipid monogalactosyldiacylglycerol (MGDG) was included in the membranes, this leakage was prevented. Inclusion of more than 15% MGDG into the membranes led to a strong destabilization of liposomes during freezing. Under these conditions arbutin became a cryoprotectant, as only 5 mM arbutin reduced leakage from 75% to 20%. The nonbilayer lipid egg phosphatidylethanolamine (EPE) had an effect similar to that of MGDG, but was much less effective, even at concentrations up to 80% in EPC membranes. Arbutin-induced leakage during freezing was accompanied by massive bilayer fusion in EPC and EPC/EPE membranes. Twenty percent MGDG in EPC bilayers completely inhibited the fusogenic effect of arbutin. The membrane surface probes merocyanine 540 and 2-(6-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)hexanoyl-1-hexadecanoyl-sn-glycero-3-phosph ocholi ne (NBD-C(6)-HPC) revealed that arbutin reduced the ability of both probes to partition into the membranes. Steady-state anisotropy measurements with probes that localize at different positions in the membranes showed that headgroup mobility was increased in the presence of arbutin, whereas the mobility of the fatty acyl chains close to the glycerol backbone was reduced. This reduction, however, was not seen in membranes containing 20% MGDG. The effect of arbutin on lipid order was limited to the interfacial region of the membranes and was not evident in the hydrophobic core region. From these data we were able to derive a physical model of the perturbing

  15. Determination of phosphatidylglycerol in amniotic fluid by a simple one-dimensional thin-layer chromatography method.

    PubMed

    Tsao, F H; Zachman, R D

    1982-01-01

    Phosphatidylglycerol (PG) in amniotic fluid is the second important component of lung surfactant phospholipids and may be clinically useful in assessing fetal lung maturity in utero. Although methods for PG determination are available, there are shortcomings in clinical application. We developed an alternative reliable one-dimensional thin-layer chromatography(TLC) method for separating and quantitating PG in amniotic fluid. A mini-TLC plate (8 X 10 cm) was prepared from silica gel H containing 5% ammonium sulfate. The plate was first developed in tetrahydrofuron/dimethoxymethane/methanol/2N ammonium hydroxide (30.0:20.6:5.6:3.0 v/v) and then in chloroform/methanol (60.9, v/v) in the same direction. PG was clearly separated from other phospholipids and neutral lipids, even when large amounts of other phospholipids were present on the TLC plate. The density of the charred PG was directly proportional to the amount of PG up to 8 nmol. The content of PG in nmol in the specimen can be quantitated by comparing with a standard PG. Up to 10% of blood serum or 3% meconium showed no detectable PG, nor did these substances affect PG quantitation in amniotic fluid. This method is sensitive and accurate. It is also time-saving and economical. PMID:7053902

  16. Layers

    NASA Astrophysics Data System (ADS)

    Hong, K. J.; Jeong, T. S.; Youn, C. J.

    2014-09-01

    The temperature-dependent photoresponse characteristics of MnAl2S4 layers have been investigated, for the first time, by use of photocurrent (PC) spectroscopy. Three peaks were observed at all temperatures. The electronic origin of these peaks was associated with band-to-band transitions from the valence-band states Γ4( z), Γ5( x), and Γ5( y) to the conduction-band state Γ1( s). On the basis of the relationship between PC-peak energy and temperature, the optical band gap could be well expressed by the expression E g( T) = E g(0) - 2.80 × 10-4 T 2/(287 + T), where E g(0) was estimated to be 3.7920 eV, 3.7955 eV, and 3.8354 eV for the valence-band states Γ4( z), Γ5( x), and Γ5( y), respectively. Results from PC spectroscopy revealed the crystal-field and spin-orbit splitting were 3.5 meV and 39.9 meV. The gradual decrease of PC intensity with decreasing temperature can be explained on the basis of trapping centers associated with native defects in the MnAl2S4 layers. Plots of log J ph, the PC current density, against 1/ T, revealed a dominant trap level in the high-temperature region. By comparing PC and the Hall effect results, we confirmed that this trap level is a shallow donor 18.9 meV below the conduction band.

  17. Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein

    PubMed Central

    Abdul Halim, Mohd Farid; Karch, Kelly R.; Zhou, Yitian; Haft, Daniel H.; Garcia, Benjamin A.

    2015-01-01

    ABSTRACT For years, the S-layer glycoprotein (SLG), the sole component of many archaeal cell walls, was thought to be anchored to the cell surface by a C-terminal transmembrane segment. Recently, however, we demonstrated that the Haloferax volcanii SLG C terminus is removed by an archaeosortase (ArtA), a novel peptidase. SLG, which was previously shown to be lipid modified, contains a C-terminal tripartite structure, including a highly conserved proline-glycine-phenylalanine (PGF) motif. Here, we demonstrate that ArtA does not process an SLG variant where the PGF motif is replaced with a PFG motif (slgG796F,F797G). Furthermore, using radiolabeling, we show that SLG lipid modification requires the PGF motif and is ArtA dependent, lending confirmation to the use of a novel C-terminal lipid-mediated protein-anchoring mechanism by prokaryotes. Similar to the case for the ΔartA strain, the growth, cellular morphology, and cell wall of the slgG796F,F797G strain, in which modifications of additional H. volcanii ArtA substrates should not be altered, are adversely affected, demonstrating the importance of these posttranslational SLG modifications. Our data suggest that ArtA is either directly or indirectly involved in a novel proteolysis-coupled, covalent lipid-mediated anchoring mechanism. Given that archaeosortase homologs are encoded by a broad range of prokaryotes, it is likely that this anchoring mechanism is widely conserved. IMPORTANCE Prokaryotic proteins bound to cell surfaces through intercalation, covalent attachment, or protein-protein interactions play critical roles in essential cellular processes. Unfortunately, the molecular mechanisms that anchor proteins to archaeal cell surfaces remain poorly characterized. Here, using the archaeon H. volcanii as a model system, we report the first in vivo studies of a novel protein-anchoring pathway involving lipid modification of a peptidase-processed C terminus. Our findings not only yield important insights into

  18. Experimental determination and computational interpretation of biophysical properties of lipid bilayers enriched by cholesteryl hemisuccinate.

    PubMed

    Kulig, Waldemar; Jurkiewicz, Piotr; Olżyńska, Agnieszka; Tynkkynen, Joona; Javanainen, Matti; Manna, Moutusi; Rog, Tomasz; Hof, Martin; Vattulainen, Ilpo; Jungwirth, Pavel

    2015-02-01

    Cholesteryl hemisuccinate (CHS) is one of the cholesterol-mimicking detergents not observed in nature. It is, however, widely used in protein crystallography, in biochemical studies of proteins, and in pharmacology. Here, we performed an extensive experimental and theoretical study on the behavior of CHS in lipid membranes rich in unsaturated phospholipids. We found that the deprotonated form of CHS (that is the predominant form under physiological conditions) does not mimic cholesterol very well. The protonated form of CHS does better in this regard, but also its ability to mimic the physical effects of cholesterol on lipid membranes is limited. Overall, although ordering and condensing effects characteristic to cholesterol are present in systems containing any form of CHS, their strength is appreciably weaker compared to cholesterol. Based on the considerable amount of experimental and atomistic simulation data, we conclude that these differences originate from the fact that the ester group of CHS does not anchor it in an optimal position at the water-membrane interface. The implications of these findings for considerations of protein-cholesterol interactions are briefly discussed. PMID:25450348

  19. Experimental determination and computational interpretation of biophysical properties of lipid bilayers enriched by cholesteryl hemisuccinate.

    PubMed

    Kulig, Waldemar; Jurkiewicz, Piotr; Olżyńska, Agnieszka; Tynkkynen, Joona; Javanainen, Matti; Manna, Moutusi; Rog, Tomasz; Hof, Martin; Vattulainen, Ilpo; Jungwirth, Pavel

    2015-02-01

    Cholesteryl hemisuccinate (CHS) is one of the cholesterol-mimicking detergents not observed in nature. It is, however, widely used in protein crystallography, in biochemical studies of proteins, and in pharmacology. Here, we performed an extensive experimental and theoretical study on the behavior of CHS in lipid membranes rich in unsaturated phospholipids. We found that the deprotonated form of CHS (that is the predominant form under physiological conditions) does not mimic cholesterol very well. The protonated form of CHS does better in this regard, but also its ability to mimic the physical effects of cholesterol on lipid membranes is limited. Overall, although ordering and condensing effects characteristic to cholesterol are present in systems containing any form of CHS, their strength is appreciably weaker compared to cholesterol. Based on the considerable amount of experimental and atomistic simulation data, we conclude that these differences originate from the fact that the ester group of CHS does not anchor it in an optimal position at the water-membrane interface. The implications of these findings for considerations of protein-cholesterol interactions are briefly discussed.

  20. Characterization of the Lipids of Butyrivibrio fibrisolvens

    PubMed Central

    Kunsman, Joseph E.

    1970-01-01

    Butyrivibrio fibrisolvens strain D-1 was grown on a lipid-free chemically defined medium. The lipids were extracted with chloroform-methanol and separated into nonpolar and polar fractions by silicic acid column chromatography. Further separations were made by preparative thin-layer chromatography. The lipid fractions were identified by specific staining reactions and RF values, by phosphorus and nitrogen determinations, by chromatography of hydrolysis products, and by the use of infrared spectroscopy. The major nonpolar lipid was free fatty acid. Four major polar lipids were identified: phosphatidylethanolamine, phosphatidyl glycerol, lipoaminoacid, and glycolipid. The lipoaminoacid contained alanine, leucine, and isoleucine. The glycolipid contained galactose. The major fatty acids identified were C16:0 and C18:1. The significance of the presence of lipoaminoacid is discussed. PMID:4316361

  1. [The preparation procedure of tests for the gas chromatographic determination of fat acids without preliminary extraction of lipids].

    PubMed

    Aripovskiĭ, A V; Kolesnik, P O; Vezhdel, M I; Titov, V N

    2012-01-01

    The enhancement of the procedure of quantitative gas chromatographic determination of fit acids in biologic liquids samples is proposed. Instead of the conventional Folch procedure of extraction of lipids with subsequent ablution, concentration and methylation of extracts the direct saponification and methylation of vacuum dried liquid samples (50-200 mkl) can be applied. To compare the effectiveness of the proposed and conventional procedures both of them had been applied to evaluate how converge the results of determination of composition of fat acids in whole blood, blood plasma, packed red blood cells, homogenates of hepatic and muscular tissues. The proposed procedure is applied to determine the characteristics of fat acids composition inpatients with ischemic heart disease.

  2. Lipids of Sarcina lutea III. Composition of the Complex Lipids

    PubMed Central

    Huston, Charles K.; Albro, Phillip W.; Grindey, Gerald B.

    1965-01-01

    Huston, Charles K. (Fort Detrick, Frederick, Md.), Phillip W. Albro, and Gerald B. Grindey. Lipids of Sarcina lutea. III. Composition of the complex lipids. J. Bacteriol. 89:768–775. 1965.—The complex lipids from a strain of Sarcina lutea were isolated and separated into fractions on diethylaminoethyl cellulose acetate and silicic acid columns. These fractions were monitored in several thin-layer chromatography systems. The various lipid types were characterized by their behavior in thin-layer systems and by an analysis of their hydrolysis products. The fatty acid composition of the column fractions was determined by gas-liquid chromatography. A number of components (13) were separated by thin-layer chromatography and characterized. The major components were polyglycerol phosphatide (17.0%), lipoamino acids (15.1%), phosphatidyl glycerol (13.8%), and an incompletely characterized substance (15.0%). Minor constituents included phosphatidyl inositol (5.5%), phosphatidic acid (4.2%), phosphatidyl serine (2.0%), and phosphatidyl choline (1.0%). No phosphatidyl ethanolamine was observed. PMID:14273659

  3. Oceanic upper mixed layer depth determination by the use of satellite data

    NASA Technical Reports Server (NTRS)

    Yan, Xiao-Hai; Schubel, J. R.; Pritchard, D. W.

    1990-01-01

    A method has been developed to determine the oceanic daily mean mixed layer depth from satellite observations and a mixed layer thermal inertia (MLTI) model. The algorithms were developed to use remotely-sensed values of sea surface temperature, albedo, and surface wind speeds to calculate the thermal inertia and to predict changes in subsurface diurnal mixed layer depth. The MLTI model, based on a mixed layer model of the upper ocean, has been used to simulate the diurnal mixing process and thermal inertia distribution in the Sargasso Sea around 34 deg N, 70 deg W. Sea surface temperature and albedo have been obtained from the NOAA7-AVHRR images. Surface wind speeds have been derived from the Scanning Multichannel Microwave Radiometer (SMMR) aboard Nimbus 7. Image processing was performed for images gathered between June and July 1982. The daily mean mixed layer depths predicted by the MLTI model agree well with data gathered at the LOTUS mooring located in the Sargasso Sea. This suggests that vertical mixing is the dominant physical process that controls the thermal inertia distribution in the midocean, far from major current systems, and that remote sensing is a promising tool to study such upper ocean processes.

  4. On determining characteristic length scales in pressure gradient turbulent boundary layers

    NASA Astrophysics Data System (ADS)

    Vinuesa, Ricardo; Örlü, Ramis; Schlatter, Philipp

    2016-04-01

    In the present work we analyze three methods used to determine the edge of pressure gradient turbulent boundary layers: two based on composite profiles, the one by Chauhan et al. (Fluid Dyn. Res. 41:021401, 2009) and the one by Nickels (J. Fluid Mech. 521:217–239, 2004), and the other one based on the condition of vanishing mean velocity gradient. Additionally, a new method is introduced based on the diagnostic plot concept by Alfredsson et al. (Phys. Fluids 23:041702, 2011). The boundary layer developing over the suction side of a NACA4412 wing profile, extracted from a direct numerical simulation at Rec = 400,000, is used as the test case. We find that all the methods produce robust results with mild or moderate pressure gradients, but stronger pressure gradients (with β larger than around 7) lead to inconsistent results in all the techniques except the diagnostic plot. This method also has the advantage of providing an objective way of defining the point where the mean streamwise velocity is 99% of the edge velocity, and shows consistent results in a wide range of pressure gradient conditions, as well as flow histories. Therefore, the technique based on the diagnostic plot is a robust method to determine the boundary layer thickness (equivalent to δ99) and edge velocity in pressure gradient turbulent boundary layers.

  5. On determining characteristic length scales in pressure gradient turbulent boundary layers

    NASA Astrophysics Data System (ADS)

    Vinuesa, Ricardo; Örlü, Ramis; Schlatter, Philipp

    2016-04-01

    In the present work we analyze three methods used to determine the edge of pressure gradient turbulent boundary layers: two based on composite profiles, the one by Chauhan et al. (Fluid Dyn. Res. 41:021401, 2009) and the one by Nickels (J. Fluid Mech. 521:217-239, 2004), and the other one based on the condition of vanishing mean velocity gradient. Additionally, a new method is introduced based on the diagnostic plot concept by Alfredsson et al. (Phys. Fluids 23:041702, 2011). The boundary layer developing over the suction side of a NACA4412 wing profile, extracted from a direct numerical simulation at Rec = 400,000, is used as the test case. We find that all the methods produce robust results with mild or moderate pressure gradients, but stronger pressure gradients (with β larger than around 7) lead to inconsistent results in all the techniques except the diagnostic plot. This method also has the advantage of providing an objective way of defining the point where the mean streamwise velocity is 99% of the edge velocity, and shows consistent results in a wide range of pressure gradient conditions, as well as flow histories. Therefore, the technique based on the diagnostic plot is a robust method to determine the boundary layer thickness (equivalent to δ99) and edge velocity in pressure gradient turbulent boundary layers.

  6. Free-energy determinants of alpha-helix insertion into lipid bilayers.

    PubMed Central

    Ben-Tal, N; Ben-Shaul, A; Nicholls, A; Honig, B

    1996-01-01

    A detailed treatment is provided of the various free-energy terms that contribute to the transfer of a polyalanine alpha-helix from the aqueous phase into lipid bilayers. In agreement with previous work, the hydrophobic effect is found to provide the major driving force for helix insertion. However, an opposing effect of comparable magnitude is also identified and is attributed to the large free-energy penalty associated with the desolvation of peptide hydrogen bonds on transfer to the low dielectric environment of the bilayer. Lipid perturbation effects as well as the entropy loss associated with helix immobilization in the bilayer are also evaluated. Two configurations of a membrane-bound 25mer polyalanine helix were found to be lower in free energy than the isolated helix in the aqueous phase. The first corresponds to the case of vertical insertion, in which a helix terminus protrudes from each side of the bilayer. The second minimum is for the case of horizontal insertion, for which the helix is adsorbed upon the surface of the bilayer. The calculated free-energy minima are found to be in good agreement with recent measurements of related systems. Large free-energy barriers resulting from desolvation of unsatisfied hydrogen-bonding groups at the helix termini are obtained for both insertion processes. The barriers for insertion are significantly reduced if the helix termini are assumed to be "capped" through the formation of hydrogen bonds with polar sidechains. For uncapped helices, our results support recently proposed models in which helices are inserted by first adsorbing on the membrane surface and then having one terminus "swing around" so as to penetrate the bilayer. Images FIGURE 1 PMID:8785340

  7. Lipid A structure of Pseudoalteromonas haloplanktis TAC 125: use of electrospray ionization tandem mass spectrometry for the determination of fatty acid distribution.

    PubMed

    Corsaro, Maria Michela; Piaz, Fabrizio Dal; Lanzetta, Rosa; Parrilli, Michelangelo

    2002-05-01

    The use of electrospray Ionization (ESI) tandem mass spectrometry (MS/MS) for the structural determination of the lipid A components of the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125 is reported. The lipid A contains the classical bisphosphorylated beta-(1' --> 6)-linked D-glucosamine disaccharide with 3-hydroxydodecanoyl residues (12 : 0 (3-OH)) linked both as esters and amides to 2', 3' (distal glucosamine) and 2, 3 positions (proximal glucosamine) of the sugar backbone. The hydroxyl of 12 : 0 (3-OH) fatty acid linked at the 3' position is esterified by a dodecanoyl residue (12 : 0). In addition to the pentaacyl component, a minor tetraacyl lipid A, lacking the acyl residue at position 3, was also found in the lipid A fraction. The advantage of this MS technique for the investigation of the intra-ring fragmentation, which is useful for the determination of fatty acyl residue distribution on each glucosamine unit, is emphasized.

  8. Computer model of unstirred layer and intracellular pH changes. Determinants of unstirred layer pH.

    PubMed

    Marrannes, Roger

    2013-06-01

    Transmembrane acid-base fluxes affect the intracellular pH and unstirred layer pH around a superfused biological preparation. In this paper the factors influencing the unstirred layer pH and its gradient are studied. An analytical expression of the unstirred layer pH gradient in steady state is derived as a function of simultaneous transmembrane fluxes of (weak) acids and bases with the dehydration reaction of carbonic acid in equilibrium. Also a multicompartment computer model is described consisting of the extracellular bulk compartment, different unstirred layer compartments and the intracellular compartment. With this model also transient changes and the influence of carbonic anhydrase (CA) can be studied. The analytical expression and simulations with the multicompartment model demonstrate that in steady state the unstirred layer pH and its gradient are influenced by the size and type of transmembrane flux of acids and bases, their dissociation constant and diffusion coefficient, the concentration, diffusion coefficient and type of mobile buffers and the activity and location of CA. Similar principles contribute to the amplitude of the unstirred layer pH transients. According to these models an immobile buffer does not influence the steady-state pH, but reduces the amplitude of pH transients especially when these are fast. The unstirred layer pH provides useful information about transmembrane acid-base fluxes. This paper gives more insight how the unstirred layer pH and its transients can be interpreted. Methodological issues are discussed. PMID:23860924

  9. Computer model of unstirred layer and intracellular pH changes. Determinants of unstirred layer pH.

    PubMed

    Marrannes, Roger

    2013-06-01

    Transmembrane acid-base fluxes affect the intracellular pH and unstirred layer pH around a superfused biological preparation. In this paper the factors influencing the unstirred layer pH and its gradient are studied. An analytical expression of the unstirred layer pH gradient in steady state is derived as a function of simultaneous transmembrane fluxes of (weak) acids and bases with the dehydration reaction of carbonic acid in equilibrium. Also a multicompartment computer model is described consisting of the extracellular bulk compartment, different unstirred layer compartments and the intracellular compartment. With this model also transient changes and the influence of carbonic anhydrase (CA) can be studied. The analytical expression and simulations with the multicompartment model demonstrate that in steady state the unstirred layer pH and its gradient are influenced by the size and type of transmembrane flux of acids and bases, their dissociation constant and diffusion coefficient, the concentration, diffusion coefficient and type of mobile buffers and the activity and location of CA. Similar principles contribute to the amplitude of the unstirred layer pH transients. According to these models an immobile buffer does not influence the steady-state pH, but reduces the amplitude of pH transients especially when these are fast. The unstirred layer pH provides useful information about transmembrane acid-base fluxes. This paper gives more insight how the unstirred layer pH and its transients can be interpreted. Methodological issues are discussed.

  10. Thickness of Saturn's ring current determined from north-south Cassini passes through the current layer

    NASA Astrophysics Data System (ADS)

    Kellett, S.; Bunce, E. J.; Coates, A. J.; Cowley, S. W. H.

    2009-04-01

    We investigate magnetic field and plasma electron data from six Cassini orbits during which, unique within the mission to date, the spacecraft passed almost directly north-south through Saturn's equatorial plane within the ring current region of the middle magnetosphere. Three passes took place in the postnoon sector and three took place in the postmidnight sector, each at radial distances of ˜9, ˜12, and ˜15 R S . (R S is Saturn's radius, equal to 60,268 km). These data allow for the first time an essentially direct determination of the north-south field and plasma structure in this region at approximately constant radius and local time. The dayside data indicate the presence of an equatorial current disk with a near constant half-thickness of ˜1.5 R S . Comparison of the magnetic data with simultaneous electron spectrograms shows that the current layer is located within a significantly broader layer of hot plasma. More variable conditions are found on the nightside. In the inner region a thin equatorial current layer of half-thickness ˜0.5 R S is found embedded at the center of a much broader layer of current and plasma. In the central and outer passes the current sheet corresponds to essentially the whole of the hot equatorial electron layer, with a half-thickness of ˜2.5 R S for the central pass, falling strongly to ˜0.4 R S for the outer pass. The data also provide confirmatory evidence of northward displacements of the center of the current layer from Saturn's equatorial plane in the outer regions, on both the dayside and the nightside of the planet.

  11. Density determination of nano-layers depending to the thickness by non-destructive method

    SciTech Connect

    Gacem, A.; Doghmane, A.; Hadjoub, Z.

    2013-12-16

    Non-destructive tests used to characterize and observe the state of the solids near the surface or at depth, without damaging them or damaging them. Density is frequently used to follow the variations of the physical structure of the samples, as well as in the calculation of quantity of material required to fill a given volume, and it is also used to determine the homogeneity of a sample. However, the measurement of the acoustic properties (density, elastic constants,…) of a thin film whose thickness is smaller than several atomic layers is not easy to perform. For that reason, we expose in this work the effects of the thicknesses of thin films on the evolution of the density, where several samples are analyzed. The samples selected structures are thin films deposited on substrates, these coatings have thicknesses varying from a few atomic layers to ten or so micrometers and can change the properties of the substrate on which they are deposited. To do so, we considered a great number of layers (Cr, Al, SiO{sub 2}, ZnO, Cu, AlN, Si{sub 3}N{sub 4}, SiC) deposited on different substrates (Al{sub 2}O{sub 3}, Cu and Quartz). It is first shown that the density exhibits a dispersive behaviour. Such a behaviour is characterized by an initial increase (or decrease) followed by a saturated region. Further investigations of these dependences led to the determination of a semi-empirical universal relations, ρ=f(h/λ{sub T}), for all the investigated layer/substrate combination. Such expression could be of great importance in the density prediction of even layers thicknesses.

  12. Determination of the Schottky barrier height of ferromagnetic contacts to few-layer phosphorene

    SciTech Connect

    Anugrah, Yoska; Robbins, Matthew C.; Koester, Steven J.; Crowell, Paul A.

    2015-03-09

    Phosphorene, the 2D analogue of black phosphorus, is a promising material for studying spin transport due to its low spin-orbit coupling and its ½ nuclear spin, which could allow the study of hyperfine effects. In this work, the properties of permalloy (Py) and cobalt (Co) contacts to few-layer phosphorene are presented. The Schottky barrier height was extracted and determined as a function of gate bias. Flat-band barrier heights, relative to the valence band edge, of 110 meV and 200 meV were determined for Py and Co, respectively. These results are important for future studies of spin transport in phosphorene.

  13. Determination of the Schottky barrier height of ferromagnetic contacts to few-layer phosphorene

    NASA Astrophysics Data System (ADS)

    Anugrah, Yoska; Robbins, Matthew C.; Crowell, Paul A.; Koester, Steven J.

    2015-03-01

    Phosphorene, the 2D analogue of black phosphorus, is a promising material for studying spin transport due to its low spin-orbit coupling and its ½ nuclear spin, which could allow the study of hyperfine effects. In this work, the properties of permalloy (Py) and cobalt (Co) contacts to few-layer phosphorene are presented. The Schottky barrier height was extracted and determined as a function of gate bias. Flat-band barrier heights, relative to the valence band edge, of 110 meV and 200 meV were determined for Py and Co, respectively. These results are important for future studies of spin transport in phosphorene.

  14. Experimental phasing for structure determination using membrane-protein crystals grown by the lipid cubic phase method

    SciTech Connect

    Li, Dianfan; Pye, Valerie E.; Caffrey, Martin

    2015-01-01

    Very little information is available in the literature concerning the experimental heavy-atom phasing of membrane-protein structures where the crystals have been grown using the lipid cubic phase (in meso) method. In this paper, pre-labelling, co-crystallization, soaking, site-specific mercury binding to genetically engineered single-cysteine mutants and selenomethionine labelling as applied to an integral membrane kinase crystallized in meso are described. An assay to assess cysteine accessibility for mercury labelling of membrane proteins is introduced. Despite the marked increase in the number of membrane-protein structures solved using crystals grown by the lipid cubic phase or in meso method, only ten have been determined by SAD/MAD. This is likely to be a consequence of the technical difficulties associated with handling proteins and crystals in the sticky and viscous hosting mesophase that is usually incubated in glass sandwich plates for the purposes of crystallization. Here, a four-year campaign aimed at phasing the in meso structure of the integral membrane diacylglycerol kinase (DgkA) from Escherichia coli is reported. Heavy-atom labelling of this small hydrophobic enzyme was attempted by pre-labelling, co-crystallization, soaking, site-specific mercury binding to genetically engineered single-cysteine mutants and selenomethionine incorporation. Strategies and techniques for special handling are reported, as well as the typical results and the lessons learned for each of these approaches. In addition, an assay to assess the accessibility of cysteine residues in membrane proteins for mercury labelling is introduced. The various techniques and strategies described will provide a valuable reference for future experimental phasing of membrane proteins where crystals are grown by the lipid cubic phase method.

  15. Note: Non-invasive optical method for rapid determination of alignment degree of oriented nanofibrous layers

    NASA Astrophysics Data System (ADS)

    Pokorny, M.; Klemes, J.; Rebicek, J.; Kotzianova, A.; Velebny, V.

    2015-10-01

    This paper presents a rapid non-destructive method that provides information on the anisotropic internal structure of nanofibrous layers. A laser beam of a wavelength of 632.8 nm is directed at and passes through a nanofibrous layer prepared by electrostatic spinning. Information about the structural arrangement of nanofibers in the layer is directly visible in the form of a diffraction image formed on a projection screen or obtained from measured intensities of the laser beam passing through the sample which are determined by the dependency of the angle of the main direction of polarization of the laser beam on the axis of alignment of nanofibers in the sample. Both optical methods were verified on Polyvinyl alcohol (PVA) nanofibrous layers (fiber diameter of 470 nm) with random, single-axis aligned and crossed structures. The obtained results match the results of commonly used methods which apply the analysis of electron microscope images. The presented simple method not only allows samples to be analysed much more rapidly and without damaging them but it also makes possible the analysis of much larger areas, up to several square millimetres, at the same time.

  16. Note: Non-invasive optical method for rapid determination of alignment degree of oriented nanofibrous layers

    SciTech Connect

    Pokorny, M.; Rebicek, J.; Klemes, J.; Kotzianova, A.; Velebny, V.

    2015-10-15

    This paper presents a rapid non-destructive method that provides information on the anisotropic internal structure of nanofibrous layers. A laser beam of a wavelength of 632.8 nm is directed at and passes through a nanofibrous layer prepared by electrostatic spinning. Information about the structural arrangement of nanofibers in the layer is directly visible in the form of a diffraction image formed on a projection screen or obtained from measured intensities of the laser beam passing through the sample which are determined by the dependency of the angle of the main direction of polarization of the laser beam on the axis of alignment of nanofibers in the sample. Both optical methods were verified on Polyvinyl alcohol (PVA) nanofibrous layers (fiber diameter of 470 nm) with random, single-axis aligned and crossed structures. The obtained results match the results of commonly used methods which apply the analysis of electron microscope images. The presented simple method not only allows samples to be analysed much more rapidly and without damaging them but it also makes possible the analysis of much larger areas, up to several square millimetres, at the same time.

  17. Lipid droplets hypertrophy: a crucial determining factor in insulin regulation by adipocytes

    NASA Astrophysics Data System (ADS)

    Sanjabi, Bahram; Dashty, Monireh; Özcan, Behiye; Akbarkhanzadeh, Vishtaseb; Rahimi, Mehran; Vinciguerra, Manlio; van Rooij, Felix; Al-Lahham, Saad; Sheedfar, Fareeba; van Kooten, Theo G.; Spek, C. Arnold; Rowshani, Ajda T.; van der Want, Johannes; Klaassen, Rene; Sijbrands, Eric; Peppelenbosch, Maikel P.; Rezaee, Farhad

    2015-03-01

    Lipid droplets (LDs) hypertrophy in adipocytes is the main cause of energy metabolic system dysfunction, obesity and its afflictions such as T2D. However, the role of adipocytes in linking energy metabolic disorders with insulin regulation is unknown in humans. Human adipocytes constitutively synthesize and secrete insulin, which is biologically functional. Insulin concentrations and release are fat mass- and LDs-dependent respectively. Fat reduction mediated by bariatric surgery repairs obesity-associated T2D. The expression of genes, like PCSK1 (proinsulin conversion enzyme), GCG (Glucagon), GPLD1, CD38 and NNAT, involved in insulin regulation/release were differentially expressed in pancreas and adipose tissue (AT). INS (insulin) and GCG expression reduced in human AT-T2D as compared to AT-control, but remained unchanged in pancreas in either state. Insulin levels (mRNA/protein) were higher in AT derived from prediabetes BB rats with destructed pancreatic β-cells and controls than pancreas derived from the same rats respectively. Insulin expression in 10 human primary cell types including adipocytes and macrophages is an evidence for extrapancreatic insulin-producing cells. The data suggest a crosstalk between AT and pancreas to fine-tune energy metabolic system or may minimize the metabolic damage during diabetes. This study opens new avenues towards T2D therapy with a great impact on public health.

  18. Biochemical determination of lipid content in hepatic steatosis by the Soxtec method

    PubMed Central

    Hijona, Elizabeth; Hijona, Lander; Larzabal, Mikel; Sarasqueta, Cristina; Aldazabal, Pablo; Arenas, Juan; Bujanda, Luis

    2010-01-01

    AIM: To establish a quantitative method to measure the amount of lipids. METHODS: The livers of 53 male Wistar rats (225 g) with different degrees of hepatic steatosis were studied. This model of hepatic steatosis was based on a high carbohydrate, fat-free modified diet. Biopsies were classified into four grades depending on fat accumulation, using the Kleiner and Brunt classification. Total fat was studied by the Soxtec method (Soxtec™ 2050 Auto Fat Extraction System), and agreement between both assays was assessed by calculating the κ coefficient. RESULTS: According to the histological classification, 38% of rats presented grade 0, 21% grade 1, 22% grade 2 and 20% grade 3. The amount of fat per 100 g tissue was 2.60 ± 0.64 g for grade 0, 3.87 ± 1.59 g for grade 1, 5.82 ± 1.37 g for grade 2 and 8.68 ± 2.30 g for grade 3. Statistically significant differences were found between the mean values for each of the histological grades (P < 0.05). The correlation for the quantification of fat in the liver between both assays was moderate (κ = 0.60). CONCLUSION: The biochemical quantification of fat in liver tissue by the Soxtec method was correlated with the histological classification, although the agreement between the two tests was only moderate. PMID:20333790

  19. Experimental phasing for structure determination using membrane-protein crystals grown by the lipid cubic phase method.

    PubMed

    Li, Dianfan; Pye, Valerie E; Caffrey, Martin

    2015-01-01

    Despite the marked increase in the number of membrane-protein structures solved using crystals grown by the lipid cubic phase or in meso method, only ten have been determined by SAD/MAD. This is likely to be a consequence of the technical difficulties associated with handling proteins and crystals in the sticky and viscous hosting mesophase that is usually incubated in glass sandwich plates for the purposes of crystallization. Here, a four-year campaign aimed at phasing the in meso structure of the integral membrane diacylglycerol kinase (DgkA) from Escherichia coli is reported. Heavy-atom labelling of this small hydrophobic enzyme was attempted by pre-labelling, co-crystallization, soaking, site-specific mercury binding to genetically engineered single-cysteine mutants and selenomethionine incorporation. Strategies and techniques for special handling are reported, as well as the typical results and the lessons learned for each of these approaches. In addition, an assay to assess the accessibility of cysteine residues in membrane proteins for mercury labelling is introduced. The various techniques and strategies described will provide a valuable reference for future experimental phasing of membrane proteins where crystals are grown by the lipid cubic phase method.

  20. Experimental phasing for structure determination using membrane-protein crystals grown by the lipid cubic phase method

    PubMed Central

    Li, Dianfan; Pye, Valerie E.; Caffrey, Martin

    2015-01-01

    Despite the marked increase in the number of membrane-protein structures solved using crystals grown by the lipid cubic phase or in meso method, only ten have been determined by SAD/MAD. This is likely to be a consequence of the technical difficulties associated with handling proteins and crystals in the sticky and viscous hosting mesophase that is usually incubated in glass sandwich plates for the purposes of crystallization. Here, a four-year campaign aimed at phasing the in meso structure of the integral membrane diacylglycerol kinase (DgkA) from Escherichia coli is reported. Heavy-atom labelling of this small hydrophobic enzyme was attempted by pre-labelling, co-crystallization, soaking, site-specific mercury binding to genetically engineered single-cysteine mutants and selenomethionine incorporation. Strategies and techniques for special handling are reported, as well as the typical results and the lessons learned for each of these approaches. In addition, an assay to assess the accessibility of cysteine residues in membrane proteins for mercury labelling is introduced. The various techniques and strategies described will provide a valuable reference for future experimental phasing of membrane proteins where crystals are grown by the lipid cubic phase method. PMID:25615865

  1. Determining the Effect of Growth Rate on Hydrogen Isotope Fractionation of Algal Lipids in Two North Pacific Sites

    NASA Astrophysics Data System (ADS)

    Wolfshorndl, M.; Sachs, J. P.

    2015-12-01

    Tropical hydrologic changes have a large effect on global climate, but there does not yet exist a good indicator of rainfall variation in the tropics. Understanding past natural variability of such features as the Intertropical Convergence Zone and El Niño Southern Oscillation provides information about the extent of anthropogenic climate change today. The hydrogen isotopic composition (D/H ratio) of algal lipids has been shown to track the isotopic composition of source water in which the organism grew, providing information about precipitation variability over time. However, culture work has revealed that environmental factors such as salinity, temperature, growth rate, and irradiance also influence algal lipid D/H ratios. Here I present work determining the effect of growth rate and irradiance on the hydrogen isotope composition of alkenone-producing algae in the water column in two North Pacific locations, off the coast of Oregon and near the Hawaii Ocean Time Series site. This work corroborates empirical relationships observed in culture studies and indicates that the effects of growth rate and irradiance should be taken into account when applying the D/H isotope ratio rainfall proxy to reconstruct past climates.

  2. Density of states determination in organic donor-acceptor blend layers enabled by molecular doping

    NASA Astrophysics Data System (ADS)

    Fischer, Janine; Ray, Debdutta; Kleemann, Hans; Pahner, Paul; Schwarze, Martin; Koerner, Christian; Vandewal, Koen; Leo, Karl

    2015-06-01

    Charge carrier transport is a key parameter determining the efficiency of organic solar cells, and is closely related to the density of free and trapped states. For trap characterization, impedance spectroscopy is a suitable, non-invasive method, applicable to complete organic semiconductor devices. In order to contribute to the capacitive signal, the traps must be filled with charge carriers. Typically, trap filling is achieved by illuminating the device or by injecting charge carriers through application of a forward bias voltage. However, in both cases, the exact number of charge carriers in the device is not known and depends strongly on the measurement conditions. Here, hole trap states of the model blend layer ZnPc:C60 are filled by weak p-doping, enabling trap characterization in a blend layer at a controlled hole density. We evaluate impedance spectra at different temperatures in order to determine the density of occupied states (DOOS) directly from the capacitance-frequency spectra by assuming a simple energy diagram. The reconstructed DOOS distribution is analyzed at different doping concentrations and device thicknesses and compared to thermally stimulated current measurements performed on the same devices. In both methods, a pronounced Gaussian peak at about 0.4 eV below the transport level is found as well as deep, exponential tail states, providing a deeper insight into the density of states distribution of this donor-acceptor blend layer. Additionally, the effect of doping-induced trap filling on the solar cell characteristics is studied in these devices.

  3. Parenteral lipid fatty acid composition directly determines the fatty acid composition of red blood cell and brain lipids in preterm pigs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Studies in enterally-fed infants have shown a positive effect of n-3 long-chain polyunsaturated fatty acid (LCPUFA) supplementatin on neurodevelopment. The effect of n-3 LCPUFA in fish oil-based parenteral (PN) lipid emulsions on neuronal tissues of PN-fed preterm infants is unknown. The objective ...

  4. Simultaneous determination of water-soluble vitamins by over-pressure layer chromatography and photodensitometric detection.

    PubMed

    Postaire, E; Cisse, M; Le Hoang, M D; Pradeau, D

    1991-04-01

    An over-pressure layer chromatographic procedure with photodensitometric detection for the simultaneous determination of water-soluble vitamins in multivitamin pharmaceutical preparations was developed and evaluated. The method uses high-performance TLC (HPTLC) plates with silica gel as the thin-layer, and an n-butanol:pyridine:water mixture (50:35:15, v/v/v) as mobile phase at a rate of 0.25 mL/min for baseline separation. The quantitation was carried out without derivatization (vitamin B1, vitamin B2, vitamin B6, folic acid, nicotinamide, vitamin C) or after spraying ninhydrin reagent (calcium pantothenate) or 4-dimethylaminocinnamaldehyde (vitamin B12, biotin). This was applied to the analysis of multivitamin solutions. Satisfactory relative standard deviations and good recovery were obtained for all the vitamins examined. It was concluded that this method is fast, accurate, specific, and suitable for routine quality control use. PMID:1865338

  5. The role of a small-scale cutoff in determining molecular layers at fluid interfaces.

    PubMed

    Sega, Marcello

    2016-08-17

    The existence of molecular layers at liquid/vapour interfaces has been a long debated issue. More than ten years ago it was shown, using computer simulations, that correlations at the liquid/vapour interface resemble those of bulk liquids, even though they can be detected in experiments only in a few cases, where they are so strong that they cannot be concealed by the geometrical smearing of capillary fluctuations. The results of the intrinsic analysis techniques used in computer experiments, however, are still often questioned because of their dependence on a free parameter that usually represents a small-scale cutoff used to determine the interface. In this work I show that there is only one value of the cutoff that can ensure a quantitative explanation of the intrinsic density correlation peaks in terms of successive layer contributions. The value of the cutoff coincides, with a high accuracy, with the molecular diameter. PMID:27499039

  6. Determination of isotropic layer parameters from spatiotemporal signals of an ultrasonic array

    NASA Astrophysics Data System (ADS)

    Titov, S. A.; Maev, R. G.

    2013-09-01

    The paper discusses a method for measuring the velocities and attenuation of longitudinal and transverse ultrasonic waves and the density and thickness of the isotropic layer with an array placed in an immersion liquid parallel to the sample. The method is based on the recording of the total spatiotemporal signal of the array and its expansion into a spatial spectrum of pulse plane wave response. The ultrasonic velocity and sample thickness depend on the response delay of the plane wave in the layer from the transverse projection of the slowness vector. The density and attenuation are determined from the behavior of the amplitudes of spectral responses. To confirm this method in experiment, the parameters of a polystyrene plate have been measured using a linear 32-element array with a central frequency of 17 MHz.

  7. On determining characteristic length scales in pressure-gradient turbulent boundary layers

    NASA Astrophysics Data System (ADS)

    Vinuesa, R.; Bobke, A.; Örlü, R.; Schlatter, P.

    2016-05-01

    In the present work, we analyze three commonly used methods to determine the edge of pressure gradient turbulent boundary layers: two based on composite profiles, the one by Chauhan et al. ["Criteria for assessing experiments in zero pressure gradient boundary layers," Fluid Dyn. Res. 41, 021404 (2009)] and the one by Nickels ["Inner scaling for wall-bounded flows subject to large pressure gradients," J. Fluid Mech. 521, 217-239 (2004)], and the other one based on the condition of vanishing mean velocity gradient. Additionally, a new method is introduced based on the diagnostic plot concept by Alfredsson et al. ["A new scaling for the streamwise turbulence intensity in wall-bounded turbulent flows and what it tells us about the `outer' peak," Phys. Fluids 23, 041702 (2011)]. The boundary layers developing over the suction and pressure sides of a NACA4412 wing section, extracted from a direct numerical simulation at chord Reynolds number Rec = 400 000, are used as the test case, besides other numerical and experimental data from favorable, zero, and adverse pressure-gradient flat-plate turbulent boundary layers. We find that all the methods produce robust results with mild or moderate pressure gradients, although the composite-profile techniques require data preparation, including initial estimations of fitting parameters and data truncation. Stronger pressure gradients (with a Rotta-Clauser pressure-gradient parameter β larger than around 7) lead to inconsistent results in all the techniques except the diagnostic plot. This method also has the advantage of providing an objective way of defining the point where the mean streamwise velocity is 99% of the edge velocity and shows consistent results in a wide range of pressure gradient conditions, as well as flow histories. Collapse of intermittency factors obtained from a wide range of pressure-gradient and Re conditions on the wing further highlights the robustness of the diagnostic plot method to determine the

  8. Determination of band alignment in the single-layer MoS2/WSe2 heterojunction

    PubMed Central

    Chiu, Ming-Hui; Zhang, Chendong; Shiu, Hung-Wei; Chuu, Chih-Piao; Chen, Chang-Hsiao; Chang, Chih-Yuan S.; Chen, Chia-Hao; Chou, Mei-Yin; Shih, Chih-Kang; Li, Lain-Jong

    2015-01-01

    The emergence of two-dimensional electronic materials has stimulated proposals of novel electronic and photonic devices based on the heterostructures of transition metal dichalcogenides. Here we report the determination of band offsets in the heterostructures of transition metal dichalcogenides by using microbeam X-ray photoelectron spectroscopy and scanning tunnelling microscopy/spectroscopy. We determine a type-II alignment between MoS2 and WSe2 with a valence band offset value of 0.83 eV and a conduction band offset of 0.76 eV. First-principles calculations show that in this heterostructure with dissimilar chalcogen atoms, the electronic structures of WSe2 and MoS2 are well retained in their respective layers due to a weak interlayer coupling. Moreover, a valence band offset of 0.94 eV is obtained from density functional theory, consistent with the experimental determination. PMID:26179885

  9. Determination of band alignment in the single-layer MoS2/WSe2 heterojunction.

    PubMed

    Chiu, Ming-Hui; Zhang, Chendong; Shiu, Hung-Wei; Chuu, Chih-Piao; Chen, Chang-Hsiao; Chang, Chih-Yuan S; Chen, Chia-Hao; Chou, Mei-Yin; Shih, Chih-Kang; Li, Lain-Jong

    2015-07-16

    The emergence of two-dimensional electronic materials has stimulated proposals of novel electronic and photonic devices based on the heterostructures of transition metal dichalcogenides. Here we report the determination of band offsets in the heterostructures of transition metal dichalcogenides by using microbeam X-ray photoelectron spectroscopy and scanning tunnelling microscopy/spectroscopy. We determine a type-II alignment between MoS2 and WSe2 with a valence band offset value of 0.83 eV and a conduction band offset of 0.76 eV. First-principles calculations show that in this heterostructure with dissimilar chalcogen atoms, the electronic structures of WSe2 and MoS2 are well retained in their respective layers due to a weak interlayer coupling. Moreover, a valence band offset of 0.94 eV is obtained from density functional theory, consistent with the experimental determination.

  10. To determine ice layer thickness of Europa by high energy neutrino

    NASA Astrophysics Data System (ADS)

    Shoji, D.; Kurita, K.; Tanaka, H. K.

    2010-12-01

    Europa, the second closest Galilean satellite is one of the targets which are suspected to have an internal ocean. Detection and characterization of the internal ocean is one of the main subjects for Europa orbiter exploration. Although the gravitational data has shown the thickness of the surface H2O layer of 80-170km[1], it can not determine the phase of H2O. The variations in the magnetic field associated with the induced current in the internal ocean can determine the thickness of the layer of ice if satellite's orbits satisfy the required conditions. Observations of tidal amplitude forced by Jupiter can also resolve the thickness of the surface lithosphere[2]. At moment because of the lack of observational constraints there exist two contrasting models:thick ice layer model and thin model. Here we propose new method to detect the ocean directly based on the radiation by high energy neutrino interacted with matter. Schaefer et al[3] have proposed a similar method to determine ice layer thickness. We will focus on the detection of internal ocean for Europa and present the method is suitable for actual situations of Europa exploration by numerical simulations. Neutrino is famous for its traveling at long distance without any interaction with matter. When high energy neutrinos traverse in Europa hadronic showers are produced by the weak interaction with the nucleons that makes the body of Europa. These hadronic showers induces excess electrons. Because of these excess electrons, Cherenkov photons are emitted. When this radiation occurs in the ice layer, radiations whose wave length is over 10cm should be coherent because the scale of the shower becomes small (a few cm) in the ice, which is called as Askaryan effect[3]. Thus, the intensity of the radiation whose frequency is a few GHz should be enhanced. Since ice has a much longer attenuation length than water, the radiations which occur in the surface ice layer could be detected by the antenna outside Europa but

  11. Novel therapeutic mechanisms determine the effectiveness of lipid-core nanocapsules on melanoma models

    PubMed Central

    Drewes, Carine C; Fiel, Luana A; Bexiga, Celina G; Asbahr, Ana Carolina C; Uchiyama, Mayara K; Cogliati, Bruno; Araki, Koiti; Guterres, Sílvia S; Pohlmann, Adriana R; Farsky, Sandra P

    2016-01-01

    Melanoma is a severe metastatic skin cancer with poor prognosis and no effective treatment. Therefore, novel therapeutic approaches using nanotechnology have been proposed to improve therapeutic effectiveness. Lipid-core nanocapsules (LNCs), prepared with poly(ε-caprolactone), capric/caprylic triglyceride, and sorbitan monostearate and stabilized by polysorbate 80, are efficient as drug delivery systems. Here, we investigated the effects of acetyleugenol-loaded LNC (AcE-LNC) on human SK-Mel-28 melanoma cells and its therapeutic efficacies on melanoma induced by B16F10 in C57B6 mice. LNC and AcE-LNC had z-average diameters and zeta potential close to 210 nm and -10.0 mV, respectively. CytoViva® microscopy images showed that LNC and AcE-LNC penetrated into SK-Mel-28 cells, and remained in the cytoplasm. AcE-LNC in vitro treatment (18–90×109 particles/mL; 1 hour) induced late apoptosis and necrosis; LNC and AcE-LNC (3–18×109 particles/mL; 48 hours) treatments reduced cell proliferation and delayed the cell cycle. Elevated levels of nitric oxide were found in supernatant of LNC and AcE-LNC, which were not dependent on nitric oxide synthase expressions. Daily intraperitoneal or oral treatment (days 3–10 after tumor injection) with LNC or AcE-LNC (1×1012 particles/day), but not with AcE (50 mg/kg/day, same dose as AcE-LNC), reduced the volume of the tumor; nevertheless, intraperitoneal treatment caused toxicity. Oral LNC treatment was more efficient than AcE-LNC treatment. Moreover, oral treatment with nonencapsulated capric/caprylic triglyceride did not inhibit tumor development, implying that nanocapsule supramolecular structure is important to the therapeutic effects. Together, data herein presented highlight the relevance of the supramolecular structure of LNCs to toxicity on SK-Mel-28 cells and to the therapeutic efficacy on melanoma development in mice, conferring novel therapeutic mechanisms to LNC further than a drug delivery system. PMID:27099491

  12. Determination of lawsone in henna powders by high performance thin layer chromatography.

    PubMed

    El-Shaer, Nagwa S; Badr, Jihan M; Aboul-Ela, Maha A; Gohar, Yousry M

    2007-12-01

    The lawsone content has been evaluated quantitatively in eight commercial henna powders and two collected henna leaves. The phenolic, chloroform-soluble fraction of the majority of the examined samples showed the presence of lawsone and two other pigments. Here we aimed to optimize high performance thin layer chromatography for the determination of lawsone. Upon using the optimized method the examined samples showed considerable variation in lawsone concentration ranging from 0.004 up to 0.608 wt%, indicating that some samples were almost devoid of lawsone. Some of these products were subjected to preliminary in vivo toxicity studies.

  13. Determination by spaceborne backscatter lidar of the structural parameters of atmospheric scattering layers.

    PubMed

    Chazette, P; Pelon, J; Mégie, G

    2001-07-20

    Spaceborne active lidar systems are under development to give new insight into the vertical distribution of clouds and aerosols in the atmosphere and to provide new information on variables required for improvement of forecast models and for understanding the radiative and dynamic processes that are linked to the dynamics of climate change. However, when they are operated from space, lidar systems are limited by atmospheric backscattered signals that have low signal-to-noise ratios (SNRs) on optically thin targets. Therefore specific methods of analysis have to be developed to ensure accurate determination of the geometric and optical properties of scattering layers in the atmosphere. A first approach to retrieving the geometric properties of semitransparent cloud and aerosol layers is presented as a function of false-alarm and no-detection probabilities for a given SNR. Simulations show that the geometric properties of thin cirrus clouds and the altitude of the top of the unstable atmospheric boundary layer can be retrieved with standard deviations smaller than 150 m for a vertical resolution of the lidar system in the 50-100-m range and a SNR of 3. The altitudes of the top of dense clouds are retrieved with a precision in altitude of better than 50 m, as this retrieval corresponds to a higher SNR value. Such methods have an important potential application to future spaceborne lidar missions.

  14. Shielding application of perturbation theory to determine changes in neutron and gamma doses due to changes in shield layers

    NASA Technical Reports Server (NTRS)

    Fieno, D.

    1972-01-01

    Perturbation theory formulas were derived and applied to determine changes in neutron and gamma-ray doses due to changes in various radiation shield layers for fixed sources. For a given source and detector position, the perturbation method enables dose derivatives with respect to density, or equivalently thickness, for every layer to be determined from one forward and one inhomogeneous adjoint calculation. A direct determination without the perturbation approach would require two forward calculations to evaluate the dose derivative due to a change in a single layer. Hence, the perturbation method for obtaining dose derivatives requires fewer computations for design studies of multilayer shields. For an illustrative problem, a comparison was made of the fractional change in the dose per unit change in the thickness of each shield layer in a two-layer spherical configuration as calculated by perturbation theory and by successive direct calculations; excellent agreement was obtained between the two methods.

  15. Bioelectronic tongue based on lipidic nanostructured layers containing phenol oxidases and lutetium bisphthalocyanine for the analysis of grapes.

    PubMed

    Medina-Plaza, C; de Saja, J A; Rodriguez-Mendez, M L

    2014-07-15

    In this work, a multisensor system formed by nanostructured voltammetric biosensors based on phenol oxidases (tyrosinase and laccase) has been developed. The enzymes have been incorporated into a biomimetic environment provided by a Langmuir-Blodgett (LB) film of arachidic acid (AA). Lutetium bisphthalocyanine (LuPc2) has also been introduced in the films to act as electron mediator. The incorporation of the enzymes to the floating layers to form Tyr/AA/LuPc2 and Lac/AA/LuPc2 films has been confirmed by the expansion in the surface pressure isotherms and by the AFM images. The voltammetric response towards six phenolic compounds demonstrates the enhanced performance of the biosensors that resulted from a preserved activity of the tyrosinase and laccase combined with the electron transfer activity of LuPc2. Biosensors show improved detection limits in the range of 10(-7)-10(-8) mol L(-1). An array formed by three sensors AA/LuPc2, Tyr/AA/LuPc2 and Lac/AA/LuPc2 has been employed to discriminate phenolic antioxidants of interest in the food industry. The Principal Component Analysis scores plot has demonstrated that the multisensor system is able to discriminate phenols according to the number of phenolic groups attached to the structure. The system has also been able to discriminate grapes of different varieties according to their phenolic content. This good performance is due to the combination of four factors: the high functionality of the enzyme obtained using a biomimetic immobilization, the signal enhancement caused by the LuPc2 mediator, the improvement in the selectivity induced by the enzymes and the complementary activity of the enzymatic sensors demonstrated in the loading plots.

  16. Determination of bergapten in fragrance preparations by thin layer chromatography and spectrophotofluorometry.

    PubMed

    Wisneski, H H

    1976-05-01

    A method has been developed for the determination of bergapten (5-methoxypsoralen), a known phototoxin, in perfumes,colognes, and toilet waters. The bergapten and other lactonic compounds were first isolated from the sample by a series of extractions. The extract containing the bergapten was diluted to a known volume and a aliquot was spotted on a thin layer chromatographic (TLC) plate coated with silica gel G. After 2-dimensional development with hexane - carbon tetrachloride - tert - butylamine (180+12+9) and hexane-toluene-ethyl acetate-acetic acid (100+10+15+0.5), the TLC plate was dried and the emitted fluorescence of bergapten was measured, using a spectrophotofluorometer equipped with a TLC accessory and coupled to an automatic digital integrator. The amount of bergapten was determined by comparing its peak area to those of bergapten standards. The average recovery for levels of 0.001, 0.005, and 0.01% bergapten was 88%.

  17. Highly sensitive determination of chloramphenicol based on thin-layered MoS2/polyaniline nanocomposite.

    PubMed

    Yang, Tao; Chen, Huaiyin; Ge, Tong; Wang, Jin; Li, Weihua; Jiao, Kui

    2015-11-01

    The nanocomposite of molybdenum disulfide (MoS2) and polyaniline (PANI) was prepared through in situ polymerization of aniline on the surface and interlayer of thin-layered MoS2. Owing to the physisorption of aromatic aniline onto the basal plane of MoS2, the electrochemical properties of MoS2/PANI nanocomposite were improved. And a novel electrochemical sensor based on MoS2/PANI nanocomposite was used to determine chloramphenicol by differential pulse voltammetry, exhibiting excellent performance. The detection range was from 1×10(-7) mol L(-1) to 1×10(-4) mol L(-1), with a high sensitivity and a low detection limit of 6.9×10(-8) mol L(-1). In addition, this sensor can be used for the determination of chloramphenicol in real samples. PMID:26452965

  18. Highly sensitive determination of chloramphenicol based on thin-layered MoS2/polyaniline nanocomposite.

    PubMed

    Yang, Tao; Chen, Huaiyin; Ge, Tong; Wang, Jin; Li, Weihua; Jiao, Kui

    2015-11-01

    The nanocomposite of molybdenum disulfide (MoS2) and polyaniline (PANI) was prepared through in situ polymerization of aniline on the surface and interlayer of thin-layered MoS2. Owing to the physisorption of aromatic aniline onto the basal plane of MoS2, the electrochemical properties of MoS2/PANI nanocomposite were improved. And a novel electrochemical sensor based on MoS2/PANI nanocomposite was used to determine chloramphenicol by differential pulse voltammetry, exhibiting excellent performance. The detection range was from 1×10(-7) mol L(-1) to 1×10(-4) mol L(-1), with a high sensitivity and a low detection limit of 6.9×10(-8) mol L(-1). In addition, this sensor can be used for the determination of chloramphenicol in real samples.

  19. Selective separation and determination of isoproterenol on thin layers of bismuth silicate ion-exchanger.

    PubMed

    Ghoulipour Vanik; Hassankhani-Majd Zahra

    2015-06-01

    A simple and sensitive method for the separation and determination of isoproterenol from other doping drugs has been developed on thin layers of bismuth silicate, a synthetic inorganic ion exchanger as adsorbent in thin layer chromatography (TLC). A mixture of methanol and 0.1 mol/L formic acid (3:7, v/v) was employed as the mobile phase. The development time was 32 min. The quantitative measurement were performed with a Camag TLC Scanner-3 at wavelength (λ) of 410 nm. The isoproterenol recovery in this procedure was 98.9%. The linear correlation coefficient was greater than 0. 987 1 and the relative standard deviation (RSD) was less than 0.94. The limit of detection (LOD) and limit of quantification ( LOQ) were 7.7 x 10(-7) mol/L and 3.85 x 10(-6) mol/L, respectively. This method has been applied in the determination of isoproterenol in dosage forms and in biological fluids.

  20. Determination and comparison of how the chain number and chain length of a lipid affects its interactions with a phospholipid at an air/water interface.

    PubMed

    Ngyugen, Hang; McNamee, Cathy E

    2014-06-01

    We determined how the number of chains in a lipid and its chain length affects its interactions with a phospholipid model membrane, and whether the number of chains or the chain length of lipids affects their interactions with the phospholipids more. This was achieved by using a Langmuir trough and a fluorescence microscope to study the interactions of mono-, di-, and triglycerides with a phospholipid monolayer at an air/water interface. The effect of the number of chains in a lipid on its interactions with phospholipids at air/water interfaces was shown by surface pressure-area per molecule isotherms and their thermodynamic analysis to worsen as the number of alkyl chains was increased to be greater than one. An increase in the packing density decreased the mixing ability of the lipids with the phospholipids, resulting in the formation of aggregates in the mixed monolayer. The aggregation was explained by the intermolecular hydrophobic and van der Waals attractions between the lipid molecules. Fluorescence microscopy revealed partial mixing without aggregation for monoglycerides, but the presence of lipid aggregation for diglycerides and triglycerides. The effect of decreasing the chain length of triglycerides from a long chain to a medium chain caused the interactions of the lipids with the phospholipid molecules at the air/water interface to significantly improve. Decreasing the chain length of monoglycerides from a long chain to a medium chain worsened their interaction with the phospholipid molecules. The effect of decreasing the triglyceride chain length on their interactions with phospholipids was much greater than the effect of decreasing the number of alkyl chains in the lipid.

  1. [Stability and flexibility of the lipid layer of the tear film and their pathologic changes in a biophysical experiment].

    PubMed

    Kaercher, T; Möbius, D

    1989-01-01

    In vitro experiments with meibomian gland secretion from healthy subjects, patients with keratoconjunctivitis sicca and meibomianitis were performed. In addition, patients of different age groups of both sexes were also tested. The biophysical experiments included continuous measurement of surface pressure and surface potential under compression and decompression conditions. Surface pressure was determined by a Wilhelmy type film balance; surface potential was determined using the vibrating plate method. In the age group up to 60 years, the surface potential of females, males and patients with keratoconjunctivitis sicca showed a characteristic peak of 200 mV under compression, which was fully reversible under decompression. Females, males and patients with keratoconjunctivitis sicca over 60 years of age showed changes in the surface potential under decompression which indicated that the secretion was unstable and inhomogeneons. Patients with meibomianitis presented characteristic changes of surface potential. Qualitative alteration of the secretion may be the corresponding interpretation. The above biophysical methods have been used for the first time under clinical aspects.

  2. Determination of lipid and phenolic fraction in two hazelnut (Corylus avellana L.) cultivars grown in Poland.

    PubMed

    Ciemniewska-Żytkiewicz, Hanna; Verardo, Vito; Pasini, Federica; Bryś, Joanna; Koczoń, Piotr; Caboni, Maria Fiorenza

    2015-02-01

    The fatty acid, tocopherol, sterol, phospholipid and phenolic compositions of Polish hazelnuts (Kataloński and Webba Cenny) were examined. Particularly, free+esterified and bound tocopherol, sterol and phenolic compounds were determined. The major fatty acids found in hazelnuts were oleic and linoleic acids. α-Tocopherol was the most abundant tocopherol accounting for 90-92% of the total content. Bound tocopherols represented 45.5% and 21.7% of total tocopherols in Kataloński and Webba Cenny cultivar, respectively. Total free+esterified sterols were between 62.0% and 75.7% of total sterols and β-sitosterol was the first sterol in the two samples. Phosphatidylcholine was the most common phospholipid, accounting for 72.2% for Kataloński and 67.5% Webba Cenny, respectively. The most abundant fatty acids in the phospholipid fraction were oleic equally with palmitic acids. Twelve free and six bound phenolic compounds were identified and quantified in hazelnut kernel, instead nine free and six bound phenolic compounds were determined in hard shell.

  3. Determination of nitrogen to carbon abundance ratios from transition layer emission lines

    NASA Technical Reports Server (NTRS)

    Boehm-Vitense, Erika

    1992-01-01

    We have finished studying the nitrogen to carbon abundance ratios for stars with different effective temperatures T(sub eff) and luminosities using transition layer emission lines and using spectra available in the IUE archives. The N/C abundance ratio determinations using transition layer emission lines are as accurate as the photospheric abundance determinations as found by comparison of results obtained by both methods for the same stars. Our measurements confirm photospheric abundance determinations in regions of the HR diagram where they can be obtained. Our studies have extended the temperature range to higher temperatures. They have shown the exact positions in the HR diagram where the mixing due to the outer convection zones reaches deep enough to bring nuclear processed material to the surface. This occurs at effective temperatures which are higher by delta log T(sub eff) approximately 0.04 or roughly 400 K than expected theoretically. Since the depth of the convection zone increases rapidly with decreasing T(sub eff) this may indicate considerable overshoot beyond the lower boundary of the convection zone. Our N/C abundance ratio determinations from transition layer emission lines have confirmed that the actual enrichment observed for some cool giants is larger than expected theoretically, again indicating a larger degree of mixing in several stars either from below or from above. For the supergiants it probably indicates overshoot above the convective core in the progenitor main sequence stars. For the more massive giants this may also be the case, though we did not find a correlation between delta log N/C and the absolute magnitudes, but these are rather uncertain. As byproducts of these studies we also found anomalies in Si/C and N/C abundance ratios for F giants which can be understood as the relict of surface abundance changes for their main sequence progenitors due to diffusion. This anomaly disappears for G giants, for which the depths of the

  4. The dynamics of the plasmasphere boundary layer as determined by ground magnetometers, satellite observations, and modeling

    NASA Astrophysics Data System (ADS)

    Zesta, E.; Boudouridis, A.; Yizengaw, E.; Jorgensen, A. M.; Carranza-fulmer, T. L.; Moldwin, M.; Mann, I. R.; Chi, P. J.

    2013-12-01

    The plasmasphere boundary layer (PBL) separates the cold and dense plasmaspheric plasma from the more tenuous and hot plasma sheet plasma and organizes the spatial distribution of ULF and VLF waves that can contribute to acceleration or loss processes of radiation belt particles through wave-particle interactions. The PBL has been traditionally determined by in situ observations and can be given by empirical models. Recent work has shown that a mid-latitude chain of well-spaced ground magnetometers can also determine the PBL location. Spectral properties, like the cross-phase reversal in the standard field-line resonance (FLR) determination between two stations closely aligned in latitude, have been shown to indicate the presence of a sharp PBL. We merge data from many ground magnetometer pairs from the SAMBA (South American Meridional B-field Array), McMAC (Mid continent Magnetoseismic Chain), and CARISMA (Canadian Array for Realtime Investigations of Magnetic Activity) chains to provide the best available spatial coverage in L values spanning the plasmasphere and PBL, for a range of dynamic states (L=1.6 to greater than 5). The PBL location is identified as the L value of the station pair for which a reverse phase difference is observed in the standard FLR determination. We compare the FLR determined PBL with the trough boundary determined by GPS Total Electron Content (TEC) analysis and with model PBL. Initial results demonstrate that the PBL as identified by the reverse phase differences is in general agreement with TEC identifications and reasonable agreement with models. Reverse Phase Differences are regularly observed at the L range of 2.7 to 3.7 and are highly correlated with Dst and Kp, as determined by daily correlations. We further examine the more detailed time sequence of the PBL dynamics by focusing on key periods around storms preceded by quiet periods, and by using the full L range of the ground magnetometer pairs.

  5. Dual-Layered Nanogel-Coated Hollow Lipid/Polypeptide Conjugate Assemblies for Potential pH-Triggered Intracellular Drug Release

    PubMed Central

    Chiang, Wen-Hsuan; Huang, Wen-Chia; Shen, Ming-Yin; Wang, Che-Hsu; Huang, Yi-Fong; Lin, Sung-Chyr; Chern, Chorng-Shyan; Chiu, Hsin-Cheng

    2014-01-01

    To achieve effective intracellular anticancer drug delivery, the polymeric vesicles supplemented with the pH-responsive outlayered gels as a delivery system of doxorubicin (DOX) were developed from self-assembly of the lipid/polypeptide adduct, distearin grafted poly(γ-glutamic acid) (poly(γ-GA)), followed by sequential deposition of chitosan and poly(γ-GA-co-γ-glutamyl oxysuccinimide)-g-monomethoxy poly(ethylene glycol) in combination with in situ covalent cross-linking on assembly surfaces. The resultant gel-caged polymeric vesicles (GCPVs) showed superior performance in regulating drug release in response to the external pH change. Under typical physiological conditions (pH 7.4 and 37°C) at which the γ-GA/DOX ionic pairings remained mostly undisturbed, the dense outlayered gels of GCPVs significantly reduced the premature leakage of the uncomplexed payload. With the environmental pH being reduced from pH 7.4 to 4.7, the drug liberation was appreciably promoted by the massive disruption of the ionic γ-GA/DOX complexes along with the significant swelling of nanogel layers upon the increased protonation of chitosan chain segments. After being internalized by HeLa cells via endocytosis, GCPVs exhibited cytotoxic effect comparable to free DOX achieved by rapidly releasing the payload in intracellular acidic endosomes and lysosomes. This strongly implies the great promise of such unique GCPVs as an intracellular drug delivery carrier for potential anticancer treatment. PMID:24651156

  6. An octaethylene glycol monododecyl ether-based mixed micellar assay for determining the lipid acyl hydrolase activity of patatin.

    PubMed

    Jiménez, M; Escribano, J; Pérez-Gilabert, M; Chazarra, S; Cabanes, J; García-Carmona, F

    2001-10-01

    Patatin was extracted from potato tubers (Solanum tuberosum L. cv. Spunta) and purified to homogeneity by ammonium sulfate salt fractionation and one sole chromatographic step. A spectrophotometric mixed micellar assay for patatin lipid acyl hydrolase (LAH) activity was designed with the detergent octaethylene glycol monododecyl ether (C12E8). Patatin LAH used p-nitrophenyl butyrate (PNP-butyrate) as substrate when solubilized in (C12E8) micelles. In the mixed micellar system, patatin LAH responds to the PNP-butyrate surface concentration expressed as mol% (= [PNP-butyratel x 100/([detergentl critical micellar concentration)) and not to the molarity of PNP-butyrate. The kinetic parameters were determined; Vmax was independent of the mixed micelle concentration, as was Km, when expressed as mol%. However, Km was dependent on C12E8 concentration when expressed in molar concentration. C12E8/PNP-butyrate proved to be a reliable system for assaying patatin LAH activity and is superior to the commonly used Triton X-100 and SDS methods. It permits investigation of the substrate requirements of patatin LAH activity because the concentration-independent Km can be determined both in mol% and as the absolute number of substrate molecules per micelle. In addition, the detergent did not affect the enzyme activity.

  7. Competing Lipid-Protein and Protein-Protein Interactions Determine Clustering and Gating Patterns in the Potassium Channel from Streptomyces lividans (KcsA)*

    PubMed Central

    Molina, M. Luisa; Giudici, A. Marcela; Poveda, José A.; Fernández-Ballester, Gregorio; Montoya, Estefanía; Renart, M. Lourdes; Fernández, Asia M.; Encinar, José A.; Riquelme, Gloria; Morales, Andrés; González-Ros, José M.

    2015-01-01

    There is increasing evidence to support the notion that membrane proteins, instead of being isolated components floating in a fluid lipid environment, can be assembled into supramolecular complexes that take part in a variety of cooperative cellular functions. The interplay between lipid-protein and protein-protein interactions is expected to be a determinant factor in the assembly and dynamics of such membrane complexes. Here we report on a role of anionic phospholipids in determining the extent of clustering of KcsA, a model potassium channel. Assembly/disassembly of channel clusters occurs, at least partly, as a consequence of competing lipid-protein and protein-protein interactions at nonannular lipid binding sites on the channel surface and brings about profound changes in the gating properties of the channel. Our results suggest that these latter effects of anionic lipids are mediated via the Trp67–Glu71–Asp80 inactivation triad within the channel structure and its bearing on the selectivity filter. PMID:26336105

  8. Competing Lipid-Protein and Protein-Protein Interactions Determine Clustering and Gating Patterns in the Potassium Channel from Streptomyces lividans (KcsA).

    PubMed

    Molina, M Luisa; Giudici, A Marcela; Poveda, José A; Fernández-Ballester, Gregorio; Montoya, Estefanía; Renart, M Lourdes; Fernández, Asia M; Encinar, José A; Riquelme, Gloria; Morales, Andrés; González-Ros, José M

    2015-10-16

    There is increasing evidence to support the notion that membrane proteins, instead of being isolated components floating in a fluid lipid environment, can be assembled into supramolecular complexes that take part in a variety of cooperative cellular functions. The interplay between lipid-protein and protein-protein interactions is expected to be a determinant factor in the assembly and dynamics of such membrane complexes. Here we report on a role of anionic phospholipids in determining the extent of clustering of KcsA, a model potassium channel. Assembly/disassembly of channel clusters occurs, at least partly, as a consequence of competing lipid-protein and protein-protein interactions at nonannular lipid binding sites on the channel surface and brings about profound changes in the gating properties of the channel. Our results suggest that these latter effects of anionic lipids are mediated via the Trp(67)-Glu(71)-Asp(80) inactivation triad within the channel structure and its bearing on the selectivity filter.

  9. Determination of Total Lipids as Fatty Acid Methyl Esters (FAME) by in situ Transesterification: Laboratory Analytical Procedure (LAP)

    SciTech Connect

    Van Wychen, S.; Laurens, L. M. L.

    2013-12-01

    This procedure is based on a whole biomass transesterification of lipids to fatty acid methyl esters to represent an accurate reflection of the potential of microalgal biofuels. Lipids are present in many forms and play various roles within an algal cell, from cell membrane phospholipids to energy stored as triacylglycerols.

  10. Mixing layer height measurements determines influence of meteorology on air pollutant concentrations in urban area

    NASA Astrophysics Data System (ADS)

    Schäfer, Klaus; Blumenstock, Thomas; Bonn, Boris; Gerwig, Holger; Hase, Frank; Münkel, Christoph; Nothard, Rainer; von Schneidemesser, Erika

    2015-10-01

    Mixing layer height (MLH) is a key parameter to determine the influence of meteorological parameters upon air pollutants such as trace gas species and particulate concentrations near the surface. Meteorology, and MLH as a key parameter, affect the budget of emission source strengths, deposition, and accumulation. However, greater possibilities for the application of MLH data have been identified in recent years. Here, the results of measurements in Berlin in 2014 are shown and discussed. The concentrations of NO, NO2, O3, CO, PM1, PM2.5, PM10 and about 70 volatile organic compounds (anthropogenic and biogenic of origin) as well as particle size distributions and contributions of SOA and soot species to PM were measured at the urban background station of the Berlin air quality network (BLUME) in Nansenstr./Framstr., Berlin-Neukölln. A Vaisala ceilometer CL51, which is a commercial mini-lidar system, was applied at that site to detect the layers of the lower atmosphere in real time. Special software for these ceilometers with MATLAB provided routine retrievals of MLH from vertical profiles of laser backscatter data. Five portable Bruker EM27/SUN FTIR spectrometers were set up around Berlin to detect column averaged abundances of CO2 and CH4 by solar absorption spectrometry. Correlation analyses were used to show the coupling of temporal variations of trace gas compounds and PM with MLH. Significant influences of MLH upon NO, NO2, PM10, PM2.5, PM1 and toluene (marker for traffic emissions) concentrations as well as particle number concentrations in the size modes 70 - 100 nm, 100 - 200 nm and 200 - 500 nm on the basis of averaged diurnal courses were found. Further, MLH was taken as important auxiliary information about the development of the boundary layer during each day of observations, which was required for the proper estimation of CO2 and CH4 source strengths from Berlin on the basis of atmospheric column density measurements.

  11. Direct determination of crystallographic phases for diffraction data from lipid bilayers. II. Refinement of phospholipid structures.

    PubMed Central

    Dorset, D L

    1991-01-01

    Using a systematic approach for the acceptance of crystallographic phase assignment, based on the evaluation of triplet structure invariants, electron and x-ray diffraction data from phospholipid multilamellar arrays are analyzed by direct methods. After calculation of Fourier maps with a partial set of phased structure factor magnitudes, the structure is refined in real space by flattening of the hydrocarbon region of the bilayer and an optimal solution is sought either by the calculation of [delta rho 4] suggested by Luzzati, where rho is the structure density or by a test of density smoothness [magnitude of delta rho/ delta r magnitude of], where r positions are located along the normal to the lamellar surface. Reanalyses of previously determined structures sometimes lead to new conclusions (e.g., a possible similarity of the electron density profile for DL-DMPE and L-DMPE, and a clear indication of the fatty acid adduct in the mixed L-DPPC/palmitic acid bilayer). Because of presumed secondary scattering perturbations (primarily to the least intense reflections), the refinements of the electron diffraction intensities are less easily evaluated than those carried out with x-ray diffraction data. PMID:1777564

  12. Lipid nutritional value of legumes: Evaluation of different extraction methods and determination of fatty acid composition.

    PubMed

    Caprioli, Giovanni; Giusti, Federica; Ballini, Roberto; Sagratini, Gianni; Vila-Donat, Pilar; Vittori, Sauro; Fiorini, Dennis

    2016-02-01

    This study sought to contribute to the assessment of the nutritional properties of legumes by determining the fatty acid (FA) composition of 29 legume samples after the evaluation of nine extraction methods. The Folch method and liquid-solid extraction with hexane/isopropanol or with hexane/acetone were investigated, as was the effect of previous hydration of samples. Soxhlet extractions were also evaluated with different solvent mixtures. Results on FA composition using the hexane/isopropanol extraction method were the same in terms of FA composition of the Folch method, but the extraction yield was only around 20-40% of that of the Folch method preceded by hydration. Some types of legumes showed particularly interesting values for the ratio of polyunsaturated fatty acids (PUFAs) n-6/n-3, such as lentils, with the value of 4.0, and Azuki beans, at 3.2. In lentils, the PUFAs% ranged from 42.0% to 57.4%, while in Azuki beans it was 57.5%. PMID:26304436

  13. Age determination in manatees using growth-layer-group counts in bone

    USGS Publications Warehouse

    Marmontel, M.; O'Shea, T.J.; Kochman, H.I.; Humphrey, S.R.

    1996-01-01

    Growth layers were observed in histological preparations of bones of known-age, known minimum-age, and tetracycline-marked free-ranging and captive Florida manatees (Trichechus manatus latirostris), substantiating earlier preliminary findings of other studies. Detailed analysis of 17 new case histories showed that growth-layer group (GLG) counts in the periotic bone were consistent with known age, or time since tetracycline administration, but were less reliable in other bones. GLG counts were also made in periotic bones of 1,196 Florida manatees of unknown age found dead from 1974 through 1991. These counts were conducted in order to assess variability and to determine relationships among estimated age, size, sex, and degree of bone resorption. Resorption can interfere with accuracy of GLG counts. This effect does not occur until ages greater than about 15 yr and body lengths greater than 300 cm are attained. GLGs were also observed in periotic bones of Antillean manatees (Trichechus manatus manatus) but were not validated against known-age specimens. Use of GLG counts in the periotic bone is suitable for application to studies of population dynamics and other age-related aspects of manatee biology.

  14. Determining the Effective Density and Stabilizer Layer Thickness of Sterically Stabilized Nanoparticles

    PubMed Central

    2016-01-01

    A series of model sterically stabilized diblock copolymer nanoparticles has been designed to aid the development of analytical protocols in order to determine two key parameters: the effective particle density and the steric stabilizer layer thickness. The former parameter is essential for high resolution particle size analysis based on analytical (ultra)centrifugation techniques (e.g., disk centrifuge photosedimentometry, DCP), whereas the latter parameter is of fundamental importance in determining the effectiveness of steric stabilization as a colloid stability mechanism. The diblock copolymer nanoparticles were prepared via polymerization-induced self-assembly (PISA) using RAFT aqueous emulsion polymerization: this approach affords relatively narrow particle size distributions and enables the mean particle diameter and the stabilizer layer thickness to be adjusted independently via systematic variation of the mean degree of polymerization of the hydrophobic and hydrophilic blocks, respectively. The hydrophobic core-forming block was poly(2,2,2-trifluoroethyl methacrylate) [PTFEMA], which was selected for its relatively high density. The hydrophilic stabilizer block was poly(glycerol monomethacrylate) [PGMA], which is a well-known non-ionic polymer that remains water-soluble over a wide range of temperatures. Four series of PGMAx–PTFEMAy nanoparticles were prepared (x = 28, 43, 63, and 98, y = 100–1400) and characterized via transmission electron microscopy (TEM), dynamic light scattering (DLS), and small-angle X-ray scattering (SAXS). It was found that the degree of polymerization of both the PGMA stabilizer and core-forming PTFEMA had a strong influence on the mean particle diameter, which ranged from 20 to 250 nm. Furthermore, SAXS was used to determine radii of gyration of 1.46 to 2.69 nm for the solvated PGMA stabilizer blocks. Thus, the mean effective density of these sterically stabilized particles was calculated and determined to lie between 1.19 g

  15. Lidar determination of winds by aerosol inhomogeneities: motion velocity in the planetary boundary layer.

    PubMed

    Kolev, I; Parvanov, O; Kaprielov, B

    1988-06-15

    The paper presents results from lidar measurements of wind velocity in the planetary boundary layer using correlation data processing. Two lidars are used in our experiments: a ruby lidar operating along slant paths and a YAG:Nd lidar operating for near vertical sounding used by us for the first time. On the basis of our experience the optimal sizes of aerosol inhomogeneities (30-300 m), the duration of the experiments (2-10 min), and the repetition rate of laser shots (fractions of hertz to several hertz) are determined. The results are compared to independent data obtained from anemometer measurements, theodolite- and radar-tracked pilot balloons. The range of differences is ~1-2 m/s in speed and 10-15 degrees in direction. Preliminary results from the use of lidar data to remotely sound the wind speed for various atmospheric stratifications and synoptic situations are described as well. PMID:20531786

  16. A quantitative thin layer chromatography method for determination of theophylline in plasma.

    PubMed

    Mirfazaelian, A; Goudarzi, M; Tabatabaiefar, M; Mahmoudian, M

    2002-01-01

    A simple assay method for theophylline in plasma using thin layer chromatography (TLC) was developed. The method involves extraction of the drug and internal standard (acetaminophen) by chloroform-isopropanol (75:25) followed by separation on TLC silica plates using a mixture of acetic acid, isopropanol, toluene (1: 12: 6), as the eluting solvent. Both peak height ratios and peak are ratios showed high correlation coefficient (r>0.98, p<0.001). However we used peak heights for the determinations. Within-day and between-day coefficients of variation were less than 4.4% and 7.8% respectively. The assay proved inexpensive, accurate and reproducible with a limit of detection of 100 ng/ml that makes it suitable for bioavailability studies.

  17. Determination of the thickness and orientation of few-layer tungsten ditelluride using polarized Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Kim, Minjung; Han, Songhee; Kim, Jung Hwa; Lee, Jae-Ung; Lee, Zonghoon; Cheong, Hyeonsik

    2016-09-01

    Orthorhombic tungsten ditelluride (WTe2), with a distorted 1T structure, exhibits a large magnetoresistance that depends on the orientation, and its electrical characteristics changes from semimetallic to insulating as the thickness decreases. Through polarized Raman spectroscopy in combination with transmission electron diffraction, we establish a reliable method to determine the thickness and crystallographic orientation of few-layer WTe2. The Raman spectrum shows a pronounced dependence on the polarization of the excitation laser. We found that the separation between two Raman peaks at ∼90 cm‑1 and at 80–86 cm‑1, depending on thickness, is a reliable fingerprint for determination of the thickness. For determination of the crystallographic orientation, the polarization dependence of the A 1 modes, measured with the 632.8 nm excitation, turns out to be the most reliable. We also discovered that the polarization behaviors of some of the Raman peaks depend on the excitation wavelength as well as thickness, indicating a close interplay between the band structure and anisotropic Raman scattering cross section.

  18. Determination of the thickness and orientation of few-layer tungsten ditelluride using polarized Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Kim, Minjung; Han, Songhee; Kim, Jung Hwa; Lee, Jae-Ung; Lee, Zonghoon; Cheong, Hyeonsik

    2016-09-01

    Orthorhombic tungsten ditelluride (WTe2), with a distorted 1T structure, exhibits a large magnetoresistance that depends on the orientation, and its electrical characteristics changes from semimetallic to insulating as the thickness decreases. Through polarized Raman spectroscopy in combination with transmission electron diffraction, we establish a reliable method to determine the thickness and crystallographic orientation of few-layer WTe2. The Raman spectrum shows a pronounced dependence on the polarization of the excitation laser. We found that the separation between two Raman peaks at ˜90 cm-1 and at 80-86 cm-1, depending on thickness, is a reliable fingerprint for determination of the thickness. For determination of the crystallographic orientation, the polarization dependence of the A 1 modes, measured with the 632.8 nm excitation, turns out to be the most reliable. We also discovered that the polarization behaviors of some of the Raman peaks depend on the excitation wavelength as well as thickness, indicating a close interplay between the band structure and anisotropic Raman scattering cross section.

  19. Determination of free fatty acids in pharmaceutical lipids by ¹H NMR and comparison with the classical acid value.

    PubMed

    Skiera, Christina; Steliopoulos, Panagiotis; Kuballa, Thomas; Diehl, Bernd; Holzgrabe, Ulrike

    2014-05-01

    Indices like acid value, peroxide value, and saponification value play an important role in quality control and identification of lipids. Requirements on these parameters are given by the monographs of the European pharmacopeia. (1)H NMR spectroscopy provides a fast and simple alternative to these classical approaches. In the present work a new (1)H NMR approach to determine the acid value is described. The method was validated using a statistical approach based on a variance components model. The performance under repeatability and in-house reproducibility conditions was assessed. We applied this (1)H NMR assay to a wide range of different fatty oils. A total of 305 oil and fat samples were examined by both the classical and the NMR method. Except for hard fat, the data obtained by the two methods were in good agreement. The (1)H NMR method was adapted to analyse waxes and oleyloleat. Furthermore, the effect of solvent and in the case of castor oil the effect of the oil matrix on line broadening and chemical shift of the carboxyl group signal are discussed.

  20. Reconstitution baking tests with defatted wheat flour are suitable for determining the functional effects of lipase-treated wheat lipids.

    PubMed

    Schaffarczyk, Monika; Østdal, Henrik; Matheis, Olivia; Jekle, Mario; Koehler, Peter

    2016-06-01

    A microscale reconstitution baking test, using wheat flour defatted with 2-propanol at 20 °C, was established to determine the functional effects of lipids isolated from lipase-treated wheat dough. Proper selection of solvent and extraction temperature was of major importance to maintain the functionality of defatted flour. Dough and gluten from flour defatted with water-saturated 1-butanol (WSB; extracted at 20 °C) and 2-propanol (extracted at 75 °C) had inferior extensibility and loaf volume compared to control flour extracted with 2-propanol at 20 °C. Quantitation of gluten proteins showed that defatting with WSB (20 °C) or 2-propanol (75 °C) decreased the gliadin and increased the glutenin content. Possible reasons were thiol-disulfide interchange reactions, caused either by heat (2-propanol, 75 °C) or by the solvent WSB, which affected gluten proteins. Confocal laser scanning microscopy showed that regular, interconnected gluten structures were only present in dough from flour defatted with 2-propanol at 20 °C. PMID:26830576

  1. Reconstitution baking tests with defatted wheat flour are suitable for determining the functional effects of lipase-treated wheat lipids.

    PubMed

    Schaffarczyk, Monika; Østdal, Henrik; Matheis, Olivia; Jekle, Mario; Koehler, Peter

    2016-06-01

    A microscale reconstitution baking test, using wheat flour defatted with 2-propanol at 20 °C, was established to determine the functional effects of lipids isolated from lipase-treated wheat dough. Proper selection of solvent and extraction temperature was of major importance to maintain the functionality of defatted flour. Dough and gluten from flour defatted with water-saturated 1-butanol (WSB; extracted at 20 °C) and 2-propanol (extracted at 75 °C) had inferior extensibility and loaf volume compared to control flour extracted with 2-propanol at 20 °C. Quantitation of gluten proteins showed that defatting with WSB (20 °C) or 2-propanol (75 °C) decreased the gliadin and increased the glutenin content. Possible reasons were thiol-disulfide interchange reactions, caused either by heat (2-propanol, 75 °C) or by the solvent WSB, which affected gluten proteins. Confocal laser scanning microscopy showed that regular, interconnected gluten structures were only present in dough from flour defatted with 2-propanol at 20 °C.

  2. Determination of free fatty acids in pharmaceutical lipids by ¹H NMR and comparison with the classical acid value.

    PubMed

    Skiera, Christina; Steliopoulos, Panagiotis; Kuballa, Thomas; Diehl, Bernd; Holzgrabe, Ulrike

    2014-05-01

    Indices like acid value, peroxide value, and saponification value play an important role in quality control and identification of lipids. Requirements on these parameters are given by the monographs of the European pharmacopeia. (1)H NMR spectroscopy provides a fast and simple alternative to these classical approaches. In the present work a new (1)H NMR approach to determine the acid value is described. The method was validated using a statistical approach based on a variance components model. The performance under repeatability and in-house reproducibility conditions was assessed. We applied this (1)H NMR assay to a wide range of different fatty oils. A total of 305 oil and fat samples were examined by both the classical and the NMR method. Except for hard fat, the data obtained by the two methods were in good agreement. The (1)H NMR method was adapted to analyse waxes and oleyloleat. Furthermore, the effect of solvent and in the case of castor oil the effect of the oil matrix on line broadening and chemical shift of the carboxyl group signal are discussed. PMID:23664852

  3. Epidermal surface lipids

    PubMed Central

    2009-01-01

    A layer of lipids, which are of both sebaceous and keratinocyte origin, covers the surface of the skin. The apparent composition of surface lipids varies depending on the selected method of sampling. Lipids produced by the epidermal cells are an insignificant fraction of the total extractable surface lipid on areas rich in sebaceous glands. Due to the holocrine activity of the sebaceous gland, its product of secretion (sebum) is eventually released to the surface of the skin and coats the fur as well. Lipids of epidermal origin fill the spaces between the cells, like mortar or cement. The sebaceous lipids are primarily non polar lipids as triglycerides, wax esters and squalene, while epidermal lipids are a mixture of ceramides, free fatty acids and cholesterol. The composition of the sebaceous lipids is unique and intriguing and elevated sebum excretion is a major factor involved in the pathophysiology of acne. Recent studies have elucidated the roles that epidermal surface lipids have on normal skin functions and acne. PMID:20224687

  4. Rapid determination of chlorinated pesticides in fish by freezing-lipid filtration, solid-phase extraction and gas chromatography-mass spectrometry.

    PubMed

    Hong, Jongki; Kima, Hye-Young; Kim, Do-Gyun; Seo, Jungju; Kimb, Kang-Jin

    2004-06-01

    An analytical method has been developed for measuring 24 chlorinated pesticides in fish tissue samples. Extraction of chlorinated pesticides was carried out by ultrasonication using an acetone-n-hexane (5:2, v/v) mixture. Most of the lipids in the extract were eliminated by freezing-lipid filtration, prior to solid-phase extraction (SPE) cleanup. During freezing-lipid filtration, about 90% of the lipids extracted from the fish samples were easily removed without any significant losses of chlorinated pesticides. For purification, SPE using Florisil was shown to be more effective than silica. Quantification was performed using gas chromatography-mass spectrometry in the selected ion monitoring mode. Spiking experiments were carried out to determine the recovery, precision, and limits of detection (LODs) of the method. The overall recovery was above 80% in the spiked fish tissue sample at 100 ng/g level. The detection limits for chlorinated pesticides were ranged from 0.5 to 5 ppb, except for endosulfan I and II which was 20 ppb. The newly developed method is demonstrated to give efficient recoveries and LODs for detecting chlorinated pesticides spiked into fish tissue with high lipid content.

  5. Lipid compositions in Escherichia coli and Bacillus subtilis during growth as determined by MALDI-TOF and TOF/TOF mass spectrometry

    NASA Astrophysics Data System (ADS)

    Gidden, Jennifer; Denson, Jackie; Liyanage, Rohana; Ivey, D. Mack; Lay, Jackson O., Jr.

    2009-06-01

    Lipids in Escherichia coli and Bacillus subtilis were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and TOF/TOF tandem mass spectrometry. Lipids were extracted from bacterial cells using an equal volume mixture of dichloromethane, ethanol, and water, which formed a biphasic system with the lipids in the organic layer. The resulting mass spectra of the extracts from both bacteria showed a series of peaks corresponding to sodiated phospholipids - primarily phosphatidylethanolamines (PEs) and phosphatidylglycerols (PGs). The relative amounts of the phospholipids and the fatty acid compositions inferred from the spectra were in good agreement with previously reported values from GC/MS and thin-layer chromatography studies. E. coli and B. subtilis were easily differentiated by dissimilarities in the composition and relative amounts of the phospholipids present as well as by the presence of lysyl-phosphatidylglycerol and diglycosyl diglycerides solely in the B. subtilis mass spectra. Changes in lipid content in the bacteria during their growth phases were also monitored. In E. coli, the spectra indicated an increase in the amount of the unique Ccy-17 fatty acid (in which the fatty acid chain contains a cyclopropane ring) formed during exponential growth. During stationary growth, the spectra indicated an increase in the amount of saturated fatty acids. In B. subtilis, the phospholipid composition remained relatively unchanged during exponential growth, but the amount of PG slightly decreased while the amount of PE slightly increased during stationary growth. No significant changes were observed for the lysyl-phosphatidylglycerols or glycolipids during the exponential or stationary growth phases.

  6. Simultaneous determination of cyclodol and diprazin by thin layer chromatography and high performance liquid chromatography.

    PubMed

    Makharadze, R; Adeishvili, L; Chelidze, T; Imnadze, N; Nizharadze, N

    2009-11-01

    Ciklodol (trihexyphenidil)--the central and peripheral m-cholinoblocker is currently used with other antipsychotic drugs such as phenotiazines and tricycle antidepressants. For the purpose of simultaneous determination of ciklodol and diprazine, were selected two methods of analysis: Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). During development of TLC method was studied the 10 visualizing system and 24 mobile systems. For individual or simultaneous determination of ciklodol and diprazine were recommended the following solvents' systems: 1. Toluene-acetone-ethanole-25%NH(4)OH (45:45: 7.5:2.5), 2. Hexane-ethyl acetate (15:5), 3. Chloroform-heptene-25%NH(4)OH (16:3:3), 4. Ethylacetate-hexane (10:10), 5. Acetonitrile-metanol (10:10) and 6.Heptene-chloroform-ethanol-25% NH(4)OH (5:10:3:1). As visualizing systems were chosen: Iodine vapors, blacklight (UV254) and reagent of FNP. Reagent of FNP gives colored spot just with diprazine and it is also could be used for separation of both objects in simultaneous analysis. Developed HPLC method of simultaneous determination of ciklodol and diprazine: like mobile phase is recommended: Acetonitril- 0.05M KH(2)PO4 (55:45) (v/v) +H(3)PO(4) (pH3.5), column EC250 x 4.6mm, with solid phase Nucleosil, flow rate 1ml/min, sample volume 40 microl. In given conditions, the retention time of ciklodol is 6.005min and diprazine 7.227min. Developed method of simultaneous determination and separation of ciklodol and diprazine in respective mixtures could be successfully applied as in the pharmaceutical, as well in the chemical-toxicological laboratories.

  7. Surface layer determination for the Si spheres of the Avogadro project

    NASA Astrophysics Data System (ADS)

    Busch, I.; Azuma, Y.; Bettin, H.; Cibik, L.; Fuchs, P.; Fujii, K.; Krumrey, M.; Kuetgens, U.; Kuramoto, N.; Mizushima, S.

    2011-04-01

    For the accurate determination of the Avogadro constant, two 28Si spheres were produced, whose macroscopic density, in addition to other values, must be determined. To make a contribution to the new definition of the kilogram, a relative standard uncertainty of less than 2 × 10-8 has to be achieved. Each silicon surface is covered by a surface layer (SL). Consequently, correction parameters for the SL are determined to be applied to the mass and volume determination of the enriched spheres. With the use of a large set of surface analysing techniques, the structure of the SL is investigated. An unexpected metallic contamination existing on the sphere surface enlarges the uncertainty contribution of the correction parameters above the originally targeted value of 1 × 10-8. In the framework of this investigation this new obstacle is resolved in two ways. A new combination of analytical methods is applied to measure the SL mass mSL and the thickness dSL, including this new contamination, with an uncertainty of u(mSL) = 14.5 µg and 14.4 µg, respectively, and u(dSL) = 0.33 nm and 0.32 nm for the 28Si spheres AVO28-S5 and AVO28-S8, respectively. In the second part of the work, the chemical composition of these metallic contaminations is found to be Cu, Ni and Zn silicide compounds. For the removal of this contamination, a special procedure is developed, tested and applied to the spheres to produce the originally expected surface structure on the spheres. After the application of this new procedure the use of x-ray reflectometry directly at the spheres will be possible. It is expected to reduce the uncertainty contribution due to the SL down to 1 × 10-8.

  8. "Density equilibrium" method for the quantitative and rapid in situ determination of lipid, hydrocarbon, or biopolymer content in microorganisms.

    PubMed

    Eroglu, Ela; Melis, Anastasios

    2009-04-01

    The work provides a simple method, based on a direct density equilibrium measurement, for the rapid in situ estimation of total lipid, hydrocarbon or biopolymer content in a variety of prokaryotic and eukaryotic samples. The method can be readily applied to live microalgae and photosynthetic bacteria, single-celled or colonial microorganisms, as well as cellular fractions and isolated subcellular compartments or components. In this approach, the absolute lipid, hydrocarbon, or biopolymer content of the cells can be readily calculated. This method is especially useful for tracking the oil or polymer content of strains of microalgae and other microorganisms, whose lipid, hydrocarbon or biopolymer content may change with cultivation conditions and/or time, as the case would be in microorganism lipid-induction industrial processes. The method is also useful for the direct in situ measurement of storage polymer accumulation in live cells, such as starch in microalgae and polyhydroxybutyrate, or other polyhydroxyalkanoates, in photosynthetic and non-photosynthetic bacteria. PMID:19031427

  9. The association of thromboxane A2 receptor with lipid rafts is a determinant for platelet functional responses.

    PubMed

    Moscardó, A; Vallés, J; Latorre, A; Santos, M T

    2014-08-25

    We have investigated the presence of thromboxane A2 (TXA2) receptor associated with lipid rafts in human platelets and the regulation of platelet function in response to TXA2 receptor agonists when lipid rafts are disrupted by cholesterol extraction. Platelet aggregation with TXA2 analogs U46619 and IBOP was almost blunted in cholesterol-depleted platelets, as well as αIIbβ3 integrin activation and P-selectin exposure. Raft disruption also inhibited TXA2-induced cytosolic calcium increase and nucleotide release, ruling out an implication of P2Y12 receptor. An important proportion of TXA2 receptor (40%) was colocalized at lipid rafts. The presence of the TXA2 receptor associated with lipid rafts in platelets is important for functional platelet responses to TXA2.

  10. Quantitative Determination of Photosynthetic Pigments in Green Beans Using Thin-Layer Chromatography and a Flatbed Scanner as Densitometer

    ERIC Educational Resources Information Center

    Valverde, Juan; This, Herve; Vignolle, Marc

    2007-01-01

    A simple method for the quantitative determination of photosynthetic pigments extracted from green beans using thin-layer chromatography is proposed. Various extraction methods are compared, and it is shown how a simple flatbed scanner and free software for image processing can give a quantitative determination of pigments. (Contains 5 figures.)

  11. Lipid body formation plays a central role in cell fate determination during developmental differentiation of Myxococcus xanthus.

    PubMed

    Hoiczyk, Egbert; Ring, Michael W; McHugh, Colleen A; Schwär, Gertrud; Bode, Edna; Krug, Daniel; Altmeyer, Matthias O; Lu, Jeff Zhiqiang; Bode, Helge B

    2009-10-01

    Cell differentiation is widespread during the development of multicellular organisms, but rarely observed in prokaryotes. One example of prokaryotic differentiation is the gram-negative bacterium Myxococcus xanthus. In response to starvation, this gliding bacterium initiates a complex developmental programme that results in the formation of spore-filled fruiting bodies. How the cells metabolically support the necessary complex cellular differentiation from rod-shaped vegetative cells into spherical spores is unknown. Here, we present evidence that intracellular lipid bodies provide the necessary metabolic fuel for the development of spores. Formed at the onset of starvation, these lipid bodies gradually disappear until they are completely used up by the time the cells have become mature spores. Moreover, it appears that lipid body formation in M. xanthus is an important initial step indicating cell fate during differentiation. Upon starvation, two subpopulations of cells occur: cells that form lipid bodies invariably develop into spores, while cells that do not form lipid bodies end up becoming peripheral rods, which are cells that lack signs of morphological differentiation and stay in a vegetative-like state. These data indicate that lipid bodies not only fuel cellular differentiation but that their formation represents the first known morphological sign indicating cell fate during differentiation. PMID:19788540

  12. Lipid body formation plays a central role in cell fate determination during developmental differentiation of Myxococcus xanthus

    PubMed Central

    Ring, Michael W.; McHugh, Colleen A.; Schwär, Gertrud; Bode, Edna; Krug, Daniel; Altmeyer, Matthias O.; Lu, Jeff Zhiqiang

    2010-01-01

    Summary Cell differentiation is widespread during the development of multicellular organisms, but rarely observed in prokaryotes. One example of prokaryotic differentiation is the Gramnegative bacterium Myxococcus xanthus. In response to starvation, this gliding bacterium initiates a complex developmental program that results in the formation of spore-filled fruiting bodies. How the cells metabolically support the necessary complex cellular differentiation from rod-shaped vegetative cells into spherical spores is unknown. Here, we present evidence that intra-cellular lipid bodies provide the necessary metabolic fuel for the development of spores. Formed at the onset of starvation, these lipid bodies gradually disappear until they are completely used up by the time the cells have become mature spores. Moreover, it appears that lipid body formation in M. xanthus is an important initial step indicating cell fate during differentiation. Upon starvation, two subpopulations of cells occur: cells that form lipid bodies invariably develop into spores, while cells that do not form lipid bodies end up becoming peripheral rods, which are cells that lack signs of morphological differentiation and stay in a vegetative-like state. These data indicate that lipid bodies not only fuel cellular differentiation but that their formation represents the first known morphological sign indicating cell fate during differentiation. PMID:19788540

  13. Determination of effective mechanical properties of a double-layer beam by means of a nano-electromechanical transducer

    SciTech Connect

    Hocke, Fredrik; Pernpeintner, Matthias; Gross, Rudolf; Zhou, Xiaoqing; Kippenberg, Tobias J.; Schliesser, Albert; Huebl, Hans

    2014-09-29

    We investigate the mechanical properties of a doubly clamped, double-layer nanobeam embedded into an electromechanical system. The nanobeam consists of a highly pre-stressed silicon nitride and a superconducting niobium layer. By measuring the mechanical displacement spectral density both in the linear and the nonlinear Duffing regime, we determine the pre-stress and the effective Young's modulus of the nanobeam. An analytical double-layer model quantitatively corroborates the measured values. This suggests that this model can be used to design mechanical multilayer systems for electro- and optomechanical devices, including materials controllable by external parameters such as piezoelectric, magnetostrictive, or in more general multiferroic materials.

  14. Determination of effective mechanical properties of a double-layer beam by means of a nano-electromechanical transducer

    NASA Astrophysics Data System (ADS)

    Hocke, Fredrik; Pernpeintner, Matthias; Zhou, Xiaoqing; Schliesser, Albert; Kippenberg, Tobias J.; Huebl, Hans; Gross, Rudolf

    2014-09-01

    We investigate the mechanical properties of a doubly clamped, double-layer nanobeam embedded into an electromechanical system. The nanobeam consists of a highly pre-stressed silicon nitride and a superconducting niobium layer. By measuring the mechanical displacement spectral density both in the linear and the nonlinear Duffing regime, we determine the pre-stress and the effective Young's modulus of the nanobeam. An analytical double-layer model quantitatively corroborates the measured values. This suggests that this model can be used to design mechanical multilayer systems for electro- and optomechanical devices, including materials controllable by external parameters such as piezoelectric, magnetostrictive, or in more general multiferroic materials.

  15. Birth order dependent growth cone segregation determines synaptic layer identity in the Drosophila visual system

    PubMed Central

    Kulkarni, Abhishek; Ertekin, Deniz; Lee, Chi-Hon; Hummel, Thomas

    2016-01-01

    The precise recognition of appropriate synaptic partner neurons is a critical step during neural circuit assembly. However, little is known about the developmental context in which recognition specificity is important to establish synaptic contacts. We show that in the Drosophila visual system, sequential segregation of photoreceptor afferents, reflecting their birth order, lead to differential positioning of their growth cones in the early target region. By combining loss- and gain-of-function analyses we demonstrate that relative differences in the expression of the transcription factor Sequoia regulate R cell growth cone segregation. This initial growth cone positioning is consolidated via cell-adhesion molecule Capricious in R8 axons. Further, we show that the initial growth cone positioning determines synaptic layer selection through proximity-based axon-target interactions. Taken together, we demonstrate that birth order dependent pre-patterning of afferent growth cones is an essential pre-requisite for the identification of synaptic partner neurons during visual map formation in Drosophila. DOI: http://dx.doi.org/10.7554/eLife.13715.001 PMID:26987017

  16. Determination of contact angles on microporous particles using the thin-layer wicking technique.

    PubMed

    Cui, Zheng-Gang; Binks, Bernard P; Clint, John H

    2005-08-30

    The properties of particle-stabilized emulsions, especially with regard to phase inversion, are very dependent on the contact angle that the particles experience at the oil-water interface. For the very small particles used for such emulsions (often a few tens of nm), it is impossible to measure this contact angle directly. Its value could be calculated if it were possible to determine the components of the solid surface free energy. To establish a method suitable for such particles, we have investigated the imbibition of five probe liquids into a porous bed of silica (commercial TLC plates) using the thin-layer wicking technique. For all liquids, the difference between wicking rate for bare plates and for those pre-contacted with the vapors is large but it is not due to an advancing angle effect on bare plates. Our analysis shows that it is due to the diversion of flowing liquid into blind pores which are already filled in the pre-contacted case. Thus a new model is proposed describing wicking in a porous medium with very small blind pores by introducing a parameter into the Washburn equation that corrects for this capillary condensation effect. The parameter needed is determined independently using gravimetric adsorption measurements. When this modified Washburn equation is used, the difference between advancing and receding contact angle is actually quite small. When the averages are used as the Young's contact angles, values for the surface energy components of silica are obtained that are completely consistent between the five liquids and have magnitudes expected for this type of silica surface.

  17. Thin-layer chromatography of aluminium: quantitative densitometric determination of Fe2+, Ni2+, Cu2+, and Si4+.

    PubMed

    Mohamed Najar, P A; Chouhan, R N; Jeurkar, J U; Dolas, S D; Ramana Rao, K V

    2007-01-01

    Thin-layer chromatography in combination with scanning densitometry is used as a tool for the quantitative determination of some impurity and additive elements in aluminium. Microgram levels of iron, silicon, copper, nickel, titanium, magnesium, manganese, and zinc present in a high concentration aluminium matrix is detected, and selective separations of some of these elements are achieved on silica gel H layers developed with a mobile phase containing aqueous sodium chloride solution. The quantitative determination of iron, silicon, nickel, and copper are obtained from the densitometric evaluation of chromatograms and are compared with the respective optical emission spectral analytical data. PMID:17555634

  18. Conformation transformation determined by different self-assembled phases in a DNA complex with cationic polyhedral oligomeric silsesquioxane lipid.

    PubMed

    Cui, Li; Chen, Daoyong; Zhu, Lei

    2008-05-01

    In this work, a novel cube-shaped cationic lipid based on the imidazolium salt of polyhedral oligomeric silsesquioxane (POSS) was complexed with double-stranded DNA. Because of the negative spontaneous curvature of the cationic POSS imidazolium lipid, an inverted hexagonal phase resulted above the melting point of POSS crystals. Depending on the competition between the crystallization of POSS molecules and the negative spontaneous curvature of cationic POSS imidazolium lipids, different self-assembled phase morphologies were obtained. A lamellar phase was obtained when the POSS crystallization was relatively slow. When the POSS crystallization was fast, an inverted hexagonal phase was obtained with POSS lamellar crystals grown in the interstitials of DNA cylinders. On the basis of a circular dichroism study, double-stranded DNA adopted the B-form helical conformation in the inverted hexagonal phase, whereas the helical conformation was largely destroyed in the lamellar phase.

  19. Oral mucosal lipids are antibacterial against Porphyromonas gingivalis, induce ultrastructural damage, and alter bacterial lipid and protein compositions

    PubMed Central

    Fischer, Carol L; Walters, Katherine S; Drake, David R; Dawson, Deborah V; Blanchette, Derek R; Brogden, Kim A; Wertz, Philip W

    2013-01-01

    Oral mucosal and salivary lipids exhibit potent antimicrobial activity for a variety of Gram-positive and Gram-negative bacteria; however, little is known about their spectrum of antimicrobial activity or mechanisms of action against oral bacteria. In this study, we examine the activity of two fatty acids and three sphingoid bases against Porphyromonas gingivalis, an important colonizer of the oral cavity implicated in periodontitis. Minimal inhibitory concentrations, minimal bactericidal concentrations, and kill kinetics revealed variable, but potent, activity of oral mucosal and salivary lipids against P. gingivalis, indicating that lipid structure may be an important determinant in lipid mechanisms of activity against bacteria, although specific components of bacterial membranes are also likely important. Electron micrographs showed ultrastructural damage induced by sapienic acid and phytosphingosine and confirmed disruption of the bacterial plasma membrane. This information, coupled with the association of treatment lipids with P. gingivalis lipids revealed via thin layer chromatography, suggests that the plasma membrane is a likely target of lipid antibacterial activity. Utilizing a combination of two-dimensional in-gel electrophoresis and Western blot followed by mass spectroscopy and N-terminus degradation sequencing we also show that treatment with sapienic acid induces upregulation of a set of proteins comprising a unique P. gingivalis stress response, including proteins important in fatty acid biosynthesis, metabolism and energy production, protein processing, cell adhesion and virulence. Prophylactic or therapeutic lipid treatments may be beneficial for intervention of infection by supplementing the natural immune function of endogenous lipids on mucosal surfaces. PMID:23867843

  20. Evaluating quality and its determinants in lipid control for secondary prevention of heart disease and stroke in primary care: a study in an inner London Borough

    PubMed Central

    Dodhia, Hiten; Kun, Liu; Logan Ellis, Hugh; Crompton, James; Wierzbicki, Anthony S; Williams, Helen; Hodgkinson, Anna; Balazs, John

    2015-01-01

    Objectives To assess quality of management and determinants in lipid control for secondary prevention of cardiovascular disease (CVD) using multilevel regression models. Design Cross-sectional study. Setting Inner London borough, with a primary care registered population of 378 000 (2013). Participants 48/49 participating general practices with 7869 patients on heart disease/stroke registers were included. Outcome measures (1) Recording of current total cholesterol levels and lipid control according to national evidence-based standards. (2) Assessment of quality by age, sex, ethnicity, deprivation, presence of other risks or comorbidity in meeting both lipid measurement and control standards. Results Some process standards were not met. Patients with a current cholesterol measurement >5 mmol/L were less likely to have a current statin prescription (adjusted OR=3.10; 95% CI 2.70 to 3.56). They were more likely to have clustering of other CVD risk factors. Women were significantly more likely to have raised cholesterol after adjustment for other factors (adjusted OR=1.74; 95% CI 1.53 to 1.98). Conclusions In this study, the key factor that explained poor lipid control in people with CVD was having no current prescription record of a statin. Women were more likely to have poorly controlled cholesterol (independent of comorbid risk factors and after adjusting for age, ethnicity, deprivation index and practice-level variation). Women with CVD should be offered statin prescription and may require higher statin dosage for improved control. PMID:26656014

  1. Optimization of the C11-BODIPY(581/591) dye for the determination of lipid oxidation in Chlamydomonas reinhardtii by flow cytometry.

    PubMed

    Cheloni, Giulia; Slaveykova, Vera I

    2013-10-01

    Lipid oxidation is a recognized end point for the study of oxidative stress and is an important parameter to describe the mode of micropollutant action on aquatic microorganisms. Therefore, the development of quick and reliable methodologies probing the oxidative stress and damage in living cells is highly sought. In the present proof-of-concept work, we examined the potential of the fluorescent dye C11-BODIPY(591/581) to probe lipid oxidation in the green microalga Chlamydomonas reinhardtii. C11-BODIPY(591/581) staining was combined with flow cytometry measurements to obtain multiparameter information on cellular features and oxidative stress damage within single cells. First, staining conditions were optimized by exploring the capability of the dye to stain algal cells under increasing cell and dye concentrations and different staining procedures. Then lipid oxidation in algae induced by short- and long-term exposures to the three metallic micropollutants, copper, mercury, and nanoparticulate copper oxide, and the two organic contaminants, diethyldithiocarbamate (DDC) and diuron was determined. In this work we pointed out C11-BODIPY(591/581) applicability in a wide range of exposure conditions, including studies of oxidation as a function of time and that it is suitable for in vivo measurements of lipid oxidation due to its high permeation and stability in cells and its low interference with algal autofluorescence. © 2013 International Society for Advancement of Cytometry. PMID:23943236

  2. In Vitro Amphotericin B Susceptibility of Malassezia pachydermatis Determined by the CLSI Broth Microdilution Method and Etest Using Lipid-Enriched Media

    PubMed Central

    Álvarez-Pérez, Sergio; Peláez, Teresa; Cutuli, Maite; García, Marta E.

    2014-01-01

    We determined the in vitro amphotericin B susceptibility of 60 Malassezia pachydermatis isolates by the CLSI broth microdilution method and the Etest using lipid-enriched media. All isolates were susceptible at MICs of ≤1 μg/ml, confirming the high activity of amphotericin B against this yeast species. Overall, the essential agreement between the tested methods was high (80% and 96.7% after 48 h and 72 h, respectively), and all discrepancies were regarded as nonsubstantial. PMID:24752258

  3. Alteration of interleaflet coupling due to compounds displaying rapid translocation in lipid membranes

    PubMed Central

    Reigada, Ramon

    2016-01-01

    The spatial coincidence of lipid domains at both layers of the cell membrane is expected to play an important role in many cellular functions. Competition between the surface interleaflet tension and a line hydrophobic mismatch penalty are conjectured to determine the transversal behavior of laterally heterogeneous lipid membranes. Here, by a combination of molecular dynamics simulations, a continuum field theory and kinetic equations, I demonstrate that the presence of small, rapidly translocating molecules residing in the lipid bilayer may alter its transversal behavior by favoring the spatial coincidence of similar lipid phases. PMID:27596355

  4. Alteration of interleaflet coupling due to compounds displaying rapid translocation in lipid membranes

    NASA Astrophysics Data System (ADS)

    Reigada, Ramon

    2016-09-01

    The spatial coincidence of lipid domains at both layers of the cell membrane is expected to play an important role in many cellular functions. Competition between the surface interleaflet tension and a line hydrophobic mismatch penalty are conjectured to determine the transversal behavior of laterally heterogeneous lipid membranes. Here, by a combination of molecular dynamics simulations, a continuum field theory and kinetic equations, I demonstrate that the presence of small, rapidly translocating molecules residing in the lipid bilayer may alter its transversal behavior by favoring the spatial coincidence of similar lipid phases.

  5. Alteration of interleaflet coupling due to compounds displaying rapid translocation in lipid membranes.

    PubMed

    Reigada, Ramon

    2016-01-01

    The spatial coincidence of lipid domains at both layers of the cell membrane is expected to play an important role in many cellular functions. Competition between the surface interleaflet tension and a line hydrophobic mismatch penalty are conjectured to determine the transversal behavior of laterally heterogeneous lipid membranes. Here, by a combination of molecular dynamics simulations, a continuum field theory and kinetic equations, I demonstrate that the presence of small, rapidly translocating molecules residing in the lipid bilayer may alter its transversal behavior by favoring the spatial coincidence of similar lipid phases. PMID:27596355

  6. An ultrasonic theoretical and experimental approach to determine thickness and wave speed in layered media.

    PubMed

    de Sousa, Ana Valéria Greco; Pereira, Wagner Coelho de Albuquerque; Machado, João Carlos

    2007-02-01

    This work presents an ultrasonic method to characterize the layers of a stratified medium, using independent measurements of wave speed and thickness of each layer. The model, based on geometrical acoustics, includes refraction. Two transducers are used: one active (3.4 MHz) and a hydrophone as a receptor, which is moved laterally through 15 positions. The distance between the transducers and the delay between the echoes, from the interfaces separating the layers, received by them are used to estimate the speed and thickness. Three types of layered phantoms were used: Ph1 made with alcohol/acrylic, Ph2 made with polyvinyl chloride/water/acrylic, and Ph3 made with acrylic/water/polyvinyl chloride. The experimental results for speed of sound and layer thickness presented an experimental mean relative error, for thickness and wave speed, lower than 7.0% and 6.6%, respectively. PMID:17328335

  7. Determination of solid state characteristics of spray-congealed Ibuprofen solid lipid microparticles and their impact on sustaining drug release.

    PubMed

    Wong, Priscilla Chui Hong; Heng, Paul Wan Sia; Chan, Lai Wah

    2015-05-01

    This study was used to find solid state characteristics of ibuprofen loaded spray-congealed solid lipid microparticles (SLMs) by employing simple lipids as matrices, with or without polymeric additives, and the impact of solid drug-matrix miscibility on sustaining drug release. Solid miscibility of ibuprofen with two lipids, cetyl alcohol (CA) and stearic acid (SA), were investigated using differential scanning calorimetry (DSC). SLMs containing 20% w/w ibuprofen with or without polymeric additives, PVP/VA and EC, were produced by spray congealing, and the resultant microparticles were subjected to visual examination by scanning electron microscopy (SEM), thermal analysis using DSC, and hot-stage microscopy. Intermolecular interactions between lipids and drug as well as additives were investigated by Fourier-transformed infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR). X-ray diffractometry (XRD) was utilized to study polymorphic changes of drug and matrix over the course of a year. Ibuprofen was found to depress the melting points of CA and SA in a colligative manner, reaching maximum solubility at 10% w/w and 30% w/w for CA and SA, respectively. Drug encapsulation efficiencies and yields of spray-congealed SLMs containing 20% w/w ibuprofen were consistently high for both lipid matrices. CA and SA were found to adopt their stable γ- and β-polymorphs, respectively, immediately after spray congealing. The spray congealing process resulted in ibuprofen adopting an amorphous or poorly crystalline state, with no further changes over the course of a year. SEM, DSC, and hot stage microscope studies on the SLMs confirmed the formation of a solid dispersion between ibuprofen and CA and a solid solution between ibuprofen and SA. SA was found to sustain the release of ibuprofen significantly better than CA. PVP/VA and EC showed some interactions with CA, which led to an expansion of unit cell dimensions of CA upon spray congealing, whereas they

  8. INAA determination of major and trace elements in loess, paleosol and precipitation layers in a pleistocene Loess Section, China

    USGS Publications Warehouse

    Tian, J.; Chou, C.-L.; Ehmann, W.D.

    1987-01-01

    Instrumental neutron activation analysis was used for the determination of 31 major and trace elements in 32 samples from the Xinji Loess Section, Shaanxi Province, China. Interferences, including those from uranium fission products, were evaluated and corrections applied where necessary. The 39.7-meter deep section comprises of Lishi Loess of the middle Pleistocene (Q2) and Malan Loess of the late Pleistocene (Q3). The section is characterized by the presence of 5 layers of paleosol, and each paleosol is underlain by a precipitation layer. When the elemental abundances are converted to a carbonate-free basis, there is little compositional difference among the carbonate-free fractions of loess, paleosol and precipitation layers. This indicates that dissolution of carbonate minerals by downward-moving surface water was an important process in paleosol formation while other minerals were not severely weathered and elemental fractionation was minimal. The parent materials of the paleosol and precipitation layers closely resemble the loess layers in their elemental abundances, which suggests that all layers in the section have a compositionally similar source. ?? 1987 Akade??miai Kiado??.

  9. Lipid and fatty acid compositions of cod ( Gadus morhua), haddock ( Melanogrammus aeglefinus) and halibut ( Hippoglossus hippoglossus)

    NASA Astrophysics Data System (ADS)

    Zeng, Duan; Mai, Kangsen; Ai, Qinghui; Milley, Joyce E.; Lall, Santosh P.

    2010-12-01

    This study was conducted to compare lipid and fatty acid composition of cod, haddock and halibut. Three groups of cod (276 g ± 61 g), haddock (538 g ± 83 g) and halibut (3704 g ± 221 g) were maintained with commercial feeds mainly based on fish meal and marine fish oil for 12 weeks prior to sampling. The fatty acid compositions of muscle and liver were determined by GC/FID after derivatization of extracted lipids into fatty acid methyl esters (FAME). Lipids were also fractionated into neutral and polar lipids using Waters silica Sep-Pak?. The phospholipid fraction was further separated by high-performance thin-layer chromatography (HPTLC) and the FAME profile was obtained. Results of the present study showed that cod and haddock were lean fish and their total muscle lipid contents were 0.8% and 0.7%, respectively, with phospholipid constituting 83.6% and 87.5% of the total muscle lipid, respectively. Halibut was a medium-fat fish and its muscle lipid content was 8%, with 84% of the total muscle lipid being neutral lipid. Total liver lipid contents of cod, haddock and halibut were 36.9%, 67.2% and 30.7%, respectively, of which the neutral lipids accounted for the major fraction (88.1%-97.1%). Polyunsaturated fatty acids were the most abundant in cod and haddock muscle neutral lipid. Monounsaturated fatty acid level was the highest in halibut muscle neutral lipid. Fatty acid compositions of phospholipid were relatively constant. In summary, the liver of cod and haddock as lean fish was the main lipid reserve organ, and structural phospholipid is the major lipid form in flesh. However, as a medium-fat fish, halibut stored lipid in both their liver and muscle.

  10. Note: Accurate determination of thickness of multiple layers of thin film deposited on a piezoelectric quartz crystal.

    PubMed

    Wajid, Abdul

    2013-10-01

    Modern day piezoelectric quartz crystal microbalances for thin film deposition control are based on Z-match equation, which is mathematically valid for deposition of a single material on a given quartz crystal. When multiple layers are deposited, thickness and deposition rate errors accumulate due to mismatch of acoustic impedance of different materials. Here we present a novel method, based on the acoustic transfer matrix formalism, for accurate determination of thickness of an arbitrary number of layers of dissimilar materials deposited on a quartz crystal. Laboratory data show excellent accuracy of the method compared to conventional Z-match equation. PMID:24182174

  11. Improvement of calibration curve for determining peroxide values of food lipids by the modified ferrous oxidation-xylenol orange method.

    PubMed

    Burat, K M; Bozkurt, O

    1996-01-01

    The linearity range of the standard Fe(+3) calibration curve for measuring lipid hydroperoxides in food samples by the modified ferrous oxidation-xylenol orange (mFOX) method was extended from 5-20 micrograms to 2-40 micrograms by establishing the best concentration of xylenol orange in the same assay system. Butter, fish oil, and some vegetable oils were analyzed by the International Dairy Federation method and the improved mFOX method, and results were compared.

  12. Separation of Intra- and Extramyocellular Lipid Signals in Proton MR Spectra by Determination of Their Magnetic Field Distribution

    NASA Astrophysics Data System (ADS)

    Steidle, G.; Machann, J.; Claussen, C. D.; Schick, F.

    2002-02-01

    In skeletal musculature intramyocellular (IMCL) and extramyocellular lipids (EMCL) are stored in compartments of different geometry and experience different magnetic field strengths due to geometrical susceptibility effects. The effect is strong enough to-at least partly-separate IMCL and EMCL contributions in 1H MR spectroscopy, despite IMCL and EMCL consisting of the same substances. The assessment of intramyocellular lipid stores in skeletal musculature by 1H MR spectroscopy plays an important role for studying physiological and pathological aspects of lipid metabolism. Therefore, a method using mathematical tools of Fourier analysis is developed to obtain the magnetic field distribution (MFD) from the measured spectra by deconvolution. A reference lipid spectrum is required which was recorded in tibial yellow bone marrow. It is shown that the separation of IMCL contributions can be performed more precisely-compared to other methods-based on the MFD. Examples of deconvolution in model systems elucidate the principle. Applications of the proposed approach on in vivo examinations in m. soleus and m. tibialis anterior are presented. Fitting the IMCL part of the MFD by a Gaussian lineshape with a linewidth kept fixed with respect to the linewidth of creatine and with the assumption of a smooth but not necessarily symmetrical shape for the EMCL part, the only free fit parameter, the amplitude of the IMCL part, is definite and subtraction leads to the EMCL part in the MFD. This procedure is especially justified for the soleus muscle showing a severely asymmetrical distribution which might lead to a marked overestimation of IMCL using common line fitting procedures.

  13. Microalgae lipid characterization.

    PubMed

    Yao, Linxing; Gerde, Jose A; Lee, Show-Ling; Wang, Tong; Harrata, Kamel A

    2015-02-18

    To meet the growing interest of utilizing microalgae biomass in the production of biofuels and nutraceutical and pharmaceutical lipids, we need suitable analytical methods and a comprehensive database for their lipid components. The objective of the present work was to demonstrate methodology and provide data on fatty acid composition, lipid class content and composition, characteristics of the unsaponifiables, and type of chlorophylls of five microalgae. Microalgae lipids were fractionated into TAG, FFA, and polar lipids using TLC, and the composition of fatty acids in total lipids and in each lipid class, hydrocarbons, and sterols were determined by GC-MS. Glyco- and phospholipids were profiled by LC/ESI-MS. Chlorophylls and their related metabolites were qualified by LC/APCI-MS. The melting and crystallization profiles of microalgae total lipids and their esters were analyzed by DSC to evaluate their potential biofuel applications. Significant differences and complexities of lipid composition among the algae tested were observed. The compositional information is valuable for strain selection, downstream biomass fractionation, and utilization.

  14. Multifunctional lipid multilayer stamping.

    PubMed

    Nafday, Omkar A; Lowry, Troy W; Lenhert, Steven

    2012-04-10

    Nanostructured lipid multilayers on surfaces are a promising biofunctional nanomaterial. For example, surface-supported lipid multilayer diffraction gratings with optical properties that depend on the microscale spacing of the grating lines and the nanometer thickness of the lipid multilayers have been fabricated previously by dip-pen nanolithography (DPN), with immediate applications as label-free biosensors. The innate biocompatibility of such gratings makes them promising as biological sensor elements, model cellular systems, and construction materials for nanotechnology. Here a method is described that combines the lateral patterning capabilities and scalability of microcontact printing with the topographical control of nanoimprint lithography and the multimaterial integration aspects of dip-pen nanolithography in order to create nanostructured lipid multilayer arrays. This approach is denoted multilayer stamping. The distinguishing characteristic of this method is that it allows control of the lipid multilayer thickness, which is a crucial nanoscale dimension that determines the optical properties of lipid multilayer nanostructures. The ability to integrate multiple lipid materials on the same surface is also demonstrated by multi-ink spotting onto a polydimethoxysilane stamp, as well as higher-throughput patterning (on the order of 2 cm(2) s(-1) for grating fabrication) and the ability to pattern lipid materials that could not previously be patterned with high resolution by lipid DPN, for example, the gel-phase phospholipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or the steroid cholesterol. PMID:22307810

  15. Determination of lipid-bound sulfate by ion chromatography and its application to quantification of sulfolipids from kidneys of various mammalian species.

    PubMed

    Tadano-Aritomi, K; Hikita, T; Suzuki, A; Toyoda, H; Toida, T; Imanari, T; Ishizuka, I

    2001-10-01

    A variety of procedures have been developed for determining the sulfate ester content of various biomolecules. Ion chromatography (IC), that is, quantitation of ionic substances by ion conductimetry after separation by anion-exchange chromatography, has been increasingly utilized for the determination of inorganic sulfate in clinical and environmental samples. We adopted suppressed-mode IC to the determination of lipid- or glycolipid-bound sulfate released by acid hydrolysis and found that it has the advantage of increased precision for wide concentration ranges (30 pmol to approximately micromol) and lack of interference from other lipids. To minimize deterioration of the separation column, the lipophilic constituents in the acid hydrolysate were removed by a two-phase partition system of chloroform-methanol-water. The inorganic sulfate was quantitatively extracted into the aqueous phase by replacing water with an alkaline buffer. By this method, the concentration of sulfolipids was determined in the kidney of mammals with various body mass. Sulfolipids were more concentrated in the kidney of smaller animals, which have higher maximum urine concentrating activity per gram of body mass, supporting the hypothesis of the function of sulfolipids as an ion barrier on the luminal surface of renal tubules.

  16. A novel human erythrocyte glycosylphosphatidylinositol (GPI)-anchored glycoprotein ACA. Isolation, purification, primary structure determination, and molecular parameters of its lipid structure.

    PubMed

    Becker Kojić, Zorica A; Terness, Peter

    2002-10-25

    A method has been elaborated to isolate and purify up to homogeneity a novel membrane glycoprotein containing a glycosyl-phosphatidylinositol (GPI) anchor by means of salting out with ammonium sulfate (40-80% saturation), followed by preparative SDS-PAGE, chromatography and acetone precipitation. The preparation obtained was homogeneous upon electrophoresis in the presence of 0.1% SDS after reduction with 2-mercaptoethanol. It is protein-soluble at its isoelectrical point (pH 5.5) with molecular mass of 65,000 daltons. The isolated protein is linked to the membrane via glycosyl-phosphatidylinositol susceptible to cleavage by purified phospholipase C. The hydrophobic portion of the glycolipid membrane anchor of the protein was radiolabeled with the photoactivated reagent 3-(trifluoromethyl)-3-(m-[(125)I]iodophenyl)diazirine and hydrolyzed with glycosyl-phosphatidylinositol-specific phospholipase C, followed by enzymatic deacetylation of the remaining lipid. Thin-layer chromatography showed that the generated radiolabeled fragment migrates with the same mobility as that of variant surface glycoprotein (VSG), obtained in the same manner. In this study we describe a novel erythrocyte membrane GPI-linked protein with the structural feature of an anchor that, in contrast to other GPI-linked erythrocyte proteins, has a non-acetylated inositol ring and diacylglycerol rather than alkyl-acyl glycerol as a lipid tail of the anchor.

  17. The internal meristem layer (L3) determines floral meristem size and carpel number in tomato periclinal chimeras.

    PubMed

    Szymkowiak, E J; Sussex, I M

    1992-09-01

    Cell-cell interactions are important during plant development. We have generated periclinal chimeras between plants that differ in the number of carpels per flower to determine the roles of cells occupying specific positions in the floral meristem in determining the number of carpels initiated. Intraspecific chimeras were generated between tomato (Lycopersicon esculentum) expressing the mutation fasciated, which causes an increased number of floral organs per whorl, and tomato wild type for fasciated. Interspecific chimeras were generated between tomato and L. peruvianum, which differ in number of carpels per flower. In both sets of chimeras, carpel number as well as the size of the floral meristem during carpel initiation were not determined by the genotype of cells in the outer two layers of the meristem (L1 and L2) but were determined by the genotype of cells occupying the inner layer (L3) of the meristem. We concluded from these experiments that during floral organ initiation, cells in certain layers of the meristem respond to information supplied to them from other cells in the meristem.

  18. Determination of the parameters of a holographic layer from its spectral characteristics

    NASA Astrophysics Data System (ADS)

    Kraiskii, A. A.; Kraiskii, A. V.

    2016-06-01

    Methods for estimating the main parameters of holographic sensors (refractive index modulation depth and hologram thickness) from transmission spectra in the absence of absorption and light scattering are discussed. The consideration is performed for layers oriented parallel to the holographic layer surface under normal light incidence. Direct numerical solution of the problem of light propagation in a periodic nonabsorbing medium is used to study the reflection and transmission spectra of the holographic layer in a wide range of variation in its thickness and the refractive index modulation depth. A classification of the reflection regimes from the holographic layer is proposed (from weak reflection to the photonic crystal regime). A comparison with the results obtained by the coupled-wave analysis is performed, and the limitations of this method at a significant spectral detuning from resonance and under conditions of strong reflection are revealed. It is shown that the main hologram parameters can be estimated from the experimental transmission spectrum of the phase hologram (in the case of strong reflection) based on the spectral dip parameters.

  19. TRPV1 structures in nanodiscs reveal mechanisms of ligand and lipid action.

    PubMed

    Gao, Yuan; Cao, Erhu; Julius, David; Cheng, Yifan

    2016-06-16

    When integral membrane proteins are visualized in detergents or other artificial systems, an important layer of information is lost regarding lipid interactions and their effects on protein structure. This is especially relevant to proteins for which lipids have both structural and regulatory roles. Here we demonstrate the power of combining electron cryo-microscopy with lipid nanodisc technology to ascertain the structure of the rat TRPV1 ion channel in a native bilayer environment. Using this approach, we determined the locations of annular and regulatory lipids and showed that specific phospholipid interactions enhance binding of a spider toxin to TRPV1 through formation of a tripartite complex. Furthermore, phosphatidylinositol lipids occupy the binding site for capsaicin and other vanilloid ligands, suggesting a mechanism whereby chemical or thermal stimuli elicit channel activation by promoting the release of bioactive lipids from a critical allosteric regulatory site. PMID:27281200

  20. Summary of experimentally determined facts concerning the behavior of the boundary layer and performance of boundary layer measurements. [considering sailing flight

    NASA Technical Reports Server (NTRS)

    Vanness, W.

    1978-01-01

    A summary report of boundary layer studies is presented. Preliminary results of experimental measurements show that: (1) A very thin layer (approximately 0.4 mm) of the boundary layer seems to be accelerated; (2) the static pressure of the outer flow does not remain exactly constant through the boundary layer; and (3) an oncoming boundary layer which is already turbulent at the suction point can again become laminar behind this point without being completely sucked off.

  1. Membrane location of apocytochrome c and cytochrome c determined from lipid-protein spin exchange interactions by continuous wave saturation electron spin resonance.

    PubMed Central

    Snel, M M; Marsh, D

    1994-01-01

    Apocytochrome c derived from horse heart cytochrome c was spin-labeled on the cysteine residue at position 14 or 17 in the N-terminal region of the primary sequence, and cytochrome c from yeast was spin-labeled on the single cysteine residue at sequence position 102 in the C-terminal region. The spin-labeled apocytochrome c and cytochrome c were bound to fluid bilayers composed of different negatively charged phospholipids that also contained phospholipid probes that were spin-labeled either in the headgroup or at different positions in the sn-2 acyl chain. The location of the spin-labeled cysteine residues on the lipid-bound proteins was determined relative to the spin-label positions in the different spin-labeled phospholipids by the influence of spin-spin interactions on the microwave saturation properties of the spin-label electron spin resonance spectra. The enhanced spin relaxation observed in the doubly labeled systems arises from Heisenberg spin exchange, which is determined by the accessibility of the spin-label group on the protein to that on the lipid. It is found that the labeled cysteine groups in horse heart apocytochrome c are located closest to the 14-C atom of the lipid acyl chain when the protein is bound to dimyristoyl- or dioleoyl-phosphatidylglycerol, and to that of the 5-C atom when the protein is bound to a dimyristoylphosphatidylglycerol/dimyristoylphosphatidylcholine (15:85 mol/mol mixture. On binding to dioleoylphosphatidylglycerol, the labeled cysteine residue in yeast cytochrome c is located closest to the phospholipid headgroups but possibly between the polar group region and the 5-C atom of the acyl chains. These data determine the extent to which the different regions of the proteins are able to penetrate negatively charged phospholipid bilayers. Images FIGURE 1 PMID:7948687

  2. Genetic determinants of long-term changes in blood lipid concentrations: 10-year follow-up of the GLACIER study.

    PubMed

    Varga, Tibor V; Sonestedt, Emily; Shungin, Dmitry; Koivula, Robert W; Hallmans, Göran; Escher, Stefan A; Barroso, Inês; Nilsson, Peter; Melander, Olle; Orho-Melander, Marju; Renström, Frida; Franks, Paul W

    2014-06-01

    Recent genome-wide meta-analyses identified 157 loci associated with cross-sectional lipid traits. Here we tested whether these loci associate (singly and in trait-specific genetic risk scores [GRS]) with longitudinal changes in total cholesterol (TC) and triglyceride (TG) levels in a population-based prospective cohort from Northern Sweden (the GLACIER Study). We sought replication in a southern Swedish cohort (the MDC Study; N = 2,943). GLACIER Study participants (N = 6,064) were genotyped with the MetaboChip array. Up to 3,495 participants had 10-yr follow-up data available in the GLACIER Study. The TC- and TG-specific GRSs were strongly associated with change in lipid levels (β = 0.02 mmol/l per effect allele per decade follow-up, P = 2.0 × 10(-11) for TC; β = 0.02 mmol/l per effect allele per decade follow-up, P = 5.0 × 10(-5) for TG). In individual SNP analysis, one TC locus, apolipoprotein E (APOE) rs4420638 (β = 0.12 mmol/l per effect allele per decade follow-up, P = 2.0 × 10(-5)), and two TG loci, tribbles pseudokinase 1 (TRIB1) rs2954029 (β = 0.09 mmol/l per effect allele per decade follow-up, P = 5.1 × 10(-4)) and apolipoprotein A-I (APOA1) rs6589564 (β = 0.31 mmol/l per effect allele per decade follow-up, P = 1.4 × 10(-8)), remained significantly associated with longitudinal changes for the respective traits after correction for multiple testing. An additional 12 loci were nominally associated with TC or TG changes. In replication analyses, the APOE rs4420638, TRIB1 rs2954029, and APOA1 rs6589564 associations were confirmed (P ≤ 0.001). In summary, trait-specific GRSs are robustly associated with 10-yr changes in lipid levels and three individual SNPs were strongly associated with 10-yr changes in lipid levels.

  3. Genetic Determinants of Long-Term Changes in Blood Lipid Concentrations: 10-Year Follow-Up of the GLACIER Study

    PubMed Central

    Varga, Tibor V.; Sonestedt, Emily; Shungin, Dmitry; Koivula, Robert W.; Hallmans, Göran; Escher, Stefan A.; Barroso, Inês; Nilsson, Peter; Melander, Olle; Orho-Melander, Marju; Renström, Frida; Franks, Paul W.

    2014-01-01

    Recent genome-wide meta-analyses identified 157 loci associated with cross-sectional lipid traits. Here we tested whether these loci associate (singly and in trait-specific genetic risk scores [GRS]) with longitudinal changes in total cholesterol (TC) and triglyceride (TG) levels in a population-based prospective cohort from Northern Sweden (the GLACIER Study). We sought replication in a southern Swedish cohort (the MDC Study; N = 2,943). GLACIER Study participants (N = 6,064) were genotyped with the MetaboChip array. Up to 3,495 participants had 10-yr follow-up data available in the GLACIER Study. The TC- and TG-specific GRSs were strongly associated with change in lipid levels (β = 0.02 mmol/l per effect allele per decade follow-up, P = 2.0×10−11 for TC; β = 0.02 mmol/l per effect allele per decade follow-up, P = 5.0×10−5 for TG). In individual SNP analysis, one TC locus, apolipoprotein E (APOE) rs4420638 (β = 0.12 mmol/l per effect allele per decade follow-up, P = 2.0×10−5), and two TG loci, tribbles pseudokinase 1 (TRIB1) rs2954029 (β = 0.09 mmol/l per effect allele per decade follow-up, P = 5.1×10−4) and apolipoprotein A-I (APOA1) rs6589564 (β = 0.31 mmol/l per effect allele per decade follow-up, P = 1.4×10−8), remained significantly associated with longitudinal changes for the respective traits after correction for multiple testing. An additional 12 loci were nominally associated with TC or TG changes. In replication analyses, the APOE rs4420638, TRIB1 rs2954029, and APOA1 rs6589564 associations were confirmed (P≤0.001). In summary, trait-specific GRSs are robustly associated with 10-yr changes in lipid levels and three individual SNPs were strongly associated with 10-yr changes in lipid levels. PMID:24922540

  4. [Trigonelline in coffee. I. Comparison of thin layer chromatography with high-performance liquid chromatography. Simultaneous determination of caffeine].

    PubMed

    Stennert, A; Maier, H G

    1993-05-01

    Determination of trigonelline in several green and roasted coffees by thin-layer (TLC) and high-performance liquid chromatography (HPLC) were compared. Using TLC, two methods of detection were also compared. In addition, caffeine could be determined using HPLC. The determinations were each made with several variations of extraction and clean-up. Recoveries, variation coefficients and detection limits are listed. The most suitable method was HPLC in connection with hot water extraction. Only water is required as the mobile phase. The values for precision are in most cases better than (trigonelline) or similar to (caffeine) those reported in the literature. The determination of trigonelline by TLC, followed by extraction and photometry, is possible with a similar precision, but the simultaneous determination of caffeine was not tested. PMID:8511975

  5. Langmuir films study on lipid-containing artificial tears.

    PubMed

    Torrent-Burgués, J

    2016-04-01

    Lipid-containing artificial tears are a type of artificial tears that use lipid components in order to restore the lipid layer of the tear film. One of these components is lecithin which can be applied in spray solutions containing lecithin liposomes. In this work the behavior of three of these commercial tears based on lecithin, Innoxa, Opticalm and Optrex, are studied using the Langmuir technique. The obtained isotherms are presented, discussed and compared. This technique seems useful in order to see the film behavior of the lipid components of these tears and determine some important parameters such as fluidity and extension at the air-water interface, and allows us to discern differences between these commercial tears. Innoxa and Optrex tears are more similar to each other than to Opticalm tears. Opticalm presents more fluidity, probably due to the presence of more insaturations in the fatty acid chains of the phospholipids of the lecithin used in fabrication.

  6. Nonlinear physical segmentation algorithm for determining the layer boundary from lidar signal.

    PubMed

    Mao, Feiyue; Li, Jun; Li, Chen; Gong, Wei; Min, Qilong; Wang, Wei

    2015-11-30

    Layer boundary (base and top) detection is a basic problem in lidar data processing, the results of which are used as inputs of optical properties retrieval. However, traditional algorithms not only require manual intervention but also rely heavily on the signal-to-noise ratio. Therefore, we propose a robust and automatic algorithm for layer detection based on a novel algorithm for lidar signal segmentation and representation. Our algorithm is based on the lidar equation and avoids most of the limitations of the traditional algorithms. Testing of the simulated and real signals shows that the algorithm is able to position the base and top accurately even with a low signal to noise ratio. Furthermore, the results of the classification are accurate and satisfactory. The experimental results confirm that our algorithm can be used for automatic detection, retrieval, and analysis of lidar data sets.

  7. Carrier lifetime in exfoliated few-layer graphene determined from intersubband optical transitions.

    PubMed

    Limmer, Thomas; Feldmann, Jochen; Da Como, Enrico

    2013-05-24

    We report a femtosecond transient spectroscopy study in the near to middle infrared range, 0.8-0.35 eV photon energy, on graphene and few layer graphene single flakes. The spectra show an evolving structure of photoinduced absorption bands superimposed on the bleaching caused by Pauli blocking of the interband optically coupled states. Supported by tight-binding model calculations, we assign the photoinduced absorption features to intersubband transitions as the number of layers is increased. Interestingly, the intersubband photoinduced resonances show a longer dynamics than the interband bleaching, because of their independence from the absolute energy of the carriers with respect to the Dirac point. The dynamic of these intersubband transitions reflects the lifetime of the hot carriers and provides an elegant method to access it in this important class of semimetals.

  8. Carrier Lifetime in Exfoliated Few-Layer Graphene Determined from Intersubband Optical Transitions

    NASA Astrophysics Data System (ADS)

    Limmer, Thomas; Feldmann, Jochen; Da Como, Enrico

    2013-05-01

    We report a femtosecond transient spectroscopy study in the near to middle infrared range, 0.8-0.35 eV photon energy, on graphene and few layer graphene single flakes. The spectra show an evolving structure of photoinduced absorption bands superimposed on the bleaching caused by Pauli blocking of the interband optically coupled states. Supported by tight-binding model calculations, we assign the photoinduced absorption features to intersubband transitions as the number of layers is increased. Interestingly, the intersubband photoinduced resonances show a longer dynamics than the interband bleaching, because of their independence from the absolute energy of the carriers with respect to the Dirac point. The dynamic of these intersubband transitions reflects the lifetime of the hot carriers and provides an elegant method to access it in this important class of semimetals.

  9. Lipids in the walls and contents of jaw cysts.

    PubMed

    Stokke, T

    1976-11-01

    The amount and composition of lipid in 129 walls of jaw cyst and 18 pieces of cyst contents was determined by thin-layer chromatography. The proportions of the fractions (i.e. phospholipid, free cholesterol, cholesteryl ester, triglyceride, free fatty acids) were compared with corresponding values (collected from the literature) from the superficial layers of normal and atherosclerotic aortas. The amounts of total lipid and of the fractions were comparable. Concomitant with an increasing amount of lipid, there was a rise in the ratio: total cholesterol/phospholipid. In the aortic lesions this increase of cholesterol was due to a rise in the fraction of cholesteryl ester. In the cyst material, the increase was due to a rise in the free cholesterol. A possible explanation is discussed.

  10. Theoretical Determination of The Optimum Thickness of Perylene Layer in Bilayer Phthalocyanine/Perylene Photovoltaic Device

    NASA Astrophysics Data System (ADS)

    Pratiwi, Herlina; Siahaan, Timothy; Satriawan, Mirza; Nurwantoro, Pekik; Triyana, Kuwat

    2009-09-01

    We do theoretical study on thickness of the active layers in a heterojunction bilayer thin film photovoltaic device based on copper phthalocyanine (CuPc)/perylene that gives the highest Incident Photon to Current Efficiency (IPCE). The device we study consists Glass (1 mm)/ITO (Indium Tin Oxide, 120 nm)/CuPc (50 nm)/PTCDA (3, 4, 9, 10-perylenetetracarboxylic dianhydride, x nm)/Ag (40 nm), where x is the thickness of the PTCDA layer that we calculate here. The calculation is based on assumption that the photocurrent generation process is the result of the creation of photogenerated excitons, which difuse before dissociated at the CuPc/PTCDA interface following the diffusion equation, by internal optical electric field that comes from light exposure. We also assume that almost all photocurrent is created in the CuPc/PTCDA interface. Because the order of the thickness of the active layers is the same or smaller than of the wavelength of visible light, we take into account the effect of reflection and interference in the calculation of internal optical electric field distribution inside the device by making use complex indices of refraction of the active materials in our calculation. The modulus of it is proportional with the number generated excitons. The general solution of the exciton diffusion equation was used for calculating the photocurrent and the IPCE. Here, we find the optimum thickness of PTCDA layer that gives greatest IPCE at the wavelength of 344 nm and 467 nm, which are the wavelengths at which the absorption coefficients of CuPc and PTCDA, respectively, reach the maximum values.

  11. Large-scale recrystallization of the S-layer of Bacillus coagulans E38-66 at the air/water interface and on lipid films.

    PubMed Central

    Pum, D; Weinhandl, M; Hödl, C; Sleytr, U B

    1993-01-01

    S-layer protein isolated from Bacillus coagulans E38-66 could be recrystallized into large-scale coherent monolayers at an air/water interface and on phospholipid films spread on a Langmuir-Blodgett trough. Because of the asymmetry in the physiochemical surface properties of the S-layer protein, the subunits were associated with their more hydrophobic outer face with the air/water interface and oriented with their negatively charged inner face to the zwitterionic head groups of the dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylethanolamine (DPPE) monolayer films. The dynamic crystal growth at both types of interfaces was first initiated at several distant nucleation points. The individual monocrystalline areas grew isotropically in all directions until the front edge of neighboring crystals was met. The recrystallized S-layer protein and the S-layer-DPPE layer could be chemically cross-linked from the subphase with glutaraldehyde. Images PMID:8478338

  12. Determination of monomethylmercury and dimethylmercury in the Arctic marine boundary layer.

    PubMed

    Baya, Pascale A; Gosselin, Michel; Lehnherr, Igor; St Louis, Vincent L; Hintelmann, Holger

    2015-01-01

    Our understanding of the biogeochemical cycling of monomethylmercury (MMHg) in the Arctic is incomplete because atmospheric sources and sinks of MMHg are still unclear. We sampled air in the Canadian Arctic marine boundary layer to quantify, for the first time, atmospheric concentrations of methylated Hg species (both MMHg and dimethylmercury (DMHg)), and, estimate the importance of atmospheric deposition as a source of MMHg to Arctic land- and sea-scapes. Overall atmospheric MMHg and DMHg concentrations (mean ± SD) were 2.9 ± 3.6 and 3.8 ± 3.1 (n = 37) pg m(-3), respectively. Concentrations of methylated Hg species in the marine boundary layer varied significantly among our sites, with a predominance of MMHg over Hudson Bay (HB), and DMHg over Canadian Arctic Archipelago (CAA) waters. We concluded that DMHg is of marine origin and that primary production rate and sea-ice cover are major drivers of its concentration in the Canadian Arctic marine boundary layer. Summer wet deposition rates of atmospheric MMHg, likely to be the product of DMHg degradation in the atmosphere, were estimated at 188 ± 117.5 ng m(-2) and 37 ± 21.7 ng m(-2) for HB and CAA, respectively, sustaining MMHg concentrations available for biomagnification in the pelagic food web.

  13. Determination of the mutual coherence function and determination of the point-spread function in a transversely and longitudinally inhomogeneous aero-optic turbulence layer.

    PubMed

    Monteiro, A; Jarem, J

    1993-01-10

    The mutual coherence function (MCF) of strong-fluctuation theory as a result of optical energy passing through a transversely and longitudinally inhomogeneous aero-optic turbulent layer is studied. Solutions for the MCF equation are determined by decomposing the MCF solution into coherent and incoherent parts and by solving separately the equations that result from this decomposition. The MCF equations for an arbitrary three-dimensional inhomogeneous layer are presented. A simplified version of these equations for the case in which the turbulence inhomogeneity is longitudinally inhomogeneous and is transversely inhomogeneous in one dimension is also presented. A numerical method for solving the parabolic MCF equations by the Lax-Wendroff explicit finite-difference algorithm is given, and numerical examples of the MCF solution for three different inhomogeneous aero-optic layers are discussed. Equations to relate the point-spread function, the optical transfer function, and image formation to the MCF of an inhomogeneous aero-optic turbulence layer are derived. An approximate MCF Fourier integral solution is presented and compared with the exact finite-difference solution. A formula to estimate the validity of the approximate integral solution is given. PMID:20802679

  14. Configuration of PKCα-C2 Domain Bound to Mixed SOPC/SOPS Lipid Monolayers

    PubMed Central

    Chen, Chiu-Hao; Málková, Šárka; Pingali, Sai Venkatesh; Long, Fei; Garde, Shekhar; Cho, Wonhwa; Schlossman, Mark L.

    2009-01-01

    Abstract X-ray reflectivity measurements are used to determine the configuration of the C2 domain of protein kinase Cα (PKCα-C2) bound to a lipid monolayer of a 7:3 mixture of 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine and 1-stearoyl-2-oleoyl-sn-glycero-3-phosphoserine supported on a buffered aqueous solution. The reflectivity is analyzed in terms of the known crystallographic structure of PKCα-C2 and a slab model representation of the lipid layer. The configuration of lipid-bound PKCα-C2 is described by two angles that define its orientation, θ = 35° ± 10° and ϕ =210° ± 30°, and a penetration depth (=7.5 ± 2 Å) into the lipid layer. In this structure, the β-sheets of PKCα-C2 are nearly perpendicular to the lipid layer and the domain penetrates into the headgroup region of the lipid layer, but not into the tailgroup region. This configuration of PKCα-C2 determined by our x-ray reflectivity is consistent with many previous findings, particularly mutational studies, and also provides what we believe is new molecular insight into the mechanism of PKCα enzyme activation. Our analysis method, which allows us to test all possible protein orientations, shows that our data cannot be explained by a protein that is orientated parallel to the membrane, as suggested by earlier work. PMID:19917234

  15. Harvesting and cryo-cooling crystals of membrane proteins grown in lipidic mesophases for structure determination by macromolecular crystallography.

    PubMed

    Li, Dianfan; Boland, Coilín; Aragao, David; Walsh, Kilian; Caffrey, Martin

    2012-09-02

    An important route to understanding how proteins function at a mechanistic level is to have the structure of the target protein available, ideally at atomic resolution. Presently, there is only one way to capture such information as applied to integral membrane proteins (Figure 1), and the complexes they form, and that method is macromolecular X-ray crystallography (MX). To do MX diffraction quality crystals are needed which, in the case of membrane proteins, do not form readily. A method for crystallizing membrane proteins that involves the use of lipidic mesophases, specifically the cubic and sponge phases(1-5), has gained considerable attention of late due to the successes it has had in the G protein-coupled receptor field(6-21) (www.mpdb.tcd.ie). However, the method, henceforth referred to as the in meso or lipidic cubic phase method, comes with its own technical challenges. These arise, in part, due to the generally viscous and sticky nature of the lipidic mesophase in which the crystals, which are often micro-crystals, grow. Manipulating crystals becomes difficult as a result and particularly so during harvesting(22,23). Problems arise too at the step that precedes harvesting which requires that the glass sandwich plates in which the crystals grow (Figure 2)(24,25) are opened to expose the mesophase bolus, and the crystals therein, for harvesting, cryo-cooling and eventual X-ray diffraction data collection. The cubic and sponge mesophase variants (Figure 3) from which crystals must be harvested have profoundly different rheologies(4,26). The cubic phase is viscous and sticky akin to a thick toothpaste. By contrast, the sponge phase is more fluid with a distinct tendency to flow. Accordingly, different approaches for opening crystallization wells containing crystals growing in the cubic and the sponge phase are called for as indeed different methods are required for harvesting crystals from the two mesophase types. Protocols for doing just that have been

  16. Employment of Mixed Layer Models and Large Eddy Simulations to Determine the Factors Controlling Stratocumulus Cloud Lifetime over the Coast

    NASA Astrophysics Data System (ADS)

    Ghonima, M. S.; Heus, T.; Norris, J. R.; Kleissl, J. P.

    2015-12-01

    Summertime marine boundary layer stratocumulus (Sc) clouds have a strong impact on ecology and infrastructure over the coast of California. Modeling the lifetime of such clouds in global climate models (GCM) or numerical weather prediction models (NWP) is difficult and significant errors are typically observed. For instance, stratocumulus clouds over the coast of southern California in the Weather Research and Forecasting (WRF) model were found to dissipate, on average, 1.9 hours earlier than observed via satellite. In order to determine the factors controlling the Sc lifetime, we have employed large eddy simulations (LES) and a mixed layer model (MLM). Enhancements to previous MLMs include a temperature dependent radiation scheme, a land surface model, and a novel entrainment parameterization scheme for stratocumulus clouds over land in which the entrainment velocity is derived as a function of the surface buoyancy flux and the buoyancy flux integrated over the cloud layer. The advantage of using the MLM is that different mechanisms and feedbacks controlling stratocumulus cloud thickness can be examined rapidly through sensitivity studies. We find that during the night cloud lifetime is modulated by longwave cooling of the boundary layer and entrainment flux warming and drying. During the day, surface shortwave radiative heating drives surface flux therefore increasing the turbulence within the boundary layer and increasing entrainment flux. For wet surface conditions, the increase in latent heat flux moistens the boundary layer and offsets the increase in entrainment flux warming and drying of the boundary layer and clouds persist throughout the day. For dry surface conditions, the combination of increased surface sensible heat flux warming the boundary layer and increased entrainment flux act to dissipate the cloud within a couple of hours after sunrise. For both cases, the sea breeze advects cool ocean air that acts to thicken and prolong the cloud lifetime

  17. Measurements and determination of the marine coarse aerosol fluxes in near marine boundary layer.

    NASA Astrophysics Data System (ADS)

    Markuszewski, Piotr; Petelski, Tomasz; Zielinski, Tymon; Strzalkowska, Agata; Pakszys, Paulina; Makuch, Przemyslaw; Kowalczyk, Jakub

    2014-05-01

    Studies of production and transport of aerosol over the sea are very important for many areas of knowledge. Marine aerosols emitted from the sea surface help to clean the boundary layer from other aerosol particles. The emitted droplets do not dry out in the highly humid surface layer air and because of their sizes most of them are deposited quickly at the sea surface. Therefore, marine aerosols have many features of rain i.e. the deposition in the marine boundary layer in high wind events is controlled not only by the "dry" processes but also by the "wet" scavenging. During a number of cruises conducted on board of r/v Oceania between 2008 and 2012 we collected much data which were further used to calculate sea salt source function over the Baltic Sea. Measurements were carried out using a gradient method. For this method we used a Laser Particle Counter (PMS model CSASP-100_HV) placed on one of the masts of the boat. Measurements were performed at five different levels above the sea level: 8, 11, 14, 17 and 20 meters. The vertical aerosol concentration gradient was obtained from a minimum of 4 measurement series. Thus each result consists of a 1 hour series with the average sampling time at each elevation equaling to 8 minutes. Based on the averaged vertical concentration, and using the Monin Obukhov theory, profiles of vertical sea spray fluxes in the near water layer were calculated. Using the results from those experiments the sea spray emission fluxes have been calculated for all particles of sizes at ranges from 0.5 μm to 8 μm, as well as for particles of sizes from fifteen channels of 0.5 μm width. Using these fluxes we calculated the Sea Salt Generation Function (SSGF) over the Baltic Sea. This function provides information on the emission of particles of different sizes, depending on environmental parameters. The emission of sea spray depends on the magnitude of energy lost by the wind waves in the process of their collapse. The support for this study

  18. Aliphatic chain length by isotropic mixing (ALCHIM): determining composition of complex lipid samples by 1H NMR spectroscopy

    PubMed Central

    Yi, Ruiyang; Volden, Paul A.; Conzen, Suzanne D.

    2015-01-01

    Quantifying the amounts and types of lipids present in mixtures is important in fields as diverse as medicine, food science, and biochemistry. Nuclear magnetic resonance (NMR) spectroscopy can quantify the total amounts of saturated and unsaturated fatty acids in mixtures, but identifying the length of saturated fatty acid or the position of unsaturation by NMR is a daunting challenge. We have developed an NMR technique, aliphatic chain length by isotropic mixing, to address this problem. Using a selective total correlation spectroscopy technique to excite and transfer magnetization from a resolved resonance, we demonstrate that the time dependence of this transfer to another resolved site depends linearly on the number of aliphatic carbons separating the two sites. This technique is applied to complex natural mixtures allowing the identification and quantification of the constituent fatty acids. The method has been applied to whole adipocytes demonstrating that it will be of great use in studies of whole tissues. PMID:24831341

  19. Conduction band offset determination between strained CdSe and ZnSe layers using DLTS

    SciTech Connect

    Rangel-Kuoppa, Victor-Tapio

    2013-12-04

    The conduction band offset between strained CdSe layers embedded in unintentionally n-type doped ZnSe is measured and reported. Two samples, consisting of thirty Ultra Thin Quantum Wells (UTQWs) of CdSe embedded in ZnSe, grown by Atomic Layer Epitaxy, are used for this study. The thicknesses of the UTQWs are one and three monolayers (MLs) in each sample, respectively. As expected, the sample with one ML UTQWs does not show any energy level in the UTQWs due to the small thickness of the UTQWs, while the thickness of the sample with 3 ML UTQWs is large enough to form an energy level inside the UTQWs. This energy level appears as a majority trap with an activation energy of 223.58 ± 9.54 meV. This corresponds to UTQWs with barrier heights (the conduction band offset) between 742 meV and 784 meV. These values suggest that the band gap misfit between strained CdSe and ZnSe is around 70.5 to 74 % in the conduction band.

  20. Determinants of carbon release from the active layer and permafrost deposits on the Tibetan Plateau

    PubMed Central

    Chen, Leiyi; Liang, Junyi; Qin, Shuqi; Liu, Li; Fang, Kai; Xu, Yunping; Ding, Jinzhi; Li, Fei; Luo, Yiqi; Yang, Yuanhe

    2016-01-01

    The sign and magnitude of permafrost carbon (C)-climate feedback are highly uncertain due to the limited understanding of the decomposability of thawing permafrost and relevant mechanistic controls over C release. Here, by combining aerobic incubation with biomarker analysis and a three-pool model, we reveal that C quality (represented by a higher amount of fast cycling C but a lower amount of recalcitrant C compounds) and normalized CO2–C release in permafrost deposits were similar or even higher than those in the active layer, demonstrating a high vulnerability of C in Tibetan upland permafrost. We also illustrate that C quality exerts the most control over CO2–C release from the active layer, whereas soil microbial abundance is more directly associated with CO2–C release after permafrost thaw. Taken together, our findings highlight the importance of incorporating microbial properties into Earth System Models when predicting permafrost C dynamics under a changing environment. PMID:27703168

  1. Satb2 is a postmitotic determinant for upper-layer neuron specification in the neocortex.

    PubMed

    Britanova, Olga; de Juan Romero, Camino; Cheung, Amanda; Kwan, Kenneth Y; Schwark, Manuela; Gyorgy, Andrea; Vogel, Tanja; Akopov, Sergey; Mitkovski, Miso; Agoston, Denes; Sestan, Nenad; Molnár, Zoltán; Tarabykin, Victor

    2008-02-01

    Pyramidal neurons of the neocortex can be subdivided into two major groups: deep- (DL) and upper-layer (UL) neurons. Here we report that the expression of the AT-rich DNA-binding protein Satb2 defines two subclasses of UL neurons: UL1 (Satb2 positive) and UL2 (Satb2 negative). In the absence of Satb2, UL1 neurons lose their identity and activate DL- and UL2-specific genetic programs. UL1 neurons in Satb2 mutants fail to migrate to superficial layers and do not contribute to the corpus callosum but to the corticospinal tract, which is normally populated by DL axons. Ctip2, a gene required for the formation of the corticospinal tract, is ectopically expressed in all UL1 neurons in the absence of Satb2. Satb2 protein interacts with the Ctip2 genomic region and controls chromatin remodeling at this locus. Satb2 therefore is required for the initiation of the UL1-specific genetic program and for the inactivation of DL- and UL2-specific genes. PMID:18255031

  2. Acyl-Lipid Metabolism

    PubMed Central

    Li-Beisson, Yonghua; Shorrosh, Basil; Beisson, Fred; Andersson, Mats X.; Arondel, Vincent; Bates, Philip D.; Baud, Sébastien; Bird, David; DeBono, Allan; Durrett, Timothy P.; Franke, Rochus B.; Graham, Ian A.; Katayama, Kenta; Kelly, Amélie A.; Larson, Tony; Markham, Jonathan E.; Miquel, Martine; Molina, Isabel; Nishida, Ikuo; Rowland, Owen; Samuels, Lacey; Schmid, Katherine M.; Wada, Hajime; Welti, Ruth; Xu, Changcheng; Zallot, Rémi; Ohlrogge, John

    2013-01-01

    Acyl lipids in Arabidopsis and all other plants have a myriad of diverse functions. These include providing the core diffusion barrier of the membranes that separates cells and subcellular organelles. This function alone involves more than 10 membrane lipid classes, including the phospholipids, galactolipids, and sphingolipids, and within each class the variations in acyl chain composition expand the number of structures to several hundred possible molecular species. Acyl lipids in the form of triacylglycerol account for 35% of the weight of Arabidopsis seeds and represent their major form of carbon and energy storage. A layer of cutin and cuticular waxes that restricts the loss of water and provides protection from invasions by pathogens and other stresses covers the entire aerial surface of Arabidopsis. Similar functions are provided by suberin and its associated waxes that are localized in roots, seed coats, and abscission zones and are produced in response to wounding. This chapter focuses on the metabolic pathways that are associated with the biosynthesis and degradation of the acyl lipids mentioned above. These pathways, enzymes, and genes are also presented in detail in an associated website (ARALIP: http://aralip.plantbiology.msu.edu/). Protocols and methods used for analysis of Arabidopsis lipids are provided. Finally, a detailed summary of the composition of Arabidopsis lipids is provided in three figures and 15 tables. PMID:23505340

  3. Acyl-Lipid Metabolism

    PubMed Central

    Li-Beisson, Yonghua; Shorrosh, Basil; Beisson, Fred; Andersson, Mats X.; Arondel, Vincent; Bates, Philip D.; Baud, Sébastien; Bird, David; DeBono, Allan; Durrett, Timothy P.; Franke, Rochus B.; Graham, Ian A.; Katayama, Kenta; Kelly, Amélie A.; Larson, Tony; Markham, Jonathan E.; Miquel, Martine; Molina, Isabel; Nishida, Ikuo; Rowland, Owen; Samuels, Lacey; Schmid, Katherine M.; Wada, Hajime; Welti, Ruth; Xu, Changcheng; Zallot, Rémi; Ohlrogge, John

    2010-01-01

    Acyl lipids in Arabidopsis and all other plants have a myriad of diverse functions. These include providing the core diffusion barrier of the membranes that separates cells and subcellular organelles. This function alone involves more than 10 membrane lipid classes, including the phospholipids, galactolipids, and sphingolipids, and within each class the variations in acyl chain composition expand the number of structures to several hundred possible molecular species. Acyl lipids in the form of triacylglycerol account for 35% of the weight of Arabidopsis seeds and represent their major form of carbon and energy storage. A layer of cutin and cuticular waxes that restricts the loss of water and provides protection from invasions by pathogens and other stresses covers the entire aerial surface of Arabidopsis. Similar functions are provided by suberin and its associated waxes that are localized in roots, seed coats, and abscission zones and are produced in response to wounding. This chapter focuses on the metabolic pathways that are associated with the biosynthesis and degradation of the acyl lipids mentioned above. These pathways, enzymes, and genes are also presented in detail in an associated website (ARALIP: http://aralip.plantbiology.msu.edu/). Protocols and methods used for analysis of Arabidopsis lipids are provided. Finally, a detailed summary of the composition of Arabidopsis lipids is provided in three figures and 15 tables. PMID:22303259

  4. Acyl-lipid metabolism.

    PubMed

    Li-Beisson, Yonghua; Shorrosh, Basil; Beisson, Fred; Andersson, Mats X; Arondel, Vincent; Bates, Philip D; Baud, Sébastien; Bird, David; Debono, Allan; Durrett, Timothy P; Franke, Rochus B; Graham, Ian A; Katayama, Kenta; Kelly, Amélie A; Larson, Tony; Markham, Jonathan E; Miquel, Martine; Molina, Isabel; Nishida, Ikuo; Rowland, Owen; Samuels, Lacey; Schmid, Katherine M; Wada, Hajime; Welti, Ruth; Xu, Changcheng; Zallot, Rémi; Ohlrogge, John

    2013-01-01

    Acyl lipids in Arabidopsis and all other plants have a myriad of diverse functions. These include providing the core diffusion barrier of the membranes that separates cells and subcellular organelles. This function alone involves more than 10 membrane lipid classes, including the phospholipids, galactolipids, and sphingolipids, and within each class the variations in acyl chain composition expand the number of structures to several hundred possible molecular species. Acyl lipids in the form of triacylglycerol account for 35% of the weight of Arabidopsis seeds and represent their major form of carbon and energy storage. A layer of cutin and cuticular waxes that restricts the loss of water and provides protection from invasions by pathogens and other stresses covers the entire aerial surface of Arabidopsis. Similar functions are provided by suberin and its associated waxes that are localized in roots, seed coats, and abscission zones and are produced in response to wounding. This chapter focuses on the metabolic pathways that are associated with the biosynthesis and degradation of the acyl lipids mentioned above. These pathways, enzymes, and genes are also presented in detail in an associated website (ARALIP: http://aralip.plantbiology.msu.edu/). Protocols and methods used for analysis of Arabidopsis lipids are provided. Finally, a detailed summary of the composition of Arabidopsis lipids is provided in three figures and 15 tables. PMID:23505340

  5. Understanding and Analyzing Meibomian Lipids-A Review

    PubMed Central

    Butovich, Igor A.; Ham, Bryan M.

    2009-01-01

    Purpose This review is intended to bring to the informed reader the current state of knowledge about meibomian lipids and the art for analyzing them. Methods At the forefront of any endeavor, there are controversies, and these, along with future directions in the field, are brought to the reader's attention. Results Function and anatomy of meibomian glands are briefly covered, giving insight into possible mechanisms for secretory controls. Anatomically, some anomalies in meibomian gland distribution of different species, such as whales versus dolphins, are presented, and, for the first time, the structure of the meibomian glands in a selection of marsupials is presented. In attempting to make the literature more accessible, lipid structure and nomenclature are described, and these structures are related to their possible effects on the physicochemical properties of meibomian lipids. The advantages and disadvantages of various collection and storage techniques are described, as well as how gas chromatography and combined HPLC and mass spectrometry coupled with fragmentation are currently enabling us to determine the nature of the lipids in very small samples. Conclusions This review extends to discussing the lipids in tears (as opposed to meibomian gland lipids) and briefly highlights new thoughts about the interactions between proteins of the tear film and meibomian lipids. A model that includes proteins in the outer layer of the tear film is also presented. This model is currently being critically analyzed by the ocular community. It concludes briefly by highlighting possible further areas of research in this area. PMID:18568877

  6. Determining the efficiency of subjecting finely dispersed emulsions to physical coagulation in a packed layer under turbulent conditions

    NASA Astrophysics Data System (ADS)

    Laptev, A. G.; Basharov, M. M.; Farakhova, A. I.

    2013-09-01

    The process through which small droplets contained in emulsions are physically coagulated on the surface of random packing elements is considered. The theory of turbulent migration of a finely dispersed phase is used for determining the coagulation efficiency. Expressions for calculating coagulation efficiency and turbulent transfer rate are obtained by applying models of a turbulent boundary layer. An example of calculating the enlargement of water droplets in hydrocarbon medium represented by a wide fraction of light hydrocarbons (also known as natural gas liquid) is given. The process flowchart of a system for removing petroleum products from effluent waters discharged from the Kazan TETs-1 cogeneration station is considered. Replacement of the mechanical filter by a thin-layer settler with a coagulator is proposed.

  7. EFFECT OF TEMPERATURE ON THE C ISOTOPIC VALUE OF MICROBIAL LIPIDS APPLIED TO DETERMINE C USAGE IN MICROBIAL COMMUNITIES

    EPA Science Inventory

    The combination of compound specific stable isotopic analysis with phospholipid fatty acid (PLFAS) analysis is useful in determining the source of organic carbon used by groups of a microbial community. Determination of the effect of certain environmental parameters is important ...

  8. Symbiodinium Genotypic and Environmental Controls on Lipids in Reef Building Corals

    PubMed Central

    Cooper, Timothy F.; Lai, Michael; Ulstrup, Karin E.; Saunders, Sandra M.; Flematti, Gavin R.; Radford, Ben; van Oppen, Madeleine J. H.

    2011-01-01

    Background Lipids in reef building corals can be divided into two classes; non-polar storage lipids, e.g. wax esters and triglycerides, and polar structural lipids, e.g. phospholipids and cholesterol. Differences among algal endosymbiont types are known to have important influences on processes including growth and the photobiology of scleractinian corals yet very little is known about the role of symbiont types on lipid energy reserves. Methodology/Principal Findings The ratio of storage lipid and structural lipid fractions of Scott Reef corals were determined by thin layer chromatography. The lipid fraction ratio varied with depth and depended on symbiont type harboured by two corals (Seriatopora hystrix and Pachyseris speciosa). S. hystrix colonies associated with Symbiodinium C1 or C1/C# at deep depths (>23 m) had lower lipid fraction ratios (i.e. approximately equal parts of storage and structural lipids) than those with Symbiodinium D1 in shallow depths (<23 m), which had higher lipid fraction ratios (i.e. approximately double amounts of storage relative to structural lipid). Further, there was a non-linear relationship between the lipid fraction ratio and depth for S. hystrix with a modal peak at ∼23 m coinciding with the same depth as the shift from clade D to C types. In contrast, the proportional relationship between the lipid fraction ratio and depth for P. speciosa, which exhibited high specificity for Symbiodinium C3 like across the depth gradient, was indicative of greater amounts of storage lipids contained in the deep colonies. Conclusions/Significance This study has demonstrated that Symbiodinium exert significant controls over the quality of coral energy reserves over a large-scale depth gradient. We conclude that the competitive advantages and metabolic costs that arise from flexible associations with divergent symbiont types are offset by energetic trade-offs for the coral host. PMID:21637826

  9. Modified shape of the Eiffel Tower determined for an atmospheric boundary-layer wind profile

    NASA Astrophysics Data System (ADS)

    Weidman, P. D.

    2009-06-01

    The design and construction of the Eiffel Tower was based, in part, on a uniform horizontal wind model giving 300 kg m-2 kinematic pressure acting on the surface of the tower. Eiffel received a patent for his method of construction that eliminates the need for diagonal trellis bars used to resist the moment of an oncoming wind. At the end of the 19th century boundary-layer theory, laminar or turbulent, was nonexistent. Now, however, models for atmospheric flow over rough landscapes are available, the simplest being a power-law distribution of velocity with height. In this paper we deduce the shape of the tower had Eiffel incorporated this information into the design and construction of his world famous tower. Moreover, we prove Eiffel's observation that the tower profile conforms to the moment distribution wrought by the wind.

  10. Organic membranes determine the pattern of the columnar prismatic layer of mollusc shells.

    PubMed

    Checa, Antonio G; Macías-Sánchez, Elena; Harper, Elizabeth M; Cartwright, Julyan H E

    2016-05-11

    The degree to which biological control is exercised compared to physical control of the organization of biogenic materials is a central theme in biomineralization. We show that the outlines of biogenic calcite domains with organic membranes are always of simple geometries, while without they are much more complex. Moreover, the mineral prisms enclosed within the organic membranes are frequently polycrystalline. In the prismatic layer of the mollusc shell, organic membranes display a dynamics in accordance with the von Neumann-Mullins and Lewis Laws for two-dimensional foam, emulsion and grain growth. Taken together with the facts that we found instances in which the crystals do not obey such laws, and that the same organic membrane pattern can be found even without the mineral infilling, our work indicates that it is the membranes, not the mineral prisms, that control the pattern, and the mineral enclosed within the organic membranes passively adjusts to the dynamics dictated by the latter. PMID:27147096

  11. Boundary layer transition determination for periodic and static flows using phase-averaged pressure data

    NASA Astrophysics Data System (ADS)

    Gardner, A. D.; Richter, K.

    2015-06-01

    A method of boundary layer transition measurement is presented for wind tunnel models instrumented with surface pressure taps. The measurement relies on taking a number of theoretically identical measurements at different times and then analysing the standard deviation of the pressures. Due to the slight unsteady movement of the transition position, a peak in the standard deviation of pressure is found at the transition position, and this is correlated with measurements of the transition position with an infrared camera and hot-film anemometers. In contrast to microphone measurements, it is shown that the transition detection works for data which have been low-pass filtered with a cut-off of 1 Hz. The application to static and dynamic transition measurements on static and periodically pitching helicopter rotor blade airfoils at Mach 0.3-0.5 is demonstrated.

  12. Organic membranes determine the pattern of the columnar prismatic layer of mollusc shells.

    PubMed

    Checa, Antonio G; Macías-Sánchez, Elena; Harper, Elizabeth M; Cartwright, Julyan H E

    2016-05-11

    The degree to which biological control is exercised compared to physical control of the organization of biogenic materials is a central theme in biomineralization. We show that the outlines of biogenic calcite domains with organic membranes are always of simple geometries, while without they are much more complex. Moreover, the mineral prisms enclosed within the organic membranes are frequently polycrystalline. In the prismatic layer of the mollusc shell, organic membranes display a dynamics in accordance with the von Neumann-Mullins and Lewis Laws for two-dimensional foam, emulsion and grain growth. Taken together with the facts that we found instances in which the crystals do not obey such laws, and that the same organic membrane pattern can be found even without the mineral infilling, our work indicates that it is the membranes, not the mineral prisms, that control the pattern, and the mineral enclosed within the organic membranes passively adjusts to the dynamics dictated by the latter.

  13. How oil properties and layer thickness determine the entrainment of spilled surface oil.

    PubMed

    Zeinstra-Helfrich, Marieke; Koops, Wierd; Murk, Albertinka J

    2016-09-15

    Viscosity plays an important role in dispersion of spilled surface oil, so does adding chemical dispersants. For seven different oil grades, entrainment rate and initial droplet size distribution were investigated using a plunging jet apparatus with coupled camera equipment and subsequent image analysis. We found that amount of oil entrained is proportional to layer thickness and largely independent of oil properties: A dispersant dose of 1:200 did not result in a significantly different entrainment rate compared to no dispersants. Oil viscosity had a minor to no influence on entrainment rate, until a certain threshold above which entrainment was impeded. The mean droplet size scales with the modified Weber number as described by Johansen. The obtained results can help improve dispersion algorithms in oil spill fate and transport models, to aid making an informed decision about application of dispersants.

  14. How oil properties and layer thickness determine the entrainment of spilled surface oil.

    PubMed

    Zeinstra-Helfrich, Marieke; Koops, Wierd; Murk, Albertinka J

    2016-09-15

    Viscosity plays an important role in dispersion of spilled surface oil, so does adding chemical dispersants. For seven different oil grades, entrainment rate and initial droplet size distribution were investigated using a plunging jet apparatus with coupled camera equipment and subsequent image analysis. We found that amount of oil entrained is proportional to layer thickness and largely independent of oil properties: A dispersant dose of 1:200 did not result in a significantly different entrainment rate compared to no dispersants. Oil viscosity had a minor to no influence on entrainment rate, until a certain threshold above which entrainment was impeded. The mean droplet size scales with the modified Weber number as described by Johansen. The obtained results can help improve dispersion algorithms in oil spill fate and transport models, to aid making an informed decision about application of dispersants. PMID:27345705

  15. Determination of pigments in colour layers on walls of some selected historical buildings using optical and scanning electron microscopy

    SciTech Connect

    Skapin, A. Sever Ropret, P. Bukovec, P.

    2007-11-15

    For successful restoration of painted walls and painted coloured finishing coats it is necessary to determine the composition of the original colour layers. Identification of the pigments used in The Cistercian Abbey of Sticna and The Manor of Novo Celje was carried out using optical and scanning electron microscopy. Selected samples of wall paintings were inspected by the combined application of an optical microscope and a low-vacuum Scanning Electron Microscope to determine their colour and structural features and to identify the position of individual pigment grains. Energy dispersive spectroscopy was used to determine the elemental distribution on selected surfaces and elemental composition of individual pigments. It was found that the most abundantly used pigments were iron oxide red, cinnabar, green earth, umber, calcium carbonate white, ultramarine, yellow ochre and carbon black. These identifications have allowed us to compare the use of various pigments in buildings from different historical periods.

  16. Isolation and Characterization of Chicken Yolk Vitelline Membrane Lipids Using Eggs Enriched With Conjugated Linoleic Acid.

    PubMed

    Shinn, Sara Elizabeth; Liyanage, Rohana; Lay, Jackson O; Proctor, Andrew

    2016-06-01

    The vitelline membrane (VM) encloses the chicken egg yolk, separating it from albumen. The VM weakens during storage, and dietary lipid modification significantly affects its strength. However, no studies have characterize the fatty acyl residue (FA) composition of the VM, and reports of VM isolation and quantified lipid content are inconsistent. Therefore, the objectives of this study were: (1) to develop a washing and isolation method that removes residual yolk from VM without damage; (2) to determine the FA and lipid composition of CLA-rich egg yolk VM, relative to controls; (3) to determine the effect of 20 days of refrigeration on VM FA and lipid composition. To determine VM FA and lipid composition, 36 hens received either a corn-soybean meal-based control diet ("Control"), or the Control supplemented with either 10 % soy oil ("Soy control"), or 10 % CLA-rich soy oil ("CLA") for 30 days. VM were analyzed the day of collection ("fresh"), or after 20 days of refrigeration ("refrigerated"). There were no differences in FA compositions of fresh and refrigerated membranes within a treatment. CLA-rich yolk VM contains CLA, greater SFA, and significantly greater DHA relative to controls. Direct MALDI-TOF-MS identified 15 phosphatidylcholines, three phosphatidylethanolamines, one sphingomyelin, and 15 triacylglycerols in VM. Lipid species that showed significant differences among egg types included nine phosphatidylcholines and six triacylglycerols. MALDI analysis indicated significant differences in nine lipid classes on the VM inner layer. After refrigeration, five lipid classes on the inner layer and seven lipid classes on the outer layer had statistically significant differences among VM types. PMID:27108035

  17. Sequential ordered fatty acid alpha oxidation and Delta9 desaturation are major determinants of lipid storage and utilization in differentiating adipocytes.

    PubMed

    Su, Xiong; Han, Xianlin; Yang, Jingyue; Mancuso, David J; Chen, Jeannie; Bickel, Perry E; Gross, Richard W

    2004-05-01

    Herein, we exploit the power of global lipidomics to identify the critical role of peroxisomal processing of fatty acids in adipocyte lipid storage and metabolism. Remarkably, 3T3-L1 differentiating adipocytes rapidly acquired the ability to alpha oxidize unbranched fatty acids, which is manifested in the accumulation of odd chain length unbranched fatty acids in all major lipid classes. Moreover, in differentiating adipocytes, unsaturated odd chain length fatty acids in TAG molecular species contained exclusively Delta9 olefinic linkages. Unsaturated fatty acids (e.g., oleic and palmitoleic acids) were not subject to alpha oxidation, resulting in the absence of Delta8 unsaturated odd chain length fatty acids. This highly selective substrate utilization resulted in the obligatory sequential ordering of alpha oxidation prior to Delta9 desaturation. On the basis of these results, a putative type 2 peroxisomal localization sequence was identified at the N-terminus of mouse stearoyl-CoA desaturase I (SCD I) comprised of (30)KVKTVPLHL(38). Kinetic analysis demonstrated that the rate of alpha oxidation of exogenously administered [9,10-(3)H]palmitic acid increased 4-fold during differentiation. Similarly, quantitative PCR demonstrated a 4-fold increase in phytanoyl-CoA alpha hydroxylase (PAHX) and fatty acyl-CoA oxidase (FACO) mRNA levels during differentiation. Collectively, these results underscore the role of peroxisomal fatty acid processing as an important determinant of the metabolic fate of fatty acids in the differentiating adipocyte.

  18. Determination of Optimal Parameters for Dual-Layer Cathode of Polymer Electrolyte Fuel Cell Using Computational Intelligence-Aided Design

    PubMed Central

    Chen, Yi; Huang, Weina; Peng, Bei

    2014-01-01

    Because of the demands for sustainable and renewable energy, fuel cells have become increasingly popular, particularly the polymer electrolyte fuel cell (PEFC). Among the various components, the cathode plays a key role in the operation of a PEFC. In this study, a quantitative dual-layer cathode model was proposed for determining the optimal parameters that minimize the over-potential difference and improve the efficiency using a newly developed bat swarm algorithm with a variable population embedded in the computational intelligence-aided design. The simulation results were in agreement with previously reported results, suggesting that the proposed technique has potential applications for automating and optimizing the design of PEFCs. PMID:25490761

  19. Rapid and sensitive determination of tryptophan, serotonin and psychoactive tryptamines by thin-layer chromatography/fluorescence detection.

    PubMed

    Kato, Noriyuki; Kojima, Takashi; Yoshiyagawa, Shinji; Ohta, Hikoto; Toriba, Akira; Nishimura, Hideo; Hayakawa, Kazuichi

    2007-03-23

    A rapid, sensitive and selective method for the determination of tryptophan (Trp), serotonin (5-HT) and psychoactive tryptamines (PATs) by thin-layer chromatography (TLC) with fluorescence detection is proposed. These compounds form fluorophores on the developing plate by heating after spraying with sodium hypochlorite, hydrogen peroxide or potassium hexacyanoferrate(III)-sodium hydroxide reagent. Fluorescent spots (vivid blue) were observed by irradiation with ultraviolet light (365 nm). The detection limits of Trp, 5-HT and PATs were in the range from 0.01 microg to 0.06 microg. This method was effectively applied to the detection of confiscated PAT powder and PAT in abusers' urine samples.

  20. Comparison of lipid deposition at coronary bifurcations versus at nonbifurcation portions of coronary arteries as determined by near-infrared spectroscopy.

    PubMed

    Townsend, Jacob C; Steinberg, Daniel H; Nielsen, Christopher D; Todoran, Thomas M; Patel, Chetan P; Leonardi, Robert A; Wolf, Bethany J; Brilakis, Emmanouil S; Shunk, Kendrick A; Goldstein, James A; Kern, Morton J; Powers, Eric R

    2013-08-01

    Atherosclerosis has been shown to develop preferentially at sites of coronary bifurcation, yet culprit lesions resulting in ST-elevation myocardial infarction do not occur more frequently at these sites. We hypothesized that these findings can be explained by similarities in intracoronary lipid and that lipid and lipid core plaque would be found with similar frequency in coronary bifurcation and nonbifurcation segments. One hundred seventy bifurcations were identified, 156 of which had comparative nonbifurcation segments proximal and/or distal to the bifurcation. We compared lipid deposition at bifurcation and nonbifurcation segments in coronary arteries using near-infrared spectroscopy (NIRS), a novel method for the in vivo detection of coronary lipid. Any NIRS signal for the presence of lipid was found with similar frequency in bifurcation and nonbifurcation segments (79% vs 74%, p = NS). Lipid core burden index, a measure of total lipid quantity indexed to segment length, was similar across bifurcation segments as well as their proximal and distal controls (lipid core burden index 66.3 ± 106, 67.1 ± 116, and 66.6 ± 104, p = NS). Lipid core plaque, identified as a high-intensity focal NIRS signal, was found in 21% of bifurcation segments, and 20% of distal nonbifurcation segments (p = NS). In conclusion, coronary bifurcations do not appear to have higher levels of intracoronary lipid or lipid core plaque than their comparative nonbifurcation regions.

  1. Densitometric thin-layer chromatographic determination of artemisinin and its lipophilic derivatives, artemether and arteether.

    PubMed

    Gabriëls, M; Plaizier-Vercammen, J A

    2003-08-01

    A thin-layer chromatograpy (TLC) method is developed to analyze artemisinin (AT) and its derivatives, artemether (AM) and arteether (AE), using a silica-gel plate with a mobile phase containing pure chloroform. After development, all products are visualized after dipping in a 4-methoxybenzaldehyde dipping reagent of 1% (v/v) in an acidic solution of sulphuric acid (98%, v/v) and acetic acid (96-98%, v/v) (respectively, 2% and 10%, v/v in alcohol-water, 60:30, v/v), presenting a purple color against a slightly colored background. This TLC system is quantitatively evaluated in terms of stability of the color, precision, accuracy, and calibration. Activation is performed at 110 degrees C. Stability of the color of both analytes is reached after 12 min. Precision, less than 5%, is obtained at two levels. Good linearity is obtained in the range of 0.5-8 micro g for all analytes. Some applications show its utility in the quality control of capsules. The prederivatization technique, applying the described dipping reagent before development, reveals the presence of various reaction products, possibly isomers. These results prove that TLC can be a cheap and easy alternative for the analysis of AT and its lipophilic derivatives, AM and AE, as pure powder and in pharmaceutical-dosage forms. PMID:12908952

  2. Semiquantitative determination of polychlorinated biphenyls in tissue samples by thin layer chromatography

    USGS Publications Warehouse

    Mulhern, B.M.; Cromartie, E.; Reichel, W.L.; Belisle, A.A.

    1971-01-01

    A method is described for the analysis of polychlorinated biphenyl (PCB) compounds in tissue samples. Cleanup by hexane-aceto-nitrile partitioning and Florisil column chromatography are performed on samples before oxidative treatment to convert DDE to DCBP. PCB components are then determined semi-quantitatively by TLC. No prior separation of PCB from chlorinated pesticides is required. The lower limit of sensitivity is 0.2 ?g.

  3. Bergenin determination in different extracts by high-performance thin-layer chromatographic densitometry

    PubMed Central

    Khan, Masood Shah; Khan, Washim; Ahmad, Wasim; Singh, Mhaveer; Ahmad, Sayeed

    2015-01-01

    Aim: Bergenin is an active secondary metabolite, found in Bergenia ligulata, family Saxifragaceae, which is an important medicinal plant used in the traditional system of medicine. It is distributed throughout the South and East Asia and some European countries, usually growing on high altitude in the Himalayan region and known as Pashanbhed (meaning “to break the stone”). The rhizome of B. ligulata has been used since long time in different traditional formulations of kidney and liver disorders. Due to its exhaustive use in the traditional system, it is commonly adulterated with the rhizome of other plants which do not contain its chemical marker bergenin. Hence, we developed high-performance thin-layer chromatographic (HPTLC) method for quantification of bergenin in B. ligulata which can be used for its quality control. Materials and Methods: A sensitive HPTLC method has been developed for the estimation of bergenin in different extracts of B. ligulata and its traditional formulations. Precoated HPTLC silica gel plates were used as stationary phase, and chloroform: methanol: acetic acid (8:1:1, v/v/v) was used as mobile phase. Results: The Rf value of bergenin was found to be 0.28 ± 0.03. Detection and quantification were performed by densitometry at 276 nm. The calibration plot was linear in the range of 200–5000 ng of bergenin with the correlation coefficient of (r2) 0.999, which confirms good linearity. The content of bergenin in methanolic and acetone extracts was found to be 5.51 ± 0.14 and 5.76 ± 0.16, respectively. Conclusion: The method can be applied for quality control and standardization of B. ligulata and its traditional formulations as well as for checking the presence of adulterants. PMID:26681881

  4. Comparison of Methods to Determine Tropical Tropopause Layer Cirrus Formation Mechanisms

    SciTech Connect

    Riihimaki, Laura D.; McFarlane, Sally A.; Liang, Calvin; Massie, Steven T.; Beagley, Nathaniel; Toth, Travis D.

    2012-03-30

    A new method of estimating Tropical Tropopause Layer Cirrus (TTLC) formation mechanism (object method) is compared to interpretations of formation from previous studies using back trajectory calculations matched to convection identified from satellites and statistical relationships of TTLC with temperature and water vapor. The object method groups neighboring Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) TTLC cloud profiles into cloud objects and classifies them as convective (35% of TTLC) if they are directly attached to a convective cloud along the CALIPSO track. The percentage of back trajectory calculations that intersect convection (39-95% of TTLC within 5 days) depends strongly on the spatial and temporal resolution of the convection data set, and the manner in which deep convection is identified. Using minimum brightness temperature in 3 hourly, 1{sup o} resolution grid boxes to define convection, 46% of non-convective TTLC (37% of all TTLC) did not trace back to convection within 24 h. Back trajectory calculations of thin cirrus identified by the High Resolution Dynamics Limb Sounder (HIRDLS) gave similar results. Temperature is not a good proxy for formation mechanism as convective and non-convective TTLC frequencies both increase monotonically with decreasing temperature at approximately the same rate. Non-convective TTLC frequencies have a stronger relationship with relative humidity than convective TTLC frequencies, though are not sufficiently different to distinguish object method categories. A decrease in TTL cirrus frequency found at low temperatures in previous studies is caused by insufficient variability in reanalysis temperature data and is not indicative of TTLC formation mechanism.

  5. Contact-free sheet resistance determination of large area graphene layers by an open dielectric loaded microwave cavity

    SciTech Connect

    Shaforost, O.; Wang, K.; Adabi, M.; Guo, Z.; Hanham, S.; Klein, N.; Goniszewski, S.; Gallop, J.; Hao, L.

    2015-01-14

    A method for contact-free determination of the sheet resistance of large-area and arbitrary shaped wafers or sheets coated with graphene and other (semi) conducting ultrathin layers is described, which is based on an open dielectric loaded microwave cavity. The sample under test is exposed to the evanescent resonant field outside the cavity. A comparison with a closed cavity configuration revealed that radiation losses have no significant influence of the experimental results. Moreover, the microwave sheet resistance results show good agreement with the dc conductivity determined by four-probe van der Pauw measurements on a set of CVD samples transferred on quartz. As an example of a practical application, correlations between the sheet resistance and deposition conditions for CVD graphene transferred on quartz wafers are described. Our method has a high potential as measurement standard for contact-free sheet resistance measurement and mapping of large area graphene samples.

  6. MO-G-18C-07: Improving T2 Determination and Quantification of Lipid Methylene Protons in Proton Magnetic Resonance Spectroscopy at 3 T

    SciTech Connect

    Breitkreutz, D.; Fallone, B. G.; Yahya, A.

    2014-06-15

    Purpose: To improve proton magnetic resonance spectroscopy (MRS) transverse relaxation (T{sub 2}) determination and quantification of lipid methylene chain (1.3 ppm) protons by rewinding their J-coupling evolution. Methods: MRS experiments were performed on four lipid phantoms, namely, almond, corn, sunflower and oleic acid, using a 3 T Philips MRI scanner with a transmit/receive birdcage head coil. Two PRESS (Point RESolved Spectroscopy) pulse sequences were used. The first PRESS sequence employed standard bandwidth (BW) (∼550 Hz) RF (radiofrequency) refocussing pulses, while the second used refocussing pulses of narrow BW (∼50 Hz) designed to rewind J-coupling evolution of the methylene protons in the voxel of interest. Signal was acquired with each sequence from a 5×5×5 mm{sup 3} voxel, with a repetition time (TR) of 3000 ms, and with echo times (TE) of 100 to 200 ms in steps of 20 ms. 2048 sample points were measured with a 2000 Hz sampling bandwidth. Additionally, 30 mm outer volume suppression slabs were used to suppress signal outside the voxel of interest. The frequency of the RF pulses was set to that of the methylene resonance. Methylene peak areas were calculated and fitted in MATLAB to a monexponentially decaying function of the form M{sub 0}exp(-TE/T{sub 2}), where M{sub 0} is the extrapolated area when TE = 0 ms and yields a measure of concentration. Results: The determined values of M{sub 0} and T{sub 2} increased for all fatty acids when using the PRESS sequence with narrow BW refocussing pulses. M{sub 0} and T{sub 2} values increased by an average amount (over all the phantoms) of 31% and 14%, respectively. Conclusion: This investigation has demonstrated that J-coupling interactions of lipid methylene protons causes non-negligible signal losses which, if not accounted for, Result in underestimations of their levels and T{sub 2} values when performing MRS measurements. Funded by the Natural Sciences and Engineering Research Council of Canada

  7. Immersion transmission ellipsometry (ITE) for the determination of orientation gradients in photoalignment layers

    NASA Astrophysics Data System (ADS)

    Jung, C. C.; Stumpe, J.

    2014-09-01

    The capability of the method of immersion transmission ellipsometry (ITE) (Jung et al. Int Patent WO, 2004/109260) to not only determine three-dimensional refractive indices in anisotropic thin films (which was already possible in the past), but even their gradients along the z-direction (perpendicular to the film plane) is investigated in this paper. It is shown that the determination of orientation gradients in deep-sub-μm films becomes possible by applying ITE in combination with reflection ellipsometry. The technique is supplemented by atomic force microscopy for measuring the film thickness. For a photo-oriented thin film, no gradient was found, as expected. For a photo-oriented film, which was subsequently annealed in a nematic liquid crystalline phase, an order was found similar to the one applied in vertically aligned nematic displays, with a tilt angle varying along the z-direction. For fresh films, gradients were only detected for the refractive index perpendicular to the film plane, as expected.

  8. Mechanical properties of the superficial biofilm layer determine the architecture of biofilms.

    PubMed

    Kundukad, Binu; Seviour, Thomas; Liang, Yang; Rice, Scott A; Kjelleberg, Staffan; Doyle, Patrick S

    2016-06-29

    Cells in biofilms sense and interact with their environment through the extracellular matrix. The physicochemical properties of the matrix, particularly at the biofilm-environment interface, determine how cells respond to changing conditions. In this study we describe the application of atomic force microscopy and confocal imaging to probe in situ the mechanical properties of these interfacial regions and to elucidate how key matrix components can contribute to the physical sensing by the cells. We describe how the Young's modulus of microcolonies differs according to the size and morphology of microcolonies, as well as the flow rate. The Young's modulus increased as a function of microcolony diameter, which was correlated with the production of the polysaccharide Psl at later stages of maturation for hemispherical or mushroom shaped microcolonies. The Young's modulus of the periphery of the biofilm colony was however independent of the hydrodynamic shear. The morphology of the microcolonies also influenced interfacial or peripheral stiffness. Microcolonies with a diffuse morphology had a lower Young's modulus than isolated, circular ones and this phenomenon was due to a deficiency of Psl. In this way, changes in the specific polysaccharide components imbue the biofilm with distinct physical properties that may modulate the way in which bacteria perceive or respond to their environment. Further, the physical properties of the polysaccharides are closely linked to the specific architectures formed by the developing biofilm. PMID:27273453

  9. Determining concentration fields of tracer plumes for layered porous media in flow-tank experiments

    NASA Astrophysics Data System (ADS)

    Yu, Zhongbo; Schwartz, Franklin W.

    In the laboratory, computer-assisted image analysis provides an accurate and efficient way to monitor tracer experiments. This paper describes the determination of detailed temporal concentration distributions of tracers in a flow-tank experiment by analyzing photographs of plumes of Rhodamine dye through the glass wall of the tank. The methodology developed for this purpose consists of four steps: (1) digitally scanning black and white negatives obtained from photographs of the flow-tank experiment; (2) calibrating and normalizing each digitized image to a standard optical-density scale by determining the relation between the optical density and pixel value for each image; (3) constructing standard curves relating the concentration in an optical density from five experimental runs with predetermined concentrations (2-97mg/L) and (4) converting the optical density to concentration. The spatial distribution of concentration for two photographs was determined by applying these calibration and conversion procedures to all pixels of the digitized images. This approach provides an efficient way to study patterns of plume evolution and transport mechanisms. Résumé Au laboratoire, l'analyse d'images assistée par ordinateur est un moyen précis et efficace pour suivre certaines expériences de traçage. Ce papier présente comment sont déterminées dans le détail les distributions temporelles de la concentration en traceur au cours d'une expérience d'écoulement en réservoir au moyen de l'analyse de photographies de panaches de rhodamine à travers la paroi de verre du réservoir. La méthodologie développée dans cette expérience suit quatre étapes: (1) digitalisation par balayage des négatifs noir et blanc des prises de vue de l'expérience d'écoulement en réservoir (2) calibration et normalisation de chaque image digitalisée par rapport à une échelle étalon de densité optique en déterminant la relation entre la densité optique et la valeur des pixels

  10. Simultaneous determination and pharmacokinetic study of water-soluble and lipid-soluble components of danshen in rat plasma using HPLC-UV method.

    PubMed

    Wang, Xiuli; Zhang, Zhi-Rong; Fu, Hualin; Liu, Jie; Chen, Qian; Nie, Yu; Deng, Li; Gong, Tao

    2007-11-01

    A sensitive and specific HPLC-UV method was developed for the simultaneous determination of major active components of danshen in rat plasma. Both water-soluble and lipid-soluble compounds were included, i.e. danshensu, salvianolic acid B and tanshinone IIA. Protocatechuic aldehyde and diazepam were used as internal standards. The chromatographic separation was achieved on a reversed-phase C(18) column by gradient elution using acetonitrile and 0.025% (v/v) phosphoric acid solution as mobile phase, at a flow rate of 1.0 mL/min. Salvianolic acid B, danshensu and internal standards were detected at 281 nm, while the detection of tanshinone IIA was carried out at 272 nm. All calibration curves showed good linearity (r(2) > 0.999) within test ranges. The limit of detection and the limit of quantification for danshensu, salvianolic acid B and tanshinone IIA in plasma were 0.065, 0.043, 0.022, 0.131, 0.085 and 0.044 microg/mL, respectively. This is the first report on the determination and pharmacokinetic study of danshensu, salvianolic acid B and tanshinone IIA in rat plasma and the results indicated that this method was reliable for the determination of the major active components of danshen in rat plasma. PMID:17685392

  11. An Introduction to Lipid Analysis in the Cell Biology Laboratory.

    ERIC Educational Resources Information Center

    Schuh, Timothy J.

    2002-01-01

    Explains a thin-layer chromatography (TLC) experiment that allows students to study complex mixtures of lipids using small volumes. Uses a water-soluble dye to stain lipids that is fast and safe. (YDS)

  12. Higher smectic-layer order parameters in liquid crystals determined by x-ray diffraction and the effect of antiferroelectricity

    SciTech Connect

    Takanishi, Y.; Ikeda, A.; Takezoe, H.; Fukuda, A. )

    1995-01-01

    By observing the higher-order Bragg peaks corresponding to the layer spacing, the smectic-layer order parameters up to the third order were obtained and the center-of-mass distribution function along the layer normal was calculated. The results clearly indicate that the smectic layer distribution is different from a sinusoidal density wave in antiferroelectric liquid crystals and that the smectic- (Sm) layer order is relatively higher in antiferroelectric Sm-[ital C][sub [ital A

  13. Paternal B Vitamin Intake Is a Determinant of Growth, Hepatic Lipid Metabolism and Intestinal Tumor Volume in Female Apc1638N Mouse Offspring

    PubMed Central

    Sabet, Julia A.; Park, Lara K.; Iyer, Lakshmanan K.; Tai, Albert K.; Koh, Gar Yee; Pfalzer, Anna C.; Parnell, Laurence D.; Mason, Joel B.; Liu, Zhenhua; Byun, Alexander J.; Crott, Jimmy W.

    2016-01-01

    Background The importance of maternal nutrition to offspring health and risk of disease is well established. Emerging evidence suggests paternal diet may affect offspring health as well. Objective In the current study we sought to determine whether modulating pre-conception paternal B vitamin intake alters intestinal tumor formation in offspring. Additionally, we sought to identify potential mechanisms for the observed weight differential among offspring by profiling hepatic gene expression and lipid content. Methods Male Apc1638N mice (prone to intestinal tumor formation) were fed diets containing replete (control, CTRL), mildly deficient (DEF), or supplemental (SUPP) quantities of vitamins B2, B6, B12, and folate for 8 weeks before mating with control-fed wild type females. Wild type offspring were euthanized at weaning and hepatic gene expression profiled. Apc1638N offspring were fed a replete diet and euthanized at 28 weeks of age to assess tumor burden. Results No differences in intestinal tumor incidence or burden were found between male Apc1638N offspring of different paternal diet groups. Although in female Apc1638N offspring there were no differences in tumor incidence or multiplicity, a stepwise increase in tumor volume with increasing paternal B vitamin intake was observed. Interestingly, female offspring of SUPP and DEF fathers had a significantly lower body weight than those of CTRL fed fathers. Moreover, hepatic trigylcerides and cholesterol were elevated 3-fold in adult female offspring of SUPP fathers. Weanling offspring of the same fathers displayed altered expression of several key lipid-metabolism genes. Hundreds of differentially methylated regions were identified in the paternal sperm in response to DEF and SUPP diets. Aside from a few genes including Igf2, there was a striking lack of overlap between these genes differentially methylated in sperm and differentially expressed in offspring. Conclusions In this animal model, modulation of

  14. Analysis of multi-layered films. [determining dye densities by applying a regression analysis to the spectral response of the composite transparency

    NASA Technical Reports Server (NTRS)

    Scarpace, F. L.; Voss, A. W.

    1973-01-01

    Dye densities of multi-layered films are determined by applying a regression analysis to the spectral response of the composite transparency. The amount of dye in each layer is determined by fitting the sum of the individual dye layer densities to the measured dye densities. From this, dye content constants are calculated. Methods of calculating equivalent exposures are discussed. Equivalent exposures are a constant amount of energy over a limited band-width that will give the same dye content constants as the real incident energy. Methods of using these equivalent exposures for analysis of photographic data are presented.

  15. Utilizing SO2/CO2 Mixing Ratios to Determine SO2 Uptake by a Regional Cloud Layer

    NASA Astrophysics Data System (ADS)

    Green, J. R.; Bililign, S.; Fiddler, M. N.; Jaegle, L.; Thornton, J. A.; Brown, S. S.; Shah, V.; Lopez-Hilfiker, F.; Lee, B. H.; Haskins, J.; Fibiger, D. L.; McDuffie, E. E.; Sparks, T.; Ebben, C. J.; Wooldridge, P. J.; Cohen, R. C.; Veres, P. R.; Weinheimer, A. J.; Dibb, J. E.; Schroder, J. C.; Day, D. A.; Jimenez, J. L.; Sullivan, A.; Guo, H.; Weber, R. J.; Campos, T. L.; Leen, J. B.; DiGangi, J. P.; Wolfe, G. M.

    2015-12-01

    The cloud uptake of SO2 during wintertime conditions over the Eastern United States is presented. In wintertime conditions in the Eastern United States there are fewer reactant sinks for gaseous SO2, as a result the atmospheric lifetime is measurably longer. An examination of the SO2/CO2 mixing ratio above and below the cloud layer in comparison to the internal concentration of gaseous CO2 inside the cloud layer will determine uptake of SO2 due to aqueous reactivity SO2/CO2. The performance of this will allow for a numerical estimate of the CO2-H2O complexes compared to the SO2-H2O complexes formed at the cloud droplet surface. The data obtained originates from a series of survey night and day flights on a C-130 aircraft that occurred from Feb 3 to Mar 13, 2015 over the Eastern coastal region of the United States ranging from New York to Florida. The following instruments were utilized in obtaining gaseous measurements; for water droplet size distribution a Condensation Nucleus Counter (CN Counter) (NCAR) and a Forward Scattering Spectrometer Probe, Model 100 (FSSP-100) (NCAR), for SO2 measurements the TECO 43C SO2 analyzer (NOAA—ESRL/NCAT) and a CO/CO2 analyzer (NCAR). An estimate and transport gaseous SO2 due to of cloud uptake in surveyed regions in the WINTER 2015 campaign will be presented.

  16. Locating interfaces in vertically-layered materials and determining concentrations in mixed materials utilizing acoustic impedance measurements

    DOEpatents

    Langlois, G.N.

    1983-09-13

    Measurement of the relative and actual value of acoustic characteristic impedances of an unknown substance, location of the interfaces of vertically-layered materials, and the determination of the concentration of a first material mixed in a second material are disclosed. A highly damped ultrasonic pulse is transmitted into one side of a reference plate, such as a tank wall, where the other side of the reference plate is in physical contact with the medium to be measured. The amplitude of a return signal, which is the reflection of the transmitted pulse from the interface between the other side of the reference plate and the medium, is measured. The amplitude value indicates the acoustic characteristic impedance of the substance relative to that of the reference plate or relative to that of other tested materials. Discontinuities in amplitude with repeated measurements for various heights indicate the location of interfaces in vertically-layered materials. Standardization techniques permit the relative acoustic characteristic impedance of a substance to be converted to an actual value. Calibration techniques for mixtures permit the amplitude to be converted to the concentration of a first material mixed in a second material. 6 figs.

  17. Locating interfaces in vertically-layered materials and determining concentrations in mixed materials utilizing acoustic-impedance measurements. [Patent application

    DOEpatents

    Not Available

    1981-06-10

    Measurement of the relative and actual value of acoustic characteristic impedances of an unknown substance, location of the interfaces of vertically-layered materials, and the determination of the concentration of a first material mixed in a second material are presented. A highly damped ultrasonic pulse is transmitted into one side of a reference plate, such as a tank wall, where the other side of the reference plate is in physical contact with the medium to be measured. The amplitude of a return signal, which is the reflection of the transmitted pulse from the interface between the other side of the reference plate and the medium, is measured. The amplitude value indicates the acoustic characteristic impedance of the substance relative to that of the reference plate or relative to that of other tested materials. Discontinuities in amplitude with repeated measurements for various heights indicate the location of interfaces in vertically-layered materials. Standardization techniques permit the relative acoustic characteristic impedance of a substance to be converted to an actual value. Calibration techniques for mixtures permit the amplitude to be converted to the concentration of a first material mixed in a second material.

  18. Determination of tunnelling parameters in ultra-thin oxide layer poly-Si/SiO 2/Si structures

    NASA Astrophysics Data System (ADS)

    Depas, M.; Vermeire, B.; Mertens, P. W.; Van Meirhaeghe, R. L.; Heyns, M. M.

    1995-08-01

    In this work the electron tunnelling in device grade ultra-thin 3-6 nm n +poly-Si/SiO 2/n-Si structures has been analysed. The well known analytic expression for the Fowler-Nordheim tunnelling current was adapted to include the case of direct tunnelling of electrons, which becomes important for oxide layers thinner than 4.5 nm. For these ultra-thin oxide MOS structures it is necessary to take the band bending in the Si substrate and in the poly-Si layer into account to determine the oxide electrical field strength and to derive the tunnelling parameters of the measured current-voltage characteristic. A method is explained to derive the tunnel barrier height φs and the effective mass of the tunnelling electron mox from the experimental tunnel current characteristics. It is shown that both the direct tunnelling and the Fowler-Nordheim tunnelling current can be quantitatively explained by a WKB approximation using mox as the single fitting parameter.

  19. Large-eddy simulation and stochastic modeling of Lagrangian particles for footprint determination in the stable boundary layer

    NASA Astrophysics Data System (ADS)

    Glazunov, Andrey; Rannik, Üllar; Stepanenko, Victor; Lykosov, Vasily; Auvinen, Mikko; Vesala, Timo; Mammarella, Ivan

    2016-08-01

    Large-eddy simulation (LES) and Lagrangian stochastic modeling of passive particle dispersion were applied to the scalar flux footprint determination in the stable atmospheric boundary layer. The sensitivity of the LES results to the spatial resolution and to the parameterizations of small-scale turbulence was investigated. It was shown that the resolved and partially resolved ("subfilter-scale") eddies are mainly responsible for particle dispersion in LES, implying that substantial improvement may be achieved by using recovering of small-scale velocity fluctuations. In LES with the explicit filtering, this recovering consists of the application of the known inverse filter operator. The footprint functions obtained in LES were compared with the functions calculated with the use of first-order single-particle Lagrangian stochastic models (LSMs) and zeroth-order Lagrangian stochastic models - the random displacement models (RDMs). According to the presented LES, the source area and footprints in the stable boundary layer can be substantially more extended than those predicted by the modern LSMs.

  20. Locating interfaces in vertically-layered materials and determining concentrations in mixed materials utilizing acoustic impedance measurements

    DOEpatents

    Langlois, Gary N.

    1983-09-13

    Measurement of the relative and actual value of acoustic characteristic impedances of an unknown substance, location of the interfaces of vertically-layered materials, and the determination of the concentration of a first material mixed in a second material. A highly damped ultrasonic pulse is transmitted into one side of a reference plate, such as a tank wall, where the other side of the reference plate is in physical contact with the medium to be measured. The amplitude of a return signal, which is the reflection of the transmitted pulse from the interface between the other side of the reference plate and the medium, is measured. The amplitude value indicates the acoustic characteristic impedance of the substance relative to that of the reference plate or relative to that of other tested materials. Discontinuities in amplitude with repeated measurements for various heights indicate the location of interfaces in vertically-layered materials. Standardization techniques permit the relative acoustic characteristic impedance of a substance to be converted to an actual value. Calibration techniques for mixtures permit the amplitude to be converted to the concentration of a first material mixed in a second material.

  1. Determination of the optical band gap of Alq3 and its derivatives for the use in two-layer OLEDs

    NASA Astrophysics Data System (ADS)

    Duvenhage, Mart-Mari; Ntwaeaborwa, Martin; Visser, Hendrik G.; Swarts, Pieter J.; Swarts, Jannie C.; Swart, Hendrik C.

    2015-04-01

    Tris-(8-hydroxyquinolinato)aluminium (Alq3) was used as both the emission and electron transporting layer in organic light emitting diodes. One way to tune the emissive color of Alq3 is to introduce substituents onto the hydroxyquinoline ligands. These substituents will have an effect on the energy gap between the highest occupied molecular orbital and the lowest unoccupied molecular orbital. In this study, Alq3 powders were synthesized with -CH3 substituted at position 5 and 7 (tris-(5,7-dimethyl-8-hydroxyquinolinato)aluminium) and -Cl at position 5 (tris-(5-chloro-8-hydroxyquinolinato) aluminium). Optical absorption and cyclic voltammetry measurements were performed on the samples. The optical band gap was determined from these measurements. The band gap did not vary with more than 0.2 eV from the theoretical value of Alq3.

  2. Employing X-ray Photoelectron Spectroscopy for Determining Layer Homogeneity in Mixed Polar Self-Assembled Monolayers

    PubMed Central

    2016-01-01

    Self-assembled monolayers (SAMs) containing embedded dipolar groups offer the particular advantage of changing the electronic properties of a surface without affecting the SAM–ambient interface. Here we show that such systems can also be used for continuously tuning metal work functions by growing mixed monolayers consisting of molecules with different orientations of the embedded dipolar groups. To avoid injection hot-spots when using the SAM-modified electrodes in devices, a homogeneous mixing of the two components is crucial. We show that a combination of high-resolution X-ray photoelectron spectroscopy with state-of-the-art simulations is an ideal tool for probing the electrostatic homogeneity of the layers and thus for determining phase separation processes in polar adsorbate assemblies down to inhomogeneities at the molecular level. PMID:27429041

  3. Development and validation of a high performance thin layer chromatographic method for determination of 1, 8-Cineole in Callistemon Citrinus

    PubMed Central

    Shaha, Archana; Salunkhe, Vijay R

    2014-01-01

    A new, simple, precise, rapid, and selective high performance thin layer chromatographic (HPTLC) method has been developed and validated for the estimation of 1, 8-cineole in volatile oil of leaves of Callistemon Citrinus obtained by hydro distillation. The method was validated as per ICH guidelines and can be utilized for routine analysis. The retention factor for 1, 8-cineole was found to be 0.52. The linearity was found to be in the range of 3 μg-12 μg. The recovery obtained for 1, 8-cineole was 98%, which is satisfactory. The result obtained in validation indicate the accuracy, reproducibility, and reliability of the developed HPTLC method for determination of 1, 8-cineole. PMID:24761119

  4. Development and validation of a high performance thin layer chromatographic method for determination of 1, 8-Cineole in Callistemon Citrinus.

    PubMed

    Shaha, Archana; Salunkhe, Vijay R

    2014-04-01

    A new, simple, precise, rapid, and selective high performance thin layer chromatographic (HPTLC) method has been developed and validated for the estimation of 1, 8-cineole in volatile oil of leaves of Callistemon Citrinus obtained by hydro distillation. The method was validated as per ICH guidelines and can be utilized for routine analysis. The retention factor for 1, 8-cineole was found to be 0.52. The linearity was found to be in the range of 3 μg-12 μg. The recovery obtained for 1, 8-cineole was 98%, which is satisfactory. The result obtained in validation indicate the accuracy, reproducibility, and reliability of the developed HPTLC method for determination of 1, 8-cineole. PMID:24761119

  5. Development and validation of a high performance thin layer chromatographic method for determination of 1, 8-Cineole in Callistemon Citrinus.

    PubMed

    Shaha, Archana; Salunkhe, Vijay R

    2014-04-01

    A new, simple, precise, rapid, and selective high performance thin layer chromatographic (HPTLC) method has been developed and validated for the estimation of 1, 8-cineole in volatile oil of leaves of Callistemon Citrinus obtained by hydro distillation. The method was validated as per ICH guidelines and can be utilized for routine analysis. The retention factor for 1, 8-cineole was found to be 0.52. The linearity was found to be in the range of 3 μg-12 μg. The recovery obtained for 1, 8-cineole was 98%, which is satisfactory. The result obtained in validation indicate the accuracy, reproducibility, and reliability of the developed HPTLC method for determination of 1, 8-cineole.

  6. In situ Determination of Surface Tension-to-Shear Viscosity Ratio for Quasiliquid Layers on Ice Crystal Surfaces.

    PubMed

    Murata, Ken-Ichiro; Asakawa, Harutoshi; Nagashima, Ken; Furukawa, Yoshinori; Sazaki, Gen

    2015-12-18

    We have experimentally determined the surface tension-to-shear viscosity ratio (the so-called characteristic velocity) of quasiliquid layers (QLLs) on ice crystal surfaces from their wetting dynamics. Using an advanced optical microscope, whose resolution reaches the molecular level in the height direction, we directly observed the coalescent process of QLLs and followed the relaxation modes of their contact lines. The relaxation dynamics is known to be governed by the characteristic velocity, which allows us to access the physical properties of QLLs in a noninvasive way. Here we quantitatively demonstrate that QLLs, when completely wetting ices, have a thickness of 9±3  nm and an approximately 200 times lower characteristic velocity than bulk water, whereas QLLs, when partially wetting ices, have a velocity that is 20 times lower than the bulk. This indicates that ice crystal surfaces significantly affect the physical properties of QLLs localized near the surfaces at a nanometer scale.

  7. Liquid chromatographic determination of aspartame in dry beverage bases and sweetener tablets with confirmation by thin layer chromatography.

    PubMed

    Daniels, D H; Joe, F L; Warner, C R; Fazio, T

    1984-01-01

    A liquid chromatographic method is described for the determination of aspartame in dry beverage bases and sweetener tablets. The sample was mixed with the mobile phase, the pH was adjusted to within +/- 0.1 pH unit of the mobile phase, and the sample was diluted to volume with the mobile phase. The solution was filtered and a 10 microL aliquot was injected onto a C18 reverse phase column. Aspartame was quantitated with an ultraviolet detector. Recoveries of aspartame ranged from 94 to 111%. The dry beverage bases contained 5-13% aspartame and the sweetener tablets contained 19% aspartame. The presence of aspartame was confirmed by using thin layer chromatography.

  8. Employing X-ray Photoelectron Spectroscopy for Determining Layer Homogeneity in Mixed Polar Self-Assembled Monolayers.

    PubMed

    Hehn, Iris; Schuster, Swen; Wächter, Tobias; Abu-Husein, Tarek; Terfort, Andreas; Zharnikov, Michael; Zojer, Egbert

    2016-08-01

    Self-assembled monolayers (SAMs) containing embedded dipolar groups offer the particular advantage of changing the electronic properties of a surface without affecting the SAM-ambient interface. Here we show that such systems can also be used for continuously tuning metal work functions by growing mixed monolayers consisting of molecules with different orientations of the embedded dipolar groups. To avoid injection hot-spots when using the SAM-modified electrodes in devices, a homogeneous mixing of the two components is crucial. We show that a combination of high-resolution X-ray photoelectron spectroscopy with state-of-the-art simulations is an ideal tool for probing the electrostatic homogeneity of the layers and thus for determining phase separation processes in polar adsorbate assemblies down to inhomogeneities at the molecular level. PMID:27429041

  9. Chemical and structural investigation of lipid nanoparticles: drug-lipid interaction and molecular distribution

    NASA Astrophysics Data System (ADS)

    Anantachaisilp, Suranan; Meejoo Smith, Siwaporn; Treetong, Alongkot; Pratontep, Sirapat; Puttipipatkhachorn, Satit; Rungsardthong Ruktanonchai, Uracha

    2010-03-01

    Lipid nanoparticles are a promising alternative to existing carriers in chemical or drug delivery systems. A key challenge is to determine how chemicals are incorporated and distributed inside nanoparticles, which assists in controlling chemical retention and release characteristics. This study reports the chemical and structural investigation of γ-oryzanol loading inside a model lipid nanoparticle drug delivery system composed of cetyl palmitate as solid lipid and Miglyol 812® as liquid lipid. The lipid nanoparticles were prepared by high pressure homogenization at varying liquid lipid content, in comparison with the γ-oryzanol free systems. The size of the lipid nanoparticles, as measured by the photon correlation spectroscopy, was found to decrease with increased liquid lipid content from 200 to 160 nm. High-resolution proton nuclear magnetic resonance (1H-NMR) measurements of the medium chain triglyceride of the liquid lipid has confirmed successful incorporation of the liquid lipid in the lipid nanoparticles. Differential scanning calorimetric and powder x-ray diffraction measurements provide complementary results to the 1H-NMR, whereby the crystallinity of the lipid nanoparticles diminishes with an increase in the liquid lipid content. For the distribution of γ-oryzanol inside the lipid nanoparticles, the 1H-NMR revealed that the chemical shifts of the liquid lipid in γ-oryzanol loaded systems were found at rather higher field than those in γ-oryzanol free systems, suggesting incorporation of γ-oryzanol in the liquid lipid. In addition, the phase-separated structure was observed by atomic force microscopy for lipid nanoparticles with 0% liquid lipid, but not for lipid nanoparticles with 5 and 10% liquid lipid. Raman spectroscopic and mapping measurements further revealed preferential incorporation of γ-oryzanol in the liquid part rather than the solid part of in the lipid nanoparticles. Simple models representing the distribution of γ-oryzanol and

  10. Arsenic bioaccessibility upon gastrointestinal digestion is highly determined by its speciation and lipid-bile salt interactions.

    PubMed

    Alava, Pradeep; Du Laing, Gijs; Odhiambo, Moses; Verliefde, Arne; Tack, Filip; Van de Wiele, Tom R

    2013-01-01

    The release of arsenic (As) from a contaminated food matrix during gastrointestinal digestion, i.e., its bioaccessibility, is an important estimator of its bioavailability, and therefore also its health risk. In addition, As toxicity is primarily determined by its speciation and it is not clear how different As species behave during digestion in the upper digestive tract. Here, we evaluated to what extent digestive parameters like gastric pH and bile concentration, but also food matrix constituents affect the bioaccessibility of 8 As species (As(III), As(V), MMA(V), DMA(V), MMA(III), DMA(III), MMMTA(V), DMMTA(V)). Bioaccessibility of all As standards ranged between 85% and 90% under pH 1.8. Bioaccessibility of methylated and thiolated arsenicals was decreased from 85% to 50% with increasing gastric pH to 4, yet an increasing bile salts concentration up to 30 g/L lowered the bioaccessibility of inorganic species from 83% to 70% due to interaction with Fe present in bile salts. With respect to food matrices, we noted that the fiber content did not affect As bioaccessibility, yet the presence of fat resulted in an increased bioaccessibility of both inorganic and organic arsenicals in the presence of bile salts. With respect to inorganic arsenic, the intestinal presence of trivalent Fe appeared to be the predominant factor for bioaccessibility of iAs. These data demonstrate that species dependent bioaccessibility must be considered upon ingestion and gastrointestinal digestion. PMID:23442116

  11. Lipid nanotechnology.

    PubMed

    Mashaghi, Samaneh; Jadidi, Tayebeh; Koenderink, Gijsje; Mashaghi, Alireza

    2013-01-01

    Nanotechnology is a multidisciplinary field that covers a vast and diverse array of devices and machines derived from engineering, physics, materials science, chemistry and biology. These devices have found applications in biomedical sciences, such as targeted drug delivery, bio-imaging, sensing and diagnosis of pathologies at early stages. In these applications, nano-devices typically interface with the plasma membrane of cells. On the other hand, naturally occurring nanostructures in biology have been a source of inspiration for new nanotechnological designs and hybrid nanostructures made of biological and non-biological, organic and inorganic building blocks. Lipids, with their amphiphilicity, diversity of head and tail chemistry, and antifouling properties that block nonspecific binding to lipid-coated surfaces, provide a powerful toolbox for nanotechnology. This review discusses the progress in the emerging field of lipid nanotechnology. PMID:23429269

  12. Lipid Nanotechnology

    PubMed Central

    Mashaghi, Samaneh; Jadidi, Tayebeh; Koenderink, Gijsje; Mashaghi, Alireza

    2013-01-01

    Nanotechnology is a multidisciplinary field that covers a vast and diverse array of devices and machines derived from engineering, physics, materials science, chemistry and biology. These devices have found applications in biomedical sciences, such as targeted drug delivery, bio-imaging, sensing and diagnosis of pathologies at early stages. In these applications, nano-devices typically interface with the plasma membrane of cells. On the other hand, naturally occurring nanostructures in biology have been a source of inspiration for new nanotechnological designs and hybrid nanostructures made of biological and non-biological, organic and inorganic building blocks. Lipids, with their amphiphilicity, diversity of head and tail chemistry, and antifouling properties that block nonspecific binding to lipid-coated surfaces, provide a powerful toolbox for nanotechnology. This review discusses the progress in the emerging field of lipid nanotechnology. PMID:23429269

  13. Determination of uptake kinetics (sampling rates) by lipid-containing semipermeable membrane devices (SPMDs) for polycyclic aromatic hydrocarbons (PAHs) in water

    USGS Publications Warehouse

    Huckins, J.N.; Petty, J.D.; Orazio, C.E.; Lebo, J.A.; Clark, R.C.; Gibson, V.L.; Gala, W.R.; Echols, K.R.

    1999-01-01

    The use of lipid-containing semipermeable membrane devices (SPMDs) is becoming commonplace, but very little sampling rate data are available for the estimation of ambient contaminant concentrations from analyte levels in exposed SPMDs. We determined the aqueous sampling rates (R(s)s; expressed as effective volumes of water extracted daily) of the standard (commercially available design) 1-g triolein SPMD for 15 of the priority pollutant (PP) polycyclic aromatic hydrocarbons (PAHs) at multiple temperatures and concentrations. Under the experimental conditions of this study, recovery- corrected R(s) values for PP PAHs ranged from ???1.0 to 8.0 L/d. These values would be expected to be influenced by significant changes (relative to this study) in water temperature, degree of biofouling, and current velocity- turbulence. Included in this paper is a discussion of the effects of temperature and octanol-water partition coefficient (K(ow)); the impacts of biofouling and hydrodynamics are reported separately. Overall, SPMDs responded proportionally to aqueous PAH concentrations; i.e., SPMD R(s) values and SPMD-water concentration factors were independent of aqueous concentrations. Temperature effects (10, 18, and 26 ??C) on Rs values appeared to be complex but were relatively small.The use of lipid-containing semipermeable membrane devices (SPMDs) is becoming commonplace, but very little sampling rate data are available for the estimation of ambient contaminant concentrations from analyte levels in exposed SPMDs. We determined the aqueous sampling rates (Rss; expressed as effective volumes of water extracted daily) of the standard (commercially available design) 1-g triolein SPMD for 15 of the priority pollutant (PP) polycyclic aromatic hydrocarbons (PAHs) at multiple temperatures and concentrations. Under the experimental conditions of this study, recovery-corrected Rs values for PP PAHs ranged from ???1.0 to 8.0 L/d. These values would be expected to be influenced by

  14. The Effect of Optic Disc Center Displacement on Retinal Nerve Fiber Layer Measurement Determined by Spectral Domain Optical Coherence Tomography

    PubMed Central

    Uhm, Ki Bang; Sung, Kyung Rim; Kang, Min Ho; Cho, Hee Yoon; Seong, Mincheol

    2016-01-01

    Purpose To investigate the effect of optic disc center displacement on retinal nerve fiber layer (RNFL) measurement determined by spectral domain optical coherence tomography (SD-OCT). Methods The optic disc center was manipulated at 1-pixel intervals in horizontal, vertical, and diagonal directions. According to the manipulated optic disc center location, the RNFL thickness data were resampled: (1) at a 3.46-mm diameter circle; and (2) between a 2.5-mm diameter circle and 5.4-mm square. Error was calculated between the original and resampled RNFL measurements. The tolerable error threshold of the optic disc center displacement was determined by considering test-retest variability of SD-OCT. The unreliable zone was defined as an area with 10% or more variability. Results The maximum tolerable error thresholds of optic disc center displacement on the RNFL thickness map were distributed from 0.042 to 0.09 mm in 8 directions. The threshold shape was vertically elongated. Clinically important unreliable zones were located: (1) at superior and inferior region in the vertical displacement; (2) at inferotemporal region in the horizontal displacement, and (3) at superotemporal or inferotemporal region in the diagonal displacement. The unreliable zone pattern and threshold limit varied according to the direction of optic disc displacement. Conclusions Optic disc center displacement had a considerable impact on whole RNFL thickness measurements. Understanding the effect of optic disc center displacement could contribute to reliable RNFL measurements. PMID:27783663

  15. Determination of neomycin in the form of neomycin derivative with dabsyl chloride by thin layer chromatography and densitometry.

    PubMed

    Hubicka, Urszula; Zuromska-Witek, Barbara; Piotrowska, Joanna; Krzek, Jan

    2015-01-01

    A thin layer chromatographic-densitometric method has been developed for identification and quantitative determination of neomycin derivative with dabsyl chloride. The analysis of antibiotic was achieved on the silica gel TLC plates with fluorescent indicator with n-butanol--2-butanone--25% ammonia--water (10 : 6 : 2 : 2, v/v/v/v) as the mobile phase. The densitometric measurements were made at 460 nm. Under these conditions good separation of chosen aminoglycoside antibiotic from reagent used to make a complex was obtained. The method is characterized by high sensitivity, LOD from 0.1953 μg per band and LOQ from 0.5918 μg per band, wide linearity range from 0.5918 to 2.1960 μg per band for neomycin. The precision of the method was good; RSD varied from 1.17 to 2.05%. Satisfactory results of validation of the method were also confirmed by determination of selected antibiotic in pharmaceutical commercial preparation. The results obtained by TLC-densitometric method were compared with those obtained by spectrophotometric method.

  16. Determination of 2-Aminoacetophenone in wine by high-performance thin-layer chromatography-fluorescence detection.

    PubMed

    Horlacher, Nora; Schwack, Wolfgang

    2016-02-01

    2-Aminoacetophenone (AAP) is closely correlated with the appearance of the sensory phenomenon of UTA ("untypical aging off-flavor") in wine. AAP analyses are generally performed by gas chromatography and mass selective detection (GC/MS), when AAP is extracted from wines by liquid-liquid, solid-liquid or solid phase microextraction. Here we present a rapid, selective and sensitive method for the determination of AAP in wine by high-performance thin-layer chromatography with fluorescence detection (HPTLC-FLD). As internal standard, 2-amino-4-methoxyacetophenone was used. Liquid-liquid extraction with t-butyl methyl ether was followed by a basic cleanup of the extracts, which were applied onto HPTLC amino plates developed with methylene chloride/toluene (7+3, v/v) as mobile phase. Dipping the dried plate into hexane-paraffin solution enhanced fluorescence that was scanned at 366/>400nm. Limits of detection and quantitation were determined to be 0.1 and 0.3μgL(-1) wine, respectively, while only AAP concentrations >0.5μgL(-1) result in UTA. Recoveries were near 100% for model, white, rosé and red wines. Thus, the HPTLC-FLD method enables the analysis of AAP in wines clearly below the odor thresholds and represents a rapid and convenient screening alternative to existing GC/MS methods.

  17. Simultaneous determination of withanolide a and bacoside a in spansules by high-performance thin-layer chromatography.

    PubMed

    Shinde, P B; Aragade, P D; Agrawal, M R; Deokate, U A; Khadabadi, S S

    2011-03-01

    The objective of this work was to develop and validate a simple, rapid, precise, and accurate high performance thin layer chromatography method for simultaneous determination of withanolide A and bacoside A in combined dosage form. The stationary phase used was silica gel G60F(254). The mobile phase used was mixture of ethyl acetate: methanol: toluene: water (4:1:1:0.5 v/v/v/v). The detection of spots was carried out at 320 nm using absorbance reflectance mode. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 200 to 800 ng/spot for withanolide A and 50 to 350 ng/spot for bacoside A. The limit of detection and limit of quantification for the withanolide A were found to be 3.05 and 10.06 ng/spot, respectively and for bacoside A 8.3 and 27.39 ng/spot, respectively. The proposed method can be successfully used to determine the drug content of marketed formulation.

  18. Determination of 2-Aminoacetophenone in wine by high-performance thin-layer chromatography-fluorescence detection.

    PubMed

    Horlacher, Nora; Schwack, Wolfgang

    2016-02-01

    2-Aminoacetophenone (AAP) is closely correlated with the appearance of the sensory phenomenon of UTA ("untypical aging off-flavor") in wine. AAP analyses are generally performed by gas chromatography and mass selective detection (GC/MS), when AAP is extracted from wines by liquid-liquid, solid-liquid or solid phase microextraction. Here we present a rapid, selective and sensitive method for the determination of AAP in wine by high-performance thin-layer chromatography with fluorescence detection (HPTLC-FLD). As internal standard, 2-amino-4-methoxyacetophenone was used. Liquid-liquid extraction with t-butyl methyl ether was followed by a basic cleanup of the extracts, which were applied onto HPTLC amino plates developed with methylene chloride/toluene (7+3, v/v) as mobile phase. Dipping the dried plate into hexane-paraffin solution enhanced fluorescence that was scanned at 366/>400nm. Limits of detection and quantitation were determined to be 0.1 and 0.3μgL(-1) wine, respectively, while only AAP concentrations >0.5μgL(-1) result in UTA. Recoveries were near 100% for model, white, rosé and red wines. Thus, the HPTLC-FLD method enables the analysis of AAP in wines clearly below the odor thresholds and represents a rapid and convenient screening alternative to existing GC/MS methods. PMID:26777086

  19. Quantitative Determination of L-DOPA in Seeds of Mucuna Pruriens Germplasm by High Performance Thin Layer Chromatography.

    PubMed

    Raina, Archana P; Khatri, Renu

    2011-07-01

    Mucuna pruriens Linn. is an important medicinal plant used for treatment of Parkinson's disease and many others in ancient Indian medical system. L-DOPA extracted from seeds of Mucuna is a constituent of more than 200 indigenous drug formulations and is more effective as drug than the synthetic counterpart. A densitometric high performance thin-layer chromatographic (HPTLC) method was developed for quantification of L-DOPA content present in the seeds extract. The method involves separation of L-DOPA on precoated silica gel 60 GF(254) HPTLC plates using a solvent system of n-butanol-acetic-acid-water (4:1:1, v/v) as the mobile phase. Quantification was done at 280 nm using absorbance reflectance mode. Linearity was found in the concentration range of 100 to 1000 ng/spot with the correlation coefficient value of 0.9980. The method was validated for accuracy, precision and repeatability. Mean recovery was 100.89%. The LOD and LOQ for L-DOPA determination were found to be 3.41 ng/spot and 10.35 ng/spot respectively. The proposed HPTLC method was found to be precise, specific and accurate for quantitative determination of L-DOPA. It can be used for rapid screening of large germplasm collections of Mucuna pruriens for L-DOPA content. The method was used to study variation in fifteen accessions of Mucuna germplasm collected from different geographical regions. PMID:22707835

  20. FCS Diffusion Laws in Two-Phase Lipid Membranes: Determination of Domain Mean Size by Experiments and Monte Carlo Simulations

    PubMed Central

    Favard, Cyril; Wenger, Jérôme; Lenne, Pierre-François; Rigneault, Hervé

    2011-01-01

    Many efforts have been undertaken over the last few decades to characterize the diffusion process in model and cellular lipid membranes. One of the techniques developed for this purpose, fluorescence correlation spectroscopy (FCS), has proved to be a very efficient approach, especially if the analysis is extended to measurements on different spatial scales (referred to as FCS diffusion laws). In this work, we examine the relevance of FCS diffusion laws for probing the behavior of a pure lipid and a lipid mixture at temperatures below, within and above the phase transitions, both experimentally and numerically. The accuracy of the microscopic description of the lipid mixtures found here extends previous work to a more complex model in which the geometry is unknown and the molecular motion is driven only by the thermodynamic parameters of the system itself. For multilamellar vesicles of both pure lipid and lipid mixtures, the FCS diffusion laws recorded at different temperatures exhibit large deviations from pure Brownian motion and reveal the existence of nanodomains. The variation of the mean size of these domains with temperature is in perfect correlation with the enthalpy fluctuation. This study highlights the advantages of using FCS diffusion laws in complex lipid systems to describe their temporal and spatial structure. PMID:21354397

  1. Effect of Different Broad Waveband Lights on Membrane Lipids of a Cyanobacterium, Synechococcus sp., as Determined by UPLC-QToF-MS and Vibrational Spectroscopy

    PubMed Central

    Montero, Olimpio; Velasco, Marta; Sanz-Arranz, Aurelio; Rull, Fernando

    2016-01-01

    Differential profile of membrane lipids and pigments of a Synechococcus sp. cyanobacterial strain cells exposed to blue, green, red and white light are determined by means of liquid chromatography and mass spectrometry or diode array detection. Raman and ATR-IR spectra of intact cells under the diverse light wavebands are also reported. Blue light cells exhibited an increased content of photosynthetic pigments as well as specific species of membrane glycerolipids as compared to cells exposed to other wavebands. The A630/A680 ratio indicated an increased content of phycobilisomes (PBS) in the blue light-exposed cells. Some differences in the protein conformation between the four light waveband-exposed cells were deduced from the variable absorbance at specific wavenumbers in the FT-Raman and ATR-FTIR spectra, in particular bands assigned to amide I and amide II. Bands from 1180 to 950 cm−1 in the ATR-FTIR spectrum suggest degraded outer membrane polysaccharide in the blue light-exposed cells. PMID:27223306

  2. Effect of Different Broad Waveband Lights on Membrane Lipids of a Cyanobacterium, Synechococcus sp., as Determined by UPLC-QToF-MS and Vibrational Spectroscopy.

    PubMed

    Montero, Olimpio; Velasco, Marta; Sanz-Arranz, Aurelio; Rull, Fernando

    2016-01-01

    Differential profile of membrane lipids and pigments of a Synechococcus sp. cyanobacterial strain cells exposed to blue, green, red and white light are determined by means of liquid chromatography and mass spectrometry or diode array detection. Raman and ATR-IR spectra of intact cells under the diverse light wavebands are also reported. Blue light cells exhibited an increased content of photosynthetic pigments as well as specific species of membrane glycerolipids as compared to cells exposed to other wavebands. The A630/A680 ratio indicated an increased content of phycobilisomes (PBS) in the blue light-exposed cells. Some differences in the protein conformation between the four light waveband-exposed cells were deduced from the variable absorbance at specific wavenumbers in the FT-Raman and ATR-FTIR spectra, in particular bands assigned to amide I and amide II. Bands from 1180 to 950 cm(-1) in the ATR-FTIR spectrum suggest degraded outer membrane polysaccharide in the blue light-exposed cells. PMID:27223306

  3. Biliary lipid secretion.

    PubMed

    Hişmioğullari, Adnan Adil; Bozdayi, A Mithat; Rahman, Khalid

    2007-06-01

    The liver has many biochemical functions, of which one of the most important is bile formation. Bile is both a secretory and an excretory fluid and two of its most important functions are the delivery to the intestinal tract of: (i) bile acids to assist in fat digestion and absorption; and (ii) liver-derived metabolites of potentially toxic materials prior to their elimination from the body in the feces. Bile contains numerous solutes, including bile acids, phospholipids and cholesterol. Biliary lipids mainly consist of cholesterol and phospholipids and their secretion into bile is affected by the secretion of bile acids. Phospholipids and cholesterol are synthesized in the hepatocytes and are thought to be transferred via vesicle- and non-vesicle-mediated mechanisms into the bile canaliculus. Hepatocytes acquire biliary lipid by three pathways, which are biosynthesis, lipoproteins and existing molecules drawn from intracellular membranes, with the newly synthesized biliary lipid accounting for less than 20% of the total lipids. The hepatic determinants of biliary cholesterol elimination are not limited to total cholesterol homeostasis, but also concern biliary disease conditions, since excess biliary cholesterol secretion is involved in cholesterol gallstone formation, as well as being a major risk factor for gallbladder cancer. The purpose of this review was to highlight some of the major mechanisms involved in biliary lipid secretion.

  4. Lipids and Membrane Lateral Organization

    PubMed Central

    Sonnino, Sandro; Prinetti, Alessandro

    2010-01-01

    Shortly after the elucidation of the very basic structure and properties of cellular membranes, it became evident that cellular membranes are highly organized structures with multiple and multi-dimensional levels of order. Very early observations suggested that the lipid components of biological membranes might be active players in the creation of these levels of order. In the late 1980s, several different and diverse experimental pieces of evidence coalesced together giving rise to the lipid raft hypothesis. Lipid rafts became enormously (and, in the opinion of these authors, sometimes acritically) popular, surprisingly not just within the lipidologist community (who is supposed to be naturally sensitive to the fascination of lipid rafts). Today, a PubMed search using the key word “lipid rafts” returned a list of 3767 papers, including 690 reviews (as a term of comparison, searching over the same time span for a very hot lipid-related key word, “ceramide” returned 6187 hits with 799 reviews), and a tremendous number of different cellular functions have been described as “lipid raft-dependent.” However, a clear consensus definition of lipid raft has been proposed only in recent times, and the basic properties, the ruling forces, and even the existence of lipid rafts in living cells has been recently matter of intense debate. The scenario that is gradually emerging from the controversies elicited by the lipid raft hypothesis emphasizes multiple roles for membrane lipids in determining membrane order, that encompass their tendency to phase separation but are clearly not limited to this. In this review, we would like to re-focus the attention of the readers on the importance of lipids in organizing the fine structure of cellular membranes. PMID:21423393

  5. Conformational flexibility in the apolipoprotein E amino-terminal domain structure determined from three new crystal forms: implications for lipid binding.

    PubMed Central

    Segelke, B. W.; Forstner, M.; Knapp, M.; Trakhanov, S. D.; Parkin, S.; Newhouse, Y. M.; Bellamy, H. D.; Weisgraber, K. H.; Rupp, B.

    2000-01-01

    An amino-terminal fragment of human apolipoprotein E3 (residues 1-165) has been expressed and crystallized in three different crystal forms under similar crystallization conditions. One crystal form has nearly identical cell dimensions to the previously reported orthorhombic (P2(1)2(1)2(1)) crystal form of the amino-terminal 22 kDa fragment of apolipoprotein E (residues 1-191). A second orthorhombic crystal form (P2(1)2(1)2(1) with cell dimensions differing from the first form) and a trigonal (P3(1)21) crystal form were also characterized. The structures of the first orthorhombic and the trigonal form were determined by seleno-methionine multiwavelength anomalous dispersion, and the structure of the second orthorhombic form was determined by molecular replacement using the structure from the trigonal form as a search model. A combination of modern experimental and computational techniques provided high-quality electron-density maps, which revealed new features of the apolipoprotein E structure, including an unambiguously traced loop connecting helices 2 and 3 in the four-helix bundle and a number of multiconformation side chains. The three crystal forms contain a common intermolecular, antiparallel packing arrangement. The electrostatic complimentarity observed in this antiparallel packing resembles the interaction of apolipoprotein E with the monoclonal antibody 2E8 and the low density lipoprotein receptor. Superposition of the model structures from all three crystal forms reveals flexibility and pronounced kinks in helices near one end of the four-helix bundle. This mobility at one end of the molecule provides new insights into the structural changes in apolipoprotein E that occur with lipid association. PMID:10850798

  6. Determining the Origin and Fate of Particulate Plant-Derived Organic Matter in the Rhone River (France) : A Lipid Tracer Review

    NASA Astrophysics Data System (ADS)

    Galeron, M. A.; Amiraux, R.; Charriere, B.; Radakovitch, O.; Raimbault, P.; Garcia, N.; Lagadec, V.; Vaultier, F.; Rontani, J. F.

    2014-12-01

    A number of lipid tracers including fatty acids, hydroxyacids, n-alkanols, sterols and triterpenoids were used to determine the origin and fate of suspended particulate organic matter (POM) collected in the Rhone River (France), with a main focus on phytosterols, such as sitosterol, desmosterol, brassicasterol and cholesterol. This seasonal survey (April 2011 to May 2013) revealed a year-round strong terrigenous contribution to the plant derived particulate organic matter (POM) with significant algal inputs observed in March and attributed to phytoplanktonic blooms likely dominated by diatoms. Specific sitosterol and cholesterol degradation products were quantified and used to estimate the part of biotic and abiotic degradation of POM within the river. Plant-derived organic matter appears to be mainly affected by photo-oxidation and autoxidation (free radical oxidation), while organic matter of human origin, evidenced by the presence of coprostanol, is clearly more prone to bacterial degradation. Despite the involvement of an intense autoxidation inducing homolytic cleavage of peroxy bonds, a significant proportion of hydroperoxides is still intact in higher plant debris. These compounds could play a role in the degradation of terrestrial material by inducing an intense autoxidation upon its arrival at sea. Although sitosterol has been commonly used as a tracer of the terrestrial origin of POM in rivers, we show here that is it also found in phytoplankton, which highlights the need to use different tracers to determine the origin of POM in rivers. As part of the set of tracers we use, we have identified betulin to be an interesting candidate, although limited to a number of angiosperms species. Not only can we trace betulin to an unequivocal terrestrial origin, we also identified its specific degradation products, allowing us to trace the degradation state of angiosperm particulate debris in rivers, as well as the type of degradation undergone.

  7. Lipid synthesis in chick epidermis.

    PubMed

    Lavker, R M

    1975-07-01

    Lipid synthesis in newborn chick epidermis was studied by electron microscopic autoradiography after injection of tritiated palmitate. The labeled lipid product in the tissue was identified as mostly triglyceride. At the earliest time after injection (6 hr), the radioactive precursor was taken up by all viable cells of the epidermis. Grain density was heaviest over basal cells, moderate over spinous cells, and slight over granular cells; thus lipid incorporation is highest in the basal and spinous regions of the chick epidermis. As time after injection progressed, the increasing amounts of grains over the granular and horny cells and decreasing amounts over the basal and spinous cells reflected the continuous upward displacement of cells from one layer into the next. From the distribution of silver grains within the epidermal cells, it has been concluded that, with the passage of time, triglycerides synthesized by the epidermal cells were mainly located in lipid droplets. The numerous grains associated with the elements of the endoplasmic reticulum indicated that this organelle is involved in aggregating triglyceride molecules into lipid droplets. The fact that grains were seen within the horny cells indicated that part of the horny cell consists of lipid probably derived from the lipid droplets retained by the cells during keratinization. PMID:1151110

  8. Lipid Storage Diseases

    MedlinePlus

    ... Awards Enhancing Diversity Find People About NINDS NINDS Lipid Storage Diseases Information Page Condensed from Lipid Storage ... en Español Additional resources from MedlinePlus What are Lipid Storage Diseases? Lipid storage diseases are a group ...

  9. An experimental method to determine the resistance of a vertically aligned carbon nanotube forest in contact with a conductive layer

    NASA Astrophysics Data System (ADS)

    Vo, T. T.; Poulain, C.; Dijon, J.; Fournier, A.; Chevalier, N.; Mariolle, D.

    2012-08-01

    High density vertically aligned carbon nanotube (VACNT) forests are considered as a promising conductive material for many applications (interconnects in microelectronics or contact material layer in sliding contact applications). It is thus crucial to characterize the electrical resistance of these forests, especially in contact with the inherent top/bottom conductive substrates. This paper aims to develop an original method to determine the contribution of the different terms in this electrical resistance, which is measured with a tipless atomic force microscope used in high accuracy "force mode." VACNT stacks with different heights on AlCu substrate with or without Au/Pd top coating are studied. The electrical contact area between the probe tip and the forest is considered to be equivalent to the classical electrical contact area between a tip and a rough surface. With this assumption, the scattering resistance of a mono-wall CNT is 14.6 kΩ μm-1, the top/bottom contact resistance is, respectively, 265 kΩ/385 kΩ. The bottom resistance divided in half is obtained by an interface substrate/CNT catalyst treatment. The same assumption leads to an effective compressive modulus of 175 MPa. These results are consistent with the values published by other authors. The proposed method is effective to optimise the CNT interface contact resistance before integration in a more complex functional structure.

  10. Simultaneous Determination of Size and Quantification of Gold Nanoparticles by Direct Coupling Thin layer Chromatography with Catalyzed Luminol Chemiluminescence

    NASA Astrophysics Data System (ADS)

    Yan, Neng; Zhu, Zhenli; He, Dong; Jin, Lanlan; Zheng, Hongtao; Hu, Shenghong

    2016-04-01

    The increasing use of metal-based nanoparticle products has raised concerns in particular for the aquatic environment and thus the quantification of such nanomaterials released from products should be determined to assess their environmental risks. In this study, a simple, rapid and sensitive method for the determination of size and mass concentration of gold nanoparticles (AuNPs) in aqueous suspension was established by direct coupling of thin layer chromatography (TLC) with catalyzed luminol-H2O2 chemiluminescence (CL) detection. For this purpose, a moving stage was constructed to scan the chemiluminescence signal from TLC separated AuNPs. The proposed TLC-CL method allows the quantification of differently sized AuNPs (13 nm, 41 nm and 100 nm) contained in a mixture. Various experimental parameters affecting the characterization of AuNPs, such as the concentration of H2O2, the concentration and pH of the luminol solution, and the size of the spectrometer aperture were investigated. Under optimal conditions, the detection limits for AuNP size fractions of 13 nm, 41 nm and 100 nm were 38.4 μg L‑1, 35.9 μg L‑1 and 39.6 μg L‑1, with repeatabilities (RSD, n = 7) of 7.3%, 6.9% and 8.1% respectively for 10 mg L‑1 samples. The proposed method was successfully applied to the characterization of AuNP size and concentration in aqueous test samples.

  11. Simultaneous Determination of Size and Quantification of Gold Nanoparticles by Direct Coupling Thin layer Chromatography with Catalyzed Luminol Chemiluminescence.

    PubMed

    Yan, Neng; Zhu, Zhenli; He, Dong; Jin, Lanlan; Zheng, Hongtao; Hu, Shenghong

    2016-01-01

    The increasing use of metal-based nanoparticle products has raised concerns in particular for the aquatic environment and thus the quantification of such nanomaterials released from products should be determined to assess their environmental risks. In this study, a simple, rapid and sensitive method for the determination of size and mass concentration of gold nanoparticles (AuNPs) in aqueous suspension was established by direct coupling of thin layer chromatography (TLC) with catalyzed luminol-H2O2 chemiluminescence (CL) detection. For this purpose, a moving stage was constructed to scan the chemiluminescence signal from TLC separated AuNPs. The proposed TLC-CL method allows the quantification of differently sized AuNPs (13 nm, 41 nm and 100 nm) contained in a mixture. Various experimental parameters affecting the characterization of AuNPs, such as the concentration of H2O2, the concentration and pH of the luminol solution, and the size of the spectrometer aperture were investigated. Under optimal conditions, the detection limits for AuNP size fractions of 13 nm, 41 nm and 100 nm were 38.4 μg L(-1), 35.9 μg L(-1) and 39.6 μg L(-1), with repeatabilities (RSD, n = 7) of 7.3%, 6.9% and 8.1% respectively for 10 mg L(-1) samples. The proposed method was successfully applied to the characterization of AuNP size and concentration in aqueous test samples. PMID:27080702

  12. Simultaneous Determination of Size and Quantification of Gold Nanoparticles by Direct Coupling Thin layer Chromatography with Catalyzed Luminol Chemiluminescence

    PubMed Central

    Yan, Neng; Zhu, Zhenli; He, Dong; Jin, Lanlan; Zheng, Hongtao; Hu, Shenghong

    2016-01-01

    The increasing use of metal-based nanoparticle products has raised concerns in particular for the aquatic environment and thus the quantification of such nanomaterials released from products should be determined to assess their environmental risks. In this study, a simple, rapid and sensitive method for the determination of size and mass concentration of gold nanoparticles (AuNPs) in aqueous suspension was established by direct coupling of thin layer chromatography (TLC) with catalyzed luminol-H2O2 chemiluminescence (CL) detection. For this purpose, a moving stage was constructed to scan the chemiluminescence signal from TLC separated AuNPs. The proposed TLC-CL method allows the quantification of differently sized AuNPs (13 nm, 41 nm and 100 nm) contained in a mixture. Various experimental parameters affecting the characterization of AuNPs, such as the concentration of H2O2, the concentration and pH of the luminol solution, and the size of the spectrometer aperture were investigated. Under optimal conditions, the detection limits for AuNP size fractions of 13 nm, 41 nm and 100 nm were 38.4 μg L−1, 35.9 μg L−1 and 39.6 μg L−1, with repeatabilities (RSD, n = 7) of 7.3%, 6.9% and 8.1% respectively for 10 mg L−1 samples. The proposed method was successfully applied to the characterization of AuNP size and concentration in aqueous test samples. PMID:27080702

  13. Using SANS with Contrast-Matched Lipid Bicontinuous Cubic Phases To Determine the Location of Encapsulated Peptides, Proteins, and Other Biomolecules.

    PubMed

    van 't Hag, Leonie; de Campo, Liliana; Garvey, Christopher J; Feast, George C; Leung, Anna E; Yepuri, Nageshwar Rao; Knott, Robert; Greaves, Tamar L; Tran, Nhiem; Gras, Sally L; Drummond, Calum J; Conn, Charlotte E

    2016-07-21

    An understanding of the location of peptides, proteins, and other biomolecules within the bicontinuous cubic phase is crucial for understanding and evolving biological and biomedical applications of these hybrid biomolecule-lipid materials, including during in meso crystallization and drug delivery. While theoretical modeling has indicated that proteins and additive lipids might phase separate locally and adopt a preferred location in the cubic phase, this has never been experimentally confirmed. We have demonstrated that perfectly contrast-matched cubic phases in D2O can be studied using small-angle neutron scattering by mixing fully deuterated and hydrogenated lipid at an appropriate ratio. The model transmembrane peptide WALP21 showed no preferential location in the membrane of the diamond cubic phase of phytanoyl monoethanolamide and was not incorporated in the gyroid cubic phase. While deuteration had a small effect on the phase behavior of the cubic phase forming lipids, the changes did not significantly affect our results. PMID:27414483

  14. Lipid mobility in supported lipid bilayers by single molecule tracking

    NASA Astrophysics Data System (ADS)

    Kohram, Maryam; Shi, Xiaojun; Smith, Adam

    2015-03-01

    Phospholipid bilayers are the main component of cell membranes and their interaction with biomolecules in their immediate environment is critical for cellular functions. These interactions include the binding of polycationic polymers to lipid bilayers which affects many cell membrane events. As an alternative method of studying live cell membranes, we assemble a supported lipid bilayer and investigate its binding with polycationic polymers in vitro by fluorescently labeling the molecules of the supported lipid bilayer and tracking their mobility. In this work, we use single molecule tracking total internal reflection fluorescence microscopy (TIRF) to study phosphatidylinositol phosphate (PIP) lipids with and without an adsorbed polycationic polymer, quaternized polyvinylpyridine (QPVP). Individual molecular trajectories are obtained from the experiment, and a Brownian diffusion model is used to determine diffusion coefficients through mean square displacements. Our results indicate a smaller diffusion coefficient for the supported lipid bilayers in the presence of QPVP in comparison to its absence, revealing that their binding causes a decrease in lateral mobility.

  15. Iron silicide formation at different layers of (Fe/Si){sub 3} multilayered structures determined by conversion electron Mössbauer spectroscopy

    SciTech Connect

    Badía-Romano, L. Bartolomé, J.; Rubín, J.; Magén, C.; Bürgler, D. E.

    2014-07-14

    The morphology and the quantitative composition of the Fe-Si interface layer forming at each Fe layer of a (Fe/Si){sub 3} multilayer have been determined by means of conversion electron Mössbauer spectroscopy (CEMS) and high-resolution transmission electron microscopy (HRTEM). For the CEMS measurements, each layer was selected by depositing the Mössbauer active {sup 57}Fe isotope with 95% enrichment. Samples with Fe layers of nominal thickness d{sub Fe} = 2.6 nm and Si spacers of d{sub Si} = 1.5 nm were prepared by thermal evaporation onto a GaAs(001) substrate with an intermediate Ag(001) buffer layer. HRTEM images showed that Si layers grow amorphous and the epitaxial growth of the Fe is good only for the first deposited layer. The CEMS spectra show that at all Fe/Si and Si/Fe interfaces a paramagnetic c-Fe{sub 1−x}Si phase is formed, which contains 16% of the nominal Fe deposited in the Fe layer. The bottom Fe layer, which is in contact with the Ag buffer, also contains α-Fe and an Fe{sub 1−x}Si{sub x} alloy that cannot be attributed to a single phase. In contrast, the other two layers only comprise an Fe{sub 1−x}Si{sub x} alloy with a Si concentration of ≃0.15, but no α-Fe.

  16. Hen egg yolk lipid fractions with antiatherogenic properties.

    PubMed

    Nasopoulou, Constantina; Gogaki, Vassiliki; Panagopoulou, Eleanna; Demopoulos, Constantinos; Zabetakis, Ioannis

    2013-03-01

    Three different types of hen egg yolk, cage-free, organic and daily fresh, were tested for their antiatherogenic properties. Total lipids (TL) of all hen egg yolk samples were extracted by the method of Bligh and Dyer and further separated into total polar lipids (TPL) and total neutral lipids (TNL) by counter current distribution chromatography. TPL and TNL were further separated by preparative thin-layer chromatography (TLC). TL, TPL, TNL and the obtained polar and neutral lipid fractions after TLC separation were tested to determine whether they induce platelet activation or inhibit platelet activating factor (PAF)-induced platelet activation. All three hen egg yolk TL samples possessed strong inhibitory activity against PAF-induced platelet activation that was mainly attributed to TPL, especially to PL fraction 4. Cage-free hen egg yolk exhibited the most potent anti-PAF activity in all lipid classes (TL, TPL and TNL). Thus hen egg yolk contains PAF inhibitors that reinforce their nutritional value in terms of protection against cardiovascular diseases, since PAF is a crucial inflammatory phospholipid mediator that is implicated in the mechanism of atherogenesis. PMID:23480708

  17. Covalent attachment of functionalized lipid bilayers to planar waveguides for measuring protein binding to biomimetic membranes.

    PubMed Central

    Heyse, S.; Vogel, H.; Sänger, M.; Sigrist, H.

    1995-01-01

    A new method is presented for measuring sensitively the interactions between ligands and their membrane-bound receptors in situ using integrated optics, thus avoiding the need for additional labels. Phospholipid bilayers were attached covalently to waveguides by a novel protocol, which can in principle be used with any glass-like surface. In a first step, phospholipids carrying head-group thiols were covalently immobilized onto SiO2-TiO2 waveguide surfaces. This was accomplished by acylation of aminated waveguides with the heterobifunctional crosslinker N-succinimidyl-3-maleimidopropionate, followed by the formation of thioethers between the surface-grafted maleimides and the synthetic thiolipids. The surface-attached thiolipids served as hydrophobic templates and anchors for the deposition of a complete lipid bilayer either by fusion of lipid vesicles or by lipid self-assembly from mixed lipid/detergent micelles. The step-by-step lipid bilayer formation on the waveguide surface was monitored in situ by an integrated optics technique, allowing the simultaneous determination of optical thickness and one of the two refractive indices of the adsorbed organic layers. Surface coverages of 50-60% were calculated for thiolipid layers. Subsequent deposition of POPC resulted in an overall lipid layer thickness of 45-50 A, which corresponds to the thickness of a fluid bilayer membrane. Specific recognition reactions occurring at cell membrane surfaces were modeled by the incorporation of lipid-anchored receptor molecules into the supported bilayer membranes. (1) The outer POPC layer was doped with biotinylated phosphatidylethanolamine. Subsequent specific binding of streptavidin was optically monitored. (2) A lipopeptide was incorporated in the outer POPC monolayer. Membrane binding of monoclonal antibodies, which were directed against the peptide moiety of the lipopeptide, was optically detected. The specific antibody binding correlated well with the lipopepitde

  18. Lipid and lipoarabinomannan isolation and characterization.

    PubMed

    Lanéelle, Marie-Antoinette; Nigou, Jérôme; Daffé, Mamadou

    2015-01-01

    Mycobacteria are microorganisms that contain a very high content of structurally diverse lipids, some of them being biologically active substances. As such the lipid composition is commonly used to characterize mycobacterial strains at the species and type-species level. This chapter describes the methods that allow the purification of the most commonly isolated biologically active lipids and those used for analyzing extractable lipids and their constituents, cell wall-linked mycolic acids and lipoarabinomannan (LAM). The latter involve simple chromatographic and analytical techniques, such as thin-layer chromatography and gas chromatography coupled to mass spectrometry.

  19. Ethanol production and maximum cell growth are highly correlated with membrane lipid composition during fermentation as determined by lipidomic analysis of 22 Saccharomyces cerevisiae strains.

    PubMed

    Henderson, Clark M; Lozada-Contreras, Michelle; Jiranek, Vladimir; Longo, Marjorie L; Block, David E

    2013-01-01

    Optimizing ethanol yield during fermentation is important for efficient production of fuel alcohol, as well as wine and other alcoholic beverages. However, increasing ethanol concentrations during fermentation can create problems that result in arrested or sluggish sugar-to-ethanol conversion. The fundamental cellular basis for these problem fermentations, however, is not well understood. Small-scale fermentations were performed in a synthetic grape must using 22 industrial Saccharomyces cerevisiae strains (primarily wine strains) with various degrees of ethanol tolerance to assess the correlation between lipid composition and fermentation kinetic parameters. Lipids were extracted at several fermentation time points representing different growth phases of the yeast to quantitatively analyze phospholipids and ergosterol utilizing atmospheric pressure ionization-mass spectrometry methods. Lipid profiling of individual fermentations indicated that yeast lipid class profiles do not shift dramatically in composition over the course of fermentation. Multivariate statistical analysis of the data was performed using partial least-squares linear regression modeling to correlate lipid composition data with fermentation kinetic data. The results indicate a strong correlation (R(2) = 0.91) between the overall lipid composition and the final ethanol concentration (wt/wt), an indicator of strain ethanol tolerance. One potential component of ethanol tolerance, the maximum yeast cell concentration, was also found to be a strong function of lipid composition (R(2) = 0.97). Specifically, strains unable to complete fermentation were associated with high phosphatidylinositol levels early in fermentation. Yeast strains that achieved the highest cell densities and ethanol concentrations were positively correlated with phosphatidylcholine species similar to those known to decrease the perturbing effects of ethanol in model membrane systems.

  20. Interactions of Lipidic Cubic Phase Nanoparticles with Lipid Membranes.

    PubMed

    Jabłonowska, Elżbieta; Nazaruk, Ewa; Matyszewska, Dorota; Speziale, Chiara; Mezzenga, Raffaele; Landau, Ehud M; Bilewicz, Renata

    2016-09-20

    The interactions of liquid-crystalline monoolein (GMO) cubic phase nanoparticles with various model lipid membranes spread at the air-solution interface by the Langmuir technique were investigated. Cubosomes have attracted attention as potential biocompatible drug delivery systems, and thus understanding their mode of interaction with membranes is of special interest. Cubosomes spreading at the air-water interface as well as interactions with a monolayer of 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) compressed to different surface pressures were studied by monitoring surface pressure-time dependencies at constant area. Progressive incorporation of the nanoparticles was shown to lead to mixed monolayer formation. The concentration of cubosomes influenced the mechanism of incorporation, as well as the fluidity and permeability of the resulting lipid membranes. Brewster angle microscopy images reflected the dependence of the monolayer structure on the cubosomes presence in the subphase. A parameter Csat was introduced to indicate the point of saturation of the lipid membrane with the cubosomal material. This parameter was found to depend on the surface pressure showing that the cubosomes disintegrate in prolonged contact with the membrane, filling available voids in the lipid membrane. At highest surface pressures when the layer is most compact, the penetration of cubosomal material is not possible and only some exchange with the membrane lipid becomes the route of including GMO into the layer. Finally, comparative studies of the interactions between lipids with various headgroup charges with cubosomes suggest that at high surface pressure an exchange of lipid component between the monolayer and the cubosome in its intact form may occur. PMID:27550742

  1. Interactions of Lipidic Cubic Phase Nanoparticles with Lipid Membranes.

    PubMed

    Jabłonowska, Elżbieta; Nazaruk, Ewa; Matyszewska, Dorota; Speziale, Chiara; Mezzenga, Raffaele; Landau, Ehud M; Bilewicz, Renata

    2016-09-20

    The interactions of liquid-crystalline monoolein (GMO) cubic phase nanoparticles with various model lipid membranes spread at the air-solution interface by the Langmuir technique were investigated. Cubosomes have attracted attention as potential biocompatible drug delivery systems, and thus understanding their mode of interaction with membranes is of special interest. Cubosomes spreading at the air-water interface as well as interactions with a monolayer of 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) compressed to different surface pressures were studied by monitoring surface pressure-time dependencies at constant area. Progressive incorporation of the nanoparticles was shown to lead to mixed monolayer formation. The concentration of cubosomes influenced the mechanism of incorporation, as well as the fluidity and permeability of the resulting lipid membranes. Brewster angle microscopy images reflected the dependence of the monolayer structure on the cubosomes presence in the subphase. A parameter Csat was introduced to indicate the point of saturation of the lipid membrane with the cubosomal material. This parameter was found to depend on the surface pressure showing that the cubosomes disintegrate in prolonged contact with the membrane, filling available voids in the lipid membrane. At highest surface pressures when the layer is most compact, the penetration of cubosomal material is not possible and only some exchange with the membrane lipid becomes the route of including GMO into the layer. Finally, comparative studies of the interactions between lipids with various headgroup charges with cubosomes suggest that at high surface pressure an exchange of lipid component between the monolayer and the cubosome in its intact form may occur.

  2. Substrate-supported lipid nanotube arrays.

    SciTech Connect

    Smirnov, A. I.; Poluektov, O. G.; Chemistry; North Carolina State

    2003-07-16

    This Communication describes the self-assembly of phospholipids into lipid nanotubes inside nanoporous anodic aluminum oxide substrate. Orientations of the lipid molecules in such lipid nanoscale structures were verified by high-resolution spin labeling EPR at 95 GHz. The static order parameter of lipids in such nanotube arrays was determined from low-temperature EPR spectra and was found to be exceptionally high, S{sub static} {approx} 0.9. We propose that substrate-supported lipid nanotube arrays have potential for building robust biochips and biosensors in which rigid nanoporous substrates protect the bilayer surface from contamination. The total bilayer surface in the lipid nanotube arrays is much greater than that in the planar substrate-supported membranes. The lipid nanotube arrays seem to be suitable for developing patterned lipid deposition and could be potentially used for patterning of membrane-associated molecules.

  3. A High-Performance Thin Layer Chromatography (HPTLC) Method for Simultaneous Determination of Diphenhydramine Hydrochloride and Naproxen Sodium in Tablets

    PubMed Central

    Bhole, R.P.; Shinde, S.S.; Chitlange, S.S.; Wankhede, S.B.

    2015-01-01

    A rapid and simple high-performance thin layer chromatography (HPTLC) method with densitometry at 230 nm was developed and validated for simultaneous determination of diphenhydramine hydrochloride (DPH) and naproxen sodium (NPS) from pharmaceutical preparation. The separation was carried out on aluminum plates precoated with silica gel 60 F254 using mobile phase toluene:methanol:glacial acetic acid (7.5:1:0.2, v/v/v). The linearity range lies between 200 and 1200 ng/band for DPH and 1760 and 10,560 ng/band for NPS with correlation coefficients of 0.994 and 0.995, respectively. The Rf value for DPH is 0.20 ± 0.05 and for NPS is 0.61 ± 0.06. % Recoveries of DPH and NPS was in the range of 99.70%–99.95% and 99.63%–99.95%, respectively. Limit of detection value for DPH was 13.21 ng/band and for NPS was 8.03 ng/band. Limit of quantitation value for DPH was 40.06 ng/band and for NPS was 24.34 ng/band. The developed method was validated as per ICH guidelines. In stability testing, DPH was found unstable to acid and alkaline hydrolysis, and DPH and NPS were found unstable to oxidation, whereas both the drugs were stable to neutral and photodegradation. The proposed method was successfully applied for the routine quantitative analysis of dosage form containing DPH and NPS. PMID:26692760

  4. Intercalation of small hydrophobic molecules in lipid bilayers containing cholesterol

    SciTech Connect

    Worcester, D.L.; Hamacher, K.; Kaiser, H.; Kulasekere, R.; Torbet, J.

    1994-12-31

    Partitioning of small hydrophobic molecules into lipid bilayers containing cholesterol has been studied using the 2XC diffractometer at the University of Missouri Research Reactor. Locations of the compounds were determined by Fourier difference methods with data from both deuterated and undeuterated compounds introduced into the bilayers from the vapor phase. Data fitting procedures were developed for determining how well the compounds were localized. The compounds were found to be localized in a narrow region at the center of the hydrophobic layer, between the two halves of the bilayer. The structures are therefore intercalated structures with the long axis of the molecules in the plane of the bilayer.

  5. Lipid14: The Amber Lipid Force Field.

    PubMed

    Dickson, Callum J; Madej, Benjamin D; Skjevik, Age A; Betz, Robin M; Teigen, Knut; Gould, Ian R; Walker, Ross C

    2014-02-11

    The AMBER lipid force field has been updated to create Lipid14, allowing tensionless simulation of a number of lipid types with the AMBER MD package. The modular nature of this force field allows numerous combinations of head and tail groups to create different lipid types, enabling the easy insertion of new lipid species. The Lennard-Jones and torsion parameters of both the head and tail groups have been revised and updated partial charges calculated. The force field has been validated by simulating bilayers of six different lipid types for a total of 0.5 μs each without applying a surface tension; with favorable comparison to experiment for properties such as area per lipid, volume per lipid, bilayer thickness, NMR order parameters, scattering data, and lipid lateral diffusion. As the derivation of this force field is consistent with the AMBER development philosophy, Lipid14 is compatible with the AMBER protein, nucleic acid, carbohydrate, and small molecule force fields.

  6. Molecular Interaction of a New Antibacterial Polymer with a Supported Lipid Bilayer Measured by an in situ Label-Free Optical Technique

    PubMed Central

    Horvath, Robert; Kobzi, Balázs; Keul, Helmut; Moeller, Martin; Kiss, Éva

    2013-01-01

    The interaction of the antibacterial polymer–branched poly(ethylene imine) substituted with quaternary ammonium groups, PEO and alkyl chains, PEI25QI5J5A815–with a solid supported lipid bilayer was investigated using surface sensitive optical waveguide spectroscopy. The analysis of the optogeometrical parameters was extended developing a new composite layer model in which the structural and optical anisotropy of the molecular layers was taken into consideration. Following in situ the change of optical birefringence we were able to determine the composition of the lipid/polymer surface layer as well as the displacement of lipid bilayer by the antibacterial polymer without using additional labeling. Comparative assessment of the data of layer thickness and optical anisotropy helps to reveal the molecular mechanism of antibacterial effect of the polymer investigated. PMID:23648479

  7. Molecular interaction of a new antibacterial polymer with a supported lipid bilayer measured by an in situ label-free optical technique.

    PubMed

    Horvath, Robert; Kobzi, Balázs; Keul, Helmut; Moeller, Martin; Kiss, Eva

    2013-01-01

    The interaction of the antibacterial polymer-branched poly(ethylene imine) substituted with quaternary ammonium groups, PEO and alkyl chains, PEI25QI5J5A815-with a solid supported lipid bilayer was investigated using surface sensitive optical waveguide spectroscopy. The analysis of the optogeometrical parameters was extended developing a new composite layer model in which the structural and optical anisotropy of the molecular layers was taken into consideration. Following in situ the change of optical birefringence we were able to determine the composition of the lipid/polymer surface layer as well as the displacement of lipid bilayer by the antibacterial polymer without using additional labeling. Comparative assessment of the data of layer thickness and optical anisotropy helps to reveal the molecular mechanism of antibacterial effect of the polymer investigated. PMID:23648479

  8. Adaptive step-size algorithm for Fourier beam-propagation method with absorbing boundary layer of auto-determined width

    DOE PAGES

    Learn, R.; Feigenbaum, E.

    2016-05-27

    Two algorithms that enhance the utility of the absorbing boundary layer are presented, mainly in the framework of the Fourier beam-propagation method. One is an automated boundary layer width selector that chooses a near-optimal boundary size based on the initial beam shape. Furthermore, the second algorithm adjusts the propagation step sizes based on the beam shape at the beginning of each step in order to reduce aliasing artifacts.

  9. Overexpression of the Epidermis-Specific Homeodomain-Leucine Zipper IV Transcription Factor OUTER CELL LAYER1 in Maize Identifies Target Genes Involved in Lipid Metabolism and Cuticle Biosynthesis1[C][W

    PubMed Central

    Javelle, Marie; Vernoud, Vanessa; Depège-Fargeix, Nathalie; Arnould, Christine; Oursel, Delphine; Domergue, Frédéric; Sarda, Xavier; Rogowsky, Peter M.

    2010-01-01

    Transcription factors of the homeodomain-leucine zipper IV (HD-ZIP IV) family play crucial roles in epidermis-related processes. To gain further insight into the molecular function of OUTER CELL LAYER1 (OCL1), 14 target genes up- or down-regulated in transgenic maize (Zea mays) plants overexpressing OCL1 were identified. The 14 genes all showed partial coexpression with OCL1 in maize organs, and several of them shared preferential expression in the epidermis with OCL1. They encoded proteins involved in lipid metabolism, defense, envelope-related functions, or cuticle biosynthesis and include ZmWBC11a (for white brown complex 11a), an ortholog of AtWBC11 involved in the transport of wax and cutin molecules. In support of the annotations, OCL1-overexpressing plants showed quantitative and qualitative changes of cuticular wax compounds in comparison with wild-type plants. An increase in C24 to C28 alcohols was correlated with the transcriptional up-regulation of ZmFAR1, coding for a fatty acyl-coenzyme A reductase. Transcriptional activation of ZmWBC11a by OCL1 was likely direct, since transactivation in transiently transformed maize kernels was abolished by a deletion of the activation domain in OCL1 or mutations in the L1 box, a cis-element bound by HD-ZIP IV transcription factors. Our data demonstrate that, in addition to AP2/EREBP and MYB-type transcription factors, members of the HD-ZIP IV family contribute to the transcriptional regulation of genes involved in cuticle biosynthesis. PMID:20605912

  10. LIPIDS OF SARCINA LUTEA I.

    PubMed Central

    Huston, Charles K.; Albro, Phillip W.

    1964-01-01

    Huston, Charles K. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.), and Phillip W. Albro. Lipids of Sarcina lutea. I. Fatty acid composition of the extractable lipids. J. Bacteriol. 88:425–432. 1964.—The extractable lipids of Sarcina lutea were separated into several fractions by a combination of column and thin-layer chromatography. Qualitative and quantitative characterization of the fatty acid content of these lipid fractions was accomplished by means of gas-liquid chromatography and infrared analyses. Of the total extract, the lipids consisted of 2.1% free fatty acids, 51.0% glycerides, and 22.7% complex lipids; they had a fatty acid content with a complete spectrum of carbon numbers from C8 to C22. The fatty acids included a large component of branched-acids in addition to the normal straight-chain acids. The branched-acids, comprising 40% of the fatty acids analyzed, constituted a homologous series of iso-acids from C12 to C19. Two 18-carbon unsaturates were found cis-9-octadecenoate and cis-11-octadecenoate. A relatively high percentage (20.5%) of the extractable material from S. lutea was found to be hydrocarbon. This material was not further characterized. PMID:14203360

  11. Two Layer Model for Local Tear Film Dynamics

    NASA Astrophysics Data System (ADS)

    Gewecke, Nicholas; Braun, Rich; Breward, Chris; King-Smith, Ewen

    2012-11-01

    Many tear film models utilize a single-layer approach that represents only the aqueous layer, which constitutes the majority of the tear film. In such models, the layer is dominated by shear stresses. Some recent models have incorporated surfactant effects at the liquid-air interface to model the effects of polar lipids there. Clinical observations of the lipid layer indicate more complicated dynamics of the lipid layer than demonstrated by these previous models. The model presented in this talk includes a thin lipid layer between the aqueous layer and the air, which is treated as an extensional flow. Our results demonstrate formation of lipid drops, with the number of drops dependent upon the parameters of the system, especially the thickness ratio between the lipid and aqueous layers.

  12. Isolation and chemical characterization of lipid A from gram-negative bacteria.

    PubMed

    Henderson, Jeremy C; O'Brien, John P; Brodbelt, Jennifer S; Trent, M Stephen

    2013-09-16

    Lipopolysaccharide (LPS) is the major cell surface molecule of gram-negative bacteria, deposited on the outer leaflet of the outer membrane bilayer. LPS can be subdivided into three domains: the distal O-polysaccharide, a core oligosaccharide, and the lipid A domain consisting of a lipid A molecular species and 3-deoxy-D-manno-oct-2-ulosonic acid residues (Kdo). The lipid A domain is the only component essential for bacterial cell survival. Following its synthesis, lipid A is chemically modified in response to environmental stresses such as pH or temperature, to promote resistance to antibiotic compounds, and to evade recognition by mediators of the host innate immune response. The following protocol details the small- and large-scale isolation of lipid A from gram-negative bacteria. Isolated material is then chemically characterized by thin layer chromatography (TLC) or mass-spectrometry (MS). In addition to matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS, we also describe tandem MS protocols for analyzing lipid A molecular species using electrospray ionization (ESI) coupled to collision induced dissociation (CID) and newly employed ultraviolet photodissociation (UVPD) methods. Our MS protocols allow for unequivocal determination of chemical structure, paramount to characterization of lipid A molecules that contain unique or novel chemical modifications. We also describe the radioisotopic labeling, and subsequent isolation, of lipid A from bacterial cells for analysis by TLC. Relative to MS-based protocols, TLC provides a more economical and rapid characterization method, but cannot be used to unambiguously assign lipid A chemical structures without the use of standards of known chemical structure. Over the last two decades isolation and characterization of lipid A has led to numerous exciting discoveries that have improved our understanding of the physiology of gram-negative bacteria, mechanisms of antibiotic resistance, the human innate

  13. An Empirical Approach to Determining the Boundary Layer Bromine Monoxide (BrO) Abundance from Satellite Total Column Measurements

    NASA Astrophysics Data System (ADS)

    Simpson, W. R.; Donohoue, D.; Carlson, D. A.

    2009-12-01

    Unique chemistry in the Arctic boundary layer during springtime liberates bromine from sea salt, producing reactive halogen gases (e.g. atomic bromine and bromine monoxide radicals) that then drastically alter atmospheric oxidation pathways. This phenomenon causes ozone depletion events and affects mercury deposition to the snowpack. Satellite remote sensing (e.g. OMI and GOME2 observations) techniques can detect the total column abundance of BrO, which is generally the primary species of the reactive bromine family. However, BrO is also present in the stratosphere, so it is necessary to partition the satellite-observed BrO column abundance into boundary layer and non-boundary layer (primarily stratospheric) partial columns to be able to infer boundary layer abundances and hence chemical affects near the Earth's surface (i.e. ozone and mercury impacts). In this presentation, we describe an empirical method for partitioning the BrO total column and apply it globally during spring 2008. The method indicates that some BrO total column enhancements ("hotspots") are not actually enhancements in the boundary layer BrO abundance but occur aloft. Movies and a statistical analysis of the inferred boundary layer BrO abundance are presented. The method has been tested and performs well at the Barrow field site. However, in areas that lack routine ground truth BrO measurements (e.g. Hudson Bay, Canada), large tropospheric BrO abundances are indicated. It is not clear if these inferred boundary layer BrO events are real or if the simple empirical method described here is failing in those locations. Verification of this method over large spatial regions of the Arctic is needed.

  14. Membrane lipid alterations in hemoglobinopathies.

    PubMed

    Kuypers, Frans A

    2007-01-01

    The red blood cell (RBC) membrane is a complex mixture of lipids and proteins. Hundreds of phospholipid molecular species spontaneously arrange themselves in a lipid bilayer and move rapidly in the plane as well as across the bilayer in a dynamic but highly organized fashion. Areas enriched in certain lipids determine proper protein function. Phospholipids are asymmetrically distributed across the lipid bilayer with phosphatidylserine (PS) exclusively on the inside. Both the composition and organization of the RBC membrane is well maintained. Alterations lead to apoptosis during erythropoiesis or early demise of the cell in the circulation. The mechanisms that govern the maintenance of the lipid bilayer are only recently being unraveled at the individual protein level. Oxidized lipids are rapidly repaired using fatty acids taken up from plasma to maintain membrane integrity. Several isoforms of a RBC acyl-Coenzyme A (CoA) synthase have been reported, as well as the first member of a family of lysophospholipid acylCoA acyltransferases. Phospholipid asymmetry is maintained by the recently identified RBC amino-phospholipid translocase. These enzymes, essential in maintaining membrane lipid organization, are affected by oxidant stress or an increase in cytosolic calcium. Normal lipid composition and organization is lost in subpopulations of RBC in hemoglobinopathies such as sickle cell disease and thalassemia. Despite elaborate antioxidant systems, lipids and membrane proteins, including those that maintain lipid organization, are damaged in these cells. This in turn leads to improper repair of damaged RBC membranes and altered interactions of RBCs with other blood cells and plasma components that play a role in the pathology that defines these disorders. The altered lipid bilayer in RBCs in hemoglobinopathies leads to premature removal (anemia) and imbalance in hemostasis, and plays a role in vaso-occlusive crisis in sickle cell disease. Lipid breakdown products of PS

  15. Determination of the emission zone in a single-layer polymer light-emitting diode through optical measurements

    SciTech Connect

    Granlund, Thomas; Pettersson, Leif A. A.; Inganas, Olle

    2001-06-01

    We study the emission zone in a single-layer polymer light-emitting diode. The emission zone is found by studying the angular distribution of the electroluminescence. The emission is modeled by accounting for optical interference. We account for birefringence of the anode layer in our model. The active polymer was, however, found to be isotropic. The anode consists of a single-layer of the conducting polymer complex poly(3,4-ethylenedioxythiophene) and poly(styrene sulfonate) (PEDOT-PSS), with enhanced conductivity. As a cathode we use plain aluminum. By using only PEDOT-PSS we avoid having a thin metal layer or indium-tin-oxide as the anode in the path of the escaping light. The active material is a substituted polythiophene with excellent film forming properties. A comparison between the experimental and calculated angular distribution of light emission from a single-layered polymer light-emitting diode was shown to be in good agreement for the spectral region studied. By assuming a distribution of the emission zone, we deduce the position as well as the width of the zone. {copyright} 2001 American Institute of Physics.

  16. Determination of Boundary-Layer Transition on Three Symmetrical Airfoils in the NACA Full-Scale Wind Tunnel

    NASA Technical Reports Server (NTRS)

    Silverstein, Abe; Becker, John V

    1938-01-01

    For the purpose of studying the transition from laminar to turbulent flow, boundary-layer measurements were made in the NACA full-scale wind tunnel on three symmetrical airfoils of NACA 0009, 0012, and 0018 sections. The effects of variations in lift coefficient, Reynolds number, and airfoil thickness on transition were investigated. Air speed in the boundary layer was measured by total-head tubes and by hot wires; a comparison of transition as indicated by the two techniques was obtained. The results indicate no unique value of Reynolds number for the transition, whether the Reynolds number is based upon the distance along the chord or upon the thickness of the boundary layer at the transition point. In general, the transition is not abrupt and occurs in a region that varies in length as a function of the test conditions.

  17. Structure and dynamics of the lipid modifications of a transmembrane α-helical peptide determined by ²H solid-state NMR spectroscopy.

    PubMed

    Penk, Anja; Müller, Matthias; Scheidt, Holger A; Langosch, Dieter; Huster, Daniel

    2011-03-01

    The fusion of biological membranes is mediated by integral membrane proteins with α-helical transmembrane segments. Additionally, those proteins are often modified by the covalent attachment of hydrocarbon chains. Previously, a series of de novo designed α-helical peptides with mixed Leu/Val sequences was presented, mimicking fusiogenically active transmembrane segments in model membranes (Hofmann et al., Proc. Natl. Acad. Sci. USA 101 (2004) 14776-14781). From this series, we have investigated the peptide LV16 (KKKW LVLV LVLV LVLV LVLV KKK), which was synthesized featuring either a free N-terminus or a saturated N-acylation of 2, 8, 12, or 16 carbons. We used ²H and ³¹P NMR spectroscopy to investigate the structure and dynamics of those peptide lipid modifications in POPC and DLPC bilayers and compared them to the hydrocarbon chains of the surrounding membrane. Except for the C2 chain, all peptide acyl chains were found to insert well into the membrane. This can be explained by the high local lipid concentrations the N-terminal lipid chains experience. Further, the insertion of these peptides did not influence the membrane structure and dynamics as seen from the ²H and ³¹P NMR data. In spite of the fact that the longer acyl chains insert into the membrane, they do not adapt their lengths to the thickness of the bilayer. Even the C16 lipid chain on the peptide, which could match the length of the POPC palmitoyl chain, exhibited lower order parameters in the upper chain, which get closer and finally reach similar values in the lower chain region. ²H NMR square law plots reveal motions of slightly larger amplitudes for the peptide lipid chains compared to the surrounding phospholipids. In spite of the significantly different chain lengths of the acylations, the fraction of gauche defects in the inserted chains is constant.

  18. Stearoyl-CoA Desaturase 1 Is a Key Determinant of Membrane Lipid Composition in 3T3-L1 Adipocytes

    PubMed Central

    Hagen, Rachel; Vidal-Puig, Antonio

    2016-01-01

    Stearoyl-CoA desaturase 1 (SCD1) is a lipogenic enzyme important for the regulation of membrane lipid homeostasis; dysregulation likely contributes to obesity associated metabolic disturbances. SCD1 catalyses the Δ9 desaturation of 12-19 carbon saturated fatty acids to monounsaturated fatty acids. To understand its influence in cellular lipid composition we investigated the effect of genetic ablation of SCD1 in 3T3-L1 adipocytes on membrane microdomain lipid composition at the species-specific level. Using liquid chromatography/electrospray ionisation-tandem mass spectrometry, we quantified 70 species of ceramide, mono-, di- and trihexosylceramide, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, bis(monoacylglycero)phosphate, phosphatidylinositol and cholesterol in 3T3-L1 adipocytes in which a 90% reduction in scd1 mRNA expression was achieved with siRNA. Cholesterol content was unchanged although decreases in other lipids resulted in cholesterol accounting for a higher proportion of lipid in the membranes. This was associated with decreased membrane lateral diffusion. An increased ratio of 24:0 to 24:1 in ceramide, mono- and dihexosylceramide, and sphingomyelin likely also contributed to this decrease in lateral diffusion. Of particular interest, we observed a decrease in phospholipids containing arachidonic acid. Given the high degree of structural flexibility of this acyl chain this will influence membrane lateral diffusion, and is likely responsible for the transcriptional activation of Lands’ cycle enzymes lpcat3 and mboat7. Of relevance these profound changes in the lipidome were not accompanied by dramatic changes in gene expression in mature differentiated adipocytes, suggesting that adaptive homeostatic mechanisms to ensure partial maintenance of the biophysical properties of membranes likely occur at a post-transcriptional level. PMID:27632198

  19. Stearoyl-CoA Desaturase 1 Is a Key Determinant of Membrane Lipid Composition in 3T3-L1 Adipocytes.

    PubMed

    Rodriguez-Cuenca, Sergio; Whyte, Lauren; Hagen, Rachel; Vidal-Puig, Antonio; Fuller, Maria

    2016-01-01

    Stearoyl-CoA desaturase 1 (SCD1) is a lipogenic enzyme important for the regulation of membrane lipid homeostasis; dysregulation likely contributes to obesity associated metabolic disturbances. SCD1 catalyses the Δ9 desaturation of 12-19 carbon saturated fatty acids to monounsaturated fatty acids. To understand its influence in cellular lipid composition we investigated the effect of genetic ablation of SCD1 in 3T3-L1 adipocytes on membrane microdomain lipid composition at the species-specific level. Using liquid chromatography/electrospray ionisation-tandem mass spectrometry, we quantified 70 species of ceramide, mono-, di- and trihexosylceramide, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, bis(monoacylglycero)phosphate, phosphatidylinositol and cholesterol in 3T3-L1 adipocytes in which a 90% reduction in scd1 mRNA expression was achieved with siRNA. Cholesterol content was unchanged although decreases in other lipids resulted in cholesterol accounting for a higher proportion of lipid in the membranes. This was associated with decreased membrane lateral diffusion. An increased ratio of 24:0 to 24:1 in ceramide, mono- and dihexosylceramide, and sphingomyelin likely also contributed to this decrease in lateral diffusion. Of particular interest, we observed a decrease in phospholipids containing arachidonic acid. Given the high degree of structural flexibility of this acyl chain this will influence membrane lateral diffusion, and is likely responsible for the transcriptional activation of Lands' cycle enzymes lpcat3 and mboat7. Of relevance these profound changes in the lipidome were not accompanied by dramatic changes in gene expression in mature differentiated adipocytes, suggesting that adaptive homeostatic mechanisms to ensure partial maintenance of the biophysical properties of membranes likely occur at a post-transcriptional level. PMID:27632198

  20. Structural studies of polymer-cushioned lipid bilayers.

    PubMed

    Majewski, J; Wong, J Y; Park, C K; Seitz, M; Israelachvili, J N; Smith, G S

    1998-11-01

    The structure of softly supported polymer-cushioned lipid bilayers, prepared in two different ways at the quartz-solution interface, were determined using neutron reflectometry. The polymer cushion consisted of a thin layer of branched, cationic polyethyleneimine (PEI), and the bilayers were formed by adsorption of small unilamellar dimyristoylphosphatidylcholine (DMPC) vesicles. When vesicles were first allowed to adsorb to a bare quartz substrate, an almost perfect bilayer formed. When the polymer was then added to the aqueous solution, it appeared to diffuse beneath this bilayer, effectively lifting it from the substrate. In contrast, if the polymer layer is adsorbed first to the bare quartz substrate followed by addition of vesicles to the solution, there is very little interaction of the vesicles with the polymer layer, and the result is a complex structure most likely consisting of patchy multilayers or adsorbed vesicles.

  1. The physics of stratum corneum lipid membranes.

    PubMed

    Das, Chinmay; Olmsted, Peter D

    2016-07-28

    The stratum corneum (SC), the outermost layer of skin, comprises rigid corneocytes (keratin-filled dead cells) in a specialized lipid matrix. The continuous lipid matrix provides the main barrier against uncontrolled water loss and invasion of external pathogens. Unlike all other biological lipid membranes (such as intracellular organelles and plasma membranes), molecules in the SC lipid matrix show small hydrophilic groups and large variability in the length of the alkyl tails and in the numbers and positions of groups that are capable of forming hydrogen bonds. Molecular simulations provide a route for systematically probing the effects of each of these differences separately. In this article, we present the results from atomistic molecular dynamics of selected lipid bilayers and multi-layers to probe the effect of these polydispersities. We address the nature of the tail packing in the gel-like phase, the hydrogen bond network among head groups, the bending moduli expected for leaflets comprising SC lipids and the conformation of very long ceramide lipids in multi-bilayer lipid assemblies.This article is part of the themed issue 'Soft interfacial materials: from fundamentals to formulation'.

  2. Composition determination of quaternary GaAsPN layers from single X-ray diffraction measurement of quasi-forbidden (002) reflection

    SciTech Connect

    Tilli, J.-M. Jussila, H.; Huhtio, T.; Sopanen, M.; Yu, K. M.

    2014-05-28

    GaAsPN layers with a thickness of 30 nm were grown on GaP substrates with metalorganic vapor phase epitaxy to study the feasibility of a single X-ray diffraction (XRD) measurement for full composition determination of quaternary layer material. The method is based on the peak intensity of a quasi-forbidden (002) reflection, which is shown to vary with changing arsenic content for GaAsPN. The method works for thin films with a wide range of arsenic contents and shows a clear variation in the reflection intensity as a function of changing layer composition. The obtained thicknesses and compositions of the grown layers are compared with accurate reference values obtained by Rutherford backscattering spectroscopy combined with nuclear reaction analysis measurements. Based on the comparison, the error in the XRD defined material composition becomes larger with increasing nitrogen content and layer thickness. This suggests that the dominating error source is the deteriorated crystal quality due to the nonsubstitutional incorporation of nitrogen into the crystal lattice and strain relaxation. The results reveal that the method overestimates the arsenic and nitrogen content within error margins of about 0.12 and about 0.025, respectively.

  3. Experimental determination of the boundary layer at air-sample inlet positions on the NASA CV 990 aircraft

    NASA Technical Reports Server (NTRS)

    Bowen, S. W.; Vedder, J. F.; Condon, E. P.

    1984-01-01

    Full-scale, in-flight measurements of the boundary-layer thickness, velocity profile, and flow angle have been made at several sample collection stations on the fuselage of the NASA CV 990. These results are given as functions of Mach number, Reynolds number, yaw, and angle of attack.

  4. A robust post-processing method to determine skin friction in turbulent boundary layers from the velocity profile

    NASA Astrophysics Data System (ADS)

    Rodríguez-López, Eduardo; Bruce, Paul J. K.; Buxton, Oliver R. H.

    2015-04-01

    The present paper describes a method to extrapolate the mean wall shear stress, , and the accurate relative position of a velocity probe with respect to the wall, , from an experimentally measured mean velocity profile in a turbulent boundary layer. Validation is made between experimental and direct numerical simulation data of turbulent boundary layer flows with independent measurement of the shear stress. The set of parameters which minimize the residual error with respect to the canonical description of the boundary layer profile is taken as the solution. Several methods are compared, testing different descriptions of the canonical mean velocity profile (with and without overshoot over the logarithmic law) and different definitions of the residual function of the optimization. The von Kármán constant is used as a parameter of the fitting process in order to avoid any hypothesis regarding its value that may be affected by different initial or boundary conditions of the flow. Results show that the best method provides an accuracy of for the estimation of the friction velocity and for the position of the wall. The robustness of the method is tested including unconverged near-wall measurements, pressure gradient, and reduced number of points; the importance of the location of the first point is also tested, and it is shown that the method presents a high robustness even in highly distorted flows, keeping the aforementioned accuracies if one acquires at least one data point in . The wake component and the thickness of the boundary layer are also simultaneously extrapolated from the mean velocity profile. This results in the first study, to the knowledge of the authors, where a five-parameter fitting is carried out without any assumption on the von Kármán constant and the limits of the logarithmic layer further from its existence.

  5. Stacking sequence determines Raman intensities of observed interlayer shear modes in 2D layered materials--A general bond polarizability model.

    PubMed

    Luo, Xin; Lu, Xin; Cong, Chunxiao; Yu, Ting; Xiong, Qihua; Quek, Su Ying

    2015-01-01

    2D layered materials have recently attracted tremendous interest due to their fascinating properties and potential applications. The interlayer interactions are much weaker than the intralayer bonds, allowing the as-synthesized materials to exhibit different stacking sequences, leading to different physical properties. Here, we show that regardless of the space group of the 2D materials, the Raman frequencies of the interlayer shear modes observed under the typical z(xx)z configuration blue shift for AB stacked materials, and red shift for ABC stacked materials, as the number of layers increases. Our predictions are made using an intuitive bond polarizability model which shows that stacking sequence plays a key role in determining which interlayer shear modes lead to the largest change in polarizability (Raman intensity); the modes with the largest Raman intensity determining the frequency trends. We present direct evidence for these conclusions by studying the Raman modes in few layer graphene, MoS2, MoSe2, WSe2 and Bi2Se3, using both first principles calculations and Raman spectroscopy. This study sheds light on the influence of stacking sequence on the Raman intensities of intrinsic interlayer modes in 2D layered materials in general, and leads to a practical way of identifying the stacking sequence in these materials. PMID:26469313

  6. Combined optical and acoustical method for determination of thickness and porosity of transparent organic layers below the ultra-thin film limit

    NASA Astrophysics Data System (ADS)

    Rodenhausen, K. B.; Kasputis, T.; Pannier, A. K.; Gerasimov, J. Y.; Lai, R. Y.; Solinsky, M.; Tiwald, T. E.; Wang, H.; Sarkar, A.; Hofmann, T.; Ianno, N.; Schubert, M.

    2011-10-01

    Analysis techniques are needed to determine the quantity and structure of materials composing an organic layer that is below an ultra-thin film limit and in a liquid environment. Neither optical nor acoustical techniques can independently distinguish between thickness and porosity of ultra-thin films due to parameter correlation. A combined optical and acoustical approach yields sufficient information to determine both thickness and porosity. We describe application of the combinatorial approach to measure single or multiple organic layers when the total layer thickness is small compared to the wavelength of the probing light. The instrumental setup allows for simultaneous in situ spectroscopic ellipsometry and quartz crystal microbalance dynamic measurements, and it is combined with a multiple-inlet fluid control system for different liquid solutions to be introduced during experiments. A virtual separation approach is implemented into our analysis scheme, differentiated by whether or not the organic adsorbate and liquid ambient densities are equal. The analysis scheme requires that the film be assumed transparent and rigid (non-viscoelastic). We present and discuss applications of our approach to studies of organic surfactant adsorption, self-assembled monolayer chemisorption, and multiple-layer target DNA sensor preparation and performance testing.

  7. Combined optical and acoustical method for determination of thickness and porosity of transparent organic layers below the ultra-thin film limit.

    PubMed

    Rodenhausen, K B; Kasputis, T; Pannier, A K; Gerasimov, J Y; Lai, R Y; Solinsky, M; Tiwald, T E; Wang, H; Sarkar, A; Hofmann, T; Ianno, N; Schubert, M

    2011-10-01

    Analysis techniques are needed to determine the quantity and structure of materials composing an organic layer that is below an ultra-thin film limit and in a liquid environment. Neither optical nor acoustical techniques can independently distinguish between thickness and porosity of ultra-thin films due to parameter correlation. A combined optical and acoustical approach yields sufficient information to determine both thickness and porosity. We describe application of the combinatorial approach to measure single or multiple organic layers when the total layer thickness is small compared to the wavelength of the probing light. The instrumental setup allows for simultaneous in situ spectroscopic ellipsometry and quartz crystal microbalance dynamic measurements, and it is combined with a multiple-inlet fluid control system for different liquid solutions to be introduced during experiments. A virtual separation approach is implemented into our analysis scheme, differentiated by whether or not the organic adsorbate and liquid ambient densities are equal. The analysis scheme requires that the film be assumed transparent and rigid (non-viscoelastic). We present and discuss applications of our approach to studies of organic surfactant adsorption, self-assembled monolayer chemisorption, and multiple-layer target DNA sensor preparation and performance testing. PMID:22047284

  8. Milk lipid secretion: recent biomolecular aspects

    PubMed Central

    McManaman, James L.

    2014-01-01

    Neonates of most species depend on milk lipids for calories, fat-soluble vitamins, and bioactive lipid components for growth and development during the postnatal period. To meet neonatal nutrition and development needs, the mammary gland has evolved efficient mechanisms for synthesizing and secreting large quantities of lipid during lactation. Although the biochemical steps involved in milk lipid synthesis are understood, the identities of the genes mediating these steps and the molecular physiology of milk lipid production and secretion have only recently begun to be understood in detail through advances in mouse genetics, gene expression analysis, protein structural properties, and the cell biology of lipid metabolism. This review discusses emerging data about the molecular, cellular, and structural determinants of milk lipid synthesis and secretion within the context of physiological functions. PMID:24605173

  9. Lipid dynamics at dendritic spines.

    PubMed

    Dotti, Carlos Gerardo; Esteban, Jose Antonio; Ledesma, María Dolores

    2014-01-01

    Dynamic changes in the structure and composition of the membrane protrusions forming dendritic spines underlie memory and learning processes. In recent years a great effort has been made to characterize in detail the protein machinery that controls spine plasticity. However, we know much less about the involvement of lipids, despite being major membrane components and structure determinants. Moreover, protein complexes that regulate spine plasticity depend on specific interactions with membrane lipids for proper function and accurate intracellular signaling. In this review we gather information available on the lipid composition at dendritic spine membranes and on its dynamics. We pay particular attention to the influence that spine lipid dynamism has on glutamate receptors, which are key regulators of synaptic plasticity.

  10. A mechanism of raft formation on both plasma membrane layers

    NASA Astrophysics Data System (ADS)

    Sornbundit, Kan; Modchang, Charin; Triampo, Wannapong; Triampo, Darapond; Nuttavut, Narin

    2013-10-01

    A double-layered membrane model is proposed to explain raft formation and induction on extracellular (outer) and cytoplasmic (inner) leaflets of plasma membranes in a situation where only the outer layer has a tendency to phase-separate. In the model, lipid exchange with the surrounding medium is allowed on both layers, but lipid exchange between layers is not allowed. Simulations display domain stabilization on both layers. The effect of the lipid recycling frequencies on stationary domain sizes is also investigated. It is found that stationary domain sizes decrease when lipid recycling frequencies are stronger. Linear stability analysis is used to verify the results.

  11. Evaluation of lipid removal from animal fats for the determination of organochlorine, pesticides, and polychlorinated biphenyl indicators by gas chromatography with electron capture detector.

    PubMed

    Andrade, Adriana S; Sacheto, Doriane; Hoff, Rodrigo B; Pizzolato, Tania M

    2012-09-01

    For cleanup of animal fat before GC analysis of organochlorine pesticides and polychlorinated biphenyls, freezing-lipid filtration with solid-phase extraction and matrix solid-phase dispersion were evaluated to replace the official Brazilian methodology that uses preparative alumina column chromatography. General drawbacks associated with this last technique, such as the use of large amounts of solvent, laborious and time-consuming procedure could be avoided by using these alternative approaches. Experiments were carried out to study the performance by using different combinations of sorbents and elution solvents. Efficiency of alternative extraction methods in terms of fat removal and recovery capability was monitored by gravimetry, TLC, and GC with electron capture detection. Freezing-lipid filtration with solid-phase extraction afforded better clean up efficiency with recoveries in a range of 54.5 to 103.6% with the relative standard deviation of less than 10% for all compounds under investigation.

  12. Chain length specificity for activation of cPLA2alpha by C1P: use of the dodecane delivery system to determine lipid-specific effects.

    PubMed

    Wijesinghe, Dayanjan S; Subramanian, Preeti; Lamour, Nadia F; Gentile, Luciana B; Granado, Maria H; Bielawska, Alicja; Szulc, Zdzislaw; Gomez-Munoz, Antonio; Chalfant, Charles E

    2009-10-01

    Previously, our laboratory demonstrated that ceramide-1-phosphate (C1P) specifically activated group IVA cytosolic phospholipase A(2) (cPLA(2)alpha) in vitro. In this study, we investigated the chain length specificity of this interaction. C1P with an acyl-chain of >or=6 carbons efficiently activated cPLA(2)alpha in vitro, whereas C(2)-C1P, was unable to do so. Delivery of C1P to cells via the newly characterized ethanol/dodecane system demonstrated a lipid-specific activation of cPLA(2)alpha, AA release, and PGE(2) synthesis (EC(50) = 400 nM) when compared to structurally similar lipids. C1P delivered as vesicles in water also induced a lipid-specific increase in AA release. Mass spectrometric analysis demonstrated that C1P delivered via ethanol/dodecane induced a 3-fold increase in endogenous C1P with little metabolism to ceramide. C1P was also more efficiently delivered (>3-fold) to internal membranes by ethanol/dodecane as compared to vesiculated C1P. Using this now established delivery method for lipids, C(2)-C1P was shown to be ineffective in the induction of AA release as compared with C(6)-C1P, C(16)-C1P, and C(18:1) C1P. Here, we demonstrate that C1P requires >or=6 carbon acyl-chain to activate cPLA(2)alpha. Thus, published reports on the biological activity of C(2)-C1P are not via eicosanoid synthesis. Furthermore, this study demonstrates that the alcohol/dodecane system can be used to efficiently deliver exogenous phospholipids to cells for the examination of specific biological effects.

  13. Determination of aflatoxins in air samples of refuse-derived fuel by thin-layer chromatography with laser-induced fluorescence spectrometric detection

    SciTech Connect

    Bicking, M.K.L.; Kniseley, R.N.; Svec, H.J.

    1983-02-01

    An analytical method is described which allows determination of aflatoxins in a complex matrix. An apparatus has been developed that quantitates fluorescent compounds on thin-layer chromatography plates. A nitrogen laser excitation source produces a detection limit of 10 pg for four aflatoxins. Aflatoxin B1 has been found at levels up to 17 ppb in solid samples collected from the air at a plant which produces refuse-derived fuel. 7 figures, 1 table.

  14. Local environment and composition of magnesium gallium layered double hydroxides determined from solid-state 1H and 71Ga NMR spectroscopy

    SciTech Connect

    Petersen, Line B.; Lipton, Andrew S.; Zorin, Vadim; Nielsen, Ulla Gro

    2014-11-01

    Ordering of gallium(III) in a series of magnesium gallium layered double hydroxides (LDH’s), [Mg1-xGax(OH)2(NO3)x yH2O], was determined using solid-state 1H and 71Ga NMR spectroscopy. Depletion of Ga in these LDH’s is demonstrated to be the result of soluble [Ga(OH)4]-complexes formed during synthesis.

  15. Diversity of Lipid Distribution in Fish Skeletal Muscle.

    PubMed

    Kaneko, Gen; Shirakami, Hirohito; Hirano, Yuki; Oba, Moemi; Yoshinaga, Hazuki; Khieokhajonkhet, Anurak; Nagasaka, Reiko; Kondo, Hidehiro; Hirono, Ikuo; Ushio, Hideki

    2016-04-01

    Adipose tissue is a lipid storage organ characterized by the pronounced accumulation of adipocytes. Although adipose tissues are found in various parts of the vertebrate body, it is unclear whether these tissues have a common ancestral origin or have evolved in several phylogenetic lineages by independent adipocyte accumulation events. To gain insight into the evolutionary history of vertebrate adipose tissues, we determined the distribution of adipocytes by oil red O staining in skeletal muscle of 10 teleost species spanning eight orders: Tetraodontiformes, Pleuronectiformes, Spariformes, Salmoniformes, Clupeiformes, Beloniformes, Osmeriformes, and Cypriniformes. Accumulation of adipocytes in the myoseptum was observed in many species, including red seabream, rainbow trout, Pacific herring, Pacific saury, zebrafish and giant danio. We also found some order-, species-, and swimming mode-specific distribution patterns of adipocytes: 1) almost complete absence of intramuscular adipocytes in the order Tetraodontiformes (torafugu and spotted green pufferfish), 2) clear adipocyte accumulation in the inclinator muscles of fin in Japanese flounder, 3) a large intramuscular adipose tissue at the root of the dorsal fin in ayu, and 4) thick lipid layers consisting of subcutaneous adipose tissue and red muscle lipids in pelagic migratory fish (Pacific herring and Pacific saury). Of note, Pacific herring and Pacific saury are phylogenetically distinct species sharing a similar niche and swimming mode, suggesting that their analogous adipocyte/lipid distribution patterns are the consequence of convergent evolution. The potentially heterogeneous origin of adipose tissues has significant implications for the interpretation of their functional diversity. PMID:27032682

  16. Determination of preservatives in cosmetic products. I. Thin-layer chromatographic procedure for the identification of preservatives in cosmetic products.

    PubMed

    De Kruijf, N; Rijk, M A; Pranoto-Soetardhi, L A; Schouten, A

    1987-12-11

    A thin-layer chromatographic procedure is presented for the separation and identification of preservatives that are listed in the current EEC Council Directive on cosmetic products or have been permitted in the past. The method consists of an extraction of acidified cosmetics with methanol, separation of the extracts by thin-layer chromatography on aluminium oxide and silica gel-coated plates using one developing solvent, and visualization of the preservatives on the plates using short-wavelength UV light and six detection reagents. The retention behaviour and the detectability of 88 preservatives were investigated, of which 74 were characterized by this method. The preservatives in fourteen commercial cosmetic products were tentatively identified by the procedure described. In general this method will permit the routine detection of preservatives in cosmetics in an approximate concentration of 0.1% (w/w).

  17. Synthesis, characterization and structure determination of two isotypes of a layered aluminophosphate with a new 2D network topology

    SciTech Connect

    Tuel, A. . E-mail: tuel@catalyse.cnrs.fr; Lorentz, Ch.; Gramlich, V.; Baerlocher, Ch.

    2005-07-15

    Two isotypes of a new layered aluminophosphate, further denoted MDAP-3 and MDAE-1, have been synthesized under hydrothermal conditions using N-methyl-1,3-propanediamine and N-methyl-ethylenediamine, respectively. MDAP-3, with the empirical formula [Al{sub 2}(HPO{sub 4})(PO{sub 4}){sub 2}](C{sub 4}N{sub 2}H{sub 14})(H{sub 2}O), crystallizes in the orthorhombic space group Pna2(1) (No. 33) with a=9.602(16)A, b=9.26(2)A, c=16.03(3)A, Z=4, R{sub 1}=0.0498 and wR{sub 2}=0.1217. The second solid, MDAE-1, with the empirical formula [Al{sub 2}(HPO{sub 4})(PO{sub 4}){sub 2}](C{sub 3}N{sub 2}H{sub 12})(H{sub 2}O), crystallizes in the same space group with a=9.4250(19)A, b=9.3170(19)A, c=15.907(3)A, Z=4, R{sub 1}=0.0407 and wR{sub 2}=0.0954. The two compounds possess the same layer topology. Inorganic layers contain PO{sub 3}=O, PO{sub 3}OH, AlO{sub 4} and AlO{sub 6} polyhedra, linked together to generate a new 4x8 net. MDAP-3 and MDAE-1 represent the first examples of two-dimensional layered aluminophosphates with the Al{sub 2}P{sub 3}O{sub 12} stoichiometry, and containing AlO{sub 6} octahedra.

  18. Pattern of inner retinal layers involvement in pigmented paravenous retinochoroidal atrophy as determined by SD-OCT: case report.

    PubMed

    Junqueira, Daniela Laura Melo; Lopes, Flavio Siqueira Santos; Biteli, Luís Gustavo; Prata, Tiago Santos

    2013-01-01

    Pigmented paravenous retinochoroidal atrophy is an ocular disease characterized by outer retina and choroidal atrophy often with overlying intraretinal bone spicule pigment deposition along the retinal veins. As a rare condition, there is scant information in the literature regarding the pattern of inner retinal layers involvement. We present a case of a 41-year-old white man initially referred for a glaucoma evaluation. Fundoscopy revealed patches of retinochoroidal atrophy and light pigmentation extending from the optic nerve head along the inferior-temporal retinal veins in both eyes. Using different spectral-domain optical coherence tomography (SD-OCT) protocols we identified a significant thinning of the inner retinal layers along the inferior-temporal veins, but with a lucid interval surrounding the optic nerve head. Standard automated perimetry revealed a superior absolute arcuate scotoma sparing the central fixation (good structure-functional correlation). This pattern of inner retinal layers involvement was not previously described. We believe SD-OCT added significantly to the anatomical description of this case. Physicians should consider these new anatomical findings and correlate them with functional status while assessing these patients.

  19. Determination of the critical product layer thickness in the reaction of CaO with CO{sub 2}

    SciTech Connect

    Alvarez, D.; Abanades, J.C.

    2005-07-20

    Calcium oxide can be an effective CO{sub 2} sorbent at high temperatures. When coupled with a calcination step to produce pure CO{sub 2}, the carbonation reaction is the basis for several high-temperature separation systems of CO{sub 2}. The formation of a product layer of CaCO{sub 3} is known to mark a sudden change in the reaction regime, from a very fast CO{sub 2} uptake to very slow carbonation rates. The critical thickness of this product layer of CaCO{sub 3} has been measured in this work on real sorbent materials, using different limestone precursors and submitting them to many repeated carbonation calcination cycles (up to 100). Mercury porosimetry curves of the calcines and their carbonated counterparts have been obtained and their differences interpreted with a simple pore model, from which the thickness of the product layer is derived. An average value of 49 nm (19% standard deviation) has been obtained, which is quite insensitive to the type of limestone and to the texture of the calcine as long as the model is fulfilled. The implications of this value on our understanding of the sorbent performance in these CO{sub 2}-capture systems are discussed.

  20. Influence of alloy inhomogeneities on the determination by Raman scattering of composition and strain in Si1-xGex/Si(001) layers

    NASA Astrophysics Data System (ADS)

    Reparaz, J. S.; Marcus, I. C.; Goñi, A. R.; Garriga, M.; Alonso, M. I.

    2012-07-01

    In this work, we investigate the influence of alloy composition inhomogeneities on the vibrational properties of strained Si1-xGex/Si layers with x ranging from 0 to 0.5. We show that the frequencies of the principal alloy vibrational modes (Ge-Ge, Si-Ge, and Si-Si) are strongly influenced by the distribution of Ge atoms within the alloy layers, which becomes gradually random following a series of sequential annealing steps. Our measurements suggest that the composition dependence of the optical phonon frequencies in fully random and unstrained alloys is well described by the results previously published by Alonso and Winer [Phys. Rev. B 39, 10056 (1989)]. In the general case of an alloy layer with unknown degree of compositional inhomogeneity and/or strain relaxation, though the analysis of the Raman spectra is not straightforward. Therefore, we propose an analytical/graphical method to accurately estimate the Ge content and residual strain of SiGe layers exhibiting any level of compositional disorder or strain status, by performing a single Raman measurement. This would be extremely useful in situations where x-ray measurements cannot be conducted. We show that our procedure to treat the Raman data holds for the whole compositional range but with different accuracy depending upon the case: (i) For annealed SiGe layers (mostly strain relaxed) the Ge content x can be directly determined with high accuracy of ±0.01. (ii) For strained samples (usually as-grown samples) an extra criterion must be adopted seeking for a graphical solution, accounting for the degree of compositional inhomogeneity. In this case, the error in the determination of Ge content depends on alloy composition, being the upper bound ±0.02 for x <0.3 and ±0.03 for x >0.3.

  1. Painted supported lipid membranes

    PubMed Central

    Florin, E.-L.; Gaub, H. E.

    1993-01-01

    We report herein measurements on a novel type of supported lipid films, which we call painted supported membranes (PSM). These membranes are formed in a self-assembly process on alkylated gold films from an organic solution. The formation process was investigated with surface plasmon resonance microscopy. The optical and electrical properties of the films were determined for various types of lipids and as a function of temperature by means of cyclic voltammetry and potential relaxation after charge injection. We could show that these films exhibit an extraordinarily high specific resistivity which, depending on the lipid, may be as high as 109 ohm/cm2. We could also show that due to this low conductivity, an electrical polarization across the PSM relaxes with characteristic time constants of up to 20 min. The electrical properties together with their high mechanical stability and accessibility to surface sensitive techniques make these films well suitable model membranes for optical and electrical investigations. Examples for such applications are given in the subsequent article by Seifert et al. ImagesFIGURE 3FIGURE 4 PMID:19431873

  2. Self-assembly formation of lipid bilayer coatings on bare aluminum oxide: overcoming the force of interfacial water.

    PubMed

    Jackman, Joshua A; Tabaei, Seyed R; Zhao, Zhilei; Yorulmaz, Saziye; Cho, Nam-Joon

    2015-01-14

    Widely used in catalysis and biosensing applications, aluminum oxide has become popular for surface functionalization with biological macromolecules, including lipid bilayer coatings. However, it is difficult to form supported lipid bilayers on aluminum oxide, and current methods require covalent surface modification, which masks the interfacial properties of aluminum oxide, and/or complex fabrication techniques with specific conditions. Herein, we addressed this issue by identifying simple and robust strategies to form fluidic lipid bilayers on aluminum oxide. The fabrication of a single lipid bilayer coating was achieved by two methods, vesicle fusion under acidic conditions and solvent-assisted lipid bilayer (SALB) formation under near-physiological pH conditions. Importantly, quartz crystal microbalance with dissipation (QCM-D) monitoring measurements determined that the hydration layer of a supported lipid bilayer on aluminum oxide is appreciably thicker than that of a bilayer on silicon oxide. Fluorescence recovery after photobleaching (FRAP) analysis indicated that the diffusion coefficient of lateral lipid mobility was up to 3-fold greater on silicon oxide than on aluminum oxide. In spite of this hydrodynamic coupling, the diffusion coefficient on aluminum oxide, but not silicon oxide, was sensitive to the ionic strength condition. Extended-DLVO model calculations estimated the thermodynamics of lipid-substrate interactions on aluminum oxide and silicon oxide, and predict that the range of the repulsive hydration force is greater on aluminum oxide, which in turn leads to an increased equilibrium separation distance. Hence, while a strong hydration force likely contributes to the difficulty of bilayer fabrication on aluminum oxide, it also confers advantages by stabilizing lipid bilayers with thicker hydration layers due to confined interfacial water. Such knowledge provides the basis for improved surface functionalization strategies on aluminum oxide

  3. Self-assembly formation of lipid bilayer coatings on bare aluminum oxide: overcoming the force of interfacial water.

    PubMed

    Jackman, Joshua A; Tabaei, Seyed R; Zhao, Zhilei; Yorulmaz, Saziye; Cho, Nam-Joon

    2015-01-14

    Widely used in catalysis and biosensing applications, aluminum oxide has become popular for surface functionalization with biological macromolecules, including lipid bilayer coatings. However, it is difficult to form supported lipid bilayers on aluminum oxide, and current methods require covalent surface modification, which masks the interfacial properties of aluminum oxide, and/or complex fabrication techniques with specific conditions. Herein, we addressed this issue by identifying simple and robust strategies to form fluidic lipid bilayers on aluminum oxide. The fabrication of a single lipid bilayer coating was achieved by two methods, vesicle fusion under acidic conditions and solvent-assisted lipid bilayer (SALB) formation under near-physiological pH conditions. Importantly, quartz crystal microbalance with dissipation (QCM-D) monitoring measurements determined that the hydration layer of a supported lipid bilayer on aluminum oxide is appreciably thicker than that of a bilayer on silicon oxide. Fluorescence recovery after photobleaching (FRAP) analysis indicated that the diffusion coefficient of lateral lipid mobility was up to 3-fold greater on silicon oxide than on aluminum oxide. In spite of this hydrodynamic coupling, the diffusion coefficient on aluminum oxide, but not silicon oxide, was sensitive to the ionic strength condition. Extended-DLVO model calculations estimated the thermodynamics of lipid-substrate interactions on aluminum oxide and silicon oxide, and predict that the range of the repulsive hydration force is greater on aluminum oxide, which in turn leads to an increased equilibrium separation distance. Hence, while a strong hydration force likely contributes to the difficulty of bilayer fabrication on aluminum oxide, it also confers advantages by stabilizing lipid bilayers with thicker hydration layers due to confined interfacial water. Such knowledge provides the basis for improved surface functionalization strategies on aluminum oxide

  4. Comparison of sulfuric acid treatment and multi-layer silica gel column chromatography in cleanup methods for determination of PCDDs, PCDFs and dioxin-like PCBs in foods.

    PubMed

    Amakura, Yoshiaki; Tsutsumi, Tomoaki; Sasaki, Kumiko; Toyoda, Masatake; Maitani, Tamio

    2002-10-01

    Two typical cleanup methods, sulfuric acid treatment and multi-layer silica gel column chromatography, for the determination of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) in seventeen food samples were examined and compared. Vegetables, fruits, cereals, fish, meat and dairy foods were extracted by conventional methods (shaking with acetone/n-hexane or with n-hexane after alkaline treatment). The extracts were cleaned up by sulfuric acid treatment or multi-layer silica gel column chromatography, followed by several column chromatographic steps. Of the samples treated, the vegetable, fruit and cereal samples could be directly applied to the multi-layer silica gel column after extraction. However, the samples containing fats and oils such as fish, meat and dairy foods needed to be treated several times with concentrated sulfuric acid before multi-layer column chromatography, because these samples plugged the column with oily residues. Both cleanup methods gave similar values of isomeric concentrations and showed similar efficiency of purification, and the recoveries ranged from 40 to 120%. These results are considered to provide useful data for the efficient analysis of dioxins in foods which have wide-ranging compositions.

  5. Determination of Planetary Boundary Layer Heights on Short Spatial and Temporal Scales from Surface and Airborne Vertical Profilers during DISCOVER-AQ

    NASA Astrophysics Data System (ADS)

    Delgado, R.; Berkoff, T.; Compton, J. S.; St Pé, A.; Baker, B.; Hoff, R. M.; Martins, D. K.; Thompson, A. M.; Yang, E.; Christopher, S. A.; Joseph, E.; Tzortziou, M.; Lolli, S.; Landry, L.; Woodman, M.; Ferrare, R. A.; Hostetler, C. A.

    2011-12-01

    The National Research Council [2009] identified lower troposphere profiling of trace gases, aerosol and thermodynamic quantities as a cross-cutting need for air quality, weather, climate, energy and other national priority economic areas. A critical parameter in determining air pollution concentrations near the ground is the depth through which pollutants are vigorously mixed. The planetary boundary layer height (PBLH) is an important meteorological parameter that affects near-surface atmospheric pollutant concentrations since it determines the volume of air into which pollutants and their precursors are emitted. This height is also important in determining the relationship between column measures of gases and aerosols and the concentration measured at the surface, since pollutants are frequently contained within the PBL. Determining the mixing in the PBL was one goal of a study of the spatial and diurnal variations of the PBLH over Maryland for July 2011, during NASA's Earth Venture mission DISCOVER-AQ. The PBLHs were obtained from elastic lidars (surface and airborne) and wind profiler observations by determining the convective mixed layer using the covariance wavelet technique (CWT) and comparing it to the virtual potential temperature measurements from soundings. This July, a total of 16 ozone episode days (National Ambient Air Quality Standard 8 hour ozone concentrations greater than 75 ppb) occurred. The relationship between PBLH and surface ozone concentrations was evaluated against ventilation coefficients (product of the PBLH times the surface wind speed) and PBL growth rates during ozone episode and non-episode days. The temporal and spatial distribution of the PBLH in Maryland was modeled with the Weather Research and Forecasting (WRF) model to characterize the role of shoreline circulation and thermally-induced boundary layers on locations along the Chesapeake Bay. National Research Council (2009), Observing Weather and Climate from the Ground Up: A

  6. Adsorption of Human Tear Lipocalin to Human Meibomian Lipid Films

    PubMed Central

    Millar, Thomas J.; Mudgil, Poonam; Butovich, Igor A.; Palaniappan, Chendur K.

    2009-01-01

    Purpose Tear lipocalin (Tlc) is a major lipid binding protein in tears and is thought to have an important role in stabilizing the Meibomian lipid layer by transferring lipids to it from the aqueous layer or ocular surface, or by adsorbing to it directly. These possible roles have been investigated in vitro using human Tlc. Methods Tlc was purified from human tears by size exclusion chromatography followed by ion exchange chromatography. Three additional samples of the Tlc were prepared by lipidation, delipidation, and relipidation. The lipids extracted from the purified Tlc were analyzed by HPLC-MS followed by fragmentation. Adsorption of these different forms of Tlc to a human Meibomian lipid film spread on the surface of an artificial tear buffer in a Langmuir trough were observed by recording changes in the pressure with time (∏-T profile) and monitoring the appearance of the film microscopically. These results were compared with similar experiments using a bovine Meibomian lipid film. Results The results indicated that Tlc binds slowly to a human Meibomian lipid film compared with lysozyme or lactoferrin, even at 37°C. The adsorption of Tlc to a human Meibomian lipid film was very different from its adsorption to a bovine Meibomian lipid film, indicating the nature of the lipids in the film is critical to the adsorption process. Similarly, the different forms of Tlc had quite distinct adsorption patterns, as indicated both by changes in ∏-T profiles and the microscopic appearance of the films. Conclusions It was concluded that human Tlc was capable of adsorbing to and penetrating into a Meibomian lipid layer, but this process is very complex and depends on both the types of lipids bound to Tlc and the lipid complement comprising the Meibomian lipid film. PMID:18757516

  7. Hydrophobic Surface Burial is the Major Stability Determinant of a Flat, Single-Layer β-Sheet

    PubMed Central

    Yan, Shude; Gawlak, Grzegorz; Makabe, Koki; Tereshko, Valentina; Koide, Akiko; Koide, Shohei

    2007-01-01

    Formation of a flat β-sheet is a fundamental event in β-sheet-mediated protein self-assembly. To investigate contributions of various factors to the stability of flat β-sheets, we performed extensive alanine-scanning mutagenesis experiments on the single-layer β-sheet segment of Borrelia outer surface protein A (OspA). This β-sheet segment consists of β-strands with highly regular geometries that can serve as a building block for self-assembly. Our Ala-scanning approach is distinct from the conventional host-guest method in that it introduces only conservative, truncation mutations that should minimize structural perturbation. Our results showed very weak correlation with experimental β-sheet propensity scales, statistical β-sheet propensity scales, or cross-strand pairwise correlations. In contrast, our data showed strong positive correlation with the change in buried nonpolar surface area. Polar interactions including prominent Glu-Lys cross-strand pairs marginally contribute to the β-sheet stability. These results were corroborated by results from additional non-Ala mutations. Taken together, these results demonstrate the dominant contribution of nonpolar surface burial to flat β-sheet stability even at solvent-exposed positions. The OspA single-layer β-sheet achieves efficient hydrophobic surface burial without forming a hydrophobic core by a strategic placement of a variety of side chains. These findings further suggest the importance of hydrophobic interactions within a β-sheet layer in peptide self-assembly. PMID:17335845

  8. Structure determination of functional membrane proteins using small-angle neutron scattering (sans) with small, mixed-lipid liposomes: native beef heart mitochondrial cytochrome c oxidase forms dimers.

    PubMed

    Rubinson, Kenneth A; Pokalsky, Christine; Krueger, Susan; Prochaska, Lawrence J

    2013-01-01

    The low-resolution three-dimensional structure of purified native beef heart mitochondrial cytochrome c oxidase (COX) in asolectin unilamellar liposomes has been measured by small-angle neutron scattering under the conditions where the protein remains fully functional. From a neutron scattering perspective, the use of mixed-lipid liposomes provided for a more homogeneous matrix than can be achieved using a single lipid. As a result, the measurements were able to be performed under conditions where the liposome scattering was essentially eliminated (contrast-matched conditions). The protein structure in the membrane was modeled as a simple parallelepiped with side lengths of (59 × 70 × 120) Å with uncertainties, respectively, (11, 12, 20 Å). The molecular mass calculated for a typical protein with this volume is estimated to be (410 ± 124) kDa, which indicates the mass of a COX dimer. The longest dimension has some uncertainty due to intermolecular scattering contributing to the data. Nevertheless, that length was estimated using an average protein density and the known dimer molecular mass. Using the same cross sectional dimensions for the structure, the length is estimated to be 120 Å. However, the measured scattering curve of the dimer in the liposome differs significantly from that calculated from the X-ray structure of the dimer in a crystal of mixed micelles (PDB 3AG1). The calculated SANS scattering from the crystal structure was fit with a parallelepiped, measuring (59 × 101 × 129) Å with fitting uncertainties, respectively, (2, 3, 3 Å). Our results suggest that COX is a functional dimer when reconstituted into mixed-lipid liposomes.

  9. Tocopheryl acetate nanoemulsions stabilized with lipid-polymer hybrid emulsifiers for effective skin delivery.

    PubMed

    Nam, Yoon Sung; Kim, Jin-Woong; Park, Jaeyoon; Shim, Jongwon; Lee, Jong Suk; Han, Sang Hoon

    2012-06-01

    Tocopheryl acetate is used as the oil component of nanoemulsions using a mixture of unsaturated phospholipids and polyethylene oxide-block-poly(ε-caprolactone) (PEO-b-PCL). This study investigates the effects of the lipid-polymer composition on the size and surface charge of nanoemulsions, microviscosity of the interfacial layer, and skin absorption of tocopheryl acetate. The lipid-polymer hybrid system exhibits excellent colloidal dispersion stability, which is comparable to that of polymer-based nanoemulsions. If lipids are used as emulsifiers, nanoemulsions show poor dispersion stability despite a good skin absorption enhancing effect. The amount of tocopheryl acetate absorbed by the skin increases with an increased lipid-to-polymer ratio, as determined using the hairless guinea pig skin loaded in a Franz-type diffusion cell. An 8:2 (w/w) mixture of unsaturated phospholipids and PEO-b-PCL exhibits the most efficient delivery of tocopheryl acetate into the skin. Our results show that tocopheryl acetate is absorbed almost twice as fast by the lipid-polymer hybrid system than the nanoemulsions stabilized with PEO-b-PCL. This study suggests that the lipid-polymer hybrid system can be used as an effective means of optimizing nanoemulsions in terms of dispersion stability and skin delivery capability.

  10. Microfluidic fabrication of asymmetric giant lipid vesicles

    PubMed Central

    Hu, Peichi C.; Li, Su; Malmstadt, Noah

    2011-01-01

    We have developed a microfluidic technology for the fabrication of compositionally asymmetric giant unilamellar vesicles (GUVs). The vesicles are assembled in two independent steps. In each step, a lipid monolayer is formed at a water-oil interface. The first monolayer is formed inside of a microfluidic device with a multiphase droplet flow configuration consisting of a continuous oil stream in which water droplets are formed. These droplets are dispensed into a vessel containing a layer of oil over a layer of water. The second lipid monolayer is formed by transferring the droplets through this second oil-water interface by centrifugation. By dissolving different lipid compositions in the different oil phases, the composition of each leaflet of the resulting lipid bilayer can be controlled. We have demonstrated membrane asymmetry by showing differential fluorescence quenching of labeled lipids in each leaflet and by demonstrating that asymmetric GUVs will bind an avidin-coated surface only when biotinylated lipids are targeted to the outer leaflet. In addition, we have demonstrated the successful asymmetric targeting of phosphatidylserine lipids to each leaflet, producing membranes with a biomimetic and physiologically relevant compositional asymmetry. PMID:21449588

  11. Label-free determination of lipid composition and secondary protein structure of human salivary noncancerous and cancerous tissues by Raman microspectroscopy.

    PubMed

    Brozek-Pluska, Beata; Kopec, Monika; Niedzwiecka, Izabela; Morawiec-Sztandera, Alina

    2015-04-01

    The applications of optical spectroscopic methods in cancer detection open new possibilities in oncological diagnostics. Raman spectroscopy and Raman imaging represent noninvasive, label-free, and rapidly developing tools in cancer diagnosis. In the study described in this paper Raman microspectroscopy has been employed to examine noncancerous and cancerous human salivary gland tissues of the same patient. The most significant differences between noncancerous and cancerous tissues were found in regions typical for the vibrations of lipids and proteins. The detailed analysis of secondary structures of proteins contained in the cancerous and the noncancerous tissues is also presented.

  12. Determination of the shear deformation-potential constant in electron space-charge layers on Si surfaces by tunneling

    NASA Astrophysics Data System (ADS)

    Kunze, U.

    1985-10-01

    The influence of uniaxial mechanical stress on the subband structure in electron inversion layers has been studied in metal-SiO2-Si(001) junctions by means of tunneling spectroscopy. Compressive or tensile stresses applied in the [110] direction result respectively in a uniform lowering or raising of the energy of the higher primed subband system against the lower one. For stresses applied along [100] a splitting of each of the fourfold-degenerate primed levels into two levels is observed. Taking into account the bias dependence of the subband energies and the voltage drop across the channel series resistance, the tunneling characteristics yield a value for the shear deformation-potential constant Ξu=8.6+/-0.4 eV that is in accordance with the known bulk value. An extrapolation of the energy splitting between subbands belonging to the [100] and [010] valley pairs to zero stress along [100] reveals a preexisting splitting of 0.5 meV, which corresponds to a residual stress in the surface layer of 6 N mm-2.

  13. Lipid14: The Amber Lipid Force Field.

    PubMed

    Dickson, Callum J; Madej, Benjamin D; Skjevik, Age A; Betz, Robin M; Teigen, Knut; Gould, Ian R; Walker, Ross C

    2014-02-11

    The AMBER lipid force field has been updated to create Lipid14, allowing tensionless simulation of a number of lipid types with the AMBER MD package. The modular nature of this force field allows numerous combinations of head and tail groups to create different lipid types, enabling the easy insertion of new lipid species. The Lennard-Jones and torsion parameters of both the head and tail groups have been revised and updated partial charges calculated. The force field has been validated by simulating bilayers of six different lipid types for a total of 0.5 μs each without applying a surface tension; with favorable comparison to experiment for properties such as area per lipid, volume per lipid, bilayer thickness, NMR order parameters, scattering data, and lipid lateral diffusion. As the derivation of this force field is consistent with the AMBER development philosophy, Lipid14 is compatible with the AMBER protein, nucleic acid, carbohydrate, and small molecule force fields. PMID:24803855

  14. Electrostatic modulation and enzymatic cross-linking of interfacial layers impacts gastrointestinal fate of multilayer emulsions.

    PubMed

    Zeeb, Benjamin; Weiss, Jochen; McClements, David Julian

    2015-08-01

    In this study, membrane properties were modulated using layer-by-layer electrostatic depositioning in combination with salt and/or enzyme treatment to control the gastrointestinal fate of emulsified oils. Lipid droplets coated by a single-layer of biopolymers (gelatin) were prepared by high pressure homogenization. Lipid droplets coated by a double-layer of biopolymers (gelatin-pectin) were prepared by electrostatically depositing sugar beet pectin on the gelatin-coated droplets. Laccase was added to the double-layer emulsions to covalently crosslink the adsorbed pectin molecules, whereas sodium chloride was added to modulate interfacial properties through electrostatic screening effects. Non-cross-linked and cross-linked double-layer emulsions (with and without salt) were then passed through a simulated gastrointestinal tract (GIT) that included mouth, gastric and intestinal phases. Free fatty acid release profiles suggested that the stability of the emulsified droplets within the GIT played a more important role in determining the rate and extent of lipid digestion than the initial interfacial layer properties.

  15. Electrostatic modulation and enzymatic cross-linking of interfacial layers impacts gastrointestinal fate of multilayer emulsions.

    PubMed

    Zeeb, Benjamin; Weiss, Jochen; McClements, David Julian

    2015-08-01

    In this study, membrane properties were modulated using layer-by-layer electrostatic depositioning in combination with salt and/or enzyme treatment to control the gastrointestinal fate of emulsified oils. Lipid droplets coated by a single-layer of biopolymers (gelatin) were prepared by high pressure homogenization. Lipid droplets coated by a double-layer of biopolymers (gelatin-pectin) were prepared by electrostatically depositing sugar beet pectin on the gelatin-coated droplets. Laccase was added to the double-layer emulsions to covalently crosslink the adsorbed pectin molecules, whereas sodium chloride was added to modulate interfacial properties through electrostatic screening effects. Non-cross-linked and cross-linked double-layer emulsions (with and without salt) were then passed through a simulated gastrointestinal tract (GIT) that included mouth, gastric and intestinal phases. Free fatty acid release profiles suggested that the stability of the emulsified droplets within the GIT played a more important role in determining the rate and extent of lipid digestion than the initial interfacial layer properties. PMID:25766826

  16. Validation Thin Layer Chromatography for the Determination of Acetaminophen in Tablets and Comparison with a Pharmacopeial Method

    PubMed Central

    Pyka, Alina; Budzisz, Marika; Dołowy, Małgorzata

    2013-01-01

    Adsorption thin layer chromatography (NP-TLC) with densitometry has been established for the identification and the quantification of acetaminophen in three leading commercial products of pharmaceutical tablets coded as brand: P1 (Product no. 1), P2 (Product no. 2), and P3 (Product no. 3). Applied chromatographic conditions have separated acetaminophen from its related substances, namely, 4-aminophenol and and 4′-chloroacetanilide. UV densitometry was performed in absorbance mode at 248 nm. The presented method was validated by specificity, range, linearity, accuracy, precision, detection limit, quantitative limit, and robustness. The TLC-densitometric method was also compared with a pharmacopeial UV-spectrophotometric method for the assay of acetaminophen, and the results confirmed statistically that the NP-TLC-densitometric method can be used as a substitute method. It could be said that the validated NP-TLC-densitometric method is suitable for the routine analysis of acetaminophen in quantity control laboratories. PMID:24063006

  17. C and N depth profiles of SiCN layers determined with nuclear reaction analyses and AES

    NASA Astrophysics Data System (ADS)

    Link, F.; Baumann, H.; Bethge, K.; Klewe-Nebenius, H.; Bruns, M.

    1998-04-01

    Si 1C xN y layers were prepared by sequential implantation of 40 keV 13C- and 50 keV 15N-ions into c-Si <1 1 1> samples near RT. The carbon and nitrogen depth distributions were measured using the resonant nuclear (p,γ) reactions 15N(p,αγ) 12C at Eres=429 keV and 13C(p,γ) 14N at Eres=1748 keV, respectively. The measured raw data of depth profiling (gamma yield versus the proton beam energy) are converted to concentration-depth profiles of the elements C, N and Si with a common depth scale by using a new developed computer algorithm. These concentration profiles are compared with those obtained with Auger Electron Spectroscopy (AES) and non-Rutherford Backscattering Spectrometry (n-RBS).

  18. Using of standard marine radar for determination of a water surface and an atmosphere near-surface layer parameters

    NASA Astrophysics Data System (ADS)

    Bogatov, Nikolay A.; Bakhanov, Victor V.; Ermoshkin, Aleksei V.; Kazakov, Vasily I.; Kemarskaya, Olga N.; Titov, Victor I.; Troitskaya, Yulia I.

    2014-10-01

    At present time radar methods of the seas and oceans diagnostics are actively developing. Using of the radar stations based on satellites and planes allows to receive information on a sea surface and a atmosphere near-surface layer with coverage of big water surface areas independently of day time. The developed methods of satellite radio images processing can be applied to marine radar stations. In Institute of Applied Physics RAS works on sea surface diagnostics systems development on the basis of standard marine radar are actively conducted. Despite smaller coverage of the territory in comparison with satellite data, marine radar have possibility to record spatially temporary radar images and to receive information on a surrounding situation quickly. This work deals with results of the researches which were conducted within the international expedition in the Atlantic Ocean in the autumn of 2012 on a route Rotterdam (Netherlands) - Ushuaya (Argentina) - Antarctica — Ushuaya. During this expedition a complex measurements of a sea surface, a atmosphere near-surface layer parameters and subsurface currents in the wide range of hydroweather conditions, including the storm were carried out. The system developed in IAP RAS on the basis of a marine radar ICOM MR-1200RII and the ADC (Analog Digital Converter) block for data recording on the personal computer was used. Display of a non-uniform near-surface current on sea surface radar images in storm conditions is shown. By means of the high-speed anemometer and meteorological station the measurements of the atmosphere parameters were carried out. Comparison of the anemometer data with calculated from radar images is carried out. Dependence of radar cross section from wind speed in the wide range of wind speeds, including storm conditions is investigated. Possibility of marine radar using for surface waves intensity and ice situation estimates also as icebergs detection is shown.

  19. X-ray attenuation measurements in a cavitating mixing layer for instantaneous two-dimensional void ratio determination

    SciTech Connect

    Aeschlimann, Vincent; Barre, Stephane; Legoupil, Samuel

    2011-05-15

    The purpose of this experimental study was to analyze a two-dimensional cavitating shear layer. The global aim of this work was to obtain a better understanding and modeling of cavitation phenomenon in a 2D turbulent sheared flow which can be considered as quite representative of cavitating rocket engine turbopomp inducers. This 2D mixing layer flow provided us a well documented test case which can be used for the characterization of the cavitation effects in sheared flows. The development of a velocity gradient was observed inside a liquid water flow: Kelvin-Helmholtz instabilities developed at the interface. Vaporizations and implosions of cavitating structures inside the vortices were observed. X-ray attenuation measurements were performed to estimate the amount of vapor present inside the mixing area. Instantaneous two-dimensional void ratio fields were acquired. The real spatial resolutions are 0.5 mm with 2000 fps and 1.5 mm with 20 000 fps. The effective time resolution is equal to the camera frame rate up to a 19% void ratio variation between two consecutive images. This seems to be sufficient in the context of the present flow configuration. The two-phase structures present inside the mixing area were analyzed at three different cavitation levels and their behaviors were compared to non-cavitating flow dynamic. Convection velocities and vortices shedding frequencies were estimated. Results show that vapor was transported by the turbulent velocity field. Statistical analysis of the void ratio signal was carried out up to the fourth order moment. This study provided a global understanding of the cavitating structure evolution and of the cavitation effects on turbulent sheared flows.

  20. Paleointensity determination of welded tuffs extruded with tephra layers: A new approach to calibration of relative paleointensity stacks

    NASA Astrophysics Data System (ADS)

    Mochizuki, N.; Fujii, S.; Hasegawa, T.; Yamamoto, Y.; Hatakeyama, T.; Okada, M.; Shibuya, H.

    2015-12-01

    For a reliable calibration of a relative paleointensity stack, we proposed a new method for direct comparison of absolute paleointensities (APIs) with relative paleointensities (RPIs) (Mochizuki et al., under review). In the analysis, APIs are directly compared with the RPIs of a RPI stack at six stratigraphic levels: three levels are based on tephrostratigraphic correlations between welded tuffs and corresponding tephra layers in the oxygen isotope stratigraphy, and the other three levels are based on paleomagnetic correlations between RPI minima and transitional geomagnetic fields. In the present study, to increase API data with tephrostratigraphic correlation, we applied the LTD-DHT Shaw paleointensity method (Tsunakawa-Shaw method) to 21 welded tuffs in Japan extruded with widespread tephra layers. We obtained mean paleointensities for 16 of the 21 welded tuffs. Since nine of the 16 welded tuffs were correlated with the tephras recognized in the oxygen isotope stratigraphy, they can be added to the API data used in the direct comparison method. Combining these API data with the reported data, we compared API data with RPIs from the PISO-1500 stack and SINT-800 stack at the 14 stratigraphic levels: eleven levels are based on tephrostratigraphic correlation and the other three levels are based on paleomagnetic correlation. RPIs of the PISO-1500 stack showed a linear relationship with the virtual axial dipole moments (VADMs) calculated from the APIs, indicating that the PISO-1500 stack has a linear relation to the axial dipole moment. PRIs from the SINT-800 stack also have a linear-like trend with the VADMs. This direct comparison method can clarify the relationship between APIs and RPIs of a RPI stack, and thus provide a reliable calibration of the RPI stack to absolute values.

  1. A Eukaryotic Sensor for Membrane Lipid Saturation.

    PubMed

    Covino, Roberto; Ballweg, Stephanie; Stordeur, Claudius; Michaelis, Jonas B; Puth, Kristina; Wernig, Florian; Bahrami, Amir; Ernst, Andreas M; Hummer, Gerhard; Ernst, Robert

    2016-07-01

    Maintaining a fluid bilayer is essential for cell signaling and survival. Lipid saturation is a key factor determining lipid packing and membrane fluidity, and it must be tightly controlled to guarantee organelle function and identity. A dedicated eukaryotic mechanism of lipid saturation sensing, however, remains elusive. Here we show that Mga2, a transcription factor conserved among fungi, acts as a lipid-packing sensor in the ER membrane to control the production of unsaturated fatty acids. Systematic mutagenesis, molecular dynamics simulations, and electron paramagnetic resonance spectroscopy identify a pivotal role of the oligomeric transmembrane helix (TMH) of Mga2 for intra-membrane sensing, and they show that the lipid environment controls the proteolytic activation of Mga2 by stabilizing alternative rotational orientations of the TMH region. This work establishes a eukaryotic strategy of lipid saturation sensing that differs significantly from the analogous bacterial mechanism relying on hydrophobic thickness. PMID:27320200

  2. The Effect of Cholesterol on Short- and Long-Chain Monounsaturated Lipid Bilayers as Determined by Molecular Dynamics Simulations and X-Ray Scattering

    PubMed Central

    Kučerka, Norbert; Perlmutter, Jason D.; Pan, Jianjun; Tristram-Nagle, Stephanie; Katsaras, John; Sachs, Jonathan N.

    2008-01-01

    We investigate the structure of cholesterol-containing membranes composed of either short-chain (diC14:1PC) or long-chain (diC22:1PC) monounsaturated phospholipids. Bilayer structural information is derived from all-atom molecular dynamics simulations, which are validated via direct comparison to x-ray scattering experiments. We show that the addition of 40 mol % cholesterol results in a nearly identical increase in the thickness of the two different bilayers. In both cases, the chain ordering dominates over the hydrophobic matching between the length of the cholesterol molecule and the hydrocarbon thickness of the bilayer, which one would expect to cause a thinning of the diC22:1PC bilayer. For both bilayers there is substantial headgroup rearrangement for lipids directly in contact with cholesterol, supporting the so-called umbrella model. Importantly, in diC14:1PC bilayers, a dynamic network of hydrogen bonds stabilizes long-lived reorientations of some cholesterol molecules, during which they are found to lie perpendicular to the bilayer normal, deep within the bilayer's hydrophobic core. Additionally, the simulations show that the diC14:1PC bilayer is significantly more permeable to water. These differences may be correlated with faster cholesterol flip-flop between the leaflets of short-chain lipid bilayers, resulting in an asymmetric distribution of cholesterol molecules. This asymmetry was observed experimentally in a case of unilamellar vesicles (ULVs), and reproduced through a set of novel asymmetric simulations. In contrast to ULVs, experimental data for oriented multilamellar stacks does not show the asymmetry, suggesting that it results from the curvature of the ULV bilayers. PMID:18515383

  3. [Determination of the four generic fractions of aged bitumen by thin-layer chromatography with flame ionization detection].

    PubMed

    Feng, Zhengang; Zhang, Jianbin; Li, Xinjun; Yu, Jianying

    2015-02-01

    The aging process of bitumen has been paid more and more attention by the researchers. The four generic fractions (saturates, aromatics, resins and asphaltenes) of bitumen change significantly during the aging process. The analysis of the changes of the four generic fractions of bitumen is very helpful to reveal the bitumen aging mechanisms and guide its engineering applications. In this study, the bitumen was aged by thin film oven test (TFOT) , pressurized aging vessel (PAV) test and ultraviolet (UV) aging test, respectively. Then the four generic fractions of bitumen before and after aging were analyzed by thin-layer chromatography with flame ionization detection (TLC-FID) , which was further compared with the solubility procedures and chromatographic technique ( named as Corbett method). The compositions of the expanded solvents were also investigated. Finally, the correlation between the TLC-FID and Corbett method was further studied, which revealed a proper TLC-FID meth- od for detection of aged bitumen. The bitumen solution dissolved by dichloromethane was successively expanded by n-heptane, toluene/n-heptane (80 :20, v/v) and toluene/ethanol (55: 45, v/v) , followed by TLC-FID. This method is of great significance for the analysis of the four generic fractions of bitumen and for the exploration of bitumen aging mechanisms. PMID:25989694

  4. Nucleation and initial growth of atomic layer deposited titanium oxide determined by spectroscopic ellipsometry and the effect of pretreatment by surface barrier discharge

    NASA Astrophysics Data System (ADS)

    Cameron, David C.; Krumpolec, Richard; Ivanova, Tatiana V.; Homola, Tomáš; Černák, Mirko

    2015-08-01

    This paper reports on the use of spectroscopic ellipsometry to characterise the initial nucleation stage of the atomic layer deposition of the anatase phase of titanium dioxide on silicon substrates. Careful control and analysis of the ellipsometric measurements enables the determination of the evolution of crystallite diameter and surface density in the nucleation stage before a continuous film is formed. This growth behaviour is in line with atomic force microscopy measurements of the crystallite size. The crystallite diameter is a linear function of the number of ALD cycles with a slope of approximately 1.7 Å cycle-1 which is equivalent to a layer growth rate of 0.85 Å cycle-1 consistent with a ripening process which increases the crystallite size while reducing their density. The crystallite density decreases from ∼3 × 1017 m-3 in the initial nucleation stages to ∼3 × 1015 m-3 before the film becomes continuous. The effect of exposing the substrate to a diffuse coplanar surface barrier discharge in an air atmosphere before deposition was measured and only small differences were found: the plasma treated samples were slightly rougher in the initial stages and required a greater number of cycles to form a continuous film (∼80) compared to the untreated films (∼50). A thicker layer of native oxide was found after plasma treatment.

  5. Comparison of thickness, grade, and depth of radioactive layers as determined by gamma-ray logging and by core sampling

    USGS Publications Warehouse

    Bunker, Carl M.

    1959-01-01

    Thickness, grade, and depth data were obtained by analyzing gamma-ray logs and core samples from 56 diamond drill holes penetrating uranium deposits in the Colorado Plateau. The data from the two methods were compared to determine variations found in gamma-ray log interpretation and chemical and radiometric analyses of the drill core. Correlations within each parameter varied among the drilling areas analyzed. Gamma-ray interpretations of grade compared to chemical analyses were within the range of -10 to +25 percent. Most depth measurements determined by gamma-ray log interpretation compared to drill core measurement were within 0.5 percent. Results of the study indicate a need for better thickness definition in both gamma-ray logging and core scanning equipment.

  6. A rapid method for determining arachidonic:eicosapentaenoic acid ratios in whole blood lipids: correlation with erythrocyte membrane ratios and validation in a large Italian population of various ages and pathologies

    PubMed Central

    2010-01-01

    Background Omega-3 and -6 polyunsaturated fatty acids (LCPUFA), are important for good health conditions. They are present in membrane phospholipids. The ratio of total n-6:n-3 LCPUFA and arachidonic acid:eicosapentaenoic acid (AA and EPA), should not exceed 5:1. Increased intake of n-6 and decreased consumption of n-3 has resulted in much higher, ca 10/15:1 ratio in RBC fatty acids with the possible appearance of a pathological "scenario". The determination of RBC phospholipid LCPUFA contents and ratios is the method of choice for assessing fatty acid status but it is labour intensive and time consuming. Aims of the study [i] To describe and validate a rapid method, suitable for large scale population studies, for total blood fatty acid assay; [ii] to verify a possible correlation between total n-6:n-3 ratio and AA:EPA ratios in RBC phospholipids and in whole-blood total lipids, [iii] to assess usefulness of these ratio as biomarkers of LCPUFA status. Methods [1] Healthy volunteers and patients with various pathologies were recruited. [2] Fatty acid analyses by GC of methyl esters from directly derivatized whole blood total lipids and from RBC phospholipids were performed on fasting blood samples from 1432 subjects categorised according to their age, sex and any existing pathologies. AA:EPA ratio and the total n-6:n-3 ratio were determined. Results AA:EPA ratio is a more sensitive and reliable index for determining changes in total blood fatty acid and it is correlated with the ratio derived from extracted RBC phospholipids. Conclusions The described AA:EPA ratio is a simple, rapid and reliable method for determining n-3 fatty acid status. PMID:20105293

  7. Purification of vetiver alcohols and esters for quantitative high-performance thin-layer chromatography determination in Haitian vetiver essential oils and vetiver acetates.

    PubMed

    Paillat, Lionel; Périchet, Christine; Pierrat, Jean-Philippe; Lavoine, Sophie; Filippi, Jean-Jacques; Meierhenrich, Uwe; Fernandez, Xavier

    2012-06-01

    A simple, fast, and efficient High-Performance Thin-Layer Chromatography (HPTLC) method was developed for the simultaneous quantitative determination of alcohols and acetates in Haitian vetiver essential oils (Chryzopogon zizanioides) and its acetylated form. Analytes were separated by using a mixture of n-hexane-chloroform-ethyl acetate (8:6:0.5, v/v/v) as mobile phase under 47% humidity. Quantification was achieved by densitometric evaluation of the analytes in absorbance mode under visible light (λ=530 nm) after staining with a vanillin-sulfuric acid reagent. Reference mixtures of alcohols and acetates were obtained by fractionation of Haitian vetiver oil or vetiver acetates, followed by purification of the fractions of interest by means of Over-Pressured Layer Chromatography (OPLC). The chemical composition of each reference fraction was determined by using GC-MS and GC×GC-MS, and their overall purity was determined by GC/FID and HPTLC. The TLC method provided compact spots for alcohols (R(f2)=0.18±0.01 and R(f1)=0.28±0.01) and acetates (R(f3)=0.65±0.01). Calibration plots showed good linear correlation with r²=0.9995±0.0001 and r²=0.9995±0.0001 for alcohols and r²=0.9996±0.0001 for acetates in a 40-200 ng spot⁻¹ concentration range with respect to peak areas. The method was validated for precision and accuracy. Limit of detection (LOD) and quantification (LOQ) were determined. Method specificity was confirmed using retention factor (R(f)) and GC-MS control of the standards reference mixtures. PMID:22560705

  8. Determination of layer thickness and optical constants of thin films by using a modified pattern search method.

    PubMed

    Miloua, R; Kebbab, Z; Chiker, F; Sahraoui, K; Khadraoui, M; Benramdane, N

    2012-02-15

    We propose the use of a pattern search optimization technique in combination with a seed preprocessing procedure to determine the optical constants and thickness of thin films using only the transmittance spectra. The approach is quite flexible, straightforward to implement, and efficient in reaching the best fitting. We demonstrate the effectiveness of the method in extracting optical constants, even when the films are not displaying interference fringes. Comparison to a real-coded genetic algorithm shows that the modified pattern search is fast, almost accurate, and does not need any parameter adjustments. The approach is successfully applied to extract the thickness and optical constants of spray pyrolyzed nanocrystalline CdO thin films.

  9. Raman determination of layer stresses and strains for heterostructures and its application to the cubic SiC/Si system

    NASA Technical Reports Server (NTRS)

    Feng, Z. C.; Choyke, W. J.; Powell, J. A.

    1988-01-01

    A set of formulas for a generalized axial stress in diamond and zinc-blende semiconductors under axial stress is derived to calculate stress-related Raman shifts. By analyzing known Raman data on cubic SiC under hydrostatic pressures, one of the Raman-stress coefficients was obtained, and the existing elastic stiffness constants of cubic SiC were optimized. A method for calculating the stress and strain in SiC films on (100) Si is proposed. It is suggested that the stress and strain expressions and the method of the stress and strain determinations in heterostructures are quite general and may be used for other systems.

  10. Disorders of Lipid Metabolism

    MedlinePlus

    ... Metabolic Disorders Disorders of Carbohydrate Metabolism Disorders of Amino Acid Metabolism Disorders of Lipid Metabolism Fats (lipids) are ... carbohydrates and low in fats. Supplements of the amino acid carnitine may be helpful. The long-term outcome ...

  11. Irinotecan Lipid Complex Injection

    MedlinePlus

    Irinotecan lipid complex is used in combination with other medications to treat pancreatic cancer that has spread to other parts of ... after treatment with other chemotherapy medications. Irinotecan lipid complex is in a class of antineoplastic medications called ...

  12. Vincristine Lipid Complex Injection

    MedlinePlus

    Vincristine lipid complex is used to treat a certain type of acute lymphoblastic leukemia (ALL; a type of cancer of the ... two different treatments with other medications. Vincristine lipid complex is in a class of medications called vinca ...

  13. Daunorubicin Lipid Complex Injection

    MedlinePlus

    Daunorubicin lipid complex is used to treat advanced Kaposi's sarcoma (a type of cancer that causes abnormal tissue to grow on ... related to acquired immunodeficiency syndrome (AIDS). Daunorubicin lipid complex is in a class of medications called anthracyclines. ...

  14. Cytarabine Lipid Complex Injection

    MedlinePlus

    Cytarabine lipid complex is used to treat lymphomatous meningitis (a type of cancer in the covering of the spinal cord and brain). Cytarabine lipid complex is in a class of medications called antimetabolites. ...

  15. Doxorubicin Lipid Complex Injection

    MedlinePlus

    Doxorubicin lipid complex is used to treat ovarian cancer that has not improved or that has worsened after treatment with other medications. Doxorubicin lipid complex is also used to treat Kaposi's sarcoma (a ...

  16. Skin lipids from Saudi Arabian birds.

    PubMed

    Khan, Haseeb A; Arif, Ibrahim A; Williams, Joseph B; Champagne, Alex M; Shobrak, Mohammad

    2014-04-01

    Skin lipids play an important role in the regulation of cutaneous water loss (CWL). Earlier studies have shown that Saudi desert birds exhibit a tendency of reduced CWL than birds from temperate environment due to adaptive changes in composition of their skin lipids. In this study, we used thin-layer chromatography (TLC) for separation and detection of non-polar and polar lipids from the skin of six bird species including sooty gull, brown booby, house sparrow, Arabian waxbill, sand partridge, and laughing dove. The lipids were separated and detected on Silica gel G coated TLC plates and quantified by using densitometric image analysis. Rf values of the non-polar lipids were as follows: cholesterol (0.29), free fatty acids (0.58), triacylglycerol (0.69), fatty acids methyl esters (0.84) and cholesterol ester (0.97). Rf values for the polar lipids were: cerebroside (0.42), ceramide (0.55) and cholesterol (0.73). The results showed the abundance of fatty acids methyl esters (47.75-60.46%) followed by triacylglycerol (12.69-24.14%). The remaining lipid compositions were as follows: cholesterol (4.09-13.18%), ceramide (2.18-13.27%), and cerebroside (2.53-12.81%). In conclusion, our findings showed that TLC is a simple and sensitive method for the separation and quantification of skin lipids. We also reported a new protocol for lipid extraction using the zirconia beads for efficient disruption of skin tissues. This study will help us better understand the role of skin lipids in adaptive physiology towards adverse climatic conditions.

  17. Skin lipids from Saudi Arabian birds.

    PubMed

    Khan, Haseeb A; Arif, Ibrahim A; Williams, Joseph B; Champagne, Alex M; Shobrak, Mohammad

    2014-04-01

    Skin lipids play an important role in the regulation of cutaneous water loss (CWL). Earlier studies have shown that Saudi desert birds exhibit a tendency of reduced CWL than birds from temperate environment due to adaptive changes in composition of their skin lipids. In this study, we used thin-layer chromatography (TLC) for separation and detection of non-polar and polar lipids from the skin of six bird species including sooty gull, brown booby, house sparrow, Arabian waxbill, sand partridge, and laughing dove. The lipids were separated and detected on Silica gel G coated TLC plates and quantified by using densitometric image analysis. Rf values of the non-polar lipids were as follows: cholesterol (0.29), free fatty acids (0.58), triacylglycerol (0.69), fatty acids methyl esters (0.84) and cholesterol ester (0.97). Rf values for the polar lipids were: cerebroside (0.42), ceramide (0.55) and cholesterol (0.73). The results showed the abundance of fatty acids methyl esters (47.75-60.46%) followed by triacylglycerol (12.69-24.14%). The remaining lipid compositions were as follows: cholesterol (4.09-13.18%), ceramide (2.18-13.27%), and cerebroside (2.53-12.81%). In conclusion, our findings showed that TLC is a simple and sensitive method for the separation and quantification of skin lipids. We also reported a new protocol for lipid extraction using the zirconia beads for efficient disruption of skin tissues. This study will help us better understand the role of skin lipids in adaptive physiology towards adverse climatic conditions. PMID:24600311

  18. Skin lipids from Saudi Arabian birds

    PubMed Central

    Khan, Haseeb A.; Arif, Ibrahim A.; Williams, Joseph B.; Champagne, Alex M.; Shobrak, Mohammad

    2013-01-01

    Skin lipids play an important role in the regulation of cutaneous water loss (CWL). Earlier studies have shown that Saudi desert birds exhibit a tendency of reduced CWL than birds from temperate environment due to adaptive changes in composition of their skin lipids. In this study, we used thin-layer chromatography (TLC) for separation and detection of non-polar and polar lipids from the skin of six bird species including sooty gull, brown booby, house sparrow, Arabian waxbill, sand partridge, and laughing dove. The lipids were separated and detected on Silica gel G coated TLC plates and quantified by using densitometric image analysis. Rf values of the non-polar lipids were as follows: cholesterol (0.29), free fatty acids (0.58), triacylglycerol (0.69), fatty acids methyl esters (0.84) and cholesterol ester (0.97). Rf values for the polar lipids were: cerebroside (0.42), ceramide (0.55) and cholesterol (0.73). The results showed the abundance of fatty acids methyl esters (47.75–60.46%) followed by triacylglycerol (12.69–24.14%). The remaining lipid compositions were as follows: cholesterol (4.09–13.18%), ceramide (2.18–13.27%), and cerebroside (2.53–12.81%). In conclusion, our findings showed that TLC is a simple and sensitive method for the separation and quantification of skin lipids. We also reported a new protocol for lipid extraction using the zirconia beads for efficient disruption of skin tissues. This study will help us better understand the role of skin lipids in adaptive physiology towards adverse climatic conditions. PMID:24600311

  19. Nuclear lipid droplets: a novel nuclear domain.

    PubMed

    Layerenza, J P; González, P; García de Bravo, M M; Polo, M P; Sisti, M S; Ves-Losada, A

    2013-02-01

    We investigated nuclear neutral-lipid (NL) composition and organization, as NL may represent an alternative source for providing fatty acids and cholesterol (C) to membranes, signaling paths, and transcription factors in the nucleus. We show here that nuclear NL were organized into nonpolar domains in the form of nuclear-lipid droplets (nLD). By fluorescent confocal microscopy, representative nLD were observed in situ within the nuclei of rat hepatocytes in vivo and HepG2 cells, maintained under standard conditions in culture, and within nuclei isolated from rat liver. nLD were resistant to Triton X-100 and became stained with Sudan Red, OsO4, and BODIPY493/503. nLD and control cytosolic-lipid droplets (cLD) were isolated from rat-liver nuclei and from homogenates, respectively, by sucrose-gradient sedimentation. Lipids were extracted, separated by thin-layer chromatography, and quantified. nLD were composed of 37% lipids and 63% proteins. The nLD lipid composition was as follows: 19% triacylglycerols (TAG), 39% cholesteryl esters, 27% C, and 15% polar lipids; whereas the cLD composition contained different proportions of these same lipid classes, in particular 91% TAG. The TAG fatty acids from both lipid droplets were enriched in oleic, linoleic, and palmitic acids. The TAG from the nLD corresponded to a small pool, whereas the TAG from the cLD constituted the main cellular pool (at about 100% yield from the total homogenate). In conclusion, nLD are a domain within the nucleus where NL are stored and organized and may be involved in nuclear lipid homeostasis. PMID:23098923

  20. Methods for determining the presence of slurry on the crop and in the upper soil layer after application to grassland.

    PubMed

    Rodhe, Lena

    2003-10-01

    Slurry injection in grassland may lead to lower ammonia losses and enhance silage quality compared to surface spreading. In order to evaluate the performance of different slurry injectors, there is a need for methods which measure the degree of crop contamination and the position of the slurry in the soil after spreading. The aim of this study was to identify and test possible methods. The amount of cattle slurry present on the grass stubble after spreading was determined by measuring the electrical conductivity of water used for rinsing grass samples. The slurry placement in the soil profile was measured in cross-sections made through the slurry trails. Methods used were analyses of images taken by different cameras, resistance measurements and by measuring the dimensions of the slurry trails. The method for determining the presence of slurry on the crop was practically applicable and is considered to be reliable. The placement of slurry could not be sufficiently resolved from image taken by the different cameras or by measuring resistance. Instead, visual assessment and measurements of width and depth of the slurry trails gave a good description of the slurry placement in the soil profile.

  1. Bioactive lipid coating of bone allografts directs engraftment and fate determination of bone marrow-derived cells in rat GFP chimeras.

    PubMed

    Das, Anusuya; Segar, Claire E; Chu, Yihsuan; Wang, Tiffany W; Lin, Yong; Yang, Chunxi; Du, Xeujun; Ogle, Roy C; Cui, Quanjun; Botchwey, Edward A

    2015-09-01

    Bone grafting procedures are performed to treat wounds incurred during wartime trauma, accidents, and tumor resections. Endogenous mechanisms of repair are often insufficient to ensure integration between host and donor bone and subsequent restoration of function. We investigated the role that bone marrow-derived cells play in bone regeneration and sought to increase their contributions by functionalizing bone allografts with bioactive lipid coatings. Polymer-coated allografts were used to locally deliver the immunomodulatory small molecule FTY720 in tibial defects created in rat bone marrow chimeras containing genetically-labeled bone marrow for monitoring cell origin and fate. Donor bone marrow contributed significantly to both myeloid and osteogenic cells in remodeling tissue surrounding allografts. FTY720 coatings altered the phenotype of immune cells two weeks post-injury, which was associated with increased vascularization and bone formation surrounding allografts. Consequently, degradable polymer coating strategies that deliver small molecule growth factors such as FTY720 represent a novel therapeutic strategy for harnessing endogenous bone marrow-derived progenitors and enhancing healing in load-bearing bone defects.

  2. Bioactive lipid coating of bone allografts directs engraftment and fate determination of bone marrow-derived cells in rat GFP chimeras

    PubMed Central

    Das, Anusuya; Segar, Claire E.; Chu, Yihsuan; Wang, Tiffany W.; Lin, Yong; Yang, Chunxi; Du, Xeujun; Ogle, Roy C.; Cui, Quanjun; Botchwey, Edward A.

    2015-01-01

    Bone grafting procedures are performed to treat wounds incurred during wartime trauma, accidents, and tumor resections. Endogenous mechanisms of repair are often insufficient to ensure integration between host and donor bone and subsequent restoration of function. We investigated the role that bone marrow-derived cells play in bone regeneration and sought to increase their contributions by functionalizing bone allografts with bioactive lipid coatings. Polymer-coated allografts were used to locally deliver the immunomodulatory small molecule FTY720 in tibial defects created in rat bone marrow chimeras containing genetically-labeled bone marrow for monitoring cell origin and fate. Donor bone marrow contributed significantly to both myeloid and osteogenic cells in remodeling tissue surrounding allografts. FTY720 coatings altered the phenotype of immune cells two weeks post-injury, which was associated with increased vascularization and bone formation surrounding allografts. Consequently, degradable polymer coating strategies that deliver small molecule growth factors such as FTY720 represent a novel therapeutic strategy for harnessing endogenous bone marrow-derived progenitors and enhancing healing in load-bearing bone defects. PMID:26125501

  3. Rewiring Host Lipid Metabolism by Large Viruses Determines the Fate of Emiliania huxleyi, a Bloom-Forming Alga in the Ocean.

    PubMed

    Rosenwasser, Shilo; Mausz, Michaela A; Schatz, Daniella; Sheyn, Uri; Malitsky, Sergey; Aharoni, Asaph; Weinstock, Eyal; Tzfadia, Oren; Ben-Dor, Shifra; Feldmesser, Ester; Pohnert, Georg; Vardi, Assaf

    2014-06-10

    Marine viruses are major ecological and evolutionary drivers of microbial food webs regulating the fate of carbon in the ocean. We combined transcriptomic and metabolomic analyses to explore the cellular pathways mediating the interaction between the bloom-forming coccolithophore Emiliania huxleyi and its specific coccolithoviruses (E. huxleyi virus [EhV]). We show that EhV induces profound transcriptome remodeling targeted toward fatty acid synthesis to support viral assembly. A metabolic shift toward production of viral-derived sphingolipids was detected during infection and coincided with downregulation of host de novo sphingolipid genes and induction of the viral-encoded homologous pathway. The depletion of host-specific sterols during lytic infection and their detection in purified virions revealed their novel role in viral life cycle. We identify an essential function of the mevalonate-isoprenoid branch of sterol biosynthesis during infection and propose its downregulation as an antiviral mechanism. We demonstrate how viral replication depends on the hijacking of host lipid metabolism during the chemical "arms race" in the ocean. PMID:24920329

  4. Rewiring Host Lipid Metabolism by Large Viruses Determines the Fate of Emiliania huxleyi, a Bloom-Forming Alga in the Ocean[C][W][OPEN

    PubMed Central

    Rosenwasser, Shilo; Mausz, Michaela A.; Schatz, Daniella; Sheyn, Uri; Malitsky, Sergey; Aharoni, Asaph; Weinstock, Eyal; Tzfadia, Oren; Ben-Dor, Shifra; Feldmesser, Ester; Pohnert, Georg; Vardi, Assaf

    2014-01-01

    Marine viruses are major ecological and evolutionary drivers of microbial food webs regulating the fate of carbon in the ocean. We combined transcriptomic and metabolomic analyses to explore the cellular pathways mediating the interaction between the bloom-forming coccolithophore Emiliania huxleyi and its specific coccolithoviruses (E. huxleyi virus [EhV]). We show that EhV induces profound transcriptome remodeling targeted toward fatty acid synthesis to support viral assembly. A metabolic shift toward production of viral-derived sphingolipids was detected during infection and coincided with downregulation of host de novo sphingolipid genes and induction of the viral-encoded homologous pathway. The depletion of host-specific sterols during lytic infection and their detection in purified virions revealed their novel role in viral life cycle. We identify an essential function of the mevalonate-isoprenoid branch of sterol biosynthesis during infection and propose its downregulation as an antiviral mechanism. We demonstrate how viral replication depends on the hijacking of host lipid metabolism during the chemical “arms race” in the ocean. PMID:24920329

  5. Bioactive lipid coating of bone allografts directs engraftment and fate determination of bone marrow-derived cells in rat GFP chimeras.

    PubMed

    Das, Anusuya; Segar, Claire E; Chu, Yihsuan; Wang, Tiffany W; Lin, Yong; Yang, Chunxi; Du, Xeujun; Ogle, Roy C; Cui, Quanjun; Botchwey, Edward A

    2015-09-01

    Bone grafting procedures are performed to treat wounds incurred during wartime trauma, accidents, and tumor resections. Endogenous mechanisms of repair are often insufficient to ensure integration between host and donor bone and subsequent restoration of function. We investigated the role that bone marrow-derived cells play in bone regeneration and sought to increase their contributions by functionalizing bone allografts with bioactive lipid coatings. Polymer-coated allografts were used to locally deliver the immunomodulatory small molecule FTY720 in tibial defects created in rat bone marrow chimeras containing genetically-labeled bone marrow for monitoring cell origin and fate. Donor bone marrow contributed significantly to both myeloid and osteogenic cells in remodeling tissue surrounding allografts. FTY720 coatings altered the phenotype of immune cells two weeks post-injury, which was associated with increased vascularization and bone formation surrounding allografts. Consequently, degradable polymer coating strategies that deliver small molecule growth factors such as FTY720 represent a novel therapeutic strategy for harnessing endogenous bone marrow-derived progenitors and enhancing healing in load-bearing bone defects. PMID:26125501

  6. Immunoaffinity column clean-up prior to thin-layer chromatography for the determination of aflatoxins in various food matrices.

    PubMed

    Stroka, J; Van Otterdijk, R; Anklam, E

    2000-12-29

    A one-dimensional TLC method to determine aflatoxins (B1, B2, G1, G2) in various food matrices was elaborated which abstains fully on the use of chlorinated solvents. It implements an immunoaffinity clean-up step after extraction with methanol. The aflatoxins were quantified by densitometry. The method has shown to be rapid and efficient. In-house performance characteristics were established. The limit of quantification was found to be significantly lower than current regulatory limits for aflatoxin control outside and within the European Community. The obtained recovery and precision data gave a strong indication, that the method is likely to give satisfactory performance if tested in a future collaborative trial. PMID:11204238

  7. Validated determination of primulasaponins in primula root by a high-performance-thin-layer-chromatography densitometric approach.

    PubMed

    Coran, Silvia A; Mulas, Stefano

    2012-11-01

    A novel HPTLC-densitometric method was developed for separation and quantitation of primulasaponin I and II in different matrices. HPTLC silica gel 60 F254(S), 20 cm × 10 cm, plates with ethyl acetate:water:formic acid (5:1:1 v/v) as the mobile phase were used. Densitometric determinations were performed in reflectance mode at 540 nm after derivatization with vanillin reagent. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Primulasaponins were quantified in the range of 150-450 ng with RSD of repeatability and intermediate precision between 0.8 and 1.4% and accuracy within the acceptance limits. The method was tested on commercial herbal medicinal preparations claiming to contain primula root extract.

  8. Long and Short Lipid Molecules Experience the Same Interleaflet Drag in Lipid Bilayers

    NASA Astrophysics Data System (ADS)

    Horner, Andreas; Akimov, Sergey A.; Pohl, Peter

    2013-06-01

    Membrane interleaflet viscosity ηe affects tether formation, phase separation into domains, cell shape changes, and budding. Contrary to the expected contribution to interleaflet coupling from interdigitation, the slide of lipid patches in opposing monolayers conferred the same value ηe≈3×109Jsm-4 for the friction experienced by the ends of both short and long chain fluorescent lipid analogues. Consistent with the weak dependence of the translational diffusion coefficient on lipid length, the in-layer viscosity was, albeit length dependent, much smaller than ηe.

  9. Analysis of meibum and tear lipids.

    PubMed

    Pucker, Andrew D; Nichols, Jason J

    2012-10-01

    The meibum is a lipid-rich secretion that is the primary component of the external layer of the tear film. The meibomian glands produce the meibum, and meibomian gland dysfunction can lead to degradation of the tear film. Such dysfunction can result in ocular irritation, inflammation, and clinical disease. Understanding this relationship is critical to preventing ocular disease; therefore, a search of peer-reviewed literature focusing on the collection, quantification, and analysis of normal and abnormal meibum and tear lipids was conducted. Numerous collection and quantification techniques are described, including their advantages and disadvantages. Studies indicate that the meibum and tear lipids consist of a large array of polar and nonpolar lipids; individual lipids or their classes can be correlated to pathology. Significant amounts of lipids are deposited on contact lenses, depending on the nature of their polymer chemistry. These findings taken together indicate that normal meibum and tear lipids are essential for normal ocular health. Additional studies are required to provide a better understanding of the meibum and tear film biomolecules so that more effective treatments for blepharitis, dry eye disease, and tear film-related contact lens complications can be devised.

  10. A new estimation of the total flavonoids in silkworm cocoon sericin layer through aglycone determination by hydrolysis-assisted extraction and HPLC-DAD analysis

    PubMed Central

    Zhao, Jin-Ge; Zhang, Yu-Qing

    2016-01-01

    Background Silk sericin and a few non-protein components isolated from the cocoon layer including two silk proteins in silkworm Bombyx mori has many bioactivities. The dietary sericin possess antinatural oxidation, anticancer, antihyperlipidemic, and antidiabetic activities. The non-protein components surrounding the sericin layer involve in wax, pigments mainly meaning flavonoids, sugars, and other impurities. However, very few investigations have reported the estimation of the total flavonoids derived from the cocoon layer. The flavonoids are commonly present in their glycosylated forms and mostly exist as quercetin glycosides in the sericin layers of silkworm cocoons. Objective The aim of this study was to find a more accurate method to estimate the level of the total flavonoids in silkworm cocoons. Design An efficient procedure of hydrolysis-assisted extraction (HAE) was first established to estimate the level of the total flavonoids through the determination of their aglycones, quercetin, and kaempferol. Then, a comparison was made between traditional colorimetric method and our method. In addition, the antioxidant activities of hydrolysis-assisted extract sample were determined. Results The average contents of quercetin and kaempferol were 1.98 and 0.42 mg/g in Daizo cocoon. Their recoveries were 99.56 and 99.17%. The total sum of quercetin and kaempferol was detected to be 2.40±0.07 mg/g by HAE-HPLC, while the total flavonoids (2.59±0.48 mg/g) estimated by the traditional colorimetric method were only equivalent to 1.28±0.04 mg/g of quercetin. The HAE sample also exhibits that IC50 values of scavenging ability of diphenyl picryl hydrazinyl (DPPH) radical and hydroxyl radical (HO·) are 243.63 µg/mL and 4.89 mg/mL, respectively. Conclusions These results show that the HAE-HPLC method is specificity of cocoon and far superior to the colorimetric method. Therefore, this study has profound significance for the comprehensive utilization of silkworm cocoon and

  11. Molecular Dynamics Simulation Study of Permeation of Molecules through Skin Lipid Bilayer.

    PubMed

    Gupta, Rakesh; Sridhar, D B; Rai, Beena

    2016-09-01

    Stratum Corneum (SC), the outermost layer of skin, is mainly responsible for skin's barrier function. The complex lipid matrix of SC determines these barrier properties. In this study, the lipid matrix is modeled as an equimolar mixture of ceramide (CER), cholesterol (CHOL), and free fatty acid (FFA). The permeation of water, oxygen, ethanol, acetic acid, urea, butanol, benzene, dimethyl sulfoxide (DMSO), toluene, phenol, styrene, and ethylbenzene across this layer is studied using a constrained MD simulations technique. Several long constrained simulations are performed at a skin temperature of 310 K under NPT conditions. The free energy profiles and diffusion coefficients along the bilayer normal have been calculated for each molecule. Permeability coefficients are also calculated and compared with experimental data. The main resistance for the permeation of hydrophilic and hydrophobic permeants has been found to be in the interior of the lipid bilayer and near the lipid-water interface, respectively. The obtained permeability is found to be a few orders of magnitude higher than experimental values for hydrophilic molecules while for hydrophobic molecules more discrepancy was observed. Overall, the qualitative ranking is consistent with the experiments. PMID:27518707

  12. Waxes: A Forgotten Topic in Lipid Teaching.

    ERIC Educational Resources Information Center

    Dominguez, Eva; Heredia, Antonio

    1998-01-01

    Reviews the biological importance of the lipids categorized as waxes and describes some of the organic chemistry of these compounds. Presents a short laboratory exercise on the extraction of plant waxes and their analysis by thin layer chromatography. (Author/CCM)

  13. Isolation of Cellular Lipid Droplets: Two Purification Techniques Starting from Yeast Cells and Human Placentas

    PubMed Central

    Dalhaimer, Paul

    2014-01-01

    Lipid droplets are dynamic organelles that can be found in most eukaryotic and certain prokaryotic cells. Structurally, the droplets consist of a core of neutral lipids surrounded by a phospholipid monolayer. One of the most useful techniques in determining the cellular roles of droplets has been proteomic identification of bound proteins, which can be isolated along with the droplets. Here, two methods are described to isolate lipid droplets and their bound proteins from two wide-ranging eukaryotes: fission yeast and human placental villous cells. Although both techniques have differences, the main method - density gradient centrifugation - is shared by both preparations. This shows the wide applicability of the presented droplet isolation techniques. In the first protocol, yeast cells are converted into spheroplasts by enzymatic digestion of their cell walls. The resulting spheroplasts are then gently lysed in a loose-fitting homogenizer. Ficoll is added to the lysate to provide a density gradient, and the mixture is centrifuged three times. After the first spin, the lipid droplets are localized to the white-colored floating layer of the centrifuge tubes along with the endoplasmic reticulum (ER), the plasma membrane, and vacuoles. Two subsequent spins are used to remove these other three organelles. The result is a layer that has only droplets and bound proteins. In the second protocol, placental villous cells are isolated from human term placentas by enzymatic digestion with trypsin and DNase I. The cells are homogenized in a loose-fitting homogenizer. Low-speed and medium-speed centrifugation steps are used to remove unbroken cells, cellular debris, nuclei, and mitochondria. Sucrose is added to the homogenate to provide a density gradient and the mixture is centrifuged to separate the lipid droplets from the other cellular fractions. The purity of the lipid droplets in both protocols is confirmed by Western Blot analysis. The droplet fractions from both preps

  14. Lipid peroxidation is increased in tears from the elderly.

    PubMed

    Benlloch-Navarro, Soledad; Franco, Ilenia; Sánchez-Vallejo, Violeta; Silvestre, Dolores; Romero, Francisco Javier; Miranda, María

    2013-10-01

    We describe a procedure in which tears, obtained from Schirmer strips, are used to measure a marker of lipid peroxidation, malondialdehyde (MDA). We also compared the levels of proteins and MDA in tears from two groups of people: young adults (18-30 years old) and elderly adults (65-85 years old), because the data related to total protein concentration of human tears vary widely and because the majority of people over the age of 65 experience some symptoms of dry eyes and this condition has been recognized as an oxidative stress-induced disease. Our results show a significant difference in the protein concentration of the tears taken from the two age categories, younger adults (18-30 years old) and older adults (65-85 years old). Herein, we report for the first time an increase in MDA concentrations determined by HPLC in human tears based on age. It is possible that alterations in the tear lipid layer may lead to an increase in lipid peroxidation. Further studies are needed to understand the nature and function of tear film and stability in order to obtain new methods to analyze tears in patients with different diseases. In this sense, it would be interesting to compare MDA concentration in tears from control subjects and from people with meibomian gland dysfunction.

  15. Ex-situ gas diffusion layer intrusion effect determination of polymer electrolyte membrane fuel cell flow fields

    NASA Astrophysics Data System (ADS)

    Haase, S.; Rauber, M.

    2015-09-01

    In automotive PEM fuel cell systems, one of the most important targets is to reduce the parasitic power of balance of plant components, e.g. the air supply. This can be achieved for example by decreasing air stoichiometry. However, this could lead to bad flow sharing in the fuel cell stack. Therefore the fluid distribution in the flow field has to be evaluated, understood and optimized. This work evaluates the effect of GDL intrusion on the pressure drop via ex-situ determination of GDL intrusion using CFD simulation. The intruded GDL geometries, evaluated by an optical microscope with 200 times enlargement, are transferred to pressure drop behaviors by a numerical CFD model. These results are compared to the results of the differential pressure method of mapping the pressure distribution, described in [43]. The intrusion of the GDL leads to homogeneous flow distribution up to clamping pressures of 2.5 MPa. The inhomogeneous intrusion, induced by cracked fibers that extend into the channel, dominates the flow at higher clamping pressures and leads to the exponential increase in pressure drop in the differential pressure method. For clamping pressures used in typical fuel cell applications, the results of both methods show homogeneous flow through the channels.

  16. Validation of a Thin-Layer Chromatography for the Determination of Hydrocortisone Acetate and Lidocaine in a Pharmaceutical Preparation

    PubMed Central

    Dołowy, Małgorzata; Kulpińska-Kucia, Katarzyna; Pyka, Alina

    2014-01-01

    A new specific, precise, accurate, and robust TLC-densitometry has been developed for the simultaneous determination of hydrocortisone acetate and lidocaine hydrochloride in combined pharmaceutical formulation. The chromatographic analysis was carried out using a mobile phase consisting of chloroform + acetone + ammonia (25%) in volume composition 8 : 2 : 0.1 and silica gel 60F254 plates. Densitometric detection was performed in UV at wavelengths 200 nm and 250 nm, respectively, for lidocaine hydrochloride and hydrocortisone acetate. The validation of the proposed method was performed in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, and robustness. The applied TLC procedure is linear in hydrocortisone acetate concentration range of 3.75 ÷ 12.50 μg·spot−1, and from 1.00 ÷ 2.50 μg·spot−1 for lidocaine hydrochloride. The developed method was found to be accurate (the value of the coefficient of variation CV [%] is less than 3%), precise (CV [%] is less than 2%), specific, and robust. LOQ of hydrocortisone acetate is 0.198 μg·spot−1 and LOD is 0.066 μg·spot−1. LOQ and LOD values for lidocaine hydrochloride are 0.270 and 0.090 μg·spot−1, respectively. The assay value of both bioactive substances is consistent with the limits recommended by Pharmacopoeia. PMID:24526880

  17. Quantitative Determination of Flavonoids and Chlorogenic Acid in the Leaves of Arbutus unedo L. Using Thin Layer Chromatography.

    PubMed

    Maleš, Zeljan; Sarić, Darija; Bojić, Mirza

    2013-01-01

    The plant species Arbutus unedo shows numerous beneficial pharmacological effects (antiseptic, antidiabetic, antidiarrheal, astringent, depurative, antioxidant, antihypertensive, antithrombotic, and anti-inflammatory). For the medicinal use, standardization of extracts is a necessity, as different compounds are responsible for different biological activities. In this paper, we analyze monthly changes in the content of quercitrin, isoquercitrin, hyperoside, and chlorogenic acid. Methanolic extracts of the leaves are analyzed by HPTLC for the identification and quantification of individual polyphenol, and DPPH test is used to determine antioxidant activity. Based on the results obtained, the leaves should be collected in January to obtain the highest concentrations of hyperoside and quercitrin (0.35 mg/g and 1.94 mg/g, resp.), in June, July, and October for chlorogenic acid (1.45-1.46 mg/g), and for the fraction of quercitrin and isoquercitrin in November (1.98 mg/g and 0.33 mg/g, resp.). Optimal months for the collection of leaves with the maximum recovery of individual polyphenol suggested in this work could direct the pharmacological usage of the polyvalent herbal drugs. PMID:23984189

  18. Caenorhabditis elegans as an alternative in vivo model to determine oral uptake, nanotoxicity, and efficacy of melatonin-loaded lipid-core nanocapsules on paraquat damage

    PubMed Central

    Charão, Mariele Feiffer; Souto, Caroline; Brucker, Natália; Barth, Anelise; Jornada, Denise S; Fagundez, Daiandra; Ávila, Daiana Silva; Eifler-Lima, Vera L; Guterres, Silvia S; Pohlmann, Adriana R; Garcia, Solange Cristina

    2015-01-01

    Caenorhabditis elegans is an alternative in vivo model that is being successfully used to assess the pharmacological and toxic effects of drugs. The exponential growth of nanotechnology requires the use of alternative in vivo models to assess the toxic effects of theses nanomaterials. The use of polymeric nanocapsules has shown promising results for drug delivery. Moreover, these formulations have not been used in cases of intoxication, such as in treatment of paraquat (PQ) poisoning. Thus, the use of drugs with properties improved by nanotechnology is a promising approach to overcome the toxic effects of PQ. This research aimed to evaluate the absorption of rhodamine B-labeled melatonin (Mel)-loaded lipid-core nanocapsules (LNC) by C. elegans, the application of this model in nanotoxicology, and the protection of Mel-LNC against PQ damage. The formulations were prepared by self-assembly and characterized by particle sizing, zeta potential, drug content, and encapsulation efficiency. The results demonstrated that the formulations had narrow size distributions. Rhodamine B-labeled Mel-LNC were orally absorbed and distributed in the worms. The toxicity assessment of LNC showed a lethal dose 50% near the highest dose tested, indicating low toxicity of the nanocapsules. Moreover, pretreatment with Mel-LNC significantly increased the survival rate, reduced the reactive oxygen species, and maintained the development in C. elegans exposed to PQ compared to those worms that were either untreated or pretreated with free Mel. These results demonstrated for the first time the uptake and distribution of Mel-LNC by a nematode, and indicate that while LNC is not toxic, Mel-LNC prevents the effects of PQ poisoning. Thus, C. elegans may be an interesting alternative model to test the nanocapsules toxicity and efficacy. PMID:26300641

  19. Determination of toxic α-dicarbonyl compounds, glyoxal, methylglyoxal, and diacetyl, released to the headspace of lipid commodities upon heat treatment.

    PubMed

    Jiang, Yaping; Hengel, Matt; Pan, Canping; Seiber, James N; Shibamoto, Takayuki

    2013-02-01

    Toxic α-dicarbonyl compounds, glyoxal, 2-methylglyoxal, and diacetyl, released from the headspace from butter, margarine, safflower oil, beef fat, and cheese heated at 100 and 200 °C were analyzed by gas chromatography as quinoxaline derivatives. Total amounts of α-dicarbonyl compounds ranged from 40.5 ng/g (butter) to 331.2 ng/g (beef fat) at 100 °C and from 302.4 ng/g (safflower oil) to 4521.5 ng/g (margarine) at 200 °C. The total amount of α-dicarbonyl compounds increased approximately 55- and 15-fold in the headspace of heated butter and margarine, respectively, when the temperature was increased from 100 to 200 °C. However, only slight differences associated with temperature variation were observed in the cases of safflower oil and beef fat (1.3- and 1.1-fold, respectively). Diacetyl was found in the highest amounts among all samples, ranging from 13.9 ± 0.3 ng/g (butter) to 2835.7 ng/g (cheese) at 100 °C and from 112.5 ± 102 ng/g (safflower oil) to 2274.5 ± 442.6 ng/g (margarine) at 200 °C, followed by methylglyoxal, ranging from 13.0 ± 0.5 to 112.7 ± 10.1 ng/g (cheese) at 100 °C and from 34.7 ± 5.0 ng/g (safflower oil) to 1790 ± 372.3 ng/g (margarine) at 200 °C. Much less glyoxal formed, in amounts ranging from 13.6 ± 0.7 ng/g (butter) to 53.4 ± 11.2 ng/g (beef fat) at both temperatures. The amounts of α-dicarbonyl compounds released into the vapor phase from lipid commodities during heating were satisfactorily analyzed.

  20. Identification of furan fatty acids in the lipids of common carp (Cyprinus carpio L.).

    PubMed

    Chvalová, Daniela; Špička, Jiří

    2016-06-01

    Fatty acid (FA) composition was analyzed in muscle and gonad tissues of marketed common carp (Cyprinus carpio). The extracted lipids were separated into four fractions: polar lipids (PL), diacylglycerols, free fatty acids and triacylglycerols (TAG) using thin layer chromatography. FA content within the lipid fractions was determined by gas chromatography with flame ionization detector (GC/FID). The muscle lipids consisted primarily of TAG (96.9% of total FA), while PL were the major component of both male (67.6%) and female gonad (58.6%) lipids. Polyunsaturated fatty acids predominated in PL of all tissues (52.2-55.8% of total FA); monounsaturated fatty acids were the most abundant FA group in TAG of muscle (51.8%) and female gonads (47.8%) whereas high proportion of furan fatty acids (F-acids) (38.2%) was detected in TAG of male gonads. Eight F-acids were identified by gas chromatography-mass spectrometry (GC/MS) in male gonad samples, including less common 12,15-epoxy-13,14-dimethylnonadeca-12,14-dienoic acid with even-numbered alkyl moiety. PMID:26830577

  1. Lipid composition of multilamellar bodies secreted by Dictyostelium discoideum reveals their amoebal origin.

    PubMed

    Paquet, Valérie E; Lessire, René; Domergue, Frédéric; Fouillen, Laetitia; Filion, Geneviève; Sedighi, Ahmadreza; Charette, Steve J

    2013-10-01

    When they are fed with bacteria, Dictyostelium discoideum amoebae produce and secrete multilamellar bodies (MLBs), which are composed of membranous material. It has been proposed that MLBs are a waste disposal system that allows D. discoideum to eliminate undigested bacterial remains. However, the real function of MLBs remains unknown. Determination of the biochemical composition of MLBs, especially lipids, represents a way to gain information about the role of these structures. To allow these analyses, a protocol involving various centrifugation procedures has been developed to purify secreted MLBs from amoeba-bacterium cocultures. The purity of the MLB preparation was confirmed by transmission electron microscopy and by immunofluorescence using H36, an antibody that binds to MLBs. The lipid and fatty acid compositions of pure MLBs were then analyzed by high-performance thin-layer chromatography (HPTLC) and gas chromatography (GC), respectively, and compared to those of amoebae as well as bacteria used as a food source. While the bacteria were devoid of phosphatidylcholine (PC) and phosphatidylinositol (PI), these two polar lipid species were major classes of lipids in MLBs and amoebae. Similarly, the fatty acid composition of MLBs and amoebae was characterized by the presence of polyunsaturated fatty acids, while cyclic fatty acids were found only in bacteria. These results strongly suggest that the lipids constituting the MLBs originate from the amoebal metabolism rather than from undigested bacterial membranes. This opens the possibility that MLBs, instead of being a waste disposal system, have unsuspected roles in D. discoideum physiology.

  2. Lipid Composition of Multilamellar Bodies Secreted by Dictyostelium discoideum Reveals Their Amoebal Origin

    PubMed Central

    Paquet, Valérie E.; Lessire, René; Domergue, Frédéric; Fouillen, Laetitia; Filion, Geneviève; Sedighi, Ahmadreza

    2013-01-01

    When they are fed with bacteria, Dictyostelium discoideum amoebae produce and secrete multilamellar bodies (MLBs), which are composed of membranous material. It has been proposed that MLBs are a waste disposal system that allows D. discoideum to eliminate undigested bacterial remains. However, the real function of MLBs remains unknown. Determination of the biochemical composition of MLBs, especially lipids, represents a way to gain information about the role of these structures. To allow these analyses, a protocol involving various centrifugation procedures has been developed to purify secreted MLBs from amoeba-bacterium cocultures. The purity of the MLB preparation was confirmed by transmission electron microscopy and by immunofluorescence using H36, an antibody that binds to MLBs. The lipid and fatty acid compositions of pure MLBs were then analyzed by high-performance thin-layer chromatography (HPTLC) and gas chromatography (GC), respectively, and compared to those of amoebae as well as bacteria used as a food source. While the bacteria were devoid of phosphatidylcholine (PC) and phosphatidylinositol (PI), these two polar lipid species were major classes of lipids in MLBs and amoebae. Similarly, the fatty acid composition of MLBs and amoebae was characterized by the presence of polyunsaturated fatty acids, while cyclic fatty acids were found only in bacteria. These results strongly suggest that the lipids constituting the MLBs originate from the amoebal metabolism rather than from undigested bacterial membranes. This opens the possibility that MLBs, instead of being a waste disposal system, have unsuspected roles in D. discoideum physiology. PMID:23748431

  3. Nutrients and neurodevelopment: lipids.

    PubMed

    González, Horacio F; Visentin, Silvana

    2016-10-01

    Nutrients, lipids in particular, make up the central nervous system structure and play major functional roles: they stimulate development, migration, and nerve cell differentiation. They are part of gray matter, white matter, nerve nuclei, and synaptogenesis. Breast milk contains lipids which are crucial for infant brain development. The lipid profile of breast milk was used as a guideline for the development of breast milk substitutes. However, to date, no substitute has matched it. Complementary feeding should include docosahexaenoic acid, arachidonic acid, other polyunsaturated fatty acids, saturated fatty acids, and complex lipids found in milk fat. The lipid composition of breast milk depends on maternal intake and nutritional status during pregnancy and breast-feeding. It has a great impact on development. Our goal is to review scientific literature regarding the role of lipids on infant brain development and the importance of breast milk lipid composition, maternal diet, and complementary feeding. PMID:27606648

  4. Nutrients and neurodevelopment: lipids.

    PubMed

    González, Horacio F; Visentin, Silvana

    2016-10-01

    Nutrients, lipids in particular, make up the central nervous system structure and play major functional roles: they stimulate development, migration, and nerve cell differentiation. They are part of gray matter, white matter, nerve nuclei, and synaptogenesis. Breast milk contains lipids which are crucial for infant brain development. The lipid profile of breast milk was used as a guideline for the development of breast milk substitutes. However, to date, no substitute has matched it. Complementary feeding should include docosahexaenoic acid, arachidonic acid, other polyunsaturated fatty acids, saturated fatty acids, and complex lipids found in milk fat. The lipid composition of breast milk depends on maternal intake and nutritional status during pregnancy and breast-feeding. It has a great impact on development. Our goal is to review scientific literature regarding the role of lipids on infant brain development and the importance of breast milk lipid composition, maternal diet, and complementary feeding.

  5. Determination of the thermal diffusivity of bulk and layered samples by time domain thermoreflectance: Interest of lateral heat diffusion investigation in nanoscale time range

    SciTech Connect

    Belliard, L. Charron, E.; Vincent, S.; Perrin, B.; Fournier, D.; Frétigny, C.

    2015-02-14

    We report on thermal investigations performed in a time resolved experimental scheme. The time domain thermoreflectance (TDTR) is applied in an unusual geometry where the pump and probe beams are not superimposed but focused and shifted. In this way, the determination of the in-plane thermal diffusivity is achieved from temperature snapshots at different time delays. In the first part, taking into account the specific generation process and the detection inherent to the time domain thermoreflectance approach, an analytical solution for the temperature field is obtained for bulk samples, and compared to experimental data. A comparison with the frequency domain thermoreflectance microscopy is also outlined. In Part II section, the lateral heat diffusion in a layered structure is investigated. The comparison of the heat diffusion spreading in case of a highly conductive layer deposited on an insulator substrate and the reverse situation are carefully studied. Finally, we show how the time dependence is efficient to probe and identify material thermal properties or thermal interfacial resistance.

  6. Determination of the thermal diffusivity of bulk and layered samples by time domain thermoreflectance: Interest of lateral heat diffusion investigation in nanoscale time range

    NASA Astrophysics Data System (ADS)

    Belliard, L.; Frétigny, C.; Charron, E.; Vincent, S.; Perrin, B.; Fournier, D.

    2015-02-01

    We report on thermal investigations performed in a time resolved experimental scheme. The time domain thermoreflectance (TDTR) is applied in an unusual geometry where the pump and probe beams are not superimposed but focused and shifted. In this way, the determination of the in-plane thermal diffusivity is achieved from temperature snapshots at different time delays. In the first part, taking into account the specific generation process and the detection inherent to the time domain thermoreflectance approach, an analytical solution for the temperature field is obtained for bulk samples, and compared to experimental data. A comparison with the frequency domain thermoreflectance microscopy is also outlined. In Part II section, the lateral heat diffusion in a layered structure is investigated. The comparison of the heat diffusion spreading in case of a highly conductive layer deposited on an insulator substrate and the reverse situation are carefully studied. Finally, we show how the time dependence is efficient to probe and identify material thermal properties or thermal interfacial resistance.

  7. Determination of Marginal Conditions for Thermoacoustic Oscillations in a Looped Tube by Evolution of an Initial Disturbance Based on the Boundary-Layer Theory

    NASA Astrophysics Data System (ADS)

    Shimizu, Dai; Sugimoto, Nobumasa

    2014-03-01

    Marginal conditions are determined for the onset of thermoacoustic oscillations of a gas in a looped tube with a so-called stack sandwiched by hot and cold heat exchangers on the basis of the boundary-layer theory so far developed. Given a couple of impulses applied initially to a quiescent gas, the evolution of an infinitesimally small disturbance is studied by solving an initial-value problem to the linearized equations. Marginal states correspond to those in which the initial disturbance neither decays nor grows with time. In the case studied by Ueda and Kato [J. Acoust. Soc. Am. 124, 851 (2008)], the marginal conditions are obtained in two cases of the temperature distribution in the thermal buffer tube and compared with their results. To identify the marginal curves, the porosity of the stack or one of the subsidiary parameters such as the wall thickness of the stack is required in addition to the squared ratio of the hydraulic radius to the typical thickness of the thermal diffusion layer in a flow passage of gas. It is found that the marginal conditions are in qualitative agreement with the experimental results and that the right branch of the marginal curve is obtainable down to such a considerably low temperature ratio that the theory may become invalid.

  8. Dissolvable layered double hydroxide coated magnetic nanoparticles for extraction followed by high performance liquid chromatography for the determination of phenolic acids in fruit juices.

    PubMed

    Saraji, Mohammad; Ghani, Milad

    2014-10-31

    A magnesium-aluminum layered double hydroxide coated on magnetic nanoparticles was synthesized and used as a sorbent to extract some phenolic acids including p-hydroxy benzoic acid, caffeic acid, syringic acid, p-coumaric acid and ferulic acid from fruit juices. After extraction, the elution step was performed through dissolving double hydroxide layers containing the analytes by changing the solution pH. The extracted phenolic acids were separated and quantified using high performance liquid chromatography-photodiode array detection. Experimental parameters such as sorbent amount, solution pH, desorption solvent volume and extraction time were studied and optimized. The linearity range of the method was between 2 and 500μgL(-1) with the determination coefficient (r(2)) higher than 0.991. Relative standard deviations for intra- and inter-day precision for the analytes at 100μgL(-1) were in the range of 4.3-9.2% and 4.9-8.6%, respectively. Batch-to-batch reproducibility at 100μgL(-1) concentration level was in the range of 7.8-11% (n=3). The limits of detection were between 0.44 and 1.3μgL(-1). Relative recoveries higher than 81% with RSDs in the range of 4.2-9.7% were obtained in the analysis of fruit juice samples. PMID:25260344

  9. Crystallization modifiers in lipid systems.

    PubMed

    Ribeiro, Ana Paula Badan; Masuchi, Monise Helen; Miyasaki, Eriksen Koji; Domingues, Maria Aliciane Fontenele; Stroppa, Valter Luís Zuliani; de Oliveira, Glazieli Marangoni; Kieckbusch, Theo Guenter

    2015-07-01

    Crystallization of fats is a determinant physical event affecting the structure and properties of fat-based products. The stability of these processed foods is regulated by changes in the physical state of fats and alterations in their crystallization behavior. Problems like polymorphic transitions, oil migration, fat bloom development, slow crystallization and formation of crystalline aggregates stand out. The change of the crystallization behavior of lipid systems has been a strategic issue for the processing of foods, aiming at taylor made products, reducing costs, improving quality, and increasing the applicability and stability of different industrial fats. In this connection, advances in understanding the complex mechanisms that govern fat crystallization led to the development of strategies in order to modulate the conventional processes of fat structuration, based on the use of crystallization modifiers. Different components have been evaluated, such as specific triacyglycerols, partial glycerides (monoacylglycerols and diacylglycerols), free fatty acids, phospholipids and emulsifiers. The knowledge and expertise on the influence of these specific additives or minor lipids on the crystallization behavior of fat systems represents a focus of current interest for the industrial processing of oils and fats. This article presents a comprehensive review on the use of crystallization modifiers in lipid systems, especially for palm oil, cocoa butter and general purpose fats, highlighting: i) the removal, addition or fractionation of minor lipids in fat bases; ii) the use of nucleating agents to modify the crystallization process; iii) control of crystallization in lipid bases by using emulsifiers. The addition of these components into lipid systems is discussed in relation to the phenomena of nucleation, crystal growth, morphology, thermal behavior and polymorphism, with the intention of providing the reader with a complete panorama of the associated mechanisms

  10. Membrane-spanning lipids for an uncompromised monitoring of membrane fusion and intermembrane lipid transfer

    PubMed Central

    Schwarzmann, Günter; Breiden, Bernadette; Sandhoff, Konrad

    2015-01-01

    A Förster resonance energy transfer-based fusion and transfer assay was developed to study, in model membranes, protein-mediated membrane fusion and intermembrane lipid transfer of fluorescent sphingolipid analogs. For this assay, it became necessary to apply labeled reporter molecules that are resistant to spontaneous as well as protein-mediated intermembrane transfer. The novelty of this assay is the use of nonextractable fluorescent membrane-spanning bipolar lipids. Starting from the tetraether lipid caldarchaeol, we synthesized fluorescent analogs with fluorophores at both polar ends. In addition, we synthesized radioactive glycosylated caldarchaeols. These labeled lipids were shown to stretch through bilayer membranes rather than to loop within a single lipid layer of liposomes. More important, the membrane-spanning lipids (MSLs) in contrast to phosphoglycerides proved to be nonextractable by proteins. We could show that the GM2 activator protein (GM2AP) is promiscuous with respect to glycero- and sphingolipid transfer. Saposin (Sap) B also transferred sphingolipids albeit with kinetics different from GM2AP. In addition, we could unambiguously show that the recombinant activator protein Sap C x His6 induced membrane fusion rather than intermembrane lipid transfer. These findings showed that these novel MSLs, in contrast with fluorescent phosphoglycerolipids, are well suited for an uncompromised monitoring of membrane fusion and intermembrane lipid transfer. PMID:26269359

  11. Lipid metabolic reprogramming in cancer cells

    PubMed Central

    Beloribi-Djefaflia, S; Vasseur, S; Guillaumond, F

    2016-01-01

    Many human diseases, including metabolic, immune and central nervous system disorders, as well as cancer, are the consequence of an alteration in lipid metabolic enzymes and their pathways. This illustrates the fundamental role played by lipids in maintaining membrane homeostasis and normal function in healthy cells. We reviewed the major lipid dysfunctions occurring during tumor development, as determined using systems biology approaches. In it, we provide detailed insight into the essential roles exerted by specific lipids in mediating intracellular oncogenic signaling, endoplasmic reticulum stress and bidirectional crosstalk between cells of the tumor microenvironment and cancer cells. Finally, we summarize the advances in ongoing research aimed at exploiting the dependency of cancer cells on lipids to abolish tumor progression. PMID:26807644

  12. Matrix-elimination with steam distillation for determination of short-chain fatty acids in hypersaline waters from pre-salt layer by ion-exclusion chromatography.

    PubMed

    Ferreira, Fernanda N; Carneiro, Manuel C; Vaitsman, Delmo S; Pontes, Fernanda V M; Monteiro, Maria Inês C; Silva, Lílian Irene D da; Neto, Arnaldo Alcover

    2012-02-01

    A method for determination of formic, acetic, propionic and butyric acids in hypersaline waters by ion-exclusion chromatography (IEC), using steam distillation to eliminate matrix-interference, was developed. The steam distillation variables such as type of solution to collect the distillate, distillation time and volume of the 50% v/v H₂SO₄ solution were optimized. The effect of the addition of NaCl different concentrations to the calibration standards on the carboxylic acid recovery was also investigated. Detection limits of 0.2, 0.5, 0.3 and 1.5 mg L⁻¹ were obtained for formic, acetic, propionic and butyric acids, respectively. Produced waters from petroleum reservoirs in the Brazilian pre-salt layer containing about 19% m/v of NaCl were analyzed. Good recoveries (99-108%) were obtained for all acids in spiked produced water samples.

  13. Nickel-aluminum layered double hydroxide as a nanosorbent for selective solid-phase extraction and spectrofluorometric determination of salicylic acid in pharmaceutical and biological samples.

    PubMed

    Abdolmohammad-Zadeh, H; Kohansal, S; Sadeghi, G H

    2011-04-15

    The nickel-aluminum layered double hydroxide (Ni-Al LDH) was synthesized by a simple co-precipitation method and used as a solid-phase extraction (SPE) sorbent for separation and pre-concentration of trace levels of salicylic acid (SA) from aqueous solutions. Extraction of analyte is based on the adsorption of salicylate ions on the Ni-Al (NO(3)(-)) LDH and/or their exchanging with LDH interlayer NO(3)(-) ions. The retained analyte on the LDH was stripped by 3 mol L(-1) NaOH solution and its concentration was subsequently determined spectrofluorometrically at λ(em)=400 nm with excitation at λ(ex)=270 nm. Various parameters affecting the extraction efficiency of SA on the Ni-Al (NO(3)(-)) LDH, such as pH, amount of nano-sorbent, sample loading flow rate, elution conditions, sample volume and matrix effects were investigated. In the optimum experimental conditions, the limit of detection (3s) and enrichment factor were 0.12 μg L(-1) and 40, respectively. The relative standard deviation (RSD) for six replicate determinations of 10 μg L(-1) SA was 2.3%. The calibration graph using the pre-concentration system was linear in the range of 0.3-45 μg L(-1) with a correlation coefficient of 0.9985. The optimized method was successfully applied to the determination of SA in blood serum, willow leaf and aspirin tablet.

  14. Ultratrace determination of arsenic in water samples by electrothermal atomic absorption spectrometry after pre-concentration with Mg-Al-Fe ternary layered double hydroxide nano-sorbent.

    PubMed

    Abdolmohammad-Zadeh, Hossein; Jouyban, Abolghasem; Amini, Roghayeh

    2013-11-15

    A selective solid phase extraction method, based on nano-structured Mg-Al-Fe(NO3(-)) ternary layered double hydroxide as a sorbent, is developed for the pre-concentration of ultra-trace levels of arsenic (As) prior to determination by electrothermal atomic absorption spectrometry. It is found that both As(III) and As(V) could be quantitatively retained on the sorbent within a wide pH range of 4-12. Accordingly, the presented method is applied to determination of total inorganic As in aqueous solutions. Maximum analytical signal of As is achieved when the pyrolysis and atomization temperatures are close to 900 °C and 2300 °C, respectively. Several variables affecting the extraction efficiency including pH, sample flow rate, amount of nano-sorbent, elution conditions and sample volume are optimized. Under the optimized conditions, the limit of detection (3Sb/m) and the relative standard deviation are 4.6 pg mL(-1) and 3.9%, respectively. The calibration graph is linear in the range of 15.0-650 pg mL(-1) with a correlation coefficient of 0.9979, sorption capacity and pre-concentration factor are 8.68 mg g(-1) and 300, respectively. The developed method is validated by the analysis of a standard reference material (SRM 1643e) and is successfully applied to the determination of ultra-trace amounts of As in different water samples.

  15. Functional One-Dimensional Lipid Bilayers on Carbon Nanotube Templates

    SciTech Connect

    Artyukhin, A; Shestakov, A; Harper, J; Bakajin, O; Stroeve, P; Noy, A

    2004-07-23

    We present one-dimensional (1-D) lipid bilayer structures that integrate carbon nanotubes with a key biological environment-phospholipid membrane. Our structures consist of lipid bilayers wrapped around carbon nanotubes modified with a hydrophilic polymer cushion layer. Despite high bilayer curvature, the lipid membrane maintains its fluidity and can sustain repeated damage-recovery cycles. We also present the first evidence of spontaneous insertion of pore-forming proteins into 1-D lipid bilayers. These structures could lead to the development of new classes of biosensors and bioelectronic devices.

  16. Effect of chemical permeation enhancers on stratum corneum barrier lipid organizational structure and interferon alpha permeability.

    PubMed

    Moghadam, Shadi H; Saliaj, Evi; Wettig, Shawn D; Dong, Chilbert; Ivanova, Marina V; Huzil, J Torin; Foldvari, Marianna

    2013-06-01

    The outermost layer of the skin, known as the stratum corneum (SC), is composed of dead corneocytes embedded in an intercellular lipid matrix consisting of ceramides, free fatty acids, and cholesterol. The high level of organization within this matrix protects the body by limiting the permeation of most compounds through the skin. While essential for its protective functions, the SC poses a significant barrier for the delivery of topically applied pharmaceutical agents. Chemical permeation enhancers (CPEs) can increase delivery of small drug compounds into the skin by interacting with the intercellular lipids through physical processes including extraction, fluidization, increased disorder, and phase separation. However, it is not clear whether these same mechanisms are involved in delivery of biotherapeutic macromolecules, such as proteins. Here we describe the effect of three categories of CPEs {solvents [ethanol, propylene glycol, diethylene glycol monoethyl ether (transcutol), oleic acid], terpenes [menthol, nerol, camphor, methyl salicylate], and surfactants [Tween 80, SDS, benzalkonium chloride, polyoxyl 40 hydrogenated castor oil (Cremophor RH40), didecyldimethylammonium bromide (DDAB), didecyltrimethylammonium bromide (DTAB)]} on the lipid organizational structure of human SC as determined by X-ray scattering studies. Small- and wide-angle X-ray scattering studies were conducted to correlate the degree of structural changes and hydrocarbon chain packing in SC lipids caused by these various classes of CPEs to the extent of permeation of interferon alpha-2b (IFNα), a 19 kDa protein drug, into human skin. With the exception of solvents, propylene glycol and ethanol, all classes of CPEs caused increased disordering of lamellar and lateral packing of lipids. We observed that the highest degree of SC lipid disordering was caused by surfactants (especially SDS, DDAB, and DTAB) followed by terpenes, such as nerol. Interestingly, in vitro skin permeation studies

  17. Structural investigation of the covalent and electrostatic binding of yeast cytochrome c to the surface of various ultrathin lipid multilayers using x-ray diffraction.

    PubMed Central

    Pachence, J M; Blasie, J K

    1991-01-01

    X-Ray diffraction was used to characterize the profile structures of ultrathin lipid multilayers having a bound surface layer of cytochrome c. The lipid multilayers were formed on an alkylated glass surface, using the Langmuir-Blodgett method. The ultrathin lipid multilayers of this study were: five monolayers of arachidic acid, four monolayers of arachidic acid with a surface monolayer of dimyristoyl phosphatidylserine, and four monolayers of arachidic acid acid with a surface monolayer of thioethyl stearate. Both the phosphatidylserine and the thioethyl stearate surfaces were found previously to covalently bind yeast cytochrome c, while the arachidic acid surface electrostatically binds yeast cytochrome c. Meridional x-ray diffraction data were collected from these lipid multilayer films with and without a bound yeast cytochrome c surface layer. A box refinement technique, previously shown to be effective in deriving the profile structures of ultrathin multilayer lipid films with and without electrostatically bound cytochrome c, was used to determine the multilayer electron density profiles. The surface monolayer of bound cytochrome c was readily apparent upon comparison of the multilayer electron density profiles for the various pairs of ultrathin multilayer films plus/minus cytochrome c for all cases. In addition, cytochrome c binding to the multilayer surface significantly perturbs the underlying lipid monolayers. PMID:1648415

  18. Lipid compositional changes in calves fed excess iodine.

    PubMed

    Jenkins, K J

    1990-09-01

    Calves were fed milk replacer containing .57, 10, or 200 ppm iodine (from ethylenediaminedihydriodide) to determine the effects of excess dietary iodine on composition of lipids in blood plasma, liver, and heart. High iodine intakes had no effect on plasma total lipids or lipid classes, but caused lipid class concentration changes in liver and heart. Both 10 and 200 ppm iodine increased concentration of liver phosphatidylethanolamine and heart phosphatidylcholine, cholesterol, and total lipids, and the 200 ppm intake also increased concentration of liver phosphatidylcholine, total lipids, and heart phosphatidylethanolamine. Both iodine treatments tended to increase all the other minor lipid classes in liver and heart as well. Both 10 and 200 ppm iodine treatments increased some of the n-3 polyunsaturated fatty acids in the major phospholipids of plasma, liver, and heart. For the preruminant calf, liver and heart may be more useful than blood plasma for indicating excess iodine effects on lipid metabolism.

  19. Importance of the hexagonal lipid phase in biological membrane organization

    PubMed Central

    Jouhet, Juliette

    2013-01-01

    Domains are present in every natural membrane. They are characterized by a distinctive protein and/or lipid composition. Their size is highly variable from the nano- to the micrometer scale. The domains confer specific properties to the membrane leading to original structure and function. The determinants leading to domain organization are therefore important but remain obscure. This review presents how the ability of lipids to organize into hexagonal II or lamellar phases can promote particular local structures within membranes. Since biological membranes are composed of a mixture of lipids, each with distinctive biophysical properties, lateral and transversal sorting of lipids can promote creation of domains inside the membrane through local modulation of the lipid phase. Lipid biophysical properties have been characterized for long based on in vitro analyses using non-natural lipid molecules; their re-examinations using natural lipids might open interesting perspectives on membrane architecture occurring in vivo in various cellular and physiological contexts. PMID:24348497

  20. Distribution of the total unsaturation in lipid components of plasma as a new differential diagnostic method in clinical analysis.

    PubMed

    Poznyak, T I; Kiseleva, E V; Turkina, T I

    1997-08-01

    Using ozonization and thin-layer chromatographic methods we determined the qualitative and quantitative correlation of unsaturation distribution (UD) in individual fractions of blood plasma lipids in children suffering from insulin-dependent diabetes mellitus (IDDM). The research was aimed at elucidation of biochemical criterion of the degree of metabolic disorders in children with IDDM and at development of methods for quantitative assessment of such disorders. Twenty children were examined during the compensation stage (group 1), and twelve during decompensation with ketoacidosis (group 2). The present investigation shows that in the case of insulin-dependent diabetes mellitus in children the total unsaturation distribution (TUD) in plasma lipid fractions were found to be decreased significantly compared to healthy controls. The pattern of TUD in plasma lipid fractions may serve as a new biochemical criterion for metabolic disorders and decompensation in IDDM.

  1. Lipids of mitochondria.

    PubMed

    Horvath, Susanne E; Daum, Günther

    2013-10-01

    A unique organelle for studying membrane biochemistry is the mitochondrion whose functionality depends on a coordinated supply of proteins and lipids. Mitochondria are capable of synthesizing several lipids autonomously such as phosphatidylglycerol, cardiolipin and in part phosphatidylethanolamine, phosphatidic acid and CDP-diacylglycerol. Other mitochondrial membrane lipids such as phosphatidylcholine, phosphatidylserine, phosphatidylinositol, sterols and sphingolipids have to be imported. The mitochondrial lipid composition, the biosynthesis and the import of mitochondrial lipids as well as the regulation of these processes will be main issues of this review article. Furthermore, interactions of lipids and mitochondrial proteins which are highly important for various mitochondrial processes will be discussed. Malfunction or loss of enzymes involved in mitochondrial phospholipid biosynthesis lead to dysfunction of cell respiration, affect the assembly and stability of the mitochondrial protein import machinery and cause abnormal mitochondrial morphology or even lethality. Molecular aspects of these processes as well as diseases related to defects in the formation of mitochondrial membranes will be described.

  2. Molecular dynamics simulations of unsaturated lipid bilayers

    NASA Astrophysics Data System (ADS)

    Rabinovich, Alexander L.; Balabaev, Nikolay K.

    2001-02-01

    Molecular dynamics simulations were carried out for bilayers of lipid molecules having stearic acid (C18:0) chain in position '3-D' (using the nomenclature of M. Sundaralingam, 1972) and fatty acid chain C18:0, C18:1(omega 9), C18:2(omega 6), C18:3(omega 3), C20:4(omega 6) or C22:6(omega 3) in position '2-D'. To investigate the properties of the bilayers two models were considered. In the first model, the simulation cells of the bilayers consisted of 96 phosphatidylcholine (PC) molecules and 2304 water molecules: 48 lipid molecules per layer and 24 H2O molecules per lipid. The water was modeled by explicit TIP3P water molecules. In the second model, the head group of the lipid molecules was treated as an effective sphere -- diacylglycerolipids (DGs) were considered, the interface of each monolayer was modeled by a flat surface; no water molecules were present explicitly. The bilayers consisted of 48 X 2 equals 96 glycerolipids arranged in a rectangular simulation cell. Various properties of the bilayers -- the C-H bond order parameter -SCH profiles of the hydrocarbon tails, the root-mean-square values of the positional fluctuations of the lipid chain carbons, mass density distributions of lipid molecules and water along the normals were investigated.

  3. Molecular dynamics simulations of unsaturated lipid bilayers

    NASA Astrophysics Data System (ADS)

    Rabinovich, Alexander L.; Balabaev, Nikolay K.

    2000-02-01

    Molecular dynamics simulations were carried out for bilayers of lipid molecules having stearic acid (C18:0) chain in position '3-D' (using the nomenclature of M. Sundaralingam, 1972) and fatty acid chain C18:0, C18:1(omega 9), C18:2(omega 6), C18:3(omega 3), C20:4(omega 6) or C22:6(omega 3) in position '2-D'. To investigate the properties of the bilayers two models were considered. In the first model, the simulation cells of the bilayers consisted of 96 phosphatidylcholine (PC) molecules and 2304 water molecules: 48 lipid molecules per layer and 24 H2O molecules per lipid. The water was modeled by explicit TIP3P water molecules. In the second model, the head group of the lipid molecules was treated as an effective sphere -- diacylglycerolipids (DGs) were considered, the interface of each monolayer was modeled by a flat surface; no water molecules were present explicitly. The bilayers consisted of 48 X 2 equals 96 glycerolipids arranged in a rectangular simulation cell. Various properties of the bilayers -- the C-H bond order parameter -SCH profiles of the hydrocarbon tails, the root-mean-square values of the positional fluctuations of the lipid chain carbons, mass density distributions of lipid molecules and water along the normals were investigated.

  4. Lipid accumulation in prosthetic vascular grafts. Experimental study.

    PubMed Central

    Chignier, E.; Guidollet, J.; Lhopital, C.; Louisot, P.; Eloy, R.

    1990-01-01

    The present study demonstrates that the endoprosthetic tissue, developed at the contact of Dacron and Gore-Tex vascular prostheses replacing the infrarenal aortae of healthy dogs, presents a particular lipidic pattern as compared with the adjacent intimal arterial layer. The modified lipidic pattern is characterized by a significant increase in the total amounts of cholesterol, phospholipids, and triglycerides, despite a normal lipidic plasma profile. Histochemical studies showed that lipid droplets are accumulated in the cytoplasm of deeply situated cells and in the extracellular matrix. These findings support the idea that lipids may be trapped within the pseudo-intima of synthetic vascular grafts, even in the absence of a major plasma lipid disorder, and contribute to the prosthesis failure. Images Figure 2 Figure 4 Figure 5 Figure 6 PMID:2399933

  5. Determination of alpha-solanine and alpha-chaconine in potatoes by high-performance thin-layer chromatography/densitometry.

    PubMed

    Bodart, P; Kabengera, C; Noirfalise, A; Hubert, P; Angenot, L

    2000-01-01

    A high-performance thin-layer chromatographic (HPTLC) method was used to determine the glycoalkaloids alpha-solanine and alpha-chaconine in potatoes. Alpha-solanine and alpha-chaconine are extracted from dehydrated potatoes with boiling methanol-acetic acid (95 + 5, v/v). The analytes are separated on a Silica Gel 60 F254 HPTLC plate by a saturated mixture of dichloromethane-methanol-water-concentrated ammonium hydroxide (70 + 30 + 4 + 0.4, v/v), which is used for vertical development of the plate up to a distance of 85 mm. For visualization, the plate is dipped 3 times into a modified Carr-Price reagent, 20% (w/v) antimony(III) chloride in acetic acid-dichloromethane (1 + 3, v/v), and subsequently heated on a hot plate at 105 degrees C for 5 min. The glycoalkaloids all appear as red chromatographic zones on a colorless background. Densitometric quantification is performed at 507 nm by reflectance scanning. After determination of the appropriate response function, the proposed method was validated. Good results with respect to linearity, accuracy, and precision were obtained in the concentration range studied.

  6. Microfluidic Platforms for on-chip Formulation and Small-Angle x-ray Analysis of the Phase Behavior of Lipid/Water Mixtures

    SciTech Connect

    Khvostichenko, Daria S.; Perry, Sarah L.; Kondrashkina, Elena; Guha, Sudipto; Brister, Keith; Kenis, Paul J.A.

    2012-03-27

    We present a microfluidic platform for on-chip formulation and X-ray analysis of lipidic mesophases formed upon mixing lipids and water. The platform is designed to study the effect of detergents on the phase behavior of lipid/water mixtures. The platform allows automated preparation of multiple samples of different composition from stock solutions and subsequent on-chip small-angle X-ray diffraction (SAXS) data collection. To ensure X-ray transparency of the platform we used thin layers of cyclic olefin copolymer (COC) and PDMS. The viability of the platform is demonstrated by mapping out a section of the phase diagram for lipid monoolein mixed with solutions of detergent {beta}-octylglucoside. The platform reported here is a viable alternative to the traditional method of establishing phase diagrams for lipid/solution mixtures. Compared to the conventional approach, a significantly smaller amount of sample is required for mapping phase diagrams of lipidic mesophases and samples of various compositions are prepared automatically. In ongoing work we are using these chips to rapidly determine the phase behavior of a range of lipids to establish their suitability for membrane protein crystallization, especially with respect to their sensitivity to detergent concentration.

  7. A quick colorimetric method for total lipid quantification in microalgae.

    PubMed

    Byreddy, Avinesh R; Gupta, Adarsha; Barrow, Colin J; Puri, Munish

    2016-06-01

    Discovering microalgae with high lipid productivity are among the key milestones for achieving sustainable biodiesel production. Current methods of lipid quantification are time intensive and costly. A rapid colorimetric method based on sulfo-phospho-vanillin (SPV) reaction was developed for the quantification of microbial lipids to facilitate screening for lipid producing microalgae. This method was successfully tested on marine thraustochytrid strains and vegetable oils. The colorimetric method results correlated well with gravimetric method estimates. The new method was less time consuming than gravimetric analysis and is quantitative for lipid determination, even in the presence of carbohydrates, proteins and glycerol. PMID:27050419

  8. Modelling encapsulation of gold and silver nanoparticles inside lipid nanotubes

    NASA Astrophysics Data System (ADS)

    Baowan, Duangkamon; Thamwattana, Ngamta

    2014-02-01

    Lipid nanotubes are of particular interest for use as a template to create various one-dimensional nanostructures and as a carrier for drug and gene delivery. Understanding the encapsulation process is therefore crucial for such development. This paper models the interactions between lipid nanotubes and spheres of gold and silver nanoparticles and determines the critical dimension of lipid nanotubes that maximises the interaction with the nanoparticles. Our results confirm the acceptance of gold and silver nanoparticles inside lipid nanotubes. Further, we find that the lipid nanotube of radius approximately 10.23 nm is most favourable to encapsulate both types of nanoparticles.

  9. Improved characterization of EV preparations based on protein to lipid ratio and lipid properties.

    PubMed

    Osteikoetxea, Xabier; Balogh, Andrea; Szabó-Taylor, Katalin; Németh, Andrea; Szabó, Tamás Géza; Pálóczi, Krisztina; Sódar, Barbara; Kittel, Ágnes; György, Bence; Pállinger, Éva; Matkó, János; Buzás, Edit Irén

    2015-01-01

    In recent years the study of extracellular vesicles has gathered much scientific and clinical interest. As the field is expanding, it is becoming clear that better methods for characterization and quantification of extracellular vesicles as well as better standards to compare studies are warranted. The goal of the present work was to find improved parameters to characterize extracellular vesicle preparations. Here we introduce a simple 96 well plate-based total lipid assay for determination of lipid content and protein to lipid ratios of extracellular vesicle preparations from various myeloid and lymphoid cell lines as well as blood plasma. These preparations included apoptotic bodies, microvesicles/microparticles, and exosomes isolated by size-based fractionation. We also investigated lipid bilayer order of extracellular vesicle subpopulations using Di-4-ANEPPDHQ lipid probe, and lipid composition using affinity reagents to clustered cholesterol (monoclonal anti-cholesterol antibody) and ganglioside GM1 (cholera toxin subunit B). We have consistently found different protein to lipid ratios characteristic for the investigated extracellular vesicle subpopulations which were substantially altered in the case of vesicular damage or protein contamination. Spectral ratiometric imaging and flow cytometric analysis also revealed marked differences between the various vesicle populations in their lipid order and their clustered membrane cholesterol and GM1 content. Our study introduces for the first time a simple and readily available lipid assay to complement the widely used protein assays in order to better characterize extracellular vesicle preparations. Besides differentiating extracellular vesicle subpopulations, the novel parameters introduced in this work (protein to lipid ratio, lipid bilayer order, and lipid composition), may prove useful for quality control of extracellular vesicle related basic and clinical studies.

  10. Lipid Droplets And Cellular Lipid Metabolism

    PubMed Central

    Walther, Tobias C.; Farese, Robert V.

    2013-01-01

    Among organelles, lipid droplets (LDs) uniquely constitute a hydrophobic phase in the aqueous environment of the cytosol. Their hydrophobic core of neutral lipids stores metabolic energy and membrane components, making LDs hubs for lipid metabolism. In addition, LDs are implicated in a number of other cellular functions, ranging from protein storage and degradation to viral replication. These processes are functionally linked to many physiological and pathological conditions, including obesity and related metabolic diseases. Despite their important functions and nearly ubiquitous presence in cells, many aspects of LD biology are unknown. In the past few years, the pace of LD investigation has increased, providing new insights. Here, we review the current knowledge of LD cell biology and its translation to physiology. PMID:22524315

  11. Lipid exchange between membranes.

    PubMed Central

    Jähnig, F

    1984-01-01

    The exchange of lipid molecules between vesicle bilayers in water and a monolayer forming at the water surface was investigated theoretically within the framework of thermodynamics. The total number of exchanged molecules was found to depend on the bilayer curvature as expressed by the vesicle radius and on the boundary condition for exchange, i.e., whether during exchange the radius or the packing density of the vesicles remains constant. The boundary condition is determined by the rate of flip-flop within the bilayer relative to the rate of exchange between bi- and monolayer. If flip-flop is fast, exchange is independent of the vesicle radius; if flip-flop is slow, exchange increases with the vesicle radius. Available experimental results agree with the detailed form of this dependence. When the theory was extended to exchange between two bilayers of different curvature, the direction of exchange was also determined by the curvatures and the boundary conditions for exchange. Due to the dependence of the boundary conditions on flip-flop and, consequently, on membrane fluidity, exchange between membranes may partially be regulated by membrane fluidity. PMID:6518251

  12. The antagonist activity of lipid IVa on the stimulation by lipid A of TNF-alpha production from canine blood mononuclear cells.

    PubMed

    Takasawa, Kenji; Kano, Rui; Maruyama, Haruhiko; Hasegawa, Atsuhiko; Kamata, Hiroshi

    2011-09-15

    Lipid A, the active component of lipopolysaccharide (LPS), exists in the outer membrane of Gram-negative bacteria and binds to the Toll-like receptor 4 (TLR4) and MD-2 complex. On the other hand, the synthetic precursor of Escherichia coli lipid A, tetraacylated lipid IVa, is an agonist for TLR4 and MD-2 complex in murine, equine and feline cells but is an antagonist for lipid A in human cells. The aim of the study was to examine the function of canine Toll-like receptor 4 (TLR4) and MD-2 complex on canine blood mononuclear cells (BMC), by analyzing lipid A- or lipid IVa-induction of TNF-α production from these cells in order to understand canine innate immune system. After 5-h culture of canine BMC with lipid A (lipid A culture) or lipid IVa (lipid IVa culture), the TNF-α, as determined by ELISA, had increased in the supernatants of the lipid A cultures in a dose-dependent manner, whereas the TNF-α was undetectable in supernatant of lipid IVa-treated cultures. The TNF-α was statistically significantly different between the lipid A and lipid IVa cultures (100 and 1000 ng/ml). TNF-α production from canine BMC was inhibited, in a lipid IVa-dose-dependent manner, when the BMC were pre-cultured with lipid IVa for 60 min and then cultured with lipid A for 5h, while in control BMC cultures production if TNF-α was unchanged. These results indicate that the TNF-α production stimulated by lipid A was competed out by pre-exposing the BMC to lipid IVa. Thus, lipid A is an agonist for TNF-α production in canine BMC, whereas lipid IVa appears to be an antagonist against this lipid A stimulation of canine BMC.

  13. Results of tests on a Rockwell International space shuttle orbiter (-139 configuration) 0.0175-scale model (no. 29-0) in AEDC tunnel B to determine boundary layer characteristics

    NASA Technical Reports Server (NTRS)

    Quan, M.

    1975-01-01

    Results of wind tunnel tests were conducted to determine boundary layer characteristics on the lower surface of a space shuttle orbiter. Total pressure and temperature profile data at various model stations were obtained using a movable, four-degree-of-freedom probe mechanism and static pressure taps on the model surface. During a typical run, the probe was located over a preselected model location, then driven down through the bondary layer until contact was made with the model surface.

  14. Skin surface lipids of the domestic chicken, and neutral lipid standards as stimuli for the penetration response of Austrobilharzia variglandis cercariae.

    PubMed

    Zibulewsky, J; Fried, B; Bacha, W J

    1982-10-01

    Lipids were extracted from the skin of 2-wk-old domestic chickens using sterile cotton gauze dampened with chloroform:methanol (2:1). Preparative thin-layer chromatography separated the skin lipids into six major fractions: phospholipids, free sterols, free fatty acids, triglycerides, methyl esters, sterol esters. The penetration response of the marine avian schistosome cercaria, Austrobilharzia variglandis, to chicken skin lipid fractions, and to neutral lipid standards, was tested by coating lipids on agar in a Petri dish containing a seawater overlay. All neutral lipids tested produced significantly greater penetration responses than the chloroform control. The phospholipid skin fraction killed cercariae. Lipid from whole chicken skin produced the greatest penetration response, followed by free fatty acids and free sterol skin fractions. Of the standards tested, the whole neutral lipid standard, containing cholesterol, oleic acid, triolein, methyl oleate, and cholesteryl oleate, produced the greatest response, followed by the cholesterol standard and the oleic acid standard. PMID:7131196

  15. Numerical solution for 5-layer laminate technique to determine saturation solubility of a drug in a thin film of pressure sensitive adhesive.

    PubMed

    Bänsch, Eberhard; Reismann, Simone; Lee, Geoffrey

    2014-08-01

    A numerical solution of the one-dimensional diffusion equation is presented to describe the 5-layer laminate technique for estimating the saturation solubility of a drug in a thin polymer film. The boundary and initial conditions encompass a donor layer, a separating membrane, and an acceptor layer. Alteration of the drug's partition coefficient between donor and separating membrane has little influence on drug accumulation with the acceptor. The diffusivity in the separating membrane should be high to promote a short experimental time to achieve saturation equilibrium in the acceptor layer. The essential parameter to give rapid equilibrium is the thickness of the acceptor polymer film. For values of diffusivity typical for drugs of molecular weight around 500 an acceptor layer thickness of 10 µm-20 µm is required to achieve equilibrium within less than 10 d. These simulations allow the selection of suitable experimental conditions to make the 5-layer laminate technique a viable method for routine use. PMID:23944998

  16. New Salting Out Stability-Indicating and Kinetic Thin Layer Chromatographic Method for Determination of Glimepiride and Metformin HCl Binary Mixture.

    PubMed

    Mohamed, Yahya Abduh Salim; Mohamed, Abdel Maaboud Ismail; Mohamed, Fardous Abdel-Fattah; Ahmed, Sameh Abdel-Raouf

    2015-10-01

    A simple, selective salting out and stability-indicating thin layer chromatographic (SOTLC) technique was developed for determination of two antidiabetic drugs; glimepiride and metformin HCl in pure and in tablets as a binary mixture. Separation was performed on silica gel 60 F254 plates using aqueous ammonium sulfate and acetonitrile (7:3, v/v) as a mobile phase. The Rf values were 0.26 ± 0.02 and 0.73 ± 0.02 for glimepiride and metformin HCl, respectively. The separated bands were scanned at λ 237 nm using CAMAG TLC scanner III. The proposed method focusing on study of all the factors that play important role in the mechanism of salting out process. The proposed method was validated according to ICH guidelines and complied with USP31-NF26 validation guidelines. The correlation coefficients of calibration curves were 0.996 and 0.997 for glimepiride and metformin HCl, respectively, in the concentration range of 60-1,400 ng/band for both drugs. The investigated drugs were also subjected to acidic, basic, oxidative and photo-degradation and kinetic study was carried out.

  17. Determination of the magnetic contribution to the heat capacity of cobalt oxide nanoparticles and the thermodynamic properties of the hydration layers.

    PubMed

    Spencer, E C; Ross, N L; Parker, S F; Woodfield, B F; Boerio-Goates, J; Smith, S J; Olsen, R E; Kolesnikov, A I; Navrotsky, A; Ma, C

    2011-05-25

    We present low temperature (11 K) inelastic neutron scattering (INS) data on four hydrated nanoparticle systems: 10 nm CoO·0.10H(2)O (1), 16 nm Co(3)O(4)·0.40H(2)O (2), 25 nm Co(3)O(4)·0.30H(2)O (3) and 40 nm Co(3)O(4)·0.026H(2)O (4). The vibrational densities of states were obtained for all samples and from these the isochoric heat capacity and vibrational energy for the hydration layers confined to the surfaces of these nanoparticle systems have been elucidated. The results show that water on the surface of CoO nanoparticles is more tightly bound than water confined to the surface of Co(3)O(4), and this is reflected in the reduced heat capacity and vibrational entropy for water on CoO relative to water on Co(3)O(4) nanoparticles. This supports the trend, seen previously, for water to be more tightly bound in materials with higher surface energies. The INS spectra for the antiferromagnetic Co(3)O(4) particles (2-4) also show sharp and intense magnetic excitation peaks at 5 meV, and from this the magnetic contribution to the heat capacity of Co(3)O(4) nanoparticles has been calculated; this represents the first example of use of INS data for determining the magnetic contribution to the heat capacity of any magnetic nanoparticle system.

  18. Analysis of changes in hardness of a metal surface layer in areas of high stress and methods of determining residual life of parts for mining machines

    NASA Astrophysics Data System (ADS)

    Zvonarev, I. E.; Ivanov, S. L.

    2016-02-01

    The methodological bases for determining the energy resource of mechanical transmissions details for mining machines are considered. Based on the analysis of the accumulation of damage in metal gears, a method of estimating residual life of coarse-toothed wheels by periodically measuring the hardness of the surface layer of the teeth is justified. The regularities in change of hardness of coarse-tooth gear, conditioned by a change in metal strength properties that take into account the micro- and macromechanisms of plastic and elastic deformation, distortion of the metal crystal lattice with formation and movement of vacancies and dislocations. Experimental setup was built and the results of laboratory experiments are given related to the process of destruction of non-standard samples under different loads. Comparison of dimensions and hardness values of the sample allows concluding that a larger deformation corresponds to a greater increase in hardness, their limit value for the material being in the fracture zone. It is established that the detected changes in the local hardness occurs in areas of increased stresses above the limit of proportionality and the work of fracture forces attributed to dislocations density adjacent to the fracture plane expressed in terms of hardness increment is constant.

  19. New Salting Out Stability-Indicating and Kinetic Thin Layer Chromatographic Method for Determination of Glimepiride and Metformin HCl Binary Mixture.

    PubMed

    Mohamed, Yahya Abduh Salim; Mohamed, Abdel Maaboud Ismail; Mohamed, Fardous Abdel-Fattah; Ahmed, Sameh Abdel-Raouf

    2015-10-01

    A simple, selective salting out and stability-indicating thin layer chromatographic (SOTLC) technique was developed for determination of two antidiabetic drugs; glimepiride and metformin HCl in pure and in tablets as a binary mixture. Separation was performed on silica gel 60 F254 plates using aqueous ammonium sulfate and acetonitrile (7:3, v/v) as a mobile phase. The Rf values were 0.26 ± 0.02 and 0.73 ± 0.02 for glimepiride and metformin HCl, respectively. The separated bands were scanned at λ 237 nm using CAMAG TLC scanner III. The proposed method focusing on study of all the factors that play important role in the mechanism of salting out process. The proposed method was validated according to ICH guidelines and complied with USP31-NF26 validation guidelines. The correlation coefficients of calibration curves were 0.996 and 0.997 for glimepiride and metformin HCl, respectively, in the concentration range of 60-1,400 ng/band for both drugs. The investigated drugs were also subjected to acidic, basic, oxidative and photo-degradation and kinetic study was carried out. PMID:26006135

  20. Determination of the magnetic contribution to the heat capacity of cobalt oxide nanoparticles and the thermodynamic properties of the hydration layers

    SciTech Connect

    Spencer, Elinor; Ross, Dr. Nancy; Parker, Stewart F.; Woodfield, Brian; Boerio-Goates, Juliana; Smith, S. J.; Olsen, R. E.; Kolesnikov, Alexander I; Navrotsky, Alexandra; Ma, C

    2011-01-01

    We present low temperature (11 K) inelastic neutron scattering (INS) data on four hydrated nanoparticle systems: 10 nm CoO 0.10H2O (1), 16 nmCo3O4 0.40H2O (2), 25 nm Co3O4 0.30H2O (3) and 40 nmCo3O4 0.026H2O (4). The vibrational densities of states were obtained for all samples and from these the isochoric heat capacity and vibrational energy for the hydration layers confined to the surfaces of these nanoparticle systems have been elucidated. The results show that water on the surface of CoO nanoparticles is more tightly bound than water confined to the surface of Co3O4, and this is reflected in the reduced heat capacity and vibrational entropy for water on CoO relative to water on Co3O4 nanoparticles. This supports the trend, seen previously, for water to be more tightly bound in materials with higher surface energies. The INS spectra for the antiferromagnetic Co3O4 particles (2 4) also show sharp and intense magnetic excitation peaks at 5 meV, and from this the magnetic contribution to the heat capacity of Co3O4 nanoparticles has been calculated; this represents the first example of use of INS data for determining the magnetic contribution to the heat capacity of any magnetic nanoparticle system.

  1. Modelling uncertainty estimation for the determination of aflatoxin M(1) in milk by visual and densitometric thin-layer chromatography with immunoaffinity column clean-up.

    PubMed

    Carvalho, K L; Gonçalves, G A A; Lopes, A L; Santos, E A; Vargas, E A; Magalhães, W F

    2012-01-01

    The uncertainty of aflatoxin M(1) concentration in milk, determined by thin-layer chromatography (TLC) with visual and densitometric quantification of the fluorescence intensities of the spots, was estimated using the cause-and-effect approach proposed by ISO GUM (Guide to the expression of uncertainty in measurement) following its main four steps. The sources of uncertainties due to volume measurements, visual and densitometric TLC calibration curve, allowed range for recovery variation and intermediary precision to be taken into account in the uncertainty budget. For volume measurements the sources of uncertainties due to calibration, resolution, laboratory temperature variation and repeatability were considered. For the quantification by visual readings of the intensity of the aflatoxin M(1) in the TLC the uncertainty arising from resolution calibration curves was modelled based on the intervals of concentrations between pairs of the calibration standard solutions. The uncertainty of the densitometric TLC quantification arising from the calibration curve was obtained by weighted least square (WLS) regression. Finally, the repeatability uncertainty of the densitometric peak areas or of the visual readings for the test sample solutions was considered. For the test samples with aflatoxin M(1) concentration between 0.02 and 0.5 µg l(-1), the relative expanded uncertainties, with approximately 95% of coverage probability, obtained for visual TLC readings were between 60% and 130% of the values predicted by the Horwitz model. For the densitometric TLC determination they were about 20% lower. The main sources of uncertainties in both visual and densitometric TLC quantification were the intermediary precision, calibration curve and recovery. The main source of uncertainty in the calibration curve in the visual TLC analysis was due to the resolution of the visual readings, whereas in the densitometric analysis it was due to the peak areas of test sample solutions

  2. Proto-Organism Kinetics: Evolutionary Dynamics of Lipid Aggregates with Genes and Metabolism

    NASA Astrophysics Data System (ADS)

    Rasmussen, Steen; Chen, Liaohai; Stadler, Bärbel M. R.; Stadler, Peter F.

    2004-02-01

    A synthetic proto-organism could be self-assembled by integrating a lipid proto-container with a proto-metabolic subsystem and a proto-genetic subsystem. This three-component system can use energy and nutrients by means of either redox or photo-chemical reactions, evolve its proto-genome by means of template directed replication, and ultimately die. The evolutionary dynamics of the proto-organism depends crucially on the chemical kinetics of its sub-systems and on their interplay. In this work the template replication kinetics is investigated and it is found that the product inhibition inherent in the ligation-like replication process allows for coexistence of unrelated self-replicating proto-genes in the lipid surface layer. The combined catalytic effects from the proto-genes on the metabolic production rates determine the fate of the strain protocell.

  3. Incorporation of liquid lipid in lipid nanoparticles for ocular drug delivery enhancement

    NASA Astrophysics Data System (ADS)

    Shen, Jie; Sun, Minjie; Ping, Qineng; Ying, Zhi; Liu, Wen

    2010-01-01

    The present work investigates the effect of liquid lipid incorporation on the physicochemical properties and ocular drug delivery enhancement of nanostructured lipid carriers (NLCs) and attempts to elucidate in vitro and in vivo the potential of NLCs for ocular drug delivery. The CyA-loaded or fluorescein-marked nanocarriers composed of Precifac ATO 5 and Miglyol 840 (as liquid lipid) were prepared by melting-emulsion technology, and the physicochemical properties of nanocarriers were determined. The uptake of nanocarriers by human corneal epithelia cell lines (SDHCEC) and rabbit cornea was examined. Ex vivo fluorescence imaging was used to investigate the ocular distribution of nanocarriers. The in vitro cytotoxicity and in vivo acute tolerance were evaluated. The higher drug loading capacity and improved in vitro sustained drug release behavior of lipid nanoparticles was found with the incorporation of liquid lipid in lipid nanoparticles. The uptake of nanocarriers by the SDHCEC was increased with the increase in liquid lipid loading. The ex vivo fluorescence imaging of the ocular tissues indicated that the liquid lipid incorporation could improve the ocular retention and penetration of ocular therapeutics. No alternation was macroscopically observed in vivo after ocular surface exposure to nanocarriers. These results indicated that NLC was a biocompatible and potential nanocarrier for ocular drug delivery enhancement.

  4. Hydration in Lipid Monolayers: Correlation of Water Activity and Surface Pressure.

    PubMed

    Disalvo, E Anibal; Hollmann, Axel; Martini, M Florencia

    2015-01-01

    In order to give a physical meaning to each region of the membrane we define the interphase as the region in a lipid membrane corresponding to the polar head groups imbibed in water with net different properties than the hydrocarbon region and the water phase. The interphase region is analyzed under the scope of thermodynamics of surface and solutions based on the definition of Defay-Prigogine of an interphase and the derivation that it has in the understanding of membrane processeses in the context of biological response. In the view of this approach, the complete monolayer is considered as the lipid layer one molecule thick plus the bidimensional solution of the polar head groups inherent to it (the interphase region). Surface water activity appears as a common factor for the interaction of several aqueous soluble and surface active proteins with lipid membranes of different composition. Protein perturbation can be measured by changes in the surface pressure of lipid monolayers at different initial water surface activities. As predicted by solution chemistry, the increase of surface pressure is independent of the particle nature that dissolves. Therefore, membranes give a similar response in terms of the determined surface states given by water activity independent of the protein or peptide. PMID:26438267

  5. Integral hair lipid in human hair follicle.

    PubMed

    Lee, Won-Soo

    2011-12-01

    Integral hair lipid (IHL) is bound to the keratinized cell surface to make an environmentally resistant lipid envelope. It is mainly positioned on the hair cuticle and inner root sheath. IHL in the hair follicle may regard as hair barrier to be similar to the epidermal lipid layer functioning as skin barrier. Major constituents of IHL are fatty acid, phytosphingosine, ceramide in decreasing order. Minor constituents of IHL are cholesterol, cholesterol sulfate and cholesterol oleate. Cuticle or cortical cell surface in hair are abundant in fatty acids unlike the keratinized area of epidermis or sebaceous gland, and about 30-40% of such fatty acids are composed of 18-methyl-eicosanoic acid which is known to be bound to proteins by ester or thioester bond. Various factors including moisture, solvent, oxidative damage during bleaching or permanent waving affect IHL. Photochemical changes also can occur in IHL as well as in hair protein and hair pigment. Lipid metabolism is thought to play an essential role in lipid envelope of hair, but also involvement in hair development and function.

  6. Lipid peroxidation induced by shockwave lithotripsy.

    PubMed

    Cohen, T D; Durrani, A F; Brown, S A; Ferraro, R; Preminger, G M

    1998-06-01

    To determine the relation between high-energy shockwaves (HESW) and the presence of lipid peroxidation produces, juvenile pigs were subjected to shockwave lithotripsy (SWL). After lithotripsy, both treated and control kidneys were analyzed, along with urine samples collected before, during, and after SWL. Thiobarbituric acid-reactive substance (TBARS) and lipid-conjugated diene (CD) concentrations, used as markers for membrane lipid peroxidation, were determined in the kidney and urine samples. Significantly increased mean TBARS concentrations (146%) were associated with homogenates of lithotripsy-treated kidneys, 77.8 +/- 14.4 (SD) mmol/g v the controls, 31.4 +/- 14.9 mmol/g. Lithotripsy induction of lipid peroxidation products in the cortex, the gross damage site, and the respective medulla were also examined. In HESW-treated cortex samples, increased TBARS concentrations were seen--75.0 +/- 21.3 mmol/g--compared with untreated controls-- 45.2+/- 5.6 mmol/g--while increased CD concentrations (168%) were observed in the medulla of HESW-treated samples. No significant differences were observed in TBARS or CD concentrations in urine samples from control or treated kidneys, yet specific lipid hydroperperoxides were detected in the urine of HESW-treated kidneys. We conclude that HESW lithotripsy of swine kidneys is associated with increased lipid peroxidation products that may cause further cellular damage. Lipid peroxidation induced by SWL may be one of several mechanisms that lead to other potential bioeffects. Finally, analysis of specific lipid hydroperoxides in the urine of HESW-treated kidneys may serve as a noninvasive marker of renal injury after clinical SWL.

  7. Lipid-absorbing Polymers

    NASA Technical Reports Server (NTRS)

    Marsh, H. E., Jr.; Wallace, C. J.

    1973-01-01

    The removal of bile acids and cholesterol by polymeric absorption is discussed in terms of micelle-polymer interaction. The results obtained with a polymer composed of 75 parts PEO and 25 parts PB plus curing ingredients show an absorption of 305 to 309%, based on original polymer weight. Particle size effects on absorption rate are analyzed. It is concluded that crosslinked polyethylene oxide polymers will absorb water, crosslinked polybutadiene polymers will absorb lipids; neither polymer will absorb appreciable amounts of lipids from micellar solutions of lipids in water.

  8. Niacin, lipids, and heart disease.

    PubMed

    Malik, Shaista; Kashyap, Moti L

    2003-11-01

    Niacin is the most effective medication in current clinical use for increasing high-density lipoprotein (HDL) cholesterol. It has the broadest effect on the lipid profile, reducing all atherogenic apolipoprotein (apo) B and increasing all antiatherogenic apo AI-containing lipoproteins, resulting in significant reduction in atherosclerotic complications and total mortality in trials. Recent research indicates novel major target sites of action in the liver to 1) directly inhibit diacylglycerol acyltransferase 2 (DGAT2), explaining its effect on triglycerides and apo B lipoproteins, and 2) inhibit the HDL apo AI catabolism pathway, resulting in higher HDL levels. Such information may lead to new drug discovery and supply the rationale for combination with other lipid regulators that are known to have different mechanisms of action. Trial evidence shows that niacin is not only safe to use in persons with diabetes, but that its combination with 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG CoA) reductase inhibitors (statins) is also safe and effective. Recently, a new formulation of niacin has made it easier to tolerate and administer. Clinical trials are needed to determine whether niacin in combination with other lipid-modulating agents decreases the risk of cardiovascular events beyond the approximately 30% that has been noted with monotherapy.

  9. Using Vertical electrical sounding survey and refraction seismic survey for determining the geological layers depths, the structural features and assessment groundwater in Aqaba area in South Jordan.

    NASA Astrophysics Data System (ADS)

    Akawwi, Emad; Alzoubi, Abdallah; Ben Abraham, Zvi; Rahamn Abo Alades, Abdel; Alrzouq, Rami; Tiber, Gidon; Neimi, Tina

    2010-05-01

    The study area is the Aqaba region (Southern wadi Araba basin). Aqaba region area located at 87900 and 89000 North and 147000 and 158000 East (Palestine grid). Tectonically Aqaba area lies within the tectonic plate boundary along the Arabian and African plate slide. This plate boundary comprises numerous and shot fault segments. The main aims of this study are to assessing the groundwater potential and its quality, to explain the subsurface geological conditions and support the ongoing geological, environmental and hydrogeological studies. Therefore, it was anticipated that the results of the geophysical surveying will give many different important parameters as The subsurface geological features, thicknesses of the different lithological units, depth to the bed rocks and depth to the water table. The groundwater can apply an important role in ensuring sustainable water supply in the area. This study was carried out in order to assess groundwater condition, geological layers thicknesses and structural features in Aqaba area by using vertical electrical sounding (VES) surveys and refraction seismic techniques. There are three geoelectrical cross section were carried out at different sites by using the Schlumberger array. The first cross section indicated three layers of different resistivity. The second cross section indicated four layers of different resistivity. The third geoelectrical cross sections indicated three layers. The refraction seismic method also has been conducted in the same area as VES. About 12 refraction seismic profiles have been carried out in the study area. The length of the first profile was 745 m at the direction N-S. This profile indicated two different layers with a different velocities. The length of the second profile was 1320 m with E-W direction. This profile indicated two different layers. The length of the third profile was about 515 m with a direction SE-NW. It recognized two different layers with a different velocities. The fourth

  10. A method for determining fumonisin B1 in corn using immunoaffinity column clean-up and thin layer chromatography/densitometry.

    PubMed

    Preis, R A; Vargas, E A

    2000-06-01

    A method for determining fumonisin B1 (FB1) in corn was developed and the clean-up optimized in order to give an extract suitable for one-dimensional thin layer chromatographic (TLC) analysis. FB1 was extracted with a solution of methanol:water (80:20, v/v), purified through an immunoaffinity column and separated on a C18 reversed phase TLC plate. The FB1 was visualized with 0.1 mol/l sodium tetraborate, 0.40 mg/ml fluorescamine in acetonitrile and 0.01 mol/l boric acid:acetonitrile (2:3, v/v) for fluorescence detection, and quantified by densitometric analysis. Water, acetonitrile:water (1:1 v/v) and acetonitrile:water (4:1 v/v) were evaluated as TLC solvents for running both standards and samples together with derivatization procedures aimed at improving separation, resolution, sensitivity and linearity. The mean recovery for FB1 for spiked samples was found to be 85% and the linear equation of standard calibration curve by densitometric analysis gave an r2 value higher than 0.99. The maximum coefficient of variation for replicate analysis of spiked samples was 19%. The absolute amount of FB1 standard detectable on a TLC plate was 2 ng, giving a detection limit for the method of 0.1 mg/kg. The method has been shown to be robust in the application of FB1 monitoring in corn (214 samples) collected in different regions of the country. FB1 was detected in 99% of these samples in the range of 0.2 to 6 mg/kg. PMID:10932789

  11. Airborne determination of the temporo-spatial distribution of benzene, toluene, nitrogen oxides and ozone in the boundary layer across Greater London, UK

    NASA Astrophysics Data System (ADS)

    Shaw, M. D.; Lee, J. D.; Davison, B.; Vaughan, A.; Purvis, R. M.; Harvey, A.; Lewis, A. C.; Hewitt, C. N.

    2015-05-01

    Highly spatially resolved mixing ratios of benzene and toluene, nitrogen oxides (NOx) and ozone (O3) were measured in the atmospheric boundary layer above Greater London during the period 24 June to 9 July 2013 using a Dornier 228 aircraft. Toluene and benzene were determined in situ using a proton transfer reaction mass spectrometer (PTR-MS), NOx by dual-channel NOx chemiluminescence and O3 mixing ratios by UV absorption. Average mixing ratios observed over inner London at 360 ± 10 m a.g.l. were 0.20 ± 0.05, 0.28 ± 0.07, 13.2 ± 8.6, 21.0 ± 7.3 and 34.3 ± 15.2 ppbv for benzene, toluene, NO, NO2 and NOx respectively. Linear regression analysis between NO2, benzene and toluene mixing ratios yields a strong covariance, indicating that these compounds predominantly share the same or co-located sources within the city. Average mixing ratios measured at 360 ± 10 m a.g.l. over outer London were always lower than over inner London. Where traffic densities were highest, the toluene / benzene (T / B) concentration ratios were highest (average of 1.8 ± 0.5 ppbv ppbv-1), indicative of strong local sources. Daytime maxima in NOx, benzene and toluene mixing ratios were observed in the morning (~ 40 ppbv NOx, ~ 350 pptv toluene and ~ 200 pptv benzene) and in the mid-afternoon for ozone (~ 40 ppbv O3), all at 360 ± 10 m a.g.l.

  12. Airborne determination of the temporo-spatial distribution of benzene, toluene, nitrogen oxides and ozone in the boundary layer across Greater London, UK

    NASA Astrophysics Data System (ADS)

    Shaw, M. D.; Lee, J. D.; Davison, B.; Vaughan, A.; Purvis, R. M.; Lewis, A. C.; Hewitt, C. N.

    2014-10-01

    Highly spatially resolved mixing ratios of benzene and toluene, nitrogen oxides (NOx) and ozone (O3) were measured in the atmospheric boundary layer above Greater London during the period 24 June to 9 July 2013 using a Dornier 228 aircraft. Toluene and benzene were determined in-situ using a proton transfer reaction mass spectrometer (PTR-MS), NOx by dual channel NOx chemiluminescence and O3 mixing ratios by UV absorption. Average mixing ratios observed over inner London at 360 ± 10 m a.g.l. were 0.20 ± 0.05, 0.28 ± 0.07, 13.2 ± 8.6, 21.0 ± 7.3 and 34.3 ± 15.2 ppbv for benzene, toluene, NO, NO2 and NOx respectively. Linear regression analysis between NO2, benzene and toluene mixing ratios yielded a trimodal distribution indicating that these compounds predominantly share the same or co-located sources within the city and that a significant fraction of NOx is directly emitted as NO2. Average mixing ratios measured at 360 ± 10 m a.g.l. over outer London were always lower than over inner London. Where traffic densities were highest, the toluene / benzene (T / B) concentration ratios were highest (average of 1.8 ± 0.3 ppbv ppbv-1) indicative of strong local sources. Daytime maxima in NOx, benzene and toluene mixing ratios were observed in the morning (~40 ppbv NOx, ~350 pptv toluene and ~200 pptv benzene) and for ozone in the mid-afternoon (~40 ppbv O3) all at 360 ± 10 m a.g.l.

  13. Direct determination of the band offset in atomic layer deposited ZnO/hydrogenated amorphous silicon heterojunctions from X-ray photoelectron spectroscopy valence band spectra

    SciTech Connect

    Korte, L. Rößler, R.; Pettenkofer, C.

    2014-05-28

    The chemical composition and band alignment at the heterointerface between atomic layer deposition-grown zinc oxide (ZnO) and hydrogenated amorphous silicon (a-Si:H) is investigated using monochromatized X-ray photoelectron spectroscopy. A new approach for obtaining the valence band offset ΔE{sub V} is developed, which consists in fitting the valence band (VB) spectrum obtained for a-Si:H with a thin ZnO overlayer as the sum of experimentally obtained VB spectra of a bulk a-Si:H film and a thick ZnO film. This approach allows obtaining ΔE{sub V} = 2.71 ± 0.15 eV with a minimum of assumptions, and also yields information on the change in band bending of both substrate and ZnO film. The band offset results are compared to values obtained using the usual approach of comparing valence band edge-to-core level energy differences, ΔE{sub B,CL} − ΔE{sub B,VB}. Furthermore, a theoretical value for the VB offset is calculated from the concept of charge neutrality level line-up, using literature data for the charge neutrality levels and the experimentally determined ZnO/a-Si:H interface dipole. The thus obtained value of ΔE{sub V}{sup CNL} = 2.65 ± 0.3 eV agrees well with the experimental ΔE{sub V}.

  14. The development of a method for the qualitative and quantitative determination of petroleum hydrocarbon components using thin-layer chromatography with flame ionization detection.

    PubMed

    Wang, Shijie; Guo, Guanlin; Yan, Zengguang; Lu, Guilan; Wang, Qunhui; Li, Fasheng

    2010-01-15

    An analytical scheme to determine groups of petroleum hydrocarbon compounds in crude oil was developed and used for the qualitative and quantitative characterization of crude oil samples from the Shengli oilfield, the second largest oilfield in China. Crude oil samples were fractionated and analyzed by thin-layer chromatography with flame ionization detection (TLC-FID). Relative standard deviation (RSD) values for retention time, peak height and half peak width were less than 5.2% for all classes of compounds, based on nine independent replicates. The crude oil light fraction was further analyzed by GC-MS and the majority of identified compounds were methyl- or hydro-derivatives of long-chain hydrocarbons and aromatic compounds. The external standard method used in the present study can lower detection limits of petroleum hydrocarbon compound classes to 20.0 mg L(-1), and the crude oil concentration in the range of 30 and 35,000 mg L(-1) has a high linear correlation (r(2)>0.97, P<0.05) with peak area. A comparison between elution chromatography (EC) and TLC-FID regarding the recovery of petroleum hydrocarbon compounds was carried out with aged crude oil contaminated soils of 50, 80, 200 and 300 mg g(-1). The tested TLC-FID method showed a 10% higher recovery for total extractable materials than the reference EC method. The calibration factor was fraction-dependent and varied with the recovery rate of TLC/EC. Regarding the tested extraction procedures, accelerated solvent extraction (ASE) had a higher extraction efficiency for crude oil contaminated soils than Soxhlet and ultrasonic extractions.

  15. Determination of Sudan I in paprika powder by molecularly imprinted polymers-thin layer chromatography-surface enhanced Raman spectroscopic biosensor.

    PubMed

    Gao, Fang; Hu, Yaxi; Chen, Da; Li-Chan, Eunice C Y; Grant, Edward; Lu, Xiaonan

    2015-10-01

    Sudan I is a carcinogenic and mutagenic azo-compound that has been utilized as a common adulterant in spice and spice blends to impart a desirable red color to foods. A novel biosensor combining molecularly imprinted polymers (MIPs), thin layer chromatography (TLC) and surface enhanced Raman spectroscopy (SERS) could determine Sudan I levels in paprika powder to 1 ppm (or 2 ng/spot). Sudan I spiked paprika extracts (spiking levels: 0, 1, 5, 10, 40, 70 and 100 ppm) were prepared. Sudan I imprinted polymers were synthesized by employing the interaction between Sudan I (template) and methacrylic acid (functional monomer), followed by washing to remove Sudan I leaving the Sudan I-binding sites exposed. MIPs were used as a stationary phase for TLC and could selectively retain Sudan I at the original spot with little interference. A gold colloid SERS substrate could enhance Raman intensity for Sudan I in this MIP-TLC system. Principal component analysis plot and partial least squares regression (R(2)=0.978) models were constructed and a linear regression model (R(2)=0.983) correlated spiking levels (5, 10, 40, 70 and 100 ppm) with the peak intensities (721 cm(-1)) of Sudan I SERS spectra. Both separation (30-40s) and detection (1s or 0.1s) were extremely fast by using both commercial bench-top and custom made portable Raman spectrometers. This biosensor can be applied as a rapid, low-cost and reliable tool for screening Sudan I adulteration in foods.

  16. Determination of Sudan I in paprika powder by molecularly imprinted polymers-thin layer chromatography-surface enhanced Raman spectroscopic biosensor.

    PubMed

    Gao, Fang; Hu, Yaxi; Chen, Da; Li-Chan, Eunice C Y; Grant, Edward; Lu, Xiaonan

    2015-10-01

    Sudan I is a carcinogenic and mutagenic azo-compound that has been utilized as a common adulterant in spice and spice blends to impart a desirable red color to foods. A novel biosensor combining molecularly imprinted polymers (MIPs), thin layer chromatography (TLC) and surface enhanced Raman spectroscopy (SERS) could determine Sudan I levels in paprika powder to 1 ppm (or 2 ng/spot). Sudan I spiked paprika extracts (spiking levels: 0, 1, 5, 10, 40, 70 and 100 ppm) were prepared. Sudan I imprinted polymers were synthesized by employing the interaction between Sudan I (template) and methacrylic acid (functional monomer), followed by washing to remove Sudan I leaving the Sudan I-binding sites exposed. MIPs were used as a stationary phase for TLC and could selectively retain Sudan I at the original spot with little interference. A gold colloid SERS substrate could enhance Raman intensity for Sudan I in this MIP-TLC system. Principal component analysis plot and partial least squares regression (R(2)=0.978) models were constructed and a linear regression model (R(2)=0.983) correlated spiking levels (5, 10, 40, 70 and 100 ppm) with the peak intensities (721 cm(-1)) of Sudan I SERS spectra. Both separation (30-40s) and detection (1s or 0.1s) were extremely fast by using both commercial bench-top and custom made portable Raman spectrometers. This biosensor can be applied as a rapid, low-cost and reliable tool for screening Sudan I adulteration in foods. PMID:26078169

  17. Lipid Suppression in CSI with Spatial Priors and Highly Undersampled Peripheral k-space

    PubMed Central

    Bilgic, Berkin; Gagoski, Borjan; Kok, Trina; Adalsteinsson, Elfar

    2012-01-01

    Mapping 1H brain metabolites using chemical shift imaging (CSI) is hampered by the presence of subcutaneous lipid signals, which contaminate the metabolites by ringing due to limited spatial resolution. Even though CSI at spatial resolution high enough to mitigate the lipid artifacts is infeasible due to signal-to-noise (SNR) constraints on the metabolites, the lipid signals have orders of magnitude higher concentration, which enables the collection of high-resolution lipid maps with adequate SNR. The previously proposed dual-density approach exploits this high-SNR property of the lipid layer to suppress truncation artifacts using high-resolution lipid maps. Another recent approach for lipid suppression makes use of the fact that metabolite and lipid spectra are approximately orthogonal, and seeks sparse metabolite spectra when projected onto lipid-basis functions. The present work combines and extends the dual-density approach and the lipid-basis penalty, while estimating the high-resolution lipid image from 2-average k-space data to incur minimal increase on the scan time. Further, we exploit the spectral-spatial sparsity of the lipid ring and propose to estimate it from substantially undersampled (acceleration R = 10 in the peripheral k-space) 2-average in vivo data using compressed sensing, and still obtain improved lipid suppression relative to using dual-density or lipid-basis penalty alone. PMID:22807147

  18. Lipid suppression in CSI with spatial priors and highly undersampled peripheral k-space.

    PubMed

    Bilgic, Berkin; Gagoski, Borjan; Kok, Trina; Adalsteinsson, Elfar

    2013-06-01

    Mapping 1H brain metabolites using chemical shift imaging is hampered by the presence of subcutaneous lipid signals, which contaminate the metabolites by ringing due to limited spatial resolution. Even though chemical shift imaging at spatial resolution high enough to mitigate the lipid artifacts is infeasible due to signal-to-noise constraints on the metabolites, the lipid signals have orders of magnitude of higher concentration, which enables the collection of high-resolution lipid maps with adequate signal-to-noise. The previously proposed dual-density approach exploits this high signal-to-noise property of the lipid layer to suppress truncation artifacts using high-resolution lipid maps. Another recent approach for lipid suppression makes use of the fact that metabolite and lipid spectra are approximately orthogonal, and seeks sparse metabolite spectra when projected onto lipid-basis functions. This work combines and extends the dual-density approach and the lipid-basis penalty, while estimating the high-resolution lipid image from 2-average k-space data to incur minimal increase on the scan time. Further, we exploit the spectral-spatial sparsity of the lipid ring and propose to estimate it from substantially undersampled (acceleration R=10 in the peripheral k-space) 2-average in vivo data using compressed sensing and still obtain improved lipid suppression relative to using dual-density or lipid-basis penalty alone.

  19. Chemical Enhancer Solubility in Human Stratum Corneum Lipids and Enhancer Mechanism of Action on Stratum Corneum Lipid Domain

    PubMed Central

    Ibrahim, Sarah A.; Li, S. Kevin

    2010-01-01

    Previously, chemical enhancer-induced permeation enhancement on human stratum corneum (SC) lipoidal pathway at enhancer thermodynamic activities approaching unity in the absence of cosolvents (defined as Emax) was determined and hypothesized to be related to the enhancer solubilities in the SC lipid domain. The objectives of the present study were to (a) quantify enhancer uptake into SC lipid domain at saturation, (b) elucidate enhancer mechanism(s) of action, and (c) study the SC lipid phase behavior at Emax. It was concluded that direct quantification of enhancer uptake into SC lipid domain using intact SC was complicated. Therefore a liposomal model of extracted human SC lipids was used. In the liposome study, enhancer uptake into extracted human SC lipid liposomes (EHSCLL) was shown to correlate with Emax. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and differential scanning calorimetry (DSC) were used to evaluate lipid phase alterations in enhancer-treated intact SC. IR spectra demonstrated an increase in the lipid domain fluidity and DSC thermograms indicated a decrease in the phase transition temperature with increasing Emax. These results suggest that the enhancer mechanism of action is through enhancer intercalation into SC intercellular lipids and subsequent lipid lamellae fluidization related to enhancer lipid concentration. PMID:19747970

  20. Metabolism. Part III: Lipids.

    ERIC Educational Resources Information Center

    Bodner, George M.

    1986-01-01

    Describes the metabolic processes of complex lipids, including saponification, activation and transport, and the beta-oxidation spiral. Discusses fatty acid degradation in regard to biochemical energy and ketone bodies. (TW)