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Research article Special Issues

Ozone formation in relation with combustion processes in highly populated urban areas

  • The complex chain of photochemical reactions is one of the most important tasks in the air quality evaluation, expecially in urban areas. In fact, in this case there are high emission levels of NOx and no-methane hydrocarbons by combustion processes such as autovehicular traffic, domestic heating and industrial plants. Ozone is not emitted directly into the atmosphere but it is formed from a complex series of reactions between emitted nitrogen oxides (NOx) and reactive organic compounds (ROC). The high ozone concentrations, which occur during photochemical episodes, are usually accompanied by elevated concentrations of other photochemical oxidants such as nitric acid (HNO3), peroxyacylnitrates (PANs), hydrogen peroxide (H2O2), etc. The complex series of these reactions constitutes the most important issue to the degradation of air quality. Further, the NMHCs play a key role in the formation of photochemical air pollution: they are considered as precursors for ozone production at the ground level when the sunlight and nitrogen oxides are present. From a practically point of view defining a quality standard or a limit is substantially correct but it is no sufficient to solve the problem. So it should become necessary to acquire knowledge on the different formation mechanisms of the photochemical pollution phenomena. In this paper there will be shown the results of a long-term study performed in Rome for evaluating the ozone formation in relationship with the autovehicular traffic density.

    Citation: Pasquale Avino, Maurizio Manigrasso. Ozone formation in relation with combustion processes in highly populated urban areas[J]. AIMS Environmental Science, 2015, 2(3): 764-781. doi: 10.3934/environsci.2015.3.764

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  • The complex chain of photochemical reactions is one of the most important tasks in the air quality evaluation, expecially in urban areas. In fact, in this case there are high emission levels of NOx and no-methane hydrocarbons by combustion processes such as autovehicular traffic, domestic heating and industrial plants. Ozone is not emitted directly into the atmosphere but it is formed from a complex series of reactions between emitted nitrogen oxides (NOx) and reactive organic compounds (ROC). The high ozone concentrations, which occur during photochemical episodes, are usually accompanied by elevated concentrations of other photochemical oxidants such as nitric acid (HNO3), peroxyacylnitrates (PANs), hydrogen peroxide (H2O2), etc. The complex series of these reactions constitutes the most important issue to the degradation of air quality. Further, the NMHCs play a key role in the formation of photochemical air pollution: they are considered as precursors for ozone production at the ground level when the sunlight and nitrogen oxides are present. From a practically point of view defining a quality standard or a limit is substantially correct but it is no sufficient to solve the problem. So it should become necessary to acquire knowledge on the different formation mechanisms of the photochemical pollution phenomena. In this paper there will be shown the results of a long-term study performed in Rome for evaluating the ozone formation in relationship with the autovehicular traffic density.


    1. Introduction

    The plasma membrane is the main permeability barrier of the cell and consists of hundreds to thousands of different lipid species in addition to a wide range of proteins that allow the cell to sense the environment and transport specific molecules in and out of the cell. The lipids of the membrane are not randomly distributed but can form distinct domains, often referred to as lipid rafts, and associate with specific proteins [1,2,3]. Rafts are associated with specific membrane proteins, thereby affecting signaling and protein trafficking in the membrane as summarized by Levental and Veatch [4].

    Hydrocarbons affect the membrane properties as they interfere with the interaction of proteins with their neighboring lipids. Alternatively, the hydrocarbons can bind to hydrophobic pockets or surfaces of proteins and thereby influence their activity. Local anesthetics exert their effects by e.g. decreasing the miscibility temperature of lipids as shown in giant plasma membrane vesicles [5], thereby increasing the membrane fluidity. In another study, hydrophobic phytochemicals were shown to perturb the phospholipid bilayer and the proteins embedded in there [6]. In general, hydrocarbons alter membrane properties such as membrane thickness, head group hydration and fluidity, all of which can affect membrane proteins [7].

    The toxicity of hydrocarbons and other molecules is frequently related to the hydrophobicity of the compounds. A measure for hydrophobicity is the logP value, the partitioning of a molecule over octanol and water. The more hydrophobic the compound (as indicated by a higher, positive logP value), the more it partitions in octanol and accordingly the higher the concentration in the membrane [8,9]. For instance, 20 mg of petroleum hydrocarbons per gram lipids have been found in oysters [10] and 93 μg/g lipid in maple leaves [11]. Organisms respond to hydrophobic pollutants by changing their membrane composition, by degrading PAHs and by expressing efflux pumps to expel the molecules from the membrane [7,12,13]. It has been shown that Escherichia coli and Ralstonia eutropha cells change their lipid saturation to make up for the fluidizing or ordering effects of the pollutant when exposed to phenol or biphenyl [14,15].

    Aliphatic hydrocarbons localize in the central part of the bilayer [16,17]. Molecular dynamics (MD) simulations confirm experimental studies and found that aliphatic hexane [18] and ethane [19] reside in the hydrophobic center of the bilayer. As a general rule, amphipathic molecules partition near the bilayer interface, while more hydrophobic molecules reside near the bilayer center. In the center of the bilayer, aliphatic hydrocarbons interact with the acyl chains of the phospholipids and increase the area occupied by a phospholipid [20]. This localization prevents Van der Waals interactions between neighboring lipids, thereby fluidizing the membrane. In contrast, long chain alkanes interdigitate between the leaflets, thereby increasing the overall degree of ordering in the membrane [16].

    The effects of cyclic hydrocarbons on biological membranes were studied extensively in the early 90's [8], reviewed in Sikkema et al., 1995. It was found that the partitioning in the membrane of cyclic hydrocarbons scales linearly with the logP values of the molecules and they expand the membrane [7]. In membrane vesicles derived from Escherichia coli cells the hydrocarbons thicken the bilayer and increased the membrane fluidity. In addition, the membranes became more permeable to protons, and, accordingly, it became more difficult to maintain a proton motive force. It was then concluded that global deformation of the membrane likely accounts for the toxicity effects.

    Polycyclic aromatic hydrocarbons (PAHs) are found as pollutants in the environment, mainly as a result of incomplete combustion. PAHs are very stable and persistent once formed, and they may accumulate in the center of lipid bilayers [7]. Such localization was found for the aromatic benzene [19,21,22] and pyrene [23,24,25]. Simulation data on the interaction of small, aromatic molecules are described in [26]. The toxicity of PAHs in eukaryotes is dual and relates in part to their hydrophobicity. First, these molecules accumulate in lipid membranes and affect membrane function. Second, to remove these compounds from the cell membrane, the PAHs are chemically activated by epoxidation, but the modified compounds can also react with other molecules in the cell such as DNA. Depending on where the PAH epoxidation takes place, these metabolites are carcinogenic [27].

    Biological membranes are heterogeneous and consist of domains [28] that are on the nanometer scale and short-lived [29], making it a challenge to study their properties. We use giant unilamellar vesicles (GUVs) with detergent-resistant membrane domains (DRMs) as model systems to study mixing effects of hydrocarbons on lipid domain formation. Phase-separating GUVs can be made from a minimum of three components: typically a saturated lipid, an unsaturated lipid and a sterol. At the right ratio of lipids, the GUVs have a liquid-ordered (Lo) phase, enriched in the saturated lipid and cholesterol, and a liquid-disordered (Ld) phase, mainly consisting of the unsaturated lipid [30,31]. Detergent-resistant membranes (DRMs) derived from phase-separating vesicles are closely related to the Lo domains. Lipids associated with the Lo phase were enriched in DRMs [32], and the DRM fraction can only be obtained from vesicles that are phase-separating or in the Lo phase [33,34]. In addition, the Lo phase of phase-separating supported bilayers was found detergent resistant [35]. These model membranes mimic the behavior of natural lipid mixtures [36,37,38,39].

    In previous work, the aromatic Lo preferring dye naphtopyrene was found to perturb the membrane around the miscibility transition temperature at concentrations of 0.3 mol% [40]. A recent molecular dynamics study by Barnoud and coworkers [41] indicated a difference between the effects of aromatic and aliphatic compounds. While aliphatic compounds induced mixing of a phase-separating membrane, aromatic hydrocarbons stabilized the phase separation.

    To better understand the toxicity of PAHs in eukaryotic cells, we determined their effects (Figure 1) on the lipid phase separation in GUVs. We benchmarked the effects of aromatic compounds of varying size against unsubstituted aliphatic compounds as the molecules are expected to interact differently with lipids and are expected to partition in different places of the lipid bilayer. Indeed, we find that the effects on phase separation are highly dependent on the partitioning behavior of the hydrocarbons. Furthermore, we find differences for membranes with DPPC or SSM as the saturated lipid component, indicating that subtle variations in the membrane lipid composition can have major impact when membrane-active compounds are present in the environment. The lipid mixing effect of PAHs and differences between experiments and simulations are discussed and put in perspective.

    Figure 1. Structures of the compounds used in this study.

    2. Materials and Methods


    2.1. Materials

    DPPC, SSM, DOPC and cholesterol were purchased from Avanti Polar Lipids. ATTO 550 DOPE and ATTO 655 DOPE were used as fluorescent probes to visualize the Ld phase and obtained from ATTO-Tec. The dyes are both hydrophilic but differ in their charge (cationic versus zwitterionic). We used both dyes to minimize the possibility of artifacts due to interactions between dye and lipids or and dye and PAHs. The hydrocarbons naphthalene, tetracene, chrysene, pyrene, perylene, triphenylene, coronene, octane and hexadecane were purchased from Sigma-Aldrich, and of fluorescence grade when available. Corannulene was purchased from TCI Europe. Structures of the hydrocarbons used in this study are presented in Figure 1 and their properties are listed in Table 1.

    Table 1. Properties of hydrocarbons used in this study.
    Compound Molecular formula Mw (g/mol) Boiling point (℃) log Pc XLog P3d Absorption max (nm) Absorption max (nm)a Em max (nm)a
    Naphthalene C10H8 128.17052 218 3.3/3.35 3.3 221,275.5,286,311 220,275,286,311 322,334
    Phenanthrene C14H10 178.2292 340 4.46 4.5 210,219,242,251,273.5,281,292.5,308.5,314,322.5,329.5,337,345
    Tetracene C18H12 228.28788 450b 5.76–6.02b 5.9
    Chrysene C18H12 228.28788 448 5.73/5.9 5.7 222,258,268,295,353,361,344,320
    Pyrene C16H10 202.2506 404/399 4.88 4.9 273,306,320,335 241,273,335 349,381
    Triphenylene C18H12 228.28788 425b 4.83–5.84b 4.9
    Benzo(e)pyrene C20H12 252.30928 310–312 6.44 6.4
    Perylene C20H12 252.30928 350–400 (sublimes) 5.82 5.8 245,251,368,387,406,434 387,408,436 436,463,497
    Corannulene C20H10 250.2934 6
    Coronene C24H12 300.35208 525b 5.4–8.2b 7.2
    Octane C8H18 114.22852 126 5.18 3.9
    Hexadecane C16H34 226.44116 286.5 8.25 (est) 8.3
    Data from Pubchem database, except ahttp://omlc.org/spectra/PhotochemCAD/index.html; bMackay D, Shiu WY, Ma KC, et al. (2006) Handbook of Physical-Chemical Properties and Environmental Fate for Organic Chemicals, 2 Eds., CRC Press. clogP = log ([solute]octanol/[solute]water); dXlogP3 = a calculated logP value [57].
     | Show Table
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    2.2. GUV formation

    GUVs were prepared by electroformation as described previously [31]. Lipid mixtures consisting of DPPC/DOPC/cholesterol or SSM/DOPC/cholesterol in a 4:3:3 ratio (all in chloroform/methanol 9:1) were prepared out of 5 mM stocks. To visualize the GUVs, 0.1% ATTO 550 DOPE or ATTO 655 DOPE was added. 15 μL of the lipid mixture was placed on a conductive indium tin oxide (ITO) coated glass plate. Solvents were removed by placing the coverslips with lipids in a vacuum desiccator for 1 h. A rubber ring (Ø15 mm) was placed around the lipids with grease. After preheating the glass plates and water to 50 ℃, the ITO-plate containing the lipids was placed on the Vesicle Prep Pro (Nanion Technologies). 200 mL water was added and the chamber was closed by putting a second ITO plate on top. A voltage of 1.1 V was applied for 1 h, at 10 Hz and 50 ℃ to form the GUVs. Afterwards, the chamber was disassembled and the GUVs were studied by confocal microscopy.


    2.3. Addition of hydrocarbons

    Hydrocarbons dissolved in chloroform/methanol 9:1 or when indicated in dimethylformamide (DMF) were added to the lipid mixture or to GUVs. As solvent control, the maximal solvent concentration was taken as extra condition. To study the effect of hydrocarbons, the compounds dissolved in chloroform/methanol 1:1 were added to the lipid mixture. The GUVs formed were imaged on a commercial LSM 710 confocal microscope (Zeiss), using a 40× C-Apochromat Corr M27 with NA 1.2 water immersion objective. ATTO 550 DOPE was excited with a 543 nm HeNe laser, ATTO 655 DOPE with a 633 nm HeNe laser. Perylene was excited with a 405 nm diode laser.


    2.4. Data analysis

    To quantify the effect of hydrocarbons on phase separation, the partitioning of the dyes over the Lo and Ld phases was used and reported as pLo/Ld ratio. This ratio is equivalent to the partitioning coefficient (Kp) that was used by Levental and coworkers [1]. A 5 pixel wide line was drawn through the middle of a GUV to avoid polarization effects, as shown in Figure 2. The maxima of both peaks were determined and the pLo/Ld was calculated. At least 50 GUVs per condition for each experiment were analysed.


    2.5. Detergent-resistant membranes (DRMs)

    To probe the partitioning of the PAHs, DRMs were prepared from multilamellar vesicles as previously described [42], with slight modifications. Briefly, multilamellar vesicles were formed by thin film hydration. The appropriate amount of lipids, dissolved in chloroform/methanol 9:1, were mixed and solvents were evaporated by rotary evaporation. Next, the lipid film was hydrated in 10 mM Tris-HCl, 150 mM NaCl, pH 7.4 by repeated vortexing at 60 ℃; the final lipid concentration was made 1 mM. To isolate DRMs, ice cold Triton X-100 was added to chilled MLVs in a 1:1 mol Triton X-100 to lipid ratio. These conditions were chosen to observe similar perylene partitioning in the vesicles with DRMs as in the GUVs with Ld and Lo phases (see Figure 5). After 30 minutes of incubation on ice, the DRMs were obtained by ultracentrifugation at 227,000 g for 1 h at 4 ℃. The supernatant was removed and the pellet resuspended in the same volume of Tris/NaCl buffer. Fluorescence of the pellet and the supernatant was measured on a fluorimeter (Jasco FP-8300).

    Figure 5. PAH localization in detergent resistant membranes. The Ipellet/Isupernatant was calculated from the fluorescence of the pellet (DRM) and the fluorescence of the supernatant at the maximum emission. All compounds were present at 2 mol% hydrocarbon-to-lipid ratio to prevent excimer formation. Values are mean ± standard deviation of at least two independent experiments.

    3. Results


    3.1. Pyrene and related compounds prevent phase separation

    Pyrene, triphenylene and benzo(e)pyrene prevented phase separation in GUV, composed of DPPC, DOPC and cholesterol when added to the lipid mixture in a 1 to 1 molar ratio (Figure 3A). The other tested aromatic hydrocarbons, i.e. naphthalene, phenanthrene, tetracene, chrysene, perylene, coronene and corannulene, retained phase separation, even at such high concentrations. Also for the aliphatic octane and hexadecane, no effect on phase separation was observed. The majority of the GUVs are either phase separating (indicated by a pLo/Ld close to 0) or uniform (indicated by a pLo/Ld close to 1). In the GUVs analyzed, few vesicles displayed an intermediate appearance between phase separation and one phase (where phase separation is maintained, but the dye partitioning is not as black and white as in the example shown in Figure 2), which is indicated by a pLo/Ld value between 0.2 and 0.8 (Figure 3B).

    Figure 2. Fluorescent quantification of pLo/Ld. Partition coefficients of the dyes were quantified by a 5 pixel width line scan through the domains. Only GUVs with both domains in the middle (as in the left picture) were analyzed. When no phase separation was visible, a line was drawn from left to right through the middle of the GUV (as in the right panel).
    Figure 3. Pyrene and related molecules prevent phase separation in GUVs composed of DPPC/DOPC/cholesterol. A: GUVs composed of DPPC/DOPC/cholesterol at a ratio of 4:3:3 and the solutes dissolved in chloroform/methanol were used. The pLo/Ld ratio was determined using ATTO 550 DOPE as probe and the hydrocarbons were added to the lipid mixture prior to GUV formation. The error reflects variations in different GUV preparations. All compounds are present in a 1 to 1 mol ratio with the lipids. In green: aromatic hydrocarbons; in red: aliphatic hydrocarbons. B: Distribution plot of one representative experiment, for three conditions. pLo/Ld values of individual GUVs (indicated by a symbol) are ordered from 0 (lowest pLo/Ld ratio measured for that condition) to 1 (highest pLo/Ld ratio measured) according to the their pLo/Ld ratio; the pLo/Ld ratios are plotted against the GUV number. We normalized the values of the x-axis, because the GUV numbers are not the same for the three conditions. Black line: 2.5 mol% pyrene; red line: 10 mol% pyrene; green line: 50 mol% pyrene. In A, values are mean ± standard deviation of at least three independent experiments (biological replicates) except for naphthalene, tetracene, coronene, octane and hexadecane (n = 2), and triphenylene and corannulene (n = 4).

    Irrespective of whether the hydrocarbon was introduced prior to or after GUV formation, pyrene dissipated phase separation in the GUVs (Figure S1A). Adding pyrene dissolved in DMF to phase-separating GUVs increased the pLo/Ld from 0.07 to 0.86. Various fluorescent probes, used to visualize membranes, have been shown to alter the miscibility temperature of membranes [40,43,44,45]. Therefore, to rule out possible effects of the cationic membrane probe (ATTO 550 DOPE), the experiments were repeated with the zwitterionic ATTO 655 DOPE but the results were similar (see Figure S1B).


    3.2. Phase separation only disappears at high PAH to lipid ratios and is lipid composition dependent

    To study if the mixing effect of pyrene is lipid specific, the effect of pyrene was also studied in GUVs prepared from SSM/DOPC/cholesterol (Figure 4). At similar pyrene to lipid ratios, phase separation was maintained in SSM/DOPC/cholesterol GUVs but not in vesicles prepared from DPPC/DOPC/cholesterol. These results are consistent with previous measurements [46,47,48,49], which showed that the interaction between SSM and cholesterol is stronger than the interaction between DPPC and cholesterol. Accordingly, the impact of pyrene and most likely other PAHs on phase separation is clear when DPPC is present, in contrast with the sphingolipid.

    Figure 4. Phase separation disappears at high PAH to lipid ratios and is dependent on lipid composition. The pLo/Ld ratio estimated from the ATTO 550 DPPE partitioning in GUVs composed of DPPC/DOPC/cholesterol or SSM/DOPC/cholesterol (mol ratios of 4:3:3) with and without the indicated mol% of pyrene. Values are mean ± standard deviation of at least two independent experiments.

    3.3. PAH localization depends on hydrophobicity and shape

    The localization of PAHs was studied in DRMs, since these resemble the Lo phase and PAH partitioning can be determined spectroscopically. Here, we observe that the more hydrophobic the compound (as indicated by the logP values; Table 1) the higher the partitioning in the DRM (Figure 5). Small PAHs such as naphthalene and phenanthrene have a preference for the Ld phase (indicated by the Ipellet/Isupernatant < 1), while the larger compounds tetracene and coronene reside mainly in the Lo phase (Ipellet/Isupernatant > 1). Strikingly, with the exception of corannulene, the three compounds that prevent phase separation in GUVs equally partitioned in both phases (Ipellet/Isupernatant ≈ 1). To check if the partitioning of hydrocarbons in DRMs is comparable to partitioning in GUVs, the localization of perylene was tested by an independent method. Perylene absorbs blue light and has a fluorescence emission maximum at 436 nm and can therefore be followed by confocal microscopy. The fluorescence-based analyses in GUVs were compared to the results from DRMs (Figure 6), and indeed a similar localization was found.

    Figure 6. Perylene localization in GUVs and DRMs. A: the pLo/Ld ratio of perylene in both DPPC/DOPC/cholesterol and SSM/DOPC/cholesterol GUVs (mol ratios of 4:3:3) GUVs; 2 mol% perylene was added to the lipid mixture prior to GUV formation. B: the Ipellet/Isupernatant of perylene were determined in multilamellar vesicles of the aforementioned lipid mixtures with 2 mol% perylene. Values are mean ± standard deviation of at least two independent experiments.

    4. Conclusions

    We find that at room temperature high concentrations of the hydrocarbons naphthalene, phenanthrene, tetracene, chrysene, perylene, corannulene, corulene, octane and hexadecane have a rather small effect on lipid phase separation in vesicles composed of DPPC, DOPC and cholesterol. Differences in phase separation are not visible even when hydrocarbons are present in amounts stoichiometric with the membrane lipids. Pyrene, benzo(e)pyrene and triphenylene form an exception, in that these compounds induce lipid mixing in phase-separating GUVs containing DPPC but not when DPPC is replaced by SSM. The specific effect of pyrene-like compounds is likely due to their partitioning in both the Lo and Ld phase, which is explained by the shape and hydrophobicity of the hydrocarbon.

    According to MD simulations and fluorescence quenching experiments, pyrene is localized predominantly in the highly ordered upper region of the acyl chains of POPC/DPPC membranes [23,24], at a similar position as cholesterol [50]. Pyrene does not reach as deep as cholesterol into the bilayer, thereby leaving space below the pyrene molecule and the center of the membrane. The tails of unsaturated lipids such as DOPC can occupy this space [50]. Hexadecane is located in a similar fashion as pyrene according to X-ray diffraction data [16]. On the contrary, octane is localized between the two leaflets in the same study. To the best of our knowledge, for the other compounds used in this study no localization data is available.

    Besides its position in the upper region of the acyl chains, pyrene has more in common with cholesterol. In MD simulations, pyrene had an ordering effect on neighboring DPPC molecules in the fluid phase, while it has a disordering effect on the same molecules in the gel phase [21]. This is similar to the effect of cholesterol in DPPC membranes [51]. In addition, pyrene has a diamond shape and occupies the equivalent geometric volume of the membrane [50]. Compared to e.g. tetracene or chrysene, more space is available below the pyrene molecule. If indeed pyrene behaves as cholesterol, the membrane becomes saturated and differences between the Lo and the Ld phase become smaller. Eventually, both phases mix as seen in ternary lipid mixtures (e.g. DPPC, DOPC, and cholesterol [30]) that contain over 40% cholesterol and this is what we find here with pyrene.

    Large PAHs have a preference for the Lo phase [39], while benzene and fullerene end up in the Ld phase of phase-separating bilayers in MD simulations [41]. This is in agreement with the localization of PAHs in DRMs measured here. The large rigid compounds induce order by forcing the acyl chains to arrange themselves around the molecule, which occurs with an entropic penalty [24]. In the already more ordered Lo phase, the costs are lower than in less ordered Ld phase, hence the preference of these compounds for Lo.

    The exact localization of pyrene in phase-separating membranes has not been reported but can be deduced from literature using similar compounds. The partitioning of aromatic dyes is not only dependent of their hydrophobicity but also of their size and shape. Relatively small dyes such as perylene and rubicene were found in both the Lo and Ld phase of GUVs composed of brain SM, DOPC and cholesterol, larger dyes such as terrylene and naphthopyrene partitioned in the Lo phase [52]. Naphthopyrene also partitions into the Lo phase of GUVs composed of DPPC/DOPC/cholesterol [53]. However, the dye phase preference varies between lipid mixtures. For example perylene has Lo preference in GUVs composed of egg SM (mainly consisting of short chain (C16) saturated SM), DOPC and cholesterol [54], while in brain SM (consisting of longer chain SMs (C18 to C24) and 20% unsaturated chains), DOPC and cholesterol GUVs perylene does not have a preference for any of the phases [52]. These studies indicate that the more hydrophobic the dye, the more likely it is that it localizes in the Lo phase, but only few very hydrophobic compounds end up in that Lo phase and it depends on the lipid mixture used how a dye is distributed across both phases (a Lo phase in one lipid mixture is different from a Lo phase in another lipid mixture).

    The dissipation of phase separation with pyrene and related compounds was only observed in vesicles containing DPPC. Although it is often claimed that DPPC and SM act similar in phase separating mixtures, the strength of the interaction of these lipids with e.g. cholesterol is different. The preference of cholesterol for SM is explained by the presence of the N-linked acyl chain. The amide of SM can act as hydrogen bond donor and acceptor with the hydroxyl moiety of the cholesterol [55]. Due to the stronger interactions between cholesterol and SSM, pyrene most likely cannot perturb phase separation, i.e. under conditions that it does in GUVs with DPPC instead of SSM. Other studies have shown different partitioning of the dye DiI C18:0, depending on the saturated lipid component. The DiI C18:0 dye partitions into the Ld phase of brain SM-containing GUVs and in the Lo phase in distearoylphosphatidylcholine-containing GUVs [52]. The authors explain this effect due to the preferential interaction of cholesterol with SM (excluding the DiI C18:0 from this phase) compared to saturated phospholipids. This is also confirmed by 2H-NMR [46,49], solid-state NMR combined with DSC [48] and DPH anisotropy measurements, using a fluorescent cholesterol analogue [47].

    Aliphatic hydrocarbons had no effect on phase separation in GUVs composed of DPPC, DOPC and cholesterol, analyzed at room temperature. This is in contrast to previous MD simulations, where these molecules act as lineactant and decrease phase separation [41]. We attribute the differences in the experiments and simulations to either differences in lipid composition (the simulation studies use polyunsaturated lipids to increase the phase separation) or setup (small periodic lamellar patches with a surface in the order of 520 nm2 in case of the MD simulations versus GUVs in the experiments). An older study found that the aromatic benzene and toluene increase membrane fluidity, but the aliphatic cyclohexane and hexane did not alter membrane fluidity as measured by pyrene excimer formation [56]. This is in line with the results presented here, where only some aromatic compounds alter phase separation.

    In conclusion, we show that at room temperature hydrocarbons have a distinct effect on lipid phase separation, and the effect is dependent on the strength of the interaction of cholesterol with the saturated lipid component. Pyrene and pyrene-like compounds dissipate phase separation in mixtures containing DPPC as saturated lipid component but not in GUVs containing SSM instead of DPPC. We speculate that pyrene and related compounds act as cholesterol, thereby decreasing the difference between the Lo and Ld phase and eventually leading to domain mixing. Furthermore, PAHs larger than pyrene-like compounds prefer Lo, whereas smaller ones partition in Ld.


    Acknowledgements

    This work was supported by the Netherlands Organisation for Scientific Research (NWO): Chem-Them grant 728.011.202. Prof. Gerard Roelfes and Hugo van Oosterhout are kindly acknowledged for fruitful discussions.


    Conflict of Interest

    All authors declare no conflicts of interest in this paper.


    [1] Filanyson-Pitts BJ, Pitts JN Jr (1986) Atmospheric Chemistry Fundamentals and Experimental Techniques, New York: Wiley.
    [2] Olaguer EP, Rappenglück B, Lefer B, et al. (2009) Deciphering the role of radical sources during the Second Texas Air Quality Study. J Air Waste Man Assoc 59: 1258-1277. doi: 10.3155/1047-3289.59.11.1258
    [3] Rappenglück B, Oyola P, Olaeta I, et al. (2000) The evolution of photochemical smog in the Metropolitan Area of Santiago de Chile. J Appl Meteor 39: 275-290. doi: 10.1175/1520-0450(2000)039<0275:TEOPSI>2.0.CO;2
    [4] Rappenglück B, Schmitza R, Bauerfeind M, et al. (2005) An urban photochemistry study in Santiago de Chile. Atmos Environ 39: 2913-2931. doi: 10.1016/j.atmosenv.2004.12.049
    [5] Czader BH, Byuna DW, Kima S-T (2008) A study of VOC reactivity in the Houston-Galveston air mixture utilizing an extended version of SAPRC-99 chemical mechanism. Atmos Environ 42: 5733-5742. doi: 10.1016/j.atmosenv.2008.01.039
    [6] Paulson SE, Orlando JJ (1996) The reactions of ozone with alkenes: an important source of HOx in the boundary layer. Geophys Res Lett 23: 3727-3730. doi: 10.1029/96GL03477
    [7] Wolff S, Boddenberg A, Thamm J, et al. (1997) Gas-phase ozonolysis of ethene in the presence of carbonyl-oxide scavengers. Atmos Environ 31: 2965-2969. doi: 10.1016/S1352-2310(97)00114-3
    [8] Osborn DL, Hyeon C, Mordaunt DH, et al. (1997) Fast beam photodissociation spectroscopy and dynamics of the vinoxy radical. J Chem Phys 106: 3049-3066. doi: 10.1063/1.473419
    [9] Calvert JG, Atkinson R, Kerr JA, et al. (2000) The Mechanism of atmospheric oxidation of the alkenes. Oxford University Press, New York.
    [10] Ryerson TB, Trainer M, Angevine WM, et al. (2003) Effect of petrochemical industrial emissions of reactive alkenes and NOx on tropospheric ozone formation in Houston, Texas. J Geophys Res 108: 4249-4273. doi: 10.1029/2002JD003070
    [11] Chan WT, Hamilton IP (2003) Mechanisms for the ozonolysis of ethene and propene: Reliability of quantum chemical predictions. J Chem Phys 118: 1688-1701. doi: 10.1063/1.1531104
    [12] Topaloglou C, Kazadzis S, Bais AF, et al. (2005) NO2 and HCHO photolysis frequencies from irradiance measurements in Thessaloniki, Greece. Atmos Chem Phys 5: 1645-1653. doi: 10.5194/acp-5-1645-2005
    [13] Johnson D, Marston G (2008) The gas-phase ozonolysis of unsaturated volatile organic compounds in the troposphere. Chem Soc Rev 37: 699-716. doi: 10.1039/b704260b
    [14] Volkamer R, Sheehy P, Molina LT, et al. (2010) Oxidative capacity of the Mexico City atmosphere - Part 1: A radical source perspective. Atmos Chem Phys 10: 969-6991.
    [15] Su H, Cheng Y, Oswald R, et al. (2011) Soil nitrite as a source of atmospheric HONO and OH radicals. Science 333: 1616-1618. doi: 10.1126/science.1207687
    [16] Alam MS (2011) Total radical production and degradation products from alkene ozonolysis. Ph.D. Thesis (available at http: //etheses.bham.ac.uk/1733/2/Alam_11_PhD.pdf; accessed on May 2015).
    [17] Alam MS, Camredon M, Rickard AR, et al. (2011) Total radical yields from tropospheric ethene ozonolysis. Phys Chem Chem Phys 13: 11002-11015. doi: 10.1039/c0cp02342f
    [18] Edwards PM, Young CJ, Aikin K, et al. (2013) Ozone photochemistry in an oil and natural gas extraction region during winter: simulations of a snow-free season in the Uintah Basin, Utah. Atmos Chem Phys 13: 8955-8971. doi: 10.5194/acp-13-8955-2013
    [19] Vrekoussis M, Mihalopoulos N, Gerasopoulos N, et al. (2007) Two-years of NO3 radical observations in the boundary layer over the Eastern Mediterranean. Atmos Chem Phys 7: 315-327. doi: 10.5194/acp-7-315-2007
    [20] Rollins AW, Kiendler-Scharr A, Fry J, et al. (2009) Isoprene oxidation by nitrate radical: alkyl nitrate and secondary organic aerosol yields. Atmos Chem Phys 9: 6685-6703. doi: 10.5194/acp-9-6685-2009
    [21] Jorquera H, Rappenglück B (2004) Receptor modeling of ambient VOC at Santiago, Chile. Atmos Environ 38: 4243-4263. doi: 10.1016/j.atmosenv.2004.04.030
    [22] Elshorbany YF, Kurtenbach R, Wiesen P, et al. (2009) Oxidation capacity of the city air of Santiago, Chile. Atmos Chem Phys 9: 2257-2273. doi: 10.5194/acp-9-2257-2009
    [23] Elshorbany YF, Kleffmann J, Kurtenbach R, et al. (2009) Summertime photochemical ozone formation in Santiago, Chile. Atmos Environ 43: 6398-6407. doi: 10.1016/j.atmosenv.2009.08.047
    [24] Czader BH, Li X, Rappenglück B (2013) CMAQ modeling and analysis of radicals, radical precursors, and chemical transformations. J Geophys Res 118: 11376-11387.
    [25] Czader BH, Rappenglück B (2015) Modeling of 1,3-butadiene in urban and industrial areas. Atmos Environ 102: 30-42. doi: 10.1016/j.atmosenv.2014.11.039
    [26] Riedel TP, Wolfe GM, Danas KT, et al. (2014) An MCM modeling study of nitryl chloride (ClNO2) impacts on oxidation, ozone production and nitrogen oxide partitioning in polluted continental flow. Atmos Chem Phys 14: 3789-3800. doi: 10.5194/acp-14-3789-2014
    [27] Mao J, Rena X, Chen S, et al. (2010) Atmospheric oxidation capacity in the summer of Houston 2006: comparison with summer measurements in other metropolitan studies. Atmos Environ 44: 4107-4115. doi: 10.1016/j.atmosenv.2009.01.013
    [28] Elshorbany YF, Kleffmann J, Kurtenbach R, et al. (2010) Seasonal dependence of the oxidation capacity of the city of Santiago de Chile. Atmos Environ 44: 5383-5394. doi: 10.1016/j.atmosenv.2009.08.036
    [29] European Commission, Directive 2008/50/EC of the European Parliament and of the Council of 21 May 2008 on ambient air quality and cleaner air for Europe. 2008. Available from: http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=celex:32008L0050#text
    [30] Mustafa MG (1990) Biochemical basis of ozone toxicity. Free Radical Biol Med 9: 245-265. doi: 10.1016/0891-5849(90)90035-H
    [31] Hazuchaa MJ, Lefohn AS (2007) Nonlinearity in human health response to ozone: experimental laboratory considerations. Atmos Environ 41: 4559-4570. doi: 10.1016/j.atmosenv.2007.03.052
    [32] Pryor WA, Squadrito GL, Friedman M (1995) The cascade mechanism to explain ozone toxicity: the role of lipid ozonation products. Free Radical Biol Med 19: 935-941. doi: 10.1016/0891-5849(95)02033-7
    [33] Stockinger HE (1965) Ozone toxicology: a review of research and industrial experience. Arch Environ Health 10: 719-731. doi: 10.1080/00039896.1965.10664082
    [34] Avino P, De Lisio V, Grassi M, et al. (2004) Influence of air pollution on Chronic Obstructive Respiratory Diseases: comparison between city (Rome) and hillcountry environments and climates. Ann Chim (Rome) 94: 629-636. doi: 10.1002/adic.200490080
    [35] Ferretti M, Fagnano M, Amoriello T, et al. (2007) Measuring, modelling and testing ozone exposure, flux and effects on vegetation in southern European conditions - What does not work? A review from Italy. Environ Poll 146: 648-658.
    [36] Platt U, Perner D, Patz HW (1979) Simultaneous measurement of atmospheric CH2O, O3 and NO2 by differential optical absorption. J Geophys Res 84: 6329-6335. doi: 10.1029/JC084iC10p06329
    [37] Morales JA, Treacy J, Coffey S (2004) Urban ozone measurements using differential optical absorption spectroscopy. Anal Bioanal Chem 379, 51-55.
    [38] Avino P, Manigrasso M (2008) Ten-year measurements of gaseous pollutants in urban air by an open-path analyzer. Atmos Environ 42: 4138-4148. doi: 10.1016/j.atmosenv.2008.01.024
    [39] Avino P, Brocco D, Lepore L (2001) Determination of atmospheric organic and elemental carbon particle in Rome with a thermal method. Anal Lett 34: 967-974. doi: 10.1081/AL-100103606
    [40] Avino P, Brocco D, Cecinato A, et al. (2002) Carbonaceous component in atmospheric aerosol: measurement procedures and characterization. Ann Chim (Rome) 92: 333-341.
    [41] Avino P, Brocco D, Lepore L, et al. (2000) Distribution of elemental carbon (EC) and organic carbon (OC) in the atmospheric aerosol particles of Rome. J Aerosol Sci 31: S364-S365. doi: 10.1016/S0021-8502(00)90375-6
    [42] Avino P, Brocco D, Lepore L, et al. (2003) Interpretation of atmospheric pollution phenomena in relationship with the vertical atmospheric remixing by means of natural radioactivity measurements (Radon) of particulate matter. Ann Chim (Rome) 93: 589-594.
    [43] Febo A, Guglielmi F, Manigrasso M, et al. (2010) Local air pollution and long-range mass transport of atmospheric particulate matter: a comparative study of the temporal evolution of the aerosol size fractions. Atmos Poll Res 1: 141-146. doi: 10.5094/APR.2010.018
    [44] Manigrasso M, Febo A, Guglielmi F, et al. (2012) Relevance of aerosol size spectrum analysis as support to qualitative source apportionment studies. Environ Poll 170: 43-51. doi: 10.1016/j.envpol.2012.06.002
    [45] Avino P, Manigrasso M, Cuomo F (2015) Natural radioactivity as an easy and quick parameter for describing the dynamic of the Planetary Boundary Layer. RSC Adv 5: 57538-57549. doi: 10.1039/C5RA10618D
    [46] Law n. 413 of 04/11/1997. Misure urgenti per la prevenzione dell'inquinamento atmosferico da benzene. Gazzetta Ufficiale Italiana n. 282 of 03/12/1997.
    [47] Rappenglück B, Lubertino G, Alvarez S, et al. (2013) Radical precursors and related species from traffic as observed and modeled at an urban highway junction. J Air Waste Man Assoc 63: 1270-1286. doi: 10.1080/10962247.2013.822438
    [48] Monod A, Sive BC, Avino P (2001) Monoaromatic compounds in ambient air of various cities: a focus on correlations between the Xylenes and Ethylbenzene. Atmos Environ 35: 135-149. doi: 10.1016/S1352-2310(00)00274-0
    [49] De Lauretis R, Ilacqua M, Romano D (2003) Emissioni di benzene in Italia dal 1990 al 2000. Rapporti X/2003. APAT-Dipartimento Stato dell'Ambiente, Controlli e Sistemi Informativi-Unita Interdipartimentale Censimento Fonti di Emissione.
    [50] Council Directive 93/12/EEC of 23 March 1993 relating to the sulphur content of certain liquid fuels and Council Directive 98/70/EC of the European Parliament and of the Council of 13 October 1998 relating to the quality of petrol and diesel fuels and amending Council Directive 93/12/EEC.
    [51] Rappenglück B, Dasgupta PK, Leuchner M, et al. (2010) Formaldehyde and its relation to CO, PAN, and SO2 in the Houston-Galveston airshed. Atmos Chem Phys 10: 2413-2424. doi: 10.5194/acp-10-2413-2010
    [52] Movassaghi K, Russo MV, Avino P (2012) The determination and role of peroxyacetil nitrate in photochemical processes in atmosphere. Chem Central J 6: S8. doi: 10.1186/1752-153X-6-S2-S8
    [53] Parrish DD, Ryerson TB, Mellqvist J, et al. (2012) Primary and secondary sources of formaldehyde in urban atmospheres: Houston Texas region. Atmos Chem Phys 12: 3273-3288. doi: 10.5194/acp-12-3273-2012
    [54] Febo A, Perrino C, Bruno P, et al. (1999) Formation and occurrence of nitrous acid in the atmosphere (FORMONA), Final report to the European Commission (ENV4-CT95-0055). Consiglio Nazionale delle Ricerche, Rome.
    [55] Czader BH, Rappenglück B, Percell P (2012) Modeling nitrous acid and its impact on ozone and hydroxyl radical during the Texas Air Quality Study 2006. Atmos Chem Phys 12: 6939-6951. doi: 10.5194/acp-12-6939-2012
    [56] Wiesen P (2002) Nitrous acid and its influence on the oxidation capacity of the atmosphere (NITROCAT), final EU project report (EVK2-CT-1999-00025).
    [57] Acker K, Möller D, Auel R, et al. (2005) Concentrations of nitrous acid, nitric acid, nitrite and nitrate in the gas and aerosol phase at a site in the emission zone during ECOMPTE 2001 experiment. Atmos Res 74: 507-524. doi: 10.1016/j.atmosres.2004.04.009
    [58] Acker K, Febo A, Trick, et al. (2006) Nitrous acid in the urban area of Rome. Atmos Environ 40: 3123-3133. doi: 10.1016/j.atmosenv.2006.01.028
    [59] Heintzenberg J (1982) Size-segregated measurements of particulate elemental carbon and aerosol loght absorption at remote locations. Atmos Environ 16: 2461-2469. doi: 10.1016/0004-6981(82)90136-6
    [60] Avino P, Brocco D, Pareti S, Scalisi G (2003) Description of the carbonaceous particulate matter evolution in an urban area. Ann Chim 93: 21-26.
    [61] Avino P, Manigrasso M, Rosada A, et al. (2015) Measurement of organic and elemental carbon in downtown Rome and background area: physical behavior and chemical speciation. Environ Sci Process Impacts 17: 300-315. doi: 10.1039/C4EM00471J
    [62] Kourtidis KA, Ziomasa IC, Rappenglück B, et al. (1999) Evaporative traffic hydrocarbon emissions, traffic CO and speciated HC traffic emissions from the city of Athens. Atmos Environ 33: 3831-3842. doi: 10.1016/S1352-2310(98)00395-1
    [63] Leuchner M, Rappenglück B (2010) VOC source-receptor relationships in Houston during TexAQS-II. Atmos Environ 44: 4056-4067. doi: 10.1016/j.atmosenv.2009.02.029
    [64] Carter WPL (1994) Development of ozone reactivity scales for volatile organic compounds. J Air Waste Manage 44: 881-899. doi: 10.1080/1073161X.1994.10467290
    [65] Derwent RG, Jenkin ME (1991) Hydrocarbons and the long-range transport of Ozone and PAN across Europe. Atmos Environ 25A: 1661-1678.
    [66] Blake DR, Rowland FS (1995) Urban leakage of liquefied petroleum gas and its impact on Mexico City air quality. Science 269: 953-956. doi: 10.1126/science.269.5226.953
    [67] Winkler J, Blank P, Glaser K, et al. (2002) Ground-based and airborne measurements of nonmethane hydrocarbons in BERLIOZ: analysis and selected results. J Atmos Chem 42: 465-492. doi: 10.1023/A:1015709214016
    [68] Brocco D, Fratarcangeli R, Lepore L, et al. (1997) Determination of aromatic hydrocarbons in urban air of Rome. Atmos Environ 21: 557-566.
    [69] Rappenglück B, Fabian P (1999) Nonmethane hydrocarbons (NMHC) in the Greater Munich Area/Germany. Atmos Environ 33: 3843-3857. doi: 10.1016/S1352-2310(98)00394-X
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