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Sample records for acid vesicle ph

  1. Basis of antimalarial action: non-weak base effects of chloroquine on acid vesicle pH

    SciTech Connect

    Krogstad, D.J.; Schlesinger, P.H.

    1987-03-01

    Biologically active concentrations of chloroquine increase the pH of the parasite's acid vesicles within 3-5 min. This increase in pH results from two mechanisms, one of which is markedly reduced in chloroquine-resistant parasites. Because chloroquine is a weak base, it increases vesicle pH by that mechanism in chloroquine-susceptible and resistant parasites and mammalian cells (based on its two pKs and on the delta pH between the acid vesicle and the extracellular environment). In chloroquine-susceptible parasites, but not resistant parasites or mammalian cells, chloroquine increases the pH of acid vesicles 700- to 800-fold more than can be accounted for by its properties as a weak base. The increase in acid vesicle pH caused by these non-weak base effects of nanomolar chloroquine in susceptible parasites suggests that chloroquine acts by interfering with acid vesicle functions in the parasite such as the endocytosis and proteolysis of hemoglobin, and the intracellular targeting of lysosomal enzymes. The non-weak base effects of nanomolar chloroquine on parasite vesicle pH are also responsible for its safety because these chloroquine concentrations do not affect mammalian cells.

  2. Formation, stability, and pH sensitivity of free-floating, giant unilamellar vesicles using palmitic acid-cholesterol mixtures.

    PubMed

    Cottenye, Nicolas; Carbajal, Gustavo; Cui, Zhong-Kai; Ducharme, Philippe Dauphin; Mauzeroll, Janine; Lafleur, Michel

    2014-09-14

    Despite the fact that palmitic acid (PA) and cholesterol (Chol) do not form fluid bilayers once hydrated individually, giant unilamellar vesicles (GUVs) were formed from a mixture of palmitic acid and cholesterol, 30/70 mol/mol. These free-floating GUVs were stable over weeks, did not aggregate and were shown to be highly stable in alkaline pH compared to conventional phospholipid-based GUVs. Acidic pH-triggered payload release from the GUVs was associated with the protonation state of palmitic acid that dictated the mixing lipid properties, thus affecting the stability of the fluid lamellar phase. The successful formation of PA-Chol GUVs reveals the possibility to create monoalkylated amphiphile-based GUVs with distinct pH stability/sensitivity.

  3. Vesicles of 2-ketooctanoic acid in water.

    PubMed

    Xu, Huifang; Du, Na; Song, Yawen; Song, Shue; Hou, Wanguo

    2017-03-15

    We report the spontaneous formation of vesicles from 2-ketooctanoic acid (KOCOOH), a single-tailed weakly acidic surfactant, in water. The vesicles were characterized using negative-staining, cryogenic transmission electron microscopy, conductivity, and atomic force microscopy. The pH effect on the vesicle formation and the stability of the vesicular structures were determined. The vesicles form at a very low concentration (ca. 1.4 mM) and within a wide pH range (ca. 2-10). Uni- and multilamellar vesicle structures are observed, which coexist in the KOCOOH solution. The hydrogen bonding between KOCOOH molecules probably plays an important role in the formation of the vesicles. Importantly, the vesicles exhibit remarkable stability upon long-term storage, and in artificial seawater. KOCOOH vesicles are a good alternative model system for protocell-like vesicles, as they are easily formed under plausible prebiotic conditions. In addition, they may have the same potential applications, such as in medicine, chemical engineering, and biotechnology, as conventional vesicles. To the best of our knowledge, this is the first report on the vesicles of single-tailed keto-acid amphiphiles.

  4. Antimalarials increase vesicle pH in Plasmodium falciparum

    PubMed Central

    1985-01-01

    The asexual erythrocytic stage of the malarial parasite ingests and degrades the hemoglobin of its host red cell. To study this process, we labeled the cytoplasm of uninfected red cells with fluorescein-dextran, infected those cells with trophozoite- and schizont-rich cultures of Plasmodium falciparum, and harvested them 110-120 h later in the trophozoite stage. After lysis of the red cell cytoplasm with digitonin, the only fluorescence remaining was in small (0.5-0.9 micron) vesicles similar to the parasite's food vacuole. As measured by spectrofluorimetry, the pH of these vesicles was acid (initial pH 5.2- 5.4), and they responded to MgATP with acidification and to weak bases such as NH4Cl with alkalinization. These three properties are similar to those obtained with human fibroblasts and suggest that the endocytic vesicles of plasmodia are similar to those of mammalian cells. Each of the antimalarials tested (chloroquine, quinine, and mefloquine) as well as NH4Cl inhibited parasite growth at concentrations virtually identical to those that increased parasite vesicle pH. These results suggest two conclusions: (a) The increases in vesicle pH that we have observed in our digitonin-treated parasite preparation occur at similar concentrations of weak bases and antimalarials in cultures of parasitized erythrocytes, and (b) P. falciparum parasites are exquisitely dependent on vesicle pH during their asexual erythrocytic cycle, perhaps for processes analogous to endocytosis and proteolysis in mammalian cells, and that antimalarials and NH4Cl may act by interfering with these events. PMID:3905824

  5. Vesicles protect activated acetic acid.

    PubMed

    Todd, Zoe R; House, Christopher H

    2014-10-01

    Abstract Methyl thioacetate, or activated acetic acid, has been proposed to be central to the origin of life and an important energy currency molecule in early cellular evolution. We have investigated the hydrolysis of methyl thioacetate under various conditions. Its uncatalyzed rate of hydrolysis is about 3 orders of magnitude faster (K=0.00663 s(-1); 100°C, pH 7.5, concentration=0.33 mM) than published rates for its catalyzed production, making it unlikely to accumulate under prebiotic conditions. However, our experiments showed that methyl thioacetate was protected from hydrolysis when inside its own hydrophobic droplets. Further, we found that methyl thioacetate protection from hydrolysis was also possible in droplets of hexane and in the membranes of nonanoic acid vesicles. Thus, the hydrophobic regions of prebiotic vesicles and early cell membranes could have offered a refuge for this energetic molecule, increasing its lifetime in close proximity to the reactions for which it would be needed. This model of early energy storage evokes an additional critical function for the earliest cell membranes.

  6. Transfer of Oleic Acid between Albumin and Phospholipid Vesicles

    NASA Astrophysics Data System (ADS)

    Hamilton, James A.; Cistola, David P.

    1986-01-01

    The net transfer of oleic acid between egg phosphatidylcholine unilamellar vesicles and bovine serum albumin has been monitored by 13C NMR spectroscopy and 90% isotopically substituted [1-13C]oleic acid. The carboxyl chemical shifts of oleic acid bound to albumin were different from those for oleic acid in phospholipid vesicles. Therefore, in mixtures of donor particles (vesicles or albumin with oleic acid) and acceptor particles (fatty acid-free albumin or vesicles), the equilibrium distribution of oleic acid was determined from chemical shift and peak intensity data without separation of donor and acceptor particles. In a system containing equal masses of albumin and phospholipid and a stoichiometry of 4-5 mol of oleic acid per mol of albumin, the oleic acid distribution was pH dependent, with >= 80% of the oleic acid associated with albumin at pH 7.4; association was >= 90% at pH 8.0. Decreasing the pH below 7.4 markedly decreased the proportion of fatty acid bound to albumin; at pH 5.4, <= 10% of the oleic acid was bound to albumin and >90% was associated with vesicles. The distribution was reversible with pH and was independent of whether vesicles or albumin acted as a donor. These data suggest that pH may strongly influence the partitioning of fatty acid between cellular membranes and albumin. The 13C NMR method is also advantageous because it provides information about the structural environments of oleic acid bound to albumin or phospholipid, the ionization state of oleic acid in each environment, and the structural integrity of the vesicles. In addition, minimum and maximum limits for the exchange rates of oleic acid among different environments were obtained from the NMR data.

  7. Transfer of oleic acid between albumin and phospholipid vesicles

    SciTech Connect

    Hamilton, J.A.; Cistola, D.P.

    1986-01-01

    The net transfer of oleic acid between egg phosphatidylcholine unilamellar vesicles and bovine serum albumin has been monitored by TC NMR spectroscopy and 90% isotopically substituted (1- TC)oleic acid. The carboxyl chemical shifts of oleic acid bound to albumin were different from those for oleic acid in phospholipid vesicles. Therefore, in mixtures of donor particles, the equilibrium distribution of oleic acid was determined from chemical shift and peak intensity data without separation of donor and acceptor particles. In a system containing equal masses of albumin and phospholipid and a stoichiometry of 4-5 mol of oleic acid per mol of albumin, the oleic acid distribution was pH dependent, with greater than or equal to80% of the oleic acid associated with albumin at pH 7.4; association was greater than or equal to90% at pH 8.0. Decreasing the pH below 7.4 markedly decreased the proportion of fatty acid bound to albumin. The distribution was reversible with pH and was independent of whether vesicles or albumin acted as a donor. These data suggest that pH may strongly influence the partitioning of fatty acid between cellular membranes and albumin. The TC NMR method is also advantageous because it provides information about the structural environments of oleic acid bound to albumin or phospholipid, the ionization state of oleic acid in each environment, and the structural integrity of the vesicles. In addition, minimum and maximum limits for the exchange rates of oleic acid among different environments were obtained from the NMR data.

  8. Presynaptic pH and vesicle fusion in Drosophila larvae neurones.

    PubMed

    Caldwell, Lesley; Harries, Peter; Sydlik, Sebastian; Schwiening, Christof J

    2013-11-01

    Both intracellular pH (pHi) and synaptic cleft pH change during neuronal activity yet little is known about how these pH shifts might affect synaptic transmission by influencing vesicle fusion. To address this we imaged pH- and Ca(2+) -sensitive fluorescent indicators (HPTS, Oregon green) in boutons at neuromuscular junctions. Electrical stimulation of motor nerves evoked presynaptic Ca(2+) i rises and pHi falls (∼0.1 pH units) followed by recovery of both Ca(2+) i and pHi. The plasma-membrane calcium ATPase (PMCA) inhibitor, 5(6)-carboxyeosin diacetate, slowed both the calcium recovery and the acidification. To investigate a possible calcium-independent role for the pHi shifts in modulating vesicle fusion we recorded post-synaptic miniature end-plate potential (mEPP) and current (mEPC) frequency in Ca(2+) -free solution. Acidification by propionate superfusion, NH(4)(+) withdrawal, or the inhibition of acid extrusion on the Na(+)/H(+) exchanger (NHE) induced a rise in miniature frequency. Furthermore, the inhibition of acid extrusion enhanced the rise induced by propionate addition and NH(4)(+) removal. In the presence of NH(4)(+), 10 out of 23 cells showed, after a delay, one or more rises in miniature frequency. These findings suggest that Ca(2+) -dependent pHi shifts, caused by the PMCA and regulated by NHE, may stimulate vesicle release. Furthermore, in the presence of membrane permeant buffers, exocytosed acid or its equivalents may enhance release through positive feedback. This hitherto neglected pH signalling, and the potential feedback role of vesicular acid, could explain some important neuronal excitability changes associated with altered pH and its buffering.

  9. Presynaptic pH and vesicle fusion in Drosophila larvae neurones

    PubMed Central

    Caldwell, Lesley; Harries, Peter; Sydlik, Sebastian; Schwiening, Christof J

    2014-01-01

    Both intracellular pH (pHi) and synaptic cleft pH change during neuronal activity yet little is known about how these pH shifts might affect synaptic transmission by influencing vesicle fusion. To address this we imaged pH- and Ca2+-sensitive fluorescent indicators (HPTS, Oregon green) in boutons at neuromuscular junctions. Electrical stimulation of motor nerves evoked presynaptic Ca2+i rises and pHi falls (∼0.1 pH units) followed by recovery of both Ca2+i and pHi. The plasma-membrane calcium ATPase (PMCA) inhibitor, 5(6)-carboxyeosin diacetate, slowed both the calcium recovery and the acidification. To investigate a possible calcium-independent role for the pHi shifts in modulating vesicle fusion we recorded post-synaptic miniature end-plate potential (mEPP) and current (mEPC) frequency in Ca2+-free solution. Acidification by propionate superfusion, NH4+ withdrawal, or the inhibition of acid extrusion on the Na+/H+ exchanger (NHE) induced a rise in miniature frequency. Furthermore, the inhibition of acid extrusion enhanced the rise induced by propionate addition and NH4+ removal. In the presence of NH4+, 10 out of 23 cells showed, after a delay, one or more rises in miniature frequency. These findings suggest that Ca2+-dependent pHi shifts, caused by the PMCA and regulated by NHE, may stimulate vesicle release. Furthermore, in the presence of membrane permeant buffers, exocytosed acid or its equivalents may enhance release through positive feedback. This hitherto neglected pH signalling, and the potential feedback role of vesicular acid, could explain some important neuronal excitability changes associated with altered pH and its buffering. Synapse 67:729–740, 2013. PMID:23649934

  10. pH response of carboxy-terminated colorimetric polydiacetylene vesicles.

    PubMed

    Kew, Simon J; Hall, Elizabeth A H

    2006-04-01

    Carboxy-terminated polydiacetylene vesicles are known to undergo dramatic color transitions in response to exposure to external stimuli such as pH, temperature, and receptor-ligand binding. FTIR spectroscopy was used to identify the breakdown in the interfacial hydrogen-bonding interactions of the carboxylic acid headgroups of polymerized 10,12-tricosadiynoic acid (TRCDA) vesicles in aqueous solution during pH chromic transition. The headgroup structure was monitored as the chromic transition takes place and the dissociation dependence of the pKa was determined. Due to the attenuated acidity of the interfacially confined carboxy groups, which exhibit pKa values in the range 9.5-9.9, it was found that the deprotonation-triggered blue-red chromic transition occurred in the pH range 9.0-10.1 and that the mechanism of the transition required interaction with the surface carboxyl group, which is of importance in the design of a biochromic mechanism using PDA assemblies. Transmission electron microscopy and FTIR spectroscopy revealed that the surface ionization and the pH-induced chromogenic transition was also accompanied by a dramatic vesicle-planar morphological transition alongside subtle changes to the alkyl chain conformation and packing. A two-step mechanism was implicated as causing the chromic transition that first involves surface deprotonation and then specific cation binding, which can aid the design of sensitive surface-ligand chemistry for new PDA structures.

  11. B/Ca in coccoliths and relationship to calcification vesicle pH and dissolved inorganic carbon concentrations

    NASA Astrophysics Data System (ADS)

    Stoll, Heather; Langer, Gerald; Shimizu, Nobumichi; Kanamaru, Kinuyo

    2012-03-01

    Coccolithophorid algae are microscopic but prolific calcifiers in modern and ancient oceans. When the pH of seawater is modified, as may occur in the future due to ocean acidification, different species and strains of coccolithophorids have exhibited diverse calcification responses in laboratory culture. Since their biomineralization is a completely intracellular process, it is unclear why their response should be affected by extracellular seawater pH. Variations in the B/Ca in coccoliths are potential indicators of pH shifts in the intracellular coccolith vesicle where calcification occurs, because B/Ca in abiogenic calcites increases at higher pH due to the greater abundance of borate ions, the only B species incorporated into calcite. We used a SIMS ion probe to measure B/Ca of coccoliths from three different strains of Emiliania huxleyi and one strain of Coccolithus braarudii braarudii cultured under different seawater pH conditions to ascertain if the B/Ca can be used to elucidate how coccolithophorids respond to changing ocean pH. These data are interpreted with the aid of a conceptual model of cellular boron acquisition by coccolithophorids. Based on uptake in other plants, we infer that boron uptake by coccolithophorid cells is dominated by passive uptake of boric acid across the lipid bilayer. Subsequently, in the alkaline coccolith vesicle (C.V.), boron speciates according to the C.V. pH, and borate is incorporated into the coccolith. At increasing seawater pH, the relative abundance of the neutral boric acid in seawater decreases, lowering the potential B flux into the cell. Homeostasis or constant pH of the coccolith vesicle results in a decrease of the B/Ca in the coccolith with increasing seawater pH. In contrast, if coccolith vesicle pH increases with increasing seawater pH, then the B/Ca will increase as the fraction of borate in the coccolith vesicle increases. The coccolith B/Ca is also expected to depend inversely on the dissolved inorganic

  12. pH-responsive jello: gelatin gels containing fatty acid vesicles.

    PubMed

    Dowling, Matthew B; Lee, Jae-Ho; Raghavan, Srinivasa R

    2009-08-04

    We describe a new way to impart pH-responsive properties to gels of biopolymers such as gelatin. This approach involves the embedding of pH-sensitive nanosized vesicles within the gel. The vesicles employed here are those of sodium oleate (NaOA), a fatty-acid-based amphiphile with a single C18 tail. In aqueous solution, NaOA undergoes a transition from vesicles at a pH approximately 8 to micelles at a pH higher than approximately 10. Here, we combine NaOA and gelatin at pH 8.3 to create a vesicle-loaded gel and then bring the gel in contact with a pH 10 buffer solution. As the buffer diffuses into the gel, the vesicles within the gel get transformed into micelles. Accordingly, a vesicle-micelle front moves through the gel, and this can be visually identified by the difference in turbidity between the two regions. Vesicle disruption can also be done in a spatially selective manner to create micelle-rich domains within a vesicle-loaded gel. A possible application of the above approach is in the area of pH-dependent controlled release. A vesicle-to-micelle transition releases hydrophilic solutes encapsulated within the vesicles into the bulk gel, and in turn these solutes can rapidly diffuse out of the gel into the external bath. Experiments with calcein dye confirm this concept and show that we can indeed use the pH in the bath to tune the release rate of solutes from vesicle-loaded gels.

  13. pH-responsive poly(methacrylic acid)-grafted hollow silica vesicles.

    PubMed

    Lay, Chee Leng; Tan, Hui Ru; Lu, Xuehong; Liu, Ye

    2011-02-18

    Poly(methacrylic acid)-grafted hollow silica vesicles (PMAA-g-hollow silica vesicles) were obtained through a grafting-from approach. PMAA brushes were formed by performing atom-transfer radical polymerisation of sodium methacrylate with an initiator attached to the hollow silica spheres. PMAA-g-hollow silica vesicles were characterised by using TEM, thermogravimetric analysis (TGA) and FTIR spectroscopy. pH-dependent ξ potential and (1)H NMR spectra of PMAA-g-hollow silica vesicles were measured, and the results indicated that MAA brushes in PMAA-g-hollow silica vesicles had a lower ionisation degree and low solubility in acidic aqueous solution, for example, pH 3.4, but a higher ionisation degree and high solubility when the pH was higher than 7. Also it was demonstrated that calcein blue and fluorescein isothiocyanate (FITC) labelled dextran (M(n):10 kDa) could be encapsulated in the interiors of the PMAA-g-hollow silica vesicles with a negligible amount in PMAA brushes at pH 2, and pH-triggered release of calcein blue and FITC-labelled dextran from PMAA-g-hollow silica vesicles was observed at pH 7.4. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Role of a transmembrane pH gradient in epinephrine transport by chromaffin granule membrane vesicles.

    PubMed

    Schuldiner, S; Fishkes, H; Kanner, B I

    1978-08-01

    ATP-driven transport and accumulation of epinephrine in chromaffin granule membrane vesicles isolated from bovine adrenal medulla is inhibited by the proton ionophores carbonylcyanide p-trifluoromethoxyphenylhydrazone and nigericin, but not by valinomycin. Moreover, an artificially imposed pH gradient (interior acid) is able to drive this reserpine-sensitive transport system in the absence of ATP. Dicyclohexylcarbodiimide, an inactivator of the chromaffin granule membrane-bound ATPase, completely inhibits ATP-dependent epinephrine accumulation, but has much less effect when an imposed pH gradient is the driving force for epinephrine transport. The findings provide a strong indication that a pH gradient (interior acid) is the immediate driving force for epinephrine uptake in these storage granules and suggest that ATP-driven epinephrine transport is the result of two processes: (i) generation of a proton electrochemical gradient (interior acid and positive) by the membrane-bound, proton-translocating ATPase; and (ii) pH gradient-driven accumulation of the catecholamine.

  15. Role of a transmembrane pH gradient in epinephrine transport by chromaffin granule membrane vesicles.

    PubMed Central

    Schuldiner, S; Fishkes, H; Kanner, B I

    1978-01-01

    ATP-driven transport and accumulation of epinephrine in chromaffin granule membrane vesicles isolated from bovine adrenal medulla is inhibited by the proton ionophores carbonylcyanide p-trifluoromethoxyphenylhydrazone and nigericin, but not by valinomycin. Moreover, an artificially imposed pH gradient (interior acid) is able to drive this reserpine-sensitive transport system in the absence of ATP. Dicyclohexylcarbodiimide, an inactivator of the chromaffin granule membrane-bound ATPase, completely inhibits ATP-dependent epinephrine accumulation, but has much less effect when an imposed pH gradient is the driving force for epinephrine transport. The findings provide a strong indication that a pH gradient (interior acid) is the immediate driving force for epinephrine uptake in these storage granules and suggest that ATP-driven epinephrine transport is the result of two processes: (i) generation of a proton electrochemical gradient (interior acid and positive) by the membrane-bound, proton-translocating ATPase; and (ii) pH gradient-driven accumulation of the catecholamine. PMID:29292

  16. Active auxin uptake by zucchini membrane vesicles: quantitation using ESR volume and delta pH determinations

    SciTech Connect

    Lomax, T.L.; Mehlhorn, R.J.; Briggs, W.R.

    1985-10-01

    Closed and pH-tight membrane vesicles prepared from hypocotyls of 5-day-old dark-grown seedlings of Cucurbita pepo accumulate the plant growth hormone indole-3-acetic acid along an imposed proton gradient (pH low outside, high inside). The use of electron paramagnetic spin probes permitted quantitation both of apparent vesicle volume and magnitude of the pH gradient. Under the experimental conditions used, hormone accumulation was at minimum 20-fold, a value 4 times larger than what one would predict if accumulation reflected only diffusional equilibrium at the measured pH gradient. It is concluded that hormone uptake is an active process, with each protonated molecule of hormone accompanied by an additional proton. Experiments with ionophores confirm that it is the pH gradient itself which drives the uptake.

  17. Lipid interaction of Pseudomonas aeruginosa exotoxin A. Acid-triggered permeabilization and aggregation of lipid vesicles.

    PubMed Central

    Menestrina, G; Pederzolli, C; Forti, S; Gambale, F

    1991-01-01

    We have investigated the interaction of Pseudomonas exotoxin A with small unilamellar vesicles comprised of different phospholipids as a function of pH, toxin, and lipid concentration. We have found that this toxin induces vesicle permeabilization, as measured by the release of a fluorescent dye. Permeabilization is due to the formation of ion-conductive channels which we have directly observed in planar lipid bilayers. The toxin also produces vesicle aggregation, as indicated by an increase of the turbidity. Aggregation and permeabilization have completely different time course and extent upon toxin dose and lipid composition, thus suggesting that they are two independent events. Both time constants decrease by lowering the pH of the bulk phase or by introducing a negative lipid into the vesicles. Our results indicate that at least three steps are involved in the interaction of Pseudomonas exotoxin A with lipid vesicles. After protonation of one charged group the toxin becomes competent to bind to the surface of the vesicles. Binding is probably initiated by an electrostatic interaction because it is absolutely dependent on the presence of acidic phospholipids. Binding is a prerequisite for the subsequent insertion of the toxin into the lipid bilayer, with a special preference for phosphatidylglycerol-containing membranes, to form ionic channels. At high toxin and vesicle concentrations, bound toxin may also induce aggregation of the vesicles, particularly when phosphatidic acid is present in the lipid mixture. A quenching of the intrinsic tryptophan fluorescence of the protein, which is induced by lowering the pH of the solution, becomes more drastic in the presence of lipid vesicles. However, this further quenching takes so long that it cannot be a prerequisite to either vesicle permeabilization or aggregation. Pseudomonas exotoxin A shares many of these properties with other bacterial toxins like diphtheria and tetanus toxin. Images FIGURE 7 FIGURE 8 FIGURE 12

  18. A new model system for lipid interactions in stratum corneum vesicles: effects of lipid composition, calcium, and pH.

    PubMed

    Hatfield, R M; Fung, L W

    1999-01-12

    We prepared large unilamellar vesicles (LUVs) with three different stratum corneum lipid compositions: constant amounts of ceramides (55 wt %) and fatty acids (15%) with varying amounts of cholesterol sulfate (0-15%) and cholesterol (15-30%). One of the compositions served as a model for normal stratum corneum, while the second one served as a model for recessive X-linked ichthyosis stratum corneum. The third composition consisted of no cholesterol sulfate. Intervesicle lipid interactions in these LUVs were monitored by fluorescence methods for content leakage, and contents mixing at pH 9, in the absence and presence of Ca2+, and at pH 6. Since the content leakage and contents mixing assays were originally developed for phospholipid vesicles, we characterized the probe binding and the probe quenching properties for stratum corneum LUV systems, and modified the assays slightly accordingly. The time-dependent fluorescence intensity changes in the probe-containing LUVs at pH 9 and 6 and in response to the addition of calcium were monitored. Our results demonstrated that all three types of LUVs were relatively stable at pH 9. Addition of Ca2+ or decreasing the pH to 6 activated intervesicle lipid mixing followed by vesicle fusion and lysis. We found that the LUVs with no cholesterol sulfate and 30% cholesterol exhibited a more extensive Ca2+- or low-pH-activated intervesicle lipid interaction than LUVs with either 5% cholesterol sulfate and 25% cholesterol or 15% cholesterol sulfate and 15% cholesterol. These results suggest that fusogenic agents such as Ca2+ and H+ act to neutralize the fatty acids in the lipid bilayer of stratum corneum vesicles. The inclusion of 5-15% cholesterol sulfate helps to prevent the collapse of fused vesicles into other structures.

  19. Influence of molecular weight and pH on adsorption of chitosan at the surface of large and giant vesicles.

    PubMed

    Quemeneur, Francois; Rinaudo, Marguerite; Pépin-Donat, Brigitte

    2008-01-01

    This paper describes the mechanisms of adsorption of chitosan, a positively charged polyelectrolyte, on the DOPC lipid membrane of large and giant unilamellar vesicles (respectively, LUVs and GUVs). We observe that the variation of the zeta potential of LUVs as a function of chitosan concentration is independent on the chitosan molecular weight (Mw). This result is interpreted in terms of electrostatic interactions, which induce a flat adsorption of the chitosan on the surface of the membrane. The role of electrostatic interactions is further studied by observing the variation of the zeta potential as a function of the chitosan concentration for two different charge densities tuned by the pH. Results show a stronger chitosan-membrane affinity at pH 6 (lipids are negatively charged, and 40% chitosan amino groups are protonated) than at pH 3.4 (100% of protonated amino groups but zwitterionic lipids are positively charged) which confirms that adsorption is of electrostatic origin. Then, we investigate the stability of decorated LUVs and GUVs in a large range of pH (6.0 < pH < 12.0) in order to complete a previous study made in acidic conditions [Quemeneur et al. Biomacromolecules 2007, 8, 2512-2519]. A comparative study of the variation of the zeta potential as a function of the pH (2.0 < pH < 12.0) reveals a difference in behavior between naked and chitosan-decorated LUVs. This result is further confirmed by a comparative observation by optical microscopy of naked and chitosan-decorated GUVs in basic conditions (6.0 < pH < 12.0): at pH > 10.0, in the absence of chitosan, the vesicles present complex shapes, contrary to the chitosan-decorated vesicles which remain spherical, confirming thus that chitosan remains adsorbed on vesicles in basic conditions up to pH = 12.0. These results, in addition with our previous data, show that the chitosan-decorated vesicles are stable over a very broad range of pH (2.0 < pH < 12.0), which holds promise for their in vivo

  20. Model Systems of Precursor Cellular Membranes: Long-Chain Alcohols Stabilize Spontaneously Formed Oleic Acid Vesicles

    PubMed Central

    Rendón, Adela; Carton, David Gil; Sot, Jesús; García-Pacios, Marcos; Montes, Ruth; Valle, Mikel; Arrondo, José-Luis R.; Goñi, Felix M.; Ruiz-Mirazo, Kepa

    2012-01-01

    Oleic acid vesicles have been used as model systems to study the properties of membranes that could be the evolutionary precursors of more complex, stable, and impermeable phospholipid biomembranes. Pure fatty acid vesicles in general show high sensitivity to ionic strength and pH variation, but there is growing evidence that this lack of stability can be counterbalanced through mixtures with other amphiphilic or surfactant compounds. Here, we present a systematic experimental analysis of the oleic acid system and explore the spontaneous formation of vesicles under different conditions, as well as the effects that alcohols and alkanes may have in the process. Our results support the hypothesis that alcohols (in particular 10- to 14-C-atom alcohols) contribute to the stability of oleic acid vesicles under a wider range of experimental conditions. Moreover, studies of mixed oleic-acid-alkane and oleic-acid-alcohol systems using infrared spectroscopy and Langmuir trough measurements indicate that precisely those alcohols that increased vesicle stability also decreased the mobility of oleic acid polar headgroups, as well as the area/molecule of lipid. PMID:22339864

  1. A confocal study on the visualization of chromaffin cell secretory vesicles with fluorescent targeted probes and acidic dyes.

    PubMed

    Moreno, Alfredo; SantoDomingo, Jaime; Fonteriz, Rosalba I; Lobatón, Carmen D; Montero, Mayte; Alvarez, Javier

    2010-12-01

    Secretory vesicles have low pH and have been classically identified as those labelled by a series of acidic fluorescent dyes such as acridine orange or neutral red, which accumulate into the vesicles according to the pH gradient. More recently, several fusion proteins containing enhanced green fluorescent protein (EGFP) and targeted to the secretory vesicles have been engineered. Both targeted fluorescent proteins and acidic dyes have been used, separately or combined, to monitor the dynamics of secretory vesicle movements and their fusion with the plasma membrane. We have now investigated in detail the degree of colocalization of both types of probes using several fusion proteins targeted to the vesicles (synaptobrevin2-EGFP, Cromogranin A-EGFP and neuropeptide Y-EGFP) and several acidic dyes (acridine orange, neutral red and lysotracker red) in chromaffin cells, PC12 cells and GH(3) cells. We find that all the acidic dyes labelled the same population of vesicles. However, that population was largely different from the one labelled by the targeted proteins, with very little colocalization among them, in all the cell types studied. Our data show that the vesicles containing the proteins more characteristic of the secretory vesicles are not labelled by the acidic dyes, and vice versa. Peptide glycyl-L-phenylalanine 2-naphthylamide (GPN) produced a rapid and selective disruption of the vesicles labelled by acidic dyes, suggesting that they could be mainly lysosomes. Therefore, these labelling techniques distinguish two clearly different sets of acidic vesicles in neuroendocrine cells. This finding should be taken into account whenever vesicle dynamics is studied using these techniques.

  2. Acid loading test (pH)

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003615.htm Acid loading test (pH) To use the sharing features on this page, please enable JavaScript. The acid loading test (pH) measures the ability of the ...

  3. Ultrasound and pH Dually Responsive Polymer Vesicles for Anticancer Drug Delivery

    NASA Astrophysics Data System (ADS)

    Chen, Wenqin; Du, Jianzhong

    2013-07-01

    Recently, smart polymer vesicles have attracted increasing interest due to their endless potential applications such as tunable delivery vehicles for the treatment of degenerative diseases. However, the evolution of stimuli-responsive vesicles from bench to bedside still seems far away for the limitations of current stimuli forms such as temperature, light, redox, etc. Since ultrasound combined with chemotherapy has been widely used in tumor treatment and the pH in tumor tissues is relatively low, we designed herein a novel polymer vesicle that respond to both physical (ultrasound) and chemical (pH) stimuli based on a PEO-b-P(DEA-stat-TMA) block copolymer, where PEO is short for poly(ethylene oxide), DEA for 2-(diethylamino)ethyl methacrylate and TMA for (2-tetrahydrofuranyloxy)ethyl methacrylate. These dually responsive vesicles show noncytotoxicity below 250 μg/mL and can encapsulate anticancer drugs, exhibiting retarded release profile and controllable release rate when subjected to ultrasound radiation or varying pH in tris buffer at 37°C.

  4. Ultrasound and pH Dually Responsive Polymer Vesicles for Anticancer Drug Delivery

    PubMed Central

    Chen, Wenqin; Du, Jianzhong

    2013-01-01

    Recently, smart polymer vesicles have attracted increasing interest due to their endless potential applications such as tunable delivery vehicles for the treatment of degenerative diseases. However, the evolution of stimuli-responsive vesicles from bench to bedside still seems far away for the limitations of current stimuli forms such as temperature, light, redox, etc. Since ultrasound combined with chemotherapy has been widely used in tumor treatment and the pH in tumor tissues is relatively low, we designed herein a novel polymer vesicle that respond to both physical (ultrasound) and chemical (pH) stimuli based on a PEO-b-P(DEA-stat-TMA) block copolymer, where PEO is short for poly(ethylene oxide), DEA for 2-(diethylamino)ethyl methacrylate and TMA for (2-tetrahydrofuranyloxy)ethyl methacrylate. These dually responsive vesicles show noncytotoxicity below 250 μg/mL and can encapsulate anticancer drugs, exhibiting retarded release profile and controllable release rate when subjected to ultrasound radiation or varying pH in tris buffer at 37°C. PMID:23831819

  5. Citrate uptake into tonoplast vesicles from acid lime (Citrus aurantifolia) juice cells.

    PubMed

    Brune, A; Gonzalez, P; Goren, R; Zehavi, U; Echeverria, E

    1998-12-01

    Citrate transport into the vacuoles of acid lime juice cells was investigated using isolated tonoplast vesicles. ATP stimulated citrate uptake in the presence or in the absence of a Delta mu H+. Energization of the vesicles only by an artificial K+ gradient (establishing an inside-positive Delta psi) also resulted in citrate uptake as was the case of a Delta pH dominated Delta mu H+. Addition of inhibitors to endomembrane ATPases showed no direct correlation between the inhibition to the tonoplast bound H+/ATPase and citrate uptake. The data indicated that, although some citrate uptake can be accounted for by Delta psi and by a direct primary active transport mechanism involving ATP, under in vivo conditions of vacuolar pH of 2.0, citrate uptake is driven by Delta pH.

  6. Glutamate transport into synaptic vesicles. Roles of membrane potential, pH gradient, and intravesicular pH.

    PubMed

    Tabb, J S; Kish, P E; Van Dyke, R; Ueda, T

    1992-08-05

    Glutamate, the major excitatory neurotransmitter in the mammalian central nervous system, is transported into bovine synaptic vesicles in a manner that is ATP dependent and requires a vesicular electrochemical proton gradient. We studied the electrical and chemical elements of this driving force and evaluated the effects of chloride on transport. Increasing concentrations of Cl- were found to increase the steady-state ATP-dependent vesicular pH gradient (delta pH) and were found to concomitantly decrease the vesicular membrane potential (delta psi). Low millimolar chloride concentrations, which cause 3-6-fold stimulation of vesicular glutamate uptake, caused small but measurable increases in delta pH and decreases in delta psi, when compared to control vesicles in the absence of chloride. Nigericin in potassium buffers was used to alter the relative proportions of delta pH and delta psi. Compared to controls, at all chloride concentrations tested, nigericin virtually abolished delta pH and increased the vesicle interior positive delta psi. Concomitantly, nigericin increased ATP-dependent glutamate uptake in 0-1 mM chloride but decreased glutamate uptake in 4 mM (45%), 20 mM (80%), and 140 mM (75%) Cl- (where delta pH in the absence of nigericin was large). These findings suggest that either delta psi, delta pH, or a combination can drive glutamate uptake, but to different degrees. In the presence of 4 mM Cl-, where uptake is optimal, both delta psi and delta pH contribute to the driving force for uptake. When the extravesicular pH was increased from 7.4 to 8.0, more Cl- was required to stimulate vesicular glutamate uptake. In the absence of Cl-, as extravesicular pH was lowered to 6.8, uptake was over 3-fold greater than it was at pH 7.4. As extravesicular pH was reduced from 8.0 toward 6.8, less Cl- was required for maximal stimulation. Decreasing the extravesicular pH from 8.0 to 6.8 in the absence of Cl- significantly increased glutamate uptake activity, even

  7. Self-Assembly of Bilayer Vesicles Made of Saturated Long Chain Fatty Acids.

    PubMed

    Douliez, Jean-Paul; Houssou, Bérénice Houinsou; Fameau, A-Laure; Navailles, Laurence; Nallet, Frédéric; Grélard, Axelle; Dufourc, Erick J; Gaillard, Cédric

    2016-01-19

    Saturated long chain fatty acids (sLCFA, e.g., C14:0, C16:0, and C18:0) are potentially the greenest and cheapest surfactants naturally available. However, because aqueous sodium soaps of sLCFA are known to crystallize, the self-assembly of stable bilayer vesicles has not been reported yet. Here, by using such soaps in combination with guanidine hydrochloride (GuHCl), which has been shown recently to prevent crystallization, we were capable of producing stable bilayer vesicles made of sLCFA. The phase diagrams were established for a variety of systems showing that vesicles can form in a broad range of composition and pH. Both solid state NMR and small-angle neutron scattering allowed demonstrating that in such vesicles sLCFA are arranged in a bilayer structure which exhibits similar dynamic and structural properties as those of phospholipid membranes. We expect these vesicles to be of interest as model systems of protocells and minimal cells but also for various applications since fatty acids are potentially substitutes to phospholipids, synthetic surfactants, and polymers.

  8. Sodium Chloride Triggered the Fusion of Vesicle Composed of Fatty Acid Modified Protic Ionic Liquid: A New Insight into the Membrane Fusion Monitored through Fluorescence Lifetime Imaging Microscopy.

    PubMed

    Kundu, Niloy; Banerjee, Pavel; Kundu, Sangita; Dutta, Rupam; Sarkar, Nilmoni

    2017-01-12

    The development of stable vesicular assemblies and the understanding of their interaction and dynamics in aqueous solution are long-standing topics in the research of chemistry and biology. Fatty acids are known to form vesicle structure in aqueous solution depending on the pH of the medium. Protic ionic liquid of fatty acid with ethyl amine (oleate ethyl amine, OEA) as a component spontaneously forms a vesicle in aqueous solution. The general comparison of dynamics and interaction of these two vesicles have been drawn using fluorescence correlation spectroscopy (FCS) and fluorescence lifetime imaging microscopy (FLIM) measurements. Further, FLIM images of a single vesicle are taken at multiple wavelengths, and the solvation of the probe molecules has been observed from the multiwavelength FLIM images. The lifetime of the probe molecule in OEA vesicle is higher than that in simple fatty acid vesicles. Therefore, it suggests that the membrane of the OEA vesicle is more dehydrated compared to that of fatty acid vesicles, and it facilitates OEA vesicles to fuse themselves in the presence of electrolyte, sodium chloride (NaCl). However, under the same conditions, only fatty acid vesicles do not fuse. The fusion of OEA vesicles is successfully demonstrated by the time scan FLIM measurements. The different events in the fusion process are analyzed in the light of the reported model of vesicle fusion. Finally, the local viscosity of the water pool of the vesicle is determined using kiton red, as a molecular rotor. With addition of NaCl, the fluidity in the interior of the vesicle is increased which leads to disassembly of vesicle. The rich dynamic properties of this vesicular assembly and the FLIM based approach of vesicle fusion will provide better insight into the growth of a protocell membrane.

  9. Acid Rain, pH & Acidity: A Common Misinterpretation.

    ERIC Educational Resources Information Center

    Clark, David B.; Thompson, Ronald E.

    1989-01-01

    Illustrates the basis for misleading statements about the relationship between pH and acid content in acid rain. Explains why pH cannot be used as a measure of acidity for rain or any other solution. Suggests that teachers present acidity and pH as two separate and distinct concepts. (RT)

  10. Acid Rain, pH & Acidity: A Common Misinterpretation.

    ERIC Educational Resources Information Center

    Clark, David B.; Thompson, Ronald E.

    1989-01-01

    Illustrates the basis for misleading statements about the relationship between pH and acid content in acid rain. Explains why pH cannot be used as a measure of acidity for rain or any other solution. Suggests that teachers present acidity and pH as two separate and distinct concepts. (RT)

  11. Reconciling Ligase Ribozyme Activity with Fatty Acid Vesicle Stability

    PubMed Central

    Anella, Fabrizio; Danelon, Christophe

    2014-01-01

    The “RNA world” and the “Lipid world” theories for the origin of cellular life are often considered incompatible due to the differences in the environmental conditions at which they can emerge. One obstacle resides in the conflicting requirements for divalent metal ions, in particular Mg2+, with respect to optimal ribozyme activity, fatty acid vesicle stability and protection against RNA strand cleavage. Here, we report on the activity of a short L1 ligase ribozyme in the presence of myristoleic acid (MA) vesicles at varying concentrations of Mg2+. The ligation rate is significantly lower at low-Mg2+ conditions. However, the loss of activity is overcompensated by the increased stability of RNA leading to a larger amount of intact ligated substrate after long reaction periods. Combining RNA ligation assays with fatty acid vesicles we found that MA vesicles made of 5 mM amphiphile are stable and do not impair ligase ribozyme activity in the presence of approximately 2 mM Mg2+. These results provide a scenario in which catalytic RNA and primordial membrane assembly can coexist in the same environment. PMID:25513761

  12. pH and Titratable Acidity

    NASA Astrophysics Data System (ADS)

    Sadler, George D.; Murphy, Patricia A.

    There are two interrelated concepts in food analysis that deal with acidity: pH and titratable acidity. Each of these quantities is analytically determined in separate ways and each has its own particular impact on food quality. Titratable acidity deals with measurement of the total acid concentration contained within a food (also called total acidity). This quantity is determined by exhaustive titration of intrinsic acids with a standard base. Titratable acidity is a better predictor of acid's impact on flavor than pH.

  13. Decrease of pH gradients in tonoplast vesicles by NO/sub 3//sup -/ and Cl/sup -/: evidence for H/sup +/-coupled anion transport

    SciTech Connect

    Schumaker, K.S.; Sze, H.

    1987-03-01

    Chloride or nitrate decreased a pH gradient (measured as (/sup 14/C)methylamine accumulation) in tonoplast-enriched vesicles. The ..delta..pH decrease was dependent on the anion concentration. These effects are independent of the anion-sensitive H/sup +/-ATPase of the tonoplast, since the pH gradient (acid inside) was imposed artificially using a pH jump or a K/sup +/ gradient and nigericin. 4,4'-Diisothiocyano-2,2'-stilbene disulfonic acid partially prevented the decrease in pH gradient induced by Cl/sup -/. Two possible models to account for this anion-dependent decrease of ..delta..pH are: (a) H/sup +/ loss is accompanied by Cl/sup -/ or NO/sub 3//sup -/ efflux from the vesicles via H/sup +//anion symport systems on the tonoplast and (b) H/sup +/ loss is accompanied by Cl/sup -/ or NO/sub 3//sup -/ uptake into the vesicles via H/sup +//anion antiport systems. Depending on the requirements and conditions of the cell, these two systems would serve to either mobilize Cl/sup -/ and NO/sub 3//sup -/ stored in the vacuole for use in the cytoplasm or to drive anions into the vacuole. Chloride or nitrate also decreased a pH gradient in fractions containing plasma membrane and Golgi, implying that these membranes may have similar H/sup +/-coupled anion transport systems.

  14. Vesicles

    MedlinePlus

    ... the top of a pin or up to 5 millimeters wide. A larger blister is called a bulla. In many cases, vesicles break easily and release their fluid onto the skin. When this fluid dries, yellow crusts may remain on the skin surface.

  15. Large and giant vesicles "decorated" with chitosan: effects of pH, salt or glucose stress, and surface adhesion.

    PubMed

    Quemeneur, Francois; Rammal, Ayman; Rinaudo, Marguerite; Pépin-Donat, Brigitte

    2007-08-01

    This paper describes the behavior of large and giant unilamellar vesicles (LUVs and GUVs, respectively) in the presence of chitosan, a positively charged polyelectrolyte. Variation of the zeta-potential of LUVs as a function of chitosan concentration is studied for two different molecular weights (MW) after a preliminary study devoted to pH and salt effects on zeta-potential in order to discriminate among the effects of protons, salt, and chitosan concentrations. The difference observed between pH and salt effects on the one hand and chitosan on the other allows us to conclude there is a strong LUV-chitosan interaction. In presence of chitosan, the zeta-potential of LUVs becomes positive and two distinct regimes of variation are suggested and interpreted as follows: a first step consists of chitosan adsorption flat on the membrane (independent of MW) followed by a possible reorganization of the polymer of higher molecular weight on the surface, giving rise to loops. Then a comparative observation of the effect of pH and salt by optical microscopy is made on naked and chitosan-decorated GUVs. Results further confirm a membrane-chitosan interaction and are interpreted in the light of the results obtained for LUVs in terms of both electrostatic and hydrophobic interaction. A large majority of decorated vesicles remain stable down to pH = 1 while in the absence of chitosan they burst quickly at pH between 2 and 3. Osmotic pressure and net charge change due to addition of HCl results in a decrease in the diameter of the decorated vesicles, which remain spherical while forming tubes of lipids. In presence of NaCl, a higher resistance of decorated vesicles is also evidenced (they are stable for NaCl concentrations up to 10-1 M while naked vesicles burst when [NaCl] is between 10-2 and 10-3 M). At higher salt concentration, aggregation of decorated vesicles occurs, which is attributed to the screening of electrostatic repulsions between vesicles covered by the positively

  16. In vitro fusion of endocytic vesicles: effects of reagents that alter endosomal pH.

    PubMed

    Pless, D D; Wellner, R B

    1996-07-01

    Ricin, a plant toxin that binds to galactose-terminated glycoproteins and glycolipids on the cell surface, is internalized into endosomes before reaching the cytosol where it exerts its toxic activity. Fusion of early endosomes containing ricin or transferrin was demonstrated by using postnuclear supernatant fractions from K-562 cells. For both ligands, fusion depended on time, temperature, and ATP and was blocked by preincubation with N-ethylmaleimide. Some reagents that increase endosomal pH, the ionophores monensin and nigericin and the weak base chloroquine, stimulated the rate of fusion. However, bafilomycin A1, a specific inhibitor of vacuolar H(+)-ATPases, did not alter the rate of fusion. Moreover, it reduced or eliminated stimulation caused by monensin, nigericin, or chloroquine. Thus, the increased rate of fusion did not correlate with the higher lumenal pH of the endosome. The results suggest instead that fusion was stimulated by reagents that promoted accumulation of cations within the vesicles.

  17. Fatty acid vesicles acting as expanding horizon for transdermal delivery.

    PubMed

    Kumar, Lalit; Verma, Shivani; Kumar, Sanjeev; Prasad, Deo Nandan; Jain, Amit Kumar

    2017-03-01

    The body is protected against the external environment by the skin due to its physical barrier nature. Stratum corneum composed of corneocytes surrounded by lipid region performs a major barrier function as it lies in the uppermost area of skin. Alteration in barrier function, increase in permeability, and disorganization of stratum corneum represent diseased skin. Drugs applied to the diseased skin should induce a local effect at the site of application or area close to it along with cutaneous absorption rather than percutaneous absorption. Conventional formulations like ointments, gels, and creams suffer from the drawback of limited local activity. For the enhancement of drug penetration and localization of the drug at the site of action approaches explored are liposomes, niosomes, ethosomes microparticles, and solid lipid nanoparticles. Vesicles composed of fatty acids like oleic acid and linoleic acid represent the new approach used for transdermal penetration and localization. In this review article, our major aim was to explore the applications of fatty acid vesicles for transdermal delivery of various bioactives.

  18. Self-Propelled Oil Droplets and Their Morphological Change to Giant Vesicles Induced by a Surfactant Solution at Low pH.

    PubMed

    Banno, Taisuke; Tanaka, Yuki; Asakura, Kouichi; Toyota, Taro

    2016-09-20

    Unique dynamics using inanimate molecular assemblies based on soft matter have drawn much attention for demonstrating far-from-equilibrium chemical systems. However, there are no soft matter systems that exhibit a possible pathway linking the self-propelled oil droplets to formation of giant vesicles stimulated by low pH. In this study, we conceived an experimental oil-in-water emulsion system in which flocculated particles composed of a imine-containing oil transformed to spherical oil droplets that self-propelled and, after coming to rest, formed membranous figures. Finally, these figures became giant vesicles. From NMR, pH curves, and surface tension measurements, we determined that this far-from-equilibrium phenomenon was due to the acidic hydrolysis of the oil, which produced a benzaldehyde derivative as an oil component and a primary amine as a surfactant precursor, and the dynamic behavior of the hydrolytic products in the emulsion system. These findings afforded us a potential linkage between mobile droplet-based protocells and vesicle-based protocells stimulated by low pH.

  19. Nuclear magnetic resonance study of acetic acid permeation of large unilamellar vesicle membranes.

    PubMed Central

    Alger, J R; Prestegard, J H

    1979-01-01

    The permeation of acetic acid through large unilamellar phospholipid vesicle membranes has been investigated using the unique capability of nuclear magnetic resonance to characterize flow under pseudo-equilibrium conditions. Two types of experiments have been employed: total line shape analysis and selective population transfer. These techniques are sensitive to permeation on time scales ranging form 0.001 to 10.0 s. The permeation rate dependence on pH and acetic acid concentration indicates that the neutral acetic acid monomer is the dominant permeant species with a permeation coefficient of 5 +/- 2 x 10-4 cm/s. Mechanisms of permeation and the applicability of nuclear magnetic resonance methodology are discussed. PMID:262441

  20. Study of vesicle size distribution dependence on pH value based on nanopore resistive pulse method

    NASA Astrophysics Data System (ADS)

    Lin, Yuqing; Rudzevich, Yauheni; Wearne, Adam; Lumpkin, Daniel; Morales, Joselyn; Nemec, Kathleen; Tatulian, Suren; Lupan, Oleg; Chow, Lee

    2013-03-01

    Vesicles are low-micron to sub-micron spheres formed by a lipid bilayer shell and serve as potential vehicles for drug delivery. The size of vesicle is proposed to be one of the instrumental variables affecting delivery efficiency since the size is correlated to factors like circulation and residence time in blood, the rate for cell endocytosis, and efficiency in cell targeting. In this work, we demonstrate accessible and reliable detection and size distribution measurement employing a glass nanopore device based on the resistive pulse method. This novel method enables us to investigate the size distribution dependence of pH difference across the membrane of vesicles with very small sample volume and rapid speed. This provides useful information for optimizing the efficiency of drug delivery in a pH sensitive environment.

  1. Floret-shaped solid domains on giant fluid lipid vesicles induced by pH

    NASA Astrophysics Data System (ADS)

    Sofou, Stavroula; Bandekar, Amey

    2012-02-01

    Lateral lipid phase separation and domain formation induced by changes in pH is significant in liposome-based drug delivery: environmentally responsive lipid heterogeneities can be tuned to alter collective membrane properties such as drug release and drug carrier reactivity impacting, therefore, the therapeutic outcomes. At the micron-meter scale, fluorescence microscopy on Giant Unilamellar fluid Vesicles (GUVs) shows that lowering pH (from 7.0 to 5.0) promotes the condensation of titratable PS or PA lipids into beautiful floret-shaped solid domains in which lipids are tightly packed via H-bonding and VdWs interactions. Solid domains phenomenologically comprise a circular ``core'' cap beyond which interfacial instabilities emerge resembling leaf-like stripes of almost vanishing Gaussian curvature independent of GUVs' preparation path and in agreement with a general condensation mechanism. Increasing incompressibility of domains is strongly correlated with larger number of thinner stripes per domain, and increasing relative rigidity of domains with smaller core cap areas. Line tension drives domain ripening, however the final domain shape is a result of enhanced incompressibility and rigidity maximized by domain coupling across the bilayer. Introduction of a transmembrane osmotic gradient (hyperosmotic on the outer lipid leaflet) allows the domain condensation process to reach its maximum extent which, however, is limited by the minimal expansivity of the continuous fluid membrane.

  2. A molecular beacon-based DNA switch for reversible pH sensing in vesicles and live cells.

    PubMed

    Narayanaswamy, Nagarjun; Nair, Raji R; Suseela, Y V; Saini, Deepak Kumar; Govindaraju, T

    2016-07-05

    In this Communication, a molecular beacon-based DNA switch (LMB) is developed as an efficient and reversible pH sensing probe. Remarkably, LMB exhibited reversible structural transition between the closed (molecular beacon) and open (A-motif) states very efficiently in synthetic vesicles and live cells without the need for any transfection agents.

  3. Vesicles from pH-regulated reversible gemini amino-acid surfactants as nanocapsules for delivery.

    PubMed

    Lv, Jing; Qiao, Weihong; Li, Zongshi

    2016-10-01

    Reversible transition from micelles to vesicles by regulating pH were realized by gemini amino-acid surfactants N,N'-dialkyl-N,N'-diacetate ethylenediamine. Measurement results of ζ-potential at different pH and DLS at varying solvents revealed that the protonation between H(+) and double NCH2COO(-) groups (generating NH(+)CH2COO(-)), expressed as pKa1 and pKa2, is the key driving force to control the aggregation behaviors of gemini surfactant molecule. Effect of pH on the bilayer structure was studied in detail by using steady-state fluorescence spectroscopy of hydrophobic pyrene and Coumarin 153 (C153) respectively and fluorescence resonance energy transfer (FRET) from C153 to Rhodamine 6G (R6G). Various pH-regulated and pH-reversible self-assemblies were obtained in one surfactant system. Vitamin D3 was encapsulated in vesicle bilayers to form nano-VD3-capsules as VD3 supplement agent for health care products. By using the electrostatic attraction between Ca(2+) and double -COO(-) groups, nano-VD3-capsules with Ca(2+) coated outermost layers were prepared as a formulation for VD3 and calcium co-supplement agent. DLS and TEM were performed to check stability and morphology of the nano-capsules. It is concluded that the pH-regulated gemini amino-acid surfactants can be used to construct colloidal systems for delivering hydrophobic drugs or nutritions without lipids at human physiological pH level. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Growth and instability of a phospholipid vesicle in a bath of fatty acids

    NASA Astrophysics Data System (ADS)

    Dervaux, J.; Noireaux, V.; Libchaber, A. J.

    2017-06-01

    Using a microfluidic trap, we study the behavior of individual phospholipid vesicles in contact with fatty acids. We show that spontaneous fatty acids insertion inside the bilayer is controlled by the vesicle size, osmotic pressure difference across the membrane and fatty acids concentration in the external bath. Depending on these parameters, vesicles can grow spherically or become unstable and fragment into several daughter vesicles. We establish the phase diagram for vesicle growth and we derive a simple thermodynamic model that reproduces the time evolution of the vesicle volume. Finally, we show that stable growth can be achieved on an artificial cell expressing a simple set of bacterial cytoskeletal proteins, paving the way toward artificial cell reproduction.

  5. pH gradient effects on chloride transport across basolateral membrane vesicles from guinea-pig jejunum.

    PubMed Central

    Touzani, K; Alvarado, F; Vasseur, M

    1997-01-01

    1. The effects of alkaline-inside pH gradients on 36Cl- uptake were quantified by using brush-border membrane (BBM) and basolateral membrane (BLM) vesicles from guinea-pig jejunum. 2. With BBM vesicles, a pHo/pHi gradient of 5.0/7.5 yielded fast overshoots involving a random, non-obligatory Cl(-)-H+ symport, strongly inhibited by CCCP. In contrast, BLM vesicles responded to similar pH gradients with much smaller, delayed overshoots, unaffected by CCCP. 3. The initial Cl- entry rates into BLM vesicles were a function of each pHo, pHi and delta pH value. They were stimulated by valinomycin in the presence of inward-directed K+ gradients. Short-circuiting the membrane potential with equilibrated K+ and valinomycin inhibited pH gradient-dependent Cl- uptake, but only partially. 4. Taken together, these results indicate that guinea-pig jejunal BLM vesicles possess both Cl- conductance and Cl(-)-H+ symport activities. 5. Even when different, the BBM and the BLM symporters are mechanistically similar. Neither of them involves a Cl(-)-OH- antiport, nor a simultaneous Cl(-)-anion exchange mechanism. Rather, for each membrane, all of these activities (symport, anion exchange) can be explained in terms of a single mobile carrier acting as a random, non-obligatory Cl(-)-H+ symporter where exchange occurs simply by counterflow. Net Cl- translocation via either the ternary (Cl(-)-C-H+) or the binary (Cl(-)-C) complexes accounts, respectively, for the existence of two, operationally distinct, electroneutral and rheogenic components. 6. The BBM symporter appears to involve an AE2 protein, but the molecular identity of the BLM one remains to be established. PMID:9147326

  6. Facile preparation of a cationic poly(amino acid) vesicle for potential drug and gene co-delivery

    NASA Astrophysics Data System (ADS)

    Ding, Jianxun; Xiao, Chunsheng; He, Chaoliang; Li, Mingqiang; Li, Di; Zhuang, Xiuli; Chen, Xuesi

    2011-12-01

    A novel pH-responsive poly(amino acid) grafted with oligocation was prepared through the combination of ring-opening polymerization (ROP) and subsequent atom transfer radical polymerization (ATRP). Firstly, poly(γ-2-chloroethyl-L-glutamate) (PCELG) with a pendent 2-chloroethyl group was synthesized through ROP of γ-2-chloroethyl-L-glutamate N-carboxyanhydride (CELG NCA) using n-hexylamine as the initiator. Then, PCELG was used to initiate the ARTP of 2-aminoethyl methacrylate hydrochloride (AMA), yielding poly(L-glutamate)-graft-oligo(2-aminoethyl methacrylate hydrochloride) (PLG-g-OAMA). The pKa of PLG-g-OAMA was 7.3 established by the acid-base titration method. The amphiphilic poly(amino acid) could directly self-assemble into a vesicle in PBS. The vesicle was characterized by TEM and DLS. Hydrophilic DOX·HCl was loaded into the hollow core of the vesicle. The in vitro release behavior of DOX·HCl from the vesicle in PBS could be adjusted by the solution pH. In vitro cell experiments revealed that the vesicle could reduce the toxicity of the DOX·HCl. In addition, the preliminary gel retardation assay displayed that PLG-g-OAMA could efficiently bind DNA at a PLG-g-OAMA/DNA weight ratio of 0.3 or above, indicating its potential use as a gene carrier. More in-depth studies of the PLG-g-OAMA vesicle for drug and gene co-delivery in vitro and in vivo are in progress.

  7. Effects of internal and external pH on amiloride-blockable Na transport across toad urinary bladder vesicles

    SciTech Connect

    Garty, H.; Civan, E.D.; Civan, M.M.

    1985-01-01

    The authors have examined the effect of internal and external pH on Na+ transport across toad bladder membrane vesicles. Of the total SSNa uptake measured 0.5-2.0 min after introducing tracer, 80 +/- 4% (mean +/- SE, n = 9) is blocked by the diuretic with a KI of 2 X 10(-8) M. Thus, this amiloride-sensitive flux is mediated by the apical sodium-selective channels. Varying the internal (cytosolic) pH over the physiologic range 7.0-8.0 had no effect on sodium transport; this result suggests that variation of intracellular pH in vivo has no direct apical effect on modulating sodium uptake. On the other hand, SSNa was directly and monotonically dependent on external pH. External acidification also reduced the amiloride-sensitive efflux across the walls of the vesicles. This inhibition of 22Na efflux was noted at external Na concentrations of both 0.2 microM and 53 mM. These results are different from those reported with whole toad bladder. A number of possible bases for these differences are considered and discussed. They suggest that the natriferic response induced by mucosal acidification of whole toad urinary bladder appears to operate indirectly through one or more factors, presumably cytosolic, present in whole cells and absent from the vesicles.

  8. Interaction of tetanus toxin with lipid vesicles. Effects of pH, surface charge, and transmembrane potential on the kinetics of channel formation.

    PubMed Central

    Menestrina, G; Forti, S; Gambale, F

    1989-01-01

    We investigated the interaction of tetanus toxin with small unilamellar vesicles composed of different phospholipids as a function of pH, toxin concentration, temperature, and ionic strength of the solution. Tetanus toxin increased the permeability of the vesicles to fluorescent markers of molecular weight up to 700. The time course of the permeabilization was described as the sum of two exponential components of which the faster accounts for more than 70% of the total effect. Both time constants decreased when the pH of the solution was lowered and when vesicles contained negative lipids. These results can be explained in terms of a phenomenological model based on reaction rate theory. The model assumes that tetanus toxin, after equilibrating with the local pH existing at the surface of the vesicles, inserts into the lipid bilayer forming an ionic channel through which solutes can diffuse. Trigger event for the insertion of the toxin is the protonation, and consequent neutralization of one charged group which makes the molecule more hydrophobic. The intrinsic pK of this group was found to be 3.4 +/- 0.2, suggesting that it may be a carboxyl group. Since the toxin equilibrates with the local pH, the enhancing effect of acidic phospholipids is merely explained by the creation of a negative surface potential which increases the local proton concentration. This was confirmed by the inhibitory effect of high Na+ concentration which reduced the surface charge by screening and specific binding. We found still small differences between the lipids tested and the following order of sensitivity to the action of the toxin: phosphatidylinositol greater than phosphatidylserine greater than phosphatidylcholine approximately cholesterol. The activation energy for the two time constants was found to be 19.8 and 14.8 kcal/mol, fast and slow component, respectively, i.e., slightly larger than that for pure diffusion through the bilayer. The permeabilization induced by tetanus toxin

  9. The transmembrane pH gradient drives uphill folate transport in rabbit jejunum. Direct evidence for folate/hydroxyl exchange in brush border membrane vesicles.

    PubMed Central

    Schron, C M; Washington, C; Blitzer, B L

    1985-01-01

    In rabbit jejunal, but not ileal brush border membrane vesicles, an outwardly directed OH- gradient (pH 7.7 inside, pH 5.5 outside) markedly stimulated the initial velocity of folate (0.1 microM) uptake compared with uptake in the absence of a pH gradient. Under pH gradient conditions, folate was transiently accumulated at a concentration four times that found at equilibrium (over-shoot), implying uphill transport of the vitamin. Equilibrium folate uptake was inversely proportional to medium osmolality, suggesting uptake into an osmotically sensitive space. pH gradient-stimulated folate uptake was markedly reduced by inhibitors of anion exchange (4,4'-diisothiocyano-2,2'-disulfonic acid stilbene; 4-acetamido-4-isothiocyanostilbene-2,2'-disulfonic acid; furosemide), and was saturable (folate Km = 0.19 +/- 0.02 microM; Vmax = 12.8 +/- 0.4 pmol X mg protein-1 X min-1). Imposition of an inside-positive electrical potential did not stimulate folate uptake, suggesting that stimulation by a pH gradient was not due to an induced electrical potential. In contrast, an inwardly directed Na+ or K+ gradient did not stimulate folate uptake. These findings provide evidence for a carrier on the jejunal brush border membrane that mediates folate/OH- exchange (or H+/folate co-transport), and are consonant with the known presence of an outwardly directed OH- gradient in vivo (brush border acid microclimate), an acidic pH optimum for intestinal folate uptake, and the primary role of the jejunum in folate absorption. PMID:4056063

  10. Light-dependent delta pH and membrane potential changes in halobacterial vesicles coupled to sodium transport

    SciTech Connect

    Kamo, N.; Racanelli, T.; Packer, L.

    1982-01-01

    Bacteriorhodopsin and Halorhodopsin present in Halobacterium halobium strains have been investigated in relation to Na/sup +//H/sup +/ exchange in isolated cell envelope vesicles. Upon illumination, these retinal proteins result in extrusion of sodium ions by either an electrogenic Na/sup +//Ha/sup +/ antiporter and/or a direct sodium pump. Since a molecular characterization of these mechanism(s) of sodium extrusion has not yet been realized, it was of interest to measure directly the light- and sodium-dependent changes in delta pH and membrane potential under nearly identical conditions in S9 and R1mR cell membrane vesicles to gain information on the relation of these retinal proteins to sodium extrusion. These activities were evaluated in terms of their dependence on light intensity, and on the inhibitory effect of chemical modifiers of carboxyl groups (carbodiimides); electroneutral exchanges (monensin and triphenyltin); digitoxin and some analogues; and phloretin. Under most of the conditions and treatments employed, light- and sodium-dependent delta pH led to similar effects in both membrane vesicle types. Hence, it is concluded that the delta pH and delta psi which arise from sodium transport occur by either a single mechanism or by one which shares common features.

  11. Sequestration of Proteins by Fatty Acid Coacervates for Their Encapsulation within Vesicles.

    PubMed

    Garenne, David; Beven, Laure; Navailles, Laurence; Nallet, Frédéric; Dufourc, Erick J; Douliez, Jean-Paul

    2016-10-17

    Encapsulating biological materials in lipid vesicles is of interest for mimicking cells; however, except in some particular cases, such processes do not occur spontaneously. Herein, we developed a simple and robust method for encapsulating proteins in fatty acid vesicles in high yields. Fatty acid based, membrane-free coacervates spontaneously sequester proteins and can reversibly form membranous vesicles upon varying the pH value, the precrowding feature in coacervates allowing for protein encapsulation within vesicles. We then produced enzyme-enriched vesicles and show that enzymatic reactions can occur in these micrometric capsules. This work could be of interest in the field of synthetic biology for building microreactors. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Unique pH dynamics in GABAergic synaptic vesicles illuminates the mechanism and kinetics of GABA loading

    PubMed Central

    Egashira, Yoshihiro; Takase, Miki; Watanabe, Shoji; Ishida, Junji; Fukamizu, Akiyoshi; Kaneko, Ryosuke; Yanagawa, Yuchio; Takamori, Shigeo

    2016-01-01

    GABA acts as the major inhibitory neurotransmitter in the mammalian brain, shaping neuronal and circuit activity. For sustained synaptic transmission, synaptic vesicles (SVs) are required to be recycled and refilled with neurotransmitters using an H+ electrochemical gradient. However, neither the mechanism underlying vesicular GABA uptake nor the kinetics of GABA loading in living neurons have been fully elucidated. To characterize the process of GABA uptake into SVs in functional synapses, we monitored luminal pH of GABAergic SVs separately from that of excitatory glutamatergic SVs in cultured hippocampal neurons. By using a pH sensor optimal for the SV lumen, we found that GABAergic SVs exhibited an unexpectedly higher resting pH (∼6.4) than glutamatergic SVs (pH ∼5.8). Moreover, unlike glutamatergic SVs, GABAergic SVs displayed unique pH dynamics after endocytosis that involved initial overacidification and subsequent alkalization that restored their resting pH. GABAergic SVs that lacked the vesicular GABA transporter (VGAT) did not show the pH overshoot and acidified further to ∼6.0. Comparison of luminal pH dynamics in the presence or absence of VGAT showed that VGAT operates as a GABA/H+ exchanger, which is continuously required to offset GABA leakage. Furthermore, the kinetics of GABA transport was slower (τ > 20 s at physiological temperature) than that of glutamate uptake and may exceed the time required for reuse of exocytosed SVs, allowing reuse of incompletely filled vesicles in the presence of high demand for inhibitory transmission. PMID:27601664

  13. Co-delivery of doxorubicin and arsenite with reduction and pH dual-sensitive vesicle for synergistic cancer therapy

    NASA Astrophysics Data System (ADS)

    Zhang, Lu; Xiao, Hong; Li, Jingguo; Cheng, Du; Shuai, Xintao

    2016-06-01

    Drug resistance is the underlying cause for therapeutic failure in clinical cancer chemotherapy. A prodrug copolymer mPEG-PAsp(DIP-co-BZA-co-DOX) (PDBD) was synthesized and assembled into a nanoscale vesicle comprising a PEG corona, a reduction and pH dual-sensitive hydrophobic membrane and an aqueous lumen encapsulating doxorubicin hydrochloride (DOX.HCl) and arsenite (As). The dual stimulation-sensitive design of the vesicle gave rise to rapid release of the physically entrapped DOX.HCl and arsenite inside acidic lysosomes, and chemically conjugated DOX inside the cytosol with high glutathione (GSH) concentration. In the optimized concentration range, arsenite previously recognized as a promising anticancer agent from traditional Chinese medicine can down-regulate the expressions of anti-apoptotic and multidrug resistance proteins to sensitize cancer cells to chemotherapy. Consequently, the DOX-As-co-loaded vesicle demonstrated potent anticancer activity. Compared to the only DOX-loaded vesicle, the DOX-As-co-loaded one induced more than twice the apoptotic ratio of MCF-7/ADR breast cancer cells at a low As concentration (0.5 μM), due to the synergistic effects of DOX and As. The drug loading strategy integrating chemical conjugation and physical encapsulation in stimulation-sensitive carriers enabled efficient drug loading in the formulation.Drug resistance is the underlying cause for therapeutic failure in clinical cancer chemotherapy. A prodrug copolymer mPEG-PAsp(DIP-co-BZA-co-DOX) (PDBD) was synthesized and assembled into a nanoscale vesicle comprising a PEG corona, a reduction and pH dual-sensitive hydrophobic membrane and an aqueous lumen encapsulating doxorubicin hydrochloride (DOX.HCl) and arsenite (As). The dual stimulation-sensitive design of the vesicle gave rise to rapid release of the physically entrapped DOX.HCl and arsenite inside acidic lysosomes, and chemically conjugated DOX inside the cytosol with high glutathione (GSH) concentration. In the

  14. Evidence for an UDP-glucuronic acid/phenol glucuronide antiport in rat liver microsomal vesicles.

    PubMed

    Bánhegyi, G; Braun, L; Marcolongo, P; Csala, M; Fulceri, R; Mandl, J; Benedetti, A

    1996-04-01

    The transport of glucuronides synthesized in the luminal compartment of the endoplasmic reticulum by UDP-glucuronosyltransferase isoenzymes was studied in rat liver microsomal vesicles. Microsomal vesicles were loaded with p-nitrophenol glucuronide (5 mM), phenolphthalein glucuronide or UDP-glucuronic acid, by a freeze-thawing method. In was shown that: (i) the loading procedure resulted in millimolar intravesicular concentrations of the different loading compounds; (ii) addition of UDP-glucuronic acid (5 mM) to the vesicles released both intravesicular glucuronides within 1 min; (iii) glucuronides stimulated the release of UDP-glucuronic acid from UDP acid-loaded microsomal vesicles; (iv) trans-stimulation of UDP-glucuronic acid entry by loading of microsomal vesicles with p-nitrophenol glucuronide, phenolphthalein glucuronide, UDP-glucuronic acid and UDP-N-acetyl-glucosamine almost completely abolished the latency of UDP-glucuronosyltransferase, although mannose 6-phosphatase latency remained unaltered; (v) the loading compounds by themselves did not stimulate UDP-glucuronosyltransferase activity. This study indicates that glucuronides synthesized in the lumen of endoplasmic reticulum can leave by an antiport, which concurrently transports USP-glucuronic acid into the lumen of the endoplasmic reticulum.

  15. Nonrandom distribution of sialic acid over the cell surface of bristle- coated endocytic vesicles of the sinusoidal endothelium cells

    PubMed Central

    1978-01-01

    Previous studies with protein tracers have shown that the luminal surface of the vascular endothelium of the bone marrow is endocytic. The endocytosis occurs through the formation of large bristle-coated vesicles (LCV). The anionic charge distribution in this process was examined at the luminal surface of the endothelial cell, At pH 1.8, colloidal iron (CI), native ferritin, and polycationic ferritin (PCF) are bound by the luminal surface of the endothelial cell, but not at the sites of LCV formation. PCF used over a pH range of 1.8--7.2 (CI is unstable at higher pH levels) revealed LCV binding of this agent in increasing manner from pH 3.5 upwards. PCF binding at low pH (1.8) at the endothelial cell surface was markedly reduced by neuraminidase. Neuraminidase did not reduce PCF binding by the endothelial cell surface nor by the LCV at higher pH levels. It is concluded that the luminal surface of the endothelial cell has exposed sialic acid groups which are absent or significantly diminished at endocytic sites. The free surface of the endothelial cells as well as the sites of endocytosis have, in addition, anionic material with a pKa higher than that of sialic acid (pKa 2.6). These anionic materials may be different at the sites of endocytosis as compared to those present at the free cell surface. PMID:29050

  16. Self-assembled polyhydroxy fatty acids vesicles: a mechanism for plant cutin synthesis.

    PubMed

    Heredia-Guerrero, José A; Benítez, José J; Heredia, Antonio

    2008-03-01

    Despite its biological importance, the mechanism of formation of cutin, the polymeric matrix of plant cuticles, has not yet been fully clarified. Here, for the first time, we show the participation in the process of lipid vesicles formed by the self-assembly of endogenous polyhydroxy fatty acids. The accumulation and fusion of these vesicles (cutinsomes) at the outer part of epidermal cell wall is proposed as the mechanism for early cuticle formation.

  17. Smart pH- and reduction-dual-responsive folate-PEG-coated polymeric lipid vesicles for tumor-triggered targeted drug delivery

    NASA Astrophysics Data System (ADS)

    Wang, Sheng; Wang, Hanjie; Liu, Zhongyun; Wang, Liangliang; Wang, Xiaomin; Su, Lin; Chang, Jin

    2014-06-01

    To improve their therapeutic index, designed nanocarriers should preferentially accumulate in tumor tissues and then rapidly enter tumor cells to release the encapsulated drugs in a triggered manner. In this article, a new kind of a smart pH- and reduction-dual-responsive drug delivery system based on folate-PEG-coated polymeric lipid vesicles (FPPLVs) formed from amphiphilic dextran derivatives was designed and prepared successfully. PEG chains with pH-sensitive hydrazone bonds, stearyl alcohol (SA) chains with reduction-sensitive disulfide bonds and folate were connected to a dextran main chain. The newly developed FPPLVs had a nano-sized structure (~50 nm) with a PEG coating. The in vitro DOX release profiles showed that the FPPLVs achieved a triggered drug release in response to acidic pH and reducing environments due to the cleavage of hydrazone bonds and disulfide bonds. It has also been demonstrated by an in vitro cellular uptake study that the FPPLVs lose their PEG coating as well as expose the folate in acidic conditions, which allows them to efficiently enter tumor cells through ligand-receptor interactions. In vitro cytotoxicity measurements also confirmed that FPPLVs exhibited pronounced antitumor activity against HeLa cells. These results suggest that FPPLVs are promising carriers for smart antitumor drug delivery applications.To improve their therapeutic index, designed nanocarriers should preferentially accumulate in tumor tissues and then rapidly enter tumor cells to release the encapsulated drugs in a triggered manner. In this article, a new kind of a smart pH- and reduction-dual-responsive drug delivery system based on folate-PEG-coated polymeric lipid vesicles (FPPLVs) formed from amphiphilic dextran derivatives was designed and prepared successfully. PEG chains with pH-sensitive hydrazone bonds, stearyl alcohol (SA) chains with reduction-sensitive disulfide bonds and folate were connected to a dextran main chain. The newly developed FPPLVs had a

  18. Light-activated amino acid transport in Halobacterium halobium envelope vesicles

    NASA Technical Reports Server (NTRS)

    Macdonald, R. E.; Lanyi, J. K.

    1977-01-01

    Vesicles prepared from Halobacterium halobium cell envelopes accumulate amino acids in response to light-induced electrical and chemical gradients. Nineteen of 20 commonly occurring amino acids have been shown to be actively accumulated by these vesicles in response to illumination or in response to an artificially created Na+ gradient. On the basis of shared common carriers the transport systems can be divided into eight classes, each responsible for the transport of one or several amino acids: arginine, lysine, histidine; asparagine, glutamine; alanine, glycine, threonine, serine; leucine, valine, isoleucine, methionine; phenylalanine, tyrosine, tryptophan; aspartate; glutamate; proline. Available evidence suggests that these carriers are symmetrical in that amino acids can be transported equally well in both directions across the vesicle membranes. A tentative working model to account for these observations is presented.

  19. Light-activated amino acid transport in Halobacterium halobium envelope vesicles

    NASA Technical Reports Server (NTRS)

    Macdonald, R. E.; Lanyi, J. K.

    1977-01-01

    Vesicles prepared from Halobacterium halobium cell envelopes accumulate amino acids in response to light-induced electrical and chemical gradients. Nineteen of 20 commonly occurring amino acids have been shown to be actively accumulated by these vesicles in response to illumination or in response to an artificially created Na+ gradient. On the basis of shared common carriers the transport systems can be divided into eight classes, each responsible for the transport of one or several amino acids: arginine, lysine, histidine; asparagine, glutamine; alanine, glycine, threonine, serine; leucine, valine, isoleucine, methionine; phenylalanine, tyrosine, tryptophan; aspartate; glutamate; proline. Available evidence suggests that these carriers are symmetrical in that amino acids can be transported equally well in both directions across the vesicle membranes. A tentative working model to account for these observations is presented.

  20. Prebiotic oligomerization of amino acids inside lipid vesicles of unsaturated and saturated fatty acids in hydrothermal environments

    NASA Astrophysics Data System (ADS)

    Imai, E.; Furuuchi, R.; Nemoto, A.; Hatori, K.; Honda, H.; Matsuno, K.

    We have already attempted an experimental model simulating seawater circulation in the vicinity of hydrothermal vents in the primitive ocean. We used a flow reactor that was constructed for simulating the pressure and temperature conditions of the hydrothermal vents. In the flow reactor, a high-temperature high-pressure fluid at 125˜ 250°C and at 20MPa was injected into a low temperature (0˜ 40°C ) chamber that was maintained at about the same high pressure as the fluid. We then experimentally examined a possibility of oligomerizing amino acids on or inside lipid vesicles. We compared three different kinds of lipid vesicles made of unsaturated fatty acids (oleic acid), saturated fatty acids (decanoic acid) and phospholipids (DPPC). Identification of the oligomeric products was made with the aid of an HPLC analysis. The oligomeric yields from glycine increased significantly in the presence of lipid vesicles compared to the case of their absence. On the other hand, there was found no significant difference in the yields of oligomers between in the presence of lipid vesicles dissolved by surfactant and in their absence. The possibility of lipid molecules serving as catalysts for oligomerization may be dismissed. The diameters of those lipid vesicles observed under a phase contrast microscope were about 10 micrometer or less. The total volume shared by oleic acid vesicles was about 5 % of the total volume of the suspension. Oligomerization of glycine inside oleic acid vesicles was enhanced more than 15 times compared to that proceeding in their outside. Enhancement of oligomerization of glycine in the presence of lipid vesicles was repeated as the reactants revisited the interface zone between the hot and cold regions. Even those lipid vesicles made of saturated fatty acid such as decanoic acid could have been functional in enhancing the oligomerization of monomers in their inside in the primitive ocean. References E. Imai, et al. (1999) Science 283, 831-833. H

  1. pH [Measure of Acidity].

    ERIC Educational Resources Information Center

    Henderson, Paula

    This autoinstructional program deals with the study of the pH of given substances by using litmus and hydrion papers. It is a learning activity directed toward low achievers involved in the study of biology at the secondary school level. The time suggested for the unit is 25-30 minutes (plus additional time for further pH testing). The equipment…

  2. Dissipation of pH gradients in tonoplast vesicles and liposomes by mixtures of acridine orange and anions. [Avena sativa L. ; Beta vulgaris

    SciTech Connect

    Pope, A.J.; Leigh, R.A. )

    1988-04-01

    Acridine orange altered the response to anions of both ATP and inorganic pyrophosphate-dependent pH gradient formation in tonoplast vesicles isolated from oat (Avena sativa L.) roots and red beet (Beta vulgaris L.) storage tissue. When used as a fluorescent pH probe in the presence of I{sup {minus}}, ClO{sub 3}{sup {minus}}, NO{sub 3}{sup {minus}}, Br{sup {minus}}, or SCN{sup {minus}}, acridine orange reported lower pH gradients than either quinacrine or ({sup 14}C)methylamine. Acridine orange, but not quinacrine, reduced ({sup 14}C)methylamine accumulation when NO{sub 3}{sup {minus}} was present indicating that the effect was due to a real decrease in the size of the pH gradient, not a misreporting of the gradient by acridine orange. Other experiments indicated that acridine orange and NO{sub 3}{sup {minus}} increased the rate of pH gradient collapse both in tonoplast vesicles and in liposomes of phosphatidylcholine and that the effect in tonoplast vesicles was greater at 24{degree}C than at 12{degree}C. It is suggested that acridine orange and certain anions increase the permeability of membranes to H{sup +}, possibly because protonated acridine orange and the anions form a lipophilic ion pair within the vesicle which diffuses across the membrane thus discharging the pH gradient.

  3. Pluronic P85/poly(lactic acid) vesicles as novel carrier for oral insulin delivery.

    PubMed

    Xiong, Xiang Yuan; Li, Qi Han; Li, Yu Ping; Guo, Liang; Li, Zi Ling; Gong, Yan Chun

    2013-11-01

    Poly(lactic acid)-b-Pluronic-b-poly(lactic acid) (PLA-P85-PLA) vesicles were developed as novel carrier for oral insulin delivery. PLA-P85-PLA block copolymer was synthesized by ring opening polymerization of the monomer l-lactide using Pluronic copolymer P85 as the initiator. Insulin-loaded PLA-P85-PLA vesicles were prepared by dialysis method and the mean diameter of insulin-loaded PLA-P85-PLA vesicles was determined to be 178 nm. The cytotoxicity studies using human ovarian cancer cells OVCAR-3 indicate that PLA-P85-PLA block copolymer has good biocompatibility. Both in vitro and in vivo release behavior of insulin loaded in PLA-P85-PLA vesicles were studied. It was observed that insulin was released out gradually from PLA-P85-PLA vesicles and almost all insulin was released out 7.5h later. More importantly, for the oral administration of insulin-loaded PLA-P85-PLA vesicles at insulin doses of 200 IU/kg, the minimum blood glucose concentration was observed in the diabetic mice test after 2.5h, which was 15% of initial glucose level. Furthermore, the blood glucose concentration increased slowly to 31.8% of initial blood glucose concentration after 10.5h and was maintained at this level for at least an additional 14h (32.5% of initial blood glucose concentration at 24.5h). These results proved that PLA-P85-PLA vesicles could be promising polymeric carriers for oral insulin delivery application due to their sustained and enhanced hypoglycemic effect. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Effect of surface and membrane potentials on IAA (indoleactic acid) uptake and binding by zucchini membrane vesicles

    SciTech Connect

    Clark, K.A.; Goldsmith, M.H.M.

    1986-08-01

    The polar transport of the endogenous hormone controlling extension growth of plant cells, indoleacetic acid (IAA), is thought to depend on transmembrane pH and electrical gradients resulting in part from the action of proton ATPases in the plasma membrane. Elements of this transport process are permeation of the membrane by the undissociated lipophilic indoleacetic acid (IAAH) from the acidic apoplast, followed by dissociation of the weak acid and accumulation of the IAA anion (IAA/sup -/) in the alkaline cytoplasm; a saturable symport of IAA/sup -/ with one or more protons; a carrier-mediated efflux of IAA/sup -/ down a considerable electrochemical gradient. The efflux is greater from the basal than the apical end of cells and is thought to be responsible for the overall polarity of the process. This step is also the site of action of napthylphthalamic acid (NPA) and herbicides that inhibit polar transport but stimulate net accumulation of auxin by tissues and cells. We are using membrane vesicles as a simplified system for studying the mechanisms involved in the transport and accumulation of auxin. In particular, we are interested in determining the involvement of the transmembrane pH (pH/sub o/ < pH/sub i/) and voltage gradients (K/sup +/ diffusion potential, (K/sup +/)/sub in/ > (K/sup +/)/sub out/) in IAA uptake. 19 refs., 6 figs.

  5. Computational modelling of amino acid exchange and facilitated transport in placental membrane vesicles.

    PubMed

    Panitchob, N; Widdows, K L; Crocker, I P; Hanson, M A; Johnstone, E D; Please, C P; Sibley, C P; Glazier, J D; Lewis, R M; Sengers, B G

    2015-01-21

    Placental amino acid transport is required for fetal development and impaired transport has been associated with poor fetal growth. It is well known that placental amino acid transport is mediated by a broad array of specific membrane transporters with overlapping substrate specificity. However, it is not fully understood how these transporters function, both individually and as an integrated system. We propose that mathematical modelling could help in further elucidating the underlying mechanisms of how these transporters mediate placental amino acid transport. The aim of this work is to model the sodium independent transport of serine, which has been assumed to follow an obligatory exchange mechanism. However, previous amino acid uptake experiments in human placental microvillous plasma membrane vesicles have persistently produced results that are seemingly incompatible with such a mechanism; i.e. transport has been observed under zero-trans conditions, in the absence of internal substrates inside the vesicles to drive exchange. This observation raises two alternative hypotheses; (i) either exchange is not fully obligatory, or (ii) exchange is indeed obligatory, but an unforeseen initial concentration of amino acid substrate is present within the vesicle which could drive exchange. To investigate these possibilities, a mathematical model for tracer uptake was developed based on carrier mediated transport, which can represent either facilitated diffusion or obligatory exchange (also referred to as uniport and antiport mechanisms, respectively). In vitro measurements of serine uptake by placental microvillous membrane vesicles were carried out and the model applied to interpret the results based on the measured apparent Michaelis-Menten parameters Km and Vmax. In addition, based on model predictions, a new time series experiment was implemented to distinguish the hypothesised transporter mechanisms. Analysis of the results indicated the presence of a facilitated

  6. Reduction of dehydroascorbic acid at low pH.

    PubMed

    Wechtersbach, Luka; Cigić, Blaz

    2007-08-01

    Ascorbic acid and dehydroascorbic acid are unstable in aqueous solution in the presence of copper and iron ions, causing problems in the routine analysis of vitamin C. Their stability can be improved by lowering the pH below 2, preferably with metaphosphoric acid. Dehydroascorbic acid, an oxidised form of vitamin C, gives a relatively low response on the majority of chromatographic detectors, and is therefore routinely determined as the increase of ascorbic acid formed after reduction. The reduction step is routinely performed at a pH that is suboptimal for the stability of both forms. In this paper, the reduction of dehydroascorbic acid with tris-[2-carboxyethyl] phosphine (TCEP) at pH below 2 is evaluated. Dehydroascorbic acid is fully reduced with TCEP in metaphosphoric acid in less than 20 min, and yields of ascorbic acid are the same as at higher pH. TCEP and ascorbic acid formed by reduction, are more stable in metaphosphoric acid than in acetate or citrate buffers at pH 5, in the presence of redox active copper ions. The simple experimental procedure and low probability of artefacts are major benefits of this method, over those currently applied in a routine assay of vitamin C, performed on large number of samples.

  7. Lung lamellar bodies maintain an acidic internal pH

    SciTech Connect

    Chander, A.; Johnson, R.G.; Reicherter, J.; Fisher, A.B.

    1986-05-05

    The internal pH of lung lamellar bodies was investigated with membrane permeable basic amines. Isolated granular pneumocytes and isolated lung lamellar bodies exhibited fluorescence when exposed to 8 microM quinacrine, suggesting accumulation of this dye due to an acidic internal pH. Uptake of (/sup 14/C)methylamine by isolated lung lamellar bodies was measured to quantitate the intralamellar body pH. In KCl-ATP (10 mM) medium (pH 7.0), the accumulation ratio of methylamine (inside/outside) during 2-min incubation was 8.1 +/- 0.47 (mean +/- S.E., n = 8) indicating an internal pH of 6.1. Lamellar bodies accumulated methylamine almost 30-fold in K+-free mannitol medium indicating an internal pH of 5.6. The pH gradient across the lamellar body membrane decreased when external pH was decreased or when ATP was omitted. The pH gradient was also decreased by the addition of 10 mM NH/sub 4/Cl, 2 micrograms/ml nigericin, 0.02 mM N,N'-dicyclohexylcarbodiimide, or 1 mM N'-ethylmaleimide. These observations indicate that lamellar bodies maintain an acidic interior (pH 6.1 or below) which is generated by an energy-dependent process.

  8. High sequence homology between protein tyrosine acid phosphatase from boar seminal vesicles and human prostatic acid phosphatase.

    PubMed

    Wysocki, Paweł; Płucienniczak, Grazyna; Strzezek, Jerzy

    2009-01-01

    Boar seminal vesicle protein tyrosine acid phosphatase (PTAP) and human prostatic acid phosphatase (PAP) show high affinity for protein phosphotyrosine residues. The physico-chemical and kinetic properties of the boar and human enzymes are different. The main objective of this study was to establish the nucleotide sequence of cDNA encoding boar PTAP and compare it with that of human PAP cDNA. Also, the amino-acid sequence of boar PTAP was compared with the sequence of human PAP. PTAP was isolated from boar seminal vesicle fluid and sequenced. cDNA to boar seminal vesicle RNA was synthesized, amplified by PCR, cloned in E. coli and sequenced. The obtained N-terminal amino-acid sequence of boar PTAP showed 92% identity with the N-terminal amino-acid sequence of human PAP. The determined sequence of a 354 bp nucleotide fragment (GenBank accession number: GQ184596) showed 90% identity with the corresponding sequence of human PAP. On the basis of this sequence a 118 amino acid fragment of boar PTAP was predicted. This fragment showed 89% identity with the corresponding fragment of human PAP and had a similar hydropathy profile. The compared sequences differ in terms of their isoelectric points and amino-acid composition. This may explain the differences in substrate specificity and inhibitor resistance of boar PTAP and human PAP.

  9. Functional reconstitution of the. gamma. -aminobutyric acid transporter from synaptic vesicles using artificial ion gradients

    SciTech Connect

    Hell, J.W.; Edelmann, L.; Hartinger, J.; Jahn, R. )

    1991-12-24

    The {gamma}-aminobutyric acid transporter of rat brain synaptic vesicles was reconstituted in proteoliposomes, and its activity was studied in response to artificially created membrane potentials or proton gradients. Changes of the membrane potential were monitored using the dyes oxonol VI and 3,3{prime}-diisopropylthiodicarbocyanine iodide, and changes of the H{sup +} gradient were followed using acridine orange. An inside positive membrane potential was generated by the creation of an inwardly directed K{sup +} gradient and the subsequent addition of valinomycin. Under these conditions, valinomycin evoked uptake of ({sup 3}H)GABA which was saturable. Similarly, ({sup 3}H)glutamate uptake was stimulated by valinomycin, indicating that both transporters can be driven by the membrane potential. Proton gradients were generated by the incubation of K{sup +}-loaded proteoliposomes in a buffer free of K{sup +} or Na{sup +} ions and the subsequent addition of nigericin. Proton gradients were also generated via the endogenous H{sup +} ATPase by incubation of K{sup +}-loaded proteoliposomes in equimolar K{sup +} buffer in the presence of valinomycin. These proton gradients evoked nonspecific, nonsaturable uptake of GABA and {beta}-alanine but not of glycine in proteoliposomes as well as protein-free liposomes. Therefore, transporter activity was monitored using glycine as an alternative substrate. Proton gradients generated by both methods elicited saturable glycine uptake in proteoliposomes. Together, these data confirm that the vesicular GABA transporter can be energized by both the membrane potential and the pH gradient and show that transport can be achieved by artificial gradients independently of the endogenous proton ATPase.

  10. Factors influencing uptake and retention of amino-containing drugs in large unilamellar vesicles exhibiting transmembrane pH gradients.

    PubMed

    Maurer-Spurej, E; Wong, K F; Maurer, N; Fenske, D B; Cullis, P R

    1999-01-12

    The level of uptake and retention of amino-containing drugs in large unilamellar vesicles (LUVs) following uptake in response to a transmembrane pH gradient (DeltapH) can vary dramatically depending on the drug. For example, the anticancer drugs doxorubicin and epirubicin can be readily retained, whereas the anticancer drug vincristine and the antibiotic ciprofloxacin tend to leak out rapidly. In this investigation, we examine the influence of the hydrophobicity of the entrapped amines (that induce the DeltapH) and the anionic lipid content of the LUV on drug retention. It is shown that entrapment of increasingly hydrophobic monoamines (methylamine to amylamine) all lead to an induced DeltapH of 3 units and essentially complete drug uptake under the conditions employed, but do not lead to improved retention of vincristine and ciprofloxacin. However, significantly improved retention could be achieved by substitution of the anionic lipid distearoylphosphatidylglycerol (DSPG) for distearoylphosphatidylcholine (DSPC) in the LUV bilayer. Further, it is shown that if the induced DeltapH is reduced to 1.4 units (driven by entrapped diamine) nearly 100% accumulation of doxorubicin and epirubicin could be achieved, whereas only 25% loading for vincristine and ciprofloxacin was observed. Taken together these results provide methodology for improving (weak base) drug retention in liposomes and indicate that drugs that can partition into the lipid bilayer exhibit improved uptake and retention characteristics.

  11. Chain-length heterogeneity allows for the assembly of fatty acid vesicles in dilute solutions.

    PubMed

    Budin, Itay; Prwyes, Noam; Zhang, Na; Szostak, Jack W

    2014-10-07

    A requirement for concentrated and chemically homogeneous pools of molecular building blocks would severely restrict plausible scenarios for the origin of life. In the case of membrane self-assembly, models of prebiotic lipid synthesis yield primarily short, single-chain amphiphiles that can form bilayer vesicles only at very high concentrations. These high critical aggregation concentrations (cacs) pose significant obstacles for the self-assembly of single-chain lipid membranes. Here, we examine membrane self-assembly in mixtures of fatty acids with varying chain lengths, an expected feature of any abiotic lipid synthesis. We derive theoretical predictions for the cac of mixtures by adapting thermodynamic models developed for the analogous phenomenon of mixed micelle self-assembly. We then use several complementary methods to characterize aggregation experimentally, and find cac values in close agreement with our theoretical predictions. These measurements establish that the cac of fatty acid mixtures is dramatically lowered by minor fractions of long-chain species, thereby providing a plausible route for protocell membrane assembly. Using an NMR-based approach to monitor aggregation of isotopically labeled samples, we demonstrate the incorporation of individual components into mixed vesicles. These experiments suggest that vesicles assembled in dilute, mixed solutions are depleted of the shorter-chain-length lipid species, a finding that carries implications for the composition of primitive cell membranes.

  12. Photoreactions of bacteriorhodopsin at acid pH

    SciTech Connect

    Varo, G.; Lanyi, J.K. )

    1989-12-01

    It has been known that bacteriorhodopsin, the retinal protein in purple membrane which functions as a light-driven proton pump, undergoes reversible spectroscopic changes at acid pH. The absorption spectra of various bacteriorhodopsin species were estimated from measured spectra of the mixtures that form at low pH, in the presence of sulfate and chloride. The dependency of these on pH and the concentration of Cl- fit a model in which progressive protonation of purple membrane produces blue membrane, which will bind, with increasing affinity as the pH is lowered, chloride ions to produce acid purple membrane. Transient spectroscopy with a multichannel analyzer identified the intermediates of the photocycles of these altered pigments, and described their kinetics. Blue membrane produced red-shifted KL-like and L-like products, but no other photointermediates, consistent with earlier suggestions. Unlike others, however, we found that acid purple membrane exhibited a very different photocycle: its first detected intermediate was not like KL in that it was much more red-shifted, and the only other intermediate detectable resembled the O species of the bacteriorhodopsin photocycle. An M-like intermediate, with a deprotonated Schiff base, was not found in either of these photocycles. There are remarkable similarities between the photoreactions of the acid forms of bacteriorhodopsin and the chloride transport system halorhodopsin, where the Schiff base deprotonation seems to be prevented by lack of suitable aspartate residues, rather than by low pH.

  13. Gas vesicles.

    PubMed Central

    Walsby, A E

    1994-01-01

    The gas vesicle is a hollow structure made of protein. It usually has the form of a cylindrical tube closed by conical end caps. Gas vesicles occur in five phyla of the Bacteria and two groups of the Archaea, but they are mostly restricted to planktonic microorganisms, in which they provide buoyancy. By regulating their relative gas vesicle content aquatic microbes are able to perform vertical migrations. In slowly growing organisms such movements are made more efficiently than by swimming with flagella. The gas vesicle is impermeable to liquid water, but it is highly permeable to gases and is normally filled with air. It is a rigid structure of low compressibility, but it collapses flat under a certain critical pressure and buoyancy is then lost. Gas vesicles in different organisms vary in width, from 45 to > 200 nm; in accordance with engineering principles the narrower ones are stronger (have higher critical pressures) than wide ones, but they contain less gas space per wall volume and are therefore less efficient at providing buoyancy. A survey of gas-vacuolate cyanobacteria reveals that there has been natural selection for gas vesicles of the maximum width permitted by the pressure encountered in the natural environment, which is mainly determined by cell turgor pressure and water depth. Gas vesicle width is genetically determined, perhaps through the amino acid sequence of one of the constituent proteins. Up to 14 genes have been implicated in gas vesicle production, but so far the products of only two have been shown to be present in the gas vesicle: GvpA makes the ribs that form the structure, and GvpC binds to the outside of the ribs and stiffens the structure against collapse. The evolution of the gas vesicle is discussed in relation to the homologies of these proteins. Images PMID:8177173

  14. Activation of protein kinase C by lysophosphatidic acid: dependence on composition of phospholipid vesicles.

    PubMed Central

    Sando, J J; Chertihin, O I

    1996-01-01

    Lysophosphatidic acid (LPA) has attracted recent attention as a major serum-derived regulator implicated in responses to vascular injury and inflammation, in tumour invasiveness and in neuronal signalling and remodelling. Although the possibility of a specific G-protein-coupled LPA receptor protein has been suggested, characterization of such a receptor is lacking. Since LPA can activate protein kinase C (PKC) pathways in many cells and PKC activators mimic many LPA effects, the possibility of more direct LPA effects on PKC was investigated. Phosphatidylcholine (PC)/phosphatidylserine (PS)/diacylglycerol (DAG) lipid vesicles of defined acyl chain composition were used to activate the enzyme. At total concentrations of saturated PC/PS + DAG vesicles (2-3 mM) that provided maximal PKC activation, 1-10 mol % [18:1]-LPA led to a further approx. 2-fold activation of PKC alpha. At lower lipid concentrations, a greater increase was observed with LPA concentrations up to 16-20 mol %. Higher concentrations of LPA were inhibitory. The LPA activation of PKC was dependent on the presence of DAG, PS and Ca2+. [18:1]-Lysophosphatidylcholine produced similar PKC activation in PC/PS/DAG vesicles. [14:0]-LPA was less effective, and longer-chain saturated lysolipids were ineffective. In unsaturated PC/PS vesicles, very little to no effect of LPA was discernable. These results suggest that physiologically or pathologically relevant concentrations of LPA can contribute to PKC activation depending on the composition of the lipid membrane. We hypothesize that LPA may affect the formation of lipid domains that are recognized by the enzyme. PMID:8713089

  15. Organic acids make Escherichia coli more resistant to pulsed electric fields at acid pH.

    PubMed

    Somolinos, M; García, D; Mañas, P; Condón, S; Pagán, R

    2010-01-01

    Stationary growth phase cells of Escherichiacoli were more pulsed electric fields (PEF) resistant in citrate-phosphate McIlvaine buffer at pH 4.0 than at pH 7.0. The greater PEF resistance was also confirmed in fruit juices of similar acid pH. In this work we studied whether the higher PEF resistance of E. coli at acid pH was due to the low pH itself or to the interaction of the components of the treatment medium with the cells. The protective effect on E. coli cells was due to the presence of organic acids such as citric, acetic, lactic or malic at pH 4.0. The protective effect of citric acid at pH 4.0 depended on its concentration. A linear relationship was observed between the Log(10) of the citric acid concentration and the degree of inactivation. Organic acids contained in laboratory treatment media (citrate-phosphate buffer) or in fruit juices did not sensitize E. coli cells to PEF but, on the contrary, they induced a protective effect that made E. coli cells more resistant at pH 4.0 than at neutral pH. This work could be useful for improving food preservation by PEF technology and it contributes to the knowledge of the mechanism of microbial inactivation by PEF.

  16. Clavulanic acid increases dopamine release in neuronal cells through a mechanism involving enhanced vesicle trafficking.

    PubMed

    Kost, Gina Chun; Selvaraj, Senthil; Lee, Young Bok; Kim, Deog Joong; Ahn, Chang-Ho; Singh, Brij B

    2011-10-24

    Clavulanic acid is a CNS-modulating compound with exceptional blood-brain barrier permeability and safety profile. Clavulanic acid has been proposed to have anti-depressant activity and is currently entering Phase IIb clinical trials for the treatment of Major Depressive Disorder (MDD). Studies have also shown that clavulanic acid suppresses anxiety and enhances sexual functions in rodent and primate models by a mechanism involving central nervous system (CNS) modulation, although its detailed mechanism of action has yet to be elucidated. To further examine its potential as a CNS modulating agent as well as its mechanism of action, we investigated the effects of clavulanic acid in neuronal cells. Our results indicate that clavulanic acid enhances dopamine release in PC12 and SH-SY5Y cells without affecting dopamine synthesis. Furthermore, using affinity chromatography we were able to identify two proteins, Munc18-1 and Rab4 that potentially bind to clavulanic acid and play a critical role in neurosecretion and the vesicle trafficking process. Consistent with this result, an increase in the translocation of Munc18-1 and Rab4 from the cytoplasm to the plasma membrane was observed in clavulanic acid treated cells. Overall, these data suggest that clavulanic acid enhances dopamine release in a mechanism involving Munc18-1 and Rab4 modulation and warrants further investigation of its therapeutic use in CNS disorders, such as depression. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  17. Spontaneous vesicle formation from sodium salt of acidic sophorolipid and its application as a skin penetration enhancer.

    PubMed

    Imura, Tomohiro; Morita, Tomotake; Fukuoka, Tokuma; Ryu, Mizuyuki; Igarashi, Keisuke; Hirata, Yoshihiko; Kitamoto, Dai

    2014-01-01

    In this study, spontaneous vesicle formation from the sodium salt of acidic sophorolipid (SLNa) was observed, and its potential application as a skin penetration enhancer for triterpene glycosides extracted from the fruits of Siraitia grosvenorii Swingle was then investigated. Dynamic light scattering (DLS) measurements of the SLNa assemblies prepared by the gentle mixing of SLNa with water (1%) showed their hydrodynamic radius (Rh) to be 96.2 nm, and their structure was assigned to be vesicles by freeze-fracture electron microscopy (FF-TEM). DLS and FF-TEM also revealed that the size of the vesicles increased with an increase in the concentration of the triterpene glycosides, indicating that the triterpene glycosides were incorporated into the SLNa vesicles. The results of an in vitro skin permeation assay, after loading the SLNa vesicles on a 3D cultured skin model, showed that the amount of SLNa that penetrated though the skin model increased with time. It was also found that the amount of permeated mogroside, which is the main active component of triterpene glycosides, was significantly enhanced by the SLNa vesicle formulation. These results clearly demonstrated that spontaneously formed vesicles composed of the bolaamphiphile SLNa are useful for application as penetration enhancers for active ingredients such as mogroside V.

  18. Acidic pH environment induces autophagy in osteoblasts

    PubMed Central

    Zhang, Zhichao; Lai, Qingguo; Li, Yanan; Xu, Chao; Tang, Xiaopeng; Ci, Jiangbo; Sun, Shaolong; Xu, Bingbing; Li, Yan

    2017-01-01

    Osteoblasts (OBs) play an important role in bone fracture healing, yet the extreme adverse microenvironment in fracture sites has a negative impact on the survival of OBs. Therefore, it is important to study how OBs behave in the complex fracture microenvironment. Studies have shown that autophagy plays a pivotal role in maintaining cellular homeostasis and defending the cell against adverse microenvironments. In this study we found the induction of autophagy in OBs at femoral bone fracture sites, which may be a result of ischemia, oxidative stress and hypoxia within the local area. At fracture sites a low pH environment also developed. Until now it has been unclear whether the induction of autophagy in osteoblasts is triggered by the acidic pH environment. Therefore, we cultured OBs in vitro in media of different pH values, and found both autophagy and apoptosis increased in OBs in acidic conditions. However, when autophagy inhibitor chloroquine (CQ) was used, apoptosis increased significantly compared with that without CQ. Thus indicating that inhibition of autophagy may promote apoptosis in OBs in an acidic environment, which may provide a new therapeutic strategy to decrease cell apoptosis in OBs through the use of drugs that modulate the autophagic state. PMID:28382973

  19. Measurement of luminal pH of acidic stores as a readout for NAADP action.

    PubMed

    Galione, Antony; Chuang, Kai-Ting; Funnell, Tim M; Davis, Lianne C; Morgan, Anthony J; Ruas, Margarida; Parrington, John; Churchill, Grant C

    2014-10-01

    In addition to mobilizing Ca²⁺, NAADP plays a role in modulating the luminal pH (pHL) of acidic stores of the endolysosomal system. The effects of NAADP on pHL have been most extensively studied in the sea urchin egg, both in the intact egg and in egg homogenates. Related observations have also been made in mammalian systems (e.g., guinea pig atrial myocytes and pancreatic acinar cells). Although the connection between Ca²⁺ mobilization and increase in pHL is not understood, pHL can be a useful parameter to measure when studying NAADP-mediated signaling. This protocol describes the fluorescent measurement of pHL of acidic stores. It relies on the use of acridine orange (AO), a standard dye for pHL. AO selectively accumulates to high concentrations in the lumen of organelles as a function of acidity; at these high concentrations it self-quenches. When pHL increases, some AO is lost from the vesicle. As a result, the lower luminal AO concentration relieves the quenching and fluorescence increases in the lumen.

  20. Roles of alkaline phosphatase and labile internal mineral in matrix vesicle-mediated calcification. Effect of selective release of membrane-bound alkaline phosphatase and treatment with isosmotic pH 6 buffer.

    PubMed

    Register, T C; McLean, F M; Low, M G; Wuthier, R E

    1986-07-15

    The roles of alkaline phosphatase and labile internal mineral in matrix vesicle-mediated mineralization have been studied by selectively releasing the enzyme from a wide variety of matrix vesicle preparations using treatment with a bacterial phosphatidylinositol-specific phospholipase C and by demineralization of the vesicles using isosmotic pH 6 buffer. Following depletion of 50-90% of the alkaline phosphatase activity or treatment with citrate buffer, the vesicles were tested for their ability to accumulate 45Ca2+ and 32Pi from a synthetic cartilage lymph. Removal of alkaline phosphatase by phospholipase C treatment caused two principal effects, depending on the matrix vesicle preparation. In rapidly mineralizing vesicle fractions which did not require organic phosphate esters (Po) to accumulate mineral ions, release of alkaline phosphatase had only a minor effect. In slowly mineralizing vesicles preparations or those dependent on Po substrates for mineral ion uptake, release of alkaline phosphatase caused significant loss of mineralizing activity. The activity of rapidly calcifying vesicles was shown to be dependent on the presence of labile internal mineral, as demonstrated by major loss in activity when the vesicles were decalcified by various treatments. Ion uptake by demineralized vesicles or those fractionated on sucrose step gradients required Po and was significantly decreased by alkaline phosphatase depletion. Uptake of Pi, however, was not coupled with hydrolysis of the Po substrate. These findings argue against a direct role for alkaline phosphatase as a porter in matrix vesicle Pi uptake, contrary to previous postulates. The results emphasize the importance of internal labile mineral in rapid uptake of mineral ions by matrix vesicles.

  1. Active transport of. gamma. -aminobutyric acid and glycine into synaptic vesicles

    SciTech Connect

    Kish, P.E.; Fischer-Bovenkerk, C.; Ueda, T. )

    1989-05-01

    Although {gamma}-aminobutyric acid (GABA) and glycine are recognized as major amino acid inhibitory neurotransmitters in the central nervous system, their storage is poorly understood. In this study the authors have characterized vesicular GABA and glycine uptakes in the cerebrum and spinal cord, respectively. They present evidence that GABA and glycine are each taken up into isolated synaptic vesicles in an ATP-dependent manner and that the uptake is driven by an electrochemical proton gradient. Uptake for both amino acids exhibited kinetics with low affinity similar to a vesicular glutamate uptake. The ATP-dependent GABA uptake was not inhibited by the putative amino acid neurotransmitters glycine, taurine, glutamate, or aspartate or by GABA analogs, agonists, and antagonists. Similarly, ATP-dependent glycine uptake was hardly affected by GABA, taurine, glutamate, or aspartate or by glycine analogs or antagonists. The GABA uptake was not affected by chloride, which is in contrast to the uptake of the excitatory neurotransmitter glutamate, whereas the glycine uptake was slightly stimulated by low concentrations of chloride. Tissue distribution studies indicate that the vesicular uptake systems for GABA, glycine, and glutamate are distributed in different proportions in the cerebrum and spinal cord. These results suggest that the vesicular uptake systems for GABA, glycine, and glutamate are distinct from each other.

  2. The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells.

    PubMed

    Glomski, Ian J; Gedde, Margaret M; Tsang, Albert W; Swanson, Joel A; Portnoy, Daniel A

    2002-03-18

    Listeria monocytogenes is a facultative intracellular bacterial pathogen that escapes from a phagosome and grows in the host cell cytosol. The pore-forming cholesterol-dependent cytolysin, listeriolysin O (LLO), mediates bacterial escape from vesicles and is approximately 10-fold more active at an acidic than neutral pH. By swapping dissimilar residues from a pH-insensitive orthologue, perfringolysin O (PFO), we identified leucine 461 as unique to pathogenic Listeria and responsible for the acidic pH optimum of LLO. Conversion of leucine 461 to the threonine present in PFO increased the hemolytic activity of LLO almost 10-fold at a neutral pH. L. monocytogenes synthesizing LLO L461T, expressed from its endogenous site on the bacterial chromosome, resulted in a 100-fold virulence defect in the mouse listeriosis model. These bacteria escaped from acidic phagosomes and initially grew normally in cells and spread cell to cell, but prematurely permeabilized the host membrane and killed the cell. These data show that the acidic pH optimum of LLO results from an adaptive mutation that acts to limit cytolytic activity to acidic vesicles and prevent damage in the host cytosol, a strategy also used by host cells to compartmentalize lysosomal hydrolases.

  3. Preferential packing of acidic glycosidases and proteases into Bacteroides outer membrane vesicles.

    PubMed

    Elhenawy, Wael; Debelyy, Mykhaylo O; Feldman, Mario F

    2014-03-11

    Outer membrane vesicles (OMV) are spherical membranous structures released from the outer membrane (OM) of Gram-negative bacteria. OMV have been proposed to play several different roles during both pathogenesis and symbiosis. Despite the fact that OMV were described several decades ago, their biogenesis is a poorly characterized process. Whether OMV are produced by an active mechanism or by passive disintegration of the OM is a still matter of controversy. Bacteroides fragilis and Bacteroides thetaiotaomicron are important members of the human microbiota. In this work, we determined and compared the protein compositions of OM and OMV from B. fragilis and B. thetaiotaomicron. SDS-PAGE analysis of both fractions revealed dramatically different protein profiles. Proteomic analysis of OM and OMV in B. fragilis identified more than 40 proteins found exclusively in OMV and more than 30 proteins detectable only in the OM. The OMV-specific proteome showed a high prevalence of glycosidases and proteases, some of which were shown to be active in vitro. Similar results were obtained for B. thetaiotaomicron. Most of the OMV-exclusive proteins were acidic. Based on these results, we propose that these species possess machinery devoted to selectively pack acidic proteins into the OMV. These OMV equipped with hydrolytic enzymes could help in securing nutrients for the benefit of the whole bacterial community present in the microbiota, uncovering a novel function for bacterial OMV. IMPORTANCE The members of genus Bacteroides are key players in the symbiosis between the human host and the gut microbiota. It is known for its ability to degrade a wide variety of glycans that are not substrates for human glycosidases. The cleaved glycans can be utilized by Bacteroides and other microbiota members, resulting in the production of short-chain fatty acids that are beneficial for the host. Although members of the genus Bacteroides are known to secrete different hydrolases, their secretion

  4. Inhibition by Levorphanol and Related Drugs of Amino Acid Transport by Isolated Membrane Vesicles from Escherichia coli

    PubMed Central

    Holland, Mary J. C.; Simon, Eric J.

    1975-01-01

    Levorphanol inhibits the transport of the amino acids proline and lysine by cytoplasmic membrane vesicles derived from Escherichia coli. The degree of inhibition increases with increasing levorphanol concentration and ranges from 26% at 10−6 M levorphanol to 92% at 10−3 M levorphanol. The effect is independent of the energy source, since levorphanol inhibits proline uptake to the same extent in the presence of 20 mM d-lactate or 20 mM succinate and in the absence of an exogenous energy source. Levorphanol does not irreversibly alter the ability of membrane vesicles to transport proline, since incubation of membrane vesicles for 15 min in the presence of 0.25 mM levorphanol, a concentration which inhibits proline transport by more than 75%, has no effect on the rate of proline transport by these vesicles once the drug is removed. Both the maximum velocity and the Km of proline transport are modified by levorphanol, hence, the type of inhibition produced by levorphanol is mixed. The inhibitor constant (Ki) for levorphanol inhibition of proline transport is approximately 3 × 10−4 M. Membrane vesicles incubated in the presence of levorphanol accumulate much less proline at the steady state than do control vesicles. Furthermore, the addition of levorphanol to membrane vesicles preloaded to the steady state with proline produces a marked net efflux of proline. Levorphanol does not block either temperature-induced efflux or exchange of external proline with [14C]proline present in the intravesicular pool. Dextrorphan, the enantiomorph of levorphanol, and levallorphan, the N-allyl analogue of levorphanol, inhibit proline and lysine transport in a similar manner. Possible mechanisms of the effects of these drugs on cell membranes are discussed. PMID:1096802

  5. Compartmentation of malic acid in mesophyll cells of Kalanchoee daigremontiana: indications of a intracellular cytosolic vesicle transport mechanism

    SciTech Connect

    Balsamo, R.A.; Uribe, E.G.

    1987-04-01

    Leaf tissue was harvested over a 24hr period at one to three hour intervals. The malic acid levels in the tissue were assayed spectrophotometrically and the percent cell volume occupied by cytosolic vesicles in replicate samples was determined. The total volume of the cytosolic vesicles fluctuated throughout the photoperiod concommitantly with malic acid concentrations present in the tissue. An intact leaf tissue section (10.2cm/sup 2/) was radiolabeled with /sup 14/CO/sub 2/ seven hours into the dark period for thirty minutes. Two dimensional thin layer chromatography and electrophoresis of the tissue determined that 96% of the label was incorporated into malic acid. A freeze substitution procedure was initiated followed by microautoradiography (Fisher 1971) which allowed for the tracing of intracellular malic acid migration and compartmentation within the mesophyll cells. The results and interpretation of this experiment will be presented.

  6. Dietary lipids modify the fatty acid composition of cartilage, isolated chondrocytes and matrix vesicles.

    PubMed

    Xu, H; Watkins, B A; Adkisson, H D

    1994-09-01

    The effects of dietary lipids on the fatty acid composition of hyaline cartilage, epiphyseal chondrocytes (EC) and matrix vesicles (MV) were evaluated in chicks. A basal semipurified diet was fed to chicks containing one of the following lipid sources at 70 g/kg: soybean oil, butter+corn oil, margarine+corn oil or menhaden oil+corn oil (MEC). Articular and epiphyseal growth cartilage were isolated from the proximal tibiotarsus; EC and MV were subsequently released by trypsin (EC 3.4.21.4) and collagenase (EC 3.4.24.3) digestion followed by ultracentrifugation. The fatty acid composition of polar lipids in chick epiphyseal cartilage at three and six weeks, as well as articular cartilage, EC and MV at eight weeks of age revealed the presence of high levels of saturated and monounsaturated fatty acids (up to 85.5%) but low levels of n-6 polyunsaturated fatty acids (PUFA) (2.6-10.2%). Mead acid (20:3n-9, > 3%) was also present in cartilage, EC and MV lipids, and was unaffected by the dietary lipid treatments. Total n-3 PUFA concentrations were the highest in cartilage, EC and MV of chicks consuming MEC. Feeding MEC lowered the levels of 20:4n-6 in cartilage, but increased 20:5n-3 levels. The data are consistent with those reported previously which showed that cartilage tissues are low in n-6 PUFA and that they contain 20:3n-9. We furthermore demonstrated that the PUFA composition of cartilage can be modified by dietary lipids.

  7. Effect of dietary fatty acids on jejunal and ileal oleic acid uptake by rat brush border membrane vesicles.

    PubMed

    Prieto, R M; Stremmel, W; Sales, C; Tur, J A

    1996-04-18

    To test the effect of dietary fatty acids on fatty acid uptake, the influx kinetics of a representative long-chain fatty acid, 3H-oleic acid, in both the jejunum and ileum of rats has been studied using brush border membrane vesicles (BBMV). Animals were fed with semipurified diets containing 5 g fat/100 g diet, as corn oil (control group), safflower oil (unsaturated group) and coconut oil hydrogenated (saturated group). With increasing unbound oleate concentration in the medium, the three dietary groups showed saturable kinetics in both jejunal and ileal BBMV (controls: Vmax = 0.15 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 136 +/- 29.1 nmol for jejunum, and Vmax = 0.23 +/- 0.03 nmol x mg protein-1 x 5 min-1 and Km = 196 +/- 50.3 nmol for ileum; unsaturated: Vmax = 0.28 +/- 0.05 nmol x mg protein-1 x 5 min-1 and Km = 242.7 +/- 91.8 nmol for jejunum, and Vmax = 1.29 +/- 0.06 nmol x mg protein-1 x 5 min-1 and Km = 509.8 +/- 97.5 nmol for ileum; saturated: Vmax = 0.03 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 124.5 +/- 72.6 nmol for jejunum, and Vmax = 0.04 +/- 0.01 nmol x mg protein -1.5 min-1 and Km = 205.6 +/- 85.3 nmol for ileum). These results support the theory that feeding an isocaloric diet containing only unsaturated fatty acids enhanced oleic acid uptake, and feeding an isocaloric diet containing only saturated fatty acids decreased oleic acid uptake. The results obtained in the present work also show the adaptative ability of jejunum and ileum to the type of dietary fat.

  8. Cholesterol supports the retinoic acid-induced synaptic vesicle formation in differentiating human SH-SY5Y neuroblastoma cells.

    PubMed

    Sarkanen, Jertta-Riina; Nykky, Jonna; Siikanen, Jutta; Selinummi, Jyrki; Ylikomi, Timo; Jalonen, Tuula O

    2007-09-01

    Synaptic vesicle formation, vesicle activation and exo/endocytosis in the pre-synaptic area are central steps in neuronal communication. The formation and localization of synaptic vesicles in human SH-SY5Y neuroblastoma cells, differentiated with 12-o-tetradecanoyl-phorbol-13-acetate, dibutyryl cyclic AMP, all-trans-retinoic acid (RA) and cholesterol, was studied by fluorescence microscopy and immunocytochemical methods. RA alone or together with cholesterol, produced significant neurite extension and formation of cell-to-cell contacts. Synaptic vesicle formation was followed by anti-synaptophysin (SypI) and AM1-43 staining. SypI was only weakly detected, mainly in cell somata, before 7 days in vitro, after which it was found in neurites. Depolarization of the differentiated cells with high potassium solution increased the number of fluorescent puncta, as well as SypI and AM1-43 co-localization. In addition to increase in the number of synaptic vesicles, RA and cholesterol also increased the number and distribution of lysosome-associated membrane protein 2 labeled lysosomes. RA-induced Golgi apparatus fragmentation was partly avoided by co-treatment with cholesterol. The SH-SY5Y neuroblastoma cell line, differentiated by RA and cholesterol and with good viability in culture, is a valuable tool for basic studies of neuronal metabolism, specifically as a model for dopaminergic neurons.

  9. A Historical and Evolutionary Perspective on Circulating Nucleic Acids and Extracellular Vesicles: Circulating Nucleic Acids as Homeostatic Genetic Entities.

    PubMed

    Aucamp, Janine; Bronkhorst, Abel Jacobus; Pretorius, Piet J

    2016-01-01

    The quantitative and qualitative differences of circulating nucleic acids (cirNAs) between healthy and diseased individuals have motivated researchers to utilize these differences in the diagnosis and prognosis of various pathologies. The position maintained here is that reviewing the rather neglected early work associated with cirNAs and extracellular vesicles (EVs) is required to fully describe the nature of cirNAs. This review consists of an empirically up-to-date schematic summary of the major events that developed and integrated the concepts of heredity, genetic information and cirNAs. This reveals a clear pattern implicating cirNA as a homeostatic entity or messenger of genetic information. The schematic summary paints a picture of how cirNAs may serve as homeostatic genetic entities that promote synchrony of both adaptation and damage in tissues and organs depending on the source of the message.

  10. Preferential Packing of Acidic Glycosidases and Proteases into Bacteroides Outer Membrane Vesicles

    PubMed Central

    Elhenawy, Wael; Debelyy, Mykhaylo O.; Feldman, Mario F.

    2014-01-01

    ABSTRACT Outer membrane vesicles (OMV) are spherical membranous structures released from the outer membrane (OM) of Gram-negative bacteria. OMV have been proposed to play several different roles during both pathogenesis and symbiosis. Despite the fact that OMV were described several decades ago, their biogenesis is a poorly characterized process. Whether OMV are produced by an active mechanism or by passive disintegration of the OM is a still matter of controversy. Bacteroides fragilis and Bacteroides thetaiotaomicron are important members of the human microbiota. In this work, we determined and compared the protein compositions of OM and OMV from B. fragilis and B. thetaiotaomicron. SDS-PAGE analysis of both fractions revealed dramatically different protein profiles. Proteomic analysis of OM and OMV in B. fragilis identified more than 40 proteins found exclusively in OMV and more than 30 proteins detectable only in the OM. The OMV-specific proteome showed a high prevalence of glycosidases and proteases, some of which were shown to be active in vitro. Similar results were obtained for B. thetaiotaomicron. Most of the OMV-exclusive proteins were acidic. Based on these results, we propose that these species possess machinery devoted to selectively pack acidic proteins into the OMV. These OMV equipped with hydrolytic enzymes could help in securing nutrients for the benefit of the whole bacterial community present in the microbiota, uncovering a novel function for bacterial OMV. PMID:24618254

  11. Encapsulation of Nucleic Acids into Giant Unilamellar Vesicles by Freeze-Thaw: a Way Protocells May Form

    NASA Astrophysics Data System (ADS)

    Qiao, Hai; Hu, Na; Bai, Jin; Ren, Lili; Liu, Qing; Fang, Liaoqiong; Wang, Zhibiao

    2016-11-01

    Protocells are believed to consist of a lipid membrane and encapsulated nucleic acid. As the lipid membrane is impermeable to macromolecules like nucleic acids, the processes by which nucleic acids become encapsulated inside lipid membrane compartments are still unknown. In this paper, a freeze-thaw method was modified and applied to giant unilamellar vesicles (GUVs) and deoxyribonucleic acid (DNA) in mixed solution resulting in the efficient encapsulation of 6.4 kb plasmid DNA and similar length linear DNA into GUVs. The mechanism of encapsulation was followed by observing the effect of freeze-thaw temperatures on GUV morphological change, DNA encapsulation and ice crystal formation, and analyzing their correlation. Following ice crystal formation, the shape of spherical GUVs was altered and membrane integrity was damaged and this was found to be a necessary condition for encapsulation. Heating alone had no effects on DNA encapsulation, but was helpful for restoring the spherical shape and membrane integrity of GUVs damaged during freezing. These results suggested that freeze-thaw could promote the encapsulation of DNA into GUVs by a mechanism: the vesicle membrane was breached by ice crystal formation during freezing, DNA entered into damaged GUVs through these membrane gaps and was encapsulated after the membrane was resealed during the thawing process. The process described herein therefore describes a simple way for the encapsulation of nucleic acids and potentially other macromolecules into lipid vesicles, a process by which early protocells might have formed.

  12. Permeability and Channel-Mediated Transport of Boric Acid across Membrane Vesicles Isolated from Squash Roots1

    PubMed Central

    Dordas, Christos; Chrispeels, Maarten J.; Brown, Patrick H.

    2000-01-01

    Boron is an essential micronutrient for plant growth and the boron content of plants differs greatly, but the mechanism(s) of its uptake into cells is not known. Boron is present in the soil solution as boric acid and it is in this form that it enters the roots. We determined the boron permeability coefficient of purified plasma membrane vesicles obtained from squash (Cucurbita pepo) roots and found it to be 3 × 10−7 ±1.4 × 10−8 cm s−1, six times higher than the permeability of microsomal vesicles. Boric acid permeation of the plasma membrane vesicles was partially inhibited (30%–39%) by mercuric chloride and phloretin, a non-specific channel blocker. The inhibition by mercuric chloride was readily reversible by 2-mercaptoethanol. The energy of activation for boron transport into the plasma membrane vesicles was 10.2 kcal mol−1. Together these data indicate that boron enters plant cells in part by passive diffusion through the lipid bilayer of the plasma membrane and in part through proteinaceous channels. Expression of the major intrinsic protein (MIP) PIP1 in Xenopus laevis oocytes resulted in a 30% increase in the boron permeability of the oocytes. Other MIPs tested (PIP3, MLM1, and GlpF) did not have this effect. We postulate that certain MIPs, like those that have recently been shown to transport small neutral solutes, may also be the channels through which boron enters plant cells. PMID:11080310

  13. A rocket-like encapsulation and delivery system with two-stage booster layers: pH-responsive poly(methacrylic acid)/poly(ethylene glycol) complex-coated hollow silica vesicles.

    PubMed

    Lay, Chee Leng; Kumar, Jatin N; Liu, Connie K; Lu, Xuehong; Liu, Ye

    2013-10-01

    Rocket-like vesicles formed are composed of poly(acrylic aicd) (PMAA )/poly(ethylene glycol) (PEG) complex coated hollow silica spheres, and the structure and composition of the vesicles are characterized using TGA, (1)H NMR, FTIR, and TEM. Although only one-third of EG units of PEG brushes grafted to hollow silica spheres form the complex with PMAA via hydrogen bonding, the first "booster" layer composed of PMAA/PEG complex can provide secure encapsulation of model compound calcein blue under an acidic condition. The second "booster" layer composed of PEG brushes can be formed by changing acidic pH to 7.4 through the disassociation of the PMAA/PEG complex. A higher molecular weight PMAA exhibits a faster disassembly due to the formation of a looser PMAA/PEG complex on the surfaces of hollow silica spheres.

  14. Cytoplasmic pH mediates pH taxis and weak-acid repellent taxis of bacteria.

    PubMed

    Kihara, M; Macnab, R M

    1981-03-01

    Bacteria migrate away from an acid pH and from a number of chemicals, including organic acids such as acetate; the basis for detection of these environmental cues has not been demonstrated. Membrane-permeant weak acids caused prolonged tumbling when added to Salmonella sp. or Escherichia coli cells at pH 5.5. Tethered Salmonella cells went from a prestimulus behavior of 14% clockwise rotation to 80% clockwise rotation when 40 mM acetate was added and remained this way for more than 30 min. A low external pH in the absence of weak acid did not markedly affect steady-state tumbling frequency. Among the weak acids tested, the rank for acidity (salicylate greater than benzoate greater than acetate greater than 5,5-dimethyl-2,4-oxazolidinedione) was the same as the rank for the ability to collapse the transmembrane pH gradient and to cause tumbling. At pH 7.0, the tumbling responses caused by the weak acids were much briefer. Indole, a non-weak-acid repellent, did not cause prolonged tumbling at low pH. Two chemotaxis mutants (a Salmonella mutant defective in the chemotaxis methylesterase and an E. coli mutant defective in the methyl-accepting protein in MCP I) showed inverse responses of enhanced counterclockwise rotation in the first 1 min after acetate addition. The latter mutant had been found previously to be defective in the sensing of gradients of extracellular pH and (at neutral pH) of acetate. We conclude (i) that taxes away from acid pH and membrane-permeant weak acids are both mediated by a pH-sensitive component located either in the cytoplasm or on the cytoplasmic side of the membrane, rather than by an external receptor (as in the case of the attractants), and (ii) that both of these taxes involve components of the chemotaxis methylation system, at least in the early phase of the response.

  15. Extracellular vesicles for nucleic acid delivery: progress and prospects for safe RNA-based gene therapy.

    PubMed

    Jiang, L; Vader, P; Schiffelers, R M

    2017-03-01

    Nucleic acid-based drugs offer a potentially effective tool for treatment of a variety of diseases, including cancer, cardiovascular diseases, neurological disorders and infectious diseases. However, clinical applications are hindered by instability of RNA molecules in the circulation and lack of efficient vectors that can deliver RNAs to target tissues and into diseased target cells. Synthetic polymer and lipids as well as virus-based vectors are among the most widely explored vehicles for RNA delivery, but clinical progress has been limited as a result of issues related to toxicity, immunogenicity and low efficiency. Most recently, the discovery that extracellular vesicles (EVs) are endogenous RNA carriers, which may display better biocompatibility and higher delivery efficiency as compared with the synthetic systems, has provided a ray of hope in coping with the delivery dilemma, and EV-based gene therapy has already sparked general interest both in academia and industry. In this review, the current knowledge on EV biology and their role in cell-cell communication will be summarized. Promises of EVs as drug carriers and recent technologies on tailoring EVs' biological attributes will be included, and preclinical studies in which EVs have shown promise for therapeutic RNA delivery will be discussed.

  16. Liposomal delivery of hydrophobic weak acids: enhancement of drug retention using a high intraliposomal pH.

    PubMed

    Joguparthi, Vijay; Anderson, Bradley D

    2008-01-01

    Clinical development of highly potent lipophilic neutral camptothecins has been impeded by the poor solubility, stability, and nonspecific toxicity of these compounds. Liposomal encapsulation offers a promising formulation route for tumor site-specific delivery of these novel drug candidates. However, the development of formulation strategies for liposomal loading and retention of hydrophobic drugs such as the neutral camptothecins has been lacking. In the studies presented here, we explored the potential of a trans-bilayer pH gradient strategy for prolonging the liposome retention of DB-67, a novel lipophilic camptothecin that can undergo lactone ring-opening to form a hydrophobic weak acid. The liposome membrane permeability of DB-67 was obtained as a function of pH in aqueous buffers. A permeability model was developed and liposome membrane permeability was shown to be controlled by the fraction of unbound neutral lactone entrapped in the vesicles. Liposome membrane permeability of DB-67 was also studied under physiological conditions. The high membrane partitioning of DB-67 in the intraliposomal microenvironment was found to shift the equilibrium between lactone and carboxylate towards the lactone species resulting in a faster than desired drug release under physiological conditions. The effectiveness of the pH gradient strategy was further reduced under physiological conditions by the rapid loss of trans-membrane pH gradients due to CO(2) uptake. Simulations were conducted to explore the role of membrane binding, intravesicular pH, and carbonate buffer concentration in successful utilization of the pH gradient strategy for hydrophobic weak acids. (c) 2007 Wiley-Liss, Inc.

  17. Effect of systemic pH on pH sub i and lactic acid generation in exhaustive forearm exercise

    SciTech Connect

    Hood, V.L.; Schubert, C.; Keller, U.; Mueller, S. Univ. of Vermont College of Medicine, Burlington )

    1988-09-01

    To investigate whether changes in systemic pH affect intracellular pH (pH{sub i}), energy-rich phosphates, and lactic acid generation in muscle, eight normal volunteers performed exhaustive forearm exercise with arterial blood flow occluded for 2 min on three occasions. Subjects ingested 4 mmol/kg NH{sub 4}Cl (acidosis; A) or NaHCO{sub 3} (alkalosis; B) or nothing (control; C) 3 h before the exercise. Muscle pH{sub i} and phosphocreatine (PCr) content were measured with {sup 31}P-nuclear magnetic resonance ({sup 31}P-NMR) spectroscopy during exercise and recovery. Lactate output during 0.5-7 min of recovery was calculated as deep venous-arterial concentration differences times forearm blood flow. Before exercise, blood pH and bicarbonate were lower in acidosis than alkalosis and intermediate in control. Lactic acid output during recovery was less with A than B and intermediate in C. PCr utilization and resynthesis were not affected by extracellular pH changes. pH{sub i} did not differ before exercise or at its end. Hence systemic acidosis inhibited and alkalosis stimulated lactic acid output. These findings suggest that systemic pH regulates cellular acid production, protecting muscle pH, at the expense of energy availability.

  18. Electron spin echo modulation studies of doxylstearic acid spin probes in frozen vesicles: Interaction of the spin probe with D sub 2 O and effects of cholesterol addition

    SciTech Connect

    Hiff, T.; Kevan, L. )

    1989-02-23

    Electron spin echo studies have been carried out for a series of x-doxylstearic acid (x = 5, 7, 10, 12 and 16) spin probes in frozen deuteriated aqueous solutions of phospholipid vesicles and cationic dioctadecyldimethylammonium chloride (DODAC) vesicles. Modulation effects due to interactions of the nitroxide group of the spin probes with D{sub 2}O give information about the conformations of the probes and the degree of hydration of the surfactant headgroups as well as about the degree of packing of the alkyl chain. We show that DODAC headgroups are more hydrated than choline headgroups and that the doxylstearic acid probes show a larger tendency for bending in DODAC vesicles than in phospholipid vesicles. Upon addition of cholesterol into phospholipid vesicles, the headgroups are separated and their degree of hydration increases.

  19. L-lysine uptake in giant vesicles from cardiac ventricular sarcolemma: two components of cationic amino acid transport.

    PubMed

    Lu, Xiaodong; Zheng, Ruifang; Gonzalez, Jorge; Gaspers, Lawrence; Kuzhikandathil, Eldo; Peluffo, R Daniel

    2009-08-01

    Cationic L-amino acids enter cardiac-muscle cells through carrier-mediated transport. To study this process in detail, L-[(14)C]lysine uptake experiments were conducted within a 10(3)-fold range of L-lysine concentrations in giant sarcolemmal vesicles prepared from rat cardiac ventricles. Vesicles had a surface-to-volume ratio comparable with that of an epithelial cell, thus representing a suitable system for initial uptake rate studies. Two Na(+)-independent, N-ethylmaleimide-sensitive uptake components were found, one with high apparent affinity (K(m)=222+/-71 microM) and low transport capacity (V(max)=121+/-36 pmol/min per mg of vesicle protein) and the other with low apparent affinity (K(m)=16+/-4 mM) and high capacity (V(max)=4.0+/-0.4 nmol/min per mg of vesicle protein). L-Lysine uptake mediated by both components was stimulated by the presence of intravesicular L-lysine as well as by valinomycin-induced membrane hyperpolarization. Altogether, this behaviour is consistent with the functional properties of the CAT-1 and CAT-2A members of the system y(+) family of cationic amino acid transporters. Furthermore, mRNA transcripts for these two carrier proteins were identified in freshly isolated rat cardiac myocytes, the amount of CAT-1 mRNA, relative to beta-actin, being 33-fold larger than that of CAT-2A. These two transporters appear to function simultaneously as a homoeostatic device that supplies cardiac-muscle cells with cationic amino acids under a variety of metabolic conditions. Analysis of two carriers acting in parallel with such an array of kinetic parameters shows significant activity of the low-affinity component even at amino acid plasma levels far below its K(m).

  20. Effects of pH adjustment and sodium ions on sour taste intensity of organic acids

    USDA-ARS?s Scientific Manuscript database

    Protonated organic acid species have been shown to be the primary stimuli responsible for sour taste of organic acids. However, we have observed that sour taste may be modulated when the pH of acid solutions is raised using sodium hydroxide. Objectives were to evaluate the effect of pH adjustment on...

  1. Interpretation of pH, acidity, and alkalinity in fisheries and aquaculture

    USDA-ARS?s Scientific Manuscript database

    Measurements of pH, acidity, and alkalinity are commonly used to describe water quality. The three variables are interrelated and are sometimes confused. The pH of water is an intensity factor, while the acidity and alkalinity of waters are capacity factors. More precisely, acidity and alkalinity ar...

  2. Single-vesicle imaging reveals different transport mechanisms between glutamatergic and GABAergic vesicles.

    PubMed

    Farsi, Zohreh; Preobraschenski, Julia; van den Bogaart, Geert; Riedel, Dietmar; Jahn, Reinhard; Woehler, Andrew

    2016-02-26

    Synaptic transmission is mediated by the release of neurotransmitters, which involves exo-endocytotic cycling of synaptic vesicles. To maintain synaptic function, synaptic vesicles are refilled with thousands of neurotransmitter molecules within seconds after endocytosis, using the energy provided by an electrochemical proton gradient. However, it is unclear how transmitter molecules carrying different net charges can be efficiently sequestered while maintaining charge neutrality and osmotic balance. We used single-vesicle imaging to monitor pH and electrical gradients and directly showed different uptake mechanisms for glutamate and γ-aminobutyric acid (GABA) operating in parallel. In contrast to glutamate, GABA was exchanged for protons, with no other ions participating in the transport cycle. Thus, only a few components are needed to guarantee reliable vesicle filling with different neurotransmitters. Copyright © 2016, American Association for the Advancement of Science.

  3. High perfluorooctanoic acid exposure induces autophagy blockage and disturbs intracellular vesicle fusion in the liver.

    PubMed

    Yan, Shengmin; Zhang, Hongxia; Guo, Xuejiang; Wang, Jianshe; Dai, Jiayin

    2017-01-01

    Perfluorooctanoic acid (PFOA) has been shown to cause hepatotoxicity and other toxicological effects. Though PPARα activation by PFOA in the liver has been well accepted as an important mechanism of PFOA-induced hepatotoxicity, several pieces of evidence have shown that the hepatotoxic effects of PFOA may not be fully explained by PPARα activation. In this study, we observed autophagosome accumulation in mouse livers as well as HepG2 cells after PFOA exposure. Further in vitro study revealed that the accumulation of autophagosomes was not caused by autophagic flux stimulation. In addition, we observed that PFOA exposure affected the proteolytic activity of HepG2 cells while significant dysfunction of lysosomes was not detected. Quantitative proteomic analysis of crude lysosomal fractions from HepG2 cells treated with PFOA revealed that 54 differentially expressed proteins were related to autophagy or vesicular trafficking and fusion. The proteomic results were further validated in the cells in vitro and livers in vivo after PFOA exposure, which implied potential dysfunction at the late stage of autophagy. However, in HepG2 cells, it seemed that further inhibition of autophagy did not significantly alter the effects of PFOA on cell viability. Although these findings demonstrate that PFOA blocked autophagy and disturbed intracellular vesicle fusion in the liver, the changes in autophagy were observed only at high cytotoxic concentrations of PFOA, suggesting that autophagy may not be a primary target or mode of toxicity. Furthermore, since altered liver autophagy was not observed at concentrations of PFOA associated with human exposures, the relevance of these findings must be questioned.

  4. Distinct changes in neuronal and astrocytic amino acid neurotransmitter metabolism in mice with reduced numbers of synaptic vesicles.

    PubMed

    Bogen, Inger Lise; Risa, Øystein; Haug, Kristin H; Sonnewald, Ursula; Fonnum, Frode; Walaas, S Ivar

    2008-06-01

    The relations between glutamate and GABA concentrations and synaptic vesicle density in nerve terminals were examined in an animal model with 40-50% reduction in synaptic vesicle numbers caused by inactivation of the genes encoding synapsin I and II. Concentrations and synthesis of amino acids were measured in extracts from cerebrum and a crude synaptosomal fraction by HPLC and (13)C nuclear magnetic resonance spectroscopy (NMRS), respectively. Analysis of cerebrum extracts, comprising both neurotransmitter and metabolic pools, showed decreased concentration of GABA, increased concentration of glutamine and unchanged concentration of glutamate in synapsin I and II double knockout (DKO) mice. In contrast, both glutamate and GABA concentrations were decreased in crude synaptosomes isolated from synapsin DKO mice, suggesting that the large metabolic pool of glutamate in the cerebral extracts may overshadow minor changes in the transmitter pool. (13)C NMRS studies showed that the changes in amino acid concentrations in the synapsin DKO mice were caused by decreased synthesis of GABA (20-24%) in cerebral neurons and increased synthesis of glutamine (36%) in astrocytes. In a crude synaptosomal fraction, the glutamate synthesis was reduced (24%), but this reduction could not be detected in cerebrum extracts. We suggest that lack of synaptic vesicles causes down-regulation of neuronal GABA and glutamate synthesis, with a concomitant increase in astrocytic synthesis of glutamine, in order to maintain normal neurotransmitter concentrations in the nerve terminal cytosol.

  5. The PH gene determines fruit acidity and contributes to the evolution of sweet melons.

    PubMed

    Cohen, Shahar; Itkin, Maxim; Yeselson, Yelena; Tzuri, Galil; Portnoy, Vitaly; Harel-Baja, Rotem; Lev, Shery; Sa'ar, Uzi; Davidovitz-Rikanati, Rachel; Baranes, Nadine; Bar, Einat; Wolf, Dalia; Petreikov, Marina; Shen, Shmuel; Ben-Dor, Shifra; Rogachev, Ilana; Aharoni, Asaph; Ast, Tslil; Schuldiner, Maya; Belausov, Eduard; Eshed, Ravit; Ophir, Ron; Sherman, Amir; Frei, Benedikt; Neuhaus, H Ekkehard; Xu, Yimin; Fei, Zhangjun; Giovannoni, Jim; Lewinsohn, Efraim; Tadmor, Yaakov; Paris, Harry S; Katzir, Nurit; Burger, Yosef; Schaffer, Arthur A

    2014-06-05

    Taste has been the subject of human selection in the evolution of agricultural crops, and acidity is one of the three major components of fleshy fruit taste, together with sugars and volatile flavour compounds. We identify a family of plant-specific genes with a major effect on fruit acidity by map-based cloning of C. melo PH gene (CmPH) from melon, Cucumis melo taking advantage of the novel natural genetic variation for both high and low fruit acidity in this species. Functional silencing of orthologous PH genes in two distantly related plant families, cucumber and tomato, produced low-acid, bland tasting fruit, showing that PH genes control fruit acidity across plant families. A four amino-acid duplication in CmPH distinguishes between primitive acidic varieties and modern dessert melons. This fortuitous mutation served as a preadaptive antecedent to the development of sweet melon cultigens in Central Asia over 1,000 years ago.

  6. Acidic pH promotes intervertebral disc degeneration: Acid-sensing ion channel -3 as a potential therapeutic target

    PubMed Central

    Gilbert, Hamish T. J.; Hodson, Nathan; Baird, Pauline; Richardson, Stephen M.; Hoyland, Judith A.

    2016-01-01

    The aetiology of intervertebral disc (IVD) degeneration remains poorly understood. Painful IVD degeneration is associated with an acidic intradiscal pH but the response of NP cells to this aberrant microenvironmental factor remains to be fully characterised. The aim here was to address the hypothesis that acidic pH, similar to that found in degenerate IVDs, leads to the altered cell/functional phenotype observed during IVD degeneration, and to investigate the involvement of acid-sensing ion channel (ASIC) -3 in the response. Human NP cells were treated with a range of pH, from that of a non-degenerate (pH 7.4 and 7.1) through to mildly degenerate (pH 6.8) and severely degenerate IVD (pH 6.5 and 6.2). Increasing acidity of pH caused a decrease in cell proliferation and viability, a shift towards matrix catabolism and increased expression of proinflammatory cytokines and pain-related factors. Acidic pH resulted in an increase in ASIC-3 expression. Importantly, inhibition of ASIC-3 prevented the acidic pH induced proinflammatory and pain-related phenotype in NP cells. Acidic pH causes a catabolic and degenerate phenotype in NP cells which is inhibited by blocking ASIC-3 activity, suggesting that this may be a useful therapeutic target for treatment of IVD degeneration. PMID:27853274

  7. Light-Activated Amino Acid Transport Systems in Halobacterium halobium Envelope Vesicles: Role of Chemical and Electrical Gradients

    NASA Technical Reports Server (NTRS)

    MacDonald, Russell E.; Greene, Richard V.; Lanyi, Janos K.

    1977-01-01

    The accumulation of 20 commonly occurring L-amino acids by cell envelope vesicles of Halobacterium halobium, in response to light-induced membrane potential and an artificially created sodium gradient, has been studied. Nineteen of these amino acids are actively accumulated under either or both of these conditions. Glutamate is unique in that its uptake is driven only by a chemical gradient for sodium. Amino acid concentrations at half-maximal uptake rates (Km) and maximal transport rates (V(sub max) have been determined for the uptake of all 19 amino acids. The transport systems have been partially characterized with respect to groups of amino acids transported by common carriers, cation effects, and relative response to the electrical and chemical components of the sodium gradient, the driving forces for uptake. The data presented clearly show that the carrier systems, which are responsible for uptake of individual amino acids, are as variable in their properties as those found in other organisms, i. e., some are highly specific for individual amino acids, some transport several amino acids competitively, some are activated by a chemical gradient of sodium only, and some function also in the complete absence of such a gradient. For all amino acids, Na(+) and K(+) are both required for maximal rate of uptake. The carriers for L-leucine and L-histidine are symmetrical in that these amino acids are transported in both directions across the vesicle membrane. It is suggested that coupling of substrate transport to metabolic energy via transient ionic gradients may be a general phenomenon in procaryotes.

  8. Light-Activated Amino Acid Transport Systems in Halobacterium halobium Envelope Vesicles: Role of Chemical and Electrical Gradients

    NASA Technical Reports Server (NTRS)

    MacDonald, Russell E.; Greene, Richard V.; Lanyi, Janos K.

    1977-01-01

    The accumulation of 20 commonly occurring L-amino acids by cell envelope vesicles of Halobacterium halobium, in response to light-induced membrane potential and an artificially created sodium gradient, has been studied. Nineteen of these amino acids are actively accumulated under either or both of these conditions. Glutamate is unique in that its uptake is driven only by a chemical gradient for sodium. Amino acid concentrations at half-maximal uptake rates (Km) and maximal transport rates (V(sub max) have been determined for the uptake of all 19 amino acids. The transport systems have been partially characterized with respect to groups of amino acids transported by common carriers, cation effects, and relative response to the electrical and chemical components of the sodium gradient, the driving forces for uptake. The data presented clearly show that the carrier systems, which are responsible for uptake of individual amino acids, are as variable in their properties as those found in other organisms, i. e., some are highly specific for individual amino acids, some transport several amino acids competitively, some are activated by a chemical gradient of sodium only, and some function also in the complete absence of such a gradient. For all amino acids, Na(+) and K(+) are both required for maximal rate of uptake. The carriers for L-leucine and L-histidine are symmetrical in that these amino acids are transported in both directions across the vesicle membrane. It is suggested that coupling of substrate transport to metabolic energy via transient ionic gradients may be a general phenomenon in procaryotes.

  9. A two-component micelle with emergent pH responsiveness by mixing dilauroyl phosphocholine and deoxycholic acid and its delivery of proteins into the cytosol.

    PubMed

    Miyamoto, Noriko; Fujii, Shota; Mochizuki, Shinichi; Sakurai, Kazuo; Sakaguchi, Naoki; Koiwai, Kazunori

    2017-06-01

    Providing appropriate pH responsiveness for drug delivery nanoparticles is one of the major issues in developing a new generation of delivery systems. This paper reports that, when phosphocholine and a bile acid were mixed, the resultant two-component micelle gained pH responsiveness, while the individual components did not show any such responsiveness. The pH responsiveness was shown to be determined by the chemical structure, especially the positions and chirality of the OH groups, of the bile acid, and the sensitivity was determined by the alkyl chain length of the phosphocholine. The best combination for evading endocytosis was dilauroyl phosphocholine (DLPC) and deoxycholic acid (DA). Small-angle X-ray scattering revealed that the pH responsiveness was related to the change of surface hydrophobicity, namely, decreasing pH led to protonation of the carboxylic acid, resulting in aggregation of the preceding micelles. We assume that particles that become hydrophobic in this way can start interacting with the endocytotic bilayer, which eventually leads to rupture of the endocytotic vesicle. This mechanism is well supported by the finding that fluorescein-conjugated ovalbumin proteins were transported into the cytosol when they were co-administered with DLPC/DA. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Vesicle Photonics

    SciTech Connect

    Vasdekis, Andreas E.; Scott, E. A.; Roke, Sylvie; Hubbell, J. A.; Psaltis, D.

    2013-04-03

    Thin membranes, under appropriate boundary conditions, can self-assemble into vesicles, nanoscale bubbles that encapsulate and hence protect or transport molecular payloads. In this paper, we review the types and applications of light fields interacting with vesicles. By encapsulating light-emitting molecules (e.g. dyes, fluorescent proteins, or quantum dots), vesicles can act as particles and imaging agents. Vesicle imaging can take place also under second harmonic generation from vesicle membrane, as well as employing mass spectrometry. Light fields can also be employed to transport vesicles using optical tweezers (photon momentum) or directly pertrurbe the stability of vesicles and hence trigger the delivery of the encapsulated payload (photon energy).

  11. Transport of amino acids (L-valine, L-lysine, L-glutamic acid) and sucrose into plasma membrane vesicles isolated from cotyledons of developing pea seeds.

    PubMed

    de Jong, A; Borstlap, A C

    2000-10-01

    Transport of the amino acids L-valine, L-lysine, and L-glutamic acid and of sucrose was studied in plasma membrane vesicles isolated from developing cotyledons of pea (Pisum sativum L. cv. Marzia). The vesicles were obtained by aqueous polymer two-phase partitioning of a microsomal fraction and the uptake was determined after the imposition of a H(+)-gradient (DeltapH, inside alkaline) and/or an electrical gradient (Deltapsi, inside negative) across the vesicle membrane. In the absence of gradients, a distinct, time-dependent uptake of L-valine was measured, which could be enhanced about 2-fold by the imposition of DeltapH. The imposition of Deltapsi stimulated the influx of valine by 20%, both in the absence and in the presence of DeltapH. Uptake of L-lysine was more strongly stimulated by Deltapsi than by DeltapH, and its DeltapH-dependent uptake was enhanced about 6-fold by the simultaneous imposition of Deltapsi. In the absence of gradients the uptake of L-glutamic acid was about 2-fold higher than that of L-valine, but it was not detectably affected by DeltapH or Deltapsi. Although the transport of sucrose was very low, a stimulating effect of DeltapH could be clearly demonstrated. The results lend further support to the contention that during seed development cotyledonary cells employ H(+)-symporters for the active uptake of sucrose and amino acids.

  12. Organic Acid Excretion in Penicillium ochrochloron Increases with Ambient pH

    PubMed Central

    Vrabl, Pamela; Fuchs, Viktoria; Pichler, Barbara; Schinagl, Christoph W.; Burgstaller, Wolfgang

    2012-01-01

    Despite being of high biotechnological relevance, many aspects of organic acid excretion in filamentous fungi like the influence of ambient pH are still insufficiently understood. While the excretion of an individual organic acid may peak at a certain pH value, the few available studies investigating a broader range of organic acids indicate that total organic acid excretion rises with increasing external pH. We hypothesized that this phenomenon might be a general response of filamentous fungi to increased ambient pH. If this is the case, the observation should be widely independent of the organism, growth conditions, or experimental design and might therefore be a crucial key point in understanding the function and mechanisms of organic acid excretion in filamentous fungi. In this study we explored this hypothesis using ammonium-limited chemostat cultivations (pH 2–7), and ammonium or phosphate-limited bioreactor batch cultivations (pH 5 and 7). Two strains of Penicillium ochrochloron were investigated differing in the spectrum of excreted organic acids. Confirming our hypothesis, the main result demonstrated that organic acid excretion in P. ochrochloron was enhanced at high external pH levels compared to low pH levels independent of the tested strain, nutrient limitation, and cultivation method. We discuss these findings against the background of three hypotheses explaining organic acid excretion in filamentous fungi, i.e., overflow metabolism, charge balance, and aggressive acidification hypothesis. PMID:22493592

  13. Effects of pH and Temperature on the Fatty Acid Composition of Bacillus acidocaldarius

    PubMed Central

    De Rosa, M.; Gambacorta, A.; Bu'lock, J. D.

    1974-01-01

    The fatty acid composition of lipid extracts from cells of Bacillus acidocaldarius grown at temperatures of 50 to 70 C and pH values of 2 to 5 was determined by gas chromatography of the methyl esters. The most abundant fatty acids are 11-cyclohexylundecanoic and 13-cyclohexyltridecanoic, followed by anteiso- and iso-heptadecanoic; unsaturated acids are absent. Highly aerated cultures produce more of the iso and anteiso acids and less of the cyclohexyl acids. The effects of temperature and pH are interdependent; at lower pH, increasing temperature raises the proportion of the iso and anteiso acids, but at higher pH the effect of increasing temperature is reversed and the proportion of the cyclohexyl acids is increased. PMID:4808902

  14. Combined impact of pH and organic acids on iron uptake by Caco-2 cells.

    PubMed

    Salovaara, Susan; Sandberg, Ann-Sofie; Andlid, Thomas

    2003-12-17

    Previous studies have shown that organic acids have an impact on both Fe(II) and Fe(III) uptake in Caco-2 cell. However, to what extent this effect is correlated with the anion of organic acids per se, or with the resulting decrease in pH, has not yet been clarified. Therefore, we studied the effect of five organic acids (tartaric, succinic, citric, oxalic, and propionic acid) on the absorption of Fe(II) and Fe(III) in Caco-2 cells and compared this with sample solutions without organic acids but set to equivalent pH by HCl. The results showed that the mechanisms behind the enhancing effect of organic acids differed for the two forms of iron. For ferric iron the organic acids promoted uptake both by chelation and by lowering the pH, whereas for ferrous iron the promoting effect was caused only by the lowered pH.

  15. Isolation of Plasma Membrane Vesicles from Mouse Placenta at Term and Measurement of System A and System β Amino Acid Transporter Activity

    PubMed Central

    Kusinski, L.C.; Jones, C.J.P.; Baker, P.N.; Sibley, C.P.; Glazier, J.D.

    2010-01-01

    Placental amino acid transport is essential for optimal fetal growth and development, with a reduced fetal provision of amino acids being implicated as a potential cause of fetal growth restriction (FGR). Understanding placental insufficiency related FGR has been aided by the development of mouse models that have features of the human disease. However, to take maximal advantage of these, methods are required to study placental function in the mouse. Here, we report a method to isolate plasma membrane vesicles from mouse placenta near-term and have used these to investigate two amino acid transporters, systems A and β, the activities of which are reduced in human placental microvillous plasma membrane (MVM) vesicles from FGR pregnancies. Plasma membrane vesicles were isolated at embryonic day 18 by a protocol involving homogenisation, MgCl2 precipitation and centrifugation. Vesicles were enriched 11.3 ± 0.5-fold in alkaline phosphatase activity as compared to initial homogenate, with minimal intracellular organelle contamination as judged by marker analyses. Cytochemistry revealed alkaline phosphatase was localised between trophoblast layers I and II, with intense reaction product deposited on the maternal-facing plasma membrane of layer II, suggesting that vesicles were derived from this trophoblast membrane. System A and system β activity in mouse placental vesicles, measured as Na+-dependent uptake of 14C-methylaminoisobutyric acid (MeAIB) and 3H-taurine respectively confirmed localisation of these transporters to the maternal-facing plasma membrane of layer II. Comparison to human placental MVM showed that system A activity was comparable at initial rate between species whilst system β activity was significantly lower in mouse. This mirrored the lower expression of TAUT observed in mouse placental vesicles. We conclude that syncytiotrophoblast layer II-derived plasma membrane vesicles can be isolated and used to examine transporter function. PMID:19954844

  16. Rhizosphere pH responses to simulated acid rain as measured with glass microelectrodes

    SciTech Connect

    Conkling, B.L.

    1988-01-01

    The objectives of this study were to develop a useful experimental system for studying the rhizosphere of growing roots, and to investigate the effects of bulk soil pH and foliar acid rain application on the rhizosphere pH of alfalfa, corn and soybeans. First, a study was done to compare soil pH measurements made with a standard glass pH electrode with those made using an antimony (Sb) microelectrode. Because of uncertainty with the Sb microelectrodes' response, glass pH-sensitive microelectrodes were made and tested for rhizosphere pH measurements. The influence of soil water pressure gradients in the range of {minus}10 to {minus}1500 kPa in the proximity of the pH and reference electrodes on pH measurements made with microelectrodes was studied. The effect of foliar acid rain application on the rhizosphere pH of alfalfa, corn, and soybean as a function of soil pH were studied. Alfalfa, corn, and soybean were grown into minirhizotrons containing reformed samples of both Seymour A and Bt soil horizons, and the rhizosphere pH measured. The measured in situ bulk soil pH ranged from 4.9 to 6.2 in the A horizon and from 4.0 to 5.7 in the Bt horizon. Plants received acid or non-acid foliar rain applications. Rhizosphere pH was measured using a glass pH-sensitive microelectrode. Acid rain applications caused foliar damage, but had little effect on the rhizosphere pH. The general trend was for the lateral root pH values to be slightly higher than the main root values.

  17. Acidic Extracellular pH Promotes Activation of Integrin αvβ3

    PubMed Central

    Paradise, Ranjani K.; Lauffenburger, Douglas A.; Van Vliet, Krystyn J.

    2011-01-01

    Acidic extracellular pH is characteristic of the cell microenvironment in several important physiological and pathological contexts. Although it is well established that acidic extracellular pH can have profound effects on processes such as cell adhesion and migration, the underlying molecular mechanisms are largely unknown. Integrin receptors physically connect cells to the extracellular matrix, and are thus likely to modulate cell responses to extracellular conditions. Here, we examine the role of acidic extracellular pH in regulating activation of integrin αvβ3. Through computational molecular dynamics simulations, we find that acidic extracellular pH promotes opening of the αvβ3 headpiece, indicating that acidic pH can thereby facilitate integrin activation. This prediction is consistent with our flow cytometry and atomic force microscope-mediated force spectroscopy assays of integrin αvβ3 on live cells, which both demonstrate that acidic pH promotes activation at the intact cell surface. Finally, quantification of cell morphology and migration measurements shows that acidic extracellular pH affects cell behavior in a manner that is consistent with increased integrin activation. Taken together, these computational and experimental results suggest a new and complementary mechanism of integrin activation regulation, with associated implications for cell adhesion and migration in regions of altered pH that are relevant to wound healing and cancer. PMID:21283814

  18. Weak-acid preservatives: pH and proton movements in the yeast Saccharomyces cerevisiae.

    PubMed

    Stratford, Malcolm; Nebe-von-Caron, Gerhard; Steels, Hazel; Novodvorska, Michaela; Ueckert, Joerg; Archer, David B

    2013-02-15

    Weak-acid preservatives commonly used to prevent fungal spoilage of low pH foods include sorbic and acetic acids. The "classical weak-acid theory" proposes that weak acids inhibit spoilage organisms by diffusion of undissociated acids through the membrane, dissociation within the cell to protons and anions, and consequent acidification of the cytoplasm. Results from 25 strains of Saccharomyces cerevisiae confirmed inhibition by acetic acid at a molar concentration 42 times higher than sorbic acid, in contradiction of the weak-acid theory where all acids of equal pK(a) should inhibit at equimolar concentrations. Flow cytometry showed that the intracellular pH fell to pH 4.7 at the growth-inhibitory concentration of acetic acid, whereas at the inhibitory concentration of sorbic acid, the pH only fell to pH 6.3. The plasma membrane H⁺-ATPase proton pump (Pma1p) was strongly inhibited by sorbic acid at the growth-inhibitory concentration, but was stimulated by acetic acid. The H⁺-ATPase was also inhibited by lower sorbic acid concentrations, but later showed recovery and elevated activity if the sorbic acid was removed. Levels of PMA1 transcripts increased briefly following sorbic acid addition, but soon returned to normal levels. It was concluded that acetic acid inhibition of S. cerevisiae was due to intracellular acidification, in accord with the "classical weak-acid theory". Sorbic acid, however, appeared to be a membrane-active antimicrobial compound, with the plasma membrane H⁺-ATPase proton pump being a primary target of inhibition. Understanding the mechanism of action of sorbic acid will hopefully lead to improved methods of food preservation. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Negative pH and extremely acidic mine waters from Iron Mountain, California

    USGS Publications Warehouse

    Nordstrom, D.K.; Alpers, C.N.; Ptacek, C.J.; Blowes, D.W.

    2000-01-01

    Extremely acidic mine waters with pH values as low as -3.6, total dissolved metal concentrations as high as 200 g/L, and sulfate concentrations as high as 760 g/L, have been encountered underground in the Richmond Mine at Iron Mountain, CA. These are the most acidic waters known. The pH measurements were obtained by using the Pitzer method to define pH for calibration of glass membrane electrodes. The calibration of pH below 0.5 with glass membrane electrodes becomes strongly nonlinear but is reproducible to a pH as low as -4. Numerous efflorescent minerals were found forming from these acid waters. These extreme acid waters were formed primarily by pyrite oxidation and concentration by evaporation with minor effects from aqueous ferrous iron oxidation and efflorescent mineral formation.

  20. Artificially Smart Vesicles with Superior Structural Stability: Fabrication, Characterizations, and Transmembrane Traffic.

    PubMed

    Zhang, Wen-Jian; Hong, Chun-Yan; Pan, Cai-Yuan

    2017-05-03

    Intelligent vesicles are fabricated at up to 30% solid content via an approach of polymerization-induced self-assembly and reorganization (PISR). Upon irradiation with UV light (365 nm), light-triggered dimerization of the coumarin moieties anchored in the membrane leads to the membrane cross-linking of the vesicles, which endows the vesicles with superior structural stability. Due to the tertiary amine groups in the membrane, the vesicles go through a swelling/deswelling change upon switching the pH values. In acidic aqueous solution, the pores in the membrane of vesicles are opened, which is beneficial for transmembrane traffic. The pore size in the membrane of vesicles is in accordance with the extent of membrane cross-linking, which can be conveniently regulated by the irradiation time of UV light (365 nm). The size range of the substance for transmembrane traffic is effectively enlarged; even 15 nm gold nanoparticles can be postloaded into the vesicles with lower extents of the membrane cross-linking through the diffusion method. Although the pores in the vesicle membrane are opened in acidic aqueous solution, transmembrane traffic is inhibited for the electropositive substance because of electrostatic repulsion but is allowed for the electronegative substance. These reported vesicles herein may be the smartest artificial vesicles to date due to their multiple selective permeability.

  1. Effect of pH on corrosion inhibition of steel by polyaspartic acid

    SciTech Connect

    Silverman, D.C.; Kalota, D.J.; Stover, F.S.

    1995-10-01

    Polyaspartic acid, a polymeric form of aspartic acid has been examined as a corrosion inhibitor for steel as a function of pH, temperature, and hydrodynamic conditions. The temperature ranged from 25 C to 95 C and the concentration ranged from less than 1% to about 10% by weight. Experimental procedures included electrochemical impedance spectroscopy, the rotating cylinder electrode, and coupon immersion. At low to neutral pH values, polyaspartic acid increases the corrosion rate of steel. At high pH above about 10, polyaspartic acid is a reasonably robust corrosion inhibitor. Between a pH of 7 and 10, corrosion in the presence of polyaspartic acid is a complex function of temperature, concentration, water quality, and hydrodynamic conditions. By combining corrosion potential measurements with speciation diagrams as obtained by titration, a reasonably cohesive explanation of the behavior has been developed.

  2. Effect of pH on corrosion inhibition of steel by polyaspartic acid

    SciTech Connect

    Silverman, D.C.; Kalota, D.J.; Stover, F.S.

    1995-11-01

    Polyaspartic acid, a polymeric form of aspartic acid (C{sub 4}H{sub 7}NO{sub 4}), was examined as a corrosion inhibitor for steel as a function of pH, temperature, and hydrodynamic conditions. The temperature ranged from 25 C to 95 C, and the concentration ranged from < 1 wt% to {approximately} 10 wt%. Experimental procedures included electrochemical impedance spectroscopy (EIS), the rotating cylinder electrode (RCE), and coupon immersion. At low to neutral pH values, polyaspartic acid increased the corrosion rate of steel. At high pH (< {approximately} 10), polyaspartic acid was a reasonably robust corrosion inhibitor. Between pH 7 and 10, corrosion in the presence of polyaspartic acid was a complex function of temperature concentration, water quality, and hydrodynamic conditions. By combining corrosion potential measurements with speciation diagrams obtained by titration, a reasonably cohesive explanation of the behavior was developed.

  3. A structural transition in class II major histocompatibility complex proteins at mildly acidic pH

    PubMed Central

    1996-01-01

    Peptide binding by class II major histocompatibility complex proteins is generally enhanced at low pH in the range of hydrogen ion concentrations found in the endosomal compartments of antigen- presenting cells. We and others have proposed that class II molecules undergo a reversible conformational change at low pH that is associated with enhanced peptide loading. However, no one has previously provided direct evidence for a structural change in class II proteins in the mildly acidic pH conditions in which enhanced peptide binding is observed. In this study, susceptibility to denaturation induced by sodium dodecyl sulfate (SDS) detergent or heat was used to probe the conformation of class II at different hydrogen ion concentrations. Class II molecules became sensitive to denaturation at pH 5.5-6.5 depending on the allele and experimental conditions. The observed structural transition was fully reversible if acidic pH was neutralized before exposure to SDS or heat. Experiments with the environment- sensitive fluorescent probe ANS (8-anilino-1-naphthalene-sulfonic acid) provided further evidence for a reversible structural transition at mildly acidic pH associated with an increase in exposed hydrophobicity in class II molecules. IAd conformation was found to change at a higher pH than IEd, IEk, or IAk, which correlates with the different pH optimal for peptide binding by these molecules. We conclude that pH regulates peptide binding by influencing the structure of class II molecules. PMID:8551215

  4. Continuous intra-arterial blood pH monitoring in rabbits with acid-base disorders.

    PubMed

    Jin, Weizhong; Jiang, Jinjun; Wang, Xun; Zhu, Xiaodan; Wang, Guifang; Song, Yuanlin; Bai, Chunxue

    2011-07-31

    The acid-base balance of arterial blood is important for the clinical management of seriously ill patients, especially patients with acute lung injury or acute respiratory distress syndrome. We developed a novel fluorosensor for continuous blood pH monitoring and evaluated its performance both in vitro and in vivo in rabbits with acid-base disorders. The pH sensor is made of N-allyl-4-piperazinyl-1, 8-napthalimide and 2-hydroxyethyl methacrylate, which were bonded at the distal end of the optical fiber. The fluorescence intensity increased as the pH decreased with good reproducibility, selectivity and linearity in the pH range of 6-8. The pH measurement precision was 0.03 ± 0.03 pH units with a bias of -0.02 ± 0.04 (n = 105) and -0.00 ± 0.05 pH units (n=189) in rabbits with metabolic and respiratory acid-base orders, respectively. The optical pH sensor can accurately measure pH fluctuations with a fast response and is a promising candidate for continuous in-line measurements of blood pH in critical care patients.

  5. Nucleic acid induced unfolding of recombinant prion protein globular fragment is pH dependent.

    PubMed

    Bera, Alakesh; Nandi, Pradip K

    2014-12-01

    Nucleic acid can catalyze the conversion of α-helical cellular prion protein to β-sheet rich Proteinase K resistant prion protein oligomers and amyloid polymers in vitro and in solution. Because unfolding of a protein molecule from its ordered α-helical structure is considered to be a necessary step for the structural conversion to its β-sheet rich isoform, we have studied the unfolding of the α-helical globular 121-231 fragment of mouse recombinant prion protein in the presence of different nucleic acids at neutral and acid pH. Nucleic acids, either single or double stranded, do not have any significant effect on the secondary structure of the protein fragment at neutral pH; however the protein secondary structure is modified by the nucleic acids at pH 5. Nucleic acids do not show any significant effect on the temperature induced unfolding of the globular prion protein domain at neutral pH which, however, undergoes a gross conformational change at pH 5 as evidenced from the lowering of the midpoint of thermal denaturation temperatures, Tm, of the protein. The extent of Tm decrease shows a dependence on the nature of nucleic acid. The interaction of nucleic acid with the nonpolar groups exposed from the protein interior at pH 5 probably contributes substantially to the unfolding process of the protein. © 2014 The Protein Society.

  6. Acidic pH sensing in the bacterial cytoplasm is required for Salmonella virulence.

    PubMed

    Choi, Jeongjoon; Groisman, Eduardo A

    2016-09-01

    pH regulates gene expression, biochemical activities and cellular behaviors. A mildly acidic pH activates the master virulence regulatory system PhoP/PhoQ in the facultative intracellular pathogen Salmonella enterica serovar Typhimurium. The sensor PhoQ harbors an extracytoplasmic domain implicated in signal sensing, and a cytoplasmic domain controlling activation of the regulator PhoP. We now report that, surprisingly, a decrease in Salmonella's own cytoplasmic pH induces transcription of PhoP-activated genes even when the extracytoplasmic pH remains neutral. Amino acid substitutions in PhoQ's cytoplasmic domain hindered activation by acidic pH and attenuated virulence in mice, but did not abolish activation by low Mg(2+) or the antimicrobial peptide C18G. Conversely, removal of PhoQ's extracytoplasmic domains prevented the response to the latter PhoQ-activating signals but not to acidic pH. PhoP-dependent genes were minimally induced by acidic pH in the non-pathogenic species Salmonella bongori but were activated by low Mg(2+) and C18G as in pathogenic S. enterica. Our findings indicate that the sensor PhoQ enables S. enterica to respond to both host- and bacterial-derived signals that alter its cytoplasmic pH. © 2016 John Wiley & Sons Ltd.

  7. A novel acidic pH fluorescent probe based on a benzothiazole derivative.

    PubMed

    Ma, Qiujuan; Li, Xian; Feng, Suxiang; Liang, Beibei; Zhou, Tiqiang; Xu, Min; Ma, Zhuoyi

    2017-04-15

    A novel acidic pH fluorescent probe 1 based on a benzothiazole derivative has been designed, synthesized and developed. The linear response range covers the acidic pH range from 3.44 to 6.46, which is valuable for pH researches in acidic environment. The evaluated pKa value of the probe 1 is 4.23. The fluorescence enhancement of the studied probe 1 with an increase in hydrogen ions concentration is based on the hindering of enhanced photo-induced electron transfer (PET) process. Moreover, the pH sensor possesses a highly selective response to H(+) in the presence of metal ions, anions and other bioactive small molecules which would be interfere with its fluorescent pH response. Furthermore, the probe 1 responds to acidic pH with short response time that was less than 1min. The probe 1 has been successfully applied to confocal fluorescence imaging in live HeLa cells and can selectively stain lysosomes. All of such good properties prove it can be used to monitoring pH fluctuations in acidic environment with high sensitivity, pH dependence and short response time.

  8. A novel acidic pH fluorescent probe based on a benzothiazole derivative

    NASA Astrophysics Data System (ADS)

    Ma, Qiujuan; Li, Xian; Feng, Suxiang; Liang, Beibei; Zhou, Tiqiang; Xu, Min; Ma, Zhuoyi

    2017-04-01

    A novel acidic pH fluorescent probe 1 based on a benzothiazole derivative has been designed, synthesized and developed. The linear response range covers the acidic pH range from 3.44 to 6.46, which is valuable for pH researches in acidic environment. The evaluated pKa value of the probe 1 is 4.23. The fluorescence enhancement of the studied probe 1 with an increase in hydrogen ions concentration is based on the hindering of enhanced photo-induced electron transfer (PET) process. Moreover, the pH sensor possesses a highly selective response to H+ in the presence of metal ions, anions and other bioactive small molecules which would be interfere with its fluorescent pH response. Furthermore, the probe 1 responds to acidic pH with short response time that was less than 1 min. The probe 1 has been successfully applied to confocal fluorescence imaging in live HeLa cells and can selectively stain lysosomes. All of such good properties prove it can be used to monitoring pH fluctuations in acidic environment with high sensitivity, pH dependence and short response time.

  9. Characteristics and anti-proliferative activity of azelaic acid and its derivatives entrapped in bilayer vesicles in cancer cell lines.

    PubMed

    Manosroi, Aranya; Panyosak, Atchara; Rojanasakul, Yon; Manosroi, Jiradej

    2007-06-01

    The hydrophilicity and lipophilicity of azelaic acid (AA) were modified to diethyl azelate (DA) which was synthesized by Fisher esterification reaction and identified by IR, MS and (1)H NMR and to azelaic acid-beta-cyclodextrin complex (AACD) which was prepared by inclusion complexation and identified by IR, DSC and XRD respectively. AA, DA and AACD were entrapped in liposomes and niosomes comprising of L-alpha-dipalmitoyl phosphatidylcholine (DPPC)/cholesterol at 7:3 molar ratio and Tween61/cholesterol at 1:1 molar ratio, respectively, using a thin-film hydration method with sonication. The size and morphology of these bilayer vesicles were determined by optical and transmission electron microscopy. The particle size was found to be in the range of 90-190 nm. The entrapment efficiency of AA, DA and AACD in all vesicular formulations was more than 80%, as analyzed by HPLC for AA and AACD, and GC for DA. Anti-proliferative activity of AA and its derivatives (DA and AACD) both entrapped and not entrapped in bilayer vesicles, using MTT assay in three cancer cell lines (HeLa, KB and B(16)F(10)) comparing with vincristine, were investigated. AACD showed the highest potency comparing to AA in HeLa, KB and B(16)F(10) of 1.48, 1.6 and 1.5 times, respectively. AA entrapped in liposomes was about 90 times more potent than the free AA, and about 1.5 times less potent than vincristine. When entrapped in bilayer vesicles, DA and AACD were more effective than AA in killing cancer cells. AACD entrapped in liposomes gave the highest anti-proliferation activity in HeLa cell lines with the IC(50) of 2.3 and 327 times more potent than vincristine and AA, respectively. DA in liposomes demonstrated the IC(50) of 0.03 times less potent than vincristine in KB cell lines, while in B(16)F(10) AACD in niosomes showed the IC(50) of 0.05 times less potent than vincristine. This study has suggested that the modification of AA by derivatization and complexation as well as the entrapment in

  10. Cut-off net acid generation pH in predicting acid-forming potential in mine spoils.

    PubMed

    Liao, B; Huang, L N; Ye, Z H; Lan, C Y; Shu, W S

    2007-01-01

    Acidification of mine wastes can lead to a series of environmental problems, such as acid drainage, heavy metal mobilization, and ecosystem degradation. Prediction of acid-forming potential is one of the key steps in management of sulfide-bearing mine wastes. In this paper, the acid-forming potential of 180 mine waste samples collected from 17 mine sites in China were studied using a net acid generation (NAG) method. The samples contained different contents of total sulfur (ranging from 0.6 to 200 g kg(-1)), pyritic sulfur (ranging from 0 to 100 g kg(-1)), and acid neutralization capacity (ANC, ranging from -41 to 274 kg H2SO4 t(-1)). Samples with high acid-forming potential are generally due to their high sulfur content or low acid neutralization capacity. After the samples were oxidized by H2O2, the amounts of acid generation and the final NAG pH were measured. Results indicated that the final NAG pH gave a well-defined demarcation between acid-forming and non-acid-forming materials. Samples with final NAG pH >or= 5 could be classified as non-acid-forming materials, while those with NAG pH acid-forming materials. Materials with NAG pH > 2.5, but < 5, had low risk of being acid-forming. The confirmation of cut-off NAG pH will be used as a rapid and cost-effective operational monitoring tool for the in-pit prediction of acid-forming potential of mine wastes and classification of waste types.

  11. Acidic pH of the lateral intercellular spaces of MDCK cells cultured on permeable supports.

    PubMed

    Chatton, J Y; Spring, K R

    1994-06-01

    The pH of the lateral intercellular space (LIS) of Madin-Darby canine kidney (MDCK) cell monolayers grown on permeable supports was investigated by microspectrofluorimetry using BCECF (2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein). The permeability of the support was selectively reduced by growing Zn-Al-silicate crystals inside its pores. The diffusion of BCECF across the filter was sufficiently retarded to allow measurements of fluorescence in the LIS. The LIS pH and intracellular pH of the cells surrounding them were determined in HEPES-buffered solutions. When the perfusate pH was 7.4, the LIS pH was more acidic (7.06 +/- 0.02) and equaled the cytoplasmic pH (7.08 +/- 0.05). When perfusate was changed to pH 7.0 or 7.8, the LIS changed linearly by about half the magnitude of the perfusate pH. Intracellular pH followed LIS pH variations between perfusate pH 7.0 and 7.4 but was significantly higher when perfusate pH was 7.8. Tight junctional H+ permeability was undetectably low. The low steady-state pH in the LIS was not altered by inhibitors of acid transport or low temperature. Rapid perturbations of pH in the LIS showed that protons were not immobilized in the LIS. The acidic microenvironment within the LIS may be the result of buffering by the cell surface proteins.

  12. Effect of initial solution pH on photo-induced reductive decomposition of perfluorooctanoic acid.

    PubMed

    Qu, Yan; Zhang, Chao-Jie; Chen, Pei; Zhou, Qi; Zhang, Wei-Xian

    2014-07-01

    The effects of initial solution pH on the decomposition of perfluorooctanoic acid (PFOA) with hydrated electrons as reductant were investigated. The reductive decomposition of PFOA depends strongly on the solution pH. In the pH range of 5.0-10.0, the decomposition and defluorination rates of PFOA increased with the increase of the initial solution pH. The rate constant was 0.0295 min(-1) at pH 10.0, which was more than 49.0 times higher than that at pH 5.0. Higher pH also inhibits the generation of toxic intermediates during the PFOA decomposition. For example, the short-chain PFCAs reached a lower maximum concentration in shorter reaction time as pH increasing. The peak areas of accumulated fluorinated and iodinated hydrocarbons detected by GC/MS under acidic conditions were nearly 10-100 times more than those under alkaline conditions. In short, alkaline conditions were more favorable for photo-induced reduction of PFOA as high pH promoted the decomposition of PFOA and inhibited the accumulation of intermediate products. The concentration of hydrated electron, detected by laser flash photolysis, increased with the increase of the initial pH. This was the main reason why the decomposition of PFOA in the UV-KI system depended strongly on the initial pH. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Primordial soup or vinaigrette: did the RNA world evolve at acidic pH?

    PubMed Central

    2012-01-01

    Background The RNA world concept has wide, though certainly not unanimous, support within the origin-of-life scientific community. One view is that life may have emerged as early as the Hadean Eon 4.3-3.8 billion years ago with an atmosphere of high CO2 producing an acidic ocean of the order of pH 3.5-6. Compatible with this scenario is the intriguing proposal that life arose within alkaline (pH 9-11) deep-sea hydrothermal vents like those of the 'Lost City', with the interface with the acidic ocean creating a proton gradient sufficient to drive the first metabolism. However, RNA is most stable at pH 4-5 and is unstable at alkaline pH, raising the possibility that RNA may have first arisen in the acidic ocean itself (possibly near an acidic hydrothermal vent), acidic volcanic lake or comet pond. As the Hadean Eon progressed, the ocean pH is inferred to have gradually risen to near neutral as atmospheric CO2 levels decreased. Presentation of the hypothesis We propose that RNA is well suited for a world evolving at acidic pH. This is supported by the enhanced stability at acidic pH of not only the RNA phosphodiester bond but also of the aminoacyl-(t)RNA and peptide bonds. Examples of in vitro-selected ribozymes with activities at acid pH have recently been documented. The subsequent transition to a DNA genome could have been partly driven by the gradual rise in ocean pH, since DNA has greater stability than RNA at alkaline pH, but not at acidic pH. Testing the hypothesis We have proposed mechanisms for two key RNA world activities that are compatible with an acidic milieu: (i) non-enzymatic RNA replication of a hemi-protonated cytosine-rich oligonucleotide, and (ii) specific aminoacylation of tRNA/hairpins through triple helix interactions between the helical aminoacyl stem and a single-stranded aminoacylating ribozyme. Implications of the hypothesis Our hypothesis casts doubt on the hypothesis that RNA evolved in the vicinity of alkaline hydrothermal vents. The

  14. A neutral ceramidase homologue from Dictyostelium discoideum exhibits an acidic pH optimum.

    PubMed Central

    Monjusho, Hatsumi; Okino, Nozomu; Tani, Motohiro; Maeda, Mineko; Yoshida, Motonobu; Ito, Makoto

    2003-01-01

    The nucleotide sequence reported for the Dictyostelium discoideum ceramidase is available on the DNA Data Bank of Japan (DDBJ). Ceramidases (CDases) are currently classified into three categories (acid, neutral and alkaline) based on their optimal pHs and primary structures. Here, we report the first exception to this rule. We cloned the CDase cDNA, consisting of 2142 nucleotides encoding 714 amino-acid residues, from the slime mould, Dictyostelium discoideum. The putative amino-acid sequence indicates 32-42% identity with various neutral CDases, but does not show any similarity to the acid and alkaline CDases, indicating the enzyme should be classified as a neutral CDase. However, overexpression of the cDNA in D. discoideum resulted in increased CDase activity at an acidic, but not a neutral pH range. Knockout of the gene in slime mould eliminated CDase activity at acidic pH. The recombinant enzyme expressed in the slime mould was purified and then characterized. Consequently, the purified CDase was found to exhibit the maximal activity at approx. pH 3.0. The singular pH dependency of slime mould CDase is not derived from the specific post-translational modification in the slime mould, because the enzyme showed an acidic pH optimum even when expressed in Chinese hamster ovary cells, whereas rat neutral-CDase exhibited a neutral pH optimum when expressed in slime mould. PMID:12943537

  15. Formation of elastic whey protein gels at low pH by acid equilibration.

    PubMed

    Vardhanabhuti, Bongkosh; Khayankan, Worarat; Foegeding, E Allen

    2010-06-01

    Whey protein gels have a weak/brittle texture when formed at pH pH is required to produce a high-protein, shelf-stable product. We investigated if gels could be made under conditions that produced strong/elastic textural properties then adjusted to pH pH 7.5). Equilibration in acid solutions caused gel swelling and lowered pH because of the diffusion of water and H(+) into the gels. The type and concentration of acid, and presence of other ions, in the equilibrating solutions influenced pH, swelling ratio, and fracture properties of the gels. Swelling of gels decreased fracture stress (because of decreased protein network density) but caused little change to fracture strain, thus maintaining a desirable strong/elastic fracture pattern. We have shown that whey protein isolate gels can be made at pH acid type, acid concentration, pH of equilibrating solution, and equilibrating time.

  16. Urinary pH reflects dietary acid load in patients with type 2 diabetes.

    PubMed

    Miki, Akane; Hashimoto, Yoshitaka; Tanaka, Muhei; Kobayashi, Yukiko; Wada, Sayori; Kuwahata, Masashi; Kido, Yasuhiro; Yamazaki, Masahiro; Fukui, Michiaki

    2017-07-01

    Dietary acid load is important information, however, survey of food intake needs time and skill. Therefore, it is difficult to survey food intake from all patients. It remains to be elucidated the association between dietary acid load and urinary pH in patients with type 2 diabetes. In this cross-sectional study of 173 patients, we investigated the relationship between urinary pH and dietary acid load, assessed with potential renal acid load. Habitual food and nutrient intake was assessed by a self-administered diet history questionnaire. Urinary pH was negatively correlated with potential renal acid load (r = -0.24, p = 0.002). Multivariate regression analysis revealed that potential renal acid load (standardized regression coefficient = -0.21, p = 0.036) was associated with urinary pH after adjusting for covariates. In addition, according to the receiver operator characteristic analysis, the optimal cut-off point of urinary pH for high dietary acid load, defined as potential renal acid load over 7.0 mEq/day was 5.7 (area under the receiver operator characteristic curve 0.63 (95% CI 0.54-0.71), sensitivity = 0.56, specificity = 0.70, p = 0.004). Urinary pH was associated with dietary acid load in patients with type 2 diabetes. We suggest that urinary pH can be a practical screening marker for dietary acid load in patients with type 2 diabetes.

  17. Urinary pH reflects dietary acid load in patients with type 2 diabetes

    PubMed Central

    Miki, Akane; Hashimoto, Yoshitaka; Tanaka, Muhei; Kobayashi, Yukiko; Wada, Sayori; Kuwahata, Masashi; Kido, Yasuhiro; Yamazaki, Masahiro; Fukui, Michiaki

    2017-01-01

    Dietary acid load is important information, however, survey of food intake needs time and skill. Therefore, it is difficult to survey food intake from all patients. It remains to be elucidated the association between dietary acid load and urinary pH in patients with type 2 diabetes. In this cross-sectional study of 173 patients, we investigated the relationship between urinary pH and dietary acid load, assessed with potential renal acid load. Habitual food and nutrient intake was assessed by a self-administered diet history questionnaire. Urinary pH was negatively correlated with potential renal acid load (r = –0.24, p = 0.002). Multivariate regression analysis revealed that potential renal acid load (standardized regression coefficient = –0.21, p = 0.036) was associated with urinary pH after adjusting for covariates. In addition, according to the receiver operator characteristic analysis, the optimal cut-off point of urinary pH for high dietary acid load, defined as potential renal acid load over 7.0 mEq/day was 5.7 (area under the receiver operator characteristic curve 0.63 (95% CI 0.54–0.71), sensitivity = 0.56, specificity = 0.70, p = 0.004). Urinary pH was associated with dietary acid load in patients with type 2 diabetes. We suggest that urinary pH can be a practical screening marker for dietary acid load in patients with type 2 diabetes. PMID:28751813

  18. The effect of low pH on breast cancer resistance protein (ABCG2)-mediated transport of methotrexate, 7-hydroxymethotrexate, methotrexate diglutamate, folic acid, mitoxantrone, topotecan, and resveratrol in in vitro drug transport models.

    PubMed

    Breedveld, Pauline; Pluim, Dick; Cipriani, Greta; Dahlhaus, Femke; van Eijndhoven, Maria A J; de Wolf, Cornelia J F; Kuil, Annemieke; Beijnen, Jos H; Scheffer, George L; Jansen, Gerrit; Borst, Piet; Schellens, Jan H M

    2007-01-01

    Some cellular uptake systems for (anti)folates function optimally at acidic pH. We have tested whether this also applies to efflux from cells by breast cancer resistance protein (BCRP; ABCG2), which has been reported to transport folic acid, methotrexate, and methotrexate di- and triglutamate at physiological pH. Using Spodoptera frugiperda-BCRP membrane vesicles, we showed that the ATP-dependent vesicular transport of 1 muM methotrexate by BCRP is 5-fold higher at pH 5.5 than at physiological pH. The transport of methotrexate was saturable at pH 5.5, with apparent Km and Vmax values of 1.3 +/- 0.2 mM and 44 +/- 2.5 nmol/mg of protein/min, respectively, but was linear with drug concentration at pH 7.3 up to 6 mM methotrexate. In contrast to recent reports, we did not detect transport of methotrexate diglutamate at physiological pH, but we did find transport at pH 5.5. We also found that 7-hydroxy-methotrexate, the major metabolite of methotrexate, is transported by BCRP both at physiological pH and (more efficiently) at low pH. The pH effect was also observed in intact BCRP-overexpressing cells: we found a 3-fold higher level of resistance to both methotrexate and the prototypical BCRP substrate mitoxantrone at pH 6.5 as at physiological pH. Furthermore, with MDCKII-BCRP monolayers, we found that resveratrol, which is a neutral compound at pH < or = 7.4, is efficiently transported by BCRP at pH 6.0, whereas we did not detect active transport at pH 7.4. We conclude that BCRP transports substrate drugs more efficiently at low pH, independent of the dissociation status of the substrate.

  19. Oxidation in fish oil enriched mayonnaise: ascorbic acid and low pH increase oxidative deterioration.

    PubMed

    Jacobsen, C; Timm, M; Meyer, A S

    2001-08-01

    The effect of ascorbic acid (0-4000 ppm) and pH (3.8-6.2) on oxidation and levels of iron and copper in various fractions of mayonnaise enriched with 16% fish oil was investigated. Ascorbic acid induced release of iron from the assumed oil-water interface into the aqueous phase at all pH levels, but this effect of ascorbic acid was strongest at low pH (pH 3.8-4.2). Ascorbic acid generally promoted formation of volatile oxidation compounds and reduced the peroxide value in mayonnaises. Peroxide values and total volatiles generally increased with decreasing pH values, suggesting that low pH promoted oxidation. It is proposed that iron bridges between the egg yolk proteins low-density lipoproteins, lipovitellin, and phosvitin at the oil-water interface are broken at low pH values, whereby iron ions become accessible as oxidation initiators. In the presence of ascorbic acid, oxidation is further enhanced due to the reduction of Fe(3+) to Fe(2+) that rapidly catalyzes lipid oxidation via lipid hydroperoxide decomposition at the oil-water interface in mayonnaise.

  20. Acid precipitation effects on soil pH and base saturation of exchange sites

    Treesearch

    W. W. McFee; J. M. Kelly; R. H. Beck

    1976-01-01

    The typical values and probable ranges of acid-precipitation are evaluated in terms of their theoretical effects on pH and cation exchange equilibrium of soils characteristic of the humid temperature region. The extent of probable change in soil pH and the time required to cause such a change are calculated for a range of common soils. Hydrogen ion input by acid...

  1. Volatile fatty acids distribution during acidogenesis of algal residues with pH control.

    PubMed

    Li, Yan; Hua, Dongliang; Zhang, Jie; Zhao, Yuxiao; Xu, Haipeng; Liang, Xiaohui; Zhang, Xiaodong

    2013-06-01

    The anaerobic acidification of protein-rich algal residues with pH control (4, 6, 8, 10) was studied in batch reactors, which was operated at mesophilic(35 °C) condition. The distribution of major volatile fatty acids (VFAs) during acidogenesis was emphasized in this paper. The results showed that the acidification efficiency and VFAs distribution in the acid reactor strongly depended on the pH. The main product for all the runs involved acetic acid except that the proportion of butyric acid acidified at pH 6 was relatively higher. The other organic acids remained at lower levels. The VFAs yield reached the maximum value with about 0.6 g VFAs/g volatile solid (VS) added as pH was 8, and also the content of total ammonia nitrogen (TAN) reached the highest values of 9,629 mg/l. Low acidification degrees were obtained under the conditions at pH 4 and 10, which was not suitable for the metabolism of acidogens. Hydralic retention time (HRT) required for different conditions varied. As a consequence, it was indicated that pH was crucial to the acidification efficiency and products distribution. The investigation of acidogenesis process, which was producing the major substrates, short-chain fatty acids, would play the primary role in the efficient operation of methanogenesis.

  2. Change of pH during excess sludge fermentation under alkaline, acidic and neutral conditions.

    PubMed

    Yuan, Yue; Peng, Yongzhen; Liu, Ye; Jin, Baodan; Wang, Bo; Wang, Shuying

    2014-12-01

    The change in pH during excess sludge (ES) fermentation of varying sludge concentrations was investigated in a series of reactors at alkaline, acidic, and neutral pHs. The results showed that the changes were significantly affected by fermentative conditions. Under different conditions, pH exhibited changing profiles. When ES was fermented under alkaline conditions, pH decreased in a range of (10±1). At the beginning of alkaline fermentation, pH dropped significantly, at intervals of 4h, 4h, and 5h with sludge concentrations of 8665.6mg/L, 6498.8mg/L, and 4332.5mg/L, then it would become moderate. However, under acidic conditions, pH increased from 4 to 5. Finally, under neutral conditions pH exhibited a decrease then an increase throughout entire fermentation process. Further study showed short-chain fatty acids (SCFAs), ammonia nitrogen and cations contributed to pH change under various fermentation conditions. This study presents a novel strategy based on pH change to predict whether SCFAs reach their stable stage.

  3. Amino acids improve acid tolerance and internal pH maintenance in Bacillus cereus ATCC14579 strain.

    PubMed

    Senouci-Rezkallah, Khadidja; Schmitt, Philippe; Jobin, Michel P

    2011-05-01

    This study investigated the involvement of glutamate-, arginine- and lysine-dependent systems in the Acid Tolerance Response (ATR) of Bacillus cereus ATCC14579 strain. Cells were grown in a chemostat at external pH (pH(e)) 7.0 and 5.5. Population reduction after acid shock at pH 4.0 was strongly limited in cells grown at pH 5.5 (acid-adapted) compared with cells grown at pH 7.0 (unadapted), indicating that B. cereus cells grown at low pH(e) were able to induce a marked ATR. Glutamate, arginine and lysine enhanced the resistance of unadapted cells to pH 4.0 acid shock of 1-log or 2-log populations, respectively. Amino acids had no detectable effect on acid resistance in acid-adapted cells. An acid shock at pH 4.0 resulted in a marked drop in internal pH (pH(i)) in unadapted cells compared with acid-adapted cells. When acid shock was achieved in the presence of glutamate, arginine or lysine, pH(i) was maintained at higher values (6.31, 6.69 or 6.99, respectively) compared with pH(i) in the absence of amino acids (4.88). Acid-adapted cells maintained their pH(i) at around 6.4 whatever the condition. Agmatine (a competitive inhibitor of arginine decarboxylase) had a negative effect on the ability of B. cereus cells to survive and maintain their pH(i) during acid shock. Our data demonstrate that B. cereus is able to induce an ATR during growth at low pH. This adaptation depends on pH(i) homeostasis and is enhanced in the presence of glutamate, arginine and lysine. Hence evaluations of the pathogenicity of B. cereus must take into account its ability to adapt to acid stress. Copyright © 2010 Elsevier Ltd. All rights reserved.

  4. Evidence that electrostatic interactions between vesicle-associated membrane protein 2 and acidic phospholipids may modulate the fusion of transport vesicles with the plasma membrane.

    PubMed

    Williams, Dumaine; Vicôgne, Jérome; Zaitseva, Irina; McLaughlin, Stuart; Pessin, Jeffrey E

    2009-12-01

    The juxtamembrane domain of vesicle-associated membrane protein (VAMP) 2 (also known as synaptobrevin2) contains a conserved cluster of basic/hydrophobic residues that may play an important role in membrane fusion. Our measurements on peptides corresponding to this domain determine the electrostatic and hydrophobic energies by which this domain of VAMP2 could bind to the adjacent lipid bilayer in an insulin granule or other transport vesicle. Mutation of residues within the juxtamembrane domain that reduce the VAMP2 net positive charge, and thus its interaction with membranes, inhibits secretion of insulin granules in beta cells. Increasing salt concentration in permeabilized cells, which reduces electrostatic interactions, also results in an inhibition of insulin secretion. Similarly, amphipathic weak bases (e.g., sphingosine) that reverse the negative electrostatic surface potential of a bilayer reverse membrane binding of the positively charged juxtamembrane domain of a reconstituted VAMP2 protein and inhibit membrane fusion. We propose a model in which the positively charged VAMP and syntaxin juxtamembrane regions facilitate fusion by bridging the negatively charged vesicle and plasma membrane leaflets.

  5. Use of membrane vesicles as a simplified system for studying transport of auxin. Progress report

    SciTech Connect

    Goldsmith, M.H.

    1985-01-01

    The accumulation of indoleacetic acid (IAA) inside microsomal vesicles depends on the presence of a pH gradient and is reversible when the ..delta..pH is collapsed by ionosphores. Accumulation is stimulated by either napthylphthalamic acid or TIBA. The accumulation of IAA by the vesicles can be saturated. At concentrations of 1 ..mu..M or less, IAA, synthetic auxins, or auxin antagonists do not affect the pH gradient, but decrease the accumulation of /sup 3/H-IAA, and therefore compete specifically for uptake. Concentrations of 10 ..mu..M and above, uptake of either the auxins or weak acids is sufficient to overcome the buffering capacity of the solution within the vesicles. The collapse of the pH gradient by such high concentrations affects uptake of either /sup 3/H-IAA or /sup 14/C-BA to similar extents and thus is nonspecific. 3 refs.

  6. Influence of sodium chloride, pH, and lactic acid bacteria on anaerobic lactic acid utilization during fermented cucumber spoilage.

    PubMed

    Johanningsmeier, Suzanne D; Franco, Wendy; Perez-Diaz, Ilenys; McFeeters, Roger F

    2012-07-01

    Cucumbers are preserved commercially by natural fermentations in 5% to 8% sodium chloride (NaCl) brines. Occasionally, fermented cucumbers spoil after the primary fermentation is complete. This spoilage has been characterized by decreases in lactic acid and a rise in brine pH caused by microbial instability. Objectives of this study were to determine the combined effects of NaCl and pH on fermented cucumber spoilage and to determine the ability of lactic acid bacteria (LAB) spoilage isolates to initiate lactic acid degradation in fermented cucumbers. Cucumbers fermented with 0%, 2%, 4%, and 6% NaCl were blended into slurries (FCS) and adjusted to pH 3.2, 3.8, 4.3, and 5.0 prior to centrifugation, sterile-filtration, and inoculation with spoilage organisms. Organic acids and pH were measured initially and after 3 wk, 2, 6, 12, and 18 mo anaerobic incubation at 25 °C. Anaerobic lactic acid degradation occurred in FCS at pH 3.8, 4.3, and 5.0 regardless of NaCl concentration. At pH 3.2, reduced NaCl concentrations resulted in increased susceptibility to spoilage, indicating that the pH limit for lactic acid utilization in reduced NaCl fermented cucumbers is 3.2 or lower. Over 18 mo incubation, only cucumbers fermented with 6% NaCl to pH 3.2 prevented anaerobic lactic acid degradation by spoilage bacteria. Among several LAB species isolated from fermented cucumber spoilage, Lactobacillus buchneri was unique in its ability to metabolize lactic acid in FCS with concurrent increases in acetic acid and 1,2-propanediol. Therefore, L. buchneri may be one of multiple organisms that contribute to development of fermented cucumber spoilage. Microbial spoilage of fermented cucumbers during bulk storage causes economic losses for producers. Current knowledge is insufficient to predict or control these losses. This study demonstrated that in the absence of oxygen, cucumbers fermented with 6% sodium chloride to pH 3.2 were not subject to spoilage. However, lactic acid was degraded

  7. Effect of pH on fecal recovery of energy derived from volatile fatty acids.

    PubMed

    Kien, C L; Liechty, E A

    1987-01-01

    We assessed the effect of pH on volatilization of short-chain fatty acids during lyophilization. Acetic, propionic, valeric, and butyric acids were added to a fecal homogenate in amounts sufficient to raise the energy density by 18-27%. Fecal homogenate samples were either acidified (pH 2.8-3.2), alkalinized (pH 7.9-8.7), or left unchanged (4.0-4.8) prior to lyophilization and subsequent bomb calorimetry. Alkalinizing the fecal samples prevented the 20% loss of energy derived from each of these volatile fatty acids observed in samples either acidified or without pH adjustment. These data suggest that in energy balance studies involving subjects with active colonic fermentation, fecal samples should be alkalinized prior to lyophilization and bomb calorimetry.

  8. Navigational choice between reversal and curve during acidic pH avoidance behavior in Caenorhabditis elegans.

    PubMed

    Wakabayashi, Tokumitsu; Sakata, Kazumi; Togashi, Takuya; Itoi, Hiroaki; Shinohe, Sayaka; Watanabe, Miwa; Shingai, Ryuzo

    2015-11-19

    Under experimental conditions, virtually all behaviors of Caenorhabditis elegans are achieved by combinations of simple locomotion, including forward, reversal movement, turning by deep body bending, and gradual shallow turning. To study how worms regulate these locomotion in response to sensory information, acidic pH avoidance behavior was analyzed by using worm tracking system. In the acidic pH avoidance, we characterized two types of behavioral maneuvers that have similar behavioral sequences in chemotaxis and thermotaxis. A stereotypic reversal-turn-forward sequence of reversal avoidance caused an abrupt random reorientation, and a shallow gradual turn in curve avoidance caused non-random reorientation in a less acidic direction to avoid the acidic pH. Our results suggest that these two maneuvers were each triggered by a distinct threshold pH. A simulation study using the two-distinct-threshold model reproduced the avoidance behavior of the real worm, supporting the presence of the threshold. Threshold pH for both reversal and curve avoidance was altered in mutants with reduced or enhanced glutamatergic signaling from acid-sensing neurons. C. elegans employ two behavioral maneuvers, reversal (klinokinesis) and curve (klinotaxis) to avoid acidic pH. Unlike the chemotaxis in C. elegans, reversal and curve avoidances were triggered by absolute pH rather than temporal derivative of stimulus concentration in this behavior. The pH threshold is different between reversal and curve avoidance. Mutant studies suggested that the difference results from a differential amount of glutamate released from ASH and ASK chemosensory neurons.

  9. Dispersion of Vesicles Composed of Industrially Produced Alkyl (Oligo) Glucoside Using Diol-Boron Complexation.

    PubMed

    Aikawa, Tatsuo; Asano, Yuuka; Kondo, Takeshi; Yuasa, Makoto

    2016-07-01

    Alkyl (oligo)glucosides (AOG) are known to be environmentally compatible amphiphiles whose commercial applicability should be broadened. The present paper describes the preparation of molecular assemblies of industrially produced AOG, which is a mixture composed of different length of alkyl chains (C9-C12) with oligoglucoside moiety with a few (1-3) of glucose units. It was also described that regulation of the dispersibility of the molecular assemblies prepared by diol-boron complexation between the sugar moiety on AOG and boric acid in a dispersion medium. The molecular assembly of AOG was successfully formed by mixing AOG and cholesterols (CH). When using a suitable amount of CH (20-40 mol% with respect to AOG), the molecular assembly formed a vesicle structure. The dispersion ability of the resulting vesicle was dependent on both the boric acid concentration and pH of the dispersion medium. The light-scattering and ζ-potential measurements revealed that high concentrations (≥10 mM) of boric acid improved dispersibility the vesicles. In contrast, the vesicle agglomerated at low concentrations of boric acid (1-7.5 mM). In the absence of boric acid in dispersion medium, the vesicles were completely agglomerated. The optimum pH range for vesicle dispersion was found to be from neutral to basic (7.4-10.1). The (11)B NMR study revealed that borate ester formation occurred between boric acid and the diol of the sugar moiety on AOG vesicle. The present data suggest that borate ester formation that occurred on the surface of the vesicle provided negative charge to the vesicles, contributing to their dispersion via repulsive forces.

  10. Release of 5-Aminosalicylic Acid (5-ASA) from Mesalamine Formulations at Various pH Levels.

    PubMed

    Abinusawa, Adeyinka; Tenjarla, Srini

    2015-05-01

    Oral formulations of 5-aminosalicylic acid (5-ASA) for treatment of ulcerative colitis have been developed to minimize absorption prior to the drug reaching the colon. In this study, we investigate the release of 5-ASA from available oral mesalamine formulations in physiologically relevant pH conditions. Release of 5-ASA from 6 mesalamine formulations (APRISO®, Salix Pharmaceuticals, Inc., USA; ASACOL® MR, Procter & Gamble Pharmaceuticals UK Ltd.; ASACOL® HD, Procter & Gamble Pharmaceuticals, USA; MEZAVANT XL®, Shire US Inc.; PENTASA®, Ferring Pharmaceuticals, Ltd., UK; SALOFALK®, Dr. Falk Pharma UK Ltd.) was evaluated using United States Pharmacopeia apparatus I and II at pH values of 1.0 (2 h), 6.0 (1 h), and 6.8 (8 h). Dissolution profiles were determined for each formulation, respectively. Of the tested formulations, only the PENTASA formulation demonstrated release of 5-ASA at pH 1.0 (48%), with 56% cumulative release after exposure to pH 6.0 and 92% 5-ASA release after 6-8 h at pH 6.8. No other mesalamine formulation showed >1% drug release at pH 1.0. The APRISO formulation revealed 36% 5-ASA release at pH 6.0, with 100% release after 3 h at pH 6.8. The SALOFALK formulation revealed 11% 5-ASA release at pH 6.0, with 100% release after 1 h at pH 6.8. No 5-ASA was released by the ASACOL MR, ASACOL HD, and MEZAVANT XL formulations at pH 6.0. At pH 6.8, the ASACOL MR and ASACOL HD formulations exhibited complete release of 5-ASA after 4 and 2 h, respectively, and the MEZAVANT XL formulation demonstrated complete 5-ASA release over 6-7 h. 5-Aminosalicylic acid release profiles were variable among various commercially available formulations. Shire Development LLC.

  11. Interaction forces and membrane charge tunability: Oleic acid containing membranes in different pH conditions.

    PubMed

    Kurniawan, James; Suga, Keishi; Kuhl, Tonya L

    2017-02-01

    Oleic acid is known to interact with saturated lipid molecules and increase the fluidity of gel phase lipid membranes. In this work, the thermodynamic properties of mixed monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and oleic acid at the air-water interface were determined using Langmuir isotherms. The isotherm study revealed an attractive interaction between oleic acid and DPPC. The incorporation of oleic acid also monotonically decreased the elastic modulus of the monolayer indicative of higher fluidity with increasing oleic acid content. Using the surface force apparatus, intermembrane force-distance profiles were obtained for substrate supported DPPC membranes containing 30mol% oleic acid at pH5.8 and 7.4. Three different preparation conditions resulted in distinct force profiles. Membranes prepared in pH5.8 subphase had a low number of nanoscopic defects ≤1% and an adhesion magnitude of ~0.6mN/m. A slightly higher defect density of 1-4% was found for membranes prepared in a physiological pH7.4 subphase. The presence of the exposed hydrophobic moieties resulted in a higher adhesion magnitude of 2.9mN/m. Importantly, at pH7.4, some oleic acid deprotonates resulting in a long-range electrostatic repulsion. Even though oleic acid increased the DPPC bilayer fluidity and the number of defects, no membrane restructuring was observed indicating that the system maintained a stable configuration.

  12. Insulin Fibrillization at Acidic and Physiological pH Values is Controlled by Different Molecular Mechanisms.

    PubMed

    Noormägi, Andra; Valmsen, Karin; Tõugu, Vello; Palumaa, Peep

    2015-12-01

    Formation of amyloid-like fibrils by insulin was studied at different insulin concentrations, pH and temperatures. At low pH (pH 2.5) the insulin fibrillization occurred only at high ([10 lM) peptide concentrations, whereas at physiological pH values the fibril formation is inhibited at higher insulin concentrations. The enthalpy of activation Ea of the fibril growth at pH 2.5 equals to 33 kJ/mol, which is considerably lower than 84 kJ/mol at physiological pH. The fibrillization rate of insulin decreases with increasing pH at high, 250 lM concentration, which was opposite to the pH effect observed in 2.5 lM insulin solutions. The latter effect indicates that protonation of histidine residues seems to be important for the fibrillization of monomeric insulin, whereas the pH effect at high concentration may result from off-pathway oligomerization propensity. Together, the different effect of environmental factors on the insulin fibrillization suggest that the reaction rate is controlled by different molecular events in acidic conditions and at physiological pH values.

  13. Penetration and fusion of phospholipid vesicles by lysozyme

    SciTech Connect

    Kim, J.; Kim, H.

    1989-10-01

    The lysozyme-induced fusion of phosphatidylserine/phosphatidylethanolamine vesicles as studied at a wide range of pH is found to correlate well with the binding of this protein to the vesicles. An identical 6000 molecular weight segment of lysozyme at the N-terminal region is found to be protected from tryptic digestion when initially incubated with vesicles at several pH values. Only this segment is labeled by dansyl chloride, which is partitioned into the bilayer. These results suggest the penetration of one segment of lysozyme into the bilayer. Photoactivated labeling of the membrane-penetrating segment of lysozyme with 3-(trifluoromethyl)-3-(({sup 125}I)iodophenyl)diazirine (({sup 125}I)TID) and subsequent identification of the labeled residues by Edman degradation and gamma-ray counting indicate that four amino acids from the N-terminal are located outside the hydrophobic core of the bilayer. Although treatment of the membrane-embedded segment with aminopeptidase failed to cleave any amino acids from the N-terminal, it appears that a loop of lysozyme segment near the N-terminal penetrates into the bilayer at acidic pH. A helical wheel diagram shows that the labeling is done mainly on one surface of the alpha-helix. The penetration kinetics as studied by time-dependent ({sup 125}I)TID labeling coincide with the fusion kinetics, strongly suggesting that the penetration of the lysozyme segment into the vesicles is the cause of the fusion.

  14. Can acid (pH) refluxes predict multichannel intraluminal impedance refluxes? A correlation study.

    PubMed

    Misra, Sudipta

    2010-04-01

    The present study aims to explore if and when acid (pH) refluxes can predict refluxes detected by multichannel intraluminal impedance (MII) studies. This correlation may indicate whether pH probe-only and MII-pH studies are interchangeable. Prospective observational cross sectional study of symptomatic children (below 18 years) who had MII-pH studies done for gastroesophageal reflux. Clinical data were extracted from patient records. Non-parametric tests, Pearson's rho and receiver operating characteristic (ROC) curves were used for data analysis. A total of 153 children were included in the study and 62% were on acid suppression. Indices for acid and MII refluxes correlated with each other only in those without acid suppression. This correlation was lost in children on acid suppression. There was no statistically significant difference in acid or MII reflux indices in children with or without acid suppression. Like acid reflux, indices for MII refluxes had good correlation with each other irrespective of acid suppression. Liquid and mixed MII refluxes showed excellent correlation with respective types of proximally migrating refluxes. The values for MII reflux indices derived from our patient population were in broad agreement with available pediatric and adult data. A pH probe-only study in patients without acid suppression may reflect both acid and volume (MII) reflux activities adequately and can substitute for MII-pH study. The observed excellent correlation between acid and MII refluxes with proximal migration may justify using pH probe-only studies for extra esophageal symptoms in patients without acid suppression.

  15. Sensitivity of acid-adapted and acid-shocked Shigella flexneri to reduced pH achieved with acetic, lactic, and propionic acids.

    PubMed

    Tetteh, G L; Beuchat, L R

    2001-07-01

    Survival and growth characteristics of unadapted, acid-adapted, and acid-shocked Shigella flexneri 2a cells in acidified (pH 3.5 to 5.5) tryptic soy broth with 0.25% glucose (TSB) and tryptic soy agar (TSA) were determined. S. flexneri was grown at 37 degrees C for 18 h in tryptic soy broth without glucose (TSBNG) (unadapted) and TSBNG supplemented with 1% glucose (TSBG) (acid-adapted). Cells grown in TSBNG were acid shocked by adjusting 16-h cultures to pH 5.05 +/- 0.05 with lactic acid. Cells were then inoculated into TSB acidified with acetic, lactic, or propionic acids to pH 5.5, 4.5, or 3.5 and incubated at 37 degrees C for 6 h. The order of lethality at a given pH was lactic acid < acetic acid < propionic acid. Significantly (P < or = 0.05) higher numbers of acid-adapted cells, compared to acid-shocked and unadapted cells, were recovered from TSB acidified (pH 3.5) with lactic or acetic acids. None of the cells survived a 30-min exposure in TSB acidified with propionic acid to pH 3.5. When the three cell types were plated on TSA acidified with lactic, acetic, or propionic acids at pH < or = 4.5, < or = 5.5, and < or = 5.5, respectively, visible colonies were not detected. Viable unadapted, acid-adapted, and acid-shocked cells were, however, recovered from TSA acidified with all three acids at pH > or = 4.5. Acid-adapted and, to a lesser extent, acid-shocked cells survived at lower pH than did unadapted cells, indicating that prior exposure to mild acidic environment results in increased acid resistance. Survival of S. flexneri at a given pH was influenced by the type of acidulant used, a response characteristic exhibited by other gram-negative enteric pathogens.

  16. The pH ruler: a Java applet for developing interactive exercises on acids and bases.

    PubMed

    Barrette-Ng, Isabelle H

    2011-07-01

    In introductory biochemistry courses, it is often a struggle to teach the basic concepts of acid-base chemistry in a manner that is relevant to biological systems. To help students gain a more intuitive and visual understanding of abstract acid-base concepts, a simple graphical construct called the pH ruler Java applet was developed. The applet allows students to visualize the abundance of different protonation states of diprotic and triprotic amino acids at different pH values. Using the applet, the student can drag a widget on a slider bar to change the pH and observe in real time changes in the abundance of different ionization states of this amino acid. This tool provides a means for developing more complex inquiry-based, active-learning exercises to teach more advanced topics of biochemistry, such as protein purification, protein structure and enzyme mechanism.

  17. Numerical interdependence in pH, acidity and alkalinity of a polluted river water.

    PubMed

    Shashi; Singh, J; Dwivedi, Anil K

    2009-09-01

    Acidity and alkalinity are independent parameters and they directly or indirectly regulate the pH of a medium. These are the prime factors which control the nature of the reactions in a medium. The present study was designed to explore the numerical interdependence of the three parameters and also, to develop the regression models. For physico-chemical analysis of the selected parameters, water quality of a polluted tropical river was analysed fortnightly, for complete one year. Correlation coefficients between pH-acidity and acidity-alkalinity were negative while that between pH and alkalinity were positive. The value calculated by the developed multiple regression models, explain pH, acidity and alkalinity up to 57, 65 and 79% respectively, suggesting their utility and relevance.

  18. Photoproduction of glyoxylic acid in model wine: Impact of sulfur dioxide, caffeic acid, pH and temperature.

    PubMed

    Grant-Preece, Paris; Schmidtke, Leigh M; Barril, Celia; Clark, Andrew C

    2017-01-15

    Glyoxylic acid is a tartaric acid degradation product formed in model wine solutions containing iron and its production is greatly increased by exposure to UV-visible light. In this study, the combined effect of sulfur dioxide, caffeic acid, pH and temperature on the light-induced (⩾300nm) production of glyoxylic acid in model wine containing tartaric acid and iron was investigated using a Box-Behnken experimental design and response surface methodology (RSM). Glyoxylic acid produced in the irradiated model wine was present in free and hydrogen sulfite adduct forms and the measured total, free and percentage free glyoxylic acid values were modeled using RSM. Sulfur dioxide significantly decreased the total amount of glyoxylic acid produced, but could not prevent its production, while caffeic acid showed no significant impact. The interaction between pH and temperature was significant, with low pH values and low temperatures giving rise to higher levels of total glyoxylic acid. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Effect of pH on conjugated linoleic acid (CLA) formation of linolenic acid biohydrogenation by ruminal microorganisms.

    PubMed

    Lee, Yongjae

    2013-08-01

    Conventional beliefs surrounding the linolenic acid (LNA; cis-9 cis-12 cis-15 C18:3) biohydrogenation (BH) pathway propose that it converts to stearic acid (SA) without the formation of conjugated linoleic acid (CLA) as intermediate isomers. However, an advanced study (Lee and Jenkins, 2011) verified that LNA BH yields multiple CLAs. This study utilized the stable isotope tracer to investigate the BH intermediates of (13)C-LNA with different pH conditions (5.5 and 6.5). The (13)C enrichment was calculated as a (13)C/(12)C ratio of labeled minus unlabeled. After 24 h, eight CLA isomers were significantly enriched on both pH treatment, this result verifies that these CLAs originated from (13)C-LNA BH which supports the results of Lee and Jenkins (2011). The enrichment of cis-cis double bond CLAs (cis-9 cis-11 and cis-10 cis-12 CLA) were significantly higher at low pH conditions. Furthermore, the concentration of cis-10 cis-12 CLA at low pH was four times higher than at high pH conditions after a 3 h incubation. These differences support the LNA BH pathways partial switch under different pH conditions, with a strong influence on the cis-cis CLA at low pH. Several mono-, di-, and tri-enoic fatty acid isomers were enriched during 24 h of incubation, but the enrichment was decreased or restricted at low pH treatment. Based on these results, it is proposed that low pH conditions may cause a changed or limited capacity of the isomerization and reduction steps in BH.

  20. Acidic Food pH Increases Palatability and Consumption and Extends Drosophila Lifespan12

    PubMed Central

    Deshpande, Sonali A; Yamada, Ryuichi; Mak, Christine M; Hunter, Brooke; Obando, Alina Soto; Hoxha, Sany; Ja, William W

    2015-01-01

    Background: Despite the prevalent use of Drosophila as a model in studies of nutrition, the effects of fundamental food properties, such as pH, on animal health and behavior are not well known. Objectives: We examined the effect of food pH on adult Drosophila lifespan, feeding behavior, and microbiota composition and tested the hypothesis that pH-mediated changes in palatability and total consumption are required for modulating longevity. Methods: We measured the effect of buffered food (pH 5, 7, or 9) on male gustatory responses (proboscis extension), total food intake, and male and female lifespan. The effect of food pH on germfree male lifespan was also assessed. Changes in fly-associated microbial composition as a result of food pH were determined by 16S ribosomal RNA gene sequencing. Male gustatory responses, total consumption, and male and female longevity were additionally measured in the taste-defective Pox neuro (Poxn) mutant and its transgenic rescue control. Results: An acidic diet increased Drosophila gustatory responses (40–230%) and food intake (5–50%) and extended survival (10–160% longer median lifespan) compared with flies on either neutral or alkaline pH food. Alkaline food pH shifted the composition of fly-associated bacteria and resulted in greater lifespan extension (260% longer median survival) after microbes were eliminated compared with flies on an acidic (50%) or neutral (130%) diet. However, germfree flies lived longer on an acidic diet (5–20% longer median lifespan) compared with those on either neutral or alkaline pH food. Gustatory responses, total consumption, and longevity were unaffected by food pH in Poxn mutant flies. Conclusions: Food pH can directly influence palatability and feeding behavior and affect parameters such as microbial growth to ultimately affect Drosophila lifespan. Fundamental food properties altered by dietary or drug interventions may therefore contribute to changes in animal physiology, metabolism, and

  1. Acidic Food pH Increases Palatability and Consumption and Extends Drosophila Lifespan.

    PubMed

    Deshpande, Sonali A; Yamada, Ryuichi; Mak, Christine M; Hunter, Brooke; Soto Obando, Alina; Hoxha, Sany; Ja, William W

    2015-12-01

    Despite the prevalent use of Drosophila as a model in studies of nutrition, the effects of fundamental food properties, such as pH, on animal health and behavior are not well known. We examined the effect of food pH on adult Drosophila lifespan, feeding behavior, and microbiota composition and tested the hypothesis that pH-mediated changes in palatability and total consumption are required for modulating longevity. We measured the effect of buffered food (pH 5, 7, or 9) on male gustatory responses (proboscis extension), total food intake, and male and female lifespan. The effect of food pH on germfree male lifespan was also assessed. Changes in fly-associated microbial composition as a result of food pH were determined by 16S ribosomal RNA gene sequencing. Male gustatory responses, total consumption, and male and female longevity were additionally measured in the taste-defective Pox neuro (Poxn) mutant and its transgenic rescue control. An acidic diet increased Drosophila gustatory responses (40-230%) and food intake (5-50%) and extended survival (10-160% longer median lifespan) compared with flies on either neutral or alkaline pH food. Alkaline food pH shifted the composition of fly-associated bacteria and resulted in greater lifespan extension (260% longer median survival) after microbes were eliminated compared with flies on an acidic (50%) or neutral (130%) diet. However, germfree flies lived longer on an acidic diet (5-20% longer median lifespan) compared with those on either neutral or alkaline pH food. Gustatory responses, total consumption, and longevity were unaffected by food pH in Poxn mutant flies. Food pH can directly influence palatability and feeding behavior and affect parameters such as microbial growth to ultimately affect Drosophila lifespan. Fundamental food properties altered by dietary or drug interventions may therefore contribute to changes in animal physiology, metabolism, and survival. © 2015 American Society for Nutrition.

  2. pH of Aerosols in a Polluted Atmosphere: Source Contributions to Highly Acidic Aerosol.

    PubMed

    Shi, Guoliang; Xu, Jiao; Peng, Xing; Xiao, Zhimei; Chen, Kui; Tian, Yingze; Guan, Xinbei; Feng, Yinchang; Yu, Haofei; Nenes, Athanasios; Russell, Armistead G

    2017-04-18

    Acidity (pH) plays a key role in the physical and chemical behavior of PM2.5. However, understanding of how specific PM sources impact aerosol pH is rarely considered. Performing source apportionment of PM2.5 allows a unique link of sources pH of aerosol from the polluted city. Hourly water-soluble (WS) ions of PM2.5 were measured online from December 25th, 2014 to June 19th, 2015 in a northern city in China. Five sources were resolved including secondary nitrate (41%), secondary sulfate (26%), coal combustion (14%), mineral dust (11%), and vehicle exhaust (9%). The influence of source contributions to pH was estimated by ISORROPIA-II. The lowest aerosol pH levels were found at low WS-ion levels and then increased with increasing total ion levels, until high ion levels occur, at which point the aerosol becomes more acidic as both sulfate and nitrate increase. Ammonium levels increased nearly linearly with sulfate and nitrate until approximately 20 μg m(-3), supporting that the ammonium in the aerosol was more limited by thermodynamics than source limitations, and aerosol pH responded more to the contributions of sources such as dust than levels of sulfate. Commonly used pH indicator ratios were not indicative of the pH estimated using the thermodynamic model.

  3. Properties of acid whey as a function of pH and temperature.

    PubMed

    Chandrapala, Jayani; Duke, Mikel C; Gray, Stephen R; Zisu, Bogdan; Weeks, Mike; Palmer, Martin; Vasiljevic, Todor

    2015-07-01

    Compositional differences of acid whey (AW) in comparison with other whey types limit its processability and application of conventional membrane processing. Hence, the present study aimed to identify chemical and physical properties of AW solutions as a function of pH (3 to 10.5) at 4 different temperatures (15, 25, 40, or 90°C) to propose appropriate membrane-processing conditions for efficient use of AW streams. The concentration of minerals, mainly calcium and phosphate, and proteins in centrifuged supernatants was significantly lowered with increase in either pH or temperature. Lactic acid content decreased with pH decline and rose at higher temperatures. Calcium appeared to form complexes with phosphates and lactates mainly, which in turn may have induced molecular attractions with the proteins. An increase in pH led to more soluble protein aggregates with large particle sizes. Surface hydrophobicity of these particles increased significantly with temperature up to 40°C and decreased with further heating to 90°C. Surface charge was clearly pH dependent. High lactic acid concentrations appeared to hinder protein aggregation by hydrophobic interactions and may also indirectly influence protein denaturation. Processing conditions such as pH and temperature need to be optimized to manipulate composition, state, and surface characteristics of components of AW systems to achieve an efficient separation and concentration of lactic acid and lactose.

  4. Monascus ruber as cell factory for lactic acid production at low pH.

    PubMed

    Weusthuis, Ruud A; Mars, Astrid E; Springer, Jan; Wolbert, Emil Jh; van der Wal, Hetty; de Vrije, Truus G; Levisson, Mark; Leprince, Audrey; Houweling-Tan, G Bwee; Pha Moers, Antoine; Hendriks, Sjon Na; Mendes, Odette; Griekspoor, Yvonne; Werten, Marc Wt; Schaap, Peter J; van der Oost, John; Eggink, Gerrit

    2017-07-01

    A Monascus ruber strain was isolated that was able to grow on mineral medium at high sugar concentrations and 175g/l lactic acid at pH 2.8. Its genome and transcriptomes were sequenced and annotated. Genes encoding lactate dehydrogenase (LDH) were introduced to accomplish lactic acid production and two genes encoding pyruvate decarboxylase (PDC) were knocked out to subdue ethanol formation. The strain preferred lactic acid to glucose as carbon source, which hampered glucose consumption and therefore also lactic acid production. Lactic acid consumption was stopped by knocking out 4 cytochrome-dependent LDH (CLDH) genes, and evolutionary engineering was used to increase the glucose consumption rate. Application of this strain in a fed-batch fermentation resulted in a maximum lactic acid titer of 190g/l at pH 3.8 and 129g/l at pH 2.8, respectively 1.7 and 2.2 times higher than reported in literature before. Yield and productivity were on par with the best strains described in literature for lactic acid production at low pH. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  5. Microbial degradation of isosaccharinic acid at high pH

    PubMed Central

    Bassil, Naji M; Bryan, Nicholas; Lloyd, Jonathan R

    2015-01-01

    Intermediate-level radioactive waste (ILW), which dominates the radioactive waste inventory in the United Kingdom on a volumetric basis, is proposed to be disposed of via a multibarrier deep geological disposal facility (GDF). ILW is a heterogeneous wasteform that contains substantial amounts of cellulosic material encased in concrete. Upon resaturation of the facility with groundwater, alkali conditions will dominate and will lead to the chemical degradation of cellulose, producing a substantial amount of organic co-contaminants, particularly isosaccharinic acid (ISA). ISA can form soluble complexes with radionuclides, thereby mobilising them and posing a potential threat to the surrounding environment or ‘far field'. Alkaliphilic microorganisms sampled from a legacy lime working site, which is an analogue for an ILW-GDF, were able to degrade ISA and couple this degradation to the reduction of electron acceptors that will dominate as the GDF progresses from an aerobic ‘open phase' through nitrate- and Fe(III)-reducing conditions post closure. Furthermore, pyrosequencing analyses showed that bacterial diversity declined as the reduction potential of the electron acceptor decreased and that more specialised organisms dominated under anaerobic conditions. These results imply that the microbial attenuation of ISA and comparable organic complexants, initially present or formed in situ, may play a role in reducing the mobility of radionuclides from an ILW-GDF, facilitating the reduction of undue pessimism in the long-term performance assessment of such facilities. PMID:25062127

  6. Role of acid pH and deficient efflux of pyrazinoic acid in unique susceptibility of Mycobacterium tuberculosis to pyrazinamide.

    PubMed

    Zhang, Y; Scorpio, A; Nikaido, H; Sun, Z

    1999-04-01

    Pyrazinamide (PZA) is an important antituberculosis drug. Unlike most antibacterial agents, PZA, despite its remarkable in vivo activity, has no activity against Mycobacterium tuberculosis in vitro except at an acidic pH. M. tuberculosis is uniquely susceptible to PZA, but other mycobacteria as well as nonmycobacteria are intrinsically resistant. The role of acidic pH in PZA action and the basis for the unique PZA susceptibility of M. tuberculosis are unknown. We found that in M. tuberculosis, acidic pH enhanced the intracellular accumulation of pyrazinoic acid (POA), the active derivative of PZA, after conversion of PZA by pyrazinamidase. In contrast, at neutral or alkaline pH, POA was mainly found outside M. tuberculosis cells. PZA-resistant M. tuberculosis complex organisms did not convert PZA into POA. Unlike M. tuberculosis, intrinsically PZA-resistant M. smegmatis converted PZA into POA, but it did not accumulate POA even at an acidic pH, due to a very active POA efflux mechanism. We propose that a deficient POA efflux mechanism underlies the unique susceptibility of M. tuberculosis to PZA and that the natural PZA resistance of M. smegmatis is due to a highly active efflux pump. These findings may have implications with regard to the design of new antimycobacterial drugs.

  7. Natively unfolded human prothymosin alpha adopts partially folded collapsed conformation at acidic pH.

    PubMed

    Uversky, V N; Gillespie, J R; Millett, I S; Khodyakova, A V; Vasiliev, A M; Chernovskaya, T V; Vasilenko, R N; Kozlovskaya, G D; Dolgikh, D A; Fink, A L; Doniach, S; Abramov, V M

    1999-11-09

    Prothymosin alpha has previously been shown to be unfolded at neutral pH, thus belonging to a growing family of "natively unfolded" proteins. The structural properties and conformational stability of recombinant human prothymosin alpha were characterized at neutral and acidic pH by gel filtration, SAXS, circular dichroism, ANS fluorescence, (1)H NMR, and resistance to urea-induced unfolding. Interestingly, prothymosin alpha underwent a cooperative transition from the unfolded state into a partially folded conformation on lowering the pH. This conformation of prothymosin alpha is a compact denatured state, with structural properties different from those of the molten globule. The formation of alpha-helical structure by the glutamic acid-rich elements of the protein accompanied by the partial hydrophobic collapse is expected at lower pH due to the neutralization of the negatively charged residues. It is possible that such conformational changes may be associated with the protein function.

  8. Effects of pH adjustment and sodium ions on sour taste intensity of organic acids.

    PubMed

    Neta, E R D; Johanningsmeier, S D; Drake, M A; McFeeters, R F

    2009-01-01

    Protonated organic acid species have been shown to be the primary stimuli responsible for sour taste of organic acids. However, we have observed that sour taste may be modulated when the pH of acid solutions is raised using sodium hydroxide. Objectives were to evaluate the effect of pH adjustment on sour taste of equimolar protonated organic acid solutions and to investigate the potential roles of organic anions and sodium ions on sour taste perception. Despite equal concentrations of protonated acid species, sour taste intensity decreased significantly with increased pH for acetic, lactic, malic, and citric acids (P < 0.05). Total organic anion concentration did not explain the suppression of sour taste in solutions containing a blend of 3 organic acids with constant concentration of protonated organic acid species and hydrogen ions and variable organic anion concentrations (R(2)= 0.480, P = 0.12). Sour taste suppression in these solutions seemed to be more closely related to sodium ions added in the form of NaOH (R(2)= 0.861, P = 0.007). Addition of 20 mM NaCl to acid solutions resulted in significant suppression of sour taste (P = 0.016). However, sour taste did not decrease with further addition of NaCl up to 80 mM. Presence of sodium ions was clearly shown to decrease sour taste of organic acid solutions. Nonetheless, suppression of sour taste in pH adjusted single acid solutions was greater than what would be expected based on the sodium ion concentration alone, indicating an additional suppression mechanism may be involved.

  9. Effect of benzoic acid on glycolytic metabolite levels and intracellular pH in Saccharomyces cerevisiae.

    PubMed Central

    Warth, A D

    1991-01-01

    Low concentrations of benzoic acid stimulated fermentation rates in Saccharomyces cerevisiae. At concentrations near the maximum permitting growth, there was inhibition of fermentation, lowered ATP and intracellular pH, and relatively greater accumulation of benzoate. Changes in the levels of glycolytic intermediates suggested that fermentation was inhibited as a result of high ATP usage rather than of lowered intracellular pH. Specific inhibition of phosphofructokinase or of several other glycolytic enzymes was not observed. PMID:1785917

  10. Initial pH of medium affects organic acids production but do not affect phosphate solubilization.

    PubMed

    Marra, Leandro M; de Oliveira-Longatti, Silvia M; Soares, Cláudio R F S; de Lima, José M; Olivares, Fabio L; Moreira, Fatima M S

    2015-06-01

    The pH of the culture medium directly influences the growth of microorganisms and the chemical processes that they perform. The aim of this study was to assess the influence of the initial pH of the culture medium on the production of 11 low-molecular-weight organic acids and on the solubilization of calcium phosphate by bacteria in growth medium (NBRIP). The following strains isolated from cowpea nodules were studied: UFLA03-08 (Rhizobium tropici), UFLA03-09 (Acinetobacter sp.), UFLA03-10 (Paenibacillus kribbensis), UFLA03-106 (Paenibacillus kribbensis) and UFLA03-116 (Paenibacillus sp.). The strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 solubilized Ca3(PO4)2 in liquid medium regardless of the initial pH, although without a significant difference between the treatments. The production of organic acids by these strains was assessed for all of the initial pH values investigated, and differences between the treatments were observed. Strains UFLA03-09 and UFLA03-10 produced the same acids at different initial pH values in the culture medium. There was no correlation between phosphorus solubilized from Ca3(PO4)2 in NBRIP liquid medium and the concentration of total organic acids at the different initial pH values. Therefore, the initial pH of the culture medium influences the production of organic acids by the strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 but it does not affect calcium phosphate solubilization.

  11. Initial pH of medium affects organic acids production but do not affect phosphate solubilization

    PubMed Central

    Marra, Leandro M.; de Oliveira-Longatti, Silvia M.; Soares, Cláudio R.F.S.; de Lima, José M.; Olivares, Fabio L.; Moreira, Fatima M.S.

    2015-01-01

    The pH of the culture medium directly influences the growth of microorganisms and the chemical processes that they perform. The aim of this study was to assess the influence of the initial pH of the culture medium on the production of 11 low-molecular-weight organic acids and on the solubilization of calcium phosphate by bacteria in growth medium (NBRIP). The following strains isolated from cowpea nodules were studied: UFLA03-08 (Rhizobium tropici), UFLA03-09 (Acinetobacter sp.), UFLA03-10 (Paenibacillus kribbensis), UFLA03-106 (Paenibacillus kribbensis) and UFLA03-116 (Paenibacillus sp.). The strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 solubilized Ca3(PO4)2 in liquid medium regardless of the initial pH, although without a significant difference between the treatments. The production of organic acids by these strains was assessed for all of the initial pH values investigated, and differences between the treatments were observed. Strains UFLA03-09 and UFLA03-10 produced the same acids at different initial pH values in the culture medium. There was no correlation between phosphorus solubilized from Ca3(PO4)2 in NBRIP liquid medium and the concentration of total organic acids at the different initial pH values. Therefore, the initial pH of the culture medium influences the production of organic acids by the strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 but it does not affect calcium phosphate solubilization. PMID:26273251

  12. Acidic duodenal pH alters gene expression in the cystic fibrosis mouse pancreas.

    PubMed

    Kaur, Simran; Norkina, Oxana; Ziemer, Donna; Samuelson, Linda C; De Lisle, Robert C

    2004-08-01

    The duodenum is abnormally acidic in cystic fibrosis (CF) due to decreased bicarbonate ion secretion that is dependent on the CF gene product CFTR. In the CFTR null mouse, the acidic duodenum results in increased signaling from the intestine to the exocrine pancreas in an attempt to stimulate pancreatic bicarbonate ion secretion. Excess stimulation is proposed to add to the stress/inflammation of the pancreas in CF. DNA microarray analysis of the CF mouse revealed altered pancreatic gene expression characteristic of stress/inflammation. When the duodenal pH was corrected genetically (crossing CFTR null with gastrin null mice) or pharmacologically (use of the proton pump inhibitor omeprazole), expression levels of genes measured by quantitative RT-PCR were significantly normalized. It is concluded that the acidic duodenal pH in CF contributes to the stress on the exocrine pancreas and that normalizing duodenal pH reduces this stress.

  13. Kinetics of salivary pH after acidic beverage intake by patients undergoing orthodontic treatment.

    PubMed

    Turssi, Cecilia P; Silva, Carolina S; Bridi, Enrico C; Amaral, Flavia Lb; Franca, Fabiana Mg; Basting, Roberta T

    2015-01-01

    The saliva of patients undergoing orthodontic treatment with fixed appliances can potentially present a delay in the diluting, clearing, and buffering of dietary acids due to an increased number of retention areas. The aim of this clinical trial was to compare salivary pH kinetics of patients with and without orthodontic treatment, following the intake of an acidic beverage. Twenty participants undergoing orthodontic treatment and 20 control counterparts had their saliva assessed for flow rate, pH, and buffering capacity. There was no significant difference between salivary parameters in participants with or without an orthodontic appliance. Salivary pH recovery following acidic beverage intake was slower in the orthodontic subjects compared to controls. Patients with fixed orthodontic appliances, therefore, seem to be at higher risk of dental erosion, suggesting that dietary advice and preventive care need to be implemented during orthodontic treatment.

  14. Differential gene expression in tomato fruit and Colletotrichum gloeosporioides during colonization of the RNAi-SlPH tomato line with reduced fruit acidity and higher pH.

    PubMed

    Barad, Shiri; Sela, Noa; Dubey, Amit K; Kumar, Dilip; Luria, Neta; Ment, Dana; Cohen, Shahar; Schaffer, Arthur A; Prusky, Dov

    2017-08-04

    The destructive phytopathogen Colletotrichum gloeosporioides causes anthracnose disease in fruit. During host colonization, it secretes ammonia, which modulates environmental pH and regulates gene expression, contributing to pathogenicity. However, the effect of host pH environment on pathogen colonization has never been evaluated. Development of an isogenic tomato line with reduced expression of the gene for acidity, SlPH (Solyc10g074790.1.1), enabled this analysis. Total RNA from C. gloeosporioides colonizing wild-type (WT) and RNAi-SlPH tomato lines was sequenced and gene-expression patterns were compared. C. gloeosporioides inoculation of the RNAi-SlPH line with pH 5.96 compared to the WT line with pH 4.2 showed 30% higher colonization and reduced ammonia accumulation. Large-scale comparative transcriptome analysis of the colonized RNAi-SlPH and WT lines revealed their different mechanisms of colonization-pattern activation: whereas the WT tomato upregulated 13-LOX (lipoxygenase), jasmonic acid and glutamate biosynthesis pathways, it downregulated processes related to chlorogenic acid biosynthesis II, phenylpropanoid biosynthesis and hydroxycinnamic acid tyramine amide biosynthesis; the RNAi-SlPH line upregulated UDP-D-galacturonate biosynthesis I and free phenylpropanoid acid biosynthesis, but mainly downregulated pathways related to sugar metabolism, such as the glyoxylate cycle and L-arabinose degradation II. Comparison of C. gloeosporioides gene expression during colonization of the WT and RNAi-SlPH lines showed that the fungus upregulates ammonia and nitrogen transport and the gamma-aminobutyric acid metabolic process during colonization of the WT, while on the RNAi-SlPH tomato, it mainly upregulates the nitrate metabolic process. Modulation of tomato acidity and pH had significant phenotypic effects on C. gloeosporioides development. The fungus showed increased colonization on the neutral RNAi-SlPH fruit, and limited colonization on the WT acidic fruit

  15. Self-assembly of ketals of arjunolic acid into vesicles and fibers yielding gel-like dispersions.

    PubMed

    Bag, Braja G; Majumdar, Rakhi; Dinda, Shaishab K; Dey, Partha P; Maity, Gopal C; Mallia, V Ajay; Weiss, Richard G

    2013-02-12

    Ten aliphatic and aromatic ketals of arjunolic acid, a renewable, nanosized triterpenic acid which is obtainable from Terminalia arjuna, have been synthesized upon condensation with aldehydes. Self-assembly properties of the ketals have been studied in a wide range of organic liquids. With the exception of the p-nitrobenzylidene derivative, low concentrations of the ketals self-assemble and form gel-like dispersions in many of the organic liquids examined. The morphologies of the assemblies, studied at different distance scales by optical, electron, and atomic-force microscopies, consisted of fibrillar networks and vesicles which were able to entrap 5(6)-carboxyfluorescein as a guest molecule. X-ray diffractograms indicate that the fibrillar objects are crystalline. A charge-transfer complex was formed from a 1:1 mixture of ketal derivatives with electron-donating and electron-accepting groups, and the 9-anthrylidene derivative in its fibrillar network dimerized upon irradiation. Results demonstrate that subtle changes in the ketal structures can lead to very different aggregation pathways.

  16. Influence of Acidic pH on Hydrogen and Acetate Production by an Electrosynthetic Microbiome

    PubMed Central

    LaBelle, Edward V.; Marshall, Christopher W.; Gilbert, Jack A.; May, Harold D.

    2014-01-01

    Production of hydrogen and organic compounds by an electrosynthetic microbiome using electrodes and carbon dioxide as sole electron donor and carbon source, respectively, was examined after exposure to acidic pH (∼5). Hydrogen production by biocathodes poised at −600 mV vs. SHE increased>100-fold and acetate production ceased at acidic pH, but ∼5–15 mM (catholyte volume)/day acetate and>1,000 mM/day hydrogen were attained at pH ∼6.5 following repeated exposure to acidic pH. Cyclic voltammetry revealed a 250 mV decrease in hydrogen overpotential and a maximum current density of 12.2 mA/cm2 at −765 mV (0.065 mA/cm2 sterile control at −800 mV) by the Acetobacterium-dominated community. Supplying −800 mV to the microbiome after repeated exposure to acidic pH resulted in up to 2.6 kg/m3/day hydrogen (≈2.6 gallons gasoline equivalent), 0.7 kg/m3/day formate, and 3.1 kg/m3/day acetate ( = 4.7 kg CO2 captured). PMID:25333313

  17. Transcriptome Profiling of Shewanella oneidensis Gene Expressionfollowing Exposure to Acidic and Alkaline pH

    SciTech Connect

    Leaphart, Adam B.; Thompson, Dorothea K.; Huang, Katherine; Alm,Eric; Wan, Xiu-Feng; Arkin, Adam P.; Brown, Steven D.; Wu, Liyou; Yan,Tingfen; Liu, Xueduan; Wickham, Gene S.; Zhou, Jizhong

    2007-04-02

    The molecular response of Shewanella oneidensis MR-1 tovariations in extracellular pH was investigated based on genomewide geneexpression profiling. Microarray analysis revealed that cells elicitedboth general and specific transcriptome responses when challenged withenvironmental acid (pH 4) or base (pH 10) conditions over a 60-minperiod. Global responses included the differential expression of genesfunctionally linked to amino acid metabolism, transcriptional regulationand signal transduction, transport, cell membrane structure, andoxidative stress protection. Response to acid stress included theelevated expression of genes encoding glycogen biosynthetic enzymes,phosphate transporters, and the RNA polymerase sigma-38 factor (rpoS),whereas the molecular response to alkaline pH was characterized byupregulation of nhaA and nhaR, which are predicted to encode an Na+/H+antiporter and transcriptional activator, respectively, as well assulfate transport and sulfur metabolism genes. Collectively, theseresults suggest that S. oneidensis modulates multiple transporters, cellenvelope components, and pathways of amino acid consumption and centralintermediary metabolism as part of its transcriptome response to changingexternal pH conditions.

  18. Influence of acidic pH on hydrogen and acetate production by an electrosynthetic microbiome

    DOE PAGES

    LaBelle, Edward V.; Marshall, Christopher W.; Gilbert, Jack A.; ...

    2014-10-15

    Production of hydrogen and organic compounds by an electrosynthetic microbiome using electrodes and carbon dioxide as sole electron donor and carbon source, respectively, was examined after exposure to acidic pH (~5). Hydrogen production by biocathodes poised at -600 mV vs. SHE increased>100-fold and acetate production ceased at acidic pH, but ~5–15 mM (catholyte volume)/day acetate and>1,000 mM/day hydrogen were attained at pH ~6.5 following repeated exposure to acidic pH. Cyclic voltammetry revealed a 250 mV decrease in hydrogen overpotential and a maximum current density of 12.2 mA/cm2 at -765 mV (0.065 mA/cm2 sterile control at -800 mV) by the Acetobacterium-dominatedmore » community. Supplying -800 mV to the microbiome after repeated exposure to acidic pH resulted in up to 2.6 kg/m3/day hydrogen (≈2.6 gallons gasoline equivalent), 0.7 kg/m3/day formate, and 3.1 kg/m3/day acetate ( = 4.7 kg CO2 captured).« less

  19. Influence of acidic pH on hydrogen and acetate production by an electrosynthetic microbiome

    SciTech Connect

    LaBelle, Edward V.; Marshall, Christopher W.; Gilbert, Jack A.; May, Harold D.; Battista, John R.

    2014-10-15

    Production of hydrogen and organic compounds by an electrosynthetic microbiome using electrodes and carbon dioxide as sole electron donor and carbon source, respectively, was examined after exposure to acidic pH (~5). Hydrogen production by biocathodes poised at -600 mV vs. SHE increased>100-fold and acetate production ceased at acidic pH, but ~5–15 mM (catholyte volume)/day acetate and>1,000 mM/day hydrogen were attained at pH ~6.5 following repeated exposure to acidic pH. Cyclic voltammetry revealed a 250 mV decrease in hydrogen overpotential and a maximum current density of 12.2 mA/cm2 at -765 mV (0.065 mA/cm2 sterile control at -800 mV) by the Acetobacterium-dominated community. Supplying -800 mV to the microbiome after repeated exposure to acidic pH resulted in up to 2.6 kg/m3/day hydrogen (≈2.6 gallons gasoline equivalent), 0.7 kg/m3/day formate, and 3.1 kg/m3/day acetate ( = 4.7 kg CO2 captured).

  20. Influence of pH on organic acid production by Clostridium sporogenes in test tube and fermentor cultures.

    PubMed Central

    Montville, T J; Parris, N; Conway, L K

    1985-01-01

    The influence of pH on the growth parameters of and the organic acids produced by Clostridium sporogenes 3121 cultured in test tubes and fermentors at 35 degrees C was examined. Specific growth rates in the fermentor maintained at a constant pH ranged from 0.20 h-1 at pH 5.00 to 0.86 h-1 at pH 6.50. Acetic acid was the primary organic acid in supernatants of 24-h cultures; total organic acid levels were 2.0 to 22.0 mumol/ml. Supernatants from pH 5.00 and 5.50 cultures had total organic acid levels less than one-third of those found at pH 6.00 to 7.00. The specific growth rates of the test tube cultures ranged from 0.51 h-1 at pH 5.00 to 0.95 h-1 at pH 6.50. The pH of the medium did not affect the average total organic acid content (51.5 mumol/ml) but did affect the distribution of the organic acids, which included formic, acetic, propionic, butyric, 3-(p-hydroxyphenyl)propionic, and 3-phenylpropionic acids. Butyric acid levels were lower, but formic and propionic acid levels were higher, at pH 5.00 than at other pHs. PMID:4004207

  1. Life at acidic pH imposes an increased energetic cost for a eukaryotic acidophile.

    PubMed

    Messerli, Mark A; Amaral-Zettler, Linda A; Zettler, Erik; Jung, Sung-Kwon; Smith, Peter J S; Sogin, Mitchell L

    2005-07-01

    Organisms growing in acidic environments, pH<3, would be expected to possess fundamentally different molecular structures and physiological controls in comparison with similar species restricted to neutral pH. We begin to investigate this premise by determining the magnitude of the transmembrane electrochemical H+ gradient in an acidophilic Chlamydomonas sp. (ATCC PRA-125) isolated from the Rio Tinto, a heavy metal laden, acidic river (pH 1.7-2.5). This acidophile grows most rapidly at pH 2 but is capable of growth over a wide pH range (1.5-7.0), while Chlamydomonas reinhardtii is restricted to growth at pH>or=3 with optimal growth between pH 5.5 and 8.5. With the fluorescent H+ indicator, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF), we show that the acidophilic Chlamydomonas maintains an average cytosolic pH of 6.6 in culture medium at both pH 2 and pH 7 while Chlamydomonas reinhardtii maintains an average cytosolic pH of 7.1 in pH 7 culture medium. The transmembrane electric potential difference of Chlamydomonas sp., measured using intracellular electrodes at both pH 2 and 7, is close to 0 mV, a rare value for plants, animals and protists. The 40,000-fold difference in [H+] could be the result of either active or passive mechanisms. Evidence for active maintenance was detected by monitoring the rate of ATP consumption. At the peak, cells consume about 7% more ATP per second in medium at pH 2 than at pH 7. This increased rate of consumption is sufficient to account for removal of H+ entering the cytosol across a membrane with relatively high permeability to H+ (7x10(-8) cm s-1). Our results indicate that the small increase in the rate of ATP consumption can account for maintenance of the transmembrane H+ gradient without the imposition of cell surface H+ barriers.

  2. Effects of saliva on starch-thickened drinks with acidic and neutral pH.

    PubMed

    Hanson, Ben; Cox, Ben; Kaliviotis, Efstathios; Smith, Christina H

    2012-09-01

    Powdered maize starch thickeners are used to modify drink consistency in the clinical management of dysphagia. Amylase is a digestive enzyme found in saliva which breaks down starch. This action is dependent on pH, which varies in practice depending on the particular drink. This study measured the effects of human saliva on the viscosity of drinks thickened with a widely used starch-based thickener. Experiments simulated a possible clinical scenario whereby saliva enters a cup and contaminates a drink. Citric acid (E330) was added to water to produce a controlled range of pH from 3.0 to 7.0, and several commercially available drinks with naturally low pH were investigated. When saliva was added to thickened water, viscosity was reduced to less than 1% of its original value after 10-15 min. However, lowering pH systematically slowed the reduction in viscosity attributable to saliva. At pH 3.5 and below, saliva was found to have no significant effect on viscosity. The pH of drinks in this study ranged from 2.6 for Coca Cola to 6.2 for black coffee. Again, low pH slowed the effect of saliva. For many popular drinks, having pH of 3.6 or less, viscosity was not significantly affected by the addition of saliva.

  3. Development of Online Spectroscopic pH Monitoring for Nuclear Fuel Reprocessing Plants: Weak Acid Schemes.

    PubMed

    Casella, Amanda J; Ahlers, Laura R H; Campbell, Emily L; Levitskaia, Tatiana G; Peterson, James M; Smith, Frances N; Bryan, Samuel A

    2015-05-19

    In nuclear fuel reprocessing, separating trivalent minor actinides and lanthanide fission products is extremely challenging and often necessitates tight pH control in TALSPEAK (Trivalent Actinide-Lanthanide Separation by Phosphorus reagent Extraction from Aqueous Komplexes) separations. In TALSPEAK and similar advanced processes, aqueous pH is one of the most important factors governing the partitioning of lanthanides and actinides between an aqueous phase containing a polyaminopolycarboxylate complexing agent and a weak carboxylic acid buffer and an organic phase containing an acidic organophosphorus extractant. Real-time pH monitoring would significantly increase confidence in the separation performance. Our research is focused on developing a general method for online determination of the pH of aqueous solutions through chemometric analysis of Raman spectra. Spectroscopic process-monitoring capabilities, incorporated in a counter-current centrifugal contactor bank, provide a pathway for online, real-time measurement of solution pH. The spectroscopic techniques are process-friendly and can be easily configured for online applications, whereas classic potentiometric pH measurements require frequent calibration/maintenance and have poor long-term stability in aggressive chemical and radiation environments. Raman spectroscopy discriminates between the protonated and deprotonated forms of the carboxylic acid buffer, and the chemometric processing of the Raman spectral data with PLS (partial least-squares) regression provides a means to quantify their respective abundances and therefore determine the solution pH. Interpretive quantitative models have been developed and validated under a range of chemical composition and pH conditions using a lactic acid/lactate buffer system. The developed model was applied to new spectra obtained from online spectral measurements during a solvent extraction experiment using a counter-current centrifugal contactor bank. The model

  4. Acidic pH increases the avidity of FcγR for immune complexes

    PubMed Central

    López, D H; Trevani, A S; Salamone, G; Andonegui, G; Raiden, S; Giordano, M; Geffner, J R

    1999-01-01

    The interaction of immunoglobulin G (IgG) antibodies with FcγR constitutes a critical mechanism through which IgG antibody effector functions are mediated. In the current work we have examined whether human neutrophil FcγR exhibit pH dependence in their association with IgG. Binding assays were performed in culture medium adjusted to different pH values. It was found that the binding of either heat‐aggregated human IgG (AIgG), soluble immune complexes (sIC) or IgG‐coated erythrocytes (IgG‐E) was markedly higher at pH 6·5 than at pH 7·3. This effect was not observed when saturation of FcγR was achieved, suggesting that acidic pH increases the avidity of FcγR for IC without modifying the total binding capacity. Similar results were observed for the binding of AIgG to either monocytes, natural killer (NK) or K562 cells, suggesting that acidic pH increases the avidity of both, FcγRII and FcγRIII. Additional experiments were performed to analyse whether the binding of IgG to FcγRI also showed pH dependence. To this aim, we employed interferon‐γ‐treated human neutrophils and mouse inflammatory macrophages, previously incubated with blocking antibodies directed to FcγRII and FcγRIII. Acidic pH did not enhance the binding of AIgG nor monomeric IgG under these experimental conditions. Further studies are required to determine whether the enhancement of FcγR avidity for IC could be attributed to titration of histidine(s) residues on the Fc fragment of IgG. PMID:10583607

  5. Optimization of pH values to formulate the bireagent kit for serum uric acid assay.

    PubMed

    Huang, Ya; Chen, Yuanxiang; Yang, Xiaolan; Zhao, Hua; Hu, Xiaolei; Pu, Jun; Liao, Juan; Long, Gaobo; Liao, Fei

    2015-01-01

    A new formulation of the bireagent kit for serum uric acid assay was developed based on the effects of pH on enzyme stability. At 4 °C, half-lives of uricases from Bacillus fastidious and Arthrobacter globiforms were longer than 15 months at pH 9.2, but became shorter at pH below 8.0; half-lives of ascorbate oxidase and peroxidase were comparable at pH 6.5 and 7.0, but became much shorter at pH higher than 7.4. In the new formulation of the bireagent kit, Reagent A contained peroxidase, 4-aminoantipyrine, and ascorbate oxidase in 50 mM phosphate buffer at pH 6.5; Reagent B contained B. fastidious or A. globiforms uricase in 50 mM sodium borate buffer at pH 9.2; Reagents A and B were mixed at 4:1 to produce a final pH from 7.2 to 7.6 for developing a stable color. The new bireagent kit consumed smaller quantities of three enzymes for the same shelf life. With the new bireagent kit, there were linear responses of absorbance at 546 nm to uric acid up to 34 mM in reaction mixtures and a good correlation of uric acid levels in clinical sera with those by a commercial kit, but stronger resistance to ascorbate. Therefore, the new formulation was advantageous. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  6. Uric acid plasma level and urine pH in rats treated with ambroxol.

    PubMed

    Drewa, Tomasz; Wolski, Zbigniew; Gruszka, Marzena; Misterek, Bartosz; Lysik, Joanna

    2007-01-01

    It was a chance discovery that ambroxol parenteral administration led to urinary bladder stone formation in rats. This study was undertaken to examine the serum uric acid levels and urine pH in rats after ambroxol parenteral treatment. Ambroxol influence on the uric acid level was measured in 5 rats (Rattus sp.) treated with 60 mg/kg (dissolved in injection water, sc, daily) during 2 weeks. Ambroxol influence on urine pH was examined on 45 rats divided into 3 groups. Rats from the 1st and 2nd group received 30 and 60 mg/kg/24h ambroxol, respectively. Urine was collected once daily and measured with strip kit. All values were presented as the means with standard deviations. The Student t test was used to compare the means, p < 0.05 was considered as significant. Dynamics of pH changes was measured in 4 rats treated with 60 mg/kg/24h of ambroxol. Controls received 1 mL of injection water sc. Serum uric acid level increased up to 8.7 +/- 1.0 mg/dL vs. 5.7 +/- 1.0 mg/dL in control (p < 0.002). In the 1st and 2nd group urine pH increased up to 7.5 +/- 0.5 and 7.6 +/- 0.5 vs. 6.7 +/- 0.4 (p < 0.05). Ambroxol withdrawal resulted in sequential urine pH decrease. 11 days after interruption of ambroxol therapy pH reached the starting value. Urine pH changes and possible disturbances in uric acid metabolic pathway may influence on the stone formation in rats after ambroxol parenteral treatment. The influence of ambroxol on urinary tract GAG layer and the balance between xanthine and CaOx in the urine should be checked.

  7. Vaginal pH and Microbicidal Lactic Acid When Lactobacilli Dominate the Microbiota

    PubMed Central

    O’Hanlon, Deirdre E.; Moench, Thomas R.; Cone, Richard A.

    2013-01-01

    Lactic acid at sufficiently acidic pH is a potent microbicide, and lactic acid produced by vaginal lactobacilli may help protect against reproductive tract infections. However, previous observations likely underestimated healthy vaginal acidity and total lactate concentration since they failed to exclude women without a lactobacillus-dominated vaginal microbiota, and also did not account for the high carbon dioxide, low oxygen environment of the vagina. Fifty-six women with low (0-3) Nugent scores (indicating a lactobacillus-dominated vaginal microbiota) and no symptoms of reproductive tract disease or infection, provided a total of 64 cervicovaginal fluid samples using a collection method that avoided the need for sample dilution and rigorously minimized aerobic exposure. The pH of samples was measured by microelectrode immediately after collection and under a physiological vaginal concentration of CO2. Commercial enzymatic assays of total lactate and total acetate concentrations were validated for use in CVF, and compared to the more usual HPLC method. The average pH of the CVF samples was 3.5 ± 0.3 (mean ± SD), range 2.8-4.2, and the average total lactate was 1.0% ± 0.2% w/v; this is a five-fold higher average hydrogen ion concentration (lower pH) and a fivefold higher total lactate concentration than in the prior literature. The microbicidal form of lactic acid (protonated lactic acid) was therefore eleven-fold more concentrated, and a markedly more potent microbicide, than indicated by prior research. This suggests that when lactobacilli dominate the vaginal microbiota, women have significantly more lactic acid-mediated protection against infections than currently believed. Our results invite further evaluations of the prophylactic and therapeutic actions of vaginal lactic acid, whether provided in situ by endogenous lactobacilli, by probiotic lactobacilli, or by products that reinforce vaginal lactic acid. PMID:24223212

  8. Vaginal pH and microbicidal lactic acid when lactobacilli dominate the microbiota.

    PubMed

    O'Hanlon, Deirdre E; Moench, Thomas R; Cone, Richard A

    2013-01-01

    Lactic acid at sufficiently acidic pH is a potent microbicide, and lactic acid produced by vaginal lactobacilli may help protect against reproductive tract infections. However, previous observations likely underestimated healthy vaginal acidity and total lactate concentration since they failed to exclude women without a lactobacillus-dominated vaginal microbiota, and also did not account for the high carbon dioxide, low oxygen environment of the vagina. Fifty-six women with low (0-3) Nugent scores (indicating a lactobacillus-dominated vaginal microbiota) and no symptoms of reproductive tract disease or infection, provided a total of 64 cervicovaginal fluid samples using a collection method that avoided the need for sample dilution and rigorously minimized aerobic exposure. The pH of samples was measured by microelectrode immediately after collection and under a physiological vaginal concentration of CO2. Commercial enzymatic assays of total lactate and total acetate concentrations were validated for use in CVF, and compared to the more usual HPLC method. The average pH of the CVF samples was 3.5 ± 0.3 (mean ± SD), range 2.8-4.2, and the average total lactate was 1.0% ± 0.2% w/v; this is a five-fold higher average hydrogen ion concentration (lower pH) and a fivefold higher total lactate concentration than in the prior literature. The microbicidal form of lactic acid (protonated lactic acid) was therefore eleven-fold more concentrated, and a markedly more potent microbicide, than indicated by prior research. This suggests that when lactobacilli dominate the vaginal microbiota, women have significantly more lactic acid-mediated protection against infections than currently believed. Our results invite further evaluations of the prophylactic and therapeutic actions of vaginal lactic acid, whether provided in situ by endogenous lactobacilli, by probiotic lactobacilli, or by products that reinforce vaginal lactic acid.

  9. Aspergillus protein degradation pathways with different secreted protease sets at neutral and acidic pH.

    PubMed

    Sriranganadane, Dev; Waridel, Patrice; Salamin, Karine; Reichard, Utz; Grouzmann, Eric; Neuhaus, Jean-Marc; Quadroni, Manfredo; Monod, Michel

    2010-07-02

    Aspergillus fumigatus grows well at neutral and acidic pH in a medium containing protein as the sole nitrogen source by secreting two different sets of proteases. Neutral pH favors the secretion of neutral and alkaline endoproteases, leucine aminopeptidases (Laps) which are nonspecific monoaminopeptidases, and an X-prolyl dipeptidase (DppIV). Acidic pH environment promotes the secretion of an aspartic endoprotease of pepsin family (Pep1) and tripeptidyl-peptidases of the sedolisin family (SedB and SedD). A novel prolyl peptidase, AfuS28, was found to be secreted in both alkaline and acidic conditions. In previous studies, Laps were shown to degrade peptides from their N-terminus until an X-Pro sequence acts as a stop signal. X-Pro sequences can be then removed by DppIV, which allows Laps access to the following residues. We have shown that at acidic pH Seds degrade large peptides from their N-terminus into tripeptides until Pro in P1 or P'1 position acts as a stop for these exopeptidases. However, X-X-Pro and X-X-X-Pro sequences can be removed by AfuS28 thus allowing Seds further sequential proteolysis. In conclusion, both alkaline and acidic sets of proteases contain exoprotease activity capable of cleaving after proline residues that cannot be removed during sequential digestion by nonspecific exopeptidases.

  10. Nestedness in Arbuscular Mycorrhizal Fungal Communities along Soil pH Gradients in Early Primary Succession: Acid-Tolerant Fungi Are pH Generalists.

    PubMed

    Kawahara, Ai; An, Gi-Hong; Miyakawa, Sachie; Sonoda, Jun; Ezawa, Tatsuhiro

    2016-01-01

    Soil acidity is a major constraint on plant productivity. Arbuscular mycorrhizal (AM) fungi support plant colonization in acidic soil, but soil acidity also constrains fungal growth and diversity. Fungi in extreme environments generally evolve towards specialists, suggesting that AM fungi in acidic soil are acidic-soil specialists. In our previous surveys, however, some AM fungi detected in strongly acidic soils could also be detected in a soil with moderate pH, which raised a hypothesis that the fungi in acidic soils are pH generalists. To test the hypothesis, we conducted a pH-manipulation experiment and also analyzed AM fungal distribution along a pH gradient in the field using a synthesized dataset of the previous and recent surveys. Rhizosphere soils of the generalist plant Miscanthus sinensis were collected both from a neutral soil and an acidic soil, and M. sinensis seedlings were grown at three different pH. For the analysis of field communities, rhizosphere soils of M. sinensis were collected from six field sites across Japan, which covered a soil pH range of 3.0-7.4, and subjected to soil trap culture. AM fungal community compositions were determined based on LSU rDNA sequences. In the pH-manipulation experiment the acidification of medium had a significant impact on the compositions of the community from the neutral soil, but the neutralization of the medium had no effect on those of the community from the acidic soil. Furthermore, the communities in lower -pH soils were subsets of (nested in) those in higher-pH soils. In the field communities a significant nestedness pattern was observed along the pH gradient. These observations suggest that the fungi in strongly acidic soils are pH generalists that occur not only in acidic soil but also in wide ranges of soil pH. Nestedness in AM fungal community along pH gradients may have important implications for plant community resilience and early primary succession after disturbance in acidic soils.

  11. Nestedness in Arbuscular Mycorrhizal Fungal Communities along Soil pH Gradients in Early Primary Succession: Acid-Tolerant Fungi Are pH Generalists

    PubMed Central

    Kawahara, Ai; An, Gi-Hong; Miyakawa, Sachie; Sonoda, Jun

    2016-01-01

    Soil acidity is a major constraint on plant productivity. Arbuscular mycorrhizal (AM) fungi support plant colonization in acidic soil, but soil acidity also constrains fungal growth and diversity. Fungi in extreme environments generally evolve towards specialists, suggesting that AM fungi in acidic soil are acidic-soil specialists. In our previous surveys, however, some AM fungi detected in strongly acidic soils could also be detected in a soil with moderate pH, which raised a hypothesis that the fungi in acidic soils are pH generalists. To test the hypothesis, we conducted a pH-manipulation experiment and also analyzed AM fungal distribution along a pH gradient in the field using a synthesized dataset of the previous and recent surveys. Rhizosphere soils of the generalist plant Miscanthus sinensis were collected both from a neutral soil and an acidic soil, and M. sinensis seedlings were grown at three different pH. For the analysis of field communities, rhizosphere soils of M. sinensis were collected from six field sites across Japan, which covered a soil pH range of 3.0–7.4, and subjected to soil trap culture. AM fungal community compositions were determined based on LSU rDNA sequences. In the pH-manipulation experiment the acidification of medium had a significant impact on the compositions of the community from the neutral soil, but the neutralization of the medium had no effect on those of the community from the acidic soil. Furthermore, the communities in lower -pH soils were subsets of (nested in) those in higher-pH soils. In the field communities a significant nestedness pattern was observed along the pH gradient. These observations suggest that the fungi in strongly acidic soils are pH generalists that occur not only in acidic soil but also in wide ranges of soil pH. Nestedness in AM fungal community along pH gradients may have important implications for plant community resilience and early primary succession after disturbance in acidic soils. PMID

  12. Changes in soil pH across England and Wales in response to decreased acid deposition

    NASA Astrophysics Data System (ADS)

    Kirk, G. J. D.; Bellamy, P. H.

    2009-04-01

    In our recent analysis of data from the National Soil Inventory of England and Wales, we found widespread changes in soil pH across both countries between the two samplings of the Inventory. In general, soil pH increased - i.e. soils became less acid - under all land uses. The Inventory was first sampled in 1978-83 on a 5-km grid over the whole area. This yielded about 6,000 sites of which 5,662 could be sampled for soil. Roughly 40% of the sites were re-sampled at intervals from 12 to 25 years after the original sampling - in 1994/96 for agricultural land and in 2002/03 for non-agricultural. Exactly the same sampling and analytical protocols were used in the two samplings. In arable soils, the increase in pH was right across the range, whereas in grassland soils the main increase was at the acid end of the scale (pH < 5.5) with a small increase above pH 7. Some part of the change is likely to have been due to changes in land management. This includes better targeting of agricultural lime on acid soils; changes in nitrogen fertilizer use; deeper ploughing bringing up more calcareous subsoil on soils on calcareous materials; and so forth. However a major driver appears to have been decreased acid deposition to land. The total amounts of nitrogen compounds deposited were relatively unchanged over the survey period, but the amounts of acidifying sulphur compounds decreased by approximately 50%. We constructed a linear regression model to assess the relation between the rate of change in pH (normalised to an annual basis) and the rate of change in acid deposition, as modified by soil properties (pH, clay content, organic matter content), rainfall and past acid deposition. We used data on rainfall and acid deposition over the survey period on the same 5-km grid as the NSI data. We fitted the model separately for each land use category. The results for arable land showed a significant effect of the change in rate of acid deposition, though a significant part of the

  13. The Effects of Volatile Salivary Acids and Bases on Exhaled Breath Condensate pH

    PubMed Central

    Effros, Richard M.; Casaburi, Richard; Su, Jennifer; Dunning, Marshall; Torday, John; Biller, Julie; Shaker, Reza

    2006-01-01

    Rationale: Recent studies have reported acidification of exhaled breath condensate (EBC) in inflammatory lung diseases. This phenomenon, designated “acidopnea,” has been attributed to airway inflammation. Objectives: To determine whether salivary acids and bases can influence EBC pH in chronic obstructive pulmonary disease (COPD). Methods: Measurements were made of pH, electrolytes, and volatile bases and acids in saliva and EBC equilibrated with air in 10 healthy subjects and 10 patients. Results: The average EBC pH in COPD was reduced (normal, 7.24 ± 0.24 SEM; range, 6.11–8.34; COPD, 6.67 ± 0.18; range, 5.74–7.64; p = 0.079). EBCs were well buffered by NH4+/NH3 and CO2/HCO3− in all but four patients, who had NH4+ concentrations under 60 μmol/L, and acetate concentrations that approached or exceeded those of NH4+. Saliva contained high concentrations of acetate (∼ 6,000 μmol/L) and NH4+ (∼ 12,000 μmol/L). EBC acetate increased and EBC NH4+ decreased when salivary pH was low, consistent with a salivary source for these volatile constituents. Nonvolatile acids did not play a significant role in determining pH of condensates because of extreme dilution of respiratory droplets by water vapor (∼ 1:12,000). Transfer of both acetic acid and NH3 from the saliva to the EBC was in the gas phase rather than droplets. Conclusions: EBC acidification in COPD can be affected by the balance of volatile salivary acids and bases, suggesting that EBC pH may not be a reliable marker of airway acidification. Salivary acidification may play an important role in acidopnea. PMID:16284109

  14. A new boronic acid fluorescent sensor based on fluorene for monosaccharides at physiological pH

    NASA Astrophysics Data System (ADS)

    Hosseinzadeh, Rahman; Mohadjerani, Maryam; Pooryousef, Mona; Eslami, Abbas; Emami, Saeed

    2015-06-01

    Fluorescent boronic acids are very useful fluorescent sensor for detection of biologically important saccharides. Herein we synthesized a new fluorene-based fluorescent boronic acid that shows significant fluorescence changes upon addition of saccharides at physiological pH. Upon addition of fructose, sorbitol, glucose, galactose, ribose, and maltose at different concentration to the solution of 7-(dimethylamino)-9,9-dimethyl-9H-fluoren-2-yl-2-boronic acid (7-DMAFBA, 1), significant decreases in fluorescent intensity were observed. It was found that this boronic acid has high affinity (Ka = 3582.88 M-1) and selectivity for fructose over glucose at pH = 7.4. The sensor 1 showed a linear response toward D-fructose in the concentrations ranging from 2.5 × 10-5 to 4 × 10-4 mol L-1 with the detection limit of 1.3 × 10-5 mol L-1.

  15. Polyamine/salt-assembled microspheres coated with hyaluronic acid for targeting and pH sensing.

    PubMed

    Zhang, Pan; Yang, Hui; Wang, Guojun; Tong, Weijun; Gao, Changyou

    2016-06-01

    The poly(allylamine hydrochloride)/trisodium citrate aggregates were fabricated and further covalently crosslinked via the coupling reaction of carboxylic sites on trisodium citrate with the amine groups on polyamine, onto which poly-L-lysine and hyaluronic acid were sequentially assembled, forming stable microspheres. The pH sensitive dye and pH insensitive dye were further labeled to enable the microspheres with pH sensing property. Moreover, these microspheres could be specifically targeted to HeLa tumor cells, since hyaluronic acid can specifically recognize and bind to CD44, a receptor overexpressed on many tumor cells. Quantitative pH measurement by confocal laser scanning microscopy demonstrated that the microspheres were internalized into HeLa cells, and accumulated in acidic compartments. By contrast, only a few microspheres were adhered on the NIH 3T3 cells surface. The microspheres with combined pH sensing property and targeting ability can enhance the insight understanding of the targeted drug vehicles trafficking after cellular internalization.

  16. Dissolved Divalent Metal and pH Effects on Amino Acid Polymerization: A Thermodynamic Evaluation.

    PubMed

    Kitadai, Norio

    2017-03-01

    Polymerization of amino acids is a fundamentally important step for the chemical evolution of life. Nevertheless, its response to changing environmental conditions has not yet been well understood because of the lack of reliable quantitative information. For thermodynamics, detailed prediction over diverse combinations of temperature and pH has been made only for a few amino acid-peptide systems. This study used recently reported thermodynamic dataset for the polymerization of the simplest amino acid "glycine (Gly)" to its short peptides (di-glycine and tri-glycine) to examine chemical and structural characteristics of amino acids and peptides that control the temperature and pH dependence of polymerization. Results showed that the dependency is strongly controlled by the intramolecular distance between the amino and carboxyl groups in an amino acid structure, although the side-chain group role is minor. The polymerization behavior of Gly reported earlier in the literature is therefore expected to be a typical feature for those of α-amino acids. Equilibrium calculations were conducted to examine effects of dissolved metals as a function of pH on the monomer-polymer equilibria of Gly. Results showed that metals shift the equilibria toward the monomer side, particularly at neutral and alkaline pH. Metals that form weak interaction with Gly (e.g., Mg(2+)) have no noticeable influence on the polymerization, although strong interaction engenders significant decrease of the equilibrium concentrations of Gly peptides. Considering chemical and structural characteristics of Gly and Gly peptides that control their interactions with metals, it can be expected that similar responses to the addition of metals are applicable in the polymerization of neutral α-amino acids. Neutral and alkaline aqueous environments with dissolved metals having high affinity with neutral α-amino acids (e.g., Cu(2+)) are therefore not beneficial places for peptide bond formation on the primitive

  17. Dissolved Divalent Metal and pH Effects on Amino Acid Polymerization: A Thermodynamic Evaluation

    NASA Astrophysics Data System (ADS)

    Kitadai, Norio

    2017-03-01

    Polymerization of amino acids is a fundamentally important step for the chemical evolution of life. Nevertheless, its response to changing environmental conditions has not yet been well understood because of the lack of reliable quantitative information. For thermodynamics, detailed prediction over diverse combinations of temperature and pH has been made only for a few amino acid-peptide systems. This study used recently reported thermodynamic dataset for the polymerization of the simplest amino acid "glycine (Gly)" to its short peptides (di-glycine and tri-glycine) to examine chemical and structural characteristics of amino acids and peptides that control the temperature and pH dependence of polymerization. Results showed that the dependency is strongly controlled by the intramolecular distance between the amino and carboxyl groups in an amino acid structure, although the side-chain group role is minor. The polymerization behavior of Gly reported earlier in the literature is therefore expected to be a typical feature for those of α-amino acids. Equilibrium calculations were conducted to examine effects of dissolved metals as a function of pH on the monomer-polymer equilibria of Gly. Results showed that metals shift the equilibria toward the monomer side, particularly at neutral and alkaline pH. Metals that form weak interaction with Gly (e.g., Mg2+) have no noticeable influence on the polymerization, although strong interaction engenders significant decrease of the equilibrium concentrations of Gly peptides. Considering chemical and structural characteristics of Gly and Gly peptides that control their interactions with metals, it can be expected that similar responses to the addition of metals are applicable in the polymerization of neutral α-amino acids. Neutral and alkaline aqueous environments with dissolved metals having high affinity with neutral α-amino acids (e.g., Cu2+) are therefore not beneficial places for peptide bond formation on the primitive

  18. Multistimuli-responsive supramolecular vesicles based on water-soluble pillar[6]arene and SAINT complexation for controllable drug release.

    PubMed

    Cao, Yu; Hu, Xiao-Yu; Li, Yan; Zou, Xiaochun; Xiong, Shuhan; Lin, Chen; Shen, Ying-Zhong; Wang, Leyong

    2014-07-30

    Supramolecular binary vesicles based on the host-guest complexation of water-soluble pillar[6]arene (WP6) and SAINT molecule have been successfully constructed, which showed pH-, Ca(2+)-, and thermal-responsiveness. These supramolecular vesicles can efficiently encapsulate model substrate calcein, which then can be efficiently released either by adjusting the solution pH to acidic condition due to the complete disruption of vesicular structure, or particularly, by adding a certain amount of Ca(2+) due to the Ca(2+)-induced vesicle fusion and accompanied by the structure disruption. More importantly, drug loading and releasing experiments demonstrate that an anticancer drug, DOX, can be successfully encapsulated by the supramolecular vesicles, and the resulting DOX-loaded vesicles exhibit efficient release of the encapsulated DOX with the pH adjustment or the introduction of Ca(2+). Cytotoxicity experiments suggest that the resulting DOX-loaded supramolecular vesicles exhibit comparable therapeutic effect for cancer cells as free DOX and the remarkably reduced damage for normal cells as well. The present multistimuli-responsive supramolecular vesicles have great potential applications in the field of controlled drug delivery. In addition, giant supramolecular vesicles (~3 μm) with large internal volume and good stability can be achieved by increasing the temperature of WP6 ⊃ SAINT vesicular solution, and they might have potential applications for bioimaging.

  19. pH shift protein recovery with organic acids on texture and color of cooked gels.

    PubMed

    Paker, Ilgin; Beamer, Sarah; Jaczynski, Jacek; Matak, Kristen E

    2015-01-01

    Isoelectric solubilization and precipitation (ISP) processing uses pH shifts to separate protein from fish frames, which may increase commercial interest for silver carp. Texture and color properties of gels made from silver carp protein recovered at different pH strategies and organic acid types were compared. ISP was applied to headed gutted silver carp using 10 mol L(-1) sodium hydroxide (NaOH) and either glacial acetic acid (AA) or a (1:1) formic and lactic acid combination (F&L). Protein gels were made with recovered protein and standard functional additives. Texture profile analysis and the Kramer shear test showed that protein gels made from protein solubilized at basic pH values were firmer, harder, more cohesive, gummier and chewier (P < 0.05) than proteins solubilized under acidic conditions. Acidic solubilization led to whiter (P < 0.05) gels, and using F&L during ISP yielded whiter gels under all treatments (P < 0.05). Gels made from ISP-recovered silver carp protein using organic acids show potential for use as a functional ingredient in restructured foods. © 2014 Society of Chemical Industry.

  20. pH responsive poly amino-acid hydrogels formed via silk sericin templating.

    PubMed

    Kurland, Nicholas E; Ragland, Robert B; Zhang, Aolin; Moustafa, Mahmoud E; Kundu, Subhas C; Yadavalli, Vamsi K

    2014-09-01

    Poly(amino acid) hydrogels have attracted a great deal of attention as biodegradable biomaterials that can limit products of synthetic polymer degradation. Here we report on a stimuli-responsive, porous, composite biomaterial based on the protein templating of the poly(amino acid) hydrogel from poly(aspartic acid) with the silk protein sericin. This low-cost, biocompatible and biodegradable hydrogel demonstrates a greatly increased porosity and improvement in volumetric swelling over networks formed from pure poly(aspartic acid). The swelling capacity measured over a range of pH values surrounding physiological pH 7.0 demonstrates a linear profile, in which hydrogel volume and mass increase to a maximum, with an increase as a function of higher sericin content. In comparison to pure poly(aspartic acid), this demonstrates a nearly 3-fold increase in retention volume at basic pH. The increase in swelling is also demonstrated by the increase in porosity and internal micro-architecture of the hydrogel networks. The biomaterial is then shown to perform well as a scaffold for cells with high mechanical strength and integrity. This protein- and homo poly(amino acid)-based super-swelling hydrogel has applications in drug delivery and tissue engineering as an economical and environmentally friendly biomaterial, in addition to ensuring the species incorporated maintain their biocompatibility during processing.

  1. Reconstitution of lactate proton symport activity in plasma membrane vesicles from the yeast Candida utilis.

    PubMed

    Gerós, H; Cássio, F; Leão, C

    1996-09-30

    Lactic acid transport was studied in plasma membrane vesicles from the yeast Candida utilis IGC 3092 which were fused with liposomes containing cytochrome c oxidase. After the addition of an electron donor system, these hybrid membrane vesicles were able to generate a proton-motive force of about--150 mV, inside alkaline and negative. In vesicles prepared from lactic acid-grown cells, the uptake of labelled lactic acid, at pH 6.2, under energized conditions, was expressed by a kinetics consistent with the involvement of a mediated transport system. This carrier exhibited a substrate specificity pattern identical to the one found for the lactate-proton symport in intact cells. The transport of labelled lactic acid was accumulative and strongly sensitive to the effects of the protonophore carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone, consistent with the involvement of the proton-motive force in acid uptake, hence with the presence of a proton symport for lactate. Dissipation of the transmembrane electric potential by valinomycin did not have a significant effect on lactate accumulation, whereas abolishing the transmembrane pH gradient (delta pH) by nigericin prevented the accumulation and led to a rapid efflux of the accumulated acid. The data support that the delta pH is the main component of the proton-motive force involved in the transport of the acid and its accumulation. The lactate-proton symport stoichiometry was 1:1, being independent of the pH. Vesicles prepared from glucose-grown cells did not display the capacity to transport and accumulate lactate. However, activity for the carrier was also reconstituted in vesicles obtained from glucose-grown cells after incubation in buffer containing lactic acid. These results were consistent with those obtained in intact cells, which demonstrated that the lactate-proton symport of the yeast C. utilis is inducible.

  2. Influence of sodium chloride, pH, and lactic acid bacteria on anaerobic lactic acid utilization during fermented cucumber spoilage

    USDA-ARS?s Scientific Manuscript database

    Cucumbers are preserved commercially by natural fermentations in 5% to 8% sodium chloride (NaCl) brines. Occasionally, fermented cucumbers spoil after the primary fermentation is complete. This spoilage has been characterized by decreases in lactic acid and a rise in brine pH caused by microbial ins...

  3. pH, cholesterol sulfate, and fatty acids affect the stratum corneum lipid organization.

    PubMed

    Bouwstra, J A; Gooris, G S; Dubbelaar, F E; Weerheim, A M; Ponec, M

    1998-08-01

    Lipid mixtures prepared from cholesterol (CHOL), isolated ceramides (CER), and free fatty acids can serve as attractive tools to study the role various stratum corneum (SC) lipids or microenvironmental conditions play in the SC lipid organization, as the phase behavior in these mixtures and in SC are similar: two lamellar phases with periodicities of approximately 6 and 13 nm are present. Because pH and cholesterol sulfate (CSO4) gradients exist in SC and may affect the local SC lipid organization, the effects of pH and CSO4 on lipid phase behavior was examined. X-ray diffraction studies with CHOL:CER mixtures revealed that the lamellar ordering at pH 5 and 7.4 were similar: both the short and the long periodicity phases were present. Upon addition of free fatty acids the phase behavior became pH dependent; the long periodicity phase being more dominant at pH 7.4 than at pH 5. Similar observations have been made upon addition of CSO4. Furthermore, only in the presence of CSO4 did phase-separated CHOL disappear, indicating that CHOL completely dissolves in the lamellar phases. A major phase change from an hexagonal to an orthorhombic lateral packing has been observed in the presence of free fatty acids. Furthermore, in the presence of CSO4 next to orthorhombic also liquid lateral packing could be detected. In contrast to lamellar ordering, changes in pH did not affect the lateral packing in any of the lipid mixtures studied.

  4. A colorimetric pH indicators and boronic acids ensemble array for quantitative sugar analysis.

    PubMed

    Ghosh, Krishna Kanta; Yap, Eunice; Kim, Hanjo; Lee, Jun-Seok; Chang, Young-Tae

    2011-04-07

    The colorimetric response patterns of pH indicators and boronic acids ensemble array were used to analyze serial concentrations of mono-, disaccharides quantitatively. Furthermore, this ensemble array was successfully applied to quantify the sugar content in clinically used saline solutions.

  5. Similar bacterial community composition in acidic mining lakes with different pH and lake chemistry.

    PubMed

    Kampe, Heike; Dziallas, Claudia; Grossart, Hans-Peter; Kamjunke, Norbert

    2010-10-01

    As extreme environmental conditions strongly affect bacterial community composition (BCC), we examined whether differences in pH-even at low pH-and in iron and sulfate concentrations lead to changes in BCC of acidic mining lakes. Thereby, we tested the following hypotheses: (1) diversity of the bacterial community in acidic lakes decreases with reducing pH, (2) BCC differs between epilimnion and hypolimnion, and (3) BCC in extremely acidic environments does not vary much over time. Therefore, we investigated the BCC of three acidic lakes with different pH values (2.3, 2.7, and 3.2) by denaturing gradient gel electrophoresis (DGGE) and subsequent sequencing of DGGE bands as well as catalyzed reporter deposition-FISH (CARD-FISH). BCC did not significantly vary among the studied lakes nor differ much between water layers. In contrast, BCC significantly changed over time, which is contradictory to our hypotheses. Bacterial communities were dominated by Alpha-, Beta-, and Gammaproteobacteria, whereas Actino- and Acidobacteria rarely occurred. Cell numbers of both free and attached bacteria were positively related to DOC concentration. Overall, low pH and extreme chemical conditions of the studied lakes led to similar assemblages of bacteria with pronounced temporal differences. This notion indicates that temporal changes in environmental conditions including food web structure also affect unique communities of bacteria thriving at low pH.

  6. Autoinducer-2 detection among commensal oral streptococci is dependent on pH and boric acid.

    PubMed

    Cuadra, Giancarlo A; Frantellizzi, Ashley J; Gaesser, Kimberly M; Tammariello, Steven P; Ahmed, Anika

    2016-07-01

    Autoinducer-2, considered a universal signaling molecule, is produced by many species of bacteria; including oral strains. Structurally, autoinducer-2 can exist bound to boron (borated autoinducer-2). Functionally, autoinducer-2 has been linked to important bacterial processes such as virulence and biofilm formation. In order to test production of autoinducer-2 by a given bacterial strain, a bioassay using marine bioluminescent bacteria Vibrio harveyi as a reporter for autoinducer-2 has been designed. We hypothesize that pH adjustment and addition of boron are required for optimal bioluminescence and accurate autoinducer-2 detection. Using this reporter strain we tested autoinducer-2 activity from two oral commensal species, Streptococcus gordonii DL1 and Streptococcus oralis 34. Spent broth was collected and adjusted to pH 7.5 and supplemented with boric acid prior to measuring autoinducer- 2 activity. Results show that low pH inhibits bioluminescence of the reporter strain, but pH 7.5 allows for bioluminescence induction and proper readings of autoinducer-2 activity. Addition of boric acid also has a positive effect on bioluminescence allowing for a more sensitive detection of autoinducer-2 activity. Our data suggests that although autoinducer-2 is present in spent broth, low pH and/or low levels of boric acid become an obstacle for proper autoinducer-2 detection. For proper autoinducer-2 detection, we propose a protocol using this bioassay to include pH adjustment and boric acid addition to spent broth. Studies on autoinducer-2 activity in several bacteria species represent an important area of study as this universal signaling molecule is involved in critical bacterial phenotypes such as virulence and biofilm formation.

  7. [Effect of pH and fermentation time on yield and optical purity of lactic acid from kitchen wastes fermentation].

    PubMed

    Zhang, Bo; He, Pin-Jing; Shao, Li-Ming

    2007-04-01

    Batch experiments were carried out to analyze the effect of pH and fermentation time on the yield of total lactic acid and the distribution of L- and D-lactic acid among total lactic acid during the non-sterilized fermentation of kitchen wastes. The results show that the concentration of reduced sugar (calculated as organic carbon) is low, and its concentration was higher at neutral and alkali conditions (pH 6 - 8) than at acidic conditions (non-controlled pH and pH = 5). The maximum total lactic acid production rate and yield is 0.59 g x (L x h)(-1) and 0.62 g per gram VS at pH 7, respectively. The proportion of lactic acid (calculated as organic carbon) among the TOC reaches 78% and 89% at controlled pH 7 and 8, respectively. The L-lactic acid is the predominant isomer form at pH 8. Lactic acid concentration depends on pH, fermentation time and interaction from the response surface analysis. pH and fermentation time have a significant effect on the optical purity of lactic acid. At acidic conditions, the ratio of L-lactic acid to the total lactic acid increases with the fermentation time before 120 h, and the ratio reaches 0.9 at 120 h. At alkaline conditions, the ratio keeps at above 0.86 in the whole experimental fermentation time and reachs the maximum value (0.93) at 48 h. It decreases with fermentation time at pH 7. To obtain high lactic acid yield and optical purity simultaneously, it is suggested that pH should be contralled at 8.

  8. Bilayers and wormlike micelles at high pH in fatty acid soap systems.

    PubMed

    Xu, Wenlong; Liu, Huizhong; Song, Aixin; Hao, Jingcheng

    2016-03-01

    Bilayers at high pH in the fatty acid systems of palmitic acid/KOH/H2O, palmitic acid/CsOH/H2O, stearic acid/KOH/H2O and stearic acid/CsOH/H2O can form spontaneously (Xu et al., 2014, 2015). In this work, the bilayers can still be observed at 25°C with an increase of the concentration of fatty acids. We found that wormlike micelles can also be prepared in the fatty acid soap systems at high pH, even though the temperature was increased to be 50°C. The viscoelasticity, apparent viscosity, yield stress of the bilayers were determined by the rheological measurements. Wormlike micelles were identified by cryogenic transmission electron microscopy (cryo-TEM) and emphasized by the rheological characterizations, which are in accordance with the Maxwell fluids with good fit of Cole-Cole plots. The phase transition temperature was determined by differential scanning calorimetry (DSC) and the transition process was recorded. The regulating role of counterions of fatty acids were discussed by (CH3)4N(+), (C2H5)4N(+), (C3H7)4N(+), and (C4H9)4N(+) as comparison, concluding that counterions with appropriate hydrated radius were the vital factor in the formation wormlike micelles. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Evaluation of net acid generation pH as a single indicator for acid forming potential of rocks using geochemical properties.

    PubMed

    Oh, Chamteut; Ji, Sangwoo; Yim, Giljae; Cheong, Youngwook

    2017-04-01

    The main purpose of this research was to evaluate the geochemical properties of rocks for a single indicator of acid-forming potential. The indicators, such as net acid generation (NAG), NAG pH and total S, were applied to 312 rock samples of various geological characteristics. Additional indicators, such as a Modified NAG pH, paste pH and available acid neutralizing capacity (ANC), were applied to 22 selected samples. Among them, NAG pH was considered the most plausible single indicator in evaluating acid-forming potential, as it is simple to measure, widely applicable to various samples and can be used to estimate the NAG value. The acid-forming potential of 287 samples (92% of samples examined in this research) was classified as either non-acid forming (NAF) or potentially acid forming (PAF) by NAG pH, with an NAF criteria of <3.21 and PAF of >4.52. The NAG pH was also a good estimate of the risk of short-term acid release when combined with paste pH information. However, application of NAG pH to coal mine wastes, with high organic carbon contents, produced erroneous results due to the generation of organic acid during the NAG test. In this research, a Modified NAG pH was assessed as an alternative to NAG pH in such situations.

  10. Recovery of carboxylic acids at pH greater than pKa

    SciTech Connect

    Tung, Lisa A.

    1993-08-01

    Economics of producing carboxylic acids by fermentation is often dominated, not by the fermentation cost, but by the cost of recovering and purifying the acids from dilute aqueous solutions. Experiments were performed to measure uptakes of lactic and succinic acids as functions of pH by basic polymeric sorbents; sorbent regeneration was also tested. Performance at pH > pKa and regenerability depend on sorbent basicity; apparent pKa and monomer pK{sub a} can be used to predict sorbent performance. Two basic amine extractants, Alamine 336 and Amberlite LA-2, in were also studied; they are able to sustain capacity to higher pH in diluents that stabilize the acid-amine complex through H bonding. Secondary amines perform better than tert-amines in diluents that solvate the additional proton. Competitive sulfate and phosphate, an interference in fermentation, are taken up by sorbents more strongly than by extractants. The third step in the proposed fermentation process, the cracking of the trimethylammonium (TMA) carboxylate, was also examined. Because lactic acid is more soluble and tends to self-esterify, simple thermal cracking does not remove all TMA; a more promising approach is to esterify the TMA lactate by reaction with an alcohol.

  11. Algal and Bacterial Activities in Acidic (pH 3) Strip Mine Lakes

    PubMed Central

    Gyure, Ruth A.; Konopka, Allan; Brooks, Austin; Doemel, William

    1987-01-01

    Reservoir 29 and Lake B are extremely acid lakes (epilimnion pHs of 2.7 and 3.2, respectively), because they receive acidic discharges from coal refuse piles. They differ in that the pH of profundal sediments in Reservoir 29 increased from 2.7 to 3.8 during the period of thermal stratification, whereas permanently anoxic sediments in Lake B had a pH of 6.2. The pH rise in Reservoir 29 sediments was correlated with a temporal increase in H2S concentration in the anaerobic hypolimnion from 0 to >1 mM. The chlorophyll a levels in the epilimnion of Reservoir 29 were low, and the rate of primary production was typical of an oligotrophic system. However, there was a dense 10-cm layer of algal biomass at the bottom of the metalimnion. Production by this layer was low owing to light limitation and possibly H2S toxicity. The specific photosynthetic rates of epilimnetic algae were low, which suggests that nutrient availability is more important than pH in limiting production. The highest photosynthetic rates were obtained in water samples incubated at pH 2.7 to 4. Heterotrophic bacterial activity (measured by [14C]glucose metabolism) was greatest at the sediment/water interface. Bacterial production (assayed by thymidine incorporation) was as high in Reservoir 29 as in a nonacid mesotrophic Indiana lake. PMID:16347430

  12. Algal and bacterial activities in acidic (pH 3) strip mine lakes

    SciTech Connect

    Gyure, R.A.; Konopka, A.; Brooks, A.; Doemel, W.

    1987-09-01

    Reservoir 29 and Lake B are extremely acid lakes (epilimnion pHs of 2.7 and 3.2, respectively), because they receive acidic discharges from coal refuse piles. They differ in that the pH of profundal sediments in Reservoir 29 increased from 2.7 to 3.8 during the period of thermal stratification, whereas permanently anoxic sediments in Lake B had a pH of 6.2. The pH rise in Reservoir 29 sediments was correlated with a temporal increase in H/sub 2/S concentration in the anaerobic hypolimnion from 0 to >1 mM. The chlorophyll a levels in the epilimnion of Reservoir 29 were low, and the rate of primary production was typical of an oligotrophic system. However, there was a dense 10-cm layer of algal biomass at the bottom of the metalimnion. Production by this layer was low owing to light limitation and possibly H/sub 2/S toxicity. The specific photosynthetic rates of epilimnetic algae were low, which suggests that nutrient availability is more important than pH in limiting production. The highest photosynthetic rates were obtained in water samples incubated at pH 2.7 to 4. Heterotrophic bacterial activity (measured by (/sup 14/C)glucose metabolism) was greatest at the sediment/water interface. Bacterial production (assayed by thymidine incorporation) was as high in Reservoir 29 as in a nonacid mesotrophic Indiana lake.

  13. Effects of amiloride on thermosensitivity of Chinese hamster cells under neutral and acidic pH

    SciTech Connect

    Miyakoshi, J.; Oda, W.; Hirata, M.; Fukuhori, N.; Inagaki, C.

    1986-04-01

    The modifying effects of amiloride on the thermosensitivity of Chinese hamster V-79 cells were examined under both neutral (pH 7.3) and acidic (pH 6.6) conditions. Amiloride, a diuretic drug, is known to inhibit the Na+/H+ exchange activity. Under the extracellular pH of 7.3, amiloride (0.1-0.5 mM) enhanced the thermal cell killing powers of 42/sup 0/C hyperthermia with increasing concentration and exposure time of the drug. The age response of cells to 42/sup 0/C hyperthermia in the presence or absence of amiloride (0.5 mM) showed that amiloride sensitized cells to heat, especially those at G1-S boundary through middle S phases. On the other hand, the lowering of extracellular pH to 6.6 enhanced cell killing by 42/sup 0/C hyperthermia. When cells were exposed to 42/sup 0/C hyperthermia in the presence of amiloride at pH 6.6, cell survival decreased still more. The thermosensitizing effects of the lowered pH at 6.6 and amiloride appeared to be additive. From these results, it is suggested that the thermosensitization by amiloride is probably due, in part, to the inhibition of cellular Na+/H+ exchange activity. The present study proposes the possibility that amiloride may be useful as a hyperthermic sensitizer in a clinical treatment of cancer.

  14. Interaction of Whey Proteins with Phenolic Derivatives Under Neutral and Acidic pH Conditions.

    PubMed

    Cao, Yanyun; Xiong, Youling L

    2017-02-01

    Integration of gallic acid (GA) and its derivative epigallocatechin gallate (EGCG; 20, 120, and 240 μmol/g, protein basis) into whey protein isolate (WPI) at room temperature and pH 3.0 and pH 7.0 was investigated. At pH 7.0, both phenolics caused significant structural changes and EGCG induced greater digestibility of WPI. Total sulfhydryl in WPI decreased from 28.6 to 7.6 μmol/g and surface hydrophobicity declined by nearly 50% with 240 μmol/g EGCG at pH 7.0. Similar but less appreciable changes were produced by GA and at pH 3.0. Isothermal titration and fluorescence quenching tests showed moderately weak binding of WPI with GA but strong binding with EGCG at pH 7.0. Both phenolics at high concentrations lowered the thermal transition temperature of β-lactoglobulin by 0.5 °C to 1.4 °C and promoted its digestion. The phenolics also displayed a remarkable synergism with peptides in the WPI digest exerting radical scavenging activity. © 2017 Institute of Food Technologists®.

  15. Intracellular pH Response to Weak Acid Stress in Individual Vegetative Bacillus subtilis Cells

    PubMed Central

    Pandey, Rachna; Vischer, Norbert O. E.; Smelt, Jan P. P. M.; van Beilen, Johan W. A.; Ter Beek, Alexander; De Vos, Winnok H.; Manders, Erik M. M.

    2016-01-01

    ABSTRACT Intracellular pH (pHi) critically affects bacterial cell physiology. Hence, a variety of food preservation strategies are aimed at perturbing pHi homeostasis. Unfortunately, accurate pHi quantification with existing methods is suboptimal, since measurements are averages across populations of cells, not taking into account interindividual heterogeneity. Yet, physiological heterogeneity in isogenic populations is well known to be responsible for differences in growth and division kinetics of cells in response to external stressors. To assess in this context the behavior of intracellular acidity, we have developed a robust method to quantify pHi at single-cell levels in Bacillus subtilis. Bacilli spoil food, cause disease, and are well known for their ability to form highly stress-resistant spores. Using an improved version of the genetically encoded ratiometric pHluorin (IpHluorin), we have quantified pHi in individual B. subtilis cells, cultured at an external pH of 6.4, in the absence or presence of weak acid stresses. In the presence of 3 mM potassium sorbate, a decrease in pHi and an increase in the generation time of growing cells were observed. Similar effects were observed when cells were stressed with 25 mM potassium acetate. Time-resolved analysis of individual bacteria in growing colonies shows that after a transient pH decrease, long-term pH evolution is highly cell dependent. The heterogeneity at the single-cell level shows the existence of subpopulations that might be more resistant and contribute to population survival. Our approach contributes to an understanding of pHi regulation in individual bacteria and may help scrutinizing effects of existing and novel food preservation strategies. IMPORTANCE This study shows how the physiological response to commonly used weak organic acid food preservatives, such as sorbic and acetic acids, can be measured at the single-cell level. These data are key to coupling often-observed single-cell heterogeneous

  16. [Effects of thiourea on pH and availability of metal ions in acid red soil].

    PubMed

    Yang, Bo; Wang, Wen; Zeng, Qing-Ru; Zhou, Xi-Hong

    2014-03-01

    Through the simulation research, the effects of application of thiourea and urea on pH and availability of metal ions in acid red soil were studied, and the results showed that after applying urea, the soil pH increased in the first experimental stage and then reduced gradually to a low level, however, decreased trends of soil pH values were inhibited by the application of thiourea, especially when the concentration of thiourea reached to 5.0 mmol x kg(-1) dry soil, the soil pH was stable at high level, which exceeded to 6.0. It proved that the application of thiourea could inhibit the soil acidification due to urea application. After applying urea with different concentrations of thiourea, the available contents of Zn and Al decreased with the increasing concentration of thiourea, nevertheless, when the concentration of thiourea reached to 5.0 mmol x kg(-1), the available content of Mn was stable at high level which was over 110 mg x kg(-1). In addition, the results showed a highly significant negative correlation between the soil pH and the available content of Cu, Zn and Al, but for Mn, no discipline was found between the soil pH and the availability after applying thiourea. Moreover, the soil pH became higher after applying urea with thiourea compared to add urea only, which led to the decreasing of available content of Al, and it was benefited for the control of the phytotoxic effect of Al. The available content of Mn in the soil not only depended on soil pH but also the content of thiourea due to its redox and complexing reaction with Mn.

  17. Anoxic Biodegradation of Isosaccharinic Acids at Alkaline pH by Natural Microbial Communities

    PubMed Central

    Rout, Simon P.; Charles, Christopher J.; Doulgeris, Charalampos; McCarthy, Alan J.; Rooks, Dave J.; Loughnane, J. Paul; Laws, Andrew P.; Humphreys, Paul N.

    2015-01-01

    One design concept for the long-term management of the UK’s intermediate level radioactive wastes (ILW) is disposal to a cementitious geological disposal facility (GDF). Under the alkaline (10.0<pH>13.0) anoxic conditions expected within a GDF, cellulosic wastes will undergo chemical hydrolysis. The resulting cellulose degradation products (CDP) are dominated by α- and β-isosaccharinic acids (ISA), which present an organic carbon source that may enable subsequent microbial colonisation of a GDF. Microcosms established from neutral, near-surface sediments demonstrated complete ISA degradation under methanogenic conditions up to pH 10.0. Degradation decreased as pH increased, with β-ISA fermentation more heavily influenced than α-ISA. This reduction in degradation rate was accompanied by a shift in microbial population away from organisms related to Clostridium sporosphaeroides to a more diverse Clostridial community. The increase in pH to 10.0 saw an increase in detection of Alcaligenes aquatilis and a dominance of hydrogenotrophic methanogens within the Archaeal population. Methane was generated up to pH 10.0 with acetate accumulation at higher pH values reflecting a reduced detection of acetoclastic methanogens. An increase in pH to 11.0 resulted in the accumulation of ISA, the absence of methanogenesis and the loss of biomass from the system. This study is the first to demonstrate methanogenesis from ISA by near surface microbial communities not previously exposed to these compounds up to and including pH 10.0. PMID:26367005

  18. Is Your Drinking Water Acidic? A Comparison of the Varied pH of Popular Bottled Waters.

    PubMed

    Wright, Kellie F

    2015-06-01

    Dental professionals continually educate patients on the dangers of consuming acidic foods and beverages due to their potential to contribute to dental erosion and tooth decay. Excess acid in the diet can also lead to acidosis, which causes negative systemic side effects. However, water is not typically categorized as acidic. The purpose of this in-vitro study was to investigate the pH levels of several popular brands of bottled water and compare them to various other acidic beverages. Two different brands of marketed alkaline water (with a pH of 8.8 or higher) were also studied, tested for acidity and described. A pilot in-vitro study was conducted to determine the pH levels of a convenience sample of popular brands of bottled water, tap water and other known acidic beverages in comparison with the pH values reported on the respective manufacturers' website. Each beverage was tested in a laboratory using a calibrated Corning pH meter model 240, and waters were compared to the corresponding company's testified pH value. Waters were also compared and contrasted based on their process of purification. The data was then compiled and analyzed descriptively. The pH values for the tested beverages and bottled waters were found to be predominantly acidic. Ten out of the 14 beverages tested were acidic (pH<7), 2 municipal (or "tap") waters were neutral (pH=7) and 2 bottled waters were alkaline (pH>7). The majority of waters tested had a more acidic pH when tested in the lab than the value listed in their water quality reports. It is beneficial for the health care provider to be aware of the potential acidity of popular bottled drinking waters and educate patients accordingly. Copyright © 2015 The American Dental Hygienists’ Association.

  19. Microenvironmental pH measurement during sodium naproxenate dissolution in acidic medium by UV/vis imaging.

    PubMed

    Ostergaard, Jesper; Jensen, Henrik; Larsen, Susan W; Larsen, Claus; Lenke, Jim

    2014-11-01

    Variable dissolution from sodium salts of drugs containing a carboxylic acid group after passing the acidic environment of the stomach may affect oral bioavailability. The aim of the present proof of concept study was to investigate pH effects in relation to the dissolution of sodium naproxenate in 0.01M hydrochloric acid. For this purpose a UV/vis imaging-based approach capable of measuring microenvironmental pH in the vicinity of the solid drug compact as well as monitoring drug dissolution was developed. Using a pH indicating dye real-time spatially resolved measurement of pH was achieved. Sodium naproxenate, can significantly alter the local pH of the dissolution medium, is eventually neutralized and precipitates as the acidic species naproxen. The developed approach is considered useful for detailed studies of pH dependent dissolution phenomena in dissolution testing. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Development of melamine modified urea formaldehyde resins based o nstrong acidic pH catalyzed urea formaldehyde polymer

    Treesearch

    Chung-Yun Hse

    2009-01-01

    To upgrade the performance of urea-formaldehyde (UF) resin bonded particleboards, melamine modified urea-formaldehyde (MUF) resins based on strong acidic pH catalyzed UF polymers were investigated. The study was conducted in a series of two experiments: 1) formulation of MUF resins based on a UF polymer catalyzed with strong acidic pH and 2) determination of the...

  1. Formation of simple single-tailed vesicles mediated by lipophilic solid surfaces.

    PubMed

    Du, Na; Zhu, Xiaoyu; Song, Ruiying; Song, Shue; Hou, Wanguo

    2016-10-19

    Adsorption and aggregation of surfactants at solid-liquid interfaces were fairly well understood, but there was limited knowledge regarding the effect of the presence of a solid surface on aggregate structures in bulk solution. Except for the fatty acid system, most simple single-tailed surfactants (STSs) are well known to form micelles but not vesicles in aqueous solution. Herein, we report a novel phenomenon: with the mediation of lipophilic solid surfaces (LSSs), the zwitterionic STS lauryl sulfobetaine (LSB) formed vesicles from its micellar solution without any additives, producing a mixed solution of vesicles and micelles. More interestingly, the STS vesicles coexisted stably with micelles in the solution and were thermally insensitive even after the removal of LSSs. The quantity of LSB vesicles decreases with the addition of ethanol. The pH effects (4.0-9.0) did not have an obvious influence on the formation and stability of the LSB vesicles. Similar results were obtained from the other STSs, suggesting that the LSS-mediated micelle-to-vesicle transition may be a general phenomenon. We proposed a possible mechanism that adsorption, the matrix effect, and interdigitated bilayer structures were probably crucial for the formation and stability of STS vesicles. We expect this work to provide important insights into the effect of the solid/liquid interface on the self-assembly chemistry of surfactants in bulk solution.

  2. Monomeric banana lectin at acidic pH overrules conformational stability of its native dimeric form.

    PubMed

    Khan, Javed M; Qadeer, Atiyatul; Ahmad, Ejaz; Ashraf, Raghib; Bhushan, Bharat; Chaturvedi, Sumit K; Rabbani, Gulam; Khan, Rizwan H

    2013-01-01

    Banana lectin (BL) is a homodimeric protein categorized among jacalin-related family of lectins. The effect of acidic pH was examined on conformational stability of BL by using circular dichroism, intrinsic fluorescence, 1-anilino-8-napthalene sulfonate (ANS) binding, size exclusion chromatography (SEC) and dynamic light scattering (DLS). During acid denaturation of BL, the monomerization of native dimeric protein was found at pH 2.0. The elution profile from SEC showed two different peaks (59.65 ml & 87.98 ml) at pH 2.0 while single peak (61.45 ml) at pH 7.4. The hydrodynamic radii (R h) of native BL was 2.9 nm while at pH 2.0 two species were found with R h of 1.7 and 3.7 nm. Furthermore at, pH 2.0 the secondary structures of BL remained unaltered while tertiary structure was significantly disrupted with the exposure of hydrophobic clusters confirming the existence of molten globule like state. The unfolding of BL with different subunit status was further evaluated by urea and temperature mediated denaturation to check their stability. As inferred from high Cm and ΔG values, the monomeric form of BL offers more resistance towards chemical denaturation than the native dimeric form. Besides, dimeric BL exhibited a Tm of 77°C while no loss in secondary structures was observed in monomers even up to 95°C. To the best of our knowledge, this is the first report on monomeric subunit of lectins showing more stability against denaturants than its native dimeric state.

  3. Stabilization of T4 bacteriophage at acidic and basic pH by adsorption on paper.

    PubMed

    Meyer, Abigail; Greene, Melissa; Kimmelshue, Chad; Cademartiri, Rebecca

    2017-09-05

    Bacteriophages find applications in agriculture, medicine, and food safety. Many of these applications can expose bacteriophages to stresses that inactivate them including acidic and basic pH. Bacteriophages can be stabilized against these stresses by materials including paper, a common material in packaging and consumer products. Combining paper and bacteriophages creates antibacterial materials, which can reduce the use of antibiotics. Here we show that adsorption on paper protects T4, T5, and T7 bacteriophage from acidic and basic pH. We added bacteriophages to filter paper functionalized with carboxylic acid (carboxyl methyl cellulose) or amine (chitosan) groups, and exposed them to pH from 5.6 to 14. We determined the number of infective bacteriophages after exposure directly on the paper. All papers extended the lifetime of infective bacteriophage by at least a factor of four with some papers stabilizing bacteriophages for up to one week. The degree of stabilization depended on five main factors (i) the family of the bacteriophage, (ii) the charge of the paper and bacteriophages, (iii) the location of the bacteriophages within the paper, (iv) the ability of the paper to prevent bacteriophage-bacteriophage aggregation, and (v) the sensitivity of the bacteriophage proteins to the tested pH. Even when adsorbed on paper the bacteriophages were able to remove E. coli in milk. Choosing the right paper modification or material will protect bacteriophages adsorbed on that material against detrimental pH and other environmental challenges increasing the range of applications of bacteriophages on materials. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Acidic pH increases airway surface liquid viscosity in cystic fibrosis

    PubMed Central

    Tang, Xiao Xiao; Ostedgaard, Lynda S.; Hoegger, Mark J.; Moninger, Thomas O.; Karp, Philip H.; McMenimen, James D.; Choudhury, Biswa; Varki, Ajit; Stoltz, David A.; Welsh, Michael J.

    2016-01-01

    Cystic fibrosis (CF) disrupts respiratory host defenses, allowing bacterial infection, inflammation, and mucus accumulation to progressively destroy the lungs. Our previous studies revealed that mucus with abnormal behavior impaired mucociliary transport in newborn CF piglets prior to the onset of secondary manifestations. To further investigate mucus abnormalities, here we studied airway surface liquid (ASL) collected from newborn piglets and ASL on cultured airway epithelia. Fluorescence recovery after photobleaching revealed that the viscosity of CF ASL was increased relative to that of non-CF ASL. CF ASL had a reduced pH, which was necessary and sufficient for genotype-dependent viscosity differences. The increased viscosity of CF ASL was not explained by pH-independent changes in HCO3– concentration, altered glycosylation, additional pH-induced disulfide bond formation, increased percentage of nonvolatile material, or increased sulfation. Treating acidic ASL with hypertonic saline or heparin largely reversed the increased viscosity, suggesting that acidic pH influences mucin electrostatic interactions. These findings link loss of cystic fibrosis transmembrane conductance regulator–dependent alkalinization to abnormal CF ASL. In addition, we found that increasing Ca2+ concentrations elevated ASL viscosity, in part, independently of pH. The results suggest that increasing pH, reducing Ca2+ concentration, and/or altering electrostatic interactions in ASL might benefit early CF. PMID:26808501

  5. Acidic pH increases airway surface liquid viscosity in cystic fibrosis.

    PubMed

    Tang, Xiao Xiao; Ostedgaard, Lynda S; Hoegger, Mark J; Moninger, Thomas O; Karp, Philip H; McMenimen, James D; Choudhury, Biswa; Varki, Ajit; Stoltz, David A; Welsh, Michael J

    2016-03-01

    Cystic fibrosis (CF) disrupts respiratory host defenses, allowing bacterial infection, inflammation, and mucus accumulation to progressively destroy the lungs. Our previous studies revealed that mucus with abnormal behavior impaired mucociliary transport in newborn CF piglets prior to the onset of secondary manifestations. To further investigate mucus abnormalities, here we studied airway surface liquid (ASL) collected from newborn piglets and ASL on cultured airway epithelia. Fluorescence recovery after photobleaching revealed that the viscosity of CF ASL was increased relative to that of non-CF ASL. CF ASL had a reduced pH, which was necessary and sufficient for genotype-dependent viscosity differences. The increased viscosity of CF ASL was not explained by pH-independent changes in HCO3- concentration, altered glycosylation, additional pH-induced disulfide bond formation, increased percentage of nonvolatile material, or increased sulfation. Treating acidic ASL with hypertonic saline or heparin largely reversed the increased viscosity, suggesting that acidic pH influences mucin electrostatic interactions. These findings link loss of cystic fibrosis transmembrane conductance regulator-dependent alkalinization to abnormal CF ASL. In addition, we found that increasing Ca2+ concentrations elevated ASL viscosity, in part, independently of pH. The results suggest that increasing pH, reducing Ca2+ concentration, and/or altering electrostatic interactions in ASL might benefit early CF.

  6. A partly folded state of acidic fibroblast growth factor at low pH.

    PubMed

    Sanz, J M; Giménez-Gallego, G

    1997-06-01

    Acid denaturation of acidic fibroblast growth factor (aFGF) at low ionic strength was monitored by far-ultraviolet circular dichroism and intrinsic fluorescence. The two spectroscopic probes displayed non-coincident transitions, which suggested the accumulation of partly folded species around pH 4.0. Although under these conditions the fluorescence of aFGF resembled that of the unfolded form of the protein, far-ultraviolet circular dichroism and proton nuclear magnetic resonance spectra indicated the presence of persistent secondary and tertiary structure. Moreover, at pH 4.0, aFGF showed cooperative thermal denaturation and interacted weakly with the hydrophobic probe N-phenyl-1-naphthylamine, showing a relatively high level of structure that did not fit into the classical molten globule category. This intermediate is also capable of interacting with liposomes and might represent a membrane translocation-competent form.

  7. Effect of pH and organic acids on nitrogen transformations and metal dissolution in soils

    SciTech Connect

    Fu, Minhong.

    1989-01-01

    The effect of pH (4, 6, and 8) on nitrogen mineralization was evaluated in three Iowa surface soils treated with crop residues (corn (Zea mays L.), soybean (Glycine max (L.) Merr.), and sorghum (Sorghum vulgare Pers.), or alfalfa (Medicago sativa L.)) and incubated in leaching columns under aerobic conditions at 30C for 20 weeks. In general, N mineralization was significantly depressed at soil pH 4, compared with pH 6 or 8. The types of crop residues added influenced the pattern and amount of N mineralization. A study on the effect of 19 trace elements on the nitrate red activity of four Iowa surface soils showed that most trace elements inhibited this enzyme in acid and neutral soils. The trace elements Ag(I), Cd(II), Se(IV), As(V), and W(VI) were the most effective inhibitors, with >75% inhibition. Mn(II) was the least effective inhibitor, with <10% inhibition. Other trace elements included Cu(I), Co(II), Cu(II), Fe(II), Ni(II), Pb(II), Zn(II), Al(III), As(III), Cr(III), Fe(III), V(IV), Mo(VI), and Se(VI). The application of high-performance liquid chromatography (HPLC) showed that, when coupled to a refractive index detector, it is a rapid, sensitive, and accurate method for determining organic acids in soils. Three organic acids, acetic (2-20 mM), propionic (0-3 mM), and n-butyric (0-1.4 mM), were identified with HPLC and confirmed by gas chromatography in crop-residue-treated soils incubated under waterlogged conditions at 25C for 72 h. No organic acids were detected under aerobic conditions. Four mineral acids and 29 organic acids were studied for their effect on N mineralization and metal dissolution in soils incubated under waterlogged conditions at 30C for 10 days.

  8. The Synthesis of [gamma]-Aminobutyric Acid in Response to Treatments Reducing Cytosolic pH.

    PubMed

    Crawford, L. A.; Bown, A. W.; Breitkreuz, K. E.; Guinel, F. C.

    1994-03-01

    [gamma]-Aminobutyric acid (GABA) synthesis (L-glutamic acid + H+ -> GABA + CO2) is rapidly stimulated by a variety of stress conditions including hypoxia. Recent literature suggests that GABA production and concomitant H+ consumption ameliorates the cytosolic acidification associated with hypoxia or other stresses. This proposal was investigated using isolated asparagus (Asparagus sprengeri Regel) mesophyll cells. Cell acidification was promoted using hypoxia, H+/L-glutamic acid symport, and addition of butyrate or other permeant weak acids. Sixty minutes of all three treatments stimulated the levels of both intracellular and extracellular GABA by values ranging from 100 to 1800%. At an external pH of 5.0, addition of 5 mM butyrate stimulated an increase in overall GABA level from 3.86 (0.56 [plus or minus] SE) to 20.4 (2.16 [plus or minus] SE) nmol of GABA/106 cell. Butyrate stimulated GABA levels by 200 to 300% within 15 s, and extracellular GABA was observed after 10 min. The acid load due to butyrate addition was assayed by measuring [14C]butyrate uptake. After 45 s of butyrate treatment, H+-consuming GABA production accounted for 45% of the imposed acid load. The cytosolic location of a fluorescent pH probe was confirmed using fluorescent microscopy. Spectrofluorimetry indicated that butyrate addition reduced cytosolic pH by 0.60 units with a half-time of approximately 2 s. The proposal that GABA synthesis ameliorates cytosolic acidification is supported by the data. The possible roles of H+ and Ca2+ in stimulating GABA synthesis are discussed.

  9. Indomethacin inhibits tetrodotoxin-resistant Na(+) channels at acidic pH in rat nociceptive neurons.

    PubMed

    Nakamura, Michiko; Jang, Il-Sung

    2016-06-01

    Non-steroidal anti-inflammatory drugs (NSAIDs) are well-known inhibitors of cyclooxygenases (COXs) and are widely used for the treatment of inflammatory pain; however several NSAIDs display COX-independent analgesic action including the inhibition of voltage-gated Na(+) channels expressed in primary afferent neurons. In the present study, we examined whether NSAIDs modulate tetrodotoxin-resistant (TTX-R) Na(+) channels and if this modulation depends on the extracellular pH. The TTX-R Na(+) currents were recorded from small-sized trigeminal ganglion neurons by using a whole-cell patch clamp technique. Among eight NSAIDs tested in this study, several drugs, including aspirin and ibuprofen, did not affect TTX-R Na(+) channels either at pH 7.4 or at pH 6.0. However, we found that indomethacin, and, to a lesser extent, ibuprofen and naproxen potently inhibited the peak amplitude of TTX-R Na(+) currents at pH 6.0. The indomethacin-induced inhibition of TTX-R Na(+) channels was more potent at depolarized membrane potentials. Indomethacin significantly shifted both the voltage-activation and voltage-inactivation relationships to depolarizing potentials at pH 6.0. Indomethacin accelerated the development of inactivation and retarded the recovery from inactivation of TTX-R Na(+) channels at pH 6.0. Given that indomethacin and several other NSAIDs could further suppress local nociceptive signals by inhibiting TTX-R Na(+) channels at an acidic pH in addition to the classical COX inhibition, these drugs could be particularly useful for the treatment of inflammatory pain. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Chlorogenic acid increased 5-hydroxymethylfurfural formation when heating fructose alone or with aspartic acid at two pH levels.

    PubMed

    Zhang, Zhenhua; Zou, Yueyu; Wu, Taigang; Huang, Caihuan; Pei, Kehan; Zhang, Guangwen; Lin, Xiaohua; Bai, Weibin; Ou, Shiyi

    2016-01-01

    Chlorogenic acid (CGA) is a phenolic acid that ubiquitously exists in fruits. This work aims to investigate whether and how CGA influences HMF formation during heating fructose alone, or with an amino acid. The results showed that that CGA increased 5-hydroxymethylfurfural (HMF) formation. At pH 5.5 and 7.0, the addition of 5.0 μmol/ml CGA increased HMF formation by 49.4% and 25.2%, respectively when heating fructose alone, and by 9.0% and 16.7%, respectively when heating fructose with aspartic acid. CGA significantly increased HMF formation by promoting 3-deoxosone formation, and its conversion to HMF by inhibiting HMF elimination, especially in the Maillard reaction system. A comparison of the catalytic capacity of CGA with its six analogous compounds showed that both its di-hydroxyphenyl and carboxyl groups function in increasing HMF formation.

  11. Role of Organic Anion and Amino Acid Carriers in Transport of Inorganic Mercury in Rat Renal Basolateral Membrane Vesicles: Influence of Compensatory Renal Growth

    PubMed Central

    Lash, Lawrence H.; Hueni, Sarah E.; Putt, David A.; Zalups, Rudolfs K.

    2006-01-01

    Susceptibility to renal injury induced by inorganic mercury (Hg2+) increases significantly as a result of compensatory renal growth (following reductions of renal mass). We hypothesize that this phenomenon is related in part to increased basolateral uptake of Hg2+ by proximal tubular cells. To determine the mechanistic roles of various transporters, we studied uptake of Hg2+, in the form of biologically relevant Hg2+-thiol conjugates, using baso-lateral membrane (BLM) vesicles isolated from the kidney(s) of control and uninephrectomized (NPX) rats. Binding of Hg2+ to membranes, accounted for 52–86% of total Hg2+ associated with membrane vesicles exposed to HgCl2, decreased with increasing concentrations of HgCl2, and decreased slightly in the presence of sodium ions. Conjugation of Hg2+ with thiols (glutathione, l-cysteine (Cys), N-acetyl-l-cysteine) reduced binding by more than 50%. Under all conditions, BLM vesicles from NPX rats exhibited a markedly lower proportion of binding. Of the Hg2+-thiol conjugates studied, transport of Hg-(Cys)2 was fastest. Selective inhibition of BLM carriers implicated the involvement of organic anion transporter(s) (Oat1 and/or Oat3; Slc22a6 and Slc22a8), amino acid transporter system ASC (Slc7a10), the dibasic amino acid transporter (Slc3a1), and the sodium-dicarboxylate carrier (SDCT2 or NADC3; Slc13a3). Uptake of each mercuric conjugate, when factored by membrane protein content, was higher in BLM vesicles from uninephrectomized (NPX) rats, with specific increases in transport by the carriers noted above. These results support the hypothesis that compensatory renal growth is associated with increased uptake of Hg2+ in proximal tubular cells and we have identified specific transporters involved in the process. PMID:16162843

  12. Effects of Oxygen Availability on Acetic Acid Tolerance and Intracellular pH in Dekkera bruxellensis

    PubMed Central

    Capusoni, Claudia; Arioli, Stefania; Zambelli, Paolo; Moktaduzzaman, M.; Mora, Diego

    2016-01-01

    ABSTRACT The yeast Dekkera bruxellensis, associated with wine and beer production, has recently received attention, because its high ethanol and acid tolerance enables it to compete with Saccharomyces cerevisiae in distilleries that produce fuel ethanol. We investigated how different cultivation conditions affect the acetic acid tolerance of D. bruxellensis. We analyzed the ability of two strains (CBS 98 and CBS 4482) exhibiting different degrees of tolerance to grow in the presence of acetic acid under aerobic and oxygen-limited conditions. We found that the concomitant presence of acetic acid and oxygen had a negative effect on D. bruxellensis growth. In contrast, incubation under oxygen-limited conditions resulted in reproducible growth kinetics that exhibited a shorter adaptive phase and higher growth rates than those with cultivation under aerobic conditions. This positive effect was more pronounced in CBS 98, the more-sensitive strain. Cultivation of CBS 98 cells under oxygen-limited conditions improved their ability to restore their intracellular pH upon acetic acid exposure and to reduce the oxidative damage to intracellular macromolecules caused by the presence of acetic acid. This study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can protect against the damage caused by the presence of acetic acid. This aspect is important for optimizing industrial processes performed in the presence of acetic acid. IMPORTANCE This study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can have a protective role against the damage caused by the presence of acetic acid. This aspect is important for the optimization of industrial processes performed in the presence of acetic acid. PMID:27235432

  13. Effects of Oxygen Availability on Acetic Acid Tolerance and Intracellular pH in Dekkera bruxellensis.

    PubMed

    Capusoni, Claudia; Arioli, Stefania; Zambelli, Paolo; Moktaduzzaman, M; Mora, Diego; Compagno, Concetta

    2016-08-01

    The yeast Dekkera bruxellensis, associated with wine and beer production, has recently received attention, because its high ethanol and acid tolerance enables it to compete with Saccharomyces cerevisiae in distilleries that produce fuel ethanol. We investigated how different cultivation conditions affect the acetic acid tolerance of D. bruxellensis We analyzed the ability of two strains (CBS 98 and CBS 4482) exhibiting different degrees of tolerance to grow in the presence of acetic acid under aerobic and oxygen-limited conditions. We found that the concomitant presence of acetic acid and oxygen had a negative effect on D. bruxellensis growth. In contrast, incubation under oxygen-limited conditions resulted in reproducible growth kinetics that exhibited a shorter adaptive phase and higher growth rates than those with cultivation under aerobic conditions. This positive effect was more pronounced in CBS 98, the more-sensitive strain. Cultivation of CBS 98 cells under oxygen-limited conditions improved their ability to restore their intracellular pH upon acetic acid exposure and to reduce the oxidative damage to intracellular macromolecules caused by the presence of acetic acid. This study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can protect against the damage caused by the presence of acetic acid. This aspect is important for optimizing industrial processes performed in the presence of acetic acid. This study reveals an important role of oxidative stress in acetic acid tolerance in D. bruxellensis, indicating that reduced oxygen availability can have a protective role against the damage caused by the presence of acetic acid. This aspect is important for the optimization of industrial processes performed in the presence of acetic acid. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  14. Antimony leaching release from brake pads: Effect of pH, temperature and organic acids.

    PubMed

    Hu, Xingyun; He, Mengchang; Li, Sisi

    2015-03-01

    Metals from automotive brake pads pollute water, soils and the ambient air. The environmental effect on water of antimony (Sb) contained in brake pads has been largely untested. The content of Sb in one abandoned brake pad reached up to 1.62×10(4) mg/kg. Effects of initial pH, temperature and four organic acids (acetic acid, oxalic acid, citric acid and humic acid) on Sb release from brake pads were studied using batch reactors. Approximately 30% (97 mg/L) of the total Sb contained in the brake pads was released in alkaline aqueous solution and at higher temperature after 30 days of leaching. The organic acids tested restrained Sb release, especially acetic acid and oxalic acid. The pH-dependent concentration change of Sb in aqueous solution was best fitted by a logarithmic function. In addition, Sb contained in topsoil from land where brake pads were discarded (average 9×10(3) mg/kg) was 3000 times that in uncontaminated soils (2.7±1 mg/kg) in the same areas. Because potentially high amounts of Sb may be released from brake pads, it is important that producers and environmental authorities take precautions. Copyright © 2015. Published by Elsevier B.V.

  15. Relationships between the resistance of yeasts to acetic, propanoic and benzoic acids and to methyl paraben and pH.

    PubMed

    Warth, A D

    1989-07-01

    Minimum inhibitory concentrations of acetic, propanoic and benzoic acids and methyl paraben were determined at pH 3.50 for 22 isolates of 11 yeast species, differing in their resistance to preservatives. Growth in the presence of benzoic acid enhanced the resistance of yeasts to benzoic and the other weak acid preservatives, but not to methyl paraben. Resistance to acetic, propanoic and benzoic acids was strongly correlated, but was not closely related to resistance to methyl paraben. Minimum pH for growth was not related to resistance to the weak acids. The results suggest that growth in the presence of weak-acid preservatives involves a common resistance mechanism.

  16. Kinetics of hyaluronan hydrolysis in acidic solution at various pH values.

    PubMed

    Tømmeraas, Kristoffer; Melander, Claes

    2008-06-01

    Hyaluronic acid (HA) was hydrolyzed using varying temperatures (40, 60, and 80 degrees C) and acid concentrations (0.0010, 0.010, 0.10, 0.50, 1.0, and 2.0 M HCl). The degradation process was monitored by determination of weight average molecular weight ( M w) by size-exclusion chromatography with online multiangle laser light scattering, refractive index, and intrinsic viscosity detectors (SEC-MALLS-RI-visc) on samples taken out continuously during the hydrolysis. SEC-MALLS-RI-visc showed that the degradation gave narrow molecular weight distributions with polydispersity indexes ( M w/ M n) of 1.3-1.7. Kinetic plots of 1/ M w versus time gave linear plots showing that acid hydrolysis of HA is a random process and that it follows a first order kinetics. For hydrolysis in HCl at 60 and 80 degrees C, it was shown that the kinetic rate constant ( k h) for the degradation depended linearly on the acid concentration. Further, the dependence of temperature on the hydrolysis in 0.1 M HCl was found to give a linear Arrhenius plot (ln k h vs 1/ T), with an activation energy ( E a) of 137 kJ/mol and Arrhenius constant ( A) of 7.86 x 10 (15) h (-1). (1)H NMR spectroscopy was used to characterize the product of extensive hydrolysis (48 h at 60 degrees C in 0.1 M HCl). No indication of de- N-acetylation of the N-acetyl glucosamine (GlcNAc) units or other byproducts were seen. Additionally, a low molecular weight HA was hydrolyzed in 0.1 M DCl for 4 h at 80 degrees C. It was shown that it was primarily the beta-(1-->4)-linkage between GlcNAc and glucuronic acid (GlcA) that was cleaved during hydrolysis at pH < p K a,GlcA. The dependence of the hydrolysis rate constant was further studied as a function of pH between -0.3 and 5. The degradation was found to be random (linear kinetic plots) over the entire pH range studied. Further, the kinetic rate constant was found to depend linearly on pH in the region -0.3 to 3. Above this pH (around the p K a of HA), the kinetic constant

  17. Investigation of pH Influence on Skin Permeation Behavior of Weak Acids Using Nonsteroidal Anti-Inflammatory Drugs.

    PubMed

    Chantasart, Doungdaw; Chootanasoontorn, Siriwan; Suksiriworapong, Jiraphong; Li, S Kevin

    2015-10-01

    As a continuing effort to understand the skin permeation behavior of weak acids and bases, the objectives of the present study were to evaluate skin permeation of nonsteroidal anti-inflammatory drugs (NSAIDs) under the influence of pH, investigate the mechanism of pH effect, and examine a previous hypothesis that the effective skin pH for drug permeation is different from donor solution pH. In vitro permeability experiments were performed in side-by-side diffusion cells with diclofenac, ibuprofen, flurbiprofen, ketoprofen, and naproxen and human skin. The donor solution pH significantly affected skin permeation of NSAIDs, whereas no effect of the receiver pH was observed. Similar to previous observations, the apparent permeability coefficient versus donor solution pH relationships deviated from the predictions (fractions of unionized NSAIDs) according to the acid/base theory. The influences of the viable epidermis barrier, polar pathway transport, ion permeation across skin, and effective skin pH were investigated. The effective pH values for skin permeation determined using the NSAIDs (weak acids) in this study were different from those obtained previously with a weak base at the same donor solution pH conditions, suggesting that the observed permeability-pH relationships could not be explained solely by possible pH differences between skin and donor solution. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  18. Preferential intracellular pH regulation represents a general pattern of pH homeostasis during acid-base disturbances in the armoured catfish, Pterygoplichthys pardalis.

    PubMed

    Harter, T S; Shartau, R B; Baker, D W; Jackson, D C; Val, A L; Brauner, C J

    2014-08-01

    Preferential intracellular pH (pHi) regulation, where pHi is tightly regulated in the face of a blood acidosis, has been observed in a few species of fish, but only during elevated blood PCO2. To determine whether preferential pHi regulation may represent a general pattern for acid-base regulation during other pH disturbances we challenged the armoured catfish, Pterygoplichthys pardalis, with anoxia and exhaustive exercise, to induce a metabolic acidosis, and bicarbonate injections to induce a metabolic alkalosis. Fish were terminally sampled 2-3 h following the respective treatments and extracellular blood pH, pHi of red blood cells (RBC), brain, heart, liver and white muscle, and plasma lactate and total CO2 were measured. All treatments resulted in significant changes in extracellular pH and RBC pHi that likely cover a large portion of the pH tolerance limits of this species (pH 7.15-7.86). In all tissues other than RBC, pHi remained tightly regulated and did not differ significantly from control values, with the exception of a decrease in white muscle pHi after anoxia and an increase in liver pHi following a metabolic alkalosis. Thus preferential pHi regulation appears to be a general pattern for acid-base homeostasis in the armoured catfish and may be a common response in Amazonian fishes.

  19. The Cytosolic pH of Individual Saccharomyces cerevisiae Cells Is a Key Factor in Acetic Acid Tolerance

    PubMed Central

    Fernández-Niño, Miguel; Marquina, Maribel; Swinnen, Steve; Rodríguez-Porrata, Boris

    2015-01-01

    It was shown recently that individual cells of an isogenic Saccharomyces cerevisiae population show variability in acetic acid tolerance, and this variability affects the quantitative manifestation of the trait at the population level. In the current study, we investigated whether cell-to-cell variability in acetic acid tolerance could be explained by the observed differences in the cytosolic pHs of individual cells immediately before exposure to the acid. Results obtained with cells of the strain CEN.PK113-7D in synthetic medium containing 96 mM acetic acid (pH 4.5) showed a direct correlation between the initial cytosolic pH and the cytosolic pH drop after exposure to the acid. Moreover, only cells with a low initial cytosolic pH, which experienced a less severe drop in cytosolic pH, were able to proliferate. A similar correlation between initial cytosolic pH and cytosolic pH drop was also observed in the more acid-tolerant strain MUCL 11987-9. Interestingly, a fraction of cells in the MUCL 11987-9 population showed initial cytosolic pH values below the minimal cytosolic pH detected in cells of the strain CEN.PK113-7D; consequently, these cells experienced less severe drops in cytosolic pH. Although this might explain in part the difference between the two strains with regard to the number of cells that resumed proliferation, it was observed that all cells from strain MUCL 11987-9 were able to proliferate, independently of their initial cytosolic pH. Therefore, other factors must also be involved in the greater ability of MUCL 11987-9 cells to endure strong drops in cytosolic pH. PMID:26341199

  20. Effects of acidic pH on voltage-gated ion channels in rat trigeminal mesencephalic nucleus neurons

    PubMed Central

    Han, Jin-Eon; Cho, Jin-Hwa; Choi, In-Sun; Kim, Do-Yeon

    2017-01-01

    The effects of acidic pH on several voltage-dependent ion channels, such as voltage-dependent K+ and Ca2+ channels, and hyperpolarization-gated and cyclic nucleotide-activated cation (HCN) channels, were examined using a whole-cell patch clamp technique on mechanically isolated rat mesencephalic trigeminal nucleus neurons. The application of a pH 6.5 solution had no effect on the peak amplitude of voltage-dependent K+ currents. A pH 6.0 solution slightly, but significantly inhibited the peak amplitude of voltage-dependent K+ currents. The pH 6.0 also shifted both the current-voltage and conductance-voltage relationships to the depolarization range. The application of a pH 6.5 solution scarcely affected the peak amplitude of membrane currents mediated by HCN channels, which were profoundly inhibited by the general HCN channel blocker Cs+ (1 mM). However, the pH 6.0 solution slightly, but significantly inhibited the peak amplitude of HCN-mediated currents. Although the pH 6.0 solution showed complex modulation of the current-voltage and conductance-voltage relationships, the midpoint voltages for the activation of HCN channels were not changed by acidic pH. On the other hand, voltage-dependent Ca2+ channels were significantly inhibited by an acidic pH. The application of an acidic pH solution significantly shifted the current-voltage and conductance-voltage relationships to the depolarization range. The modulation of several voltage-dependent ion channels by an acidic pH might affect the excitability of mesencephalic trigeminal nucleus neurons, and thus physiological functions mediated by the mesencephalic trigeminal nucleus could be affected in acidic pH conditions. PMID:28280415

  1. The Cytosolic pH of Individual Saccharomyces cerevisiae Cells Is a Key Factor in Acetic Acid Tolerance.

    PubMed

    Fernández-Niño, Miguel; Marquina, Maribel; Swinnen, Steve; Rodríguez-Porrata, Boris; Nevoigt, Elke; Ariño, Joaquín

    2015-11-01

    It was shown recently that individual cells of an isogenic Saccharomyces cerevisiae population show variability in acetic acid tolerance, and this variability affects the quantitative manifestation of the trait at the population level. In the current study, we investigated whether cell-to-cell variability in acetic acid tolerance could be explained by the observed differences in the cytosolic pHs of individual cells immediately before exposure to the acid. Results obtained with cells of the strain CEN.PK113-7D in synthetic medium containing 96 mM acetic acid (pH 4.5) showed a direct correlation between the initial cytosolic pH and the cytosolic pH drop after exposure to the acid. Moreover, only cells with a low initial cytosolic pH, which experienced a less severe drop in cytosolic pH, were able to proliferate. A similar correlation between initial cytosolic pH and cytosolic pH drop was also observed in the more acid-tolerant strain MUCL 11987-9. Interestingly, a fraction of cells in the MUCL 11987-9 population showed initial cytosolic pH values below the minimal cytosolic pH detected in cells of the strain CEN.PK113-7D; consequently, these cells experienced less severe drops in cytosolic pH. Although this might explain in part the difference between the two strains with regard to the number of cells that resumed proliferation, it was observed that all cells from strain MUCL 11987-9 were able to proliferate, independently of their initial cytosolic pH. Therefore, other factors must also be involved in the greater ability of MUCL 11987-9 cells to endure strong drops in cytosolic pH.

  2. Electrofocusing of acid phosphatases from frog liver, using an immobilized pH gradient.

    PubMed

    Kubicz, A; Szalewicz, A; Fawcett, J S; Chrambach, A

    1990-02-01

    Isoelectric focusing on carrier ampholyte-containing immobilized pH gradient gels was applied (i) to gels submerged in silicone oil on a Peltier cooled apparatus, (ii) to the separation of the higher molecular weight (HMW, Mr 140,000) and the lower molecular weight (LMW, Mr 38,000) acid phosphatases (AcPases) from frog livers. (i) Electrofocusing was conducted on gels submerged under silicone oil cooled and stirred on a Peltier-thermoregulated horizontal gel support plate. This procedure aimed at a) improving the temperature control of the gel by direct contact of coolant with the gel surface, and thus at being able to focus at the maximal field strength and consequently highest resolution; b) preventing evaporation from the gel and c) excluding atmospheric carbon dioxide. Silicone oil submersion did not abolish water loss from the gel into the electrolyte strips during isoelectric focusing, or a rippled gel surface. Absence of water exudation on the ripples noted previously by Atland [1] was observed. (ii) The electrofocusing of AcPases on immobilized pH gradients yielded patterns which remained stationary as a function of time, by contrast to previous analyses on carrier ampholyte generated pH gradients. The total number of enzymatically active components found in the enzyme preparations from different stages of purification and in the isolated HMW and LMW AcPases was 18. The HMW and LMW AcPases focused in characteristic pH ranges and exhibited qualitative and quantitative pattern differences. Their band patterns add up to that of a crude preparation containing both enzymes. Neither polyacrylamide gel electrophoresis (PAGE) at any nondenaturing pH, nor isoelectric focusing in carrier ampholytes with pattern changes due to the pH gradient drift were able to yield that result.

  3. Sensitivity of greenback cutthroat trout to acidic pH and elevated aluminum

    SciTech Connect

    Woodward, D.F. ); Farag, A.M. ); Little E.E.; Steadman, B. ); Yancik, R. )

    1991-01-01

    The greenback cutthroat trout Oncorhynchus clarki stomias is a threatened subspecies native to the upper South Platte and Arkansas rivers between Denver and Fort Collins, Colorado, an area also susceptible to acid deposition. In laboratory studies, the authors exposed this subspecies to nominal pHs of 4.5-6.5 and to nominal aluminum concentrations of 0, 50, 100, and 300 {mu}g/L; the control was pH 6.5 treatment without Al. The authors used soft water that contained 1.3 mg Ca/L. Exposures of 7 days each were made for four early life stages: fertilized egg, eyed embryo, alevin, and swim-up larva. Effects were measured at the end of exposure and again after a recovery period lasting until 40 days posthatch. The alevin stage was the most sensitive: at pH 5.0 with no Al, survival was reduced by 68% and swimming duration by 76%, at pH 6.0 and 50 {mu}g Al/L, swimming duration was reduced by 62%, but survival was not affected. Reductions in whole-body concentrations of Na, K, and Ca indicated organism stress. Sodium was reduced most-about 50% in alevins exposed to pH 5.0 without Al and to pH 6.0 with 50 {mu}g Al/L. Growth and the ratio of RNA to DNA were not affected by any exposure. All responses that were affected during exposure returned to normal by 40 days posthatch. Overall, it appeared that pH 6.0 and 50 {mu}g Al/L might be detrimental to greenback cutthroat trout populations.

  4. Tetracycline adsorption on kaolinite: pH, metal cations and humic acid effects.

    PubMed

    Zhao, Yanping; Geng, Jinju; Wang, Xiaorong; Gu, Xueyuan; Gao, Shixiang

    2011-07-01

    Contamination of environmental matrixes by human and animal wastes containing antibiotics is a growing health concern. Because tetracycline is one of the most widely-used antibiotics in the world, it is important to understand the factors that influence its mobility in soils. This study investigated the effects of pH, background electrolyte cations (Li(+), Na(+), K(+), Ca(2+) and Mg(2+)), heavy metal Cu(2+) and humic acid (HA) on tetracycline adsorption onto kaolinite. Results showed that tetracycline was greatly adsorbed by kaolinite over pH 3-6, then decreased with the increase of pH, indicating that tetracycline adsorption mainly through ion exchange of cations species and complexation of zwitterions species. In the presence of five types of cations (Li(+), Na(+), K(+), Ca(2+) and Mg(2+)), tetracycline adsorption decreased in accordance with the increasing of atomic radius and valence of metal cations, which suggested that outer-sphere complexes formed between tetracycline and kaolinite, and the existence of competitor ions lead to the decreasing adsorption. The presence of Cu(2+) greatly enhanced the adsorption probably by acting as a bridge ion between tetracycline species and the edge sites of kaolinite. HA also showed a major effect on the adsorption: at pH < 6, the presence of HA increased the adsorption, while the addition of HA showed little effect on tetracycline adsorption at higher pH. The soil environmental conditions, like pH, metal cations and soil organic matter, strongly influence the adsorption behavior of tetracycline onto kaolinite and need to be considered when assessing the environmental toxicity of tetracycline.

  5. Improved volatile fatty acids anaerobic production from waste activated sludge by pH regulation: Alkaline or neutral pH?

    PubMed

    Ma, Huijun; Chen, Xingchun; Liu, He; Liu, Hongbo; Fu, Bo

    2016-02-01

    In this study, the anaerobic fermentation was carried out for volatile fatty acids (VFAs) production at different pH (between 7.0 and 10.0) conditions with untreated sludge and heat-alkaline pretreated waste activated sludge. In the fermentation with untreated sludge, the extent of hydrolysis of organic matters and extent of acidification at alkaline pH are 54.37% and 30.37%, respectively, resulting in the highest VFAs yield at 235.46mg COD/gVS of three pH conditions. In the fermentation with heat-alkaline pretreated sludge, the acidification rate and VFAs yield at neutral pH are 30.98% and 240.14mg COD/gVS, respectively, which are higher than that at other pH conditions. With the glucose or bovine serum albumin as substrate for VFAs production, the neutral pH showed a higher VFAs concentration than the alkaline pH condition. The results of terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that the alkaline pH caused low microbial richness. Based on the results in this study, we demonstrated that the alkaline pH is favor of hydrolysis of organic matter in sludge while neutral pH improved the acidogenesis for the VFAs production from sludge. Our finding is obvious different to the previous research and helpful for the understanding of how heat-alkaline pretreatment and alkaline fermentation influence the VFAs production, and beneficial to the development of VFAs production process.

  6. Effect of pH on Penetration of Naphthaleneacetic Acid and Naphthaleneacetamide Through Isolated Pear Leaf Cuticle 1

    PubMed Central

    Norris, Robert F.; Bukovac, Martin J.

    1972-01-01

    Penetration of naphthaleneacetic acid through enzymatically isolated upper pear (Pyrus communis L. cv. Bartlett) leaf cuticle increased as the donor pH was decreased. Naphthaleneacetamide penetration was not influenced by donor pH. The effect of pH on naphthaleneacetic acid penetration was reversible. Higher receiver (simulated leaf interior) pH favored penetration of naphthaleneacetic acid. Changes in the degree of dissociation, and hence polarity, as controlled by hydrogen ion concentration was the prime factor in the response of naphthaleneacetic acid to pH. At pH values lower than the pK (4.2 for naphthaleneacetic acid), the molecule was primarily undissociated, lipophilic, and penetrated into the cuticle; whereas, at pH values above the pK naphthaleneacetic acid was ionized, hydrophilic, and penetrated the cuticle with difficulty or not at all. Data presented are consistent with the hypothesis that naphthaleneacetic acid and naphthaleneacetamide penetration through the cuticle takes place by diffusion. PMID:16658011

  7. Effect of pH and lactic or acetic acid on ethanol productivity by Saccharomyces cerevisiae in corn mash.

    PubMed

    Graves, Tara; Narendranath, Neelakantam V; Dawson, Karl; Power, Ronan

    2006-06-01

    The effects of lactic and acetic acids on ethanol production by Saccharomyces cerevisiae in corn mash, as influenced by pH and dissolved solids concentration, were examined. The lactic and acetic acid concentrations utilized were 0, 0.5, 1.0, 2.0, 3.0 and 4.0% w/v, and 0, 0.1, 0.2, 0.4, 0.8 and 1.6% w/v, respectively. Corn mashes (20, 25 and 30% dry solids) were adjusted to the following pH levels after lactic or acetic acid addition: 4.0, 4.5, 5.0 or 5.5 prior to yeast inoculation. Lactic acid did not completely inhibit ethanol production by the yeast. However, lactic acid at 4% w/v decreased (P<0.05) final ethanol concentration in all mashes at all pH levels. In 30% solids mash set at pH < or =5, lactic acid at 3% w/v reduced (P<0.05) ethanol production. In contrast, inhibition by acetic acid increased as the concentration of solids in the mash increased and the pH of the medium declined. Ethanol production was completely inhibited in all mashes set at pH 4 in the presence of acetic acid at concentrations > or =0.8% w/v. In 30% solids mash set at pH 4, final ethanol levels decreased (P<0.01) with only 0.1% w/v acetic acid. These results suggest that the inhibitory effects of lactic acid and acetic acid on ethanol production in corn mash fermentation when set at a pH of 5.0-5.5 are not as great as that reported thus far using laboratory media.

  8. Characterization of squamous esophageal cells resistant to bile acids at acidic pH: implication for Barrett's esophagus pathogenesis

    PubMed Central

    Goldman, Aaron; Chen, Hwu Dau Rw; Roesly, Heather B.; Hill, Kimberly A.; Tome, Margaret E.; Dvorak, Bohuslav; Bernstein, Harris

    2011-01-01

    Barrett's esophagus (BE) is a premalignant condition, where normal squamous epithelium is replaced by intestinal epithelium. BE is associated with an increased risk of developing esophageal adenocarcinoma (EAC). However, the BE cell of origin is not clear. We hypothesize that BE tissue originates from esophageal squamous cells, which can differentiate to columnar cells as a result of repeated exposure to gastric acid and bile acids, two components of refluxate implicated in BE pathology. To test this hypothesis, we repeatedly exposed squamous esophageal HET1A cells to 0.2 mM bile acid (BA) cocktail at pH 5.5 and developed an HET1AR-resistant cell line. These cells are able to survive and proliferate after repeated 2-h treatments with BA at pH 5.5. HET1AR cells are resistant to acidification and express markers of columnar differentiation, villin, CDX2, and cytokeratin 8/18. HET1AR cells have increased amounts of reactive oxygen species, concomitant with a decreased level and activity of manganese superoxide dismutase compared with parental cells. Furthermore, HET1AR cells express proteins and activate signaling pathways associated with inflammation, cell survival, and tumorigenesis that are thought to contribute to BE and EAC development. These include STAT3, NF-κB, epidermal growth factor receptor (EGFR), cyclooxygenase-2, interleukin-6, phosphorylated mammalian target of rapamycin (p-mTOR), and Mcl-1. The expression of prosurvival and inflammatory proteins and resistance to cell death could be partially modified by inhibition of STAT3 signaling. In summary, our study shows that long-term exposure of squamous cells to BA at acidic pH causes the cells to display the same characteristics and markers as BE. PMID:21127259

  9. Kinetic investigation of recombinant human hyaluronidase PH20 on hyaluronic acid.

    PubMed

    Fang, Shiping; Hays Putnam, Anna-Maria A; LaBarre, Michael J

    2015-07-01

    The kinetic investigation of hyaluronidases using physiologically relevant hyaluronic acid (HA or hyaluronan) substrate will provide useful and important clues to their catalytic behavior and function in vivo. We present here a simple and sensitive method for kinetic measurement of recombinant human hyaluronidase PH20 (rHuPH20) on HA substrates with sizes ranging from 90 to 752 kDa. The method is based on 2-aminobenzamide labeling of hydrolyzed HA products combined with separation by size exclusion-ultra performance liquid chromatography coupled with fluorescence detection. rHuPH20 was found to follow Michaelis-Menten kinetics during the initial reaction time. Optimal reaction rates were observed in the pH range of 4.5-5.5. The HA substrate size did not have significant effects on the initial rate of the reaction. By studying HA substrates of 215, 357, and 752 kDa, the kinetic parameters Km, Vmax, and kcat were determined to be 0.87-0.91 mg/ml, 1.66-1.74 NM s(-1), and 40.5-42.4 s(-1), respectively. This method allows for direct measurement of kinetics using physiologically relevant HA substrates and can be applied to other hyaluronidase kinetic measurements.

  10. Sorption of cyromazine on humic acid: Effects of pH, ionic strength and foreign ions.

    PubMed

    Zhao, Ling; Lin, Zhi-rong; Dong, Yuan-hua

    2014-02-01

    Cyromazine (CY) is a triazine pesticide used as an insect growth inhibitor for fly control in cattle manure, field crops, vegetables, and fruits. Sorption of CY onto humic acid (HA) may affect its environmental fate. In this study, HA was used to investigate the sorption of CY at different solution chemistry conditions (pH, ionic strength) and in the presence of foreign ions and norfloxacin. All sorption isotherms fitted well with the Freundlich and Langmuir models. The sorption reached a maximum at initial pH 4.0 over the initial pH range of 3.0-7.0, implying that the primary sorption mechanism was cation exchange interaction between CY+ species and the negatively charged functional groups of HA. Increasing Ca2+ concentration resulted in a considerable reduction in the Kd values of CY, hinting that Ca2+ had probably competed with CY+ for the cation exchange sites on the surfaces of HA. The sorption of CY on HA in different ionic media followed the order of NH4Cl ≈ KCl > K2SO4 > ZnCl2 ≈ CaCl2 at pH 5.0. Spectroscopic evidence demonstrated that the amino groups and triazine ring of CY was responsible for sorption onto HA, while the carboxyl group and the O-alkyl structure of HA participated in adsorbing CY.

  11. Isoelectric focusing of dansylated amino acids in immobilized pH gradients

    NASA Technical Reports Server (NTRS)

    Bianchi-Bosisio, Adriana; Righetti, Pier Giorgio; Egen, Ned B.; Bier, Milan

    1986-01-01

    The 21 free amino acids commonly encountered in proteins have been transformed into 'carrier ampholyte' species by reacting their primary amino groups with dansyl chloride. These derivatives can thus be focused in an immobilized pH gradient covering the pH interval 3.1 to 4.1, except for arginine, which still retains a pI of 8.8. Due to their inherent fluorescence, the dansyl derivatives are revealed in UV light, with a sensitivity of the order of 2-4 ng/sq mm. All nearest neighbors are separated except for the following couples: Asn-Gln, Gly-Thr, Val-Ile and Cys-Cys2, with a resolving power, in a Delta(pI) scale, of the order of 0.0018 pH units. Except for a few cases (notably the aromatic amino acids), the order of pI values is well correlated with the pK values of carboxyl groups, suggesting that the latter are not altered by dansylation. From the set of pK(COOH)-pI values of the different amino acids, the pK of the tertiary amino group in the dansyl label has been calculated to be 5.11 + or - 0.06. Knowing the pK of the amino-dansyl and the pI of the excess, free dansyl label (pI = 3.34), a pK of 1.57 is derived for its sulfonic acid group.

  12. Isoelectric focusing of dansylated amino acids in immobilized pH gradients

    NASA Technical Reports Server (NTRS)

    Bianchi-Bosisio, Adriana; Righetti, Pier Giorgio; Egen, Ned B.; Bier, Milan

    1986-01-01

    The 21 free amino acids commonly encountered in proteins have been transformed into 'carrier ampholyte' species by reacting their primary amino groups with dansyl chloride. These derivatives can thus be focused in an immobilized pH gradient covering the pH interval 3.1 to 4.1, except for arginine, which still retains a pI of 8.8. Due to their inherent fluorescence, the dansyl derivatives are revealed in UV light, with a sensitivity of the order of 2-4 ng/sq mm. All nearest neighbors are separated except for the following couples: Asn-Gln, Gly-Thr, Val-Ile and Cys-Cys2, with a resolving power, in a Delta(pI) scale, of the order of 0.0018 pH units. Except for a few cases (notably the aromatic amino acids), the order of pI values is well correlated with the pK values of carboxyl groups, suggesting that the latter are not altered by dansylation. From the set of pK(COOH)-pI values of the different amino acids, the pK of the tertiary amino group in the dansyl label has been calculated to be 5.11 + or - 0.06. Knowing the pK of the amino-dansyl and the pI of the excess, free dansyl label (pI = 3.34), a pK of 1.57 is derived for its sulfonic acid group.

  13. Thermodynamic Solubility Profile of Carbamazepine-Cinnamic Acid Cocrystal at Different pH.

    PubMed

    Keramatnia, Fatemeh; Shayanfar, Ali; Jouyban, Abolghasem

    2015-08-01

    Pharmaceutical cocrystal formation is a direct way to dramatically influence physicochemical properties of drug substances, especially their solubility and dissolution rate. Because of their instability in the solution, thermodynamic solubility of cocrystals could not be determined in the common way like other compounds; therefore, the thermodynamic solubility is calculated through concentration of their components in the eutectic point. The objective of this study is to investigate the effect of an ionizable coformer in cocrystal with a nonionizable drug at different pH. Carbamazepine (CBZ), a nonionizable drug with cinnamic acid (CIN), which is an acidic coformer, was selected to prepare CBZ-CIN cocrystal and its thermodynamic solubility was studied in pH range 2-7. Instead of HPLC that is a costly and time-consuming method, a chemometric-based approach, net analyte signal standard addition method, was selected for simultaneous determination of CBZ and CIN in solution. The result showed that, as pH increases, CIN ionization leads to change in CBZ-CIN cocrystal solubility and stability in solution. In addition, the results of this study indicated that there is no significant difference between intrinsic solubility of CBZ and cocrystal despite the higher ideal solubility of cocrystal. This verifies that ideal solubility is not good parameter to predict cocrystal solubility. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  14. Manufacture and Incorporation of Liposome-Entrapped Ethylenediaminetetraacetic Acid into Model Miniature Gouda-Type Cheese and Subsequent Effect on Starter Viability, pH, and Moisture Content.

    PubMed

    McAuliffe, Lisa N; Kilcawley, Kieran N; Sheehan, Jeremiah J; McSweeney, Paul L H

    2016-10-25

    Liposome-encapsulated ethylenediaminetetraacetic acid (EDTA) was incorporated into a model miniature Gouda-type cheese (20 g) in order to assess its effect on rennet gelation, starter viability, pH, and moisture content. EDTA was encapsulated within 2 different food-grade proliposome preparations, Pro-Lipo Duo and Pro-Lipo C (50% and 40% unsaturated soybean phospholipids and 50% and 60% aqueous medium, respectively), using the following high-shear technologies: Ultra-Turrax (5000 rpm), 2-stage homogenization (345 bar), or microfluidization (690 bar). Liposome size distribution was affected by the high-shear technology employed with the proportion of large vesicles (>100 nm) decreasing in the order microfluidization < 2-stage homogenization < Ultra-Turrax. All EDTA-containing liposomes were stable during 28 d refrigerated storage, with no significant (P ≤ 0.05) change in size distribution or EDTA entrapment efficiency (%EE). Liposome composition affected the entrapment of EDTA, with Pro-Lipo C having a significantly greater %EE than Pro-Lipo Duo, 63% and 54%, respectively. For this reason, Pro-Lipo C EDTA liposomes, with and without EDTA, were incorporated into model miniature Gouda-type cheese. Addition of liposome-encapsulated EDTA to milk during cheese making did not impact pH or rennet gel formation. No differences in composition or pH were evident in liposome-treated cheeses. The results of this study show that the incorporation of liposome-encapsulated EDTA into milk during cheese manufacture did not affect milk fermentation, moisture content, or pH, suggesting that this approach may be suitable for studying the effects of calcium equilibrium on the texture of brine-salted cheeses. © 2016 Institute of Food Technologists®.

  15. Efficient and stable enzyme immobilization in a block copolypeptide vesicle-templated biomimetic silica support.

    PubMed

    Lai, Jun-Kun; Chuang, Tzu-Han; Jan, Jeng-Shiung; Wang, Steven Sheng-Shih

    2010-10-01

    We report the immobilization of a model enzyme, papain, within silica matrices by combining vesiclization of poly-l-lysine-b-polyglycine block copolypeptides with following silica mineralization. Our novel strategy utilizes block polypeptide vesicles to induce the condensation of orthosilicic acid while trapping an enzyme within and between vesicles. The polypeptide mediated silica-immobilized enzyme exhibits enhanced pH and thermal stability and reusability, comparing with the free and vesicle encapsulated enzyme. The enhanced enzymatic activity in the immobilized enzyme is due to the confinement of the enzyme in the polypeptide mediated silica matrices. Kinetic analysis shows that the enzyme functionality is determined by the structure and property of silica/polypeptide matrices. The proposed novel strategy provides an alternative route for the synthesis of a broad range of functional bionanocomposites entrapped within silica nanostructures.

  16. pH-Sensitive polymeric micelle-based pH probe for detecting and imaging acidic biological environments.

    PubMed

    Lee, Young Ju; Kang, Han Chang; Hu, Jun; Nichols, Joseph W; Jeon, Yong Sun; Bae, You Han

    2012-09-10

    To overcome the limitations of monomeric pH probes for acidic tumor environments, this study designed a mixed micelle pH probe composed of polyethylene glycol (PEG)-b-poly(L-histidine) (PHis) and PEG-b-poly(L-lactic acid) (PLLA), which is well-known as an effective antitumor drug carrier. Unlike monomeric histidine and PHis derivatives, the mixed micelles can be structurally destabilized by changes in pH, leading to a better pH sensing system in nuclear magnetic resonance (NMR) techniques. The acidic pH-induced transformation of the mixed micelles allowed pH detection and pH mapping of 0.2-0.3 pH unit differences by pH-induced "on/off"-like sensing of NMR and magnetic resonance spectroscopy. The micellar pH probes sensed pH differences in nonbiological phosphate buffer and biological buffers such as cell culture medium and rat whole blood. In addition, the pH-sensing ability of the mixed micelles was not compromised by loaded doxorubicin. In conclusion, PHis-based micelles could have potential as a tool to simultaneously treat and map the pH of solid tumors in vivo.

  17. pH-Sensitive Polymeric Micelle-based pH Probe for Detecting and Imaging Acidic Biological Environments

    PubMed Central

    Lee, Young Ju; Kang, Han Chang; Hu, Jun; Nichols, Joseph W.; Jeon, Yong Sun; Bae, You Han

    2012-01-01

    To overcome the limitations of monomeric pH probes for acidic tumor environments, this study designed a mixed micelle pH probe composed of polyethylene glycol (PEG)-b- poly(L-histidine) (PHis) and PEG-b-poly(L-lactic acid) (PLLA), which is well-known as an effective antitumor drug carrier. Unlike monomeric histidine and PHis derivatives, the mixed micelles can be structurally destabilized by changes in pH, leading to a better pH sensing system in nuclear magnetic resonance (NMR) techniques. The acidic pH-induced transformation of the mixed micelles allowed pH detection and pH mapping of 0.2–0.3 pH unit differences by pH-induced “on/off”-like sensing of NMR and magnetic resonance spectroscopy (MRS). The micellar pH probes sensed pH differences in non-biological phosphate buffer and biological buffers such as cell culture medium and rat whole blood. In addition, the pH-sensing ability of the mixed micelles was not compromised by loaded doxorubicin. In conclusion, PHis-based micelles could have potential as a tool to simultaneously treat and map the pH of solid tumors in vivo. PMID:22861824

  18. A novel "off-on" colorimetric and fluorescent rhodamine-based pH chemosensor for extreme acidity

    NASA Astrophysics Data System (ADS)

    Tan, Jia-Lian; Zhang, Mu-Xue; Zhang, Fang; Yang, Ting-Ting; Liu, Yu; Li, Zhu-Bo; Zuo, Hua

    2015-04-01

    A novel "off-on" colorimetric and fluorescent rhodamine analogue was synthesized and characterized, and used to monitor extreme acidity (below pH 3.5) via the photophysical response to pH. The colorless spirocyclic structure at high pH (pH ⩾ 7.0) opened to the colored and highly fluorescent form at very low pH (pH < 3.0). This sensitive pH probe was characterized with short response time, good reversibility and no interaction with interfering metal ions, and the quantitative relationship between the fluorescence intensity and pH value was consistent with the equilibrium equation pH = pKa - log[(Imax - I)/(I - Imin)]. The fluorescent response to strong acidity was further verified by fluorescent imaging of bacteria, Escherichia coli, which contributed to the development of more useful colorimetric and fluorescent sensors based on the rhodamine platform for measuring intracellular pH in extremely acidic conditions.

  19. Study of metabolic profile of Rhizopus oryzae to enhance fumaric acid production under low pH condition.

    PubMed

    Liu, Ying; Xu, Qing; Lv, Chunwei; Yan, Caixia; Li, Shuang; Jiang, Ling; Huang, He; Ouyang, Pingkai

    2015-12-01

    Ensuring a suitable pH is a major problem in industrial organic acid fermentation. To circumvent this problem, we used a metabolic profiling approach to analyze metabolite changes in Rhizopus oryzae under different pH conditions. A correlation between fumaric acid production and intracellular metabolic characteristics of R. oryzae was revealed by principal component analysis. The results showed that to help cell survival in the presence of low pH, R. oryzae altered amino acid and fatty acid metabolism and promoted sugar or sugar alcohol synthesis, corresponding with a suppressing of energy metabolism, phenylalanine, and tyrosine synthesis and finally resulting in the low performance of fumaric acid production. Based on this observation, 1 % linoleic acid was added to the culture medium in pH 3.0 to decrease the carbon demand for cell survival, and the fumaric acid titer was enhanced by 39.7 % compared with the control (pH 3.0 without linoleic acid addition), reaching 18.3 g/L after 84 h of fermentation. These findings provide new insights into the mechanism by which R. oryzae responds to acidic stress and would be helpful for the development of efficient strategies for fumaric acid production at low pH.

  20. Generation of Gastroesophageal Reflux Disease Symptoms During Esophageal Acid Infusion With Concomitant Esophageal pH Monitoring in Healthy Adults.

    PubMed

    Ohara, Shunji; Furuta, Kenji; Adachi, Kyoichi; Fukazawa, Kousuke; Aimi, Masahito; Miki, Masaharu; Kinoshita, Yoshikazu

    2013-10-01

    The sensitivity of the upper and lower esophageal mucosa to acid is considered to differ. We investigated the relationship between pH changes in different sites of the esophagus and generation of gastroesophageal reflux symptoms during an acid infusion test. An acid infusion catheter was placed at 5 or 15 cm above the lower esophageal sphincter (LES) in 18 healthy volunteers, while a 2-channel pH sensor catheter was also placed in each with the sensors set at 5 and 15 cm above the LES. Solutions containing water and hydrochloric acid at different concentrations were infused through the infusion catheter. Acid infusion in the upper esophagus caused a pH drop in both upper and lower esophageal sites, whereas that in the lower esophagus resulted in a significant pH drop only in the lower without a corresponding pH decline in the upper esophagus. Stronger heartburn, chest pain, and chest oppression symptoms were noted when acid was infused in the upper as compared to the lower esophagus, while increased intra-esophageal acidity strengthened each symptom. Regurgitations caused by upper and lower esophageal acid infusions were similar, and not worsened by a larger drop in intra-esophageal pH. Chest pain was caused only by lowered intra-esophageal pH, while heartburn, chest oppression, and regurgitation were induced by a less acidic solution. Higher intra-esophageal acidity caused stronger heartburn, chest pain, and chest oppression symptoms. However, regurgitation was not significantly influenced by intra-esophageal acidity. The upper esophagus showed higher acid sensitivity than the lower esophagus.

  1. Biogenic precipitation of manganese oxides and enrichment of heavy metals at acidic soil pH

    NASA Astrophysics Data System (ADS)

    Mayanna, Sathish; Peacock, Caroline L.; Schäffner, Franziska; Grawunder, Anja; Merten, Dirk; Kothe, Erika; Büchel, Georg

    2014-05-01

    The precipitation of biogenic Mn oxides at acidic pH is rarely reported and poorly understood, compared to biogenic Mn oxide precipitation at near neutral conditions. Here we identified and investigated the precipitation of biogenic Mn oxides in acidic soil, and studied their role in the retention of heavy metals, at the former uranium mining site of Ronneburg, Germany. The site is characterized by acidic pH, low carbon content and high heavy metal loads including rare earth elements. Specifically, the Mn oxides were present in layers identified by detailed soil profiling and within these layers pH varied from 4.7 to 5.1, Eh varied from 640 to 660 mV and there were enriched total metal contents for Ba, Ni, Co, Cd and Zn in addition to high Mn levels. Using electron microprobe analysis, synchrotron X-ray diffraction and X-ray absorption spectroscopy, we identified poorly crystalline birnessite (δ-MnO2) as the dominant Mn oxide in the Mn layers, present as coatings covering and cementing quartz grains. With geochemical modelling we found that the environmental conditions at the site were not favourable for chemical oxidation of Mn(II), and thus we performed 16S rDNA sequencing to isolate the bacterial strains present in the Mn layers. Bacterial phyla present in the Mn layers belonged to Firmicutes, Actinobacteria and Proteobacteria, and from these phyla we isolated six strains of Mn(II) oxidizing bacteria and confirmed their ability to oxidise Mn(II) in the laboratory. The biogenic Mn oxide layers act as a sink for metals and the bioavailability of these metals was much lower in the Mn layers than in adjacent layers, reflecting their preferential sorption to the biogenic Mn oxide. In this presentation we will report our findings, concluding that the formation of natural biogenic poorly crystalline birnessite can occur at acidic pH, resulting in the formation of a biogeochemical barrier which, in turn, can control the mobility and bioavailability of heavy metals in

  2. pH dependence of the properties of waterborne pressure-sensitive adhesives containing acrylic acid.

    PubMed

    Wang, Tao; Canetta, Elisabetta; Weerakkody, Tecla G; Keddie, Joseph L; Rivas, Urko

    2009-03-01

    Polymer colloids are often copolymerized with acrylic acid monomers in order to impart colloidal stability. Here, the effects of the pH on the nanoscale and macroscopic adhesive properties of waterborne poly(butyl acrylate-co-acrylic acid) films are reported. In films cast from acidic colloidal dispersions, hydrogen bonding between carboxylic acid groups dominates the particle-particle interactions, whereas ionic dipolar interactions are dominant in films cast from basic dispersions. Force spectroscopy using an atomic force microscope and macroscale mechanical measurements show that latex films with hydrogen-bonding interactions have lower elastic moduli and are more deformable. They yield higher adhesion energies. On the other hand, in basic latex, ionic dipolar interactions increase the moduli of the dried films. These materials are stiffer and less deformable and, consequently, exhibit lower adhesion energies. The rate of water loss from acidic latex is slower, perhaps because of hydrogen bonding with the water. Therefore, although acid latex offers greater adhesion, there is a limitation in the film formation.

  3. Influence of polysorbate 80 and cyclopropane fatty acid synthase activity on lactic acid production by Lactobacillus casei ATCC 334 at low pH.

    PubMed

    Broadbent, J R; Oberg, T S; Hughes, J E; Ward, R E; Brighton, C; Welker, D L; Steele, J L

    2014-03-01

    Lactic acid is an important industrial chemical commonly produced through microbial fermentation. The efficiency of acid extraction is increased at or below the acid's pKa (pH 3.86), so there is interest in factors that allow for a reduced fermentation pH. We explored the role of cyclopropane synthase (Cfa) and polysorbate (Tween) 80 on acid production and membrane lipid composition in Lactobacillus casei ATCC 334 at low pH. Cells from wild-type and an ATCC 334 cfa knockout mutant were incubated in APT broth medium containing 3 % glucose plus 0.02 or 0.2 % Tween 80. The cultures were allowed to acidify the medium until it reached a target pH (4.5, 4.0, or 3.8), and then the pH was maintained by automatic addition of NH₄OH. Cells were collected at the midpoint of the fermentation for membrane lipid analysis, and media samples were analyzed for lactic and acetic acids when acid production had ceased. There were no significant differences in the quantity of lactic acid produced at different pH values by wild-type or mutant cells grown in APT, but the rate of acid production was reduced as pH declined. APT supplementation with 0.2 % Tween 80 significantly increased the amount of lactic acid produced by wild-type cells at pH 3.8, and the rate of acid production was modestly improved. This effect was not observed with the cfa mutant, which indicated Cfa activity and Tween 80 supplementation were each involved in the significant increase in lactic acid yield observed with wild-type L. casei at pH 3.8.

  4. Sulfate reduction at low pH to remediate acid mine drainage.

    PubMed

    Sánchez-Andrea, Irene; Sanz, Jose Luis; Bijmans, Martijn F M; Stams, Alfons J M

    2014-03-30

    Industrial activities and the natural oxidation of metallic sulfide-ores produce sulfate-rich waters with low pH and high heavy metals content, generally termed acid mine drainage (AMD). This is of great environmental concern as some heavy metals are highly toxic. Within a number of possibilities, biological treatment applying sulfate-reducing bacteria (SRB) is an attractive option to treat AMD and to recover metals. The process produces alkalinity, neutralizing the AMD simultaneously. The sulfide that is produced reacts with the metal in solution and precipitates them as metal sulfides. Here, important factors for biotechnological application of SRB such as the inocula, the pH of the process, the substrates and the reactor design are discussed. Microbial communities of sulfidogenic reactors treating AMD which comprise fermentative-, acetogenic- and SRB as well as methanogenic archaea are reviewed. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Influence of pH, bleaching agents, and acid etching on surface wear of bovine enamel.

    PubMed

    Soares, Ana Flávia; Bombonatti, Juliana Fraga Soares; Alencar, Marina Studart; Consolmagno, Elaine Cristina; Honório, Heitor Marques; Mondelli, Rafael Francisco Lia

    2016-01-01

    Development of new materials for tooth bleaching justifies the need for studies to evaluate the changes in the enamel surface caused by different bleaching protocols. The aim of this study was to evaluate the bovine dental enamel wear in function of different bleaching gel protocols, acid etching and pH variation. Sixty fragments of bovine teeth were cut, obtaining a control and test areas. In the test area, one half received etching followed by a bleaching gel application, and the other half, only the bleaching gel. The fragments were randomly divided into six groups (n=10), each one received one bleaching session with five hydrogen peroxide gel applications of 8 min, activated with hybrid light, diode laser/blue LED (HL) or diode laser/violet LED (VHL) (experimental): Control (C); 35% Total Blanc Office (TBO35HL); 35% Lase Peroxide Sensy (LPS35HL); 25% Lase Peroxide Sensy II (LPS25HL); 15% Lase Peroxide Lite (LPL15HL); and 10% hydrogen peroxide (experimental) (EXP10VHL). pH values were determined by a pHmeter at the initial and final time periods. Specimens were stored, subjected to simulated brushing cycles, and the superficial wear was determined (μm). ANOVA and Tukey´s tests were applied (α=0.05). The pH showed a slight decrease, except for Group LPL15HL. Group LPS25HL showed the highest degree of wear, with and without etching. There was a decrease from the initial to the final pH. Different bleaching gels were able to increase the surface wear values after simulated brushing. Acid etching before bleaching increased surface wear values in all groups.

  6. Influence of pH, bleaching agents, and acid etching on surface wear of bovine enamel

    PubMed Central

    Soares, Ana Flávia; Bombonatti, Juliana Fraga Soares; Alencar, Marina Studart; Consolmagno, Elaine Cristina; Honório, Heitor Marques; Mondelli, Rafael Francisco Lia

    2016-01-01

    ABSTRACT Development of new materials for tooth bleaching justifies the need for studies to evaluate the changes in the enamel surface caused by different bleaching protocols. Objective The aim of this study was to evaluate the bovine dental enamel wear in function of different bleaching gel protocols, acid etching and pH variation. Material and Methods Sixty fragments of bovine teeth were cut, obtaining a control and test areas. In the test area, one half received etching followed by a bleaching gel application, and the other half, only the bleaching gel. The fragments were randomly divided into six groups (n=10), each one received one bleaching session with five hydrogen peroxide gel applications of 8 min, activated with hybrid light, diode laser/blue LED (HL) or diode laser/violet LED (VHL) (experimental): Control (C); 35% Total Blanc Office (TBO35HL); 35% Lase Peroxide Sensy (LPS35HL); 25% Lase Peroxide Sensy II (LPS25HL); 15% Lase Peroxide Lite (LPL15HL); and 10% hydrogen peroxide (experimental) (EXP10VHL). pH values were determined by a pHmeter at the initial and final time periods. Specimens were stored, subjected to simulated brushing cycles, and the superficial wear was determined (μm). ANOVA and Tukey´s tests were applied (α=0.05). Results The pH showed a slight decrease, except for Group LPL15HL. Group LPS25HL showed the highest degree of wear, with and without etching. Conclusion There was a decrease from the initial to the final pH. Different bleaching gels were able to increase the surface wear values after simulated brushing. Acid etching before bleaching increased surface wear values in all groups. PMID:27008254

  7. Mycorrhizal Response to Experimental pH and P Manipulation in Acidic Hardwood Forests

    PubMed Central

    Kluber, Laurel A.; Carrino-Kyker, Sarah R.; Coyle, Kaitlin P.; DeForest, Jared L.; Hewins, Charlotte R.; Shaw, Alanna N.; Smemo, Kurt A.; Burke, David J.

    2012-01-01

    Many temperate forests of the Northeastern United States and Europe have received significant anthropogenic acid and nitrogen (N) deposition over the last century. Although temperate hardwood forests are generally thought to be N-limited, anthropogenic deposition increases the possibility of phosphorus (P) limiting productivity in these forest ecosystems. Moreover, inorganic P availability is largely controlled by soil pH and biogeochemical theory suggests that forests with acidic soils (i.e., <pH 5) are particularly vulnerable to P limitation. Results from previous studies in these systems are mixed with evidence both for and against P limitation. We hypothesized that shifts in mycorrhizal colonization and community structure help temperate forest ecosystems overcome an underlying P limitation by accessing mineral and organic P sources that are otherwise unavailable for direct plant uptake. We examined arbuscular mycorrhizal (AM) and ectomycorrhizal (EcM) communities and soil microbial activity in an ecosystem-level experiment where soil pH and P availability were manipulated in mixed deciduous forests across eastern Ohio, USA. One year after treatment initiation, AM root biomass was positively correlated with the most available P pool, resin P, while AM colonization was negatively correlated. In total, 15,876 EcM root tips were identified and assigned to 26 genera and 219 operational taxonomic units (97% similarity). Ectomycorrhizal richness and root tip abundance were negatively correlated with the moderately available P pools, while the relative percent of tips colonized by Ascomycetes was positively correlated with soil pH. Canonical correspondence analysis revealed regional, but not treatment, differences in AM communities, while EcM communities had both treatment and regional differences. Our findings highlight the complex interactions between mycorrhizae and the soil environment and further underscore the fact that mycorrhizal communities do not merely

  8. Mycorrhizal response to experimental pH and P manipulation in acidic hardwood forests.

    PubMed

    Kluber, Laurel A; Carrino-Kyker, Sarah R; Coyle, Kaitlin P; DeForest, Jared L; Hewins, Charlotte R; Shaw, Alanna N; Smemo, Kurt A; Burke, David J

    2012-01-01

    Many temperate forests of the Northeastern United States and Europe have received significant anthropogenic acid and nitrogen (N) deposition over the last century. Although temperate hardwood forests are generally thought to be N-limited, anthropogenic deposition increases the possibility of phosphorus (P) limiting productivity in these forest ecosystems. Moreover, inorganic P availability is largely controlled by soil pH and biogeochemical theory suggests that forests with acidic soils (i.e., <pH 5) are particularly vulnerable to P limitation. Results from previous studies in these systems are mixed with evidence both for and against P limitation. We hypothesized that shifts in mycorrhizal colonization and community structure help temperate forest ecosystems overcome an underlying P limitation by accessing mineral and organic P sources that are otherwise unavailable for direct plant uptake. We examined arbuscular mycorrhizal (AM) and ectomycorrhizal (EcM) communities and soil microbial activity in an ecosystem-level experiment where soil pH and P availability were manipulated in mixed deciduous forests across eastern Ohio, USA. One year after treatment initiation, AM root biomass was positively correlated with the most available P pool, resin P, while AM colonization was negatively correlated. In total, 15,876 EcM root tips were identified and assigned to 26 genera and 219 operational taxonomic units (97% similarity). Ectomycorrhizal richness and root tip abundance were negatively correlated with the moderately available P pools, while the relative percent of tips colonized by Ascomycetes was positively correlated with soil pH. Canonical correspondence analysis revealed regional, but not treatment, differences in AM communities, while EcM communities had both treatment and regional differences. Our findings highlight the complex interactions between mycorrhizae and the soil environment and further underscore the fact that mycorrhizal communities do not merely

  9. Intracellular pH Response to Weak Acid Stress in Individual Vegetative Bacillus subtilis Cells.

    PubMed

    Pandey, Rachna; Vischer, Norbert O E; Smelt, Jan P P M; van Beilen, Johan W A; Ter Beek, Alexander; De Vos, Winnok H; Brul, Stanley; Manders, Erik M M

    2016-11-01

    Intracellular pH (pHi) critically affects bacterial cell physiology. Hence, a variety of food preservation strategies are aimed at perturbing pHi homeostasis. Unfortunately, accurate pHi quantification with existing methods is suboptimal, since measurements are averages across populations of cells, not taking into account interindividual heterogeneity. Yet, physiological heterogeneity in isogenic populations is well known to be responsible for differences in growth and division kinetics of cells in response to external stressors. To assess in this context the behavior of intracellular acidity, we have developed a robust method to quantify pHi at single-cell levels in Bacillus subtilis Bacilli spoil food, cause disease, and are well known for their ability to form highly stress-resistant spores. Using an improved version of the genetically encoded ratiometric pHluorin (IpHluorin), we have quantified pHi in individual B. subtilis cells, cultured at an external pH of 6.4, in the absence or presence of weak acid stresses. In the presence of 3 mM potassium sorbate, a decrease in pHi and an increase in the generation time of growing cells were observed. Similar effects were observed when cells were stressed with 25 mM potassium acetate. Time-resolved analysis of individual bacteria in growing colonies shows that after a transient pH decrease, long-term pH evolution is highly cell dependent. The heterogeneity at the single-cell level shows the existence of subpopulations that might be more resistant and contribute to population survival. Our approach contributes to an understanding of pHi regulation in individual bacteria and may help scrutinizing effects of existing and novel food preservation strategies.

  10. Proton conductance and intracellular pH recovery from an acid load in chicken enterocytes.

    PubMed Central

    Peral, M J; Ilundáin, A A

    1995-01-01

    1. Chicken enterocytes present a Na(+)-independent proton transport mechanism involved in pHi recovery from an acid load. In the current study the nature of this proton transport system is investigated. 2. The pHi of acid-loaded cells increased when transferred to Na(+)-free, pH 7.4 buffers, both at 6 and 65 mM extracellular potassium concentration. Addition of nigericin accelerated the rate of cell alkalinization. 3. When acid-loaded cells were transferred to a Na(+)-free, pH 6.5 buffer, the cells acidified further, regardless of the extracellular potassium concentration. The addition of nigericin increased the rate of acidification at 6 mM K+ but produced an alkalinization at 65 mM K+. 4. The rate of the Na(+)-independent regulatory cell alkalinization was inhibited by SCH 28080, DCCD, NBD-Cl, rotenone or Zn2+. Addition of valinomycin reversed the inhibition induced by SCH 28080, DCCD and NBD-Cl but not that induced by Zn2+ or rotenone. Zn2+ inhibition was abolished by the metal chelator DTPA. 5. Cytosolic acidification increased the rate of Na(+)-independent regulatory cell alkalinization. 6. The results suggest that the Na(+)-independent proton transport system is a Zn(2+)-sensitive proton-conducting pathway which is regulated by the cytosolic proton concentration. PMID:7602517

  11. Effect of substrate concentration on inhibition of prostaglandin synthetase of bull seminal vesicles by anti-inflammatory drugs and fenamic acid analogs.

    PubMed

    Cushman, D W; Cheung, H S

    1976-03-26

    Although microsomes of bull seminal vesicle synthesize prostaglandins F2alpha, E2 and D2 from arachidonic acid under suitable assay conditions, prostaglandin E2 is the only significant product at either low concentration of arachidonic acid or high concentration of microsomes. Studies of inhibition of prostaglandin synthesis in vitro by anti-inflammatory drugs at both high (1 mM) and low (1 muM) concentrations of arachidonic acid, suggest three distinct mechanisms of inhibition. Benzydamine and flazalone are non-competitive or weakly competitive with arachidonic acid and, at high concentrations of arachidonic acid, they augment sythesis of prostaglandin E2 while inhibiting production of prostaglandins F2alpha and D2. Niflumic acid and the arylacetic acids naproxen and ibuprofen are competitive inhibiting all products equally, but with 100-500-fold greater potency at the low substrate concentration. The fenamic acids, indomethacin, aspirin, and phenylbutazone also inhibit equally all prostaglandin products, but are only 20--50 times more potent at the low substrate concentration. Studies with analogs of the fenamic acids indicate that the diphenylamine protion of their structure is essential for inhibition of prostaglandin synthesis, whereas the o-carboxyl and m-alkul substitutents greatly enhance inhibitory potency.

  12. An HPLC method with UV detection, pH control, and reductive ascorbic acid for cyanuric acid analysis in water.

    PubMed

    Cantú, R; Evans, O; Kawahara, F K; Shoemaker, J A; Dufour, A P

    2000-12-01

    Every year over 250 million pounds of cyanuric acid (CA) and chlorinated isocyanurates are produced industrially. These compounds are standard ingredients in formulations for household bleaches, industrial cleansers, dishwasher compounds, general sanitizers, and chlorine stabilizers. The method developed for CA using high-performance liquid chromatography (HPLC) with UV detection simplifies and optimizes certain parameters of previous methodologies by effective pH control of the eluent (95% phosphate buffer: 5% methanol, v/v) to the narrow pH range of 7.2-7.4. UV detection was set at the optimum wavelength of 213 nm where the cyanuric ion absorbs strongly. Analysis at the lower pH range of 6.8-7.1 proved inadequate due to CA keto-enol tautomerism, while at pHs of <6.8 there were substantial losses in analytical sensitivity. In contrast, pHs of >7.4 proved more sensitive but their use was rejected because of CA elution at the chromatographic void volume and due to chemical interferences. The complex equilibria of chlorinated isocyanurates and associated species were suppressed by using reductive ascorbic acid to restrict the products to CA. UV, HPLC-UV, and electrospray ionization mass spectrometry techniques were combined to monitor the reactive chlorinated isocyanurates and to support the use of ascorbic acid. The resulting method is reproducible and measures CA in the 0.5-125 mg/L linear concentration range with a method detection limit of 0.05 mg/L in water.

  13. Effect of pH on complex formation between debranched waxy rice starch and fatty acids.

    PubMed

    Yotsawimonwat, Songwut; Sriroth, Klanarong; Kaewvichit, Sayam; Piyachomkwan, Kaukoon; Jane, Jay-Lin; Sirithunyalug, Jakkapan

    2008-08-15

    Complex formations between debranched waxy rice starch (DBS) and fatty acids (FA) of different hydrocarbon chain lengths (8:0, 10:0, 12:0, 14:0, 16:0, and 18:0) were studied in an aqueous solution by measuring the blue colour stained with iodine. The objective of this study was to understand the effects of the solubility and hydrophobicity of guest molecules (FA) on the complex formation with DBS. Lauric acid (12:0) displayed the greatest complex forming ability with DBS by showing the least blue colour developed with iodine. The effect of pH (3-7) on the DBS/FA complex formation was evaluated by measuring the iodine-scanning spectra of the mixture. Short-chain FA (8:0) displayed less complex formation at pH>or=5, above the pK(a) of fatty acid (approximately 4.8), which suggested that the charge formation of the short-chain FA caused a lower partitioning of the FA into the hydrophobic cavity of the DBS single helix. On the contrary, FA of 10:0-18:0 displayed an increased complex formation at pH>5, which could be attributed to increased solubility of these longer-chain FA at a dissociated and ionized form. The hydrocarbon chain length of the FA had an important impact on the extent of the complex formation. A FA that had a shorter hydrocarbon chain was more soluble in an aqueous solution and more readily formed a complex with DBS. At pH 6 and 7 (above the pK(a)), 10:0 formed less inclusion complexes with DBS than did 12:0. Iodine-scanning spectra showed that the absorbances of all iodine-stained DBS/FA solutions at higher wavelength were substantially lower than that of the iodine-stained DBS alone, suggesting that FA preferentially formed inclusion complexes with DBS of longer chains.

  14. Effects of pH and acid concentration on erosive dissolution of enamel, dentine, and compressed hydroxyapatite.

    PubMed

    Shellis, R P; Barbour, M E; Jones, S B; Addy, M

    2010-10-01

    The aims of this study were to determine the effects of pH and acid concentration on the dissolution of enamel, dentine, and compressed hydroxyapatite (HA) in citric acid solutions (15.6 and 52.1 mmol l(-1) ; pH 2.45, 3.2, and 3.9), using a pH-stat system. After an initial adjustment period, the dissolution rates of enamel and HA were constant, while that of dentine decreased with time. The dissolution rate increased as the pH decreased, and this was most marked for enamel. To compare substrates, the rate of mineral dissolution was normalized to the area occupied by mineral at the specimen surface. For a given acid concentration, the normalized dissolution rate of HA was always less than that for either dentine or enamel. The dissolution rate for dentine mineral was similar to that for enamel at pH 2.45 and greater at pH 3.2 and pH 3.9. The concentration of acid significantly affected the enamel dissolution rate at pH 2.45 and pH 3.2, but not at pH 3.9, and did not significantly affect the dissolution rates of dentine or HA at any pH. The variation in response of the dissolution rate to acid concentration/buffer capacity with respect to pH and tissue type might complicate attempts to predict erosive potential from solution composition.

  15. Analysis of a mixture of a known and an unknown weak acid by titration to a preset pH.

    PubMed

    Livaska, A

    1975-12-01

    The preset-pH titration method has been used to indicate the presence of a second acid when the titration curve (pH vs. volume of added titrant) seems to indicate only one acid. By use of the method even small amounts of propionic acid can be detected in an acetic acid solution despite the small value of Delta log K(H)(HA) = 0.18. Binary mixtures of acids may bs analysed when one acid is known, and log k(H)(HA) for the unknown acid may be found. Acetic acid, as the known acid, has been determined together with hydrochloric, mandelic, hydroxyacetic or boric acid or ammonium ion, with an error of about 1%. The method can be used in some cases for titration of ternary mixtures of one known and two unknown acids. Only the sum of the unknown acids can then be determined together with the known acid.

  16. Effect of pH, temperature, and potassium sorbate on amino acid uptake in Salmonella typhimurium 7136.

    PubMed Central

    Tuncan, E U; Martin, S E

    1985-01-01

    The effect of sorbate on L-serine and L-histidine uptake in Salmonella typhimurium was studied at various pH levels, temperatures, and amino acid and sorbate concentrations. Low pH had an apparent synergistic effect on amino acid uptake inhibition caused by sorbate. The relationship between sorbate concentration and the amount of amino acid uptake inhibition was not linear. Compared with L-histidine, L-serine uptake was more sensitive to changes in pH, temperature, and sorbate concentration. Various degrees of amino acid uptake inhibition by sorbate may be related to differences between amino acid transport systems. The results of this study suggest that sorbate acts as a noncompetitive inhibitor of amino acid uptake in S. typhimurium. PMID:3888108

  17. Interaction of alpha-lactalbumin with dimyristoyl phosphatidylcholine vesicles. II. A fluorescence polarization study.

    PubMed

    Herreman, W; van Tornout, P; van Cauwelaert, F H; Hanssens, I

    1981-01-22

    The interaction of alpha-lactalbumin with dimyristoyl phosphatidylcholine vesicles was studied as a function of temperature, pH and the molar ratio of phospholipid to protein. The method consisted of measuring the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene used as a probe embedded in the vesicles. After incubation of the protein with the phospholipid for 2 h at 23 degrees C, the polarization of the light emitted by this probe shifted to higher values; the shift was greater at acidic pH than at neutral pH. After incubation at 37 degrees C, no shift in polarization was found at pH 7, 6 and 5 while a strong increase occurred at pH 4. Lowering the temperature, after incubation at 37 degrees C, had little effect on the polarization at neutral pH. At pH 5, however, and in the transition range of the phospholipid, the polarization increased greatly. A kinetic study of the interaction carried out around the transition temperature of dimyristoyl phosphatidylcholine as a function of pH shows that the speed of complex formation between alpha-lactalbumin and the lipid increases from neutral to acidic pH. From the present results and in agreement with our earlier calorimetric and fluorescence data (Hanssens, I., Houthuys, C., Herreman, W. and van Cauwelaert, F.H. (1980) Biochim. Biophys, Acta 602, 539--557), it is concluded that at neutral pH the interaction mechanism is probably different from that at acidic pH. At neutral pH and at all temperatures, alpha-lactalbumin is mainly absorbed electrostatically to the outer surface of the vesicle with little or no influence on the transition temperature of the phospholipid. At this pH, only around the transition temperature is penetration possible. At pH 4, however, the protein is able to penetrate the vesicle at all temperatures and to interact hydrophobically with the phospholipid fatty acid chains. As a result of this interaction, the transition temperature is increased by about 4 degrees C. This different

  18. Intracellular pH regulation by acid-base transporters in mammalian neurons.

    PubMed

    Ruffin, Vernon A; Salameh, Ahlam I; Boron, Walter F; Parker, Mark D

    2014-01-01

    Intracellular pH (pHi) regulation in the brain is important in both physiological and physiopathological conditions because changes in pHi generally result in altered neuronal excitability. In this review, we will cover 4 major areas: (1) The effect of pHi on cellular processes in the brain, including channel activity and neuronal excitability. (2) pHi homeostasis and how it is determined by the balance between rates of acid loading (J L) and extrusion (J E). The balance between J E and J L determine steady-state pHi, as well as the ability of the cell to defend pHi in the face of extracellular acid-base disturbances (e.g., metabolic acidosis). (3) The properties and importance of members of the SLC4 and SLC9 families of acid-base transporters expressed in the brain that contribute to J L (namely the Cl-HCO3 exchanger AE3) and J E (the Na-H exchangers NHE1, NHE3, and NHE5 as well as the Na(+)- coupled HCO3 (-) transporters NBCe1, NBCn1, NDCBE, and NBCn2). (4) The effect of acid-base disturbances on neuronal function and the roles of acid-base transporters in defending neuronal pHi under physiopathologic conditions.

  19. Intracellular pH regulation by acid-base transporters in mammalian neurons

    PubMed Central

    Ruffin, Vernon A.; Salameh, Ahlam I.; Boron, Walter F.; Parker, Mark D.

    2014-01-01

    Intracellular pH (pHi) regulation in the brain is important in both physiological and physiopathological conditions because changes in pHi generally result in altered neuronal excitability. In this review, we will cover 4 major areas: (1) The effect of pHi on cellular processes in the brain, including channel activity and neuronal excitability. (2) pHi homeostasis and how it is determined by the balance between rates of acid loading (JL) and extrusion (JE). The balance between JE and JL determine steady-state pHi, as well as the ability of the cell to defend pHi in the face of extracellular acid-base disturbances (e.g., metabolic acidosis). (3) The properties and importance of members of the SLC4 and SLC9 families of acid-base transporters expressed in the brain that contribute to JL (namely the Cl-HCO3 exchanger AE3) and JE (the Na-H exchangers NHE1, NHE3, and NHE5 as well as the Na+- coupled HCO3− transporters NBCe1, NBCn1, NDCBE, and NBCn2). (4) The effect of acid-base disturbances on neuronal function and the roles of acid-base transporters in defending neuronal pHi under physiopathologic conditions. PMID:24592239

  20. Influence of five neutralizing products on intra-oral pH after rinsing with simulated gastric acid.

    PubMed

    Lindquist, Birgitta; Lingström, Peter; Fändriks, Lars; Birkhed, Dowen

    2011-08-01

    The aetiology of dental erosion may be of both extrinsic and intrinsic origin. The aim of the present study was to test the ability of various neutralizing products to raise the low intra-oral pH after an erosive exposure, in this case to gastric acid, which was simulated using hydrochloric acid (HCl). Eleven adults participated. They rinsed with 10 ml of 10 mM HCl (pH 2) or 10 ml of 100 mM HCl (pH 1) for 1 min, after which the pH was measured intra-orally for up to 30 min at four sites (two approximal, one buccal, and the dorsum of the tongue). After rinsing with the two acid solutions (pH 1 and pH 2), the following products were used: (i) antacid tablet; (ii) gum arabic lozenge; (iii) mineral water; (iv) milk; and (v) tap water (positive control). The negative control was no product use. The five test products were used for 2 min after the erosive challenge. All the products produced an initially higher pH compared with the negative control. The antacid tablet resulted in the greatest and most rapid increase in pH, followed by the lozenge. In dental practice, the use of any of the neutralizing products tested, especially the antacid tablet, could be recommended in order to increase the intra-oral pH after an erosive challenge. © 2011 Eur J Oral Sci.

  1. External concentration of organic acid anions and pH: key independent variables for studying how organic acids inhibit growth of bacteria in mildly acidic foods.

    PubMed

    Carpenter, C E; Broadbent, J R

    2009-01-01

    Although the mechanisms by which organic acids inhibit growth of bacteria in mildly acidic foods are not fully understood, it is clear that intracellular accumulation of anions is a primary contributor to inhibition of bacterial growth. We hypothesize that intracellular accumulation of anions is driven by 2 factors, external anion concentration and external acidity. This hypothesis follows from basic chemistry principles that heretofore have not been fully applied to studies in the field, and it has led us to develop a novel approach for predicting internal anion concentration by controlling the external concentration of anions and pH. This approach overcomes critical flaws in contemporary experimental design that invariably target concentration of either protonated acid or total acid in the growth media thereby leaving anion concentration to vary depending on the pK(a) of the acids involved. Failure to control external concentration of anions has undoubtedly confounded results, and it has likely led to misleading conclusions regarding the antimicrobial action of organic acids. In summary, we advocate an approach for directing internal anion levels by controlling external concentration of anions and pH because it presents an additional opportunity to study the mechanisms by which organic acids inhibit bacterial growth. Knowledge gained from such studies would have important application in the control of important foodborne pathogens such as Listeria monocytogenes, and may also facilitate efforts to promote the survival in foods or beverages of desirable probiotic bacteria.

  2. The role of low molecular weight organic acids on controlling pH in coastal sea water

    NASA Astrophysics Data System (ADS)

    Ding, H.

    2015-12-01

    Series investigation of the Jiaozhou Bay, China, observed existences of three low molecular weight organic acids (LMWOAs), including lactic acid, acetic acid and formic acid, with high concentration in the sea water. Generally, their amount accounted for about 20% of DOC in the sea water of the bay. Human activities around the bay were considered as the major source of the LMWOAs. Also, long term detection showed that the pH value in the Jiaozhou Bay was lower than that in the adjacent Yellow Sea. On average, the difference of pH values between the bay and the Yellow was about 0.2. Due to higher concentrations of the LMWOAs, their contribution to lower pH value of the bay should not be ignored. To validate the effect of LMWOAs on the pH value of the bay, a new software was developed to calculate the pH value in the sea water samples based on alkalinity by adding three items of the three organic acids in the expression. Compared to the traditional pH calculating software, the new software could improve the calculating results significantly. Our results confirmed that LMWOAs was an important control factor to adjust pH values in coastal area.

  3. pH dependence of methyl phosphonic acid, dipicolinic acid, and cyanide by surface-enhanced Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Farquharson, Stuart; Gift, Alan; Maksymiuk, Paul; Inscore, Frank E.; Smith, Wayne W.

    2004-03-01

    U.S. and Coalition forces fighting terrorism in Afghanistan and Iraq must consider a wide range of attack scenarios in addition to car bombings. Among these is the intentional poisoning of water supplies to obstruct military operations. To counter such attacks, the military is developing portable analyzers that can identify and quantify potential chemical agents in water supplies at microgram per liter concentrations within 10 minutes. To aid this effort we have been investigating the value of a surface-enhanced Raman spectroscopy based portable analyzer. In particular we have been developing silver-doped sol-gels to generate SER spectra of chemical agents and their hydrolysis products. Here we present SER spectra of methyl phosphonic acid and cyanide as a function of pH, an important factor affecting quantitation measurements, which to our knowledge has not been examined. In addition, dipicolinic acid, a chemical signature associated with anthrax-causing spores, is also presented.

  4. Continuous volatile fatty acid production from waste activated sludge hydrolyzed at pH 12.

    PubMed

    Yang, Xue; Wan, Chunli; Lee, Duu-Jong; Du, Maoan; Pan, Xiangliang; Wan, Fang

    2014-09-01

    This study adopted rapid alkaline treatment at pH 12 to hydrolyze 66% of total chemical oxygen demands. Then the hydrolyzed liquor was fermented in a continuous-flow stirred reactor to produce volatile fatty acids (VFAs) at 8-h hydraulic retention time and at 35 °C. The maximum VFA productivity reached 365 mg VFAs g(-1) volatile suspended solids in a 45-d operation, with most produced VFAs being acetate and propionate, principally produced by protein degradation. The Bacteroidia, ε-proteobacteria and the Clostridia were identified to be the classes correlating with the fermentation processes. The fermented liquor was applied to denitrifying phosphorus removal process as alternative carbon source after excess phosphorus and nitrogen being recycled via struvite precipitation. Fermented liquors from alkaline hydrolysis-acid fermentation on waste activated sludge are a potential renewable resource for applications that need organic carbons.

  5. Influence of amino acids, buffers, and ph on the γ-irradiation-induced degradation of alginates.

    PubMed

    Ulset, Ann-Sissel T; Mori, Hideki; Dalheim, Marianne Ø; Hara, Masayuki; Christensen, Bjørn E

    2014-12-08

    Alginate-based biomaterials and medical devices are commonly subjected to γ-irradiation as a means of sterilization, either in the dry state or the gel (hydrated) state. In this process the alginate chains degrade randomly in a dose-dependent manner, altering alginates' material properties. The addition of free radical scavenging amino acids such as histidine and phenylalanine protects the alginate significantly against degradation, as shown by monitoring changes in the molecular weight distributions using SEC-MALLS and determining the pseudo first order rate constants of degradation. Tris buffer (0.5 M), but not acetate, citrate, or phosphate buffers had a similar effect on the degradation rate. Changes in pH itself had only marginal effects on the rate of alginate degradation and on the protective effect of amino acids. Contrary to previous reports, the chemical composition (M/G profile) of the alginates, including homopolymeric mannuronan, was unaltered following irradiation up to 10 kGy.

  6. Comparing the acidities of aqueous, frozen, and freeze-dried phosphate buffers: Is there a "pH memory" effect?

    PubMed

    Vetráková, Ľubica; Vykoukal, Vít; Heger, Dominik

    2017-09-15

    The concept of "pH memory" has been established in the literature for the correlation between the pH of a pre-lyophilization solution and the ionization state of freeze-dried powder (lyophile). In this paper, the concept of "pH memory" is explored for the system of an aqueous solution, a frozen solution, and a lyophile. Sodium and potassium phosphate buffers in the pH range of 5-9 were frozen and lyophilized with sulfonephthalein indicators as acidity probes, and their Hammett acidity functions were compared to the initial pH of the aqueous solution. The results show that the acidities of the lyophiles are somewhat changed compared to the initial pHs, but the acidities in the frozen state differ more substantially. The Hammett acidity functions of the frozen buffers were found to be markedly dissimilar from the initial pH, especially in the sodium phosphate frozen at 233K, where an increase in the initial pH led to a decrease in the Hammett acidity function of the frozen state at a certain pH range. The large acidification observed after freezing the sodium phosphate buffer was not detected in the lyophiles after the sample had been dried; the phenomenon is explained considering the formed crystals analyzed by X-ray powder diffraction. The results suggest that monitoring the final acidity of a lyophile is not sufficient to predict all the acidity changes throughout the whole lyophilization process. The importance of well-controlled freezing and lyophilization conditions follows from the results of the research. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Different mechanisms of free fatty acid flip-flop and dissociation revealed by temperature and molecular species dependence of transport across lipid vesicles.

    PubMed

    Kampf, J Patrick; Cupp, David; Kleinfeld, Alan M

    2006-07-28

    The mechanism of free fatty acid (FFA) transport across membranes is a subject of intense investigation. We have demonstrated recently that flip-flop is the rate-limiting step for transport of oleic acid across phospholipid vesicles (Cupp, D., Kampf, J. P., and Kleinfeld, A. M. (2004) Biochemistry 43, 4473-4481). To better understand the nature of the flip-flop barrier, we measured the temperature dependence of a series of saturated and monounsaturated FFA. We determined the rate constants for flip-flop and dissociation for small (SUV), large (LUV), and giant (GUV) unilamellar vesicles composed of egg phosphatidylcholine. For all FFA and vesicle types, dissociation was faster than flip-flop, and for all FFA, flip-flop and dissociation were faster in SUV than in LUV or GUV. Rate constants for both flip-flop and dissociation decreased exponentially with increasing FFA size. However, only the flip-flop rate constants increased significantly with temperature; the barrier to flip-flop was virtually entirely due to an enthalpic activation free energy. The barrier to dissociation was primarily entropic. Analysis in terms of a simple free volume (V(f)) model revealed V(f) values for flip-flop that ranged between approximately 12 and 15 Angstroms(3), with larger values for SUV than for LUV or GUV. V(f) values increased with temperature, and this temperature dependence generated the enthalpic barrier to flip-flop. The barrier for dissociation and its size dependence primarily reflect the aqueous solubility of FFA. These are the first results to distinguish the energetics of flipflop and dissociation. This should lead to a better understanding of the mechanisms governing FFA transport across biological membranes.

  8. A pH dependent Raman and surface enhanced Raman spectroscopic studies of citrazinic acid aided by theoretical calculations.

    PubMed

    Sarkar, Sougata; Chowdhury, Joydeep; Dutta, Soumen; Pal, Tarasankar

    2016-12-05

    A pH dependent normal Raman scattering (NRS) and surface enhanced Raman scattering (SERS) spectral patterns of citrazinic acid (CZA), a biologically important molecule, have been investigated. The acid, with different pKa values (~4 and ~11) for the two different functional groups (-COOH and -OH groups), shows interesting range of color changes (yellow at pH~14 and brown at pH~2) with the variation in solution pH. Thus, depending upon the pH of the medium, CZA molecule can exist in various protonated and/or deprotonated forms. Here we have prescribed the existence different possible forms of CZA at different pH (Forms "C", "H" and "Dprot" at pH~14 and Forms "A", "D", and "P" at pH~2 respectively). The NRS spectra of these solutions and their respective SERS spectra over gold nanoparticles were recorded. The spectra clearly differ in their spectral profiles. For example the SERS spectra recorded with the CZA solution at pH~2 shows blue shift for different bands compared to its NRS window e.g. 406 to 450cm(-1), 616 to 632cm(-1), 1332 to 1343cm(-1) etc. Again, the most enhanced peak at ~1548cm(-1) in NRS while in the SERS window this appears at ~1580cm(-1). Similar observation was also made for CZA at pH~14. For example, the 423cm(-1) band in the NRS profile experience a blue shift and appears at ~447cm(-1) in the SERS spectrum as well as other bands at ~850, ~1067 and ~1214cm(-1) in the SERS window are markedly enhanced. It is also worth noting that the SERS spectra at the different pH also differ from each other. These spectral differences indicate the existence of various adsorptive forms of the CZA molecule depending upon the pH of the solution. Therefore based on the experimental findings we propose different possible molecular forms of CZA at different pH (acidic and alkaline) conditions. For example forms 'A', 'D' and 'P' existing in acidic pH (pH~2) and three other deprotonated forms 'C', 'H' and 'Dprot' in alkaline pH (pH~14). The DFT calculations for these

  9. Acidic pH resistance of grafted chitosan on dental implant.

    PubMed

    Campos, Doris M; Toury, Bérengère; D'Almeida, Mélanie; Attik, Ghania N; Ferrand, Alice; Renoud, Pauline; Grosgogeat, Brigitte

    2015-05-01

    Over the last decade, access to dental care has increasingly become a service requested by the population, especially in the case of dental implants. However, the major cause of implant failure is an inflammatory disease: peri-implantitis. Currently, the adhesion strength of antibacterial coatings at implant surfaces remains a problem to solve. In order to propose a functionalized implant with a resistant antibacterial coating, a novel method of chitosan immobilization at implant surface has been investigated. Functionalization of the pre-active titanium (Ti) surface was performed using triethoxysilylpropyl succinic anhydride (TESPSA) as a coupling agent which forms a stable double peptide bond with chitosan. The chitosan presence and the chemical resistibility of the coating under acid pH solutions (pH 5 and pH 3) were confirmed by FTIR-ATR and XPS analyses. Furthermore, peel test results showed high adhesive resistance of the TESPSA/chitosan coating at the substrate. Cytocompatibility was evaluated by cell morphology with confocal imaging. Images showed healthy morphology of human gingival fibroblasts (HGF-1). Finally, the reported method for chitosan immobilization on Ti surface via peptide bindings allows for the improvement of its adhesive capacities and resistibility while maintaining its cytocompatibility. Surface functionalization using the TESPSA/chitosan coupling method is noncytotoxic and stable even in drastic environments as found in oral cavity, thus making it a valuable candidate for clinical implantology applications.

  10. Effect of pH, substrate and free nitrous acid concentrations on ammonium oxidation rate.

    PubMed

    Jiménez, E; Giménez, J B; Seco, A; Ferrer, J; Serralta, J

    2012-11-01

    Respirometric techniques have been used to determine the effect of pH, free nitrous acid (FNA) and substrate concentration on the activity of the ammonium oxidizing bacteria (AOB) present in an activated sludge reactor. With this aim, bacterial activity has been measured at different pH values (ranging from 6.2 to 9.7), total ammonium nitrogen concentrations (ranging from 0.1 to 10 mg TAN L(-1)) and total nitrite concentrations (ranging from 3 to 43 mg NO(2)-NL(-1)). According to the results obtained, the most appropriate kinetic expression for the growth of AOB in activated sludge reactors has been established. Substrate half saturation constant and FNA and pH inhibition constants have been obtained by adjusting model predictions to experimental results. Different kinetic parameter values and different Monod terms should be used to model the growth of AOB in activated sludge processes and SHARON reactors due to the different AOB species that predominate in both systems.

  11. Association of the pr Peptides with Dengue Virus at Acidic pH Blocks Membrane Fusion

    SciTech Connect

    Yu, I.-M.; Holdaway, H.A.; Chipman, P.R.; Kuhn, R.J.; Rossmann, M.G.; Chen, J.; Purdue

    2010-07-27

    Flavivirus assembles into an inert particle that requires proteolytic activation by furin to enable transmission to other hosts. We previously showed that immature virus undergoes a conformational change at low pH that renders it accessible to furin (I. M. Yu, W. Zhang, H. A. Holdaway, L. Li, V. A. Kostyuchenko, P. R. Chipman, R. J. Kuhn, M. G. Rossmann, and J. Chen, Science 319:1834-1837, 2008). Here we show, using cryoelectron microscopy, that the structure of immature dengue virus at pH 6.0 is essentially the same before and after the cleavage of prM. The structure shows that after cleavage, the proteolytic product pr remains associated with the virion at acidic pH, and that furin cleavage by itself does not induce any major conformational changes. We also show by liposome cofloatation experiments that pr retention prevents membrane insertion, suggesting that pr is present on the virion in the trans-Golgi network to protect the progeny virus from fusion within the host cell.

  12. Robust Extracellular pH Modulation by Candida albicans during Growth in Carboxylic Acids

    PubMed Central

    Danhof, Heather A.; Vylkova, Slavena; Vesely, Elisa M.; Ford, Amy E.; Gonzalez-Garay, Manuel

    2016-01-01

    ABSTRACT The opportunistic fungal pathogen Candida albicans thrives within diverse niches in the mammalian host. Among the adaptations that underlie this fitness is an ability to utilize a wide array of nutrients, especially sources of carbon that are disfavored by many other fungi; this contributes to its ability to survive interactions with the phagocytes that serve as key barriers against disseminated infections. We have reported that C. albicans generates ammonia as a byproduct of amino acid catabolism to neutralize the acidic phagolysosome and promote hyphal morphogenesis in a manner dependent on the Stp2 transcription factor. Here, we report that this species rapidly neutralizes acidic environments when utilizing carboxylic acids like pyruvate, α-ketoglutarate (αKG), or lactate as the primary carbon source. Unlike in cells growing in amino acid-rich medium, this does not result in ammonia release, does not induce hyphal differentiation, and is genetically distinct. While transcript profiling revealed significant similarities in gene expression in cells grown on either carboxylic or amino acids, genetic screens for mutants that fail to neutralize αKG medium identified a nonoverlapping set of genes, including CWT1, encoding a transcription factor responsive to cell wall and nitrosative stresses. Strains lacking CWT1 exhibit retarded αKG-mediated neutralization in vitro, exist in a more acidic phagolysosome, and are more susceptible to macrophage killing, while double cwt1Δ stp2Δ mutants are more impaired than either single mutant. Together, our observations indicate that C. albicans has evolved multiple ways to modulate the pH of host-relevant environments to promote its fitness as a pathogen. PMID:27935835

  13. Purification of coated vesicles by agarose gel electrophoresis

    PubMed Central

    1981-01-01

    We have applied agarose gel electrophoresis as a novel step in the purification of clathrin-coated vesicles. Preparations of coated vesicles obtained by sedimentation velocity and isopycnic centrifugation are resolved into two distinct fractions upon electrophoresis. The slower migrating fraction contains smooth vesicles, whereas the faster contains only coated vesicles and empty clathrin coats. The faster mobility of the coated vesicles is primarily caused by the acidic nature of clathrin. Coated vesicles from three different cell types have different mobilities. In each case, however, all of the major polypeptides previously attributed to coated vesicles comigrate with the now homogeneous particles, even though a powerful ATPase activity is completely removed. PMID:6265464

  14. The effect of pH on the toxicity of fatty acids and fatty acid amides to rainbow trout gill cells.

    PubMed

    Bertin, Matthew J; Voronca, Delia C; Chapman, Robert W; Moeller, Peter D R

    2014-01-01

    Harmful algal blooms (HABs) expose aquatic organisms to multiple physical and chemical stressors during an acute time period. Algal toxins themselves may be altered by water chemistry parameters affecting their bioavailability and resultant toxicity. The purpose of this study was to determine the effects of two abiotic parameters (pH, inorganic metal salts) on the toxicity of fatty acid amides and fatty acids, two classes of lipids produced by harmful algae, including the golden alga, Prymnesium parvum, that are toxic to aquatic organisms. Rainbow trout gill cells were used as a model of the fish gill and exposed to single compounds and mixtures of compounds along with variations in pH level and concentration of inorganic metal salts. We employed artificial neural networks (ANNs) and standard ANOVA statistical analysis to examine and predict the effects of these abiotic parameters on the toxicity of fatty acid amides and fatty acids. Our results demonstrate that increasing pH levels increases the toxicity of fatty acid amides and inhibits the toxicity of fatty acids. This phenomenon is reversed at lower pH levels. Exposing gill cells to complex mixtures of chemical factors resulted in dramatic increases in toxicity compared to tests of single compounds for both the fatty acid amides and fatty acids. These findings highlight the potential of physicochemical factors to affect the toxicity of chemicals released during algal blooms and demonstrate drastic differences in the effect of pH on fatty acid amides and fatty acids.

  15. Changes in composition and enamel demineralization inhibition activities of gallic acid at different pH values.

    PubMed

    Zhang, Jingyang; Huang, Xuelian; Huang, Shengbin; Deng, Meng; Xie, Xincheng; Liu, Mingdong; Liu, Hongling; Zhou, Xuedong; Li, Jiyao; Ten Cate, Jacob Martien

    2015-01-01

    Gallic acid (GA) has been shown to inhibit demineralization and enhance remineralization of enamel; however, GA solution is highly acidic. This study was to investigate the stability of GA solutions at various pH and to examine the resultant effects on enamel demineralization. The stability of GA in H2O or in phosphate buffer at pH 5.5, pH 7.0 and pH 10.0 was evaluated qualitatively by ultraviolet absorption spectra and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Then, bovine enamel blocks were subjected to a pH-cycling regime of 12 cycles. Each cycle included 5 min applications with one of the following treatments: 1 g/L NaF (positive control), 4 g/L GA in H2O or buffered at pH 5.5, pH 7.0 and pH 10.0 and buffers without GA at the same pH (negative control), followed by a 60 min application with pH 5.0 acidic buffers and a 5 min application with neutral buffers. The acidic buffers were analysed for dissolved calcium. GA was stable in pure water and acidic condition, but was unstable in neutral and alkaline conditions, in which ultraviolet spectra changed and HPLC-DAD analysis revealed that most of the GA was degraded. All the GA groups significantly inhibited demineralization (p < 0.05) and there was no significant difference of the inhibition efficacy among different GA groups (p > 0.05). GA could inhibit enamel demineralization and the inhibition effect is not influenced by pH. GA could be a useful source as an anti-cariogenic agent for broad practical application.

  16. Estimated net acid excretion inversely correlates with urine pH in vegans, lacto-ovo vegetarians, and omnivores.

    PubMed

    Ausman, Lynne M; Oliver, Lauren M; Goldin, Barry R; Woods, Margo N; Gorbach, Sherwood L; Dwyer, Johanna T

    2008-09-01

    Diet affects urine pH and acid-base balance. Both excess acid/alkaline ash (EAA) and estimated net acid excretion (NAE) calculations have been used to estimate the effects of diet on urine pH. This study's goal was to determine if free-living vegans, lacto-ovo vegetarians, and omnivores have increasingly acidic urine, and to assess the ability of EAA and estimated NAE calculations to predict urine pH. This study used a cross-sectional design. This study assessed urine samples of 10 vegan, 16 lacto-ovo vegetarian, and 16 healthy omnivorous women in the Boston metropolitan area. Six 3-day food records from each dietary group were analyzed for EAA content and estimated NAE, and correlations with measured urine pH were calculated. The mean (+/- SD) urine pH was 6.15 +/- 0.40 for vegans, 5.90 +/- 0.36 for lacto-ovo vegetarians, and 5.74 +/- 0.21 for omnivores (analysis of variance, P = .013). Calculated EAA values were not significantly different among the three groups, whereas mean estimated NAE values were significantly different: 17.3 +/- 14.5 mEq/day for vegans, 31.3 +/- 8.5 mEq/day for lacto-ovo vegetarians, and 42.6 +/- 13.2 mEq/day for omnivores (analysis of variance, P = .01). The average deattenuated correlation between urine pH and EAA was 0.333; this value was -0.768 for estimated NAE and urine pH, with a regression equation of pH = 6.33 - 0.014 NAE (P = .02, r = -0.54). Habitual diet and estimated NAE calculations indicate the probable ranking of urine pH by dietary groups, and may be used to determine the likely acid-base status of an individual; EAA calculations were not predictive of urine pH.

  17. Optogenetic Acidification of Synaptic Vesicles and Lysosomes

    PubMed Central

    Grauel, M. Katharina; Wozny, Christian; Bentz, Claudia; Blessing, Anja; Rosenmund, Tanja; Jentsch, Thomas J.; Schmitz, Dietmar; Hegemann, Peter; Rosenmund, Christian

    2016-01-01

    Acidification is required for the function of many intracellular organelles, but methods to acutely manipulate their intraluminal pH have not been available. Here we present a targeting strategy to selectively express the light-driven proton pump Arch3 on synaptic vesicles. Our new tool, pHoenix, can functionally replace endogenous proton pumps, enabling optogenetic control of vesicular acidification and neurotransmitter accumulation. Under physiological conditions, glutamatergic vesicles are nearly full, as additional vesicle acidification with pHoenix only slightly increased the quantal size. By contrast, we found that incompletely filled vesicles exhibited a lower release probability than full vesicles, suggesting preferential exocytosis of vesicles with high transmitter content. Our subcellular targeting approach can be transferred to other organelles, as demonstrated for a pHoenix variant that allows light-activated acidification of lysosomes. PMID:26551543

  18. Thermodynamics of the interaction of globular proteins with powdered stearic acid in acid pH.

    PubMed

    Mitra, Atanu; Chattoraj, D K; Chakraborty, P

    2006-06-01

    Adsorption isotherms of different globular proteins and gelatin on strearic acid particles have been studied as a function of biopolymer concentration, ionic strength of the medium, and temperature. The effect of neutral salts including CaCl2, Na3PO4, and urea on the adsorption isotherms has been also investigated. It is observed that the extent of adsorption (Gamma2(1)) increases in two steps with the increase of biopolymer concentration (C2) in the bulk. Gamma2(1) increases with an increase of C2 until a steady maximum value Gamma2(m) is reached at a critical concentration C2(m). After initial saturation, Gamma2(1) again increases from Gamma2(m) without reaching any limiting value due to the surface aggregation of the protein. The values of the standard free energy change for adsorption have been calculated on the basis of the Gibbs equation. The standard entropy and enthalpy changes are also calculated.

  19. TRP-ML1 regulates lysosomal pH and acidic lysosomal lipid hydrolytic activity.

    PubMed

    Soyombo, Abigail A; Tjon-Kon-Sang, Sandra; Rbaibi, Youssef; Bashllari, Enkelejda; Bisceglia, Jill; Muallem, Shmuel; Kiselyov, Kirill

    2006-03-17

    Mucolipidosis type IV (MLIV) is caused by mutations in the ion channel mucolipin 1 (TRP-ML1). MLIV is typified by accumulation of lipids and membranous materials in intracellular organelles, which was hypothesized to be caused by the altered membrane fusion and fission events. How mutations in TRP-ML1 lead to aberrant lipolysis is not known. Here we present evidence that MLIV is a metabolic disorder that is not associated with aberrant membrane fusion/fission events. Thus, measurement of lysosomal pH revealed that the lysosomes in TRP-ML1(-/-) cells obtained from the patients with MLIV are over-acidified. TRP-ML1 can function as a H(+) channel, and the increased lysosomal acidification in TRP-ML1(-/-) cells is likely caused by the loss of TRP-ML1-mediated H(+) leak. Measurement of lipase activity using several substrates revealed a marked reduction in lipid hydrolysis in TRP-ML1(-/-) cells, which was rescued by the expression of TRP-ML1. Cell fractionation indicated specific loss of acidic lipase activity in TRP-ML1(-/-) cells. Furthermore, dissipation of the acidic lysosomal pH of TRP-ML1(-/-) cells by nigericin or chloroquine reversed the lysosomal storage disease phenotype. These findings provide a new mechanism to account for the pathogenesis of MLIV.

  20. Do pH and flavonoids influence hypochlorous acid-induced catalase inhibition and heme modification?

    PubMed

    Krych-Madej, Justyna; Gebicka, Lidia

    2015-09-01

    Hypochlorous acid (HOCl), highly reactive oxidizing and chlorinating species, is formed in the immune response to invading pathogens by the reaction of hydrogen peroxide with chloride catalyzed by the enzyme myeloperoxidase. Catalase, an important antioxidant enzyme, catalyzing decomposition of hydrogen peroxide to water and molecular oxygen, hampers in vitro HOCl formation, but is also one of the main targets for HOCl. In this work we have investigated HOCl-induced catalase inhibition at different pH, and the influence of flavonoids (catechin, epigallocatechin gallate and quercetin) on this process. It has been shown that HOCl-induced catalase inhibition is independent on pH in the range 6.0-7.4. Preincubation of catalase with epigallocatechin gallate and quercetin before HOCl treatment enhances the degree of catalase inhibition, whereas catechin does not affect this process. Our rapid kinetic measurements of absorption changes around the heme group have revealed that heme modification by HOCl is mainly due to secondary, intramolecular processes. The presence of flavonoids, which reduce active catalase intermediate, Compound I to inactive Compound II have not influenced the kinetics of HOCl-induced heme modification. Possible mechanisms of the reaction of hypochlorous acid with catalase are proposed and the biological consequences are discussed. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Poly methacrylic acid modified CDHA nanocomposites as potential pH responsive drug delivery vehicles.

    PubMed

    Victor, Sunita Prem; Sharma, Chandra P

    2013-08-01

    The objective of this study was to prepare pH sensitive polymethacrylic acid-calcium deficient hydroxyapatite (CDHA) nanocomposites. The CDHA nanoparticles were prepared by coprecipitation method. The modification of CDHA by methacrylic acid (MA) was achieved by AIBN initiated free radical polymerization with sodium bisulphite as catalyst followed by emulsion technique. These nanocomposites with a half life of 8h consisted of high aspect ratio, needle like particles and exhibited an increase in swelling behaviour with pH. The in vivo potential of the nanocomposites was evaluated in vitro by the results of cell aggregation, protein adsorption, MTT assay and haemolytic activity. The invitro loading and release studies using albumin as a model drug indicate that the nanocomposites gave better loading when compared to the CDHA nanoparticles and altered the drug release rates. The nanocomposites also exhibited good uptake on C6 glioma cells as studied by fluorescence microscopy. The results obtained suggest that these nanocomposites have great potential for oral controlled protein delivery and can be extended further for intracellular drug delivery applications.

  2. Acidic pH retards the fibrillization of human islet amyloid polypeptide due to electrostatic repulsion of histidines

    NASA Astrophysics Data System (ADS)

    Li, Yang; Xu, Weixin; Mu, Yuguang; Zhang, John Z. H.

    2013-08-01

    The human Islet Amyloid Polypeptide (hIAPP) is the major constituent of amyloid deposits in pancreatic islets of type-II diabetes. IAPP is secreted together with insulin from the acidic secretory granules at a low pH of approximately 5.5 to the extracellular environment at a neutral pH. The increased accumulation of extracellular hIAPP in diabetes indicates that changes in pH may promote amyloid formation. To gain insights and underlying mechanisms of the pH effect on hIAPP fibrillogenesis, all-atom molecular dynamics simulations in explicit solvent model were performed to study the structural properties of five hIAPP protofibrillar oligomers, under acidic and neutral pH, respectively. In consistent with experimental findings, simulation results show that acidic pH is not conducive to the structural stability of these oligomers. This provides a direct evidence for a recent experiment [L. Khemtemourian, E. Domenech, J. P. F. Doux, M. C. Koorengevel, and J. A. Killian, J. Am. Chem. Soc. 133, 15598 (2011)], 10.1021/ja205007j, which suggests that acidic pH inhibits the fibril formation of hIAPP. In addition, a complementary coarse-grained simulation shows the repulsive electrostatic interactions among charged His18 residues slow down the dimerization process of hIAPP by twofold. Besides, our all-atom simulations reveal acidic pH mainly affects the local structure around residue His18 by destroying the surrounding hydrogen-bonding network, due to the repulsive interactions between protonated interchain His18 residues at acidic pH. It is also disclosed that the local interactions nearby His18 operating between adjacent β-strands trigger the structural transition, which gives hints to the experimental findings that the rate of hIAPP fibril formation and the morphologies of the fibrillar structures are strongly pH-dependent.

  3. Membrane vesicles: A simplified system for studying auxin transport

    SciTech Connect

    Goldsmith, M.H.M.

    1989-01-01

    Indoleacetic acid (IAA), the auxin responsible for regulation of growth, is transported polarly in plants. Several different models have been suggested to account for IAA transport by cells and its accumulation by membrane vesicles. One model sees diffusion of IAA driven by a pH gradient. The anion of a lipophilic weak acid like IAA or butyrate accumulates in an alkaline compartment in accord with the size of the pH gradient The accumulation of IAA may be diminished by the permeability of its lipophilic anion. This anion leak may be blocked by NPA. With anion efflux blocked, a gradient of two pH units would support an IAA accumulation of less than 50-fold at equilibrium (2) Another model sees diffusion of IAA in parallel with a saturable symport (IAA[sup [minus

  4. Influence of glutamic acid residues and pH on the properties of transmembrane helices.

    PubMed

    Rajagopalan, Venkatesan; Greathouse, Denise V; Koeppe, Roger E

    2017-03-01

    Negatively charged side chains are important for the function of particular ion channels and certain other membrane proteins. To investigate the influence of single glutamic acid side chains on helices that span lipid-bilayer membranes, we have employed GWALP23 (acetyl-GGALW(5)LALALALALALALW(19)LAGA-amide) as a favorable host peptide framework. We substituted individual Leu residues with Glu residues (L12E or L14E or L16E) and incorporated specific (2)H-labeled alanine residues within the core helical region or near the ends of the sequence. Solid-state (2)H NMR spectra reveal little change for the core labels in GWALP23-E12, -E14 and -E16 over a pH range of 4 to 12.5, with the spectra being broader for samples in DOPC compared to DLPC bilayers. The spectra for samples with deuterium labels near the helix ends on alanines 3 and 21 show modest pH-dependent changes in the extent of unwinding of the helix terminals in DLPC and DOPC bilayers. The combined results indicate minor overall responses of these transmembrane helices to changes in pH, with the most buried residue E12 showing no pH dependence. While the Glu residues E14 and E16 may have high pKa values in the lipid bilayer environment, it is also possible that a paucity of helix response is masking the pKa values. Interestingly, when E16 is present, spectral changes at high pH report significant local unwinding of the core helix. Our results are consistent with the expectation that buried carboxyl groups aggressively hold their protons and/or waters of hydration.

  5. pH Titratable Superparamagnetic Iron Oxide for Improved Nanoparticle Accumulation in Acidic Tumor Microenvironments

    PubMed Central

    Crayton, Samuel H.; Tsourkas, Andrew

    2011-01-01

    A wide variety of nanoparticle platforms are being developed for the diagnosis and treatment of malignancy. While many of these are passively targeted or rely on receptor-ligand interactions, metabolically directed nanoparticles provide a complementary approach. It is known that both primary and secondary events in tumorigensis alter the metabolic profile of developing and metastatic cancers. One highly conserved metabolic phenotype is a state of up-regulated glycolysis and reduced use of oxidative phosphorylation, even when oxygen tension is not limiting. This metabolic shift, termed the Warburg effect, creates a “hostile” tumor microenvironment with increased levels of lactic acid and low extracellular pH. In order to exploit this phenomenon and improve the delivery of nanoparticle platforms to a wide variety of tumors, a pH-responsive iron oxide nanoparticle was designed. Specifically, glycol chitosan (GC), a water-soluble polymer with pH titratable charge, was conjugated to the surface of superparamagnetic iron oxide nanoparticles (SPIO) to generate a T2*-weighted MR contrast agent that responds to alterations in its surrounding pH. Compared to control nanoparticles that lack pH sensitivity, these GC-SPIO nanoparticles demonstrated potent pH-dependent cellular association and MR contrast in vitro. In murine tumor models GC-SPIO also generated robust T2*-weighted contrast, which correlated with increased delivery of the agent to the tumor site, measured quantitatively by inductively coupled plasma mass spectrometry. Importantly, the increased delivery of GC-SPIO nanoparticles cannot be solely attributed to the commonly observed enhanced permeability and retention effect, since these nanoparticles have similar physical properties and blood circulation times as control agents. PMID:22035454

  6. Microbial sulfate reduction and metal attenuation in pH 4 acid mine water

    USGS Publications Warehouse

    Church, C.D.; Wilkin, R.T.; Alpers, C.N.; Rye, R.O.; Blaine, R.B.

    2007-01-01

    Sediments recovered from the flooded mine workings of the Penn Mine, a Cu-Zn mine abandoned since the early 1960s, were cultured for anaerobic bacteria over a range of pH (4.0 to 7.5). The molecular biology of sediments and cultures was studied to determine whether sulfate-reducing bacteria (SRB) were active in moderately acidic conditions present in the underground mine workings. Here we document multiple, independent analyses and show evidence that sulfate reduction and associated metal attenuation are occurring in the pH-4 mine environment. Water-chemistry analyses of the mine water reveal: (1) preferential complexation and precipitation by H2S of Cu and Cd, relative to Zn; (2) stable isotope ratios of 34S/32S and 18O/16O in dissolved SO4 that are 2-3 ??? heavier in the mine water, relative to those in surface waters; (3) reduction/oxidation conditions and dissolved gas concentrations consistent with conditions to support anaerobic processes such as sulfate reduction. Scanning electron microscope (SEM) analyses of sediment show 1.5-micrometer, spherical ZnS precipitates. Phospholipid fatty acid (PLFA) and denaturing gradient gel electrophoresis (DGGE) analyses of Penn Mine sediment show a high biomass level with a moderately diverse community structure composed primarily of iron- and sulfate-reducing bacteria. Cultures of sediment from the mine produced dissolved sulfide at pH values near 7 and near 4, forming precipitates of either iron sulfide or elemental sulfur. DGGE coupled with sequence and phylogenetic analysis of 16S rDNA gene segments showed populations of Desulfosporosinus and Desulfitobacterium in Penn Mine sediment and laboratory cultures. ?? 2007 Church et al; licensee BioMed Central Ltd.

  7. Quantification of humic acids in surface water: effects of divalent cations, pH, and filtration.

    PubMed

    Rodrigues, Alexandrina; Brito, António; Janknecht, Peter; Proença, Maria Fernanda; Nogueira, Regina

    2009-02-01

    Humic acids (HAs) content of raw water is an important analytical parameter in water treatment facilities because HAs in the presence of chlorine may lead to the formation of dangerous by-products (e.g., trihalomethanes). The concentration of HAs in water is not directly accessible by common analytical methods due to their heterogeneous chemical structure. The aim of this study was to compare two methods to assess humic acids (HAs) in surface water namely absorbance of ultraviolet light at 254 nm (UV(254)) and total organic carbon (TOC), as well as to evaluate the effects of calcium and magnesium concentrations, pH and sample filtration on the methods' results. An aqueous solution of a commercial HA with 10 mg L(-1) was used in the present work. Quantification of the HA was carried out by both UV(254) and TOC (combustion-infrared method) measurements. UV(254) results were converted to TOC using a calibration curve. The effects of calcium (0-136.3 mg L(-1)) and magnesium (0-34.5 mg L(-1)) concentrations, pH (4.0, 7.0 and 9.0) and sample filtration on UV(254) and TOC measurements of the HA suspension were evaluated. More accurate TOC values of HA suspensions were obtained by the combustion-infrared method than by the UV(254) absorbance method. The higher differences of TOC values between unfiltered and filtered samples were detected in the presence of calcium at pH 9.0 using the spectrophotometric method.

  8. [Effects of simulated acid rain on respiration rate of cropland system with different soil pH].

    PubMed

    Zhu, Xue-zhu; Zhang, Gao-chuan; Li, Hui

    2009-10-15

    To evaluate the effects of acid rain on the respiration rate of cropland system, an outdoor pot experiment was conducted with paddy soils of pH 5.48 (S1), pH 6.70 (S1) and pH 8.18 (S3) during the 2005-2007 wheat-growing seasons. The cropland system was exposed to acid rain by spraying the wheat foliage and irrigating the soil with simulated rainwater of T1 (pH 6.0), T2 (pH 6.0, ionic concentration was twice as rainwater T1), and T3 (pH 4.4, ionic concentration was twice as rainwater T1), respectively. The static opaque chamber-gas chromatograph method was used to measure CO2 fluxes from cropland system. The results showed that acid rain affected the respiration rate of cropland system through crop plant, and the cropland system could adapt to acid rain. Acid rainwater significantly increased the average respiration rate in alkaline soil (S3) cropland system, while it had no significant effects on the average respiration rate in neutral soil (S2) and acidic soil (S1) cropland systems. During 2005-2006, after the alkaline soil cropland system was treated with rainwater T3, the average respiration rate was 23.6% and 27.6% higher than that of alkaline soil cropland system treated with rainwater T1 and T2, respectively. During March to April, the respiration rate was enhanced with the increase of rainwater ionic concentration, while it was dropped with the decrease of rainwater pH value in acidic soil cropland system. It was demonstrated that soil pH and crop plant played important roles on the respiration rate of cropland system.

  9. Predicting Thermodynamic Behaviors of Non-Protein Amino Acids as a Function of Temperature and pH.

    PubMed

    Kitadai, Norio

    2016-03-01

    Why does life use α-amino acids exclusively as building blocks of proteins? To address that fundamental question from an energetic perspective, this study estimated the standard molal thermodynamic data for three non-α-amino acids (β-alanine, γ-aminobutyric acid, and ε-aminocaproic acid) and α-amino-n-butyric acid in their zwitterionic, negative, and positive ionization states based on the corresponding experimental measurements reported in the literature. Temperature dependences of their heat capacities were described based on the revised Helgeson-Kirkham-Flowers (HKF) equations of state. The obtained dataset was then used to calculate the standard molal Gibbs energies (∆G (o)) of the non-α-amino acids as a function of temperature and pH. Comparison of their ∆G (o) values with those of α-amino acids having the same molecular formula showed that the non-α-amino acids have similar ∆G (o) values to the corresponding α-amino acids in physiologically relevant conditions (neutral pH, <100 °C). In acidic and alkaline pH, the non-α-amino acids are thermodynamically more stable than the corresponding α-ones over a broad temperature range. These results suggest that the energetic cost of synthesis is not an important selection pressure to incorporate α-amino acids into biological systems.

  10. Predicting Thermodynamic Behaviors of Non-Protein Amino Acids as a Function of Temperature and pH

    NASA Astrophysics Data System (ADS)

    Kitadai, Norio

    2016-03-01

    Why does life use α-amino acids exclusively as building blocks of proteins? To address that fundamental question from an energetic perspective, this study estimated the standard molal thermodynamic data for three non-α-amino acids (β-alanine, γ-aminobutyric acid, and ɛ-aminocaproic acid) and α-amino- n-butyric acid in their zwitterionic, negative, and positive ionization states based on the corresponding experimental measurements reported in the literature. Temperature dependences of their heat capacities were described based on the revised Helgeson-Kirkham-Flowers (HKF) equations of state. The obtained dataset was then used to calculate the standard molal Gibbs energies ( ∆G o) of the non-α-amino acids as a function of temperature and pH. Comparison of their ∆G o values with those of α-amino acids having the same molecular formula showed that the non-α-amino acids have similar ∆G o values to the corresponding α-amino acids in physiologically relevant conditions (neutral pH, <100 °C). In acidic and alkaline pH, the non-α-amino acids are thermodynamically more stable than the corresponding α-ones over a broad temperature range. These results suggest that the energetic cost of synthesis is not an important selection pressure to incorporate α-amino acids into biological systems.

  11. Ru-η(6) -arene cations [{(Ph2 PC6 H4 )2 B(η(6) -Ph)}RuX](+) (X=Cl, H) as Lewis acids.

    PubMed

    Boone, Michael P; Stephan, Douglas W

    2014-03-17

    The reactivity of [{(Ph2 PC6 H4 )2 B(η(6) -Ph)}RuCl][B(C6 F5 )4 ] (1) as a Lewis acid was investigated. Treatment of 1 with mono and multidentate phosphorus Lewis bases afforded the Lewis acid-base adducts with the ortho-carbon atom of the coordinated arene ring. Similar reactivity was observed upon treatment with N-heterocyclic carbenes; however, adduct formation occurred at both ortho- and para-carbon atoms of the bound arene with the para-position being favoured by increased steric demands. Interestingly treatment with isocyanides resulted in adduct formation with the B-centre of the ligand framework. The hydride-cation [{(Ph2 PC6 H4 )2 B(η(6) -Ph)}RuH] [B(C6 F5 )4 ] was prepared via reaction of 1 with silane. This species in the presence of a bulky phosphine behaves as a frustrated Lewis pair (FLP) to activate H2 between the phosphorus centre and the ortho-carbon atom of the η(6) -arene ring. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Enzymatic characterization of peptidic materials isolated from aqueous solutions of ammonium cyanide (pH 9) and hydrocyanic acid (pH 6) exposed to ionizing radiation.

    PubMed

    Niketic, V; Draganić, Z; Nesković, S; Draganić, I

    1982-01-01

    The enzymatic digestion of some radiolytically produced peptidic materials was examined. The substrates were compounds isolated from 0.1 molar solutions of NH4CN (pH 9) and HCN (pH 6), after their exposure to gamma rays from a 60Co source (15-20 Mrad doses). Commercial proteolytic enzymes pronase and aminopeptidase M were used. The examined materials were of composite nature and proteolytic action was systematically observed after their subsequent purification. In some fractions the effect was found to be positive with up to 30% of peptide bonds cleaved with respect to the amino acid content. These findings support our previous conclusions on the free radical induced formation of peptidic backbones without the intervention of amino acids. Some side effects were also noted which might be of interest in observations on enzymatic cleavage of other composite peptidic materials of abiotic origin.

  13. Faecal pH, bile acid and sterol concentrations in premenopausal Indian and white vegetarians compared with white omnivores.

    PubMed

    Reddy, S; Sanders, T A; Owen, R W; Thompson, M H

    1998-06-01

    Faecal bulk, pH, water content, the concentrations of neutral sterols and bile acids and dietary intakes were measured in twenty-two Indian vegetarian, twenty-two white omnivorous and eighteen white vegetarian premenopausal women. Faecal bulk and water content were greater and pH lower in the Indian vegetarians. Total faecal animal sterol and coprostanol concentrations expressed on a dry-weight basis were lower in the vegetarians compared with the omnivores. The faecal sterol concentrations were correlated with dietary cholesterol intake. Primary bile acids were detected in six Indian vegetarians, two white vegetarians and two white omnivores; secondary bile acids were detected in all the white omnivores and vegetarian subjects but not in two of the Indian vegetarians. Total faecal free bile acid and conjugated bile acid concentrations were lower in the white vegetarians compared with the omnivores. Faecal lithocholic acid concentrations were lower in both Indian and white vegetarians. The lithocholic: deoxycholic acid ratio and coprostanol: total animal sterols ratio were significantly lower in the Indian vegetarians compared with the omnivores. Both ratios were positively correlated with faecal pH. Stepwise multiple regression analyses were undertaken in order to identify which nutrients influenced faecal pH, lithocholic and deoxycholic acid concentrations. The intakes of starch and dietary fibre were negatively associated with faecal concentrations of lithocholic and deoxycholic acid. Starch intake alone was negatively associated with faecal pH. The results of this study confirm that diets high in dietary fibre decrease faecal bile acid concentrations and suggest that the complex carbohydrates present in Indian vegetarian diets influence faecal pH and inhibit the degradation of faecal steroids.

  14. Involvement of vH+-ATPase in synaptic vesicle swelling

    PubMed Central

    Shin, Leah; Basi, Nirukti; Jeremic, Aleksandar; Lee, Jin-Sook; Cho, Won Jin; Chen, ZhiHui; Abu-Hamdah, Rania; Oupicky, David; Jena, Bhanu P

    2010-01-01

    Secretory vesicle swelling is central to cell secretion, however the underlying mechanism of vesicle swelling, particularly synaptic vesicles, is not completely understood. The Gαi3-PLA2-mediated involvement of water channel AQP-1 in the regulation of secretory vesicle swelling in exocrine pancreas, and the Gαo-mediated AQP-6 involvement in synaptic vesicle swelling in neurons, has previously been reported. Furthermore, the role of vH+-ATPase in neurotransmitter transport into synaptic vesicles, has also been shown. Using nanometer scale precision measurements of isolated synaptic vesicles, the present study reports for the first time, the involvement of vH+-ATPase in GTP-Gαo-mediated synaptic vesicle swelling. Results from this study, demonstrate that the GTP-Gαo-mediated vesicle swelling is vH+-ATPase–dependent, and pH sensitive. Zeta potential measurements of isolated synaptic vesicles further demonstrate, a bafilomycin-sensitive vesicle acidification, following the GTP-Gαo-induced swelling stimulus. Since water channels are bidirectional, and the vH+-ATPase inhibitor bafilomycin decreases both the volume of isolated synaptic vesicles and GTP-mastoparan stimulated swelling, suggests vH+-ATPase to be upstream of AQP-6, in the pathway leading from Gαo-stimulated swelling of synaptic vesicles. Vesicle acidification is therefore a prerequisite for AQP-6 mediated gating of water into synaptic vesicles. PMID:19610106

  15. Regulation of cytosolic pH and lactic acid release in mesangial cells overexpressing GLUT1.

    PubMed

    Lang, Karl S; Mueller, Matthias M; Tanneur, Valerie; Wallisch, Sabine; Fedorenko, Olga; Palmada, Monica; Lang, Florian; Bröer, Stefan; Heilig, Charles W; Schleicher, Erwin; Weigert, Cora

    2003-10-01

    Anaerobic glycolysis leads to the formation of lactate and H+ and thus imposes a significant challenge on cytosolic acid/base regulation. Cytosolic acidification, on the other hand, is known to inhibit flux through glycolysis and lactate formation. To explore the interplay of cytosolic pH and glycolysis, rat mesangial cells transfected with the glucose transporter GLUT1 (GLUT1 cells) were compared with those transfected with beta-galactosidase (LacZ cells). In the presence of extracellular glucose, the glycolytic rate was one order of magnitude higher in GLUT1 cells than in LacZ cells. Cytosolic pH (pHi) was significantly higher in GLUT1 than LacZ cells, an effect abolished in the presence of Na+/H+ exchange inhibitor ethylisopropylamiloride (1 micromol/L). Addition of 40 mmol/L lactate led to marked cytosolic acidification, which was in both cell types blunted by O-methyl-glucose (20 mmol/L) and completely abolished by 100 micromol/L phloretin and 1 mmol/L p-chloromercuribenzene-sulphonic acid (p-CMBS) and in LacZ cells only by glucose (20 mmol/L). The functional characterization points to the involvement of a lactic acid transporter from the monocarboxylate transporter (MCT) family, particularly MCT1. Reverse transcription-polymerase chain reaction (RT-PCR) indeed disclosed the expression of MCT1 and MCT2 in both GLUT1 and LacZ cells. Overexpression of GLUT1 leads to cytosolic alkalinization of mesangial cells depending on functional Na+/H+ exchanger but not on Na+ independent H+ transport.

  16. Release of canine parvovirus from endocytic vesicles.

    PubMed

    Suikkanen, Sanna; Antila, Mia; Jaatinen, Anne; Vihinen-Ranta, Maija; Vuento, Matti

    2003-11-25

    Canine parvovirus (CPV) is a small nonenveloped virus with a single-stranded DNA genome. CPV enters cells by clathrin-mediated endocytosis and requires an acidic endosomal step for productive infection. Virion contains a potential nuclear localization signal as well as a phospholipase A(2) like domain in N-terminus of VP1. In this study we characterized the role of PLA(2) activity on CPV entry process. PLA(2) activity of CPV capsids was triggered in vitro by heat or acidic pH. PLA(2) inhibitors inhibited the viral proliferation suggesting that PLA(2) activity is needed for productive infection. The N-terminus of VP1 was exposed during the entry, suggesting that PLA(2) activity might have a role during endocytic entry. The presence of drugs modifying endocytosis (amiloride, bafilomycin A(1), brefeldin A, and monensin) caused viral proteins to remain in endosomal/lysosomal vesicles, even though the drugs were not able to inhibit the exposure of VP1 N-terminal end. These results indicate that the exposure of N-terminus of VP1 alone is not sufficient to allow CPV to proliferate. Some other pH-dependent changes are needed for productive infection. In addition to blocking endocytic entry, amiloride was able to block some postendocytic steps. The ability of CPV to permeabilize endosomal membranes was demonstrated by feeding cells with differently sized rhodamine-conjugated dextrans together with the CPV in the presence or in the absence of amiloride, bafilomycin A(1), brefeldin A, or monensin. Dextran with a molecular weight of 3000 was released from vesicles after 8 h of infection, while dextran with a molecular weight of 10,000 was mainly retained in vesicles. The results suggest that CPV infection does not cause disruption of endosomal vesicles. However, the permeability of endosomal membranes apparently changes during CPV infection, probably due to the PLA(2) activity of the virus. These results suggest that parvoviral PLA(2) activity is essential for productive

  17. Lower pH values of weakly acidic refluxes as determinants of heartburn perception in gastroesophageal reflux disease patients with normal esophageal acid exposure.

    PubMed

    de Bortoli, N; Martinucci, I; Savarino, E; Franchi, R; Bertani, L; Russo, S; Ceccarelli, L; Costa, F; Bellini, M; Blandizzi, C; Savarino, V; Marchi, S

    2016-01-01

    Multichannel impedance pH monitoring has shown that weakly acidic refluxes are able to generate heartburn. However, data on the role of different pH values, ranging between 4 and 7, in the generation of them are lacking. The aim of this study was to evaluate whether different pH values of weakly acidic refluxes play a differential role in provoking reflux symptoms in endoscopy-negative patients with physiological esophageal acid exposure time and positive symptom index and symptom association probability for weakly acidic refluxes. One hundred and forty-three consecutive patients with gastroesophageal reflux disease, nonresponders to proton pump inhibitors (PPIs), were allowed a washout from PPIs before undergoing: upper endoscopy, esophageal manometry, and multichannel impedance pH monitoring. In patients with both symptom index and symptom association probability positive for weakly acidic reflux, each weakly acidic reflux was evaluated considering exact pH value, extension, physical characteristics, and correlation with heartburn. Forty-five patients with normal acid exposure time and positive symptom association probability for weakly acidic reflux were identified. The number of refluxes not heartburn related was higher than those heartburn related. In all distal and proximal liquid refluxes, as well as in distal mixed refluxes, the mean pH value of reflux events associated with heartburn was significantly lower than that not associated. This condition was not confirmed for proximal mixed refluxes. Overall, a low pH of weakly acidic reflux represents a determinant factor in provoking heartburn. This observation contributes to better understand the pathophysiology of symptoms generated by weakly acidic refluxes, paving the way toward the search for different therapeutic approaches to this peculiar condition of esophageal hypersensitivity.

  18. Ruminant Nutrition Symposium: Role of fermentation acid absorption in the regulation of ruminal pH.

    PubMed

    Aschenbach, J R; Penner, G B; Stumpff, F; Gäbel, G

    2011-04-01

    Highly fermentable diets are rapidly converted to organic acids [i.e., short-chain fatty acids (SCFA) and lactic acid] within the rumen. The resulting release of protons can constitute a challenge to the ruminal ecosystem and animal health. Health disturbances, resulting from acidogenic diets, are classified as subacute and acute acidosis based on the degree of ruminal pH depression. Although increased acid production is a nutritionally desired effect of increased concentrate feeding, the accumulation of protons in the rumen is not. Consequently, mechanisms of proton removal and their quantitative importance are of major interest. Saliva buffers (i.e., bicarbonate, phosphate) have long been identified as important mechanisms for ruminal proton removal. An even larger proportion of protons appears to be removed from the rumen by SCFA absorption across the ruminal epithelium, making efficiency of SCFA absorption a key determinant for the individual susceptibility to subacute ruminal acidosis. Proceeding initially from a model of exclusively diffusional absorption of fermentation acids, several protein-dependent mechanisms have been discovered over the last 2 decades. Although the molecular identity of these proteins is mostly uncertain, apical acetate absorption is mediated, to a major degree, via acetate-bicarbonate exchange in addition to another nitrate-sensitive, bicarbonate-independent transport mechanism and lipophilic diffusion. Propionate and butyrate also show partially bicarbonate-dependent transport modes. Basolateral efflux of SCFA and their metabolites has to be mediated primarily by proteins and probably involves the monocarboxylate transporter (MCT1) and anion channels. Although the ruminal epithelium removes a large fraction of protons from the rumen, it also recycles protons to the rumen via apical sodium-proton exchanger, NHE. The latter is stimulated by ruminal SCFA absorption and salivary Na(+) secretion and protects epithelial integrity. Finally

  19. Effect of pH alkaline salts of fatty acids on the inhibition of bacteria associated with poultry processing

    USDA-ARS?s Scientific Manuscript database

    The agar diffusion assay was used to examine the effect of pH on the ability of alkaline salts of three fatty acids (FA) to inhibit growth of bacteria associated with poultry processing. FA solutions were prepared by dissolving 0.5 M concentrations of caprylic, capric, or lauric acid in separate ali...

  20. Poking Vesicles

    NASA Astrophysics Data System (ADS)

    Powers, Thomas R.; Huber, Greg; Goldstein, Raymond E.

    2000-03-01

    Lipid vesicles can exhibit a variety of interesting shapes when subject to point forcing, as has been demonstrated with laser-tweezed beads and with growing microtubules. Using numerical and analytic techniques, we study the force vs. extension in two regimes. At low forces, the resistance to deformation is primarily entropic, with a tension given by the fluctuating bending modes. At high forces, the thermal wrinkles are smoothed out, and there is bending and stretching which balance in a thin tether, which we treat using boundary-layer techniques. This work was supported in part by the Harvard MRSEC and Army Research Office Grant DAA655-97-1-014 (TRP) and NSF DMR9812526 (GH & REG).

  1. Acid-extrusion from tissue: the interplay between membrane transporters and pH buffers.

    PubMed

    Hulikova, Alzbeta; Harris, Adrian L; Vaughan-Jones, Richard D; Swietach, Pawel

    2012-01-01

    The acid-base balance of cells is related to the concentration of free H⁺ ions. These are highly reactive, and their intracellular concentration must be regulated to avoid detrimental effects to the cell. H⁺ ion dynamics are influenced by binding to chelator substances ('buffering'), and by the production, diffusion and membrane-transport of free H⁺ ions or of the H⁺-bound chelators. Intracellular pH (pHi) regulation aims to balance this system of diffusion-reaction-transport processes at a favourable steady-state pHi. The ability of cells to regulate pHi may set a limit to tissue growth and can be subject to selection pressures. Cancer cells have been postulated to respond favourably to such selection pressures by evolving a better means of pHi regulation. A particularly important feature of tumour pHi regulation is acid-extrusion, which involves H⁺-extrusion and HCO₃⁻-uptake by membrane-bound transporter-proteins. Extracellular CO₂/HCO₃⁻ buffer facilitates these membrane-transport processes. As a mobile pH-buffer, CO₂/HCO₃⁻ protects the extracellular space from excessive acidification that could otherwise inhibit further acid-extrusion. CO₂/HCO₃⁻ also provides substrate for HCO₃⁻-transporters. However, the inherently slow reaction kinetics of CO₂/HCO₃⁻ can be rate-limiting for acid-extrusion. To circumvent this, cells can express extracellular-facing carbonic anhydrase enzymes to accelerate the attainment of equilibrium between CO₂, HCO₃⁻ and H⁺. The acid-extrusion apparatus has been proposed as a target for anti-cancer therapy. The major targets include H⁺ pumps, Na⁺/H⁺ exchangers and carbonic anhydrases. The effectiveness of such therapy will depend on the correct identification of rate-limiting steps in pHi regulation in a specific type of cancer.

  2. PARK9-associated ATP13A2 localizes to intracellular acidic vesicles and regulates cation homeostasis and neuronal integrity

    PubMed Central

    Ramonet, David; Podhajska, Agata; Stafa, Klodjan; Sonnay, Sarah; Trancikova, Alzbeta; Tsika, Elpida; Pletnikova, Olga; Troncoso, Juan C.; Glauser, Liliane; Moore, Darren J.

    2012-01-01

    Mutations in the ATP13A2 gene (PARK9, OMIM 610513) cause autosomal recessive, juvenile-onset Kufor-Rakeb syndrome and early-onset parkinsonism. ATP13A2 is an uncharacterized protein belonging to the P5-type ATPase subfamily that is predicted to regulate the membrane transport of cations. The physiological function of ATP13A2 in the mammalian brain is poorly understood. Here, we demonstrate that ATP13A2 is localized to intracellular acidic vesicular compartments in cultured neurons. In the human brain, ATP13A2 is localized to pyramidal neurons within the cerebral cortex and dopaminergic neurons of the substantia nigra. ATP13A2 protein levels are increased in nigral dopaminergic and cortical pyramidal neurons of Parkinson's disease brains compared with normal control brains. ATP13A2 levels are increased in cortical neurons bearing Lewy bodies (LBs) compared with neurons without LBs. Using short hairpin RNA-mediated silencing or overexpression to explore the function of ATP13A2, we find that modulating the expression of ATP13A2 reduces the neurite outgrowth of cultured midbrain dopaminergic neurons. We also find that silencing of ATP13A2 expression in cortical neurons alters the kinetics of intracellular pH in response to cadmium exposure. Furthermore, modulation of ATP13A2 expression leads to reduced intracellular calcium levels in cortical neurons. Finally, we demonstrate that silencing of ATP13A2 expression induces mitochondrial fragmentation in neurons. Oppositely, overexpression of ATP13A2 delays cadmium-induced mitochondrial fragmentation in neurons consistent with a neuroprotective effect. Collectively, this study reveals a number of intriguing neuronal phenotypes due to the loss- or gain-of-function of ATP13A2 that support a role for this protein in regulating intracellular cation homeostasis and neuronal integrity. PMID:22186024

  3. A photo Lewis acid generator (PhLAG): controlled photorelease of B(C6F5)3.

    PubMed

    Khalimon, Andrey Y; Piers, Warren E; Blackwell, James M; Michalak, David J; Parvez, Masood

    2012-06-13

    A molecule that releases the strong organometallic Lewis acid B(C(6)F(5))(3) upon irradiation with 254 nm light has been developed. This photo Lewis acid generator (PhLAG) now enables the photocontrolled initiation of several reactions catalyzed by this important Lewis acid. Herein is described the synthesis of the triphenylsulfonium salt of a carbamato borate based on a carbazole function, its establishment as a PhLAG, and the application of the photorelease of B(C(6)F(5))(3) to the fabrication of thin films of a polysiloxane material.

  4. Effect of pH and retention time on volatile fatty acids production during mixed culture fermentation.

    PubMed

    Jankowska, Ewelina; Chwiałkowska, Joanna; Stodolny, Mikołaj; Oleskowicz-Popiel, Piotr

    2015-08-01

    Mixed culture fermentation consists of stable microbial population hence waste could be potentially used as a substrates. The aim of the work was to investigate the impact of pH and retention time on the anaerobic mixed culture fermentation. Trials at different pH (4-12) in unbuffered systems were conducted for 5, 10 and 15days. The highest VFAs concentration was achieved after 15days at pH 10 (0.62g/gVSadded), promising results were also achieved for pH 11 (0.54g/gVSadded). For pH 4 and short retention time propionic acid was the major product instead of acetic acid. For batches run at 15days (besides pH 6) caproic acid presence was noticed whereas at pH 11 occurrence of succinic was quantified. Significant correlation between operational factors and fermentation's effluents was proved. Throughout changing simple operating parameters one could design process to produce desirable concentration and composition of VFAs.

  5. ATP- and growth substance-dependent cell-free enlargement of plasma membrane vesicles from soybean.

    PubMed

    Auderset, Guy; Morré, D James

    2006-01-01

    Growth hormone-responsive and nucleoside triphosphate-dependent enlargement of inside-out vesicles of plasma membranes from soybeans prepared by aqueous two-phase partition and everted by freezing and thawing has been achieved in a cell-free system. In the presence of 100 microM ATP in 40 mM HEPES buffer, pH 7, enlargement of isolated plasma membrane vesicles was accelerated by the synthetic plant growth factor, 2,4-dichlorophenoxyacetic acid (2,4-D), compared to ATP alone, 2,4-D alone or no additions. After 20 min with 1 microM 2,4-D, vesicles increased in diameter, 20% on average. Although vesicle diameters in the presence or absence of 2,4-D overlapped, the means were clearly separated. The 20% increases in diameter corresponded to a doubling of vesicle volume. Both 100 microM ATP and 1 microM 2,4-D were necessary to stimulate the cell-free vesicle enlargement. In the presence of 1 microM 2,4-D, enlargement observed with 100 microM ATP was greater than with either 10 microM ATP or 500 microM ATP alone. In the presence of 100 microM ATP, vesicle enlargement was proportional to the logarithm of 2,4-D concentration. With the growth-inactive 2,4-D analog, 2,3-D, no vesicle enlargement was observed either alone or in the presence of 100 microM ATP. Right side-out vesicles did not enlarge in response to either ATP, 2,4-D or the two in combination suggesting that the responsible ATP site was on the inside of the cell.

  6. Fucoxanthin from brown seaweed Sargassum cristaefolium tea in acid pH

    NASA Astrophysics Data System (ADS)

    Kartikaningsih, Hartati; Mufti, Eka Deviana; Nurhanief, Ardian Eko

    2017-05-01

    Dried tea Sargassum cristaefolium contains the pigment fucoxanthin, which is responsible for the red-orange color found in brown algae, and is a kind of photosynthetic pigment. Fucoxanthin can be used as an anti-obesity, anticancer, anti-cholesterol, and anti-diabetic agent and as a food colorant, but it is very unstable. The aim of this research was to determine the stability of fucoxanthin from dried tea brown algae at different pH (2, 6). This involved thin layer chromatography, peak absorption, wavelength analysis and reposition in FTIR. The research showed that fucoxanthin from fresh and dried tea Sargassum cristaefolium using chromatography columns had an orange color, Rf value of 0.26-0.28, and a spectral pattern in acetone solvent of 446.3-447.4 λmax. Fucoxanthin at pH 2 showed that there was no allenic group, as fucoxanthin solution had a pale yellow color. It is therefore shown that fucoxanthin is not stable in acid solution.

  7. Quadruplex structure of polyriboinosinic acid: dependence on alkali metal ion concentration, pH and temperature.

    PubMed

    Petrovic, Ana G; Polavarapu, Prasad L

    2008-02-21

    The vibrational infrared (IR) absorption and vibrational circular dichroism (VCD) spectral changes of polyinosinic acid (polyI) as a function of alkali metal ion concentration, temperature and pH have been investigated to establish how changes in spectral features relate to the structural modifications of polyI. A single positive VCD couplet associated with the carbonyl absorption band is considered to be the signature of quadruplex structure for polyI. The disruption of the quadruplex structure with temperature increase or pH increase at low alkali metal ion concentration is evidenced by the disappearance of this positive VCD couplet. The absence of any VCD signal upon quadruplex disruption indicates that the newly formed structure lacks helical chirality and is likely to be disordered. In the presence of 1 M NaCl or 0.1 M NaCl, the heat-induced quadruplex disruption is completely reversible. A mildly alkaline environment, in the presence of 0.1 M NaCl, is not sufficient to support the quadruplex structure of polyI. Trehalose-assisted polyI film at room temperature exhibits the same quadruplex spectral signature as that seen for solution at room temperature, but the quadruplex spectral signature in the film state remains at higher temperature, unlike in solution. This indicates that the quadruplex structure of polyI in the film state resists heat-induced disruptions.

  8. Aerosol pH buffering in the southeastern US: Fine particles remain highly acidic despite large reductions in sulfate

    NASA Astrophysics Data System (ADS)

    Weber, R. J.; Guo, H.; Russell, A. G.; Nenes, A.

    2015-12-01

    pH is a critical aerosol property that impacts many atmospheric processes, including biogenic secondary organic aerosol formation, gas-particle phase partitioning, and mineral dust or redox metal mobilization. Particle pH has also been linked to adverse health effects. Using a comprehensive data set from the Southern Oxidant and Aerosol Study (SOAS) as the basis for thermodynamic modeling, we have shown that particles are currently highly acidic in the southeastern US, with pH between 0 and 2. Sulfate and ammonium are the main acid-base components that determine particle pH in this region, however they have different sources and their concentrations are changing. Over 15 years of network data show that sulfur dioxide emission reductions have resulted in a roughly 70 percent decrease in sulfate, whereas ammonia emissions, mainly link to agricultural activities, have been largely steady, as have gas phase ammonia concentrations. This has led to the view that particles are becoming more neutralized. However, sensitivity analysis, based on thermodynamic modeling, to changing sulfate concentrations indicates that particles have remained highly acidic over the past decade, despite the large reductions in sulfate. Furthermore, anticipated continued reductions of sulfate and relatively constant ammonia emissions into the future will not significantly change particle pH until sulfate drops to clean continental background levels. The result reshapes our expectation of future particle pH and implies that atmospheric processes and adverse health effects linked to particle acidity will remain unchanged for some time into the future.

  9. A single pH fluorescent probe for biosensing and imaging of extreme acidity and extreme alkalinity.

    PubMed

    Chao, Jian-Bin; Wang, Hui-Juan; Zhang, Yong-Bin; Li, Zhi-Qing; Liu, Yu-Hong; Huo, Fang-Jun; Yin, Cai-Xia; Shi, Ya-Wei; Wang, Juan-Juan

    2017-07-04

    A simple tailor-made pH fluorescent probe 2-benzothiazole (N-ethylcarbazole-3-yl) hydrazone (Probe) is facilely synthesized by the condensation reaction of 2-hydrazinobenzothiazole with N-ethylcarbazole-3-formaldehyde, which is a useful fluorescent probe for monitoring extremely acidic and alkaline pH, quantitatively. The pH titrations indicate that Probe displays a remarkable emission enhancement with a pKa of 2.73 and responds linearly to minor pH fluctuations within the extremely acidic range of 2.21-3.30. Interestingly, Probe also exhibits strong pH-dependent characteristics with pKa 11.28 and linear response to extreme-alkalinity range of 10.41-12.43. In addition, Probe shows a large Stokes shift of 84 nm under extremely acidic and alkaline conditions, high selectivity, excellent sensitivity, good water-solubility and fine stability, all of which are favorable for intracellular pH imaging. The probe is further successfully applied to image extremely acidic and alkaline pH values fluctuations in E. coli cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Direct detection of acidity, alkalinity, and pH with membrane electrodes.

    PubMed

    Crespo, Gastón A; Ghahraman Afshar, Majid; Bakker, Eric

    2012-12-04

    An electrochemical sensing protocol based on supported liquid ion-selective membranes for the direct detection of total alkalinity of a sample that contains a weak base such as Tris (pK(a) = 8.2) is presented here for the first time. Alkalinity is determined by imposing a defined flux of hydrogen ions from the membrane to the sample with an applied current. The transition time at which the base species at the membrane-sample interface depletes owing to diffusion limitation is related to sample alkalinity in this chronopotentiometric detection mode. The same membrane is shown to detect pH (by zero current potentiometry) and acidity and alkalinity (by chronopotentiometry at different current polarity). This principle may become a welcome tool for the in situ determination of these characteristics in complex samples such as natural waters.

  11. Synaptic vesicles contain small ribonucleic acids (sRNAs) including transfer RNA fragments (trfRNA) and microRNAs (miRNA)

    PubMed Central

    Li, Huinan; Wu, Cheng; Aramayo, Rodolfo; Sachs, Matthew S.; Harlow, Mark L.

    2015-01-01

    Synaptic vesicles (SVs) are neuronal presynaptic organelles that load and release neurotransmitter at chemical synapses. In addition to classic neurotransmitters, we have found that synaptic vesicles isolated from the electric organ of Torpedo californica, a model cholinergic synapse, contain small ribonucleic acids (sRNAs), primarily the 5′ ends of transfer RNAs (tRNAs) termed tRNA fragments (trfRNAs). To test the evolutionary conservation of SV sRNAs we examined isolated SVs from the mouse central nervous system (CNS). We found abundant levels of sRNAs in mouse SVs, including trfRNAs and micro RNAs (miRNAs) known to be involved in transcriptional and translational regulation. This discovery suggests that, in addition to inducing changes in local dendritic excitability through the release of neurotransmitters, SVs may, through the release of specific trfRNAs and miRNAs, directly regulate local protein synthesis. We believe these findings have broad implications for the study of chemical synaptic transmission. PMID:26446566

  12. Glucose-Responsive Supramolecular Vesicles Based on Water-Soluble Pillar[5]arene and Pyridylboronic Acid Derivative for Controlled Insulin Delivery.

    PubMed

    Gao, Lei; Wang, Tingting; Jia, Keke; Wu, Xuan; Yao, Chenhao; Shao, Wei; Zhang, Dongmei; Hu, Xiao-Yu; Wang, Leyong

    2017-03-22

    The stimuli-responsive behavior of supramolecular nanocarriers is crucial for their potential applications as smart drug delivery systems. We hereby constructed a glucose-responsive supramolecular drug delivery system constructed by the host-guest interaction between a water-soluble pillar[5]arene (WP5) and a pyridylboronic acid derivative (G) for insulin delivery and controlled release at physiological condition, which represents the ideal treatment of diabetes mellitus. The drug loading and in vitro drug release experiments demonstrated that large molecular weight insulin could be successfully encapsulated into the vesicles with high loading efficiency, which to the best of our knowledge, is the first example of small size supramolecular vesicles with excellent encapsulation capacity of large protein molecule. Moreover, FITC-labeled insulin was used to evaluate the release behavior of insulin, and it was demonstrated that high glucose concentration could facilitate the quick release of insulin, suggesting a smart DDS for the potential application in controlled insulin release only under hyperglycemic conditions. Finally, we demonstrated that these supramolecular nanocarriers have good cytocompatibility, which is essential for their further biomedical applications. The present study provides a novel strategy for the construction of glucose-responsive smart supramolecular drug delivery system, which has potential applications for the treatment of diabetes mellitus.

  13. Synaptic vesicles contain small ribonucleic acids (sRNAs) including transfer RNA fragments (trfRNA) and microRNAs (miRNA).

    PubMed

    Li, Huinan; Wu, Cheng; Aramayo, Rodolfo; Sachs, Matthew S; Harlow, Mark L

    2015-10-08

    Synaptic vesicles (SVs) are neuronal presynaptic organelles that load and release neurotransmitter at chemical synapses. In addition to classic neurotransmitters, we have found that synaptic vesicles isolated from the electric organ of Torpedo californica, a model cholinergic synapse, contain small ribonucleic acids (sRNAs), primarily the 5' ends of transfer RNAs (tRNAs) termed tRNA fragments (trfRNAs). To test the evolutionary conservation of SV sRNAs we examined isolated SVs from the mouse central nervous system (CNS). We found abundant levels of sRNAs in mouse SVs, including trfRNAs and micro RNAs (miRNAs) known to be involved in transcriptional and translational regulation. This discovery suggests that, in addition to inducing changes in local dendritic excitability through the release of neurotransmitters, SVs may, through the release of specific trfRNAs and miRNAs, directly regulate local protein synthesis. We believe these findings have broad implications for the study of chemical synaptic transmission.

  14. Infectious pancreatic necrosis virus in fish by-products is inactivated with inorganic acid (pH 1) and base (pH 12).

    PubMed

    Myrmel, M; Modahl, I; Nygaard, H; Lie, K M

    2014-04-01

    The aquaculture industry needs a simple, inexpensive and safe method for the treatment of fish waste without heat. Microbial inactivation by inorganic acid (HCl) or base (KOH) was determined using infectious pancreatic necrosis virus (IPNV) as a model organism for fish pathogens. Salmonella and spores of Clostridium perfringens were general hygiene indicators in supplementary examinations. IPNV, which is considered to be among the most chemical- and heat-resistant fish pathogens, was reduced by more than 3 log in 4 h at pH 1.0 and pH 12.0. Salmonella was rapidly inactivated by the same treatment, whereas spores of C. perfringens were hardly affected. The results indicate that low and high pH treatment could be particularly suitable for fish waste destined for biogas production. pH treatment at aquaculture production sites could reduce the spread of fish pathogens during storage and transportation without disturbing the anaerobic digestion process. The treatment could also be an alternative to the current energy-intensive steam pressure sterilization of fish waste to be used by the bioenergy, fertilizer and soil improver industries. © 2013 John Wiley & Sons Ltd.

  15. Extracellular Vesicles in Renal Pathophysiology

    PubMed Central

    Pomatto, Margherita A. C.; Gai, Chiara; Bussolati, Benedetta; Camussi, Giovanni

    2017-01-01

    Extracellular vesicles are a heterogeneous population of microparticles released by virtually all living cells which have been recently widely investigated in different biological fields. They are typically composed of two primary types (exosomes and microvesicles) and are recently commanding increasing attention as mediators of cellular signaling. Indeed, these vesicles can affect recipient cells by carrying and delivering complex cargos of biomolecules (including proteins, lipids and nucleic acids), protected from enzymatic degradation in the environment. Their importance has been demonstrated in the pathophysiology of several organs, in particular in kidney, where different cell types secrete extracellular vesicles that mediate their communication with downstream urinary tract cells. Over the past few years, evidence has been shown that vesicles participate in kidney development and normal physiology. Moreover, EVs are widely demonstrated to be implicated in cellular signaling during renal regenerative and pathological processes. Although many EV mechanisms are still poorly understood, in particular in kidney, the discovery of their role could help to shed light on renal biological processes which are so far elusive. Lastly, extracellular vesicles secreted by renal cells gather in urine, thus becoming a great resource for disease or recovery markers and a promising non-invasive diagnostic instrument for renal disease. In the present review, we discuss the most recent findings on the role of extracellular vesicles in renal physiopathology and their potential implication in diagnosis and therapy. PMID:28638822

  16. Extracellular Vesicles in Renal Pathophysiology.

    PubMed

    Pomatto, Margherita A C; Gai, Chiara; Bussolati, Benedetta; Camussi, Giovanni

    2017-01-01

    Extracellular vesicles are a heterogeneous population of microparticles released by virtually all living cells which have been recently widely investigated in different biological fields. They are typically composed of two primary types (exosomes and microvesicles) and are recently commanding increasing attention as mediators of cellular signaling. Indeed, these vesicles can affect recipient cells by carrying and delivering complex cargos of biomolecules (including proteins, lipids and nucleic acids), protected from enzymatic degradation in the environment. Their importance has been demonstrated in the pathophysiology of several organs, in particular in kidney, where different cell types secrete extracellular vesicles that mediate their communication with downstream urinary tract cells. Over the past few years, evidence has been shown that vesicles participate in kidney development and normal physiology. Moreover, EVs are widely demonstrated to be implicated in cellular signaling during renal regenerative and pathological processes. Although many EV mechanisms are still poorly understood, in particular in kidney, the discovery of their role could help to shed light on renal biological processes which are so far elusive. Lastly, extracellular vesicles secreted by renal cells gather in urine, thus becoming a great resource for disease or recovery markers and a promising non-invasive diagnostic instrument for renal disease. In the present review, we discuss the most recent findings on the role of extracellular vesicles in renal physiopathology and their potential implication in diagnosis and therapy.

  17. Nitrification in a Biofilm at Low pH Values: Role of In Situ Microenvironments and Acid Tolerance

    PubMed Central

    Gieseke, Armin; Tarre, Sheldon; Green, Michal; de Beer, Dirk

    2006-01-01

    The sensitivity of nitrifying bacteria to acidic conditions is a well-known phenomenon and generally attributed to the lack and/or toxicity of substrates (NH3 and HNO2) with decreasing pHs. In contrast, we observed strong nitrification at a pH around 4 in biofilms grown on chalk particles and investigated the following hypotheses: the presence of less acidic microenvironments and/or the existence of acid-tolerant nitrifiers. Microelectrode measurements (in situ and under various experimental conditions) showed no evidence of a neutral microenvironment, either within the highly active biofilm colonizing the chalk surface or within a control biofilm grown on a nonbuffering (i.e., sintered glass) surface under acidic pH. A 16S rRNA approach (clone libraries and fluorescence in situ hybridizations) did not reveal uncommon nitrifying (potentially acid-tolerant) strains. Instead, we found a strongly acidic microenvironment, evidence for a clear adaptation to the low pH in situ, and the presence of nitrifying populations related to subgroups with low Kms for ammonia (Nitrosopira spp., Nitrosomonas oligotropha, and Nitrospira spp.). Acid-consuming (chalk dissolution) and acid-producing (ammonia oxidation) processes are equilibrated on a low-pH steady state that is controlled by mass transfer limitation through the biofilm. Strong affinity to ammonia and possibly the expression of additional functions, e.g., ammonium transporters, are adaptations that allow nitrifiers to cope with acidic conditions in biofilms and other habitats. PMID:16751543

  18. Nitrification in a biofilm at low pH values: role of in situ microenvironments and acid tolerance.

    PubMed

    Gieseke, Armin; Tarre, Sheldon; Green, Michal; de Beer, Dirk

    2006-06-01

    The sensitivity of nitrifying bacteria to acidic conditions is a well-known phenomenon and generally attributed to the lack and/or toxicity of substrates (NH3 and HNO2) with decreasing pHs. In contrast, we observed strong nitrification at a pH around 4 in biofilms grown on chalk particles and investigated the following hypotheses: the presence of less acidic microenvironments and/or the existence of acid-tolerant nitrifiers. Microelectrode measurements (in situ and under various experimental conditions) showed no evidence of a neutral microenvironment, either within the highly active biofilm colonizing the chalk surface or within a control biofilm grown on a nonbuffering (i.e., sintered glass) surface under acidic pH. A 16S rRNA approach (clone libraries and fluorescence in situ hybridizations) did not reveal uncommon nitrifying (potentially acid-tolerant) strains. Instead, we found a strongly acidic microenvironment, evidence for a clear adaptation to the low pH in situ, and the presence of nitrifying populations related to subgroups with low Km s for ammonia (Nitrosopira spp., Nitrosomonas oligotropha, and Nitrospira spp.). Acid-consuming (chalk dissolution) and acid-producing (ammonia oxidation) processes are equilibrated on a low-pH steady state that is controlled by mass transfer limitation through the biofilm. Strong affinity to ammonia and possibly the expression of additional functions, e.g., ammonium transporters, are adaptations that allow nitrifiers to cope with acidic conditions in biofilms and other habitats.

  19. Alteration of the phospho- or neutral lipid content and fatty acid composition in Listeria monocytogenes due to acid adaptation mechanisms for hydrochloric, acetic and lactic acids at pH 5.5 or benzoic acid at neutral pH.

    PubMed

    Mastronicolis, Sofia K; Berberi, Anita; Diakogiannis, Ioannis; Petrova, Evanthia; Kiaki, Irene; Baltzi, Triantafillia; Xenikakis, Polydoros

    2010-10-01

    This study provides a first approach to observe the effects on Listeria monocytogenes of cellular exposure to acid stress at low or neutral pH, notably how phospho- or neutral lipids are involved in this mechanism, besides the fatty acid profile alteration. A thorough investigation of the composition of polar and neutral lipids from L. monocytogenes grown at pH 5.5 in presence of hydrochloric, acetic and lactic acids, or at neutral pH 7.3 in presence of benzoic acid, is described relative to cells grown in acid-free medium. The results showed that only low pH values enhance the antimicrobial activity of an acid. We suggest that, irrespective of pH, the acid adaptation response will lead to a similar alteration in fatty acid composition [decreasing the ratio of branched chain/saturated straight fatty acids of total lipids], mainly originating from the neutral lipid class of adapted cultures. Acid adaptation in L. monocytogenes was correlated with a decrease in total lipid phosphorus and, with the exception of cells adapted to benzoic acid, this change in the amount of phosphorus reflected a higher content of the neutral lipid class. Upon acetic or benzoic acid stress the lipid phosphorus proportion was analysed in the main phospholipids present: cardiolipin, phosphatidylglycerol, phosphoaminolipid and phosphatidylinositol. Interestingly only benzoic acid had a dramatic effect on the relative quantities of these four phospholipids.

  20. Alteration of the phospho- or neutral lipid content and fatty acid composition in Listeria monocytogenes due to acid adaptation mechanisms for hydrochloric, acetic and lactic acids at pH 5.5 or benzoic acid at neutral pH

    PubMed Central

    Berberi, Anita; Diakogiannis, Ioannis; Petrova, Evanthia; Kiaki, Irene; Baltzi, Triantafillia; Xenikakis, Polydoros

    2010-01-01

    This study provides a first approach to observe the effects on Listeria monocytogenes of cellular exposure to acid stress at low or neutral pH, notably how phospho- or neutral lipids are involved in this mechanism, besides the fatty acid profile alteration. A thorough investigation of the composition of polar and neutral lipids from L. monocytogenes grown at pH 5.5 in presence of hydrochloric, acetic and lactic acids, or at neutral pH 7.3 in presence of benzoic acid, is described relative to cells grown in acid-free medium. The results showed that only low pH values enhance the antimicrobial activity of an acid. We suggest that, irrespective of pH, the acid adaptation response will lead to a similar alteration in fatty acid composition [decreasing the ratio of branched chain/saturated straight fatty acids of total lipids], mainly originating from the neutral lipid class of adapted cultures. Acid adaptation in L. monocytogenes was correlated with a decrease in total lipid phosphorus and, with the exception of cells adapted to benzoic acid, this change in the amount of phosphorus reflected a higher content of the neutral lipid class. Upon acetic or benzoic acid stress the lipid phosphorus proportion was analysed in the main phospholipids present: cardiolipin, phosphatidylglycerol, phosphoaminolipid and phosphatidylinositol. Interestingly only benzoic acid had a dramatic effect on the relative quantities of these four phospholipids. PMID:20379849

  1. Oxidative interactions between the erythrocyte membrane and phosphatidylcholine vesicles.

    PubMed

    Yang, E; Huestis, W H

    1994-05-20

    Sonicated unilamellar phosphatidylcholine vesicles induce hemoglobin oxidation in erythrocytes and resealed membrane fragments (buds) at pH 5.5. No such oxidation was observed in vesicle-bud mixtures at pH 7.4, in cells or buds suspended in pH 5.5 buffer, or in cells incubated with multilamellar lipid vesicles at pH 5.5. In buds, vesicle-induced hemoglobin oxidation was accompanied by lipid peroxidation and formation of covalent high molecular weight protein aggregates. The causative relationships among the oxidative events were examined using selective antioxidants and membrane fragments in which the cytoplasmic domain of band 3 was cleaved. Protein cross-linking and lipid peroxidation were found to be independent events, but both were found only in concert with heme oxidation. Further, vesicle-induced hemoglobin oxidation was found to be correlated with quasi-stable adsorption of intact vesicles to cells; intercalation of foreign lipid into the cell bilayer was not required. The inability of multilamellar vesicles to induce these low pH oxidative effects suggests a steric limitation on this cell-vesicle association. The results suggest that membrane component reorganization and patching induced by low pH may enhance vesicle adsorption, which in turn initiates oxidative damage.

  2. Influence of acidic pH on keratinocyte function and re-epithelialisation of human in vitro wounds.

    PubMed

    Lönnqvist, Susanna; Emanuelsson, Peter; Kratz, Gunnar

    2015-01-01

    Chronic wounds are one of the greatest challenges for the healthcare system. Today, a plethora of dressings are used in the treatment of these wounds, each with specific influence on the wound environment. Due to differences in the permeability of the dressings the use will result in differences in the pH balance in the wound bed. However, little is known about how changes in the pH in the wound environment affect the different phases of the healing process. The aim of the present study was to investigate the effects of acidic pH on the regeneration phase by studying keratinocyte function in vitro and re-epithelialisation in an in vitro model of human skin. In vitro assays showed reduced viability and migration rates in human keratinocytes when pH was lowered. Real time PCR revealed differential expression of genes related to wound healing and environmental impairment. Tissue culture showed no re-epithelialisation of wounds subjected to pH 5.0 and moderate re-epithelialisation at pH 6.0, compared to controls at pH 7.4. The results indicate that lowering pH down to pH 5.0 in wounds is counterproductive in aspect of keratinocyte function which is crucial for successful wound healing.

  3. Reactive solute transport in an acidic stream: Experimental pH increase and simulation of controls on pH, aluminum, and iron

    USGS Publications Warehouse

    Broshears, R.E.; Runkel, R.L.; Kimball, B.A.; McKnight, Diane M.; Bencala, K.E.

    1996-01-01

    Solute transport simulations quantitatively constrained hydrologic and geochemical hypotheses about field observations of a pH modification in an acid mine drainage stream. Carbonate chemistry, the formation of solid phases, and buffering interactions with the stream bed were important factors in explaining the behavior of pH, aluminum, and iron. The precipitation of microcrystalline gibbsite accounted for the behavior of aluminum; precipitation of Fe(OH)3 explained the general pattern of iron solubility. The dynamic experiment revealed limitations on assumptions that reactions were controlled only by equilibrium chemistry. Temporal variation in relative rates of photoreduction and oxidation influenced iron behavior. Kinetic limitations on ferrous iron oxidation and hydrous oxide precipitation and the effects of these limitations on field filtration were evident. Kinetic restraints also characterized interaction between the water column and the stream bed, including sorption and desorption of protons from iron oxides at the sediment-water interface and post-injection dissolution of the precipitated aluminum solid phase.

  4. Feruloyl Dioleoyglycerol Antioxidant Capacity in Phospholipid Vesicles

    USDA-ARS?s Scientific Manuscript database

    Ferulic acid and its esters are known to be effective antioxidants. Feruloyl dioleoylglycerol was assessed for its ability to serve as an antioxidant in model membrane phospholipid vesicles. The molecule was incorporated into single-lamellar vesicles of 1,2-dioleoyl-sn-glycero-3-phosphocholine at ...

  5. Elucidation of insulin assembly at acidic and neutral pH: Characterization of low molecular weight oligomers.

    PubMed

    Mawhinney, Matthew T; Williams, Thomas L; Hart, James L; Taheri, Mitra L; Urbanc, Brigita

    2017-08-10

    Deficiency in insulin secretion and function that characterize type 2 diabetes often requires administration of extraneous insulin, leading to injection-site amyloidosis. Insulin aggregation at neutral pH is not well understood. Although oligomer formation is believed to play an important role, insulin oligomers have not been fully characterized yet. Here, we elucidate similarities and differences between in vitro insulin aggregation at acidic and neutral pH for a range of insulin concentrations (2.5-100 μM) by using kinetic thioflavin T fluorescence, circular dichroism, atomic force and electron microscopy imaging. Importantly, we characterize the size distribution of insulin oligomers at different assembly stages by the application of covalent cross-linking and gel electrophoresis. Our results show that at the earliest assembly stage, oligomers comprise up to 40% and 70% of soluble insulin at acidic and neutral pH, respectively. While the highest oligomer order increases with insulin concentration at acidic pH, the opposite tendency is observed at neutral pH, where oligomers up to heptamers are formed in 10 μM insulin. These findings suggest that oligomers may be on- and off-pathway assemblies for insulin at acidic and neutral pH, respectively. Agitation, which is required to induce insulin aggregation at neutral pH, is shown to increase fibril formation rate and fibrillar mass both by an order of magnitude. Insulin incubated under agitated conditions at neutral pH rapidly aggregates into large micrometer-sized aggregates, which may be of physiological relevance and provides insight into injection-site amyloidosis and toxic pulmonary aggregates induced by administration of extraneous insulin. © 2017 Wiley Periodicals, Inc.

  6. Boronate-Phenolic Network Capsules with Dual Response to Acidic pH and cis-Diols.

    PubMed

    Guo, Junling; Sun, Huanli; Alt, Karen; Tardy, Blaise L; Richardson, Joseph J; Suma, Tomoya; Ejima, Hirotaka; Cui, Jiwei; Hagemeyer, Christoph E; Caruso, Frank

    2015-08-26

    Dual-responsive boronate-phenolic network (BPN) capsules are fabricated by the complexation of phenylborate and phenolic materials. The BPN capsules are stable in the presence of competing carbohydrates, but dissociate at acidic pH or in the presence of competing cis-diols at physiological pH. This engineered capsule system provides a platform for a wide range of biological and biomedical applications. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. The effect of antacid on salivary pH in patients with and without dental erosion after multiple acid challenges.

    PubMed

    Dhuhair, Sarah; Dennison, Joseph B; Yaman, Peter; Neiva, Gisele F

    2015-04-01

    To evaluate the effect of antacid swish in the salivary pH values and to monitor the pH changes in subjects with and without dental erosion after multiple acid challenge tests. 20 subjects with tooth erosion were matched in age and gender with 20 healthy controls according to specific inclusion/exclusion criteria. Baseline measures were taken of salivary pH, buffering capacity and salivary flow rate using the Saliva Check System. Subjects swished with Diet Pepsi three times at 10-minute intervals. Changes in pH were monitored using a digital pH meter at 0-, 5-, and 10- minute intervals and at every 5 minutes after the third swish until pH resumed baseline value or 45 minutes relapse. Swishing regimen was repeated on a second visit, followed by swishing with sugar-free liquid antacid (Mylanta Supreme). Recovery times were also recorded. Data was analyzed using independent t-tests, repeated measures ANOVA, and Fisher's exact test (α= 0.05). Baseline buffering capacity and flow rate were not significantly different between groups (P= 0.542; P= 0.2831, respectively). Baseline salivary pH values were similar between groups (P= 0.721). No significant differences in salivary pH values were found between erosion and non-erosion groups in response to multiple acid challenges (P= 0.695) or antacid neutralization (P= 0.861). Analysis of salivary pH recovery time revealed no significant differences between groups after acid challenges (P= 0.091) or after the use of antacid (P= 0.118). There was a highly significant difference in the survival curves of the two groups on Day 2, with the non-erosion group resolving significantly faster than the erosion group (P= 0.0086).

  8. A water-soluble rhodamine B-derived fluorescent probe for pH monitoring and imaging in acidic regions

    NASA Astrophysics Data System (ADS)

    Cui, Peng; Jiang, Xuekai; Sun, Junyong; Zhang, Qiang; Gao, Feng

    2017-06-01

    A structurally simple, water-soluble rhodamine-derivatived fluorescent probe, which is responsive to acidic pH, was conveniently synthesized via a one-step condensation reaction of rhodamine B hydrazide and 4-formybenzene-1,3-disulfonate. As a stable and highly sensitive pH sensor, the probe displays an approximately 50-fold fluorescence enhancement over the pH range of 7.16-4.89 as the structure of probe changes from spirocyclic (weak fluorescent) to ring-open (strong fluorescent) with decreasing pH. The synthesized fluorescent probe is applied to the detection of pH changes in vitro and in vivo bioimaging of immortalized gastric cancer cells, with satisfactory results.

  9. Effect of pH on Fenton process using estimation of hydroxyl radical with salicylic acid as trapping reagent.

    PubMed

    Chang, Chen-Yu; Hsieh, Yung-Hsu; Cheng, Kai-Yuan; Hsieh, Ling-Ling; Cheng, Ta-Chih; Yao, Kuo-Shan

    2008-01-01

    This study estimates the yield of hydroxyl radical using salicylic acid as the trapping reagent and investigates the relationship between hydroxyl radical and pH value. The formation and variation of hydroxyl radical under different pH values were evaluated using reaction products, 2,3-DHBA, 2,5-DHBA, and catechol. The formation rate of hydroxyl radical was dependent on the ratio of ferrous ion to hydrogen peroxide and pH values. The difference between various pH values was explored. The kinetics and mechanisms of hydroxyl radical reactions were established in the Fenton process. Experimental results showed that the best reaction conditions were 8.5 mM H(2)O(2), 1.25 mM Fe(2 + ), Fe(2 + )/H(2)O(2) = 0.147 at pH 3 and the formation rate constant of hydroxyl radical was 1.12 x 10(11) M(-1) s(-1).

  10. First-principles calculation of thermodynamic stability of acids and bases under pH environment: A microscopic pH theory

    NASA Astrophysics Data System (ADS)

    Kim, Yong-Hyun; Kim, Kwiseon; Zhang, S. B.

    2012-04-01

    Despite being one of the most important thermodynamic variables, pH has yet to be incorporated into first-principles thermodynamics to calculate stability of acidic and basic solutes in aqueous solutions. By treating the solutes as defects in homogeneous liquids, we formulate a first-principles approach to calculate their formation energies under proton chemical potential, or pH, based on explicit molecular dynamics. The method draws analogy to first-principle calculations of defect formation energies under electron chemical potential, or Fermi energy, in semiconductors. From this, we propose a simple pictorial representation of the general theory of acid-base chemistry. By performing first-principles molecular dynamics of liquid water models with solutes, we apply the formulation to calculate formation energies of various neutral and charged solutes such as H+, OH-, NH3, NH4+, HCOOH, and HCOO- in water. The deduced auto-dissociation constant of water and the difference in the pKa values of NH3 and HCOOH show good agreement with known experimental values. Our first-principles approach can be further extended and applied to other bio- and electro-chemical molecules such as amino acids and redox reaction couples that could exist in aqueous environments to understand their thermodynamic stability.

  11. First-Principles Calculation of Thermodynamic Stability of Acids and Bases under pH Environment: A Microscopic pH Theory

    SciTech Connect

    Kim, Y. H.; Kim, K.; Zhang, S. B.

    2012-04-07

    Despite being one of the most important thermodynamic variables, pH has yet to be incorporated into first-principles thermodynamics to calculate stability of acidic and basic solutes in aqueous solutions. By treating the solutes as defects in homogeneous liquids, we formulate a first-principles approach to calculate their formation energies under proton chemical potential, or pH, based on explicit molecular dynamics. The method draws analogy to first-principle calculations of defect formation energies under electron chemical potential, or Fermi energy, in semiconductors. From this, we propose a simple pictorial representation of the general theory of acid-base chemistry. By performing first-principles molecular dynamics of liquid water models with solutes, we apply the formulation to calculate formation energies of various neutral and charged solutes such as H{sup +}, OH{sup -}, NH{sub 3}, NH{sub 4}{sup +}, HCOOH, and HCOO{sup -} in water. The deduced auto-dissociation constant of water and the difference in the pKa values of NH{sub 3} and HCOOH show good agreement with known experimental values. Our first-principles approach can be further extended and applied to other bio- and electro-chemical molecules such as amino acids and redox reaction couples that could exist in aqueous environments to understand their thermodynamic stability.

  12. First-principles calculation of thermodynamic stability of acids and bases under pH environment: a microscopic pH theory.

    PubMed

    Kim, Yong-Hyun; Kim, Kwiseon; Zhang, S B

    2012-04-07

    Despite being one of the most important thermodynamic variables, pH has yet to be incorporated into first-principles thermodynamics to calculate stability of acidic and basic solutes in aqueous solutions. By treating the solutes as defects in homogeneous liquids, we formulate a first-principles approach to calculate their formation energies under proton chemical potential, or pH, based on explicit molecular dynamics. The method draws analogy to first-principle calculations of defect formation energies under electron chemical potential, or Fermi energy, in semiconductors. From this, we propose a simple pictorial representation of the general theory of acid-base chemistry. By performing first-principles molecular dynamics of liquid water models with solutes, we apply the formulation to calculate formation energies of various neutral and charged solutes such as H(+), OH(-), NH(3), NH(4)(+), HCOOH, and HCOO(-) in water. The deduced auto-dissociation constant of water and the difference in the pKa values of NH(3) and HCOOH show good agreement with known experimental values. Our first-principles approach can be further extended and applied to other bio- and electro-chemical molecules such as amino acids and redox reaction couples that could exist in aqueous environments to understand their thermodynamic stability.

  13. Lactic acid fermentation from food waste with indigenous microbiota: Effects of pH, temperature and high OLR.

    PubMed

    Tang, Jialing; Wang, Xiaochang; Hu, Yisong; Zhang, Yongmei; Li, Yuyou

    2016-06-01

    The effects of pH, temperature and high organic loading rate (OLR) on lactic acid production from food waste without extra inoculum addition were investigated in this study. Using batch experiments, the results showed that although the hydrolysis rate increased with pH adjustment, the lactic acid concentration and productivity were highest at pH 6. High temperatures were suitable for solubilization but seriously restricted the acidification processes. The highest lactic acid yield (0.46g/g-TS) and productivity (278.1mg/Lh) were obtained at 37°C and pH 6. In addition, the lactic acid concentration gradually increased with the increase in OLR, and the semi-continuous reactor could be stably operated at an OLR of 18g-TS/Ld. However, system instability, low lactic acid yield and a decrease in VS removal were noticed at high OLRs (22g-TS/Ld). The concentrations of volatile fatty acids (VFAs) in the fermentation mixture were relatively low but slightly increased with OLR, and acetate was the predominant VFA component. Using high-throughput pyrosequencing, Lactobacillus from the raw food waste was found to selectively accumulate and become dominant in the semi-continuous reactor. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. The Relationship between Acidic Urinary pH and Overactive Bladder; Alkalization of Urine Improves the Symptoms of Overactive Bladder.

    PubMed

    Demirbas, Arif; Sarici, Hasmet; Kilinc, M Fatih; Telli, Onur; Ozgur, Berat Cem; Doluoglu, Omer Gokhan; Bozkurt, Selen

    2015-01-01

    To determine association between urine pH and OAB symptoms and to determine if urinary alkalization improves overactive bladder (OAB) symptoms. 659 patients with OAB were enrolled in this study between June 2012 and May 2014. 329 patients (group 1) were included in the final analysis. 201 adults were used as a control group (group 2). 24-hour urinary pH and the validated Turkish version of the OAB-V8 questionnaire were performed in patients. A 24-hour urine pH <6.2 was considered acidic urine. In the second part, a diet program was performed for 4 weeks in 30 participants. Urine pH values and OAB-V8 scoring results were recorded before the diet program and 2 and 4 weeks after the diet program. Acidic urinary pH was determined in 61.4% of patients with OAB. There was a significant association between the presence of acidic urine and OAB. Also, the OAB-V8 scores of patients were significantly higher in patients with acidic urine than nonacidic urine. OAB-V8 scores of patients showed statistically significant improvement after diet therapy (17.87 ± 6.52 vs. 10.43 ± 7.17; p < 0.001). We found that acidic urinary pH was closely associated with OAB, and alkalization of urine improved lower urinary tract symptoms. We suggest that urinary pH should be considered as a parameter in treatment planning of patients with OAB. © 2015 S. Karger AG, Basel.

  15. Na+-independent and nonstereospecific transport of 2-hydroxy 4-methylthiobutanoic acid by brush border membrane vesicles from chick small intestine.

    PubMed

    Brachet, P; Puigserver, A

    1989-01-01

    1. The transport of L- and DL-2-hydroxy 4-methylthiobutanoic acid (HMB), the methionine hydroxy analogue, by brush border membrane vesicles (BBMV) from chick small intestine was the sum of a saturable Michaelian component and a diffusive term. 2. Unlike that of L- and DL-MET, uptake was Na+-independent and electroneutral. 3. The inhibition of L-HMB transport by L-lactate, a structural analogue, and D-HMB as well, was of the competitive type. 4. Preloading of BBMV with D-HMB but not with L-lactate or L-MET trans-stimulated the influx of labelled L-HMB. 5. HMB uptake by rat and chick intestinal BBMV exhibited similar characteristics but the chick nonstereospecific transport system appeared to be unable to carry out L-lactate translocation.

  16. Implications of pH manipulation methods for metal toxicity: not all acidic environments are created equal.

    PubMed

    Esbaugh, A J; Mager, E M; Brix, K V; Santore, R; Grosell, M

    2013-04-15

    The toxicity of many metals is impacted by environmental pH, through both competition and complexation by hydroxide and carbonate ions. To establish safe environmental regulation it is important to properly define the relationship between pH and metal toxicity, a process that involves manipulating the pH of test water in the lab. The current study compares the effects of the three most common pH manipulation methods (carbon dioxide, acid-base addition, and chemical buffers) on acute Pb toxicity of a model fish species, Pimephales promelas. Acidification of test water revealed that the Pb and Pb(2+) LC50 values were impacted by the pH manipulation method, with the following order of effects: HClpH was alkalinized using MOPS or NaOH. The different impacts of pH manipulation methods on Pb toxicity are likely due to different physiological stresses resulting from the respective methods; the physiological implications of each method are discussed. The results suggest that when studying the impacts of pH on metal toxicity it is important to properly replicate the ambient conditions of interest as artificial buffering using CO2 environments or organic buffers significantly affects the physiology of the test organisms above and beyond what is expected from pH alone. Thus, using CO2 and organic buffers overestimates the impact of acid pH on Pb toxicity. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Basic principles of electrolyte chemistry for microfluidic electrokinetics. Part I: Acid-base equilibria and pH buffers.

    PubMed

    Persat, Alexandre; Chambers, Robert D; Santiago, Juan G

    2009-09-07

    We review fundamental and applied acid-base equilibrium chemistry useful to microfluidic electrokinetics. We present elements of acid-base equilibrium reactions and derive rules for pH calculation for simple buffers. We also present a general formulation to calculate pH of more complex, arbitrary mixtures of electrolytes, and discuss the effects of ionic strength and temperature on pH calculation. More practically, we offer advice on buffer preparation and on buffer reporting. We also discuss "real world" buffers and likely contamination sources. In particular, we discuss the effects of atmospheric carbon dioxide on buffer systems, namely, the increase in ionic strength and acidification of typical electrokinetic device buffers. In Part II of this two-paper series, we discuss the coupling of acid-base equilibria with electrolyte dynamics and electrochemistry in typical microfluidic electrokinetic systems.

  18. Using LysoSensor Yellow/Blue DND-160 to sense acidic pH under high hydrostatic pressures.

    PubMed

    DePedro, Hector Michael; Urayama, Paul

    2009-01-15

    LysoSensor Yellow/Blue DND-160, a dual-wavelength fluorophore commonly used for sensing pH in acidic organelles, possesses solvatochromic behavior believed to originate from an intramolecular charge transfer (ICT). Given this, we investigated whether DND-160 can be used for acidic pH sensing under hydrostatic pressures up to 510 atm, a range suitable for studying a wide variety of cellular processes. We found that the emission spectrum of the protonated form does not exhibit sensitivity to pressure, whereas the deprotonated form shows a piezochromic shift consistent with increased ICT character. Although pressure effects on the apparent pKa are buffer solvent dependent, DND-160 retains two-state behavior, making it a useful acidic pH probe under pressure.

  19. Development of On-Line Spectroscopic pH Monitoring for Nuclear Fuel Reprocessing Plants: Weak Acid Schemes

    SciTech Connect

    Casella, Amanda J.; Hylden, Laura R.; Campbell, Emily L.; Levitskaia, Tatiana G.; Peterson, James M.; Smith, Frances N.; Bryan, Samuel A.

    2015-05-19

    Knowledge of real-time solution properties and composition is a necessity for any spent nuclear fuel reprocessing method. Metal-ligand speciation in aqueous solutions derived from the dissolved commercial spent fuel is highly dependent upon the acid concentration/pH, which influences extraction efficiency and the resulting speciation in the organic phase. Spectroscopic process monitoring capabilities, incorporated in a counter current centrifugal contactor bank, provide a pathway for on-line real-time measurement of solution pH. The spectroscopic techniques are process-friendly and can be easily configured for on-line applications, while classic potentiometric pH measurements require frequent calibration/maintenance and have poor long-term stability in aggressive chemical and radiation environments. Our research is focused on developing a general method for on-line determination of pH of aqueous solutions through chemometric analysis of Raman spectra. Interpretive quantitative models have been developed and validated under the range of chemical composition and pH using a lactic acid/lactate buffer system. The developed model was applied to spectra obtained on-line during solvent extractions performed in a centrifugal contactor bank. The model predicted the pH within 11% for pH > 2, thus demonstrating that this technique could provide the capability of monitoring pH on-line in applications such as nuclear fuel reprocessing.

  20. [Characteristics of soil pH and exchangeable acidity in red soil profile under different vegetation types].

    PubMed

    Ji, Gang; Xu, Ming-gang; Wen, Shi-lin; Wang, Bo-ren; Zhang, Lu; Liu, Li-sheng

    2015-09-01

    The characteristics of soil pH and exchangeable acidity in soil profile under different vegetation types were studied in hilly red soil regions of southern Hunan Province, China. The soil samples from red soil profiles within 0-100 cm depth at fertilized plots and unfertilized plots were collected and analyzed to understand the profile distribution of soil pH and exchangeable acidity. The results showed that, pH in 0-60 cm soil from the fertilized plots decreased as the following sequence: citrus orchard > Arachis hypogaea field > tea garden. As for exchangeable acidity content, the sequence was A. hypogaea field ≤ citrus orchard < tea garden. After tea tree and A. hypogaea were planted for long time, acidification occurred in surface soil (0-40 cm), compared with the deep soil (60-100 cm), and soil pH decreased by 0.55 and 0.17 respectively, but such changes did not occur in citrus orchard. Soil pH in 0-40 cm soil from the natural recovery vegetation unfertilized plots decreased as the following sequence: Imperata cylindrica land > Castanea mollissima garden > Pinus elliottii forest ≥ Loropetalum chinensis forest. As for exchangeable acidity content, the sequence was L cylindrica land < C. mollissima garden < L. chinensis forest ≤ P. elliottii forest. Soil pH in surface soil (0-20 cm) from natural forest plots, secondary forest and Camellia oleifera forest were significantly lower than that from P. massoniana forest, decreased by 0.34 and 0.20 respectively. For exchangeable acidity content in 0-20 cm soil from natural forest plot, P. massoniana forest and secondary forest were significantly lower than C. oleifera forest. Compared with bare land, surface soil acidification in unfertilized plots except I. cylindrica land had been accelerated, and the natural secondary forest was the most serious among them, with surface soil pH decreasing by 0.52. However, the pH increased in deep soils from unfertilized plots except natural secondary forest, and I. cylindrica

  1. A cell-semiconductor synapse: transistor recording of vesicle release in chromaffin cells.

    PubMed

    Lichtenberger, Janosch; Fromherz, Peter

    2007-03-15

    The release of dense-core vesicles in bovine chromaffin cells is a model for the presynaptic process in neurons. It is usually studied by microamperometry of catecholamines with carbon fibers. Here we introduce transistor recording as a tool to study vesicle release. When we stimulate a chromaffin cell placed on a field-effect transistor, the gate voltage exhibits peaks that correlate with a simultaneously performed amperometric recording. We attribute the transistor signal to a release of protons from the extruded matrix of vesicles that lowers the extracellular pH and changes the electrical surface potential of the gate oxide. The rise time of the transistor signals is similar to that of amperometric responses, whereas their duration is distinctly longer. In a model computation, the rise time is identified with the extrusion of vesicle matrix into the narrow extracellular space between cell and gate oxide, and the decay time is attributed to pH equilibration through slow diffusion in the extruded matrix. Because the transistor recording relies on protons, it can be applied to acidic vesicles with electrochemically inactive hormones or transmitters.

  2. A Cell-Semiconductor Synapse: Transistor Recording of Vesicle Release in Chromaffin Cells

    PubMed Central

    Lichtenberger, Janosch; Fromherz, Peter

    2007-01-01

    The release of dense-core vesicles in bovine chromaffin cells is a model for the presynaptic process in neurons. It is usually studied by microamperometry of catecholamines with carbon fibers. Here we introduce transistor recording as a tool to study vesicle release. When we stimulate a chromaffin cell placed on a field-effect transistor, the gate voltage exhibits peaks that correlate with a simultaneously performed amperometric recording. We attribute the transistor signal to a release of protons from the extruded matrix of vesicles that lowers the extracellular pH and changes the electrical surface potential of the gate oxide. The rise time of the transistor signals is similar to that of amperometric responses, whereas their duration is distinctly longer. In a model computation, the rise time is identified with the extrusion of vesicle matrix into the narrow extracellular space between cell and gate oxide, and the decay time is attributed to pH equilibration through slow diffusion in the extruded matrix. Because the transistor recording relies on protons, it can be applied to acidic vesicles with electrochemically inactive hormones or transmitters. PMID:17189317

  3. On the Mechanism by which Alkaline pH Prevents Expression of an Acid-Expressed Gene

    PubMed Central

    Espeso, Eduardo A.; Arst, Herbert N.

    2000-01-01

    Previous work has shown that zinc finger transcription factor PacC mediates the regulation of gene expression by ambient pH in the fungus Aspergillus nidulans. This regulation ensures that the syntheses of molecules functioning in the external environment, such as permeases, secreted enzymes, and exported metabolites, are tailored to the pH of the growth environment. A direct role for PacC in activating the expression of an alkaline-expressed gene has previously been demonstrated, but the mechanism by which alkaline ambient pH prevents the expression of any eukaryotic acid-expressed gene has never been reported. Here we show that a double PacC binding site in the promoter of the acid-expressed gabA gene, encoding γ-aminobutyrate (GABA) permease, overlaps the binding site for the transcriptional activator IntA, which mediates ω-amino acid induction. Using bacterially expressed fusion proteins, we have shown that PacC competes with IntA for DNA binding in vitro at this site. Thus, PacC repression of GABA permease synthesis is direct and occurs by blocking induction. A swap of IntA sites between promoters for gabA and amdS, a gene not subject to pH regulation, makes gabA expression pH independent and amdS acid expressed. PMID:10779325

  4. Hydrothermal carbonization (HTC) of wheat straw: influence of feedwater pH prepared by acetic acid and potassium hydroxide.

    PubMed

    Reza, M Toufiq; Rottler, Erwin; Herklotz, Laureen; Wirth, Benjamin

    2015-04-01

    In this study, influence of feedwater pH (2-12) was studied for hydrothermal carbonization (HTC) of wheat straw at 200 and 260°C. Acetic acid and KOH were used as acidic and basic medium, respectively. Hydrochars were characterized by elemental and fiber analyses, SEM, surface area, pore volume and size, and ATR-FTIR, while HTC process liquids were analyzed by HPLC and GC. Both hydrochar and HTC process liquid qualities vary with feedwater pH. At acidic pH, cellulose and elemental carbon increase in hydrochar, while hemicellulose and pseudo-lignin decrease. Hydrochars produced at pH 2 feedwater has 2.7 times larger surface area than that produced at pH 12. It also has the largest pore volume (1.1 × 10(-1) ml g(-1)) and pore size (20.2 nm). Organic acids were increasing, while sugars were decreasing in case of basic feedwater, however, phenolic compounds were present only at 260°C and their concentrations were increasing in basic feedwater.

  5. Acidic pH and divalent cation sensing by PhoQ are dispensable for systemic salmonellae virulence.

    PubMed

    Hicks, Kevin G; Delbecq, Scott P; Sancho-Vaello, Enea; Blanc, Marie-Pierre; Dove, Katja K; Prost, Lynne R; Daley, Margaret E; Zeth, Kornelius; Klevit, Rachel E; Miller, Samuel I

    2015-05-23

    Salmonella PhoQ is a histidine kinase with a periplasmic sensor domain (PD) that promotes virulence by detecting the macrophage phagosome. PhoQ activity is repressed by divalent cations and induced in environments of acidic pH, limited divalent cations, and cationic antimicrobial peptides (CAMP). Previously, it was unclear which signals are sensed by salmonellae to promote PhoQ-mediated virulence. We defined conformational changes produced in the PhoQ PD on exposure to acidic pH that indicate structural flexibility is induced in α-helices 4 and 5, suggesting this region contributes to pH sensing. Therefore, we engineered a disulfide bond between W104C and A128C in the PhoQ PD that restrains conformational flexibility in α-helices 4 and 5. PhoQ(W104C-A128C) is responsive to CAMP, but is inhibited for activation by acidic pH and divalent cation limitation. phoQ(W104C-A128C) Salmonella enterica Typhimurium is virulent in mice, indicating that acidic pH and divalent cation sensing by PhoQ are dispensable for virulence.

  6. Metal reduction at low pH by a Desulfosporosinus species: implications for the biological treatment of acidic mine drainage

    SciTech Connect

    Senko, J.M.; Zhang, G.X.; McDonough, J.T.; Bruns, M.A.; Burgos, W.D.

    2009-07-01

    We isolated an acid-tolerant sulfate-reducing bacterium, GBSRB4.2, from coal mine-derived acidic mine drainage (AMD)-derived sediments. Sequence analysis of partial 16S rRNA gene of GBSRB4.2 revealed that it was affiliated with the genus Desulfosporosinus. GBSRB4.2 reduced sulfate, Fe(III) (hydr)oxide, Mn(IV) oxide, and U(VI) in acidic solutions (pH 4.2). Sulfate, Fe(III), and Mn(IV) but not U(VI) bioreduction led to an increase in the pH of acidic solutions and concurrent hydrolysis and precipitation of dissolved Al{sup 3+}. Reduction of Fe(III), Mn(IV), and U(VI) in sulfate-free solutions revealed that these metals are enzymatically reduced by GBSRB4.2. GBSRB4.2 reduced U(VI) in groundwater from a radionuclide-contaminated aquifer more rapidly at pH 4.4 than at pH 7.1, possibly due to the formation of poorly bioreducible Ca-U(VI)-CO{sub 3} complexes in the pH 7.1 groundwater.

  7. Acidic pH and divalent cation sensing by PhoQ are dispensable for systemic salmonellae virulence

    PubMed Central

    Hicks, Kevin G; Delbecq, Scott P; Sancho-Vaello, Enea; Blanc, Marie-Pierre; Dove, Katja K; Prost, Lynne R; Daley, Margaret E; Zeth, Kornelius; Klevit, Rachel E; Miller, Samuel I

    2015-01-01

    Salmonella PhoQ is a histidine kinase with a periplasmic sensor domain (PD) that promotes virulence by detecting the macrophage phagosome. PhoQ activity is repressed by divalent cations and induced in environments of acidic pH, limited divalent cations, and cationic antimicrobial peptides (CAMP). Previously, it was unclear which signals are sensed by salmonellae to promote PhoQ-mediated virulence. We defined conformational changes produced in the PhoQ PD on exposure to acidic pH that indicate structural flexibility is induced in α-helices 4 and 5, suggesting this region contributes to pH sensing. Therefore, we engineered a disulfide bond between W104C and A128C in the PhoQ PD that restrains conformational flexibility in α-helices 4 and 5. PhoQW104C-A128C is responsive to CAMP, but is inhibited for activation by acidic pH and divalent cation limitation. phoQW104C-A128C Salmonella enterica Typhimurium is virulent in mice, indicating that acidic pH and divalent cation sensing by PhoQ are dispensable for virulence. DOI: http://dx.doi.org/10.7554/eLife.06792.001 PMID:26002083

  8. Endogenous pH, Titratable Acidity and Total Soluble Solid Content of Mouthwashes Available in the Brazilian Market

    PubMed Central

    Cavalcanti, Alessandro Leite; Ramos, Ianny Alves; Leite, Rafaela Bastos; da Costa Oliveira, Mariana; de Melo Menezes, Karynna; Fernandes, Lígia Virgínio; de Castro, Ricardo Dias; Vieira, Fernando Fernandes

    2010-01-01

    Objectives: To evaluate in vitro the endogenous pH, titratable acidity and total soluble solid content (TSSC) of mouthwashes available in the Brazilian market. Methods: The study sample was composed of 10 commercial brands of mouthwashes based on different active ingredients: Cepacol®, Clinerize®, Equate®, Listerine Cool Citrus®, Oral-B®, Periogard®, Peroxyl®, Plax Overnight®, Prevident 220® and Sanifill®. The experiments were performed in triplicate. The endogenous pH was evaluated by potentiometry, titratable acidity was evaluated by the addition of 0.1N KOH increments to the mouthwashes, and TSSC readings were performed by Brix refractometry using the Abbé refractometer. Results: pH values ranged from 3.56 (Peroxyl®) to 7.43 (Cepacol®) and three mouthwashes presented pHs below 5.5. The titratable acidity values ranged from 0.007 (Periograd®) to 0.530 (Prevident®). Oral B® and Clinerize® presented the lowest (4.7%) and the highest (23.70%) TSSC, respectively. Conclusions: Some of the mouthwashes evaluated in this study presented low endogenous pH, even below the critical value for enamel dissolution (pH<5.5), high titratable acidity and high TSSC, and may be potentially erosive to the dental tissues if not properly used. PMID:20396446

  9. The pH profile for acid-induced elongation of coleoptile and epicotyl sections is consistent with the acid-growth theory

    NASA Technical Reports Server (NTRS)

    Cleland, R. E.; Buckley, G.; Nowbar, S.; Lew, N. M.; Stinemetz, C.; Evans, M. L.; Rayle, D. L.

    1991-01-01

    The acid-growth theory predicts that a solution with a pH identical to that of the apoplast of auxin-treated tissues (4.5.-5.0) should induce elongation at a rate comparable to that of auxin. Different pH profiles for elongation have been obtained, however, depending on the type of pretreatment between harvest of the sections and the start of the pH-incubations. To determine the acid sensitivity under in vivo conditions, oat (Avena sativa L.) coleoptile, maize (Zea mays L.) coleoptile and pea (Pisum sativum L.) epicotyl sections were abraded so that exogenous buffers could penetrate the free space, and placed in buffered solutions of pH 3.5-6.5 without any preincubation. The extension, without auxin, was measured over the first 3 h. Experiments conducted in three laboratories produced similar results. For all three species, sections placed in buffer without pretreatment elongated at least threefold faster at pH 5.0 than at 6.0 or 6.5, and the rate elongation at pH 5.0 was comparable to that induced by auxin. Pretreatment of abraded sections with pH-6.5 buffer or distilled water adjusted to pH 6.5 or above gave similar results. We conclude that the pH present in the apoplast of auxin-treated coleoptile and stems is sufficiently low to account for the initial growth response to auxin.

  10. The pH profile for acid-induced elongation of coleoptile and epicotyl sections is consistent with the acid-growth theory

    NASA Technical Reports Server (NTRS)

    Cleland, R. E.; Buckley, G.; Nowbar, S.; Lew, N. M.; Stinemetz, C.; Evans, M. L.; Rayle, D. L.

    1991-01-01

    The acid-growth theory predicts that a solution with a pH identical to that of the apoplast of auxin-treated tissues (4.5.-5.0) should induce elongation at a rate comparable to that of auxin. Different pH profiles for elongation have been obtained, however, depending on the type of pretreatment between harvest of the sections and the start of the pH-incubations. To determine the acid sensitivity under in vivo conditions, oat (Avena sativa L.) coleoptile, maize (Zea mays L.) coleoptile and pea (Pisum sativum L.) epicotyl sections were abraded so that exogenous buffers could penetrate the free space, and placed in buffered solutions of pH 3.5-6.5 without any preincubation. The extension, without auxin, was measured over the first 3 h. Experiments conducted in three laboratories produced similar results. For all three species, sections placed in buffer without pretreatment elongated at least threefold faster at pH 5.0 than at 6.0 or 6.5, and the rate elongation at pH 5.0 was comparable to that induced by auxin. Pretreatment of abraded sections with pH-6.5 buffer or distilled water adjusted to pH 6.5 or above gave similar results. We conclude that the pH present in the apoplast of auxin-treated coleoptile and stems is sufficiently low to account for the initial growth response to auxin.

  11. Acidic Nanoparticles Are Trafficked to Lysosomes and Restore an Acidic Lysosomal pH and Degradative Function to Compromised ARPE-19 Cells

    PubMed Central

    Baltazar, Gabriel C.; Guha, Sonia; Lu, Wennan; Lim, Jason; Boesze-Battaglia, Kathleen; Laties, Alan M.; Tyagi, Puneet; Kompella, Uday B.; Mitchell, Claire H.

    2012-01-01

    Lysosomal enzymes function optimally in acidic environments, and elevation of lysosomal pH can impede their ability to degrade material delivered to lysosomes through autophagy or phagocytosis. We hypothesize that abnormal lysosomal pH is a key aspect in diseases of accumulation and that restoring lysosomal pH will improve cell function. The propensity of nanoparticles to end up in the lysosome makes them an ideal method of delivering drugs to lysosomes. This study asked whether acidic nanoparticles could traffic to lysosomes, lower lysosomal pH and enhance lysosomal degradation by the cultured human retinal pigmented epithelial cell line ARPE-19. Acidic nanoparticles composed of poly (DL-lactide-co-glycolide) (PLGA) 502 H, PLGA 503 H and poly (DL-lactide) (PLA) colocalized to lysosomes of ARPE-19 cells within 60 min. PLGA 503 H and PLA lowered lysosomal pH in cells compromised by the alkalinizing agent chloroquine when measured 1 hr. after treatment, with acidification still observed 12 days later. PLA enhanced binding of Bodipy-pepstatin-A to the active site of cathepsin D in compromised cells. PLA also reduced the cellular levels of opsin and the lipofuscin-like autofluorescence associated with photoreceptor outer segments. These observations suggest the acidification produced by the nanoparticles was functionally effective. In summary, acid nanoparticles lead to a rapid and sustained lowering of lysosomal pH and improved degradative activity. PMID:23272048

  12. Effect of Acidic pH on Expression of Surface-Associated Proteins of Streptococcus oralis

    PubMed Central

    Wilkins, Joanna C.; Beighton, David; Homer, Karen A.

    2003-01-01

    Streptococcus oralis, a member of the mitis group of oral streptococci, is implicated in the pathogenesis of infective endocarditis and is the predominant aciduric non-mutans-group streptococcus in dental plaque. We undertook to identify the most abundant surface-associated proteins of S. oralis and to investigate changes in protein expression when the organism was grown under acidic culture conditions. Surface-associated proteins were extracted from cells grown in batch culture, separated by two-dimensional gel electrophoresis, excised, digested with trypsin, and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry and liquid chromatography-tandem mass spectrometry. Putative functions were assigned by homology to a translated genomic database of Streptococcus pneumoniae. A total of 27 proteins were identified; these included a lipoprotein, a ribosome recycling factor, and the glycolytic enzymes phosphoglycerate kinase, fructose bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, and enolase. The most abundant protein, phosphocarrier protein HPr, was present as three isoforms. Neither lactate dehydrogenase nor pyruvate oxidase, dominant intracellular proteins, were present among the proteins on the gels, demonstrating that proteins in the surface-associated pool did not arise as a result of cell lysis. Eleven of the proteins identified were differentially expressed when cells were grown at pH 5.2 versus pH 7.0, and these included superoxide dismutase, a homologue of dipeptidase V from Lactococcus lactis, and the protein translation elongation factors G, Tu, and Ts. This study has extended the range of streptococcal proteins known to be expressed at the cell surface. Further investigations are required to ascertain their functions at this extracellular location and determine how their expression is influenced by other environmental conditions. PMID:12957916

  13. Extracellular pH defense against lactic acid in untrained and trained altitude residents.

    PubMed

    Böning, D; Rojas, J; Serrato, M; Reyes, O; Coy, L; Mora, M

    2008-05-01

    The assumption that buffering at altitude is deteriorated by bicarbonate (bi) reduction was investigated. Extracellular pH defense against lactic acidosis was estimated from changes (Delta) in lactic acid ([La]), [HCO3-], pH and PCO2 in plasma, which equilibrates with interstitial fluid. These quantities were measured in earlobe blood during and after incremental bicycle exercise in 10 untrained (UT) and 11 endurance-trained (TR) highlanders (2,600 m). During exercise the capacity of non-bicarbonate buffers (betanbi=-Delta[La]. DeltapH(-1)-Delta[HCO3-]. DeltapH(-1)) amounted to 40+/-2 (SEM) and 28+/-2 mmol l(-1) in UT and TR, respectively (P<0.01). During recovery beta (nbi) decreased to 20 (UT) and 16 (TR) mmol l(-1) (P<0.001) corresponding to values expected from hemoglobin, dissolved protein and phosphate concentrations related to extracellular fluid (ecf). This was accompanied by a larger decrease of base excess after than during exercise for a given Delta[La]. betabi amounted to 37-41 mmol l(-1) being lower than at sea level. The large exercise betanbi was mainly caused by increasing concentrations of buffers due to temporary shrinking of ecf. Tr has lower betanbi in spite of an increased Hb mass mainly because of an expanded ecf compared to UT. In highlanders betanbi is higher than in lowlanders because of larger Hb mass and reduced ecf and counteracts the decrease in [HCO3-]. The amount of bicarbonate is probably reduced by reduction of the ecf at altitude but this is compensated by lower maximal [La] and more effective hyperventilation resulting in attenuated exercise acidosis at exhaustion.

  14. Effect of temperature on pH measurements and acid-base equilibria in methanol-water mixtures.

    PubMed

    Castells, Cecilia B; Ràfols, Clara; Rosés, Martí; Bosch, Elisabeth

    2003-06-20

    The knowledge of the acid-base equilibria in water-solvent mixtures of both common buffers and analytes is necessary in order to predict their retention as function of pH, solvent composition and temperature. This paper describes the effect of temperature on acid-base equilibria in methanol-water solvent mixtures commonly used as HPLC mobile phases. We measured the delta-correction parameter (delta = sw pH - ss pH = Ej - log sw(gamma)oh) between two pH scales: pH measured in the solvent concerned and referred to the same standard state, ss pH, and the pH measured in that solvent mixture but referred to water as standard state, sw pH, for several methanol compositions in the temperature range of 20-50 degrees C. These determinations suggest that the delta-term depends only on composition of the mixture and on temperature. In water-rich mixtures, for which methanol is below 40% (w/w), delta-term seems to be independent of temperature, within the experimental uncertainties, whereas for methanol content larger than 50% (w/w) the delta-correction decreases as temperature increases. We have attributed this decrease to a large increase in the medium effect when mixtures have more than 50% methanol. The pKa of five weak electrolytes of different chemical nature in 50% methanol-water at 20-50 degrees C are presented: the effect of temperature on pKa was large for amines, pyridine and phenol, but almost no dependence was found for benzoic acid. This indicates that buffers can play a critical role in affecting retention and selectivity in HPLC at temperatures far from 25 degrees C, particularlyfor co-eluted solutes.

  15. Buffers for the physiological pH range: acidic dissociation constants of zwitterionic compounds in various hydroorganic media.

    PubMed

    Taha, Mohamed

    2005-01-01

    The measurement of pH of a physiological buffer is an integral part of clinical diagnosis. For the standardization of pH and control of acidity in the physiological region of pH 7 to 9, as found in blood and plasma. Good et al., Ferguson et al., and Bellini et al. have listed hydrogen buffers which are N-substituted amino acids compatible with common biological media. Recently, the acid base behavior of BICINE, in aqueous solution and in different solvent mixtures has been studied by us. Other investigators reported similar results for (CAPS and CAPSO), (BES), (ACES and CHES), TRICINE, TES, ADA, (HEPPS and MOPSO), (MOPS), and (MOBS and HEPBS). These studies have provided precise knowledge about the suitability of a biological buffer. No work seems to have been done on the determination of second stage dissociation constants of 2-(4-Morphlinomethyl)propenoic acid, 2-[bis(2-hydroxyethyl)amino methyl]propenoic acid, 2-[bis(2-hydroxypropyl)aminomethyl]propenoic acid, and 2-[N-ethyl-N-(2-hydroxyethyl)aminomethyl]propenoic acid in aquo-organic solvent mixtures.

  16. Growth and Metabolism of Lactic Acid Bacteria during and after Malolactic Fermentation of Wines at Different pH

    PubMed Central

    Davis, C. R.; Wibowo, D. J.; Lee, T. H.; Fleet, G. H.

    1986-01-01

    Commercially produced red wines were adjusted to pH 3.0, 3.2, 3.5, 3.7, or 4.0 and examined during and after malolactic fermentation for growth of lactic acid bacteria and changes in the concentrations of carbohydrates, organic acids, amino acids, and acetaldehyde. With one exception, Leuconostoc oenos conducted the malolactic fermentation in all wines and was the only species to occur in wines at pH below 3.5. Malolactic fermentation by L. oenos was accompanied by degradation of malic, citric, and fumaric acids and production of lactic and acetic acids. The concentrations of arginine, histidine, and acetaldehyde also decreased at this stage, but the behavior of hexose and pentose sugars was complicated by other factors. Pediococcus parvulus conducted the malolactic fermentation in one wine containing 72 mg of total sulfur dioxide per liter. Fumaric and citric acids were not degraded during this malolactic fermentation, but hexose sugars were metabolized. P. parvulus and species of Lactobacillus grew after malolactic fermentation in wines with pH adjusted above 3.5. This growth was accompanied by the utilization of wine sugars and production of lactic and acetic acids. PMID:16347015

  17. A theoretical study on the pH dependence of X-ray emission spectra for aqueous acetic acid

    NASA Astrophysics Data System (ADS)

    Nishida, Naohiro; Tokushima, Takashi; Takahashi, Osamu

    2016-04-01

    We performed theoretical calculations to reproduce the site-selective XES spectra for aqueous acetic acid at the oxygen K-edge. The shape of the experimental XES spectra obtained from aqueous acetic acid drastically changed when the pH value was high. Structure sampling of an aqueous acetic acid cluster model was performed by the ab initio molecular dynamics trajectory. Relative XES peak intensities for the core⿿hole excited state dynamics simulations were calculated using density functional theory. We found that the theoretical XES spectra reproduced well the experimental spectra and that these calculations gave us electronic and molecular structure information about aqueous acetic acid.

  18. A pH and thermosensitive choline phosphate-based delivery platform targeted to the acidic tumor microenvironment.

    PubMed

    Yu, Xifei; Yang, Xiaoqiang; Horte, Sonja; Kizhakkedathu, Jayachandran N; Brooks, Donald E

    2014-01-01

    Solid tumors generally exhibit an acidic microenvironment which has been recognized as a potential route to distinguishing tumor from normal tissue for purposes of drug delivery or imaging. To this end we describe a pH and temperature sensitive polymeric adhesive that can be derivatized to carry drugs or other agents and can be tuned synthetically to bind to tumor cells at pH 6.8 but not at pH 7.4 at 37 °C. The adhesive is based on the universal reaction between membrane phosphatidyl choline (PC) molecules and polymers derivatized with multiple copies of the inverse motif, choline phosphate (CP). The polymer family we use is a linear copolymer of a CP terminated tetraethoxymethacrylate and dimethylaminoethyl (DMAE) methacrylate, the latter providing pH sensitivity. The copolymer exhibits a lower critical solution temperature (LCST) just below 37 °C when the DMAE is uncharged at pH 7.4 but the LCST does not occur when the group is charged at pH 6.8 due to the ionization hydrophilicity. At 37 °C the polymer binds strongly to mammalian cells at pH 6.8 but does not bind at pH 7.4, potentially targeting tumor cells existing in an acidic microenvironment. We show the binding is strong, reversible if the pH is raised and is followed rapidly by cellular uptake of the fluorescently labeled material. Drug delivery utilizing this dually responsive family of polymers should provide a basis for targeting tumor cells with minimal side reactions against untransformed counterparts.

  19. Evaluation of acidity and total sugar content of children's popular beverages and their effect on plaque pH.

    PubMed

    Saeed, S; Al-Tinawi, M

    2010-01-01

    In the developing countries, dental caries has increased with the increased exposure to dietary sugars. There is no data on the cariogenicity and acidogenicity of popular beverages in the Syrian market. To investigate the endogenous pH, titratable acidity, and total sugar content of popular beverages (cola, orange juice, and full-fat milk), and assess plaque pH drop after consumption. Twenty-five healthy children with a mean age of 11.8 ± 0.6 years were recruited for this single blinded study. The pH of beverages was measured by a digital pH meter and the titratable acidity was expressed as the volume of 0.1 N sodium hydroxide required to neutralize the beverage. The total sugar content was estimated using High Performance Liquid Chromatography. Children rinsed with 15 mL of each of the beverages for 1 min. The controls used were 10% sucrose and 10% sorbitol solutions. The plaque pH was assessed before and after rinsing. Statistical analysis was performed using analysis of variance followed by Bonferroni test to assess minimum pH, maximum pH drop, and the area under curve. P value was set as 0.05. Both cola and orange juice had a low pH and similar total sugar content in contrast to the high pH and low sugar content of milk. Cola and orange juice were not statistically different from 10% sucrose (P > 0.05), but different from 10% sorbitol (P < 0.05), in contrast to milk, for all the parameters studied. Cola and orange juice are cariogenic/acidogenic and frequent intake should be discouraged. A reasonable intake of unsweetened milk may be advised safely.

  20. Nonenzymatic glycation of phosphatidylethanolamine in erythrocyte vesicles

    SciTech Connect

    Patkowska, M.J.; Horowitz, M.I.

    1986-05-01

    Unsealed inside-out and right-side out vesicles were prepared from human red cells. The vesicles were incubated with D-glucose (/sup 14/C(U)) and sodium cyanoborohydride in phosphate buffer, pH 7.4. After incubation, lipids were extracted with 1-butanol and non-lipid contaminants removed by Sephadex G-25 chromatography. Phosphatidylethanolamine-sorbitol was purified by chromatography on columns of silicic acid and phenylboronate agarose gel. Phospholipase C (B. cereus) liberated phosphoethanolamine-sorbitol (I) which comigrated on TLC with synthetic I prepared by reductive condensation of phosphoethanolamine and D-glucose and also with the product of phospholipase C (B. cereus) hydrolysis of reference phosphatidylethanolamine-sorbitol. Exposure of I to alkaline phosphatase (E. coli) gave P/sub i/ and ethanolamine-sorbitol (II) which comigrated on TLC with synthetic II prepared by reductive condensation of ethanolamine and D-glucose or by phospholipase D hydrolysis of reference phosphatidylethanolamine-sorbitol. These studies demonstrate that vesicular phosphatidylethanolamine can be reductively glycated and illustrate the applicability of both phospholipase C and phospholipase D in characterizing glycated phosphoglycerides.

  1. Effects of acetic acid and arginine on pH elevation and growth of Bacillus licheniformis in an acidified cucumber juice medium.

    PubMed

    Yang, Zhenquan; Meng, Xia; Breidt, Frederick; Dean, Lisa L; Arritt, Fletcher M

    2015-04-01

    Bacillus licheniformis has been shown to cause pH elevation in tomato products having an initial pH below 4.6 and metabiotic effects that can lead to the growth of pathogenic bacteria. Because of this, the organism poses a potential risk to acidified vegetable products; however, little is known about the growth and metabolism of this organism in these products. To clarify the mechanisms of pH change and growth of B. licheniformis in vegetable broth under acidic conditions, a cucumber juice medium representative of a noninhibitory vegetable broth was used to monitor changes in pH, cell growth, and catabolism of sugars and amino acids. For initial pH values between pH 4.1 to 6.0, pH changes resulted from both fermentation of sugar (lowering pH) and ammonia production (raising pH). An initial pH elevation occurred, with starting pH values of pH 4.1 to 4.9 under both aerobic and anaerobic conditions, and was apparently mediated by the arginine deiminase reaction of B. licheniformis. This initial pH elevation was prevented if 5 mM or greater acetic acid was present in the brine at the same pH. In laboratory media, under favorable conditions for growth, data indicated that growth of the organism was inhibited at pH 4.6 with protonated acetic acid concentrations of 10 to 20 mM, corresponding to 25 to 50 mM total acetic acid; however, growth inhibition required greater than 300 mM citric acid (10-fold excess of the amount in processed tomato products) products under similar conditions. The data indicate that growth and pH increase by B. licheniformis may be inhibited by the acetic acid present in most commercial acidified vegetable products but not by the citric acid in many tomato products.

  2. Effect of acidity consumption/production on the pH of aeration tank during the biodegradation of acetic acid/epichlorohydrin.

    PubMed

    Yoon, Seong-Hoon; Kim, Hyung-Soo; Chung, Yun-Chul

    2002-06-01

    In order to elucidate the biologically driven pH fluctuation phenomena in industrial wastewater treatment, the contrary effects of acetic acid (AA) and epichlorohydrin (ECH) on the pH of aeration tank were investigated. Two simple equations were derived to estimate optimum neutralization pHs for the biological AA/ECH wastewater treatment, and the calculated optimum neutralization pHs were compared with experimental results. The pH in aeration tank was expected to fluctuate sharply with the smallest deviation of neutralization pH from the optimum value. However experimental results showed that real pH fluctuation is smaller than the theoretical one. It was considered that carbonate buffer in aqueous system relieves the pH fluctuation. The deviation between experimental and theoretical optimum neutralization pH could be mainly caused by volatility of AA and ECH. The deviation was larger with ECH wastewater of which volatility is larger than AA. Finally, this theory was successfully applied to the real petrochemical wastewater treatment. The pH of aeration tank was properly maintained when acidified wastewater (pH 3.4) was supplied.

  3. Model cell membranes: Techniques to form complex biomimetic supported lipid bilayers via vesicle fusion

    PubMed Central

    Hardy, Gregory J.; Nayak, Rahul

    2013-01-01

    Vesicle fusion has long provided an easy and reliable method to form supported lipid bilayers (SLBs) from simple, zwitterionic vesicles on siliceous substrates. However, for complex compositions, such as vesicles with high cholesterol content and multiple lipid types, the energy barrier for the vesicle-to-bilayer transition is increased or the required vesicle-vesicle and vesicle-substrate interactions are insufficient for vesicle fusion. Thus, for vesicle compositions that more accurately mimic native membranes, vesicle fusion often fails to form SLBs. In this paper, we review three approaches to overcome these barriers to form complex, biomimetic SLBs via vesicle fusion: (i) optimization of experimental conditions (e.g., temperature, buffer ionic strength, osmotic stress, cation valency, and buffer pH), (ii) α-helical (AH) peptide-induced vesicle fusion, and (iii) bilayer edge-induced vesicle fusion. AH peptide-induced vesicle fusion can form complex SLBs on multiple substrate types without the use of additional equipment. Bilayer edge-induced vesicle fusion uses microfluidics to form SLBs from vesicles with complex composition, including vesicles derived from native cell membranes. Collectively, this review introduces vesicle fusion techniques that can be generalized for many biomimetic vesicle compositions and many substrate types, and thus will aid efforts to reliably create complex SLB platforms on a range of substrates. PMID:24031164

  4. Uptake of auxins into membrane vesicles isolated from pea stems: an in vitro auxin transport system

    SciTech Connect

    Slone, J.H.

    1985-01-01

    The objective of this research was to test the applicability of the chemiosmotic theory of auxin transport to a subcellular system. Membrane vesicles were isolated from the basal portion of the third internode of etiolated pea plants (Pisum sativum L. var. Alaska) by differential centrifugation. Uptake of auxin was determined by adding /sup 14/C-labeled indoleacetic acid (IAA) to vesicles. Nigericin, a monovalent cation ionophore, and the electrogenic protonophore, carbonyl-cyanide m-chlorophenylhydrazone (CCCP), at micromolar concentrations abolished saturable uptake. Bursting vesicles by sonication, osmotic shock and freeze/thawing also eliminated saturable uptake. As the temperature increased from 0 to 30/sup 0/C, saturable uptake decreased markedly. Nonsaturable auxin uptake was less affected by these treatments. The pH gradient-dependent uptake of auxin appeared to be a transmembrane uptake of auxin into the vesicles rather than surface binding. Unlabeled IAA, 2,4-dichlorophenoxyacetic acid (2,4-D) and 2-naphthaleneacetic acid (NAA) at low concentrations reduced the saturable accumulation of (/sup 14/C)IAA in vesicles, while phenylacetic acid, benzoic acid, and 1-NAA were effective only at high concentrations. Kinetic analysis revealed two types of sites: a high affinity site with an uptake capacity of 25 to 40 pmoles/g tissue, and a low affinity site with an uptake capacity of 260 to 600 pmole/g tissue, fresh wt. In conclusion, several principal elements of an auxin transport system, as specific by the chemiosmotic theory of polar auxin transport, were present in membrane vesicles isolated from relatively mature pea stem tissue. However, one important aspect of the theory was not demonstrated in this in vitro system - a TIBA/NPA-sensitive auxin efflux. The kinetics and specificity of auxin uptake strongly suggested that this system was physiologically significant.

  5. Sodium dodecyl sulfate/β-cyclodextrin vesicles embedded in chitosan gel for insulin delivery with pH-selective release.

    PubMed

    Li, Zhuo; Li, Haiyan; Wang, Caifen; Xu, Jianghui; Singh, Vikramjeet; Chen, Dawei; Zhang, Jiwen

    2016-07-01

    In an answer to the challenge of enzymatic instability and low oral bioavailability of proteins/peptides, a new type of drug-delivery vesicle has been developed. The preparation, based on sodium dodecyl sulfate (SDS) and β-cyclodextrin (β-CD) embedded in chitosan gel, was used to successfully deliver the model drug-insulin. The self-assembled SDS/β-CD vesicles were prepared and characterized by particle size, zeta potential, appearance, microscopic morphology and entrapment efficiency. In addition, both the interaction of insulin with vesicles and the stability of insulin loaded in vesicles in the presence of pepsin were investigated. The vesicles were crosslinked into thermo-sensitive chitosan/β-glycerol phosphate solution for an in-situ gel to enhance the dilution stability. The in vitro release characteristics of insulin from gels in media at different pH values were investigated. The insulin loaded vesicles-chitosan hydrogel (IVG) improved the dilution stability of the vesicles and provided pH-selective sustained release compared with insulin solution-chitosan hydrogel (ISG). In vitro, IVG exhibited slow release in acidic solution and relatively quick release in neutral solutions to provide drug efficacy. In simulated digestive fluid, IVG showed better sustained release and insulin protection properties compared with ISG. Thus IVG might improve the stability of insulin during its transport in vivo and contribute to the bioavailability and therapeutic effect of insulin.

  6. Adsorption of organic acids on TiO2 nanoparticles: effects of pH, nanoparticle size, and nanoparticle aggregation.

    PubMed

    Pettibone, John M; Cwiertny, David M; Scherer, Michelle; Grassian, Vicki H

    2008-06-01

    In this study, the adsorption of two organic acids, oxalic acid and adipic acid, on TiO2 nanoparticles was investigated at room temperature, 298 K. Solution-phase measurements were used to quantify the extent and reversibility of oxalic acid and adipic acid adsorption on anatase nanoparticles with primary particle sizes of 5 and 32 nm. At all pH values considered, there were minimal differences in measured Langmuir adsorption constants, K ads, or surface-area-normalized maximum adsorbate-surface coverages, Gamma max, between 5 and 32 nm particles. Although macroscopic differences in the reactivity of these organic acids as a function of nanoparticle size were not observed, ATR-FTIR spectroscopy showed some distinct differences in the absorption bands present for oxalic acid adsorbed on 5 nm particles compared to 32 nm particles, suggesting different adsorption sites or a different distribution of adsorption sites for oxalic acid on the 5 nm particles. These results illustrate that molecular-level differences in nanoparticle reactivity can still exist even when macroscopic differences are not observed from solution phase measurements. Our results also allowed the impact of nanoparticle aggregation on acid uptake to be assessed. It is clear that particle aggregation occurs at all pH values and that organic acids can destabilize nanoparticle suspensions. Furthermore, 5 nm particles can form larger aggregates compared to 32 nm particles under the same conditions of pH and solid concentrations. The relative reactivity of 5 and 32 nm particles as determined from Langmuir adsorption parameters did not appear to vary greatly despite differences that occur in nanoparticle aggregation for these two different size nanoparticles. Although this potentially suggests that aggregation does not impact organic acid uptake on anatase particles, these data clearly show that challenges remain in assessing the available surface area for adsorption in nanoparticle aqueous suspensions

  7. Gallic Acid as a Complexing Agent for Copper Chemical Mechanical Polishing Slurries at Neutral pH

    NASA Astrophysics Data System (ADS)

    Kim, Yung Jun; Kang, Min Cheol; Kwon, Oh Joong; Kim, Jae Jeong

    2011-05-01

    Gallic acid was investigated as a new complexing agent for copper (Cu) chemical mechanical polishing slurries at neutral pH. Addition of 0.03 M gallic acid and 1.12 M H2O2 at pH 7 resulted in a Cu removal rate of 560.73±17.49 nm/min, and the ratio of the Cu removal rate to the Cu dissolution rate was 14.8. Addition of gallic acid improved the slurry performance compared to glycine addition. X-ray photoelectron spectroscopy analysis and contact angle measurements showed that addition of gallic acid enhanced the Cu polishing behavior by suppressing the formation of surface Cu oxide.

  8. Chemical crosslinking of acrylic acid to form biocompatible pH sensitive hydrogel reinforced with cellulose nanocrystals (CNC)

    NASA Astrophysics Data System (ADS)

    Lim, Lim Sze; Ahmad, Ishak; Lazim, Mohd Azwani Shah Mat; Amin, Mohd. Cairul Iqbal Mohd

    2014-09-01

    The purpose of this study is to produce a novel pH and temperature sensitive hydrogel, composed of poly(acrylic acid) (PAA) and cellulose nanocrystal (CNC). CNC was extracted from kenaf fiber through a series of alkali and bleaching treatments followed by acid hydrolysis. The PAA was then subjected to chemical cross-linking using the cross-linking agent (N,N-methylenebisacrylamide) with CNC entrapped in PAA matrix. The mixture was casted onto petri dish to obtain disc shape hydrogel. The effects of reaction conditions such as the ratio of PAA and CNC on the swelling behavior of the hydrogel obtained towards pH and temperature were studied. The obtained hydrogel was further subjected to different tests such swelling test for swelling behaviour at different pH and temperature along with scanning electron microscopy (SEM) for morphology analysis. The hydrogel obtained showed excellent pH sensitivity and obtained maximum swelling at pH 7. Besides that, hydrogel obtained showed significant increase in swelling ratio when temperature of swelling medium was increased from 25°C to 37°C. SEM micrograph showed that the pore size of the hydrogel decreases with increase of CNC content proving that the hydrogel structure became more rigid with addition of CNC. The PAA/CNC hydrogel with such excellent sensitivity towards pH and temperature can be developed further as drug carrier.

  9. Chemical crosslinking of acrylic acid to form biocompatible pH sensitive hydrogel reinforced with cellulose nanocrystals (CNC)

    SciTech Connect

    Lim, Lim Sze; Ahmad, Ishak; Lazim, Mohd Azwani Shah Mat; Amin, Mohd. Cairul Iqbal Mohd

    2014-09-03

    The purpose of this study is to produce a novel pH and temperature sensitive hydrogel, composed of poly(acrylic acid) (PAA) and cellulose nanocrystal (CNC). CNC was extracted from kenaf fiber through a series of alkali and bleaching treatments followed by acid hydrolysis. The PAA was then subjected to chemical cross-linking using the cross-linking agent (N,N-methylenebisacrylamide) with CNC entrapped in PAA matrix. The mixture was casted onto petri dish to obtain disc shape hydrogel. The effects of reaction conditions such as the ratio of PAA and CNC on the swelling behavior of the hydrogel obtained towards pH and temperature were studied. The obtained hydrogel was further subjected to different tests such swelling test for swelling behaviour at different pH and temperature along with scanning electron microscopy (SEM) for morphology analysis. The hydrogel obtained showed excellent pH sensitivity and obtained maximum swelling at pH 7. Besides that, hydrogel obtained showed significant increase in swelling ratio when temperature of swelling medium was increased from 25°C to 37°C. SEM micrograph showed that the pore size of the hydrogel decreases with increase of CNC content proving that the hydrogel structure became more rigid with addition of CNC. The PAA/CNC hydrogel with such excellent sensitivity towards pH and temperature can be developed further as drug carrier.

  10. Influence of acidic pH on the formulation of TiO2 nanocrystalline powders with enhanced photoluminescence property.

    PubMed

    Tsega, Moges; Dejene, F B

    2017-02-01

    Titanium dioxide (TiO2) nanoparticles were prepared by the sol-gel method at different pH values (3.2-6.8) with a hydrochloric acid (HCl) solution. Raw samples were calcined at 500 °C for 2 h. The effects of pH on the structural, morphological and optical properties of TiO2 nanoparticles were investigated. At pH 4.4-6.8, only the anatase phase of TiO2 was observed. Under strong acidic condition at pH 3.2 rutile, brookite and anatase co-exist, but rutile is the predominant phase. The strain value increased and the crystallite size decreased as the HCl content increased. The increased crystallite sizes in the range 21-24 nm and enhanced blue emission intensity around 432 nm was obtained for the sample at pH 5.0. Experimental results showed that TiO2 nanoparticles synthesized at pH 5.0 exhibited the best luminescence property with pure anatase phase.

  11. The Potential Role of Systemic Buffers in Reducing Intratumoral Extracellular pH and Acid-Mediated Invasion

    PubMed Central

    Silva, Ariosto S.; Yunes, Jose A.; Gillies, Robert J.; Gatenby, Robert A.

    2013-01-01

    A number of studies have shown that the extracellular pH (pHe) in cancers is typically lower than that in normal tissue and that an acidic pHe promotes invasive tumor growth in primary and metastatic cancers. Here, we investigate the hypothesis that increased systemic concentrations of pH buffers reduce intratumoral and peritumoral acidosis and, as a result, inhibit malignant growth. Computer simulations are used to quantify the ability of systemic pH buffers to increase the acidic pHe of tumors in vivo and investigate the chemical specifications of an optimal buffer for such purpose. We show that increased serum concentrations of the sodium bicarbonate (NaHCO3) can be achieved by ingesting amounts that have been used in published clinical trials. Furthermore, we find that consequent reduction of tumor acid concentrations significantly reduces tumor growth and invasion without altering the pH of blood or normal tissues. The simulations also show that the critical parameter governing buffer effectiveness is its pKa. This indicates that NaHCO3, with a pKa of 6.1, is not an ideal intratumoral buffer and that greater intratumoral pHe changes could be obtained using a buffer with a pKa of ~7. The simulations support the hypothesis that systemic pH buffers can be used to increase the tumor pHe and inhibit tumor invasion. PMID:19276380

  12. The potential role of systemic buffers in reducing intratumoral extracellular pH and acid-mediated invasion.

    PubMed

    Silva, Ariosto S; Yunes, Jose A; Gillies, Robert J; Gatenby, Robert A

    2009-03-15

    A number of studies have shown that the extracellular pH (pHe) in cancers is typically lower than that in normal tissue and that an acidic pHe promotes invasive tumor growth in primary and metastatic cancers. Here, we investigate the hypothesis that increased systemic concentrations of pH buffers reduce intratumoral and peritumoral acidosis and, as a result, inhibit malignant growth. Computer simulations are used to quantify the ability of systemic pH buffers to increase the acidic pHe of tumors in vivo and investigate the chemical specifications of an optimal buffer for such purpose. We show that increased serum concentrations of the sodium bicarbonate (NaHCO(3)) can be achieved by ingesting amounts that have been used in published clinical trials. Furthermore, we find that consequent reduction of tumor acid concentrations significantly reduces tumor growth and invasion without altering the pH of blood or normal tissues. The simulations also show that the critical parameter governing buffer effectiveness is its pK(a). This indicates that NaHCO(3), with a pK(a) of 6.1, is not an ideal intratumoral buffer and that greater intratumoral pHe changes could be obtained using a buffer with a pK(a) of approximately 7. The simulations support the hypothesis that systemic pH buffers can be used to increase the tumor pHe and inhibit tumor invasion.

  13. Effect of urinary pH on the pharmacokinetics of salicylic acid, with its glycine and glucuronide conjugates in human.

    PubMed

    Vree, T B; Van Ewijk-Beneken Kolmer, E W; Verwey-Van Wissen, C P; Hekster, Y A

    1994-10-01

    We studied the effects of urinary pH on the kinetics of salicylic acid (SA) with its metabolites and assessed the contribution of alkaline hydrolysis of salicylic acid acyl glucuronide to the renal clearance of salicylic acid. Hydrolysis of SAAG in alkaline urine contributes marginally to the high renal clearance and excretion of salicylic acid, validating alkalinization of a patient with SA overdose. Under acidic urine conditions, salicylic acid (SA) had a terminal plasma t1/2 value of 3.29 +/- 0.52 hours while under alkaline urine conditions this t1/2 was significantly reduced to 2.50 +/- 0.41 hours (p = 0.0156). The total oral body clearance of salicylic acid under acidic conditions (1.38 +/- 0.43 l/h) is significantly lower than under alkaline urine conditions (2.27 +/- 0.83 l/h; p = 0.0410). The Km and Vmax values of SA, and its conjugates salicylic acid phenolic glucuronide (SAPG), salicyluric acid (SU) and salicyluric acid phenolic glucuronide (SUPG) did not differ statistically under acidic and alkaline urine conditions. The protein binding of SA was 93.8 +/- 1.0% and that of SU was 89.7 +/- 2.2% in vivo and in vitro. SUPG had a protein binding of 84.8 +/- 1.8%, while SAPG showed no protein binding at all. The renal excretion of salicylic acid depends strongly on the urinary pH. The percentage of the dose excreted unchanged increased from 2.3 +/- 1.5% under acidic conditions to 30.5 +/- 9.1% under alkaline conditions (p = 0.0006). Alkaline urine lowered by 50% the percentage of the dose excreted as SU (p = 0.0028), SAAG (p = 0.0013), and SUPG (p = 0.0296), while SAPG is only marginally lowered (p = 0.0589).(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Fast hydrazone reactants: electronic and acid/base effects strongly influence rate at biological pH.

    PubMed

    Kool, Eric T; Park, Do-Hyoung; Crisalli, Pete

    2013-11-27

    Kinetics studies with structurally varied aldehydes and ketones in aqueous buffer at pH 7.4 reveal that carbonyl compounds with neighboring acid/base groups form hydrazones at accelerated rates. Similarly, tests of a hydrazine with a neighboring carboxylic acid group show that it also reacts at an accelerated rate. Rate constants for the fastest carbonyl/hydrazine combinations are 2-20 M(-1) s(-1), which is faster than recent strain-promoted cycloaddition reactions.

  15. Determination of critical pH and Al concentration of acidic Ultisols for wheat and canola crops

    NASA Astrophysics Data System (ADS)

    Abdulaha-Al Baquy, M.; Li, Jiu-Yu; Xu, Chen-Yang; Mehmood, Khalid; Xu, Ren-Kou

    2017-02-01

    Soil acidity has become a principal constraint in dry land crop production systems of acidic Ultisols in tropical and subtropical regions of southern China, where winter wheat and canola are cultivated as important rotational crops. There is little information on the determination of critical soil pH as well as aluminium (Al) concentration for wheat and canola crops. The objective of this study is to determine the critical soil pH and exchangeable aluminium concentration (AlKCl) for wheat and canola production. Two pot cultures with two Ultisols from Hunan and Anhui (SE China) were conducted for wheat and canola crops in a controlled growth chamber. Aluminium sulfate (Al2(SO4)3) and hydrated lime (Ca(OH)2) were used to obtain the target soil pH levels from 3.7 (Hunan) and 3.97 (Anhui) to 6.5. Plant height, shoot dry weight, root dry weight, and chlorophyll content (SPAD value) of wheat and canola were adversely affected by soil acidity in both locations. The critical soil pH and AlKCl of the Ultisol from Hunan for wheat were 5.29 and 0.56 cmol kg-1, respectively. At Anhui, the threshold soil pH and AlKCl for wheat were 4.66 and 1.72 cmol kg-1, respectively. On the other hand, the critical soil pH for canola was 5.65 and 4.87 for the Ultisols from Hunan and Anhui, respectively. The critical soil exchangeable Al for canola cannot be determined from the experiment of this study. The results suggested that the critical soil pH and AlKCl varied between different locations for the same variety of crop, due to the different soil types and their other soil chemical properties. The critical soil pH for canola was higher than that for wheat for both Ultisols, and thus canola was more sensitive to soil acidity. Therefore, we recommend that liming should be undertaken to increase soil pH if it falls below these critical soil pH levels for wheat and canola production.

  16. Charge-transfer-based terbium MOF nanoparticles as fluorescent pH sensor for extreme acidity.

    PubMed

    Qi, Zewan; Chen, Yang

    2017-01-15

    Newly emerged metal organic frameworks (MOFs) have aroused the great interest in designing functional materials by means of its flexible structure and component. In this study, we used lanthanide Tb(3+) ions and small molecular ligands to design and assemble a kind of pH-sensitive MOF nanoparticle based on intramolecular-charge-transfer effect. This kind of made-to-order MOF nanoparticle for H(+) is highly specific and sensitive and could be used to fluorescently indicate pH value of strong acidic solution via preset mechanism through luminescence of Tb(3+). The long luminescence lifetime of Tb(3+) allows eliminating concomitant non-specific fluorescence by time-revised fluorescence techniques, processing an advantage in sensing H(+) in biological media with strong autofluorescence. Our method showed a great potential of MOF structures in designing and constructing sensitive sensing materials for specific analytes directly via the assembly of functional ions/ligands. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Biophysical implications of sphingosine accumulation in membrane properties at neutral and acidic pH.

    PubMed

    Zupancic, Eva; Carreira, Ana C; de Almeida, Rodrigo F M; Silva, Liana C

    2014-05-08

    Sphingosine (Sph) is a simple lipid involved in the regulation of several biological processes. When accumulated in the late endosomal/lysosomal compartments, Sph causes changes in ion signaling and membrane trafficking, leading to the development of Niemann-Pick disease type C. Little is known about Sph interaction with other lipids in biological membranes; however, understanding the effect of Sph in the physical state of membranes might provide insights into its mode of action. Using complementary established fluorescence approaches, we show that Sph accumulation leads to the formation of Sph-enriched gel domains in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and POPC/sphingomyelin (SM)/cholesterol (Chol) model membranes. These domains are more easily formed in membrane models mimicking the neutral pH plasma membrane environment (PM) as compared to the acidic lysosomal membrane environment (LM), where higher Sph concentrations (or lower temperatures) are required. Electrophoretic light scattering measurements further revealed that in PM-raft models (POPC/SM/Chol), Sph is mainly neutral, whereas in LM models, the positive charge of Sph leads to electrostatic repulsion, reducing the Sph ability to form gel domains. Thus, formation of Sph-enriched domains in cellular membranes might be strongly regulated by Sph charge.

  18. Acidic intracellular pH shift during Caenorhabditis elegans larval development

    SciTech Connect

    Wadsworth, W.G.; Riddle, D.L. )

    1988-11-01

    During recovery from the developmentally arrested, nonfeeding dauer stage of the nemotode Caenorhabditis elegans, metabolic activation is accompanied by a decrease in intracellular pH (pH{sub i}). Phosphorus-31 nuclear magnetic resonance ({sup 31}P NMR) analyses of perchloric acid extracts show that inorganic phosphate predominates in dauer larvae, whereas ATP and other high-energy metabolites are abundant within 6 hr after dauer larvae have been placed in food to initiate development. Although metabolic activation has been associated with an alkaline pH{sub i} shift in other organisms, in vivo {sup 31}P NMR analysis of recovering dauer larvae shows a pH{sub i} decrease from {approx} 7.3 to {approx} 6.3 within 3 hr after the animals encounter food. This shift occurs before feeding begins, and it coincides with, or soon follows, the developmental commitment to recover from the dauer stage, suggesting that control of pH{sub i} may be important in the regulation of larval development in nematodes.

  19. Nucleophilic and general acid catalysis at physiological pH by a designed miniature esterase.

    PubMed

    Nicoll, Andrew J; Allemann, Rudolf K

    2004-08-07

    A 31-residue peptide (Art-Est) was designed to catalyse the hydrolysis of p-nitrophenyl esters through histidine catalysis on the solvent exposed face of the alpha-helix of bovine pancreatic polypeptide. NMR spectroscopy indicated that Art-Est adopted a stable 3-dimensional structure in solution. Art-Est was an efficient catalyst with second order rate constants of up to 0.050 M(-1) s(-1). The activity of Art-Est was a consequence of the increased nucleophilicity of His-22, which had a reduced pK(a) value of 5.5 as a consequence of its interaction with His-18 and the positively charged Arg-25 and Arg-26. Mass spectrometry and NMR spectroscopy confirmed that the Art-Est catalysed hydrolysis of p-nitrophenyl esters proceeded through an acyl-enzyme intermediate. A solvent kinetic isotope effect of 1.8 indicated that the transition state preceding the acyl intermediate was stabilised through interaction with the protonated side-chain of His-18 and indicated a reaction mechanism similar to that generally observed for natural esterases. The involvement in the reaction of two histidine residues with different pK(a) values led to a bell-shaped dependence of the reaction rate on the pH of the solution. The catalytic behaviour of Art-Est indicated that designed miniature enzymes can act in a transparent mechanism based fashion with enzyme-like behaviour through the interplay of several amino acid residues.

  20. Bioenergetics of serotonin transport by membrane vesicles derived from platelet dense granules.

    PubMed

    Fishkes, H; Rudnick, G

    1982-05-25

    A population of membrane vesicles derived from platelet-dense granules was isolated from platelet osmotic lysates by density gradient centrifugation. The preparation is relatively free from contamination by mitochondrial and soluble enzymatic activities and contains only traces of serotonin and adenine nucleotides, but retains significant amounts of Na+-dependent serotonin transport (plasma membrane) activity. In the presence of ATP and in the absence of Na+, these vesicles accumulate serotonin to intravesicular concentrations over 100 times that of the medium. Transport is a saturable process (Km = 1.15 microM) inhibited by reserpine, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, ammonia, and nigericin (in the presence of external potassium), but not by imipramine. When diluted into assay medium at pH 8.5, the interior of these vesicles remains relatively acidic. Addition of ATP further induced generation of a membrane potential (interior positive). The vesicles accumulate maximal concentrations of serotonin only when the vesicle interior is both acidic and positive with respect to the medium. These results are consistent with a transporter which catalyzes countertransport of at least two protons for each serotonin cation accumulated.

  1. Method for loading lipid like vesicles with drugs of other chemicals

    DOEpatents

    Mehlhorn, R.J.

    1998-06-09

    A method for accumulating drugs or other chemicals within synthetic, lipid-like vesicles by means of a pH gradient imposed on the vesicles just prior to use is described. The method is suited for accumulating molecules with basic or acid moieties which are permeable to the vesicles membranes in their uncharged form and for molecules that contain charge moieties that are hydrophobic ions and can therefore cross the vesicle membranes in their charged form. The method is advantageous over prior art methods for encapsulating biologically active materials within vesicles in that is achieves very high degrees of loading with simple procedures that are economical and require little technical expertise, furthermore kits which can be stored for prolonged periods prior to use without impairment of the capacity to achieve drug accumulation are described. A related application of the method consists of using this technology to detoxify animals that have been exposed to poisons with basic, weak acid or hydrophobic charge groups within their molecular structures. 2 figs.

  2. Method for loading lipid like vesicles with drugs of other chemicals

    DOEpatents

    Mehlhorn, Rolf Joachim

    1998-01-01

    A method for accumulating drugs or other chemicals within synthetic, lipid-like vesicles by means of a pH gradient imposed on the vesicles just prior to use is described. The method is suited for accumulating molecules with basic or acid moieties which are permeable to the vesicles membranes in their uncharged form and for molecules that contain charge moieties that are hydrophobic ions and can therefore cross the vesicle membranes in their charged form. The method is advantageous over prior art methods for encapsulating biologically active materials within vesicles in that is achieves very high degrees of loading with simple procedures that are economical and require little technical expertise, furthermore kits which can be stored for prolonged periods prior to use without impairment of the capacity to achieve drug accumulation are described. A related application of the method consists of using this technology to detoxify animals that have been exposed to poisons with basic, weak acid or hydrophobic charge groups within their molecular structures.

  3. Acid-coated Textiles (pH 5.5-6.5)--a New Therapeutic Strategy for Atopic Eczema?

    PubMed

    Jaeger, Teresa; Rothmaier, Markus; Zander, Holger; Ring, Johannes; Gutermuth, Jan; Anliker, Mark D

    2015-07-01

    Increased transepidermal water loss (TEWL) and decreased skin capacitance are characteristic features of the disturbed epidermal barrier in atopic eczema (AE). The "acid mantle", which is a slightly acidic film on the surface of the skin has led to the development of acidic emollients for skin care. In this context, the effect of citric acid-coated textiles on atopic skin has not been examined to date. A textile carrier composed of cellulose fibres was coated with a citric acid surface layer by esterification, ensuring a constant pH of 5.5-6.5. Twenty patients with AE or atopic diathesis were enrolled in the study. In a double-blind, half-side experiment, patients had to wear these textiles for 12 h a day for 14 days. On day 0 (baseline), 7 and 14, tolerability (erythema, pruritus, eczema, wearing comfort) and efficacy on skin barrier were assessed by TEWL skin hydration (corneometry/capacitance), pH and clinical scoring of eczema (SCORAD). Citric acid-coated textiles were well tolerated and improved eczema and objective parameters of skin physiology, including barrier function and a reduced skin surface pH, with potential lower pathogenic microbial colonisation.

  4. Tyramine biosynthesis is transcriptionally induced at low pH and improves the fitness of Enterococcus faecalis in acidic environments.

    PubMed

    Perez, Marta; Calles-Enríquez, Marina; Nes, Ingolf; Martin, Maria Cruz; Fernandez, Maria; Ladero, Victor; Alvarez, Miguel A

    2015-04-01

    Enterococcus faecalis is a commensal bacterium of the human gut that requires the ability to pass through the stomach and therefore cope with low pH. E. faecalis has also been identified as one of the major tyramine producers in fermented food products, where they also encounter acidic environments. In the present work, we have constructed a non-tyramine-producing mutant to study the role of the tyramine biosynthetic pathway, which converts tyrosine to tyramine via amino acid decarboxylation. Wild-type strain showed higher survival in a system that mimics gastrointestinal stress, indicating that the tyramine biosynthetic pathway has a role in acid resistance. Transcriptional analyses of the E. faecalis V583 tyrosine decarboxylase cluster showed that an acidic pH, together with substrate availability, induces its expression and therefore the production of tyramine. The protective role of the tyramine pathway under acidic conditions appears to be exerted through the maintenance of the cytosolic pH. Tyramine production should be considered important in the adaptability of E. faecalis to acidic environments, such as fermented dairy foods, and to survive passage through the human gastrointestinal tract.

  5. Acidification of endocytic vesicles by an ATP-dependent proton pump

    PubMed Central

    1983-01-01

    One of the early events in the pathway of receptor-mediated endocytosis is the acidification of the newly formed endocytic vesicle. To examine the mechanism of acidification, we used fluorescein-labeled alpha 2- macroglobulin (F-alpha 2M) as a probe for endocytic vesicle pH. Changes in pH were determined from the change in fluorescein fluorescence at 490-nm excitation as measured with a microscope spectrofluorometer. After endocytosis of F-alpha 2M, mouse fibroblast cells were permeabilized by brief exposure to the detergent digitonin. Treatment with the ionophore monensin or the protonophore carbonyl cyanide p- trifluoromethoxyphenylhydrazone (FCCP) caused a rapid increase in the pH of the endocytic vesicle. Upon removal of the ionophore, the endocytic vesicle rapidly acidified only when MgATP or MgGTP was added. Neither ADP nor the nonhydrolyzable analog, adenosine 5'-(beta, gamma- imido)triphosphate (AMP-PNP) could support acidification. The ATP- dependent acidification did not require a specific cation or anion in the external media. Acidification was insensitive to vanadate and amiloride but was inhibited by Zn2+ and the anion transport inhibitor diisothiocyanostilbene disulfonic acid (DIDS). We also examined the acidification of lysosomes with the permeabilized cell system, using fluorescein isothiocyanate dextran as probe. DIDS inhibited the ATP- dependent reacidification of lysosomes, although at a lower concentration than that for inhibition of endocytic vesicle reacidification. These results demonstrate that endocytic vesicles contain an ATP-dependent acidification mechanism that shares similar characteristics with the previously described lysosomal proton pump. PMID:6224803

  6. Denitrification potential in stream sediments impacted by acid mine drainage: effects of pH, various electron donors, and iron.

    PubMed

    Baeseman, J L; Smith, R L; Silverstein, J

    2006-02-01

    Acid mine drainage (AMD) contaminates thousands of kilometers of stream in the western United States. At the same time, nitrogen loading to many mountain watersheds is increasing because of atmospheric deposition of nitrate and increased human use. Relatively little is known about nitrogen cycling in acidic, heavy-metal-laden streams; however, it has been reported that one key process, denitrification, is inhibited under low pH conditions. The objective of this research was to investigate the capacity for denitrification in acidified streams. Denitrification potential was assessed in sediments from several Colorado AMD-impacted streams, ranging from pH 2.60 to 4.54, using microcosm incubations with fresh sediment. Added nitrate was immediately reduced to nitrogen gas without a lag period, indicating that denitrification enzymes were expressed and functional in these systems. First-order denitrification potential rate constants varied from 0.046 to 2.964 day(-1). The pH of the microcosm water increased between 0.23 and 1.49 pH units during denitrification. Additional microcosm studies were conducted to examine the effects of initial pH, various electron donors, and iron (added as ferrous and ferric iron). Decreasing initial pH decreased denitrification; however, increasing pH had little effect on denitrification rates. The addition of ferric and ferrous iron decreased observed denitrification potential rate constants. The addition of glucose and natural organic matter stimulated denitrification potential. The addition of hydrogen had little effect, however, and denitrification activity in the microcosms decreased after acetate addition. These results suggest that denitrification can occur in AMD streams, and if stimulated within the environment, denitrification might reduce acidity.

  7. Denitrification potential in stream sediments impacted by acid mine drainage: Effects of pH, various electron donors, and iron

    USGS Publications Warehouse

    Baeseman, J.L.; Smith, R.L.; Silverstein, J.

    2006-01-01

    Acid mine drainage (AMD) contaminates thousands of kilometers of stream in the western United States. At the same time, nitrogen loading to many mountain watersheds is increasing because of atmospheric deposition of nitrate and increased human use. Relatively little is known about nitrogen cycling in acidic, heavy-metal-laden streams; however, it has been reported that one key process, denitrification, is inhibited under low pH conditions. The objective of this research was to investigate the capacity for denitrification in acidified streams. Denitrification potential was assessed in sediments from several Colorado AMD-impacted streams, ranging from pH 2.60 to 4.54, using microcosm incubations with fresh sediment. Added nitrate was immediately reduced to nitrogen gas without a lag period, indicating that denitrification enzymes were expressed and functional in these systems. First-order denitrification potential rate constants varied from 0.046 to 2.964 day-1. The pH of the microcosm water increased between 0.23 and 1.49 pH units during denitrification. Additional microcosm studies were conducted to examine the effects of initial pH, various electron donors, and iron (added as ferrous and ferric iron). Decreasing initial pH decreased denitrification; however, increasing pH had little effect on denitrification rates. The addition of ferric and ferrous iron decreased observed denitrification potential rate constants. The addition of glucose and natural organic matter stimulated denitrification potential. The addition of hydrogen had little effect, however, and denitrification activity in the microcosms decreased after acetate addition. These results suggest that denitrification can occur in AMD streams, and if stimulated within the environment, denitrification might reduce acidity. ?? Springer Science+Business Media, Inc. 2006.

  8. Studies on tolfenamic acid-chitosan intermolecular interactions: effect of pH, polymer concentration and molecular weight.

    PubMed

    Ahmed, Sofia; Sheraz, Muhammad Ali; Rehman, Ihtesham Ur

    2013-06-01

    Solid-state properties of tolfenamic acid (TA) and its complexes with chitosan (CT) have been studied. Effect of medium pH, molecular weight of polymer and its different concentrations on these TA-CT complexes were studied in detail. Low and medium molecular weight CT have been used in different ratios at pH ranging from 4 to 6 and freeze-drying technique has been employed to modify the appearance of crystalline TA. Physical properties of the formed complexes have been studied by employing X-ray diffraction, differential scanning calorimetry and scanning electron microscopy; chemical structure has been studied using Fourier transform infrared spectroscopy. The results showed that both forms of the polymer exhibited complete conversion in 1:8 ratio at pH 4, 1:4 at pH 5 and 1:1 at pH 6 indicating a marked effect of pH on drug-polymer complexation. The percent crystallinity calculations indicated low molecular weight CT slightly more effective than the other form. No changes in the complexes have been observed during the 12 week storage under controlled conditions. Both forms of CT at different pH values indicated retardation of recrystallization in TA during cooling of the melt from 1:1 ratios exhibiting formation of strong intermolecular hydrogen bonding between the drug and the polymer.

  9. EFFECTS OF PH, SOLID/SOLUTION RATIO, IONIC STRENGTH, AND ORGANIC ACIDS ON PB AND CD SOPRTION ON KAOLINITE

    EPA Science Inventory

    Potentiometric and ion-selective electrode titrations together with batch sorption/desorption experiments, were performed to explain the aqueous and surface complexation reactions between kaolinite, Pb, Cd and three organic acids. Variables included pH, ionic strength, metal conc...

  10. Significance of pH on the Cytotoxic Potential of the Water Disinfection By-Product Iodoacetic Acid

    EPA Science Inventory

    Significance of pH on the Cytotoxic Potential of the Water Disinfection By-Product Iodoacetic Acid Vicki Richardson1, Susan D. Richardson2, Mary Moyer3, Jane Ellen Simmons1, and Anthony DeAngelo1, 1U.S. Environmental Protection Agency, Research Triangle Park, NC, 2University of...

  11. EFFECTS OF PH, SOLID/SOLUTION RATIO, IONIC STRENGTH, AND ORGANIC ACIDS ON PB AND CD SOPRTION ON KAOLINITE

    EPA Science Inventory

    Potentiometric and ion-selective electrode titrations together with batch sorption/desorption experiments, were performed to explain the aqueous and surface complexation reactions between kaolinite, Pb, Cd and three organic acids. Variables included pH, ionic strength, metal conc...

  12. Significance of pH on the Cytotoxic Potential of the Water Disinfection By-Product Iodoacetic Acid

    EPA Science Inventory

    Significance of pH on the Cytotoxic Potential of the Water Disinfection By-Product Iodoacetic Acid Vicki Richardson1, Susan D. Richardson2, Mary Moyer3, Jane Ellen Simmons1, and Anthony DeAngelo1, 1U.S. Environmental Protection Agency, Research Triangle Park, NC, 2University of...

  13. Embryonic common snapping turtles (Chelydra serpentina) preferentially regulate intracellular tissue pH during acid-base challenges.

    PubMed

    Shartau, Ryan B; Crossley, Dane A; Kohl, Zachary F; Brauner, Colin J

    2016-07-01

    The nests of embryonic turtles naturally experience elevated CO2 (hypercarbia), which leads to increased blood PCO2  and a respiratory acidosis, resulting in reduced blood pH [extracellular pH (pHe)]. Some fishes preferentially regulate tissue pH [intracellular pH (pHi)] against changes in pHe; this has been proposed to be associated with exceptional CO2 tolerance and has never been identified in amniotes. As embryonic turtles may be CO2 tolerant based on nesting strategy, we hypothesized that they preferentially regulate pHi, conferring tolerance to severe acute acid-base challenges. This hypothesis was tested by investigating pH regulation in common snapping turtles (Chelydra serpentina) reared in normoxia then exposed to hypercarbia (13 kPa PCO2 ) for 1 h at three developmental ages: 70% and 90% of incubation, and yearlings. Hypercarbia reduced pHe but not pHi, at all developmental ages. At 70% of incubation, pHe was depressed by 0.324 pH units while pHi of brain, white muscle and lung increased; heart, liver and kidney pHi remained unchanged. At 90% of incubation, pHe was depressed by 0.352 pH units but heart pHi increased with no change in pHi of other tissues. Yearlings exhibited a pHe reduction of 0.235 pH units but had no changes in pHi of any tissues. The results indicate common snapping turtles preferentially regulate pHi during development, but the degree of response is reduced throughout development. This is the first time preferential pHi regulation has been identified in an amniote. These findings may provide insight into the evolution of acid-base homeostasis during development of amniotes, and vertebrates in general.

  14. Dynamics of pH modification of an acidic protein bait used for tropical fruit flies (Diptera: Tephritidae).

    PubMed

    Heath, Robert R; Vazquez, Aime; Schnell, Elena Q; Villareal, Janett; Kendra, Paul E; Epsky, Nancy D

    2009-12-01

    Several species of Anastrepha and Bactrocera fruit flies (Diptera: Tephritidae) are captured in traps baited with the protein bait NuLure combined with borax (sodium tetraborate decahydrate) in an aqueous solution, typically 9% NuLure (vol:vol) with 3% borax (wt:vol). NuLure is an acid hydrolysate of corn and has an acidic pH. Addition of borax makes the solution more alkaline, and increase in alkalinity results in increase of ammonia release from the bait solution. This is a very dynamic system, with resultant pH affected by factors such as the amount of borax added, the pH of the water used for preparation, the age of the bait solution, and the development of microbial growth. Problems with borax include amount needed to increase alkalinity of NuLure solutions, which creates difficulties in disposing of spent bait in fruit fly trapping programs. Therefore, research was conducted to evaluate NaOH as an alternative method to increase alkalinity of NuLure solutions. Laboratory experiments compared effect of NaOH versus borax for pH modification on changes in pH and ammonia content of NuLure solutions over time. Although NuLure/NaOH solutions could be adjusted to a more alkaline pH than NuLure/borax solutions, borax plays a critical role in pH stability over time. However, the pH of NuLure/NaOH is stabilized when propylene glycol (10% vol:vol) was used to prepare the bait solution. The use of NaOH can provide an alternative to the use of borax to increase bait solution alkalinity.

  15. Effect of organic acids and temperature on survival of Shigella flexneri in broth at pH 4.

    PubMed

    Zaika, Laura L

    2002-09-01

    The survival of bacterial pathogens in acidified foods depends not only on the hydrogen ion concentration, but also on the type of acid and the storage temperature. Shigella flexneri is a foodborne pathogen that is acid tolerant. The survival of S. flexneri 5348 in brain heart infusion broth supplemented with 0.04 M acetic, citric, lactic, malic, or tartaric acid and adjusted to pH 4 with HCI or NaOH was studied. The control medium was brain heart infusion broth adjusted to pH 4 with HCI. Stationary-phase cells were inoculated into media at initial populations of 6 to 7 log10 CFU/ml and incubated at 4, 19, 28, and 37 degrees C. A two-phase linear inactivation model was applied to plate count data to derive lag times (tL) and slopes of the curves, from which D-values and time required for a 4-log10 decrease in population (T4D) were calculated. In all cases, survival increased with decreasing temperature. For each acid, tL, the D-value, and T4D increased with decreasing temperature. All acids inhibited S. flexneri to some extent but to differing degrees as follows: lactic acid, acetic acid > citric acid, malic acid, tartaric acid > HCl. The T4D values for the control medium and for media containing acetic, citric, lactic, malic, and tartaric acids were 64, 47, 50, 34, 58, and 52 h, respectively, at 37 degrees C and 2,607, 1,498, 1,905, 1,346, 1,726, and 2,134 h, respectively, at 4 degrees C. The results of this study indicate that organic acids may aid in the inactivation of Shigella. However, these data also suggest that foods stored at or below room temperature containing low levels (< 1%) of acids could cause illness if contaminated with Shigella.

  16. Linoleate isomerase activity occurs in lactic acid bacteria strains and is affected by pH and temperature.

    PubMed

    Gorissen, L; Weckx, S; Vlaeminck, B; Raes, K; De Vuyst, L; De Smet, S; Leroy, F

    2011-09-01

    To investigate the ability of lactic acid bacteria (LAB) to convert linoleic acid (LA) and α-linolenic acid (α-LNA) to conjugated linoleic acid (CLA) and conjugated linolenic acid (CLNA), respectively. To assess pH and temperature influences on CLA and CLNA production by Lactobacillus sakei LMG 13558. A screening of 48 LAB yielded one Lactobacillus curvatus, five Lactobacillus plantarum and four Lact. sakei strains displaying linoleate isomerase (LAI) activity. CLNA conversion percentages varied largely (1-60%). CLA conversion, occurring in three strains, was lower (2-5%). The LAI gene sequences of the ten LAI-positive strains shared 75-99% identity with the LAI gene sequence of a Lact. plantarum AS1.555. At pH 6.2, CLA and CLNA production by Lact. sakei LMG 13558 was higher at 30°C than at 20 and 25°C. At pH 5.5 (30°C) or 37°C (pH 6.2), LA was not converted and α-LNA only slightly converted. LAB show strain-dependent LAI activity. Production of CLA and CLNA is affected by pH and temperature, as shown for Lact. sakei LMG 13558. Several LAB produce CLA and/or CLNA, as shown for Lact. sakei and Lact. curvatus for the first time. These findings offer potential for the manufacturing of fermented functional foods. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  17. Oligomeric rearrangement of tick-borne encephalitis virus envelope proteins induced by an acidic pH.

    PubMed Central

    Allison, S L; Schalich, J; Stiasny, K; Mandl, C W; Kunz, C; Heinz, F X

    1995-01-01

    The flavivirus envelope protein E undergoes irreversible conformational changes at a mildly acidic pH which are believed to be necessary for membrane fusion in endosomes. In this study we used a combination of chemical cross-linking and sedimentation analysis to show that the envelope proteins of the flavivirus tick-borne encephalitis virus also change their oligomeric structure when exposed to a mildly acidic environment. Under neutral or slightly alkaline conditions, protein E on the surface of native virions exists as a homodimer which can be isolated by solubilization with the nonionic detergent Triton X-100. Solubilization with the same detergent after pretreatment at an acidic pH, however, yielded homotrimers rather than homodimers, suggesting that exposure to an acidic pH had induced a simultaneous weakening of dimeric contacts and a strengthening of trimeric ones. The pH threshold for the dimer-to-trimer transition was found to be 6.5. Because the pH dependence of this transition parallels that of previously observed changes in the conformation and hydrophobicity of protein E and that of virus-induced membrane fusion, it appears likely that the mechanism of fusion with endosomal membranes involves a specific rearrangement of the proteins in the viral envelope. Immature virions in which protein E is associated with the uncleaved precursor (prM) of the membrane protein M did not undergo a low-pH-induced rearrangement. This is consistent with a protective role of protein prM for protein E during intracellular transport of immature virions through acidic compartments of the trans-Golgi network. PMID:7529335

  18. Addition of formic acid or starter cultures to liquid feed. Effect on pH, microflora composition, organic acid concentration and ammonia concentration.

    PubMed

    Canibe, N; Miquel, N; Miettinen, H; Jensen, B B

    2001-01-01

    Some of the charateristics of good quality fermented liquid feed (FLF) are low pH, high numbers of lactic acid bacteria, and low numbers of enterobacteria. In order to test strategies to avoid a proliferation of enterobacteria during the initial phase of FLF elaboration, two in vitro studies were carried out. Addition of various doses of formic acid or two different starter cultures were tested. Adding 0.1% formic acid or L. plantarum VTT E-78076 to the liquid feed seemed to be addecuate ways of inhibiting the growth of enterobacteria, without depleting the growth of lactic acid bacteria.

  19. pH-sensitive vesicles based on a biocompatible zwitterionic diblock copolymer.

    PubMed

    Du, Jianzhong; Tang, Yiqing; Lewis, Andrew L; Armes, Steven P

    2005-12-28

    Highly biocompatible pH-sensitive diblock copolymer vesicles were prepared from the self-assembly of a biocompatible zwitterionic copolymer, poly[2-(methacryloyloxy)ethyl phosphorylcholine-block-2-(diisopropylamino)ethyl methacrylate], PMPC-b-PDPA. Vesicle formation occurred spontaneously by adjusting the solution pH from pH 2 to above 6, with the hydrophobic PDPA chains forming the vesicle walls. Transmission electron microscopy (TEM), dynamic laser light scattering (DLS), and UV-visible absorption spectrophotometry were used to characterize these vesicles. Gold nanoparticle-decorated vesicles were also obtained by treating the vesicles with HAuCl4, followed by NaBH4.

  20. Effect of low pH start-up on continuous mixed-culture lactic acid fermentation of dairy effluent.

    PubMed

    Choi, Gyucheol; Kim, Jaai; Lee, Changsoo

    2016-12-01

    Mixed-culture fermentation that does not require an energy-intensive sterilization process is a viable approach for the economically feasible production of lactic acid (LA) due to the potential use of organic waste as feedstock. This study investigated mixed-culture LA fermentation of whey, a high-strength organic wastewater, in continuous mode. Variations in the hydraulic retention time (HRT) from 120 to 8 h under different pH regimes in two thermophilic reactors (55 °C) were compared for their fermentation performance. One reactor was maintained at a low pH (pH 3.0) during operation at HRTs of 120 to 24 h and then adjusted to pH 5.5 in the later phases of fermentation at HRTs of 24 to 8 h (R1), while the second reactor was maintained at pH 5.5 throughout the experiment (R2). Although the LA production in R1 was negligible at low pH, it increased dramatically after the pH was raised to 5.5 and exceeded that in R2 when stabilized at HRTs of 8 and 12 h. The maximum yield (0.62 g LA/g substrate fed as the chemical oxygen demand (COD) equivalent), the production rate (11.5 g/L day), and the selectivity (95 %) of LA were all determined at a 12-h HRT in R1. Additionally, molecular and statistical analyses revealed that changes in the HRT and the pH significantly affected the bacterial community structure and thus the fermentation characteristics of the experimental reactors. Bacillus coagulans was likely the predominant LA producer in both reactors. The overall results suggest that low pH start-up has a positive effect on yield and selectivity in mixed-culture LA fermentation.

  1. Metabolic flux modeling of detoxification of acetic acid by Ralstonia eutropha at slightly alkaline pH levels.

    PubMed

    Yu, J; Wang, J

    2001-06-20

    Ralstonia eutropha grows on and produces polyhydroxyalkanoates (PHAs) from fermentation acids. Acetic acid, one major organic acid from acidogenesis of organic wastes, has an inhibitory effect on the bacterium at slightly alkaline pH (6 g HAc/L at pH 8). The tolerance of R. eutropha to acetate, however, was increased significantly up to 15 g/L at the slightly alkaline pH level with high cell mass concentration. A metabolic cell model with five fluxes is proposed to depict the detoxification mechanism including mass transfer and acetyl-CoA formation of acetic acid and the formation of three final metabolic products, polyhydroxybutyrate (PHB), active biomass, and CO(2). The fluxes were measured under different conditions such as cell mass concentration, acetic acid concentration, and medium composition. The experimental results indicate that the acetate detoxification by high cell mass concentration is attributed to the increased fluxes at high extracellular acetate concentrations. The fluxes could be doubled to reduce and hence detoxify the accumulated intracellular acetate anions.

  2. Surface-enhanced Raman spectroscopy study on the structure changes of 4-Mercaptophenylboronic Acid under different pH conditions

    NASA Astrophysics Data System (ADS)

    Su, Hongyang; Wang, Yue; Yu, Zhi; Liu, Yawen; Zhang, Xiaolei; Wang, Xiaolei; Sui, Huimin; Sun, Chengbin; Zhao, Bing

    2017-10-01

    4-Mercaptophenylboronic Acid (4-MPBA) plays pivotal role in various fields. The orientation and existing form of the 4-MPBA strongly depend on the pH value of the media. The general aim of this work is to obtain information about the structure changes of 4-MPBA absorbed on Ag nanoparticles in different pH environment. Surface-enhanced Raman spectroscopy (SERS) technique is a simple and rapid method to study adsorption phenomena at molecule level. The investigation is done by means of SERS. In order to interpret the experimental information, a series of SERS spectra is carried out. The relative intensities of the totally symmetric (a1 mode) and non-totally symmetric (b2 mode) bands in the SERS spectra of 4-MPBA change depend on the environmental pH values, which is a manifestation of charge transfer (CT) processes. The degree of charge transfer increases with the pH value of the media changing from acidity to alkalinity. The structure changes of MPBA had been carried out in different pH environment. We envision that this approach will be of great significance in related fields of 4-MPBA-involved detection.

  3. Extracellular pH regulation in microdomains of colonic crypts: effects of short-chain fatty acids.

    PubMed Central

    Chu, S; Montrose, M H

    1995-01-01

    It has been suggested that transepithelial gradients of short-chain fatty acids (SCFAs; the major anions in the colonic lumen) generate pH gradients across the colonic epithelium. Quantitative confocal microscopy was used to study extracellular pH in mouse distal colon with intact epithelial architecture, by superfusing tissue with carboxy SNARF-1 (a pH-sensitive fluorescent dye). Results demonstrate extracellular pH regulation in two separate microdomains surrounding colonic crypts: the crypt lumen and the subepithelial tissue adjacent to crypt colonocytes. Apical superfusion with (i) a poorly metabolized SCFA (isobutyrate), (ii) an avidly metabolized SCFA (n-butyrate), or (iii) a physiologic mixture of acetate/propionate/n-butyrate produced similar results: alkalinization of the crypt lumen and acidification of subepithelial tissue. Effects were (i) dependent on the presence and orientation of a transepithelial SCFA gradient, (ii) not observed with gluconate substitution, and (iii) required activation of sustained vectorial acid/base transport by SCFAs. Results suggest that the crypt lumen functions as a pH microdomain due to slow mixing with bulk superfusates and that crypts contribute significant buffering capacity to the lumen. In conclusion, physiologic SCFA gradients cause polarized extracellular pH regulation because epithelial architecture and vectorial transport synergize to establish regulated microenvironments. Images Fig. 1 Fig. 3 PMID:7724557

  4. Sorption of triclosan onto activated carbon, kaolinite and montmorillonite: effects of pH, ionic strength, and humic acid.

    PubMed

    Behera, Shishir Kumar; Oh, Seok-Young; Park, Hung-Suck

    2010-07-15

    Sorption of triclosan on three sorbents, viz., activated carbon, kaolinite and montmorillonite was studied as a function of pH, ionic strength and humic acid (HA) concentration through controlled batch experiments. Triclosan sorption was found to be higher in the acidic pH range, as varying pH showed significant influence on the surface charge of the sorbents and degree of ionization of the sorbate. Sorption capacity of the sorbents increased with an increase in the ionic strength of solution. At low pH (pH 3), the overall increase in triclosan sorption was 1.2, approximately 4 and 3.5 times, respectively for activated carbon, kaolinite and montmorillonite when ionic strength was increased from 1x10(-3) to 5x10(-1) M. Triclosan sorption onto activated carbon decreased from 31.4 to 10.6 mg g(-1) by increasing the HA concentration to 200 mg C L(-1). However, during sorption onto kaolinite and montmorillonite, the effect of HA was very complex probably due to (i) hydrophobicity (log K(ow)=4.76) of triclosan; and (ii) complexation of HA with triclosan. Though triclosan sorption onto activated carbon is higher, the potential of kaolinite and montmorillonite in controlling the transport of triclosan in subsurface environment can still be appreciable.

  5. The Acid Test of Fluoride: How pH Modulates Toxicity

    PubMed Central

    Sharma, Ramaswamy; Tsuchiya, Masahiro; Skobe, Ziedonis; Tannous, Bakhos A.; Bartlett, John D.

    2010-01-01

    Background It is not known why the ameloblasts responsible for dental enamel formation are uniquely sensitive to fluoride (F−). Herein, we present a novel theory with supporting data to show that the low pH environment of maturating stage ameloblasts enhances their sensitivity to a given dose of F−. Enamel formation is initiated in a neutral pH environment (secretory stage); however, the pH can fall to below 6.0 as most of the mineral precipitates (maturation stage). Low pH can facilitate entry of F− into cells. Here, we asked if F− was more toxic at low pH, as measured by increased cell stress and decreased cell function. Methodology/Principal Findings Treatment of ameloblast-derived LS8 cells with F− at low pH reduced the threshold dose of F− required to phosphorylate stress-related proteins, PERK, eIF2α, JNK and c-jun. To assess protein secretion, LS8 cells were stably transduced with a secreted reporter, Gaussia luciferase, and secretion was quantified as a function of F− dose and pH. Luciferase secretion significantly decreased within 2 hr of F− treatment at low pH versus neutral pH, indicating increased functional toxicity. Rats given 100 ppm F− in their drinking water exhibited increased stress-mediated phosphorylation of eIF2α in maturation stage ameloblasts (pH<6.0) as compared to secretory stage ameloblasts (pH∼7.2). Intriguingly, F−-treated rats demonstrated a striking decrease in transcripts expressed during the maturation stage of enamel development (Klk4 and Amtn). In contrast, the expression of secretory stage genes, AmelX, Ambn, Enam and Mmp20, was unaffected. Conclusions The low pH environment of maturation stage ameloblasts facilitates the uptake of F−, causing increased cell stress that compromises ameloblast function, resulting in dental fluorosis. PMID:20531944

  6. Using Dynamic Covalent Chemistry To Drive Morphological Transitions: Controlled Release of Encapsulated Nanoparticles from Block Copolymer Vesicles.

    PubMed

    Deng, Renhua; Derry, Matthew J; Mable, Charlotte J; Ning, Yin; Armes, Steven P

    2017-06-07

    Dynamic covalent chemistry is exploited to drive morphological order-order transitions to achieve the controlled release of a model payload (e.g., silica nanoparticles) encapsulated within block copolymer vesicles. More specifically, poly(glycerol monomethacrylate)-poly(2-hydroxypropyl methacrylate) (PGMA-PHPMA) diblock copolymer vesicles were prepared via aqueous polymerization-induced self-assembly in either the presence or absence of silica nanoparticles. Addition of 3-aminophenylboronic acid (APBA) to such vesicles results in specific binding of this reagent to some of the pendent cis-diol groups on the hydrophilic PGMA chains to form phenylboronate ester bonds in mildly alkaline aqueous solution (pH ∼ 10). This leads to a subtle increase in the effective volume fraction of this stabilizer block, which in turn causes a reduction in the packing parameter and hence induces a vesicle-to-worm (or vesicle-to-sphere) morphological transition. The evolution in copolymer morphology (and the associated sol-gel transitions) was monitored using dynamic light scattering, transmission electron microscopy, oscillatory rheology, and small-angle X-ray scattering. In contrast to the literature, in situ release of encapsulated silica nanoparticles is achieved via vesicle dissociation at room temperature; moreover, the rate of release can be fine-tuned by varying the solution pH and/or the APBA concentration. Furthermore, this strategy also works (i) for relatively thick-walled vesicles that do not normally exhibit stimulus-responsive behavior and (ii) in the presence of added salt. This novel molecular recognition strategy to trigger morphological transitions via dynamic covalent chemistry offers considerable scope for the design of new stimulus-responsive copolymer vesicles (and hydrogels) for targeted delivery and controlled release of cargoes. In particular, the conditions used in this new approach are relevant to liquid laundry formulations, whereby enzymes require

  7. The impact of 10% α-hydroxy acid emulsion on skin pH.

    PubMed

    Schreml, S; Meier, R J; Albert, M G; Seidl, U; Zeller, V; Behm, B; Landthaler, M; Abels, C; Babilas, P

    2012-01-01

    The effects of a 10% α-hydroxy acid (AHA) oil/water (O/W) emulsion on the pH of human skin surface (pH(ss)) and stratum corneum (SC; pH(sc)) were evaluated in vivo. The AHA O/W emulsion was applied to an area on the volar forearm of male volunteers (n = 12), and then wiped off after 10 min. Prior to application and over the following 3 h, the pH(ss) was measured. We used glass electrode measurements and time domain dual lifetime referencing (tdDLR) with luminescent sensor foils. In another experiment (n = 5), the impact of the AHA O/W emulsion on the pH(sc) gradient was measured by tape stripping of the SC of the volar forearm after application of the AHA O/W emulsion. Compared with pH(ss) values prior to treatment [5.2 ± 1.7 (tdDLR) or 4.8 ± 0.5 (electrode)], the pH(ss) was significantly reduced 10 min after application [4.0 ± 0.3 (tdDLR) or 4.1 ± 0.1 (electrode)] and the pH(ss) remained significantly reduced over the measurement period of 3 h [after 3 h: 4.4 ± 0.2 (tdDLR) or 4.5 ± 0.3 (electrode)]. The AHA O/W emulsion significantly reduced the pH(sc) even down to deep layers of the SC. After a 10-min application time, the 10% AHA O/W emulsion reduces the pH(ss) (for at least 3 h) and pH(sc) in deep layers of the SC. Copyright © 2011 S. Karger AG, Basel.

  8. Function of aspartic acid residues in optimum pH control of L-arabinose isomerase from Lactobacillus fermentum.

    PubMed

    Xu, Zheng; Li, Sha; Feng, Xiaohai; Zhan, Yijing; Xu, Hong

    2014-05-01

    L-Arabinose isomerase (L-AI) catalyzes the isomerization of L-arabinose to L-ribulose and D-galactose to D-tagatose. Most reported L-AIs exhibit neutral or alkaline optimum pH, which is less beneficial than acidophilic ones in industrial D-tagatose production. Lactobacillus fermentum L-AI (LFAI) is a thermostable enzyme that can achieve a high conversion rate for D-galactose isomerization. However, its biocatalytic activity at acidic conditions can still be further improved. In this study, we report the single- and multiple-site mutagenesis on LFAI targeting three aspartic acid residues (D268, D269, and D299). Some of the lysine mutants, especially D268K/D269K/D299K, exhibited significant optimum pH shifts (from 6.5 to 5.0) and enhancement of pH stability (half-life time increased from 30 to 62 h at pH 6.0), which are more favorable for industrial applications. With the addition of borate, D-galactose was isomerized into D-tagatose by D268K/D269K/D299K at pH 5.0, resulting in a high conversion rate of 62 %. Based on the obtained 3.2-Å crystal structure of LFAI, the three aspartic acid residues were found to be distant from the active site and possibly did not participate in substrate catalysis. However, they were proven to possess similar optimum pH control ability in other L-AI, such as that derived from Escherichia coli. This study sheds light on the essential residues of L-AIs that can be modified for desired optimum pH and better pH stability, which are useful in D-tagatose bioproduction.

  9. Relative effectiveness of various anions on the solubility of acidic Hypoderma lineatum collagenase at pH 7.2.

    PubMed Central

    Carbonnaux, C.; Ries-Kautt, M.; Ducruix, A.

    1995-01-01

    The effects of various anions on decreasing the solubility of acidic Hypoderma lineatum collagenase at pH 7.2 and 18 degrees C were qualitatively defined by replacing the crystallizing agent of known crystallization conditions by various ammonium salts. The solubility curves measured in the presence of the sulfate, phosphate, citrate, and chloride ammonium salts gave the following ranking of anions: HPO4(2-)/H2PO4- > SO4(2-) > citrate 3-/citrate2- >> Cl-. This order is in agreement with the Hofmeister series. In a previous study on the solubility at pH 4.5 of lysozyme, a basic protein, the effectiveness of anions in decreasing the solubility was found to be in the reverse order. This suggests that the effectiveness of anions in the crystallization of proteins is dependent on the net charge of the protein, i.e., depending on whether a basic protein is crystallized at acidic pH or an acidic protein at basic pH. PMID:8535249

  10. Investigating Effects of Acidic pH on Proliferation, Invasion and Drug-Induced Apoptosis in Lymphoblastic Leukemia.

    PubMed

    Bohloli, Mahbobeh; Atashi, Amir; Soleimani, Masoud; Kaviani, Saeid; Anbarlou, Azadeh

    2016-12-01

    Some studies have shown that extracellular pH in tumors, which results in tumor progression, is less than that in normal tissues. The aim of this study was to investigate the effects of extracellular acidic pH on proliferation, invasion, and drug-induced apoptosis in acute lymphoblastic cells. The cells were cultured in different pH (pH 6.6 and pH 7.4) for 12 days. Cell proliferation was assessed by MTT assay and cell invasion was assayed by invasion assay and gene expression analysis of MMP-9. Drug-induced apoptosis was evaluated after exposure to doxorubicin for 24 hours by annexin V/PI staining and gene expression analysis of BAX pro-apoptotic protein. The results indicated the enhanced growth and invasion of leukemic cells at pH 6.6 (P ≤ 0.05). Furthermore, the cells at pH 6.6 were resistant to apoptosis by doxorubicin (P ≤ 0.05). It can be concluded that acidic pH increases the proliferation, invasion and reduces the drug-induced apoptosis in acute lymphoblastic leukemia. Extracellular acidity can influence the behavior of leukemic cells and therefore, the manipulation of extracellular liquid can be selected as a therapeutic strategy for leukemia, especially for acute lymphoblastic leukemia.

  11. [Effects of solution pH and simulated acid rain on the behavior of two sulfonylurea herbicides in soil].

    PubMed

    Zhang, Wei; Wang, Jin-Jun

    2007-03-01

    By the methods of batch equilibration and leaching, this paper studied the effects of solution pH and simulated acid rain on the behavior of bensulfuron-methyl and metsulfuron-methyl in soil. The results showed that the adsorption isotherms of these two herbicides fitted Freundlich equation well, and their adsorbed amounts reduced obviously with the increasing pH of water-soil system, which in turn promoted the translocation of the herbicides in soil. The adsorption coefficient (Kf) was positively correlated with soil organic matter and clay contents, while negatively correlated with soil pH. The higher pH of simulated acid rain had a greater contribution on the leaching of the two sulfonylurea herbicides, and their leached amount was increased with increasing acid rain. There was a close relationship between the leaching of the herbicides and the properties of soil. The soils with higher contents of organic matter and clay had a greater retention capability to the herbicides.

  12. Calcium transport in sealed vesicles from red beet (Beta vulgaris L. ) storage tissue. II. Characterization of /sup 45/Ca/sup 2 +/ uptake into plasma membrane vesicles

    SciTech Connect

    Giannini, J.L.; Ruiz-Cristin, J.; Briskin, D.P.

    1987-12-01

    Calcium uptake was examined in sealed plasma membrane vesicles isolated from red beet (Beta vulgaris L.) storage tissue using /sup 45/Ca/sup 2 +/. Uptake of /sup 45/Ca/sup 2 +/ by the vesicles was ATP-dependent and radiotracer accumulated by the vesicles could be released by the addition of the calcium ionophore A23187. The uptake was stimulated by gramicidin D but slightly inhibited by carbonylcyanide m-chlorophenylhydrazone. Although the latter result might suggest some degree of indirect coupling of /sup 45/Ca/sup 2 +/ uptake to ATP utilization via ..delta mu..H/sup +/, no evidence for a secondary H/sup +//Ca/sup 2 +/ antiport in this vesicle system could be found. Following the imposition of an acid-interior pH gradient, proton efflux from the vesicle was not enhanced by the addition of Ca/sup 2 +/ and an imposed pH gradient could not drive /sup 45/Ca/sup 2 +/ uptake. Optimal uptake of /sup 45/Ca/sup 2 +/ occurred broadly between pH 7.0 and 7.5 and the transport was inhibited by orthovanadate, N,N'-dicyclohexylcarbodiimide, and diethylstilbestrol but insensitive to nitrate and azide. The dependence of /sup 45/Ca/sup 2 +/ uptake on both calcium and Mg:ATP concentration demonstrated saturation kinetics with K/sub m/ values of 6 micromolar and 0.37 millimolar, respectively. While ATP was the preferred substrate for driving /sup 45/Ca/sup 2 +/ uptake, GTP could drive transport at about 50% of the level observed for ATP. The results of this study demonstrate the presence of a unique primary calcium transport system associated with the plasma membrane which could drive calcium efflux from the plant cell.

  13. H/sup +/-ATPase in the maintenance of an acidic pH in lamellar bodies isolated from rat lungs

    SciTech Connect

    Fisher, A.B.; Chander, A.; Reicherter, J.

    1986-03-05

    The authors have previously shown that lung lamellar bodies maintain an acidic internal pH by an energy dependent mechanism. The pH of lamellar bodies isolated from rat lung was characterized by fluorescence microscopy with quinacrine HCl (8 ..mu..M) or by accumulation of (/sup 14/C) methylamine. Quinacrine fluorescence of lamellar bodies was decreased with 10mM NH/sub 4/Cl or by omission of ATP from the medium. Accumulation of (/sup 14/C)methylamine(inside/outside) at 2 min incubation was 8.1 +/- 0.47 (n=8) under control conditions in the presence of 10mM ATP.MgCl/sub 2/ (mean +/- SE; n=6) corresponding to a pH gradient of 0.91 +/- 0.03. Accumulation in the absence of ATP was 2.9 +/- 0.20 (pH gradient 0.47). Addition of nigericin (2..mu..g/ml) in the presence of ATP lowered the pH gradient to 0.49 +/- 0.051 (n=4) suggesting that the normal gradient was maintained by K/sup +//H/sup +/ exchange across the lamellar body membrane. Addition of inhibits of H/sup +/-ATPase, N-ethylmaleamide (NEM) (1mM) or N,N'-dicyclohexylcarbodiimide (DCCD) (0.02 mM) decreased the pH gradient to 0.47 +/- 0.11 (n=3) and 0.57 +/- 0.06 (n=3), respectively. These results suggest involvement of H/sup +/-ATPase in the maintenance of an acidic pH in lung lamellar bodies.

  14. Influence of metal ions and pH on the hydraulic properties of potential acid sulfate soils

    NASA Astrophysics Data System (ADS)

    Le, T. M. H.; Collins, R. N.; Waite, T. D.

    2008-07-01

    SummaryAcid sulfate soils (ASS) cover extensive areas of east Australian coastal floodplains. Upon oxidation, these hydromorphic pyritic sediments produce large quantities of sulfuric acid. In addition, due to their geographic location, these soils may also come in contact with high ionic strength estuarine tidal waters. As a result, there is typically a large variation in acidity (pH) and cation concentrations in soil porewaters and adjacent aquatic systems (e.g., agricultural field drains, rivers, estuaries, etc.). Acid sulfate soils, especially from the unoxidized gelatinous deeper layers, contain a relatively high proportion of montmorillonite, which is wellknown for its shrink-swell properties. Variations in cation concentrations, including H3O+, can influence montmorillonite platelet interactions and may, thus, also significantly affect the hydraulic conductivity of materials containing this clay. In this paper we report on the effect of four common cations, at reasonable environmental concentrations, on the hydraulic properties of potential (unoxidized) acid sulfate soil materials. The natural system was simplified by examining individually the effects of each cation (H+, Ca2+, Fe2+ and Na+) on a soil-water suspension in a filtration cell unit. Moisture ratio, hydraulic conductivity and the consolidation coefficient of the deposited filter cakes were calculated using material coordinates theory. The results indicate that the hydraulic conductivity of potential acid sulfate soils increases at low pH and with cation concentration. Although an increase in the charge of amphoteric edge groups on montmorillonite clays may result in some aggregation between individual clay platelets, we conclude that the extent of these changes are unlikely to cause significant increases in the transportation of acidity (and contaminants) through potential acid sulfate soils as the hydraulic conductivity of these materials remain low (<10-9 m/s) at pH and ionic conditions normally

  15. Potentiometric pH Measurements of Acidity Are Approximations, Some More Useful than Others

    ERIC Educational Resources Information Center

    de Levie, Robert

    2010-01-01

    A recent article by McCarty and Vitz "demonstrating that it is not true that pH = -log[H+]" is examined critically. Then, the focus shifts to underlying problems with the IUPAC definition of pH. It is shown how the potentiometric method can provide "estimates" of both the IUPAC-defined hydrogen activity "and" the hydrogen ion concentration, using…

  16. Potentiometric pH Measurements of Acidity Are Approximations, Some More Useful than Others

    ERIC Educational Resources Information Center

    de Levie, Robert

    2010-01-01

    A recent article by McCarty and Vitz "demonstrating that it is not true that pH = -log[H+]" is examined critically. Then, the focus shifts to underlying problems with the IUPAC definition of pH. It is shown how the potentiometric method can provide "estimates" of both the IUPAC-defined hydrogen activity "and" the hydrogen ion concentration, using…

  17. Demonstration of in situ product recovery of butyric acid via CO2 -facilitated pH swings and medium development in two-phase partitioning bioreactors.

    PubMed

    Peterson, Eric C; Daugulis, Andrew J

    2014-03-01

    Production of organic acids in solid-liquid two-phase partitioning bioreactors (TPPBs) is challenging, and highly pH-dependent, as cell growth occurs near neutral pH, while acid sorption occurs only at low pH conditions. CO2 sparging was used to achieve acidic pH swings, facilitating undissociated organic acid uptake without generating osmotic stress inherent in traditional acid/base pH control. A modified cultivation medium was formulated to permit greater pH reduction by CO2 sparging (pH 4.8) compared to typical media (pH 5.3), while still possessing adequate nutrients for extensive cell growth. In situ product recovery (ISPR) of butyric acid (pKa = 4.8) produced by Clostridium tyrobutyricum was achieved through intermittent CO2 sparging while recycling reactor contents through a column packed with absorptive polymer Hytrel® 3078. This polymer was selected on the basis of its composition as a polyether copolymer, and the use of solubility parameters for predicting solute polymer affinity, and was found to have a partition coefficient for butyric acid of 3. Total polymeric extraction of 3.2 g butyric acid with no CO2 mediated pH swings was increased to 4.5 g via CO2 -facilitated pH shifting, despite the buffering capacity of butyric acid, which resists pH shifting. This work shows that CO2 -mediated pH swings have an observable positive effect on organic acid extraction, with improvements well over 150% under optimal conditions in early stage fermentation compared to CO2 -free controls, and this technique can be applied other organic acid fermentations to achieve or improve ISPR.

  18. Tolerance of acid-adapted and non-adapted Escherichia coli O157:H7 cells to reduced pH as affected by type of acidulant.

    PubMed

    Deng, Y; Ryu, J H; Beuchat, L R

    1999-02-01

    A study was carried out to determine if three strains of Escherichia coli O157:H7 grown (18 h) in Tryptic Soy Broth (TSB) and TSB supplemented with 1.25% glucose (TSBG), i.e. unadapted and acid-adapted cells, respectively, exhibited changes in tolerance to reduced pH when plated on Tryptic Soy Agar (TSA) acidified (pH</