Lipid remodeling under acidic conditions and its interplay with low Pi stress in Arabidopsis

Plant Molecular Biology - Here we show that accumulation of galactose-containing lipids in plastid membranes in shoots and the other membranes in roots maintains Arabidopsis growth under acidic...     

Variability and its implications for host-parasite interactions

Variability in host-parasite interactions has considerable impact on the ecology and evolution of parasites and on the epidemiology of disease. The nature of the impact depends largely on the level of ecological organization where variability occurs: variability of parasites within their individual hosts, variability of host individuals within populations, or variability of hosts and parasites among populations. In this review, Paul Schmid-Hempel and Jacob Koella give some examples of variability at each of these levels, with particular emphasis on microparasites (defined broadly as viruses, bacteria and protozoa), consider the maintenance of the variability, and describe the implications of variability for the epidemiology of disease and the ecology of host parasite associations. In particular, they describe how variability at each level of ecological organization can affect the perception of AIDS and the evolution of virulence.     

Enhanced lactic acid bacteria viability with yeast coincubation under acidic conditions

ABSTRACT

The enhancing effects of yeasts on the viability of lactic acid bacteria (LAB) under acidic conditions were investigated. Meyerozyma guilliermondii, coaggregative with both LAB strains under acidic conditions, significantly enhanced the viability of Lactobacillus pentosus and L. paracasei in pH 3.0 lactic acid (LA) buffer at 10°C (p < 0.05). Non-coaggregative yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Cyberlindnera saturnus) also significantly enhanced the LAB viability (p < 0.05), and physical contact between LAB and yeasts was not essential for the viability-enhancing effect, indicating that the coaggregation had no relation to the enhancing mechanism. Although yeast metabolites and LA assimilation had no enhancing effect, hydrogen peroxide (H₂O₂) decreased after yeast coincubation, and H₂O₂ elimination improved L. pentosus viability. H₂O₂ elimination alone did not sufficiently improve L. paracasei viability, but the addition of antioxidants was effective. These results suggest that the antioxidant activity of yeast increased the LAB viability under acidic conditions.     

Mutagens formed from butylated hydroxyanisole treated with nitrite under acidic conditions

The reaction products from butylated hydroxyanisole treated with nitrite under acidic conditions were investigated for mutagenic activity in Salmonella typhimurium his reversion assay and for DNA-damaging activity using H17 Rec+ (wild) and M45 Rec- (recombinationless) of Bacillus subtilis. The chloroform extract of the reaction mixture showed 9 spots on thin-layer chromatography (TLC). Compounds from 2 spots on the TLC had high mutagenic activity in TA100 without S9 mix, with DNA-damaging activity. The 2 mutagens were then crystallized from the reaction mixture and identified to be 2-tert.-butyl-p-quinone (t-BQ) and the dimer of t-BQ; 3,3'-di-tert.-butyl-biphenyldiquinone-(2,5,2',5') (BBDQ), from their instrumental analysis. The mutagenic activities of t-BQ and BBDQ were determined by Ames test, and the induced mutation frequencies were about 1.9 X 10(-4) (t-BQ) and 8.3 X 10(-5) (BBDQ).     

Effects of pH on protein-protein interactions and implications for protein phase behavior

The effects of pH on protein interactions and protein phase behavior were investigated by measuring the reduced second osmotic virial coefficient (b2) for ovalbumin and catalase, and the aggregate and crystal solubilities for ovalbumin, beta-lactoglobulin A and B, ribonuclease A and lysozyme. The b2 trends observed for ovalbumin and catalase show that protein interactions become increasingly attractive with decreasing pH. This trend is in good agreement with ovalbumin phase behavior, which was observed to evolve progressively with decreasing pH, leading to formation of amorphous aggregates instead of gel bead-like aggregates, and spherulites instead of needle-like crystals. For both acidic and basic proteins, the aggregate solubility during protein salting-out decreased with decreasing pH, and contrary to what is commonly believed, neither aggregate nor crystal solubility had a minimum at the isoelectric point. beta-Lactoglobulin B was the only protein investigated to show salting-in behavior, and crystals were obtained at low salt concentrations in the vicinity of its isoelectric point. The physical origin of the different trends observed during protein salting-in and salting-out is discussed, and the implications for protein crystallization are emphasized.     

Stability and transformation of the glucocorticoid receptor under acidic conditions

The [3H]triamcinolone acetonide ([3H]TA)-binding ability of the rat liver glucocorticoid receptor (GR) was investigated under acidic conditions, ranging from pH 2 to 7.3. Both in the presence and absence of 10 mM molybdate, the [3H]TA-binding ability decreased below pH 6.5 and was almost completely lost below pH 5, pH 5.9 +/- 0.1 giving 50% [3H]TA-binding. The binding ability was recovered when the pH of the cytosol was reversed to 7.3 or the precipitate obtained on acidification was dissolved in a buffer of pH 7.3. Moreover, in the absence of molybdate, the [3H]TA-GR complexes formed at pH 7.3 remained unchanged until pH 5. Then they decreased, pH 3.9 +/- 0.1 giving 50% binding, and completely disappeared at pH 3. [3H]TA-binding activity recovered from the precipitate also decreased in a similar pH region (a 50% decrease in binding being observed at pH 4.2 +/- 0.04). These results suggest that rat liver GR is rather resistant under acidic conditions and that it exists in a peculiar state below pH 5.9 to approximately 4 as to its ligand binding property: unoccupied GR has no [3H]TA-binding ability but [3H]TA-GR complexes once formed at neutral pH do not dissociate. [3H]TA-GR complexes recovered from the precipitate at pH 5 had a Stokes radius of 7.5 nm, little DNA-cellulose-binding ability and sedimented at 8.6S on glycerol gradient centrifugation, indicating that the receptor existed in a nontransformed state. In addition, both occupied and unoccupied GR were transformed at about pH 4, their being 50% transformation. This transformation was accompanied by irreversible denaturation of the receptor.(ABSTRACT TRUNCATED AT 250 WORDS)     

Oxidation products of acylated anthocyanins under acidic and neutral conditions

Acylated anthocyanidin-3,5-diglucosides are oxidized with H₂O₂ under acidic conditions to acylated ortho-benzoyloxyphenylacetic acid esters. When the same reaction is carried out under neutral conditions, the reaction product is the 3-O-acyl-glucosyl-5-O-glucosyl-7-hydroxy coumarin.     

Nitrite-mediated decarboxylative conjugation of caffeic acid with glutathione under mildly acidic conditions

In acetate buffer, pH 4, at room temperature, nitrite ions can mediate an unusual decarboxylative conjugation of caffeic acid with glutathione leading to novel isomeric 2-(3,4-dihydroxyphenyl)-2-S-glutathionylacetaldehyde oximes. These results hint at a possible role of endogenous and/or dietary glutathione in the mechanisms by which caffeic acid can affect the burden of carcinogenic N-nitroso compounds in the digestive tract.     

Glycosaminoglycans and acidic glycoproteins in rabbit uterus under estrogenic conditions

Uterine slices obtained from the estrogen-treated rabbits were digested with pronase. Glycosaminoglycans and acidic glycopeptides were then isolated by Dowex 1 column chromatography and preparative electrophoresis on celulose acetate membrane (Separax), in succession.Each subfraction thus obtained was identified by the mobility on Separax electrophoresis and the digestibility with mucopolysaccharidases (Streptomyces hyaluronidase, testicular hyaluronidase, chondroitinase AC, chondroitinase ABC and heparinase). The resulting data showed that each complex saccharide (hyaluronic acid, heparan sulfate, chondroitin sulfate A, chondroitin sulfate C, dermatan sulfate, sulfated glycopeptide and sialoglycopeptide) was separated into 2–5 fractions, indicating charge and/or molecular heterogeneity of each complex saccharide.     

Equilibrium and kinetic folding of hen egg-white lysozyme under acidic conditions

The equilibrium and kinetic folding of hen egg-white lysozyme was studied by means of circular dichroism spectra in the far- and near-ultraviolet (UV) regions at 25 degrees C under the acidic pH conditions. In equilibrium condition at pH 2.2, hen lysozyme shows a single cooperative transition in the GdnCl-induced unfolding experiment. However, in the GdnCl-induced unfolding process at lower pH 0.9, a distinct intermediate state with molten globule characteristics was observed. The time-dependent unfolding and refolding of the protein were induced by concentration jumps of the denaturant and measured by using stopped-flow circular dichroism at pH 2.2. Immediately after the dilution of denaturant, the kinetics of refolding shows evidence of a major unresolved far-UV CD change during the dead time (<10 ms) of the stopped-flow experiment (burst phase). The observed refolding and unfolding curves were both fitted well to a single-exponential function, and the rate constants obtained in the far- and near-UV regions coincided with each other. The dependence on denaturant concentration of amplitudes of burst phase and both rate constants was modeled quantitatively by a sequential three-state mechanism, U<-->I<-->N, in which the burst-phase intermediate (I) in rapid equilibrium with the unfolded state (U) precedes the rate-determining formation of the native state (N). The role of folding intermediate state of hen lysozyme was discussed.     

Formation of a diazoquinone-type mutagen from acetaminophen treated with nitrite under acidic conditions

Acetaminophen (paracetamol) showed mutagenicity to Salmonella typhimurium TA100 and TA98 either with or without S9 mix when treated with excess nitrite in acidic solution. The mutagen was isolated and identified as 4-acetylamino-6-diazo-2,4-cyclohexadienone. Mutation tests and product analysis by high-performance liquid chromatography, however, suggested that this type of mutagen was hardly formed in the digestive tract of normal subjects.     

Arsonic acid-presenting superparamagnetic iron oxide for pH-responsive aggregation under slightly acidic conditions

We report the synthesis and characterization of the arsonic acid-presenting superparamagnetic iron oxide (SPIO). We used arsonoacetic acid as the ligand for SPIOs in aqueous media. The surface modification of the SPIOs was accomplished via the ligand exchange from undecanoic acid to the carboxyl moiety of arsonoacetic acid. Consequently, the well-dispersed arsonic acid-presenting SPIOs in water were obtained. We found that the dispersion state of the arsonic acid-presenting SPIOs can be sharply regulated by pH changes in the biological significant region. The well dispersion state of the arsonic acid-presenting SPIOs can be maintained at the neutral pH region. In contrast, the arsonic acid-presenting SPIOs can sensitively form the aggregation below pH 6.1. Moreover, these dispersion states can be controlled reversibly by the pH alteration in the narrow region.     

Reaction pathways of glucose oxidation by ozone under acidic conditions

The ozonation of d-glucose-1-¹³C, 2-¹³C, and 6-¹³C was carried out at pH 2.5 in a semi-batch reactor at room temperature. The products present in the liquid phase were analyzed by GC-MS, HPAEC-PAD, and ¹³C NMR spectroscopy. Common oxidation products of glucose have also been submitted to identical ozonation conditions. For the first time, a pentaric acid was identified and its formation quantitatively correlated to the loss of C-6 of glucose in the form of carbon dioxide. Potential mechanisms for the formation of this pentaric acid are discussed. The well-accepted pathway involving the anomeric position in glucose, gluconic acid, arabinose, and carbon dioxide is reinvestigated. The origin of small molecules such as tartaric, erythronic, and oxalic acids is clarified. Finally, new reaction pathways and tentative mechanisms consistent with the formation of ketoaldonic acids and smaller acids are proposed.     

Differential interactions of apo- and holocytochrome c with acidic membrane lipids in model systems and the implications for their import into mitochondria

Monomolecular layers of lipid extracts of microsomal, mitochondrial outer and inner membranes, and pure lipid species have been used to measure their interaction with apo- and holocytochrome c. Large differences were observed both with respect to the nature and the lipid specificity of the interaction. The initial electrostatic interaction of the hemefree precursor apocytochrome c with anionic phospholipids is followed by penetration of the protein in between the acyl chains. Apocytochrome c shows similar interactions for all anionic lipids tested. In strong contrast the holoprotein discriminates enormously between cardiolipin for which it has a high affinity and phosphatidylserine and phosphatidylinositol for which it has a much lower affinity. For these latter lipids the interaction with cytochrome c is primarily electrostatic. The cytochrome c-cardiolipin interaction shows several unique features which suggest the formation of a specific complex between the two molecules. These properties account for the preference in interaction of the apoprotein with the lipid extract of the outer mitochondrial membrane over that of the endoplasmic reticulum and the large preference of cytochrome c for the inner over that of the outer mitochondrial membrane lipid extract. Only apocytochrome c was able to induce close contacts between monolayers of the mitochondrial outer membrane lipids and vesicles of mitochondrial inner membrane lipids. Experiments with fragments of both protein and unfolding experiments with cytochrome c revealed that the differences in interaction between the two proteins are mainly due to differences in their tertiary structure and not the presence of the heme group itself. The initial unfolded structure of apocytochrome c is responsible for the high penetrative power of the protein and its ability to induce close membrane contact, whereas the folded structure of cytochrome c is responsible for the specific interaction with cardiolipin. The results are discussed in the light of the apocytochrome c import process in mitochondria and suggest that lipid-protein interactions contribute to targeting the precursor toward mitochondria and are important for its translocation across the outer mitochondrial membrane and the final localization of cytochrome c toward the outside of the inner mitochondrial membrane.     

Glycosaminoglycans and acidic glycoproteins in rabbit uterus under estrogenic conditions.

Uterine slices obtained from the estrogen-treated rabbits were digested with pronase. Glycosaminoglycans and acidic glycopeptides were then isolated by Dowex 1 column chromatography and preparative electrophoresis on cellulose acetate membrane (Separax), in succession. Each subfraction thus obtained was identified by the mobility on Separax electrophoresis and the digestibility with mucopolysaccharidases (Streptomyces hyaluronidase, testicular hyaluronidase, chondroitinase AC, chondroitinase ABC and heparinase). The resulting data showed that each complex saccharide (hyaluronic acid, heparan sulfate, chondroitin sulfate A, chondroitin sulfate C, dermatan sulfate, sulfated glycopeptide and sialoglycopeptide) was separated into 2-5 fractions, indicating charge and/or molecular heterogeneity of each complex saccharide.     

Solution of ribosomal proteins under mild conditions.

Ribosomal proteins from two eucaryotic species, prepared by either the guanidine . HCl or LiCl . urea method and subsequently dissolved in 8 M urea were found to be largely retained in solution after removal of the urea by dialysis against a solution of low ionic strength (0.05 M Tris . HCl, pH 7.6, 0.025 M KCl, 0.005 M magnesium acetate) and centrifugation at 100,000 times g. The protein composition of this preparation was virtually identical to that of the original urea-containing solution as determined by two-dimensional polyacrylamide gel electrophoresis. Thus, there exists a very simple method for obtaining the bulk of the ribosomal proteins in solution under conditions where ribosomes themselves are stable.     

Modelling interactions between phytoplankton and bacteria under nutrient-regenerating conditions

The interactions of phytoplankton and bacteria in a nitrogen-limitedsteady-state system with an organic nitrogen compound or ammoniumas the sole nitrogen source were modelled. The effects of variousalgal excretion rates and two different mathematical representationsof excretion were examined. The model predicted that higherexcretion elevated the bacterial steady-state biomass, and loweredthe algal biomass. Bacterial respiration, which directly determinednitrogen regeneration, had an important effect on the system.The bacterial growth yield in the model was mainly a functionof the growth rate, and not of the nitrogen:carbon ratio ofthe substrate. In one version of the model, where the excretionof organic carbon increased with decreasing growth rate, themodel started to oscillate when the multiplication product ofmaximum specific excretion of excreted organic carbon (EOC)and the bacterial yield on EOC exceeded the dilution rate, irrespectiveof the form of nitrogen (ammonium or dissolved organic nitrogen)in the medium. The model results were compared with chemostatexperiments with the alga Emiliania huxleyi and a bacterialisolate in pure and mixed culture at two different dilutionrates. The carbon and nitrogen biomass of the bacteria was {smalltilde}1.5 times higher in mixed culture than in pure culture.In the experiments with low dilution rate, the recovery of nitrogenin the form of biomass, ammonium or amino acids was low, suggestingthe excretion by the algae of a refractory nitrogen-containingproduct which the bacteria could not use.