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The ms2io6A37 modification of tRNA in Salmonella typhimurium regulates growth on citric acid cycle intermediates.

The modified nucleoside 2-methylthio-N-6-isopentenyl adenosine (ms2i6A) is present in position 37 (adjacent to and 3' of the anticodon) of tRNAs that read codons beginning with U except tRNA(i.v. Ser) in Escherichia coli. In Salmonella typhimurium, 2-methylthio-N-6-(cis-hydroxy)isopentenyl adenosine (ms2io6A; also referred to as 2-methylthio cis-ribozeatin) is found in tRNA, most likely in the species that have ms2i6A in E. coli. Mutants (miaE) of S. typhimurium in which ms2i6A hydroxylation is blocked are unable to grow aerobically on the dicarboxylic acids of the citric acid cycle. Such mutants have normal uptake of dicarboxylic acids and functional enzymes of the citric acid cycle and the aerobic respiratory chain. The ability of S. typhimurium to grow on succinate, fumarate, and malate is dependent on the state of modification in position 37 of those tRNAs normally having ms2io6A37 and is not due to a second cellular function of tRNA (ms2io6A37)hydroxylase, the miaE gene product. We suggest that S. typhimurium senses the hydroxylation status of the isopentenyl group of the tRNA and will grow on succinate, fumarate, or malate only if the isopentenyl group is hydroxylated.

Chromosome Mapping↗

Carob pod: a new substrate for citric acid production by Aspergillus niger.

The production of citric acid from carob pod extract by A. niger in surface fermentation was investigated. A maximum citric acid concentration (85.5 g/L), citric acid productivity (4.07 g/L/d), specific citric acid production rate (0.18 g/g/d), and specific sugar uptake rate (0.358 g/g/d) was achieved at an initial sugar concentration of 200 g/L, pH of 6.5, and a temperature of 30 degrees C. Other kinetic parameters, namely, citric acid yield, biomass yield, specific biomass production rate, and fermentation efficiency were maximum at pH 6.5, temperature 30 degrees C, and initial sugar concentration 100 g/L. The external addition of methanol into the carob pod extract at a concentration up to 4% (v/v) improved the production of citric acid.

Aspergillus niger↗

Patterns of diversity of citric acid cycle enzymes.

The citric acid cycle performs a dual role in cell metabolism, acting as a source of both 'energy' and biosynthetic starting materials. The widespread occurrence of the cycle throughout Nature is an excellent example of the unity of biochemistry, but closer examination reveals that there is considerable diversity in the citric acid cycle of different organisms with respect to metabolic role, molecular enzymology and mode of regulation. Two enzymes of the cycle--citrate synthase and succinate thiokinase--have been found to exhibit particularly striking patterns of diversity in structure and catalytic and regulatory function. Some of these patterns show a correlation with the taxonomic groupings of the organisms and with their physiological characteristics. Comparative enzyme studies have a contribution to make to an ultimate understanding of the cycle and its cellular operation, and there are substantial benefits to be gained from interactive studies on both prokaryotic and eukaryotic systems.

Animals↗

Characterization of the bisphosphonate recognition site on hydroxyapatite using radioligand binding techniques with [14C]citric acid.

The present studies characterize the binding of [14C]citric acid to synthetic hydroxyapatite (HA) crystals. [14C]Citric acid specifically bound to HA and was dependent upon the concentration of HA in the assay. The binding of [14C] citric acid to HA reached equilibrium within 20 min and remained stable for at least 90 min. Dissociation of bound [14C]citric acid was biphasic in nature since both rapid and more slowly reversible binding components were detected. Saturation experiments also indicated that [14C]citric acid labeled two recognition sites with different affinity (KdH = 42 nM and KdL = 24,000 nM) and density (BmaxH = 161 fmol/micrograms HA and BmaxL = 8.8 pmol/micrograms HA). Ligand competition experiments revealed that compounds that are known to readily bind bone (e.g., sodium pyrophosphate, methylene diphosphonic acid, etidronate) potently inhibited the binding of [14C]citric acid to HA, whereas compounds known to have poorer affinity for bone (e.g., oxalic acid and GABA) did not. Computer analysis of these inhibition curves revealed specific ligand interactions at two different affinity recognition sites. The present results indicate that [14C]citric acid binds discrete sites on synthetic HA in a fashion consistent with a specific labeling of the bisphosphonate recognition site. Analysis of the binding of [14C]citric acid to HA provides a useful method to further explore the structure activity relationships of novel compounds that have binding affinity for bone.

Binding Sites↗

The potentiation effect of citric acid in aureomycin in turkeys.

Citric acid was used to potentiate Aureomycin by administering both materials through the drinking water of turkeys. Treatment groups receiving citric acid consistently had higher blood levels of Aureomycin than corresponding treatments receiving Aureomycin alone. Highly significant differences in tetracycline blood levels were obtained at 24 hr in one experiment and at 48 hr in a second experiment favoring citric acid treated birds.

Administration, Oral↗

Fatty acid oxidation, substrate shuttles, and activity of the citric acid cycle in hepatocellular carcinomas of varying differentiation.

Fatty acid oxidation, reconstituted substrate shuttles, and the activity of the citric acid cycle were studied in mitochondria isolated from Becker transplantable hepatocellular carcinoma H-252 AND Host livers, and the results were compared with those obtained with Morris hepatomas 7288CTC and 5123C. Whereas the activities of the malate-aspartate and the alpha-glycerophosphate shuttles were only slightly lower than those of host livers, the activity of the fatty acid shuttle was much lower in H-252 mitochondria. Oxygen uptake and CO2 production associated with the oxidation of fatty acids was much lower in tumors H-252 and 7288CTC, compared with host livers, whereas tumor 5123C mitochondria show a high capacity to oxidize fatty acids. Ketogenesis and beta-hydroxybutyrate dehydrogenase activity were also lower in tumor H-252 mitochondria. However, neither oxygen uptake associated with the oxidation of other respiratory substrates nor CO2 production from succinate or malate was strikingly elevated in these tumors. These factors suggest that the respiratory phosphorylation chain and activity of the citric acid cycle are fully functional in tumors H-252 and 7288CTC. The defects responsbile for the lower rates of fatty acid oxidation in these tumors probably involves the beta-oxidation pathway, as well as the activation of fatty acids. The impairment of fatty acid oxidation may explain the lower activity of the reconstituted fatty acid shuttle for transporting reducing equivalents into H-252 mitochondria. The different properties with regard to fatty acid oxidation in Morris hepatoma 5123C, compared with those in Becker H-252- AND Morris hepatoma 7288CTC, may reflect the different extent of differentiation in these tumors, the former being a slow-growing, well-differentiated tumor, whereas the latter represent tumors that are less differentiated and of more rapid growth rate.

Animals↗

Morphological alterations associated with the cytotoxic and cytostatic effects of citric acid on cultured human dental pulp cells.

Citric acid exerts potential harmful effects on the pulp when used for root surface demineralization, smear layer removal, and dentin etching. Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, we found that incubation of cultured human dental pulp cells in medium containing 0.5% (pH 4.74) or 1.0% (pH 3.42) of citric acid for 2 h lead to 25% and 48% of cell death, respectively. Cytotoxicity of citric acid was associated with its acidity. Exposure of cells to pure 1% citric acid (pH 2.26) for 60 s lead to immediate cell death. Cytotoxicity was usually preceded by cell retraction, cell surface blebbing, and finally uptake of trypan blue, implicating the presence of cell membrane damage. A medium containing 0.05% citric acid can retard the growth of pulp cells. These results indicate that adequate protection of the pulp is important, especially when the remaining dentin is thin in deep carious lesions or in the presence of accessory canals on the root surface.

Acid Etching, Dental↗

Decalcification of root canal dentine by citric acid, EDTA and sodium citrate.

AIM: To measure the demineralization capability of 1 and 10% citric acid, 10% sodium citrate and 17% EDTA during immersions of 5, 10 and 15 min on root canal dentine. METHODOLOGY: Crowns were sectioned from eight maxillary canines. The cementum was removed from the cervical third of the roots to expose the dentine. Canals were prepared using a handpiece-mounted Largo Peeso reamer. A 3-mm thick cross-sectional slice was obtained from the cervical third of each root. Each slice was sectioned into four equal parts. These specimens were assigned to one of four groups (n = 8) for the application of 1% citric acid, 10% citric acid, 10% sodium citrate or 17% EDTA. Each specimen underwent three successive 5-min immersions in each solution at room temperature. The solutions were not renewed between immersions. Two millimetres of solution were collected from the extracts and lanthanum oxide was added for the calcium reading by spectrophotometry. To compare the amounts of calcium removed by each solution, the Friedman test was used for the global comparison and the Wilcoxon test for paired comparisons. Differences between groups were evaluated using the Kruskal-Wallis test for the global comparison and Mann-Whitney test for paired comparisons. RESULTS: Overall, 1 and 10% citric acid were more effective than EDTA or sodium citrate at the three immersion times (P < 0.001); 10% citric acid was more effective than 1% citric acid (P < 0.001). EDTA and 1 and 10% citric acid showed decreasing effectiveness with time, and the decrease was significant for citric acid at both concentrations (P < 0.001). Although sodium citrate removed little calcium during the three time periods, the small increase recorded was significant (P < 0.01). CONCLUSIONS: Citric acid at 10% was the most effective decalcifying agent, followed by 1% citric acid, 17% EDTA and 10% sodium citrate.

Calcium↗

Pulmonary and cardiovascular changes in hyperreactive rats from citric acid aerosols.

Administration of aerosols of citric acid to anesthetized spontaneously breathing hyperreactive rats produced reversible increases in respiratory rate (f) and pleural pressure (Ppl) accompanied by hypotension and bradycardia. In contrast, Fischer rats, which do not have bronchial hyperreactivity, failed to respond to citric acid aerosols. The effects of various treatments on citric acid-induced changes in f, Ppl and blood pressure were studied. Vagotomy, cromolyn sodium (1 and 10 mg/kg iv), mecamylamine, (2 mg/kg iv), FPL-55712 (3 mg/kg iv), and BW 755C (10 mg/kg iv) inhibited markedly the responses to citric acid, whereas atropine (2 mg/kg iv) produced weak inhibition. Methysergide (1 mg/kg iv), indomethacin (1 mg/kg iv), and a prostanoid antagonist, L-640,035, (5 mg/kg iv) were completely ineffective. The results suggest that citric acid-induced bronchoconstriction in hyperreactive rats may be reflex mediated and that leukotrienes may be involved in the response.

Aerosols↗

Effects of citric acid treatment on autogenous free graft coverage of localized recession.

Citric acid root conditioning prior to free gingival autograft (FGA) coverage of denuded roots may increase the likelihood of reattachment, but controlled studies to determine the value of the procedure have not been done on human subjects. This investigation was done to compare the effects of FGA root coverage with and without citric acid pretreatment. Nineteen patients with 30 affected teeth participated in the study. Fifteen teeth received FGAs without citric acid pretreatment and 15 teeth received FGAs with citric acid pretreatment. Measurements of gingival recession and sulcus depth were made preoperatively and 30, 60 and 90 days following the surgical procedure. Free gingival autografts with and without citric acid pretreatment each significantly reduced the amount of exposed root, but no differences between groups were found. Except for the 90-day single tooth control group, mean sulcus depths were significantly decreased at 30, 60 and 90 days, but there were no differences noted between single tooth citric acid and control groups or multiple tooth citric acid and control groups. Citric acid treatment of affected roots prior to FGA coverage does not appear to be justifiable.

Adult↗

Partial purification of a growth factor from orange juice which affects citrus tissue culture and its replacement by citric Acid.

Orange (Citrus sinensis L. Osbeck) tissue cultures required a supply of orange juice to the medium for their vigorous growth. The growth-promoting activity of juice seemed to involve both cell division and cell enlargement. Juice had no promotive activity in bioassays for auxins, gibberellins, and cytokinins. The growth promoting activity of juice was mostly transferred into 1-butanol upon partition at pH 2. Gas chromatographic analysis of this acid 1-butanol fraction revealed large amounts of citric acid and negligible amounts of other organic acids. Supply of pure citric acid to the medium, alone or in combination with different concentrations of juice, indicated that citric acid replaces most of the requirement for juice.It seems that citric acid, which is a natural component of citrus juice, is responsible for the major part of the growth-promoting activity of the juice. The significance of citric acid as a growth factor in tissue cultures and the reasons for the dependence of citrus tissue cultures on external supply of citric acid are discussed.

Journal Article↗

Effects of citric acid treatment on pedicle flap coverage of localized recession.

Citric acid conditioning of denuded roots prior to laterally positioned pedicle flap (LPPF) coverage may increase the likelihood of reattachment, but no controlled studies to determine the efficacy of the procedure in managing naturally occurring defects in humans have been reported. This investigation compared the effects of LPPF root coverage with and without citric acid (pH1) pretreatment in a group of patients with gingival recession and associated denuded roots. Thirty-six patients participated in the study. Eighteen received LPPFs without citric acid pretreatment and 18 received LPPFs with citric acid pretreatment. Gingival recession and sulcus depth were measured preoperatively and 30, 60 and 90 days after the surgical procedure. LPPFs both with and without citric acid pretreatment reduced significantly the amount of exposed root, but no differences between groups were found. For sulcus depth, the 90-day value for the citric acid group was significantly increased over the preoperative level, but no other changes were noted. There appears to be no clinical justification for using citric acid conditioning as an adjunct to LPPF coverage of denuded roots.

Adult↗

Influence of medium components and metabolic inhibitors on citric acid production by Penicillium simplicissimum.

Penicillium simplicissimum excreted more than 100 mmol citric acid l-1 [2.9 mmol (g dry wt)-1; 9 d] if an industrial filter dust (> 50% ZnO) providing a high extracellular buffering capacity was present in the medium. A similar specific [2 mmol (g dry wt)-1], but lower absolute (26 mmol l-1), citric acid excretion occurred in the absence of an extracellular buffer and if amino acids or urea were used as nitrogen source. P. simplicissimum excreted no citric acid under conditions where Aspergillus niger produces citric acid (deficiency of trace elements, low pH and reduced biomass formation). Citric acid excretion by P. simplicissimum always paralleled biomass formation and occurred in a pH range between 4 and 7. This indicated that different imbalances of metabolism were responsible for citric acid excretion in A. niger and P. simplicissimum. However, provided a high extracellular buffering capacity was present, the response of the Penicillium system to different carbon and nitrogen sources was similar to the Aspergillus system. In contrast, the metals iron and copper had virtually no effect on citric acid excretion compared with A. niger. Estimation of intracellular citric acid, as well as the effects of the uncoupler 2,4-dinitrophenol, and the H(+)-ATPase inhibitor sodium orthovanadate, led to the conclusion that the buffer-stimulated citric acid efflux was dependent on metabolic energy and an energized plasma membrane, respectively. Despite similarities to the Aspergillus system, a different mechanism for buffer-stimulated citric acid excretion by P. simplicissimum seems probable.

Antimetabolites↗

The fate of Salmonella enteritidis PT4 in home-made mayonnaise prepared with citric acid.

The fate of Salmonella enteritidis PT4 in home-made mayonnaise prepared with citric acid solution (citric acid concentration of > or = 4.98% (w/v)) was investigated. It was found that pH of mayonnaise is closely related to the ratio of egg yolk to citric acid, and the inactivation rate of the micro-organisms increases as the ratio decreases and/or incubation temperature increases. To achieve Salm. enteritidis PT4-free home-made mayonnaise prepared with pure lemon juice (citric acid concentration > or = 5% (w/v)), it is recommended that the pH should be 3.30 or below, or, in practice, at least 20 ml pure lemon juice per fresh egg yolk should be used. For the use of 20-35 ml pure lemon juice per egg yolk, the product should be held at 22 degrees C or over for at least 72 h and for the use of over 35 ml pure lemon juice per egg yolk, for at least 48 h before consumption or refrigeration.

Citric Acid↗

Modeling the adsorption of Cd(II) onto goethite in the presence of citric acid.

The adsorption of cadmium onto goethite in the presence of citric acid was measured as a function of pH and cadmium concentration at 25 degrees C. Potentiometric titrations were also performed on the system. Cadmium adsorption onto goethite was enhanced above pH 4 in the presence of 50 microM, 100 microM and 1 mM citric acid. While there was little difference between the enhancements caused by 50 and 100 microM citric acid below pH 6, above pH 6 further enhancement is seen in the presence of 100 microM citric acid. When 1 mM citric acid was present, the enhancement of cadmium adsorption was greater below pH 6, with increased Cd(II) adsorption down to pH 3.5. However, above pH 6, 1 mM of citric acid caused slightly less enhancement than the lower citric acid concentrations. ATR-FTIR spectra of soluble and adsorbed citrate-cadmium species were measured as a function of pH. At pH 4.6 there was very little difference between the ternary Cd(II)-citric acid-goethite spectrum and the binary citric acid-goethite spectrum. However, spectra of the ternary system at pH 7.0 and 8.7 indicated the presence of additional surface species. Further analysis of the spectra suggested that these were metal-ligand outer-sphere complexes. Data from the adsorption experiments and potentiometric titrations of the ternary Cd(II)-citric acid-goethite system were fitted by an extended constant-capacitance surface complexation model. The spectroscopic data were used to inform the choice of surface species. Three reactions in addition to those for the binary Cd(II)-goethite and citric acid-goethite systems were required to describe all of the data. They were [formula in text], [formula in text], and [formula in text]. Neither the spectroscopy nor the modeling suggested the formation of a ternary inner-sphere complex or a surface precipitate under the conditions used in this study.

Journal Article↗