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On the origin of the non-haemic iron transferrin ESR signal: ESR investigations on histidine-iron-citric acid systems.

The nature of the high spin ferric iron complex located at g = 4.3 has been investigated by means of electron spin resonance spectroscopy and polarography. It could be shown that two complexes each exist in the acid and alkaline pH region, and that the iron is bound to two histidines, three citric acids, and probably to one bicarbonate. These results agree well with previous findings according to which the ligand field of iron should be composed mainly of oxygen and nitrogen atoms. Another low-field signal located at g = 3.6 appears in the pH range from 2 to 7 only and exhibits its maximum where the g = 4.3 signal has its minimum. Its exact nature is still unknown but it seems to represent some intermediate state of the ternary Fe3+-histidine-citric acid complex. When citric acid is used, the spin concentration seems to be always larger than in the case of ascorbic acid. Since the effect obtained with ascorbic acid and citric acid seems to be similar, it may be concluded that the biological function of both of the acids might be somehow related to each other.

Ascorbic Acid↗

Dentin demineralization. The effects of citric acid concentration and application time.

Preliminary work has shown that the rate of dentin demineralization increases with increasing concentrations of citric acid. This rate subsequently diminishes at much higher concentrations. The purpose of this study was to more precisely identify the citric acid concentration which produces peak dentin demineralization and to determine if this demineralization process is time dependent. Flat dentin surfaces were prepared on the buccal and lingual sides of 15 bovine molars. 8 depressions were made in each dentin surface using a #8 round bur in a high-speed handpiece with air-water coolant. Various concentrations of citric acid solutions (weight per cent) were prepared, e.g., 0%, 10%, 20%, 25%, 30%, 35%, 40%, 65% and their respective pH's recorded. 3 microliters of each citric acid solution were placed in individual depressions on the dentin surfaces and left undisturbed for 1, 2 or 3 min. Cotton pellets were used to soak up the citric acid solution, along with any dissolved calcium, and were subsequently placed in 10 ml of 18 Me omega water. The parts per million calcium found in each water sample were determined using atomic absorption spectrophotometry. Peak dentin demineralization for 1-, 2- and 3-min application times occurred at 30% (pH = 1.55), 25% (pH = 1.62) and 25% (pH = 1.62) citric acid concentrations/(pH), respectively. Dentin demineralization was found to be time-dependent for all citric acid solution concentrations. The clinical significance of these findings is discussed.

Analysis of Variance↗

Global analysis of gene expression during development and ripening of citrus fruit flesh. A proposed mechanism for citric Acid utilization.

Microarrays of cDNA have been used to examine expression changes of 7000 genes during development and ripening of the fruit flesh of self-incompatible Citrus clementina, a non-climateric species. The data indicated that 2243 putative unigenes showed significant expression changes. Functional classification revealed that genes encoding for regulatory proteins were significantly overrepresented in the up-regulated gene clusters. The transcriptomic study together with the analyses of selected metabolites highlighted key physiological processes occurring during citrus fruit development and ripening such as water accumulation, carbohydrate build-up, acid reduction, pigment substitutions (carotenoid accumulation and chlorophyll decreases) and ascorbic acid diminution. Often, the combined analyses strongly suggested prevalence of specific metabolic alternatives. This observation has been exemplified with the proposal for a mechanism for citrate utilization, a process of much importance in citrus industry. Microarray data validated by real-time RT-PCR suggested that citrate was sequentially metabolyzed to isocitrate, 2-oxoglutarate and glutamate. Thereafter, glutamate was both utilized for glutamine production and catabolyzed through the gamma-aminobutirate (GABA) shunt (GABA --> succinate semialdehyde --> succinate). This last observation appears to be of special relevance since it links the proton consuming reaction glutamate + H(+)--> GABA + CO(2) with high acid levels. GG-MS determinations showed that glutamate was constant while GABA levels decreased at ripening in agreement with a feasible activation of the GABA shunt during acid catabolism. This suggestion provides a convincing explanation for the strong reduction of both citrate and cytoplasmatic acidity that takes place in citrus fruit flesh during development and ripening.

Amino Acids↗

Root dentin morphology after different modes of citric acid and tetracycline hydrochloride conditioning.

The purpose of this study was to assess the effect of citric acid and tetracycline HCl application to dentin surfaces by a "passive dripping" or an "active burnishing" technique. Twenty dentin blocks were prepared from freshly extracted non-diseased human impacted third molars. The blocks were root planed and randomly assigned to two groups for treatment with either citric acid or tetracycline HCl. The duration of treatment was 30, 60, 120, or 240 seconds. Control blocks were treated with distilled water. After treatment the blocks were processed for observation and measurements in the scanning electron microscope (SEM). Application of either of the acid solutions resulted in removal of the smear layer. Measurements indicated a time dependent increase in the mean dentinal tubule orifice diameter ranging from 1.05 microns in control specimens to 3.18 microns after 4 minutes treatment (citric acid group). The increase in tubule diameter was significantly greater (P less than or equal to 0.01) for both citric acid treatment modalities than tetracycline HCl treatment. There was also a time dependent increase in the depth of penetration as measured by a trumpeting of the tubule profiles, and this penetration was significantly greater (P less than or equal to 0.01) after citric acid treatments. Passive or active application of the acids did not seem to have any major impact on the measurements or on the surface morphology. It was concluded that citric acid causes more extensive changes than tetracycline HCl and that the mode of application of the agent is probably not critical.

Acid Etching, Dental↗

Citric acid production by selected mutants of Aspergillus niger from cane molasses.

The present investigation deals with citric acid production by some selected mutant strains of Aspergillus niger from cane molasses in 250 ml Erlenmeyer flasks. For this purpose, a conidial suspension of A. niger GCB-75, which produced 31.1 g/l citric acid from 15% (w/v) molasses sugar, was subjected to UV-induced mutagenesis. Among the 3 variants, GCM-45 was found to be a better producer of citric acid (50.0 +/- 2a) and it was further improved by chemical mutagenesis using N-methyl, N-nitro-N-nitroso-guanidine (MNNG). Out of 3,2-deoxy-D-glucose resistant variants, GCMC-7 was selected as the best mutant, which produced 96.1 +/- 1.5 g/l citric acid 168 h after fermentation of potassium ferrocyanide and H2SO4 pre-treated blackstrap molasses in Vogel's medium. On the basis of kinetic parameters such as volumetric substrate uptake rate (Qs), and specific substrate uptake rate (qs), the volumetric productivity, theoretical yield and specific product formation rate, it was observed that the mutants were faster growing organisms and produced more citric acid. The mutant GCMC-7 has greater commercial potential than the parental strain with regard to citrate synthase activity. The addition of 2.0 x 10(-5) M MgSO4 x 5H2O into the fermentation medium reduced the Fe2+ ion concentration by counter-acting its deleterious effect on mycelial growth. The magnesium ions also induced a loose-pelleted form of growth (0.6 mm, diameter), reduced the biomass concentration (12.5 g/l) and increased the volumetric productivity of citric acid monohydrate (113.6 +/- 5 g/l).

Aspergillus niger↗

Detection of exogenous citric acid in fruit juices by stable isotope ratio analysis.

A new method has been developed for measuring the D/H ratio of the nonexchangeable sites of citric acid by isotope ratio mass spectrometry (IRMS). Pure citric acid is transformed into its calcium salt and subsequently analyzed by pyrolysis-IRMS. The citric acid isolated from authentic fruit juices (citrus, pineapple, and red fruits) systematically shows higher D/H values than its nonfruit counterpart produced by fermentation of various sugar sources. The discrimination obtained with this simplified method is similar to that obtained previously by applying site specific isotopic fractionation-nuclear magnetic resonance (SNIF-NMR) to an ester derivative of citric acid. The combination of carbon 13 and deuterium measurements of extracted citric acid is proposed as a routine method for an optimum detection of exogenous citric acid in all kinds of fruit juices.

Beverages↗

The effect of Chlamydia trachomatis infection of the prostate gland on the concentration of citric acid.

INTRODUCTION: The objective of the study was to assess the relationship between Chlamydia trachomatis (C.t.) infection of the prostate and the concentration of citric acid. MATERIALS AND METHODS: The study involved 60 patients with chronic prostatitis (NIH III). Urethral swabs and expressed prostatic secretions (EPS) were collected for analysis. The urethral swabs were tested for PMNs and the presence of Neisseria gonorrhoeae and C.t., while the EPS were analyzed to determine PMN count, C.t., and citric acid concentration. The DFA or LCR method was used for C.t. diagnosis. The concentration of citric acid was measured using the UV method. RESULTS: Inflammation of the prostate (PMNs >/=10/field) was diagnosed in 58.3 of the patients. C.t. infection was found in 20%, including 8.3% with only the urethra affected and 10% with only the prostate. One patient had both the urethra and the prostate infected. A reduction in the concentration of citric acid in EPS was observed in 56.7% of the men. In 88.2% of the patients, reduced citric acid concentration was accompanied by an elevated PMN count in the EPS. All patients with C.t. infection of the prostate showed a reduced concentration of citric acid. In five patients with urethral infection, lack of a decrease in this parameter was noted in one. In all the patients with chlamydial infection, irrespective of localization, a high PMN count was observed in the EPS. CONCLUSIONS: Determination of the concentration of citric acid in the prostatic fluid is a good indicator of prostatitis. C.t. infection of the prostate gland is accompanied by a decrease in the concentration of citric acid.

Adolescent↗

Effect of mineral supplements to citric acid on enamel erosion.

The aim of this study was to evaluate the effect of mineral supplements to citric acid (1%; pH 2.21) on enamel erosion under controlled conditions in an artificial mouth. From each of 156 bovine incisors one polished enamel sample was prepared. The samples were divided among 13 experimental groups (n=12). In group 1 citric acid only was used (control). In groups 2-10 either calcium, phosphate or fluoride in various low concentrations was admixed to the citric acid. In groups 11-13 the citric acid was supplemented with a mixture of calcium, phosphate and fluoride. For demineralisation the specimens were rinsed with the respective solution for 1 min, immediately followed by a remineralisation period with artificial saliva (1 min). The specimens were cycled through this alternating procedure five times followed by rinsing for 8 h with artificial saliva. The de- and remineralisation cycle was repeated three times for each specimen interrupted by the 8 h-remineralisation periods. Before and after the experiments, the specimens were examined using microhardness testing (Knoop hardness) and laser profilometry. Hardness loss and enamel dissolution was significantly higher for the controls as compared to the remaining groups. Significantly lowest hardness loss for all groups was recorded for group 12 with admixture of calcium, phosphate and fluoride to citric acid. The significantly highest enamel loss was recorded for the controls compared to all other samples. Groups 3 and 4 revealed significantly lower and higher tissue loss compared to the remaining groups (2-13), respectively. The other groups did not differ significantly from each other. Modification of citric acid with calcium, phosphate and fluoride exerts a significant protective potential with respect to dental erosion. However, with the low concentrations applied enamel dissolution could not be completely prevented.

Animals↗

Citric acid fermentation of sugars purified with chelating resin.

Noguchi, Yuichi (University of Wisconsin, Madison), and Marvin J. Johnson. Citric acid fermentation of sugars purified with chelating resin. J. Bacteriol. 82:538-541. 1961.-A new, independently isolated strain of Aspergillus niger capable of giving high yields of citric acid in submerged culture was found to show the same behavior toward iron, zinc, and manganese as a previously studied strain. Citric acid accumulation did not occur in the presence of manganese. Best citric acid production was obtained in the presence of limited amounts of iron and zinc. Use of a chelating ion exchange resin was found to be an excellent method of removing polyvalent metals from sugars, either for analytical purposes or for fermentation. Commercial glucose, after resin treatment, gave citric acid yields of more than 80% when supplemented with iron and zinc. Unpurified glucose was converted to citric acid in good yields by mycelium grown on resin-treated glucose.

Aspergillus↗

Oxygen requirements for growth and citric acid production of Yarrowia lipolytica.

During continuous cultivation of Yarrowia lipolytica N 1, oxygen requirements for growth and citric acid synthesis were found to depend on the iron concentration in the medium. A coupled effect of oxygen and iron concentrations on the functioning of the mitochondrial electron transport chain in Y. lipolytica N 1 was established. Based on the results obtained in continuous culture, conditions for citric acid production in a batch culture of Y. lipolytica N 1 were proposed. At relatively low pO(2) value and a high iron concentration, citric acid accumulation was as high as 120 g l(-1); the specific rate of citric acid synthesis reached 120 mg citric acid (g cells h)(-1). The mass yield coefficient was 0.87 and the energy yield coefficient was 0.31.

Aconitate Hydratase↗

Role of different additives and metallic micro minerals on the enhanced citric acid production by Aspergillus niger MNNG-115 using different carbohydrate materials.

The present investigation deals with the promotry effect of different additives and metallic micro minerals on citric acid production by Aspergillus niger MNNG-115 using different carbohydrate materials. For this, sugar cane bagasse was fortified with sucrose salt medium. Ethanol and coconut oil at 3.0% (v/w) level increased citric acid productivity. Fluoroacetate at a concentration of 1.0 mg/ml bagasse enhanced the yield of citric acid significantly. However, the addition of ethanol and fluoroacetate after 6 h of growth gave the maximum conversion of available sugar to citric acid. In another study, influence of some metallic micro-minerals viz. copper sulphate, molybdenum sulphate, zinc sulphate and cobalt sulphate on microbial synthesis of citric acid using molasses medium was also carried out. It was found that copper sulphate and molybdenum sulphate remarkably enhanced the production of citric acid while zinc sulphate was not so effective. However, cobalt sulphate was the least effective for microbial biosynthesis of citric acid under the same experimental conditions. In case of CuSO(4), the strain of Aspergillus niger MNNG-115 showed enhanced citric productivity with experimental (9.80%) over the control (7.54%). In addition, the specific productivity of the culture at 30 ppm CuSO(4) (Q(p) = 0.012a g/g cells/h) was several folds higher than other all other concentrations. All kinetic parameters including yield coefficients and volumetric rates revealed the hyper productivity of citric acid by CuSO(4) using blackstrap molasses as the basal carbon source.

Alcohols↗

Citraturic response to oral citric acid load.

It is possible that some orally administered citrate may appear in urine by escaping oxidation in vivo. To determine whether this mechanism contributes to the citraturic response to potassium citrate, we measured serum and urinary citrate for 4 hours after a single oral load of citric acid (40 mEq.) in 6 normal subjects. Since citric acid does not alter acid-base balance, the effect of absorbed citrate could be isolated from that of alkali load. Serum citrate concentration increased significantly (p less than 0.05) 30 minutes after a single oral dose of citric acid and remained significantly elevated for 3 hours after citric acid load. Commensurate with this change, urinary citrate excretion peaked at 2 hours and gradually decreased during the next 2 hours after citric acid load. In contrast, serum and urinary citrate remained unaltered following the control load (no drug). Differences of the citratemic and citraturic effects between phases were significant (p less than 0.05) at 2 and 3 hours. Urinary pH, carbon dioxide pressure, bicarbonate, total carbon dioxide and ammonium did not change at any time after citric acid load, and did not differ between the 2 phases. No significant difference was noted in serum electrolytes, arterialized venous pH and carbon dioxide pressure at any time after citric acid load and between the 2 phases. Thus, the citraturic and citratemic effects of oral citric acid are largely accountable by provision of absorbed citrate, which has escaped in vivo degradation.

Administration, Oral↗

Partitioning the respiratory effects of airway citric acid and normal saline in lambs.

Aspiration is common in the intubated human neonate. Thus, the ventilatory and blood gas responses to citric acid and saline instillation into different airway sites were studied in ten awake, unanesthetised lambs, breathing spontaneously via a tracheostomy tube. With a system of balloons, 1 ml of saline or citric acid was placed selectively into the midtrachea, the laryngeal area, or the lower trachea (lower tr). Changes in minute ventilation (VE), after a 30 sec baseline period, were measured 30 sec and 1 and 2 min after the challenge. Arterial blood gas changes were measured at 30 sec and 2 min. Major increases in VE were seen only when saline or citric acid was instilled into the lower tr, the citric acid responses exceeding saline ones. The arterial oxygen tension (PaO2) fell after lower tr saline, whereas the arterial CO2 tension (PaCO2) fell with midtracheal saline instillation. A rise in pH and a fall in PaCO2 accompanied citric acid given into the lower tr. An initial rise in PaO2 after citric acid into the lower tr was followed by a return to baseline despite hyperventilation. The ventilatory and blood gas changes with saline and citric acid depend on the site of airway instillation.

Airway Resistance↗

Citric acid aerosol as a potential smoking cessation aid.

We tested the ability of a citric acid aerosol to simulate the tracheal sensations produced by cigarette smoke and to satisfy smokers' desire for cigarettes. Fifteen smokers rated puffs from their own brand of cigarette, citric acid aerosol, a low tar and nicotine cigarette, and air. To focus on tracheal perceptions and pharmacologic effects of nicotine, we equated visual, olfactory and taste cues across conditions. Subjects rated the citric acid aerosol more similar to their own brand, more desirable, and more satisfying (after a block of puffs) than control puffs of air. It was also rated equal to or better than the low tar and nicotine cigarette. Subjects' own brands were rated best, although puffs of citric acid aerosol were of comparable strength and harshness. The results suggest that a nebulizer delivering citric acid in a fine mist might reduce craving for cigarettes in smokers attempting to quit and may thereby increase cessation rates.

Adult↗