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Stimulation by citric acid of calcium and phosphorus bioavailability in rats fed a calcium-rich diet.

The effect of dietary citric acid supplementation on calcium (Ca) and phosphorus (P) bioavailability was studied in rats fed 3 different diets containing 0.1, 0.5 or 1.0 g/100g Ca during 7 weeks. Citric acid supplementation increased intestinal Ca and P absorption and the Ca and P retention/intake ratio only in rats fed the 1% Ca diet. It also increased the P concentration of bone ashes in rats fed the 0.5% Ca diet (18.9 +/- 0.2 vs 17.5 +/- 0.5%) and the 1% Ca diet (20 +/- 0.1 vs. 19 +/- 0.3%), and the Ca bone ash concentration in rats fed the 1% Ca diet (36.7 +/- 0.4 vs. 35.7 +/- 0.5%). In rats fed the 1% Ca diet, plasma P concentration was decreased by citric acid supplementation (2.09 +/- 0.10 vs. 2.45 +/- 0.08 mmol/l) while urinary P excretion was increased (18.2 +/- 2.3 vs. 2.0 +/- 0.3 mmol/4 days), together with a decrease in plasma calcitriol concentration (54.4 +/- 2.6 vs. 79.6 +/- 2.5 ng/l), but no change of the circulating parathyroid hormone level. This study indicates that citric acid supplementation together with a Ca-rich diet allows to obtain an increased retention of Ca and P in bone. The prolonged administration of Ca citrate supplements may therefore help to increase bone mineral concentration.

Animals↗

Inhibition of fungal NADP+-isocitrate dehydrogenase by citric acid.

The variations observed during earlier studies in the activity of NADP+-isocitrate dehydrogenase (EC. 1.1.1.42) in a strain of Aspergillus niger were found to be related to the extent of washing of mycelium. As a result the mycelium washed four times with phosphate buffer (0.05 M, pH 7.5), the enzyme activity present in 4 and 8 days old fungal mycelia increased five- and two-fold, respectively. In vivo studies showed a complete loss of enzyme activity in mycelia resuspended in HCl-KCl buffer (0.02 M, pH 2.2) containing citric acid (13 mM or more). The in vitro studies revealed 50% loss of enzyme activity in presence of 3.6 to 5.2 mM citric acid. However, in case of Aspergillus niger ATCC 1015, which produced less citric acid than the above strain, a much higher citric acid concentration (13 to 26 mM) was required to cause 50% loss of enzyme activity. These findings suggest a correlation between citric acid inhibition of NADP+-isocitrate dehydrogenase and the ability of A. niger to accumulate citric acid in the medium.

Aspergillus niger↗

Citric acid demineralization of cementum and dentin: the effect of application pressure.

Previous work has shown that a 25-30% citric acid solution was the most effective concentration with which to demineralize dentin. The purpose of this investigation was to study the topography, using a scanning electron microscope, of root surfaces treated with a 30% citric acid solution using various application pressures. 20 freshly extracted human teeth were collected and stored in physiologic saline at room temperature. 3 root specimens, approximately 3x5x5 mm in size, were prepared from the coronal periodontally healthy area of each tooth. 30 specimens were root planed to expose dentin (dentin group) while the remaining 30 specimens were lightly scaled to remove periodontal soft tissue (cementum group). A freshly made 30% citric acid (CA) solution, (pH = 1.60), was applied to each of the experimental areas. Cotton pellets soaked in the citric acid solution were either placed (CAP), lightly rubbed (CAR) or heavily burnished (CAB) on the prepared root surface for 3 min. Pellets were resoaked every 30 s. The root sections were rinsed, fixed in glutaraldehyde, dehydrated in graded ethanol, critically point dried in liquid CO2 and sputter coated in gold. The treated surfaces were assessed for fibril tufting using scanning electron microscopy. Assessment was made of: (i) the % of surface area tufted; (ii) tufting depth (0-3); (iii) tufting density (1-3). Results of the study showed significantly more tufted surface area and greater tufting depth on both dentin and cementum for CAR and CAB compared to CAP. CAP produced a flat/mat fibril surface with no evidence of tufting.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Etching, Dental↗

Citric acid airway constriction in dogs with hyperreactive airways.

To understand the mechanisms underlying the bronchoconstrictor response to 10% citric acid administered for 5 min in Basenji-Greyhound (BG) dogs, we evaluated the protection afforded by atropine (0.2 and 0.4 mg/kg iv) and by aerosols of isoproterenol (1 mg/ml) and cromolyn sodium (20 mg/ml). In untreated dogs, citric acid increased pulmonary resistance by 4.6- to 11.5-fold and decreased dynamic compliance (Cdyn) to 45-55% of the control response. Isoproterenol and cromolyn sodium significantly reduced the response, whereas atropine did not. Moreover we have demonstrated in the arterial plasma of these dogs a slow-reacting substance (SRS) after, but not before, citric acid challenge. This SRS exhibits both pharmacologic properties and chemical characteristics similar to leukotrienes. We conclude that mediators of immediate-type hypersensitivity rather than reflex mechanisms play a dominant role in the production of airway constriction during citric acid (5-min) challenge in BG dogs.

Airway Resistance↗

Sensory properties of citric acid: psychophysical evidence for sensitization, self-desensitization, cross-desensitization and cross-stimulus-induced recovery following capsaicin.

In a first experiment, human subjects used a bipolar scale to rate the irritant sensation elicited by 10 sequentially repeated applications of either 3 ppm capsaicin or 250 mM citric acid on one side of the dorsal surface of the tongue, at 1 min intervals (30 s inter-stimulus interval). Citric acid-evoked irritation significantly increased across trials, consistent with sensitization. With capsaicin there was a large degree of inter- and intra-individual variation in successive ratings with no overall sensitization. Following the sequential stimulation series and a 10 min rest period, self- and cross-desensitization effects were tested in a two-alternative forced choice (2-AFC) paradigm by placing either citric acid or capsaicin on both sides of the tongue and asking subjects to indicate which side of the tongue yielded a stronger irritant sensation. Subjects also gave separate intensity ratings for irritation on each side of the tongue. Capsaicin self-desensitization was confirmed, while cross-desensitization to citric acid was not observed. In addition, citric acid self-desensitization and cross-desensitization to capsaicin were observed. In a second experiment a stronger capsaicin solution (33 ppm) was applied to one side of the tongue using cotton swabs. After the burning sensation elicited by capsaicin had disappeared, citric acid was applied bilaterally and cross-desensitization was observed using the same 2-AFC and rating procedures. This was followed by repeated re-application of citric acid at 1 min intervals to the capsaicin-treated side. The irritant sensation elicited by citric acid increased significantly, indicating a 'cross-stimulus-induced recovery' from capsaicin desensitization. In a final experiment we investigated the effect of the sodium channel blocker amiloride on the perceived irritation elicited by citric acid or capsaicin. Following application of amiloride to one side of the tongue with cotton swabs, either citric acid or capsaicin was applied bilaterally and subjects asked to perform a 2-AFC and intensity ratings. Amiloride significantly, albeit weakly, reduced the irritation elicited by citric acid while it weakly but significantly enhanced capsaicin-evoked irritation. These findings are discussed in terms of involvement of vanilloid and acid-sensitive ion channels in acid-evoked irritation and pain.

Adolescent↗

Optimal citric acid concentration for dentinal demineralization.

A number of acids have been shown to have a peak concentration above which the rate of enamel demineralization diminishes. Citric acid, used to demineralize dentin in periodontal regenerative procedures, is one such acid. The purpose of this study was to ascertain if there were an optimal concentration of citric acid with which to demineralize dentin. Various concentrations of citric acid were applied to bovine dentin and then examined with atomic absorption spectrophotometry to measure the amount of calcium dissolved in each concentration. The ppm calcium/% citric acid solution rose from 0.38 +/- 0.38 ppm/0% (control) to a peak of 7.54 +/- 2.26 ppm/24%, then fell to 2.43 +/- 0.59 ppm/80%, suggesting that there may be a peak concentration beyond which effective dentinal demineralization diminishes. The peak solution was pH = 1.42.

Animals↗

The use of citric acid to prolong the in vivo gastro-retention of a floating dosage form in the fasted state.

Gastro-retentive dosage forms have the potential to improve local therapy and decrease the variation in bioavailability that is observed with a number of commercially available immediate and modified release preparations. In this study, a dosage form has been developed, utilising freeze-dried calcium alginate beads, designed to float on the surface of the stomach contents thus prolonging the retention time. The aim of the study was to also assess the in vivo behaviour of the radio-labelled calcium alginate beads when they were administered under fasting conditions with either water or an aqueous solution of citric acid, a potential gut transit delaying substance. The study was performed in healthy male volunteers who swallowed the radio-labelled calcium alginate beads after a 10h overnight fast. Gamma scintigraphy was selected as the method to monitor the movement of the calcium alginate beads. The volunteers consumed no further food or drink until gastric emptying of the calcium alginate beads was complete. The results indicated that prolonged gastric retention was achieved when the dosage form was administered with the citric acid solution when compared to retention in the absence of citric acid. Citric acid, therefore, has the potential to delay the gastric emptying of the calcium alginate beads when administered to fasted volunteers.

Adult↗

The functional evaluation of salivary glands using dynamic MR sialography following citric acid stimulation: a preliminary study.

OBJECTIVE: We introduce a new technique for the functional evaluation of the salivary glands using continuous magnetic resonance (MR) sialography before and after citric acid stimulation. METHODS: In 10 volunteers, the time-dependent changes in the maximum area of the detectable parotid gland ducts on MR sialographic images taken every 30 seconds before and after citric acid stimulation were analyzed. The time period to the occurrence of the maximum duct area poststimulation was noted, and then the time for the area to return to its 50% value pre-citric acid stimulation was also observed. This new technique was clinically applied in 1 patient with an excessive supply impression of saliva and in 1 patient with a short supply impression with saliva. RESULTS: In all volunteers after citric acid stimulation, the maximum area of the detectable salivary gland ducts first increased and then decreased. A strong relationship was found between the maximum area of the detectable salivary gland ducts before citric acid stimulation and total saliva volume (Pearson r = 0.672, P = .031). Compared with all the volunteers, the ratio of change in the detectable ducts was the highest in the patient with an excessive supply impression of saliva, but lowest in the patient with a short supply impression with saliva. CONCLUSIONS: This initial study suggests that dynamic MR sialography allows for functional and morphological evaluation of the salivary glands. This technique appears to have many possible applications and further investigation in this field is necessary.

Adult↗

13C-NMR spectroscopic evaluation of the citric acid cycle flux in conditions of high aspartate transaminase activity in glucose-perfused rat hearts.

A new mathematical model, based on the observation of 13C-NMR spectra of two principal metabolites (glutamate and aspartate), was constructed to determine the citric acid cycle flux in the case of high aspartate transaminase activity leading to the formation of large amounts of labeled aspartate and glutamate. In this model, the labeling of glutamate and aspartate carbons by chemical and isotopic exchange with the citric acid cycle are considered to be interdependent. With [U-13C]Glc or [1,2-(13)C]acetate as a substrate, all glutamate and aspartate carbons can be labeled. The isotopic transformations of 32 glutamate isotopomers into 16 aspartate isotopomers or vice versa were studied using matrix operations; the results were compiled in two matrices. We showed how the flux constants of the citric acid cycle and the 13C-enrichment of acetyl-CoA can be deduced from 13C-NMR spectra of glutamate and/or aspartate. The citric acid cycle flux in beating Wistar rat hearts, aerobically perfused with [U-13C]glucose in the absence of insulin, was investigated by 13C-NMR spectroscopy. Surprisingly, aspartate instead of glutamate was found to be the most abundantly-labeled metabolite, indicating that aspartate transaminase (which catalyses the reversible reaction: (glutamate + oxaloacetate <--> 2-oxoglutarate + aspartate) is highly active in the absence of insulin. The amount of aspartate was about two times larger than glutamate. The quantities of glutamate (G0) or aspartate (A0) were approximately the same for all hearts and remained constant during perfusion: G0 = (0.74 +/- 0.03) micromol/g; A0 = (1.49 +/- 0.05) micromol/g. The flux constants, i.e., the fraction of glutamate and aspartate in exchange with the citric acid cycle, were about 1.45 min(-1) and 0.72 min(-1), respectively; the flux of this cycle is about (1.07 +/- 0.02) micromol min(-1) g(-1). Excellent agreement between the computed and experimental data was obtained, showing that: i) in the absence of insulin, only 41% of acetyl-CoA is formed from glucose while the rest is derived from endogenous substrates; and ii) the exchange between aspartate and oxaloacetate or between glutamate and 2-oxoglutarate is fast in comparison with the biological transformation of intermediate compounds by the citric acid cycle.

Acetyl Coenzyme A↗

Cytotoxic effects of 10% citric acid and EDTA-T used as root canal irrigants: an in vitro analysis.

EDTA-T and 10% citric acid used as root canal irrigants lead to more visible dentinal tubules than 5% sodium hypochlorite associated with 3% hydrogen peroxide. However, these cleansing agents must be compatible with apical periodontal tissue. We analyzed the cytotoxicity of 10% citric acid and EDTA-T in cultured fibroblasts using Trypan blue. The solutions were diluted to 1%, 0.1%, and 0.01% and applied to NIH 3T3 cell cultures. Cells grown on fresh DMEM served as a control. After 0, 6, 12, and 24 h (short-term assay, viability) and 1, 3, 5, and 7 days (long-term assay, survival), the cells were counted using a hemocytometer. In short-term tests, cell viability ranged from 85% to 99% for all experimental groups with no statistical differences when compared with control cultures, except for the group treated with 1% EDTA-T, which caused a progressive decrease in cell viability. In long-term tests, all cultures increased in number from day 1 to the end of the experimental period, showing no inhibition of cell proliferation, except for the cultures treated with 1% EDTA-T, which totally prevented cell growth. All dilutions of 10% citric acid were more biocompatible than EDTA-T. Cultures treated with citric acid had a higher percentage of viable cells in the short-term assays, and the cells retained their self-renewal capacity.

3T3 Cells↗

Comparison of citric acid production by solid-state fermentation in flask, column, tray, and drum bioreactors.

Studies were conducted to evaluate citric acid production by solid-state fermentation (SSF) using cassava bagasse as substrate employing a fungal culture of Aspergillus niger LPB 21 at laboratory and semipilot scale. Optimization of the process parameters temperature, pH, initial humidity, aeration, and nutritive composition was conducted in flasks and column fermentors. The results showed that thermal treatment of cassava bagasse enhanced fungal fermentation efficacy, resulting in 220 g of citric acid/kg of dry cassava bagasse with only treated cassava bagasse as substrate. The results obtained from the factorial experimental design in a column bioreactor showed that an aeration rate of 60 mL/min (3 mL/[g.min]) and 60% initial humidity were optimum, resulting in 265.7 g/kg of dry cassava bagasse citric acid production. This was almost 1.6 times higher than the quantities produced under unoptimized conditions (167.4 g of citric acid/kg of dry cassava bagasse). The defined parameters were transferred to semipilot scale, which showed high promise for large-scale citric acid production by SSF with cassava bagasse. Respirometry assays were carried out in order to follow indirectly the biomass evolution of the process. Citric acid production reached 220, 309, 263, and 269 g/kg of dry cassava bagasse in Erlenmeyer flasks, column fermentors, a tray bioreactor, and a horizontal drum bioreactor, respectively.

Aspergillus niger↗

Phytase, citric acid, and 1alpha-hydroxycholecalciferol improve phytate phosphorus utilization in chicks fed a corn-soybean meal diet.

Previous research from our laboratory has shown that phytase, citric acid, and 1alpha-hydroxycholecalciferol [1alpha-(OH) D3] individually improve phytate P use in young chicks fed a P-deficient corn-soybean meal (C-SBM) diet. The current study was conducted to evaluate combinations of these additives on phytate P utilization. In 3 chick experiments, male crossbred chicks (New Hampshire x Columbian) were fed experimental diets from 8 to 21 d of age. The C-SBM basal diet used in all assays contained no supplemental P and was calculated to provide 23% CP, 0.13% nonphytate P (0.39% total P), 0.62% Ca, 25 mg of cholecalciferol/kg, and 3,260 kcal of TME/kg. In all 3 experiments, factorial arrangements (2 x 2 or 2 x 2 x 2) were used to evaluate 2 levels of phytase (0 and 300 units/kg), citric acid (0 and 3 or 4%), and 1alpha(OH) D3 (0 and 5, 10, or 15 microg/kg). Phytase, citric acid, and 1alpha-(OH) D3 each increased weight gain and tibia ash in all 3 experiments. There were some significant interactions among the compounds, but these were not consistent across experiments. Using standard curve methodology for tibia ash data, it was estimated that 0.03, 0.02, and 0.04% P were released by 3% citric acid, 300 units of phytase/kg, and 5 microg 1alpha-(OH) D3/kg, respectively, and that the combination of all 3 compounds resulted in the release of 0.13% P. Our results indicate that all 3 compounds increased phytate P use, and that their effects were generally additive, with some possible synergism between citric acid and 1alpha-(OH) D3.

6-Phytase↗

Mathematical modeling of citric acid production by repeated batch culture.

A mathematical model has been created for the process of citric acid biosynthesis by yeast (mutant strain Yarrowia lipolytica) cultivated by the repeated batch (RB) method on ethanol under conditions of nitrogen limitation. The model accounts for cell growth as a function of nitrogen concentration in the culture liquid; nitrogen uptake by growing cells; citric acid production; pH control in the fermentor by means of NaOH addition; and changes in system volume. The model represents a system of five nonlinear differential equations. Experimental measurements of cell concentration, citric acid concentration, and cultivation broth volume were used with the least squares method to determine the values of eight model parameters. The parameter values obtained were consistent with literature data and general concepts of cell growth and citric acid biosynthesis. The model has been used to predict optimum RB culture conditions.

Journal Article↗

Resin bonding to Er: YAG laser-irradiated dentin: combined effects of pre-treatments with citric acid and glutaraldehyde.

The purpose of this study was to evaluate the combined effects of citric acid and glutaraldehyde (GA) on the resin bonding to Er: YAG laser-irradiated dentin. Bovine dentin was prepared with 180- to 600-grit SiC paper and then uniformly irradiated with an Er: YAG laser (laser-irradiated group) or immersed in water at 60 degrees C for 15 min (heated group). The samples were then acid-conditioned with 10% citric acid (10-0) or 10% citric acid/3% ferric chloride (10-3) for 15 s and treated with GA for 10 min before bonding to an acrylic rod with 4-META/MMA-TBB resin. These samples were trimmed to prepare miniaturized dumbbell-shaped specimens. After storage in water at 37C for 1 d, the tensile bond strength was measured, and the fractured surface was evaluated using a scanning electron microscope (SEM). In the laser-irradiated and heated groups, the 10-3+GA-treated specimen had higher bond strength than that of 10-0+GA. On the other hand, the tensile bond strength of 10-3 +GA in the non-irradiated group was lower that that of 10-0+GA. In conclusion, the combination of 10-3 and GA for bonding with 4-META/MMA-TBB resin was the most effective for Er: YAG laser-irradiated dentin and heated dentin, but it was not effective for the non-irradiated dentin.

Acid Etching, Dental↗

Citric Acid Fermentation by Aspergillus niger on Low Sugar Concentrations and Cotton Waste.

The possible use of cotton waste as a carbohydrate source of citric acid production by Aspergillus niger was examined. No citric acid was produced when A. niger was grown on cotton waste as a sole carbon source. In two-stage fermentations, however, mycelium obtained from surface cultures in cotton waste medium yielded more citric acid when transferred to sucrose-containing media than when directly inoculated to sucrose-containing media. It is concluded that cotton waste can be used for saving sucrose and for increasing yields of citric acid fermentation by A. niger.

Journal Article↗

Influence of dietary citric acid and calcium on the bioavailability of orally administered chlortetracycline in piglets.

In a study involving 18 piglets divided into three groups (A, B and C), the effects of dietary calcium and citric acid, and feeding technique on chlortetracycline bioavailability were examined. Groups of 6 animals received a basal diet with either 0.7% (group A) or 1.4% calcium (groups B and C). Citric acid was not included in diets of the first experimental period. In the second period, either 1.5% (groups A and B) or 3.0% (group C) citric acid was added to the diet. In both experimental periods, chlortetracycline was administered once intravenously (7.5 mg/kg b.w.) and once orally (30 mg/kg b.w.). After each administration, blood samples were taken at regular intervals in order to determine chlortetracycline serum concentrations by a microbiological method. Following oral chlortetracycline intake, low dietary calcium or citric acid addition to the diet produced increased chlortetracycline serum concentrations. Chlortetracycline bioavailability was 12.6% at 0.7% dietary calcium, and 9.5% at 1.4% dietary calcium. Enteral chlortetracycline absorption was improved 65% with 1.5% or 3.0% dietary citric acid supplementation. An 8-hour delay of feed intake following oral chlortetracycline intake did not significantly influence chlortetracycline bioavailability.

Administration, Oral↗

Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc).

The effects of food components on blood fluidity were studied by in vitro assay using a dedicated microchannel instrument for model capillaries. We found that the fruit-juice concentrate of the Japanese apricot (Prunus mume Sieb. et Zucc), a traditional Japanese food, markedly improved the fluidity of human blood. Using HPLC, we isolated the active compounds and characterized them using UV, MS, IR, and NMR. They included a novel compound, 1-[5-(2-formylfuryl)methyl] dihydrogen 2-hydroxypropane-1,2, 3-tricarboxylate (mumefural), and a related compound, 5-hydroxymethyl-2-furfural (HMF). Mumefural markedly improved blood fluidity in all subjects, while HMF worked differently in different individuals. The flow rate of blood spiked with mumefural or HMF was compared to that of the two predominant organic acids in the fruit. Citric acid, malic acid, and furfuryl alcohol also improved fluidity in all subjects. The activity of P. mume is derived from not only artifacts produced during thermal processing, such as mumefural, but also from endogenous organic acids.

Beverages↗