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Effect of chemical nutrients on aconitase activity during citric acid fermentation by a mutant strain of Aspergillus niger.

An ethyleneimine and UV irradiated mutant strain Aspergillus niger AB 1801 was explored for its aconitase activity, citric acid production and cell growth during citric acid fermentation. 10% sucrose and 0.25% urea served as the best carbon and nitrogen sources, respectively. Fe2+ stimulated aconitase activity and Co2+ (upto 5 micrograms/mL) and Ni2+ (10 micrograms/mL) strongly depressed enzyme activity. Boric acid (0.5 mg/L) had very good stimulatory response towards citric acid production. Aconitase activity has been found to vary inversely with the citric acid production upto a certain limit after which the activity was absent in the mycelial extract. Cell growth varied with the production parameter.

Aconitate Hydratase↗

[Coughing model by microinjection of citric acid into the larynx in guinea pig].

Many studies of cough were performed under the restrained or anesthetized condition, and coughs were evoked by inhalation of capsaicin or citric acid. Inhalation of irritants induced by "diving response" with apnea and coughs, and these responses induced a change of tidal volume. As a result, respiratory responses are dependent on the inhalation volume. Therefore we developed a new coughing model, and coughs were evoked by microinjection of citric acid into the larynx in the unanesthetized unrestrained guinea pig. Microinjection of 7.5% citric acid (2 microliters/30 s, 5 min) induced coughs (27.03 +/- 4.03 coughs/10 min), and citric acid-induced responses were stable independent of the inhalation volume. In the inhalation studies, animals were exposed to citric acid only once because induced-responses were remarkably decreased by repeated administration at an interval of 24 h. However in our coughing model it was possible to repeatedly challenge the animals by microinjection of citric acid at intervals of 24 h. Microinjection of citric acid into the larynx induced coughs in Sprague-Dawley rats, but inhalation of citric acid did not induce cough. These results indicate that this coughing model is highly sensitive and correctly assessed cough responses.

Animals↗

[Effect of citric acid and calcium on the bioavailability of orally-administered oxytetracycline in piglets].

The bioavailability of orally given oxytetracycline in dependence on calcium (0.7% and 1.4% calcium, respectively) and citric acid content in feed was examined in piglets (9.9 +/- 0.9 kg body weight). In the first trial no citric acid was added to the two feeds, in the second trial 1.5% citric acid was added. In both trials each piglet received an i/v-injection of oxytetracycline (dosage 10 mg/kg body weight) and an oral dose of oxytetracycline (40 mg/kg body weight). The blood samples, taken in definite time intervals, were analysed by a microbiological assay. With these results the kinetic parameters and the bioavailability were calculated. The results of the i/v-trials were identical. The kinetic parameters C1 = 14.91 micrograms/ml, lambda 1 = 3.46 h-1, Cz = 7.49 micrograms/ml and lambda z = 0.19 h-1 describe the graph after i/v-application. The calcium content of the feed had no significant influence on the kinetic parameters after p/o-application, but the piglets receiving citric acid showed a significant higher maximum concentration (Cmax). The bioavailability of orally given oxytetracycline was significantly (26%) increased by the citric acid content. In spite of the citric acid the bioavailability only came to 4.9%.

Administration, Oral↗

The effects of citric acid on attachment of laterally positioned flaps.

Citric acid demineralization of root surfaces to enhance new attachment clinically appears to be successful, but histologic evidence of new attachment in humans is lacking. Two recession-like defects were surgically created on the facial aspect of mandibular incisors scheduled for extraction in five patients. Each patient had differing degrees of periodontal disease, but the control and experimental teeth of each were similarly involved. The roots were thoroughly planed and an apical reference notch was inscribed just coronal to crestal bone. One tooth served as the control, while the experimental tooth was treated with citric acid (pH 1) for two minutes. Two full-thickness laterally positioned flaps were done to cover the roots. After healing periods of 1, 2, 4, 12, and 20 weeks, the teeth were removed by block section for histologic evaluation. The type of attachment differed on the teeth with longer healing periods. The control teeth exhibited a long epithelial attachment with no evidence of cementogenesis, whereas the experimental teeth had a connective tissue attachment to newly formed cementum coronal to the reference notch. These observations provide evidence that demineralization of root surfaces may promote increased or accelerated connective tissue attachment of laterally positioned flaps.

Adult↗

High-performance liquid chromatographic measurement of selected blood citric acid cycle intermediates.

We describe a high-performance liquid chromatographic (HPLC) method for analysis of the intermediates of the citric acid cycle. Using two Aminex HPX-87H columns in series at 36 degrees C, the early eluting compounds cis-aconitate, oxaloacetate, alpha-ketoglutarate and citrate-isocitrate can be resolved. Acetonitrile is used for extraction of citric acid cycle intermediates from blood as interfering ultraviolet absorbing peaks are present with perchloric acid or trichloroacetic acid extraction. Acetonitrile extraction is compared with perchloric acid extraction of citric acid cycle intermediates from plasma. Low recovery of some organic acids from blood seems not to be due to enzymatic degradation. Storage of acetonitrile extracts in liquid nitrogen led to a small but significant decrease in pyruvate levels in human blood. However, significant changes in other organic acids were not seen. HPLC methodology allows study of the citric acid cycle in tissue samples as well as blood and promises to facilitate the investigation of human disorders of energy metabolism.

Acetonitriles↗

Extended reuse of polysulfone hemodialysis membranes using citric acid and heat.

The concomitant use of citric acid and prolonged exposure to heat (CAH) is an increasingly common alternative to purely chemical means of reusing dialyzers. However, there are no data on the effects of reprocessing dialyzers with CAH beyond 15 uses. Increasing the number of reuses with CAH cannot be systematically undertaken unless its safety is documented. We hypothesized that discarding polysulfone dialyzers after the 25th rather than the 15th use would result in increased clearance of beta2-microglobulin (beta2MG) without clinically significant changes in small solute clearance or albumin loss. We studied 15 Fresenius F80B polysulfone dialyzers in five chronic hemodialysis patients. Dialyzers were reprocessed using 1.5% citric acid solution heated to 95 degrees C. Representative fractional collection and 10 minute timed collections of dialysate were performed at baseline and during uses 5, 10, 15, 20, and 25 for each dialyzer. Dialysate-side urea, creatinine, and beta2MG clearances were calculated, and total albumin was measured in dialysate. We used a mixed model to adjust for repeated measures (both within a given dialyzer and for the multiple dialyzers per patient). Of the 15 dialyzers studied, 3 (20%) failed before the 25th use. There was no significant change in urea or creatinine clearance with additional reuse (overall p values 0.20 and 0.60, respectively). A sustained increase in beta2MG clearance was observed after the fifth treatment compared with the first use (p < 0.001). Fractional collection showed that dialysate albumin loss increased significantly with additional reuses (p < 0.001) but did not increase significantly above baseline until treatment 25. Reprocessing of polysulfone dialyzers with CAH 25 times significantly increased albumin loss and beta2MG clearance but did not appear to affect urea or creatinine clearance. Increasing the maximum number of uses to 20 may permit cost savings compared with current practice without additional risk.

Adult↗

Evaluation of the effect of EDTA, EDTAC and citric acid on the microhardness of root dentine.

AIM: To evaluate the effect of citric acid, ethylenediaminetetraacetic acid (EDTA) and ethylenediaminetetraacetic acid plus Cetavlon (EDTAC) solutions on the microhardness of human root canal dentine. METHODOLOGY: Sixteen maxillary human canines were sectioned transversely at the cemento-enamel junction and the crowns were discarded. Subsequently, each root was embedded in an epoxy resin cylinder and their middle third sectioned horizontally into 4 mm thick slices. The samples were randomly divided into three groups according to the chelating agent employed, as follows (n = 6): group 1: EDTA 17%, group 2: EDTAC 17% and group 3: citric acid 10%. Dentine microhardness was then measured with a load of 50 g for 15 s. At the beginning of the experiment, reference microhardness values were obtained for samples without any etching (t = 0 min). The same samples were then exposed to 50 microL of the chelator solution for 1, 3 and 5 min. The Student's t-test (P < 0.05) was used to compare results for different times for each chelator and different chelators for each time. RESULTS: Microhardness decreased with increasing time of application of chelating solutions. There were no significant (P > 0.05) differences between initial microhardness for the three groups as well as after 1 min of application of the substances. After 3 min, EDTA produced a significantly greater reduction in microhardness. However, there was no significant difference between EDTA and EDTAC after 5 min. Citric acid caused significantly less reduction in microhardness. CONCLUSIONS: Overall, citric acid was least effective in reducing dentine hardness whilst EDTA had the strongest effect.

Cetrimonium↗

Effect of citric acid on the nucleation of hydroxyapatite in a simulated body fluid.

The role of citric acid in a simulated body fluid (SBF) was examined with the main focus on its induction ability of hydroxyapatite (HAp) nucleation on a bioinert collagen membrane. Collagen membranes were soaked in the SBF with citric acid concentrations in the range of 0-4 mM; then, carbonate-containing HAp crystals grew within a limited range, i.e., 0.3-2 mM. The results were explained in terms of chemical interaction among calcium ion, citric acid and collagen membrane, especially strong chelation ability of citric acid with the calcium ion.

Animals↗

Synthesis and pore formation study of amorphous mesoporous aluminophosphates in the presence of citric acid.

Amorphous mesoporous aluminophosphates (AlPO) with high thermal stability were synthesized from Al(NO3)3 and H3PO4 in the presence of citric acid (CA). The effects of different synthetic conditions, such as pH value, citric acid content, and calcined temperature, on the formation of the mesoporous structure of AlPO materials were investigated. The results of N2 adsorption/desorption and other characterization means demonstrate that the presence of a certain amount of citric acid in the synthetic mixture can result in the formation of uniform mesoporous AlPO materials in a wide pH range from 3.5 to 10. Thermal gravimetry and differential thermal analysis suggest that the existence of a moderate interaction between citric acid and the AlPO network might be a critical factor in obtaining uniform mesostructure. Therefore, as an organic additive, citric acid can be regarded as a template for the mesopore formation of AlPO materials.

Aluminum Compounds↗

Effect of citric acid on soft tissue healing in the rat palate.

The present study was carried out to determine the effects of citric acid (pH 1) on soft tissue healing. Standardised circular wounds were created in the palates of 24 rats using a modified trephine. Test animals had citric acid (pH 1) applied to the wounds for 3 min followed by rinsing with saline. In the control animals saline alone was applied for a similar time which was again followed by rinsing. Animals were sacrificed at 0, 1, 4 and 24 h, the palates dissected out and fixed in either neutral formalin, formol-sucrose for histochemical analysis or had epithelial sheets prepared for ATPase staining of Langerhans cells. At 0, 1 and 4 h less enzyme activity (non-specific esterase and acid phosphatase) was observed in the epithelium in the edge of the citric acid treated wounds compared to the controls. This was not seen at 24 h. No morphological difference was noted between the citric acid treated wounds and the controls at any of the time intervals. The morphology and distribution of ATPase positive Langerhans cells did not differ between the test and control wounds. These results indicate that the application of citric acid (pH 1) has little effect on the short-term healing of soft tissue wounds in the rat palate.

Animals↗

Citric acid production by Aspergillus niger on wet corn distillers grains.

AIMS: To determine which citric acid-producing strain of Aspergillus niger utilized wet corn distillers grains most effectively to produce citric acid. METHODS AND RESULTS: Citric acid and biomass production by the fungal strains were analysed on the untreated grains or autoclaved grains using an enzyme assay and a gravimetric method respectively. Fungal citric acid production on the grains was found to occur on the untreated or autoclaved grains. The highest citric acid level on the grains was produced by A. niger ATCC 9142. The autoclaved grains supported less citric acid production by the majority of strains screened. Biomass production by the fungal strains on the untreated or autoclaved grains was quite similar. The highest citric acid yields for A. niger ATCC 9142, ATCC 10577, ATCC 11414, ATCC 12846 and ATCC 26550 were found on the untreated grains. Treatment of the grains had little effect on citric acid yields based on reducing sugars consumed by A. niger ATCC 9029 and ATCC 201122. CONCLUSIONS: It is feasible for citric acid-producing strains of A. niger to excrete citric acid on wet corn distillers grains whether the grains are treated or untreated. The most effective citric acid-producing strain of A. niger was ATCC 9142. SIGNIFICANCE AND IMPACT OF THE STUDY: The study shows that the ethanol processing co-product wet corn distillers grains could be utilized as a substrate for the commercial production of citric acid by A. niger without treatment of the grains.

Aspergillus niger↗

Effects of aluminum sulphate and citric acid ingestion on lipid peroxidation and on activities of superoxide dismutase and catalase in cerebral hemisphere and liver of developing young chicks.

Effect of oral administration of aluminum sulphate (200 and 400 mg/kg body wt/day) without or with citric acid (62 mg/kg body wt/day) to day-old White Leghorn male chicks (n = 5 per group) for 30 days was studied on the activities of superoxide dismutase (SOD) and catalase, and level of lipid peroxidation in cerebral hemisphere and liver. A 400 mg dose of Al in the presence of citric acid inhibited cytosolic total and CN -sensitive superoxide dismutase activities of the cerebral hemisphere in 7- and 30-day treated chicks, whereas in 15-day treated chicks the enzyme activities were decreased in response to both doses in the presence of citric acid. In case of liver, activities of these enzymes significantly decreased after 7, 15 and 30 days of treatment with 200 and 400 mg Al together with citric acid, whereas 400 mg Al alone inhibited the enzyme activities after 15 and 30 days of treatment. Cerebral catalase activity decreased in response to 400 mg Al when the chicks were also fed with citric acid for 7 and 30 days, but in 15-day treated chicks the enzyme activity was depleted following treatment with 200 and 400 mg Al combined with citric acid. 400 mg Al treatment for 7 days in combination with citric acid inhibited hepatic catalase activity and extension of the treatment period to 15 and 30 days also produced reduction in its activity even in response to the lower Al dose mixed with citric acid. CN -insensitive SOD activity of cerebral hemisphere and liver was unaffected by Al. Al also failed to induce lipid peroxidation in both the tissues throughout the course of exposure. Activities of SOD and catalase of cerebral hemisphere and liver of 30-day old chicks were observed to be inhibited by in vitro incubation with different concentrations of Al. Our in vivo study demonstrates that only CN -sensitive SOD is susceptible to Al. Further, responses of SOD and catalase to Al is tissue specific. The observed inhibition of antioxidant enzyme activities by Al is suggestive of a prooxidant state. Induction of such an oxidative condition of the tissues may be attributed to a direct effect of the metal on enzyme molecules or in their synthesis.

Alum Compounds↗

Citric acid or citrates in urine: which should we focus on in the prevention of calcium oxalate crystals and stones?

In order to distinguish between normocitraturia and hypocitraturia, the 24 h urine excretion value of citric acid is evaluated in relation to the established limit value of 2.5 mmol/day. We propose changing this widely-used excretion value to a "minimum contribution" of citric acid to the total urine's ionic strength, since the inhibitory effect of citric acid on crystallization depends on citrate anions being available to complex calcium ions or to associate with the crystal surface. A total of 71424 h-urine samples, taken from 74 healthy persons and 58 calcium stone formers, were investigated for pH, citric acid concentration ([CA]), and related relative calcium oxalate supersaturation (RS). Based on the Henderson-Hasselbalch-equation, the individual concentrations of the differently charged citrate anion species in each of the urines were calculated from the urinary pH and [CA]. From the anion concentrations determined, the contribution of the urine's citric acid to the total urine's ionic strength, ISCA, was calculated. Referring to the limit value of 2.5 mmol/day and assuming an average urine volume of 1.5 l/day, a hypothetical concentration limit of 1.67 mmol/l can be obtained. Grouping the samples into "stone-formers" and "non-stone-formers" as well as into three different ranges of RS revealed: (1). that the groups' median [CA]-values were below 1.67 mmol/l, and (2). that [CA] was not inversely associated with the risk of stone formation. Within the pH-range of 5 and 7, the ISCA-values which are related to, for example, [CA]=1.67 mmol/l, vary considerably by a factor of nearly three between 2.48 mmol/l and 6.64 mmol/l. The use of a fixed citric acid excretion level for the distinction of normocitraturia from hypocitraturia does not take into account the different citrate species which actually modify the urine's crystallization behaviour. The proposed ISCA approach takes this fact into consideration. From this parameter, a desirable "minimum impact of citric acid" can be derived. In a first approach, a potential ISCA-limit value, which currently distinguishes between urines indicated by a "normo-protective" impact and those indicated by a "hypo-protective" impact with respect to calcium oxalate precipitation, may be set at 2.48 mmol/l.

Adult↗

[The effect of a supplement of citric acid on the bioavailability of zinc from corn germ].

The purpose of this 2 factorial designed study was to investigate the influence of citric acid on the availability of zinc from diets containing 140 g corn germs as a native phytate source (0.5% phytate in diet). Growing male rats with an average initial weight of 42 g were divided into 8 groups of 8 animals each. After a 7 d depletion period (2.4 micrograms Zn/g diet) the animals were fed ad libitum for 21 d a diet on the basis of egg white solid and corn germs. The diets were supplemented with zinc in order to obtain phytate:zinc molar ratios of 31, 20, 14, and 0 (control without corn germs, 11 micrograms Zn/g diet). Each diet was fed with and without a supplementation of 1% citric acid. A phytate:Zn molar ratio of 31:1 resulted in typical symptoms of zinc-deficiency like anorexia, alopecia and a significant depression of growth. These effects were apparently reduced by citric acid. The zinc concentration in serum and organs followed the graded levels of phytate:zinc molar ratios. Primary significant effects of the phytate:Zn molar ratio but also effects of citric acid and interactions between the 2 factors phytate:Zn and citric acid could be detected. Only total liver zinc but not liver zinc based on fresh matter was affected by the phytate:Zn molar ratio. In serum and tissues the activity of alkaline phosphatase showed a significant response to the phytate:zinc molar ratio. Furthermore the supplementation with citric acid increased the femur alkaline phosphatase and slightly reduced it in the liver. The concentrations of metallothionein in liver duodenum, jejunum and ileum were significantly affected by the phytate:Zn molar ratio.

Alkaline Phosphatase↗

Healing following reimplantation of teeth subjected to root planing and citric acid treatment.

The aim of the present study was to examine the effect of citric acid treatment on periodontal healing around teeth which were extracted, root planed and then reimplanted. Maxillary incisors and mandibular incisors, premolars and molars of 5 monkeys were used. The teeth were divided into 3 experimental groups. In 1 group, the teeth were extracted and immediately reimplanted into their own sockets. In a 2nd group, the teeth were extracted, root planed to a level corresponding to 50-75% of the root length and then reimplanted. In the 3rd group, periodontal breakdown extending to 50-75% of the root length was first induced by placing orthodontic elastic ligatures around the teeth. They were then extracted and root planed and transplanted into the sockets of the contralateral, periodontally healthy teeth which had just been extracted. Half the number of the teeth of groups 2 and 3 were treated with citric acid before reimplantation or transplantation. The animals were sacrificed after 6 months of healing. The jaws were removed and histological specimens prepared for microscopic examination. With the exception of a limited coronal regrowth of new cementum in the apical part of the planed portion of a few roots, connective tissue attachment failed to reform on most root surfaces deprived of their periodontal ligament tissue. Healing was most frequently characterized by root resorption and ankylosis. These were the most predominant features of healing both on root surfaces which had been deprived of the ligament tissue by mechanical means or during a course of experimentally-induced periodontal disease, and occurred in citric acid as well as non-citric acid treated roots. The findings imply that the result of healing following tooth reimplantation or transplantation is determined by the type of cells that repopulate the wound area adjacent to the denuded root surface.

Animals↗

Yarrowia lipolytica as a potential producer of citric acid from raw glycerol.

AIMS: To study the biochemical response of Yarrowia lipolytica LGAM S(7)1 during growth on raw glycerol (the main by-product of bio-diesel production units) in order to produce metabolic products of industrial significance. METHODS AND RESULTS: Yarrowia lipolytica was cultivated on raw glycerol or glucose in flasks. Although nitrogen-limited media were employed, growth was not followed by production of reserve lipid. Nitrogen limitation led to citric acid excretion. Growth and citric acid production parameters on glycerol were similar to those obtained on glucose. When high initial glycerol media were used, citric acid up to 35 g l(-1) (yield 0.42-0.44 g acid g(-1) glycerol consumed) was produced. CONCLUSIONS: Raw glycerol was an adequate substrate for Y. lipolytica. Growth was not followed by reserve lipid accumulation, but amounts of citric acid were produced. SIGNIFICANCE AND IMPACT OF THE STUDY: Raw glycerol is an industrial feedstock appearing in increasing quantities as the main by-product of bio-diesel production facilities. The present study describes an alternative way of glycerol valorization, with the production of remarkable amounts of citric acid, in addition to its main valorization way (production of 1,3-propanediol by bacteria).

Biomass↗

Fed-batch production of citric acid by Candida lipolytica grown on n-paraffins.

This study reports on the effects of fermentor agitation and fed-batch mode of operation on citric acid production from Candida lipolytica using n-paraffin as the carbon source. An optimum range of agitation speeds in the 800-1000 rpm range corresponding to Reynolds numbers of 50000-63000 (based on initial batch conditions) seemed to give the best balance between substrate utilization for biomass growth and citric acid production. Application of multiple fed-batch feedings can be used to extend the batch fermentation and increase final citric acid concentrations and product yield. The three-cycle fed-batch system increased overall citric acid yields to 0.8-1.0 g citricacid/g n-paraffin, approximately a 100% improvement in product yield from those observed in the single cycle fed-batch system and a 200% improvement over normal batch operation. The three-cycle fed-batch mode of operation also increased the final citric acid concentration to 42 g/l from about 12 and 6g/l for single fed-batch cycle and normal batch modes of operation, respectively. Increased citric acid concentrations in three-cycle fed-batch mode was achieved at longer fermentation times.

Biomass↗

[Cough and bronchial obstruction induced by citric acid in smokers, occasional smokers and non-smokers].

We have studied cough and bronchial constriction induced by inhaling citric acid in 15 smokers with baseline airflow obstruction, in 13 occasional smokers and 13 non smokers. The threshold for cough was significantly higher in occasional smokers in relation to smokers and non smokers. Citric acid produced the same degree of bronchial constriction at the same time in smokers and occasional smokers: the maximum fall in forced expired volume (FEV1-VEMS) was recorded five seconds after inhalation of the citric acid (dose threshold) and there was no significant difference between the two groups. In the non smokers, the maximum fall in VEMS was recorded twenty seconds after inhalation of the citric acid and was significantly less in relation to that of the smokers and occasional smokers. In the smokers the degree of smoking could influence the fall of VEMS (% predicted). Cough and bronchial constriction after the inhalation of citric acid are probably related to different physiological mechanisms and are linked to the history of smoking.

Adult↗