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Effect of varying doses of potassium-magnesium citrate on thiazide-induced hypokalemia and magnesium loss.

The purpose of this study was to compare the efficacy of three dosages of potassium-magnesium citrate in overcoming thiazide-induced hypokalemia and magnesium loss and increasing urinary pH and citrate. Sixty-one normal subjects first took hydrochlorothiazide at 50 mg/d. After 3 weeks of thiazide treatment or earlier if hypokalemia developed, the subjects were randomized to take one of three dosages of potassium-magnesium citrate (K ( 4 ) MgCit ( 2 ) ) for 3 weeks while continuing on the thiazide: 4 tablets per day (24 mEq potassium, 12 mEq magnesium, and 36 mEq citrate per day), 7 tablets per day (49 mEq potassium, 24.5 mEq magnesium, and 73.5 mEq citrate per day), or 10 tablets per day (70 mEq potassium, 35 mEq magnesium, and 105 mEq citrate per day). Outcome measures were changes in serum potassium and magnesium and urinary potassium, magnesium, pH, and citrate. All three dosages of potassium-magnesium citrate significantly increased serum potassium concentration, with >80% of subjects regaining normal values despite continued thiazide therapy. The two higher dosages, but not the lowest dosage, caused a small but significant increase in serum magnesium concentration, while substantially increasing urinary magnesium. All three dosages significantly increased urinary pH and citrate in a dose-dependent manner. The lowest dosage produced increases sufficient to prevent stone recurrence. Side effects of thiazide therapy were ameliorated by the highest dosage but not by the two lower dosages. Potassium-magnesium citrate at a dosage of 4 tablets per day is adequate to correct thiazide-induced hypokalemia and to increase urinary pH and citrate sufficiently for stone prevention. Higher dosages are probably required for the prevention of magnesium loss and adverse symptoms of thiazide therapy.

Adult↗

Effects of pH, calcium, and succinate on sodium citrate cotransport in renal microvilli.

Sodium-dependent citrate uptake (v) by rabbit renal brush border vesicles was studied as a function of pH and Ca2+, citrate, and succinate concentrations to evaluate the hypothesis that factors which alter renal citrate reabsorption also alter v. At pH 8 a 10-fold increment in citrate concentration (up to 3.3 mM) was required to achieve the same v observed at pH 7. This provides proof that the protonated citrates are the effective substrates for v. At higher citrate concentrations (5-10 mM) and pH 8, substrate inhibition of v occurred. Studies of the substrate concentration dependency of v at various pH levels and calcium concentrations indicated that v was competitively inhibited by citrate3-. Changes in calcium concentrations altered v by altering the protonated citrate and citrate3- concentrations; v showed countertransport with succinate. Sodium-dependent succinate uptake (vs) was not pH dependent but was more inhibited by 1 mM citrate at pH 7 than at pH 8, indicating that the protonated citrates are inhibitors of vs. However, citrate3- was also found to inhibit vs. Thus, changes in v at the brush border membrane due to the effects of pH or calcium concentration on the protonated citrate concentration and to competition between polycarboxylates may be relevant to changes in renal citrate reabsorption in vivo.

Animals↗

Dietary citrate treatment of polycystic kidney disease in rats.

Progression of autosomal-dominant polycystic kidney disease (ADPKD) in the heterozygous male Han:SPRD rat is dramatically slowed by ingestion of potassium or sodium citrate. This study examined the efficacy of delayed therapy with sodium citrate, the effect of sodium citrate therapy on kidney cortex levels of transforming growth factor-beta (TGF-beta), and the response to calcium citrate ingestion. Rats were provided with citrate salts in their food, and renal clearance, blood pressure, blood chemistry, and survival determinations were made. Sodium citrate therapy was most effective when started at age 1 month, and delay of therapy until age 3 months produced no benefit. Kidney cortex TGF-beta levels were elevated in 3- and 8-month-old rats with ADPKD, but not in 6-week-old rats. Sodium citrate treatment, started at age 1 month, lowered TGF-beta levels to normal in 3-month-old rats, but this is probably not the primary mechanism of citrate's beneficial effect. Calcium citrate had only a modest effect in preserving glomerular filtration rate. Effective treatment of ADPKD in this rat model requires early administration of a readily absorbed alkalinizing citrate salt. Existing data on ADPKD patients on vegetarian diets or with kidney stones should be studied in light of these findings.

Animals↗

Does citrate prevent nephrocalcinosis in preterm neonates?

Nephrocalcinosis (NC) occurs frequently in preterm neonates. A high U-calcium/citrate is one of the contributing factors to the development of NC. In stone-forming children and adults citrate supplementation is a successful preventive therapy. In this randomized controlled trial the effect of citrate therapy was studied on the development of NC in preterm neonates with a gestational age <32 weeks. Thirty-eight preterm neonates (mean gestational age 29.8 weeks (SD 1.6), mean birth weight 1,300 g (SD 351) were treated with sodium citrate (0.52 mmol/kg/day in four doses) from day 8 of life until at term and 36 preterm neonates (mean gestational age 29.6 weeks (SD 1.6), mean birth weight 1,282 g (SD 256) were not treated. U-calcium, U-creatinine, U-citrate and U-pH were measured at day 7, 14, 21, 28 of life and at term. Renal ultrasonography (US) was performed at term. U-citrate/creatinine and U-pH were significantly higher and U-calcium/citrate was significantly lower in the citrate group at day 14, 21 and 28 compared with the control group (P<0.05). Complications of citrate administration were not encountered, however the incidence of NC was not significantly different in the treated (34%) compared with the control group (44%), P=0.37. Preterm neonates treated with citrate in the first months of life have higher U-citrate/creatinine and lower U-calcium/citrate compared with controls. Sodium citrate therapy in a dosage of 0.52 mmol/kg/day is safe but does not prevent NC. Whether a higher dose or potassium citrate decreases the incidence of NC should be evaluated in further studies.

Citric Acid↗

Urinary citrate in preterm and term babies.

OBJECTIVES: (1) To determine a normal range for urinary citrate for term babies. (2) To compare urinary citrate measured in ex preterm babies at term with this normal range. (3) To evaluate whether urinary citrate was related to presence of nephrocalcinosis (NC) and chronic Lung Disease (CLD) in these ex preterm babies. STUDY DESIGN: Urinary citrate was measured in 38 healthy term babies (mean birth weight 3.52 kg, mean gestation 41 weeks) at a mean postnatal age of 3 days (1-5 days) and in 53 ex preterm babies (<32 weeks gestation at birth) at term. These preterm babies were part of a larger study on NC in which two renal ultrasound scans were performed at 1 month and term. RESULTS: The normal range for urinary citrate in term babies was 0.025-2.97 (mean 1.03) mmol/l and citrate/creatinine ratio 0.0011-0.852 (mean 0.27). In the ex-preterm urinary citrate was not significantly different (mean 1.1 vs. 1.03, p=0.7232) but urine citrate /creatinine ratio was significantly higher (mean 1.27 vs. 0.27, p=0.0005). There was no significant difference in urinary citrate or ratios of citrate/creatinine and calcium/citrate in the 11 (20.7%) with NC or in the 17 (32%) babies with CLD. There was no significant relationship found between duration of TPN and urinary citrate measured at term. CONCLUSION: We have determined a normal range for urinary citrate in healthy term babies in the first week of life. The range was very wide. Ex preterm babies had similar values at term and there was no association between urinary citrate and NC or CLD.

Calcium↗

Citrate ions enhance behavioral and cellular responses to taste stimuli.

Behavioral and electrophysiological experiments have been performed on male Sprague-Dawley rats to evaluate the effectiveness of citrate ions as taste enhancers. In two- and four-day two bottle preference tests (vs. water), citrate (1-25 mM) significantly enhanced preference for sweet compounds and the amino acid glycine over control (non-citrate containing) solutions. Under conditions in which animals were "forced" to choose between test solutions with or without citrate, saccharin, sucrose, glycine and NaCl solutions containing citrate were significantly preferred over controls. In addition to its effects on sweet, glycine and NaCl taste, short term preference tests following water deprivation revealed that citrate modulated acid taste preference, as well. Bitter taste preference remained insensitive to citrate in all behavioral assays. As a preliminary attempt to elucidate the target of citrate's actions, electrophysiological responses of isolated fungiform taste receptor cells (TRCs) to saccharin and glycine were recorded using patch clamp techniques. Saccharin (20 mM) and glycine (50 mM) elicited action potentials from TRCs in current clamp mode. Addition of citrate caused a significant increase in number of action potentials generated per 30 s stimulation. Citrate caused a slight but significant depolarization from resting potentials in most TRCs, independent of tastant effects. Taken together, these results suggest that citrate acts at the receptor cell level to enhance only those responses to tastants which depolarize TRCs (i.e. sweet, salt, glycine, acid) while leaving unaffected those which do not depolarize TRCs (bitter).

Animals↗

Citrate plasma levels in patients under regional anticoagulation in continuous venovenous hemofiltration.

BACKGROUND: Different methods of regional anticoagulation using citrate in continuous hemofiltration have been described. To date, only such surrogate parameters as pH, anion gap, total calcium concentration, or total calcium-ionized calcium ratio have been proposed to reflect increased plasma citrate levels and thus risk for side effects. However, none of these parameters has been correlated with plasma citrate levels in critically ill patients. METHODS: Sixteen patients were treated with continuous venovenous hemofiltration (CVVH) and citrate anticoagulation for a mean of 13 +/- 9 days. Citrate levels were measured every other day, and correlations were calculated with the mentioned parameters. RESULTS: Steady-state citrate levels on treatment day 3 were 16.39 +/- 15.77 mg/dL (range, 2.63 to 73.49 mg/dL [853 +/- 821 micromol/L; range, 137 to 3825 micromol/L]). The highest correlation was found between citrate plasma level and total calcium-ionized calcium ratio (R = 0.85; P < 0.001). pH (R = -0.15) and anion gap (R = 0.36) were not helpful in estimating citrate plasma levels in patients treated with citrate-CVVH. CONCLUSION: Calculating total calcium-ionized calcium ratio is a simple tool that correlates best with citrate plasma levels. We recommend close monitoring of this parameter in all patients administered high doses of citrate as part of regional anticoagulation protocols.

Acute Kidney Injury↗

An excess of topical calcium and magnesium reverses the therapeutic effect of citrate on the development of corneal ulcers after alkali injury.

Our purpose was to determine whether chelation of Ca2+ and Mg2+ is the mechanism by which sodium citrate inhibits corneal ulceration in the alkali-injured rabbit eye. The right eyes of 60 albino rabbits (2-2.5 kg) were alkali-injured by filling a 12-mm-diameter plastic well placed on the corneal surface with 0.4 ml of 1 N NaOH. After 35 s the alkali was aspirated, and the well was rinsed with physiological saline. Animals were randomly distributed to three treatment groups of equal size. Two drops of the following topical medications were administered on the hour (14 times per day) for 35 days: physiological saline, 10% citrate in saline, and 346 mM Ca2+, 346 mM Mg2+, and 10% citrate in saline. During the experiment, significantly fewer ulcerations occurred in the citrate-treated eyes (five of 20, 25%) than in the saline-treated eyes (13 of 20, 65%) or in the calcium-magnesium-citrate-treated eyes (15 of 20, 75%). When ulcerations did develop in the citrate group, they occurred significantly later and were less severe than those in the saline and calcium-magnesium-citrate groups. There was a significant increase in the number of eyes with signs of band keratopathy and translucent areas in the calcium-magnesium-citrate group when compared with the other two groups. As in previous studies, sodium citrate significantly inhibited the development of corneal ulcers after alkali injury. The annullment of the favorable effect of citrate on ulceration in the alkali-injured eye by the addition of calcium and magnesium shows that the mechanism of action of citrate is the chelation of these divalent cations.

Administration, Topical↗

Citrate Metabolism by Pediococcus halophilus.

Several strains of non-citrate-metabolizing Pediococcus halophilus have previously been isolated from soy sauce mash or moromi. The factors controlling the metabolism of citrate in soy pediococci were studied. All the soy pediococcal strains tested which failed to decompose citrate did not possess citrate lyase [citrate (pro-3S)-lyase; EC 4.1.3.6] activity. In P. halophilus, citrate lyase was an inducible enzyme, and the optimum pH for activity was 7.0. The metabolism of citrate in P. halophilus was different from that observed in lactic streptococci. The main products from citrate were acetate and formate, and this bacterium produced no acetoin or diacetyl. Formate production from citrate was greatly reduced in the presence of glucose. P. halophilus 7117 (Cit) was proved to contain citrate lyase, pyruvate formate-lyase (EC 2.3.1.54) phosphotransacetylase (phosphate acetyltransferase; EC 2.3.1.8), and acetate kinase (EC 2.7.2.1), i.e., all the enzymes necessary to convert citrate to acetate and formate.

Journal Article↗

Impairment of citrate metabolism in acute hepatic failure.

AIMS: To compare the utilization of citrate employed as anticoagulant in patients with acute hepatic failure and subjects with normal liver function. PATIENTS AND METHODS: Three patients in acute hepatic failure and normal renal function were studied during therapeutic plasma exchange with citrate containing fresh frozen plasma. Six patients receiving immunapheresis or LDL-apheresis anticoagulated with citrate served as controls. Determinations of serum citrate concentrations, of ionized calcium and blood pH were performed before, during, and after the extracorporeal treatment. Total body clearance and elimination half life were calculated in a two compartment model. RESULTS: Preinfusion citrate levels were higher in the patients with acute hepatic failure than in the controls (n.s.). The citrate level rose to 1.73 +/- 0.2 mmol/l in the liver patients versus 0.99 +/- 0.1 mmol/l in the healthy subjects (p < 0.03). Total body clearance was markedly reduced in patients with acute hepatic failure (3.31 +/- 0.03 ml/kg/min) as compared with the controls (6.34 +/- 0.16 ml/kg/min) (p < 0.02), the elimination half life (t/2 k1e) was prolonged (49.7 +/- 5.4 vs. 32.9 +/- 1.02 min, p < 0.05). In the controls blood pH rose from 7.4 +/- 0.01 to 7.45 +/- 0.01 (p < 0.05) after citrate infusion, whereas in the liver patients no rise in pH was observed, again reflecting the impairment of citrate metabolism. Ionized calcium was lower in the patients with acute hepatic failure at the beginning (1.01 +/- 0.05 vs. 1.21 +/- 0.04 mmol/l, p < 0.05) and the end (0.68 +/- 0.02 vs. 0.93 +/- 0.04 mmol/l, p < 0.05) of the citrate infusion. CONCLUSIONS: Citrate metabolism is severely impaired and the plasmatic calcium stores are reduced in acute hepatic failure and, thus, the risk of adverse effects is high. Therapeutic infusions of citrate should be restricted in patients with acute hepatic failure and, if necessary, therapy should be closely monitored by repeated measurements of ionized calcium to avoid the development of potentially hazardous hypocalcemia.

Adolescent↗

Suppression of agglomeration in fluidized bed coating. IV. Effects of sodium citrate concentration on the suppression of particle agglomeration and the physical properties of HPMC film.

We previously reported that sodium citrate (Na citrate), which is a high order salt in the Hofmeister's series, greatly suppressed particle agglomeration in fluidized bed coating (Pharm. Res., 16 (1999), 1616-1620). In this paper, we studied the effects of Na citrate concentration on the particle agglomeration in fluidized bed coating and on the structure of coated film on the particles. Spherical granules made of crystalline cellulose (Celphere) containing phenacetin were coated in a fluidized bed with the aqueous coating solution of hydroxypropylmethyl cellulose (HPMC) containing Na citrate at various concentrations. The particle diameter and drug release profile of coated particles, and the physical properties, i.e. tensile strength, elongation percentage at break, porosity and pore size distribution, of the HPMC cast film were investigated. The particle agglomeration was suppressed with the increasing Na citrate concentration. It is considered that the increase in the suppression effect was caused by the salting-out effect of the increased Na citrate. In the HPMC cast film system, the tensile strength and elongation percentage decreased and the porosity and cumulative pore volume increased with an increase in Na citrate concentration. It is considered that the increase in the porosity by adding Na citrate resulted from a phase separation due to the salting-out during the film forming process. The drug release rate from coated particles also increased with the increasing Na citrate concentration. It can be concluded that the increase in the release rate was due to the increase in porosity of the HPMC coated film caused by the increased Na citrate concentration.

Algorithms↗

Citrate therapy for polycystic kidney disease in rats.

BACKGROUND: Few treatments are available to slow the progression to renal failure in autosomal dominant polycystic kidney disease (PKD). In an animal model of PKD, the male heterozygous Han:SPRD rat, intake of a solution of potassium citrate plus citric acid (KCitr) from age one to three months prevented a decline in glomerular filtration rate (GFR). The present study tested whether this beneficial effect is sustained and explored handling of citrate and ammonia in normal and cystic kidneys. METHODS: Rats were provided with tap water or citrate solutions to drink, and clearance and survival studies were performed. RESULTS: The GFRs of rats with PKD that consumed KCitr from one month of age were normal at six months of age, while those of their counterparts on water were about one third of normal. Long-term KCitr treatment extended the average life span of rats with PKD from 10 to 17 months. Compared with normal rats, water-drinking rats with PKD had higher plasma [citrate], renal cortical [citrate], and fractional excretion of citrate, and lower rates of renal citrate consumption, ammonia synthesis, and ammonia excretion. Cortical PNH3 was not elevated in cystic kidneys. Intake of Na3 citrate/citric acid solution or K3 citrate solution, but not ammonium citrate/citric acid solution, prevented a decline in GFR in three-month-old rats with PKD. CONCLUSIONS: Rats with PKD show abnormal renal handling of citrate and ammonia. Citrate salts that have an alkalinizing effect preserve GFR and extend survival.

Ammonia↗

A continuous, spectrophotometric activity assay for nitrogenase using the reductant titanium(III) citrate.

A continuous, spectrophotometric assay for determining electron transfer rates through nitrogenase during substrate reduction reactions was developed. The assay takes advantage of the facts that Ti(III) citrate can serve as a reductant for nitrogenase-catalyzed reduction reactions and that oxidation of Ti(III) citrate to Ti(IV) citrate results in a dramatic change in its absorption spectrum. Ti(III) citrate supported nitrogenase-catalyzed substrate (e.g., H+ or acetylene) reduction reactions at about the same rate as that supported by the reductant dithionite (S2O4(2-)). In addition, Ti(III) citrate had an absorption maximum centered at 325 nm, while oxidized Ti(IV) citrate had a much lower absorption in this wavelength region. An absorption coefficient for Ti(III) citrate of 0.73 mM-1.cm-1 at 340 nm was determined by titration with redox dyes with known absorption coefficients. Using this experimentally determined absorption coefficient, we developed an assay that provides a convenient way to determine electron transfer rates through nitrogenase in real time by spectrophotometrically following the oxidation of Ti(III) citrate to Ti(IV) citrate. Average electron transfer rates of 3749 +/- 218 nmol of electrons transferred.min-1.mg iron protein-1 for H+ reduction were determined using this assay which are directly comparable to the rates calculated from fixed time point, gas chromatographic assays of H2 formation. The utility of the Ti(III) citrate assay for nitrogenase is discussed and demonstrated using the nitrogenase inhibitors MgADP, CN-, and NO.

Azotobacter vinelandii↗

Induction of citrate synthase by aldosterone in the rat kidney.

The possible induction of renal citrate synthase (E.C. 4.1.3.7) by aldosterone was evaluated in the adrenalectomized rat. Three hours after administration of aldosterone (0.8 microgram/100 g body wt), renal cortical and medullary citrate synthase activity was significantly increased as reported previously by Kinne and Kirsten (Kinne, R., Kirsten, R. 1968. Pfleugers Arch. 300:244). In contrast, no change in this activity was detected in the renal papilla or the liver, under the same conditions. Kinetic analysis revealed that injection of aldosterone had no effect on the KmS for acetyl-CoA and oxalacetate but augmented Vmax of renal medullary citrate synthase activity by 40%. The aldosterone-dependent increase in medullary citrate synthase activity was proportionate to the associated increase in the quantity of antiserum (specific for citrate synthase) required for half-maximal immuno-precipitation. The possibility that aldosterone induced the synthesis of citrate synthase was evaluated in two sets of experiments. In the first set, adrenalectomized rats were injected intraperitoneally with either aldosterone (0.8 microgram/100 g body wt) or the diluent, and simultaneously with 3H or 35S methionine (500 muCi/rat). The isotopes were reversed in about half of the experiments. Three hours after the injection, renal citrate synthase was isolated by ATP-sepharose column chromatography and immuno-precipitation with the specific antiserum. Aldosterone augmented methionine incorporation into renal citrate synthase by 55% but had no effect on incorporation into the hepatic enzyme. In the second set, adrenalectomized rats were injected with either aldosterone (0.8 microcram/100 g body wt) or the diluent, the kidneys were removed 1 hr later and medullary slices were incubated in either 3H- or 35S-methionine at 20 degrees for 2 hr. Mitochondrial citrate synthase was isolated either by ATP-sepharose column chromatography and immuno-precipitation, or by polyacrylamide gel electrophoresis. Aldosterone increased methionine incorporation into the immuno-precipitates by 30% and into the enzyme peak resolved by polyacrylamide gel electrophoresis by 43%. The latter increase was eliminated by prior administration of either actinomycin D (70--80 microgram/100 g body wt) or spirolactone (SC-26304) (80 microgram/100 g body wt). An equimolar dose of dexamethasone (0.8 microgram/100 g body wt) had no effect on the isotope ratio associated with citrate synthase activity in the polyacrylamide gels.

Acetyl Coenzyme A↗

An investigation into the mechanism of citrate-Fe2+-dependent lipid peroxidation.

Chelation by citrate was found to promote the autoxidation of Fe2+, measured as the disappearance of 1,10-phenanthroline-chelatable Fe2+. The autoxidation of citrate-Fe2+ could in turn promote the peroxidation of microsomal phospholipid liposomes, as judged by malondialdehyde formation. At low citrate-Fe2+ ratios the autoxidation of Fe2+ was slow and the formation of malondialdehyde was preceded by a lag phase. The lag phase was eliminated by increasing the citrate-Fe2+ ratio, which also increased the rate of Fe2+ autoxidation. The Fe2+ autoxidation product required for the initiation of lipid peroxidation was characterized as being Fe3+. As direct evidence of this, linear initial rates of lipid peroxidation were obtained via the combination of citrate-Fe2+ and citrate-Fe3+, optimum activity occurring at a Fe3+-Fe2+ ratio of 1:1. Evidence is also presented to suggest that the superoxide and the hydrogen peroxide that are formed during the autoxidation of citrate-Fe2+ can either stimulate or inhibit lipid peroxidation by affecting the yield of citrate-Fe3+ from citrate-Fe2+. No evidence was obtained for the participation of the hydroxyl radical in the initiation of lipid peroxidation by citrate-Fe2+.

Animals↗

Urinary citrate in preterm and term babies.

OBJECTIVES: (1) To determine a normal range for urinary citrate for term babies. (2) To compare urinary citrate measured in ex preterm babies at term with this normal range. (3) To evaluate whether urinary citrate was related to presence of nephrocalcinosis (NC) and chronic Lung Disease (CLD) in these ex preterm babies. STUDY DESIGN: Urinary citrate was measured in 38 healthy term babies (mean birth weight 3.52 kg, mean gestation 41 weeks) at a mean postnatal age of 3 days (1-5 days) and in 53 ex preterm babies (<32 weeks gestation at birth) at term. These preterm babies were part of a larger study on NC in which two renal ultrasound scans were performed at 1 month and term. RESULTS: The normal range for urinary citrate in term babies was 0.025-2.97 (mean 1.03) mmol/l and citrate/creatinine ratio 0.0011-0.852 (mean 0.27). In the ex-preterm urinary citrate was not significantly different (mean 1.1 vs. 1.03, p=0.7232) but urine citrate/creatinine ratio was significantly higher (mean 1.27 vs. 0.27, p=0.0005). There was no significant difference in urinary citrate or ratios of citrate/creatinine and calcium/citrate in the 11 (20.7%) with NC or in the 17 (32%) babies with CLD. There was no significant relationship found between duration of TPN and urinary citrate measured at term. CONCLUSION: We have determined a normal range for urinary citrate in healthy term babies in the first week of life. The range was very wide. Ex preterm babies had similar values at term and there was no association between urinary citrate and NC or CLD.

Birth Weight↗

Internal citrate ions reduce the membrane potential for contraction threshold in mammalian skeletal muscle fibers.

An effect of internal citrate ions on excitation-contraction coupling in skeletal muscle is described. The threshold for contraction was measured in rat extensor digitorum longus, (EDL), and soleus muscle fibers using a two microelectrode voltage clamp technique with either KCl-filled or K3 citrate-filled current electrodes. Contraction thresholds were stable for many minutes with KCl current electrodes. In contrast, thresholds fell progressively towards the resting membrane potential, by as much as -15 mV over a period of 10 to 20 min of voltage-clamp with citrate current electrodes. In addition, prepulse inhibition was suppressed, subthreshold activation enhanced and steady-state inactivation shifted to more negative potentials. Fibers recovered slowly from these effects when the citrate electrode was withdrawn and replaced with a KCl electrode. The changes in contraction threshold suggest that citrate ions act on the muscle activation system at an intracellular site, since the citrate permeability of the surface membrane is probably very low. An internal citrate concentration of 5 mM was calculated to result from citrate diffusion out of the microelectrode into the recording area for 20 min. 5 mM citrate added to an artificial cell lowered the free calcium concentration from 240 to 31 microM. It is suggested that citrate modifies excitation-contraction coupling either by acting upon an anion-dependent step in activation or by reducing the free calcium and/or free magnesium concentration in the myoplasm.

Animals↗

Citrate utilization by homo- and heterofermentative lactobacilli.

Citrate utilization by several homo- and heterofermentative lactobacilli was determined in Kempler and McKay and in calcium citrate media. The last medium with glucose permitted best to distinguish citrate-fermenting lactobacilli. Lactobacillus rhamnosus ATCC 11443, Lactobacillus zeae ATCC 15820 and Lactobacillus plantarum ATCC 8014 used citrate as sole energy source, whereas in the other strains, glucose and citrate were cometabolized. Some lactobacilli strains produced aroma compounds from citrate. Citrate transport experiments suggested that all strains studied presented a citrate transport system inducible by citrate. The levels of induction were variable between several strains. Dot blot experiment showed that lactobacilli do not present an equivalent plasmid coding for citrate permease.

Acetoin↗