Changes in plasma and urinary taurine and amino acids in runners immediately and 24 h after a marathon

Summary. Changes in urinary and plasma taurine and amino acids have been evaluated in trained runners competing in the Rotterdam Marathon, 1998, both immediately after completing the event and 24 h after recovery. There were significant changes in the urinary amino acids excretion, the majority showing a significant decrease both immediately at the completion of the Marathon and after 24 h recovery. In contrast urinary taurine excretion increased immediately post Marathon, although not significantly as the range of results was wide. Such changes in urinary taurine correlated with percentage changes in plasma creatine kinase both immediately post race, (r = 0.972, P < 0.001), and 24 h later (r = 0.872, P < 0.001), possibly indicating that the source of the taurine was muscle. Significant correlations between the individual values for urinary and plasma amino acids in all of the athletes were calculated for taurine (r = 0.528), glycine (r = 0.853), threonine (r = 0.749), alanine (r = 0.747), serine (r = 0.620), glutamine (0.614), arginine (r = 0.507), histidine (r = 0.470) and valine (r = 0.486). Changes in the mean plasma concentrations of amino acids were comparable to our previously published data (Ward et al., 1999) the majority showing significant decreases immediately and 24 h post Marathon, such an adaptation being due primarily to their utilisation for gluconeogenesis. However, in contrast, the mean taurine concentrations were significantly elevated both post race, P < 0.01 and after 24 h, P < 0.05. The physiological response by the muscle to exhaustive exercise, particularly with regard to changes in plasma and urinary taurine concentrations remain to be elucidated, but is probably related to muscle function impairment. The increase in taurine urinary excretion could be used as an indicator of muscle damage occurring during exhaustive exercise. Whether taurine supplementation would minimise such changes is an interesting scientific question and merits investigation. Received January 6, 2000 / Accepted February 1, 2000     

The biosynthesis of taurine fromN-acetyl-l-cysteine and other precursorsin vivo and in rat hepatocytes

Summary The synthesis of taurine fromN-acetylcysteine has been examined in ratsin vivo and in rat hepatocyte suspensionsin vitro. In ratsin vivo, administration ofN-acetylcysteine significantly increased urinary taurine (3 fold) 24h after dosing and liver glutathione levels. Liver taurine was not increased significantly. In hepatocytes incubated in the presence ofN-acetylcysteine, glutathione concentration increased to a maximum after 1 hour but the increase was not dependent on the concentration ofN-acetylcysteine. In contrast, after an initial lag phase, taurine synthesis increased in relation to the concentration ofN-acetylcysteine and continued for 3 hours. Glutathione synthesis seems to be preferential to taurine synthesis. Taurine synthesis from cysteine sulphinate was greater and from hypotaurine was greatest and maximal after 1 hour. Implications for the mechanism of protection byN-acetylcysteine are discussed.     

Inhibition of sulfate excretion by (aminooxy)acetate induced stimulation of taurine excretion in rats

Summary l-Cysteine is mainly metabolized to sulfate and taurine through cysteinesulfinate pathway. Alternatively, sulfate is formed in rat liver mitochondria via 3-mercaptopyruvate pathway. Intraperitoneal administration of 5 mmol ofl-cysteine per kg of body weight resulted in the increase in sulfate and taurine (plus hypotaurine) excretion in the 24-h urine, which corresponded to 45.3 and 29.3%, respectively, ofl-cysteine administered. Subcutaneous injection of (aminooxy)acetate, a potent inhibitor of transaminases, together withl-cysteine halved the sulfate excretion and doubled the taurine excretion. In vitro sulfate formation froml-cysteine and froml-cysteinesulfinate in rat liver mitochondria was inhibited by (aminooxy)-acetate. The sulfate-forming activity of liver mitochondria obtained from rats injected with (aminooxy) acetate was also inhibited. These results indicate that the transamination reaction is crucial in sulfate formation and in the regulation of sulfur metabolism. Sulfur equilibrium in mammals was discussed.     

Changes in plasma taurine levels after different endurance events

Summary The sulphonated amino acid taurine increased significantly in the plasma of trained athletes after three endurance exercises of different duration and intensity, a 90 min run on a treadmill at 75% of an individual's VO₂ peak, a Marathon, 42.2km and a 100km run, by 19%, 77% and 36%, respectively. Such results indicated that the speed at which the exercise is per formed, referred to as the intensity, rather than the duration of the exercise, correlated with the elevated taurine levels possibly indicating its release from muscle fibres. The plasma amino acid pool decreased significantly in relationship with the duration of the exercise, caused by their utilisation for glucogenesis. The possible sources of the increased plasma taurine are discussed.     

Production of hypotaurine, taurine and sulfate in rats and mice injected with L-cysteinesulfinate

Summary. We studied in vivo production of taurine, hypotaurine and sulfate following subcutaneous administration of L-cysteinesulfinate (CSA) to rats and mice. When 5.0 mmol/kg of body weight of CSA was injected to rats, increased urinary excretions of taurine, hypotaurine and sulfate in 24 h urine were 617, 52 and 1,767 μmol/kg, respectively. From these results together with our previous data, sulfate production was calculated to be 1.6 times greater than taurine production. Increased contents (μmol/g of wet tissue) over the control of taurine and hypotaurine in mouse tissues at 60 min after the injection of 5.0 mmol/kg body weight of CSA were: liver, 3.5 and 9.9; kidney, 0.3 and 5.2; heart, 3.7 and 0.2; blood plasma, 0.4 and 0.2, respectively. Upon loading of hypotaurine or taurine, tissue contents of these amino acids in liver and kidney increased greatly. Our results indicate that liver is the most active tissue for taurine production, followed by kidney, and that external CSA, hypotaurine and taurine are easily taken up by these tissues.     

The components of taurine transport across the rat small intestine A kinetic study

Summary The transport of taurine across adult Sprague-Dawley rat small intestine was studied in vitro using small intestinal strips. The kinetics of the transport mechanism were investigated under both steady-state and influx conditions. Our findings were compatible with the presence of two distinct transport mechanisms; a linear non-carrier mediated component and a saturable carrier mediated component, with almost equal contribution from each. The mediated component was found to be largely Na⁺-dependent and exhibited marked inhibition by B-alanine and structurally related sulfur amino acids.     

Physical and structural properties of taurine and taurine analogues

Summary In a variety of mammalian species it has been established that taurine is a necessary component of the visual system, however, the exact mechanism(s) as to the function of taurine is(are) elusive. Additionally, taurine is speculated to be a membrane stabilizer by interacting with phospholipids and a regulator of protein phosphorylation. Therefore the inhibition by taurine and taurine analogues of the phosphorylation of an 20 kDa protein present in the mitochondrial fraction of the rat retina has been investigated using computational methods. Correlations between molecular weight, molecular volume, and calculated pKa values vs. IC₅₀ values are reported. These data appear to support the hypotheses according to Lombardini and Props that the inhibition of the phosphorylation of an 20kDa protein by taurine and taurine analogues is dependent on (i) the critical distance between the nitrogen and sulfur atoms in the taurine moiety (S-C-C-N) of the analogue; (ii) the environment of the nitrogen atom in the taurine analogue (saturated ring vs. unsaturated ring); and (iii) the placement of both the sulfur and nitrogen atoms not being present simultaneously in the ring structure. Using computational methods we present results that support hypotheses (i) and (ii).     

Pharmacological characterization of the effects of taurine on calcium uptake in the rat retina

Summary Taurine is known to increase ATP-dependent calcium ion (Ca²⁺) uptake in retinal membrane preparations and in isolated rod outer segments (ROS) under low calcium conditions (10M) (Pasantes-Morales and Ordóñez, 1982; Lombardini, 1991). In this report, ATP-dependent Ca²⁺ uptake in retinal membrane preparations was found to be inhibited by 5M cadmium (Cd²⁺), suggesting the involvement of cation channel activation. The activation of cGMP-gated cation channels, which are found in the ROS, is a crucial step in the phototransduction process. An inhibitor of cGMP-gated channels, LY83583, was found to inhibit taurine-stimulated ATP-dependent Ca²⁺ uptake but had no effect on ATP-dependent Ca²⁺ uptake in the absence of taurine, indicating that taurine may be increasing ATP-dependent Ca²⁺ uptake through a mechanism of action involving the opening of cGMP-gated channels. The activation of cGMP-gated channels with dibutyryl-cGMP and with phosphodiesterase inhibition using zaprinast caused an increase in ATP-dependent Ca²⁺ uptake in isolated ROS, but not in taurine-stimulated ATP-dependent Ca²⁺ uptake. LY83583 had the same effects in isolated ROS as in retinal membrane preparations. Another inhibitor of cGMP-gated channels, Rp-8-Br-PET-cGMPS, produced the same pattern of inhibition in isolated ROS as LY83583. Thus, there appears to be a causal link between taurine and the activation of the cGMP-gated channels in the ROS under conditions of low calcium concentration, a connection that suggests an important role for taurine in the visual signalling function of the retina.     

Development of an LC-MS-MS method for the quantification of taurine derivatives in marine invertebrates

Sulfur amino acids, such as taurine, hypotaurine, and thiotaurine, were found in high quantities in tissues of marine symbiotic organisms (e.g., bivalves, tubeworms) living close to hydrothermal vent sites. Therefore, they are assumed to play a key role in the S-oxidizing base metabolism or sulfide detoxification. We propose here a specific, rapid, and original analytical procedure for the direct determination of sulfur amino acids at the level of a few parts per billion in biological samples, avoiding the classical low specific post-column ortho-phthaldialdehyde derivatization step required by non-ultraviolet-absorbing molecules. Indeed, by coupling liquid chromatography on a porous graphitic stationary phase under isocratic conditions (10 mM ammonium acetate buffer adjusted to pH 9.3) to tandem mass spectrometry (ionization process by pneumatically assisted electrospray in negative ion mode), it is possible to perform specific quantification of these metabolites in less than 10 min directly in biological matrices without any derivatization step or other tedious sample treatments. Thus, taurine, hypotaurine, and thiotaurine have been identified and assayed in several deep sea organisms, showing that the developed method is well suited for this kind of application.     

A study on the estimation of sulfur-containing amino acid metabolism by the determination of urinary sulfate and taurine

Summary.  Sulfate and taurine are major end products of sulfur-containing amino acid metabolism in mammals including humans, and they are excreted in urine. Average excretions (μmol/mg of creatinine) in the morning urine of 58 female college students were: total (free plus ester) sulfate (a), 12.53 ± 3.85; free sulfate, 11.57 ± 3.69; taurine, 0.78 ± 0.53. Ratio of total sulfate and taurine was 10 : 0.6. Regression lines obtained by plotting total sulfate, free sulfate, or total sulfate plus taurine against urea have shown that the former excretions are significantly correlated with urea excretion. Excretion of total sulfate at zero point of urea excretion (b) was 5.30, which corresponded to 42.3% of average excretion (12.53) and was assumed to be derived from dietary sulfate. The difference 7.23 (a − b) seemed to be derived from sulfur-containing amino acids. It was pointed out that the difference of average sulfate excretion and sulfate excretion at zero urea excretion, namely a − b, was appropriate for the metabolic index of sulfur-containing amino acids of the group examined. As free sulfate constituted 92.3% of total sulfate, excretion of ester sulfate was at a constant level, and that of taurine was not significantly correlated with urea excretion, the value of free sulfate corresponding to the value a − b of total sulfate mentioned above seemed to be a reliable and convenient index in the assessment of sulfur-containing amino acid metabolism. Received December 3, 2001 Accepted January 2, 2002 Published online August 30, 2002 Authors' address: Dr. Toshihiko Ubuka, Department of Clinical Nutrition, Kawasaki University of Medical Welfare, Kurashiki Okayama, 701-0193 Japan, E-mail: ubukatos@mw.kawasaki-m.ac.jp     

Kinetics of taurine depletion and repletion in plasma, serum, whole blood and skeletal muscle in cats

The relationship between taurine concentrations of plasma, whole blood, serum and skeletal muscle during taurine depletion and repletion was investigated in cats, to identify the most useful indicators of taurine status. Sixteen cats were fed a purified diet containing either 0 or 0.15 g/kg taurine for 5 months. Treatments were then reversed and the taurine concentration was measured during repletion and depletion phases. Plasma taurine exhibited the fastest rate (slow component) of depletion (t 1/2 = 4.8 wk), followed by serum (5.3 wk), whole blood (6.2 wk), and skeletal muscle (11.2 wk). Whole blood taurine was the first to replete at a rate of 0.74 wk to 1/2 maximal repletion, followed by serum (2.1 wk), skeletal muscle (3.5 wk), and plasma (3.5 wk). Whole blood more closely reflected skeletal muscle taurine concentrations than plasma during depletion, while plasma taurine concentrations appear to be the most valuable predictor of skeletal muscle taurine concentrations during repletion. This study suggests that the best clinical method to evaluate the taurine status of the cat is the determination and interpretation of both plasma and whole blood taurine concentrations.     

Role of taurine in osmoregulation in brain cells: Mechanisms and functional implications

Summary All cells including neurons and glial cells are able to keep their volume within a very limited range. The volume regulatory mechanism involves changes in the concentration of osmolytes of which taurine appears to be of particular importance in brain cells. Swelling in brain cells may occur as a result of depolarization or small fluctuations in osmolarity. In isolated brain cells these conditions will always lead to a release of taurine, the time course of which is superimposable on that of the volume regulatory decrease which follows the initial cell swelling. The mechanism responsible for taurine release associated with swelling has not been fully elucidated but a large body of evidence seems to exclude participation of the taurme high affinity carrier. Using a number of inhibitors of anion exchangers it has been demonstrated that both volume regulation and taurine release in brain cells are inhibited by these drugs, implicating an anion channel in the process. It has be controversial issue as to whether or not taurine release is Ca++ dependent. Recent evidence appears to suggest that the release process is not associated with Ca⁺⁺ or Ca⁺⁺ channels. It is, however, quite possible that the swelling process may involve the Ca⁺⁺ calmodulin system or other second messengers. Taurine also contributes to volume regulation after shrinkage of brain cells, in this case by increasing its intracellular concentration. This change is accomplished byan upregulation of the Na+/taurine cotransporter, together with reduced passive fluxes and increased endogenous synthesis.     

The influence of a taurine containing drink on cardiac parameters before and after exercise measured by echocardiography

Summary. To determine the effect of the taurine containing drink "Red Bull" on cardiac parameters thirteen endurance trained subjects performed an exhaustive bout of endurance exercise at three different times. Prior to the exercise the original "Red Bull" drink, a similar drink without taurine, containing caffeine, and a "placebo" drink without caffeine and without taurine were ingested by the subjects in a double-blind cross-over design. Echocardiographic examinations were performed before the drinks, 40 minutes after the drinks prior to the exercise and in the regeneration period after exercise. Stroke volume was significantly influenced only in the "Red Bull group" (80,4 ± 21,4 ml before drink vs. 97,5 ± 26,2 ml in the regeneration period), mainly due to a reduced endsystolic diameter and volume. Furthermore in this group the peak late diastolic inflow (V_A) in the regeneration period was significantly higher compared with the pre-exercise levels. This observation was also made in the caffeine group but without any consequences on ventricular function. The results of the present study show an influence of the original caffeine and taurine containing drink (Red Bull) on parameters of the cardiac contractility. Received September 9, 1999 / Accepted February 1, 2000     

Effects ofin vivo taurine depletion on induced- chemiluminescence production in macrophages isolated from rat lungs

Summary Alveolar macrophages isolated by pulmonary lavage from partially taurine-depleted ratsdemonstrated increased (2–2.6 fold) chemiluminescence due to the extracellular reaction between exogenous zymosan and various reactive forms of oxygen compared to macrophages isolated from control animals. Partial taurine depletion was achieved by adding 3% ß-alanine to the drinking water of the rats for 5 weeks prior to harvesting the macrophages. Superoxide dismutase activity was not increased in the lung tissue of the taurine-depleted rats. These data suggest that taurine has antioxidant properties and that taurine depletion is potentially deleterious to alveolar macrophages and pulmonary tissue.     

Identification of a specific protein in the mitochondrial fraction of the rat heart whose phosphorylation is inhibited by taurine

Summary It was previously reported that the mitochondrial fraction of the rat heart contained a specific protein with a molecular weight of approximately 44kDa whose phosphorylation was inhibited by taurine (Lombardini,1994a). Isolation of the 44kDa phosphoprotein on a 1-dimensional polyacrylamide gel using traditional glycine buffers followed by re-electrophoresing the cut out proportion of the gel which corresponds to the 44kDa protein on a tricine-buffered gel resulted in sufficient pure protein for sequence analysis. The results indicate that the 44kDa phosphoprotein is pyruvate dehydrogenase.     

Polarized nature of taurine transport in LLC-PK1 and MDCK cells: Further characterization of divergent transport models

Summary Taurine transport was measured in cultured epithelial cells-LLC-PK1 and MDCK-grown on permeable membrane supports. Taurine transport by LLC-PK1 cells was greater on the apical surface compared to the basolateral surface. MDCK cells exhibited greater taurine uptake from the basolateral side. Transepithelial taurine flux was in the direction of apical to basolateral in the LLC-PK1 monolayers. There was no net transepithelial movement of taurine in the MDCK monolayers. Efflux of taurine from the apical and the basolateral membrane surfaces of LLC-PK1 cell monolayers was stimulated by external-alanine but not L-alanine. Efflux of taurine from MDCK cell monolayers was stimulated by-alanine on the basolateral surface. While the competitive inhibitor guainidinoeithane sulfonate (GES) competitively inhibited taurine uptake to a similar degree on the apical and basolateral surface of LLC-PK1 cell monolayers, GES had a more potent inhibitory effect on the basolateral taurine uptake in MDCK cells when compared to its inhibition of apical taurine transport. We conclude that there are characteristic differences in transport of taurine by apical and basolateral surfaces of LLC-PK1 and MDCK cells which may be the consequence of asymmetric distribution or unique structural properties of the taurine transporter.Supported by a grant from the National Institutes of Health (DK 37223), the American Heart Association (92-004470).     

Effects of taurine and hypotaurine supplementation and ionophore concentrations on post-thaw acrosome reaction of dog spermatozoa

The aim of this work was to study of the effect of the amino acids (AA) taurine (T) and hypotaurine (H) and of different calcium ionophore concentrations on the ability of capacitated frozen-thawed dog sperm to undergo the acrosome reaction (AR). Fifteen ejaculates grouped into five pools were used. Sperm was frozen at a concentration of 80 × 10⁶ sperm cells/mL in the Uppsala Equex extender (UE) supplemented with 25, 50 and 75 mM of either AA. The UE extender without T or H was used as control. After thawing, sperm was capacitated with Canine Capacitation Medium for 20 min. Sperm was then challenged with calcium ionophore A23178 at 0, 2.5 and 10 μM concentration and evaluated for integrity of plasma and acrosome membranes after 5, 15 and 30 min of incubation, utilizing PI/Fitc-PNA fluorescent staining and flow cytometry.Sperm cryopreserved in UE supplemented with 50 mM T (UE 50T) had higher AR rates than sperm cryopreserved with UE 75T, UE 25H and UE 50H, but AR rates were similar to semen frozen with the control extender. Challenges with 2.5 and 10 μM/L of calcium ionophore increased AR in frozen-thawed sperm incubated for 5, 15 and 30 min. The combination of calcium ionophore concentration and incubation time resulting in the highest AR rate was 10 μM and 15 min.     

Chronic administration of valproic acid induces a decrease in rat striatal glutamate and taurine levels

Summary The effect of acute and chronic (10 days) administration of 200 mg/kg (i.p.) of valproic acid (VPA) on endogenous levels of aspartate, glutamate, alanine, glycine and taurine in the cerebral frontal cortex and corpus striatum of rats was studied. Quantification of the amino acid levels was performed by HPLC.Valproic acid (VPA) did not either induce changes on these neurotransmitters contents in corpus striatum after acute treatment. After chronic administration we found a decrease on the endogenous levels of glutamic acid (24%, p < 0.05) which was related to an increase (250%, p < 0.02) of the in vitro KCl evoked release of glutamate. We found decrements in taurine endogenous levels (22%, p < 0.05) which was not associated with an increase of its release.In cerebral frontal cortex there was not found any change neither under the acute nor under the chronic condition.Thus, it may be conclude that chronic treatment with VPA produces decreases on the endogenous levels of glutamate and taurine. However the relevance of this effect concerning it therapeutic action remains unclear.     

Shape and size changes induced by taurine depletion in neonatal cardiomyocytes

Summary Taurine is a very important organic osmolyte in most adult cells. Because of this property it has been proposed that large changes in the intracellular content of taurine can osmotically stress the cell, causing changes in its size and shape. This hypothesis was examined by measuring cell dimensions of taurine deficient cardiomyocytes using confocal microscopy. Incubation of isolated neonatal rat myocytes with medium containing 5mM-alanine led to a 55% decrease in intracellular taurine content. Associated with the loss of taurine was a reduction in cell size. Two factors contributed to the change in cell size. First, there was a shift in cell shape, favoring the smaller of the two cellular configurations commonly found in the myocyte cell culture. Second, the size of the polyhedral configuration was reduced after ßalanine treatment. These same two events also contributed to size reduction in cardiomyocytes incubated with medium containing 30mM mannitol. Nonetheless, some qualitative differences exist between cells osmotically stressed by increasing the osmolality of the incubation medium and decreasing intracellular osmolality. The results support a role for taurine in the regulation of osmotic balance in the neonatal cardiomyocyte.     

Antidiuretic hormone infusion reduces taurine and NaCl-induced hypernatremia in the rats

Summary Rats drinking taurine and hypertonic saline (T + S) develop severe hypernatremia, but rats drinking either T or S alone do not. One hypothesis for this disruption of homeostasis is that the T + S combination interferes with the actions of antidiuretic hormone (ADH). Rats drinking T + S developed severe hypernatremia (170 mmol/L) by day 8 when infused with distilled water by osmotic minipumps, but maintained plasma sodium below 150 mmol/ L when infused with ADH. Cumulative water balance in T + S drinkers receiving ADH was consistently higher than in those not receiving ADH. However the ratio of cumulative sodium balance to cumulative water balance suggests little uniform advantage to rats receiving ADH nor does comparison of urine osmolality in the two groups. Precisely how ADH administration reduces hypernatremia in T + S drinking rats remains unclear, but the hypothesis that T + S interferes with the action of ADH in its regulation of extracellular fluid volume and osmolality remains viable.Supported by FMHS Project Grant CP/96/22 and St. George's University.