Abstract: 1. The anti-inflammatory agent diclofenac sodium (o-[(2,6-dichlorophenyl)amino]phenylacetic acid sodium salt) is extensively metabolized by rat, dog, baboon and man. The main metabolites were isolated from the urine of all species and from the bile of rat and dog and identified by spectroscopy. 2. Metabolism involves direct conjugation of the unchanged drug, or oxidation of the aromatic rings usually followed by conjugation. Sites of oxidation are either position 3' or 4' of the dichlorophenyl ring or, alternatively, position 5 of the phenyl ring attached to the acetic acid moiety. 3. In the urine of rat, baboon and man conjugates of the hydroxylated metabolites predominate, but the major metabolite in dog urine is the taurine conjugate of unchanged diclofenac. 4. In the bile of rat and dog, the main metabolite is the ester glucuroniade of unchanged diclofenac.

Abstract: Taurine is one of those enigmatic substances about which so much, and so little, is known. It is a simple substance, with a molecular weight of 125 and a molecular formula of C2H7O3NS. Taurine occurs in high concentration in many mammalian tissues, comprising in excess of 60% of the total free amino acid pool in the rat heart. Furthermore, it is one of the most abundant amino acids in the brain. Taurine, which was first isolated from bull’s urine in 1827 (Tiedemann and Gmelin, 1827), is a substance of high chemical stability and low metabolic reactivity. In mammals, only one potential pathway for the degradation of taurine has been proposed, that being the conversion to isethionic acid, but the evidence for this pathway is tenuous (Huxtable, 1978). Analysis of taurine can be readily achieved by a number of simple colorimet-ric tests in the laboratory, or it can be detected by an amino acid analyzer, on which it is one of the first substances eluted.

Abstract: . Quantitative gas chromatography-mass spectrometry was used to study the metabolic profiles of unconjugated, conjugated and sulphated bile acids in urine of patients with intermittent intrahepatic cholestasis of unknown aetiology, cirrhosis of the liver, primary biliary cirrhosis, viral and toxic hepatitis and extrahepatic cholestasis. A large number of bile acids was present which can broadly be classified into four groups: cholic and chenodeoxycholic acids constituted between 49·4% and 77·9% of the total bile acids (mean values of the groups); deoxycholic and other 3,12-disubstituted bile acids between 1·3% and 12·3%, monohydroxy bile acids between 6·7% and 14·4% and bile acids hydroxylated at C-1 or C-6 between 4·6% and 14·6%. The high proportion of bile acids from the latter group, and the presence of tetrahydroxylated bile acids, clearly distinguished hepatic disease from the normal state. The metabolic profiles were very variable and there were few consistent differences between the groups of diseases studied. Norcholic acid constituted a significantly higher percentage of the total bile acids in cirrhotic patients (6·2 ± 6·8%) than in non-cirrhotic patients (1·3±1·8%, P<0·001). With this exception, no profile was specific for any type of intra- or extra-hepatic cholestasis. The excretion rates of the major l-hydroxylated bile acids were positively correlated to each other. The same was true for the major 6-hydroxylated bile acids. This may indicate that cholic, chenodeoxycholic and deoxycholic acids act as substrates for common 1- and 6-hydroxylating enzymes. Possibly the taurine conjugates are preferred substrates since 1-hydroxylated bile acids and hyocholic acid were found mainly in this fraction. A positive correlation between the excretion of sulphated 3β-hydroxy-5-cholenoic acid and 3β,12α-dihydroxy-5-cholenoic acid indicates a direct metabolic relationship between these compounds. Confirming previous data, a high proportion of bile acids was sulphated. The degree of sulphation increased with decreasing number of hydroxyl groups, reaching 100% for the monohydroxy and most of the dihydroxy acids. Tetrahydroxycholanoates were not sulphated, and sulphation of trihydroxycholanoates was positively correlated to the renal bile acid excretion rate. Bile from patients with intermittent intrahepatic cholestasis did not contain the tetrahydroxylated bile acids present in urine. Hyocholic acid was a very minor, mainly taurine conjugated, bile acid. Monohydroxy bile acids were usually below the detection limit. These data do not support the hypothesis that lithocholic acid participates in the initiation or perpetuation of intermittent intrahepatic cholestasis of unknown aetiology.

Abstract: — We surveyed the transport systems present in the brain for amino acids. Cellular transport was measured in brain slices, and capillary transport was estimated by measuring in vivo the short-term (15 s) extraction by brain from the blood. Specific analog inhibition of uptake was used to distinguish the classes. Amino acid levels (close to physiological) were such that primarily the ‘low-affinity’ transport was measured. In brain tissue we could distinguish 10 overlapping amino acid transport classes. Five of these, described in a number of tissues, were characterized by their substrates: alanine (A system), leucine (L system), alanine-serine-cysteine (ASC system), glutamic acid (Glu system), and arginine (Ly+ system), respectively. The others distinguished were each fairly specific for one of the following five amino acids: glycine, proline, γ-aminobutyric acid (GABA), taurine, and lysine. Of these 10 systems only 4 could be clearly found in capillary transport: L, ASC, Ly +, and Glu. The properties and the distribution of the transport systems are different. Examples are that at least one of the systems is present primarily only in neurons (GABA), and one primarily in glia (taurine). The specificity of some of the systems, e.g. A, is altered during development. In contrast to the properties of most other systems, there is little Na+, energy, or temperature dependence of the L system. This is reflected in the properties of capillary neutral amino acid transport when the L system is the predominant one.

Abstract: 1. Using a cortical cup technique, the release of seven endogenous amino acids from the isolated rat olfactory cortex slice has been monitored. 2. Electrical stimulation of the lateral olfactory tract at a frequency of 4 min−1 was accompanied by a significant increase in the release of aspartate, GABA and taurine; the release of GABA and aspartate but not that of taurine was Ca2+-dependent. 3. Chronic unilateral bulbectomy was accompanied by a specific, significant fall in the aspartate content of the olfactory cortex which reached a maximum 5 days after surgery and persisted for at least a further 5 days. Electrical stimulation of the lateral olfactory tract of such preparations did not release any of the amino acids under investigation. 4. When slices from the unoperated side were exposed to solutions containing 25 m M-K+ or stimulated with electrodes placed directly on the cortex enclosed by the cup there was a Ca2+-dependent release of aspartate and GABA accompanied by a Ca2+-independent release of taurine. Following chronic bulbectomy, these procedures failed to evoke significant release of aspartate whereas the characteristics of GABA and taurine release were unaltered. 5. It is concluded that aspartate may be the excitatory transmitter of some of the terminals of the lateral olfactory tract fibres and that GABA may be a transmitter at some inhibitory synapses of the rat olfactory cortex.

Abstract: It is known that hamster sperm require an unidentified low molecular weight “factor” found in several cells and tissue types in order to remain strongly motile during in vitro capacitation. The β-amino acid taurine (2-aminoethane sulfonic acid) is present in a partially purified “factor” from bovine adrenal glands and can substitute for that “factor” in maintaining and stimulating hamster sperm motility. Incubation of washed hamster sperm with bovine serum albumin and 2 × 10−3 to 2 × 10−5M taurine for periods of approximately 5 hr. resulted in a higher number of strongly motile sperm compared to controls in the absence of exogenous taurine. Incubation of sperm with (-)epinephrine in the absence of taurine resulted in nearly all sperm becoming immotile. Activation (whiplash flagellar movement characteristic of capacitated hamster sperm) did not occur in the absence of taurine, but high percentages of activation occurred in the presence of 2 × 10−3, 2−10−4 and 2 × 10−5M taurine. Acrosome reactions occurred in the presence of taurine, even in the absence of (-)-epinephrine. However, (-)-epinephrine, previously shown to stimulate activation and acrosome reactions, was required for the maximum number of the latter. These results suggest that in addition to its importance for motility taurine might stimulate capacitation. The mechanism through which taurine supports and stimulates motility is not yet known, but its use results in the first relatively “defined” in vitro capacitation medium for hamster sperm.

Abstract: The effect of excess methionine (MET) on cysteine (CYS) catabolism was investigated in rats prefed either a control (10% casein + 0.3% L-MET) or high-MET (10% casein + 3.0% L-MET) diet for 50 or 20 days. The activities of cysteine dioxygenase, cysteine desulfhydrase and cystathionase were increased in high-MET rats to levels 5.9, 2.7 and 2.7 times, respectively, those of control rats after 5 days and to 2.9, 2.0 and 2.7 times control levels after 20 days. Cysteine aminotransferase and cystathionine synthase activities were increased to 1.5 and 1.7 times control values after 5 days but were not significantly different from control values at 20 days. Following gastric intubation of 5 g of an L-amino acid diet containing 0.2% L-[35S]CYS, the 24-hour urinary exretion of 35SO4, [35S]taurine and total 35S (% of administered dose) and the [35S]taurine:35S and [35S]taurine:35SO4 ratios were increased in rats prefed excess MET for 5 or 20 days. When 2.6% L-[35S]CYS was administered similarly, no significant differences between high-MET and control rats were observed. However, the [35S]taurine:35S and [35S]taurine:35SO4 ratios were elevated in both high-MET and control rats given 2.6% L-[35S]CYS over those for control rats fed 0.2% L-[35S]CYS. The increase in cysteine dioxygenase activity and the increase in [35S]taurine:35SO4 ratio in rats fed excess MET or given a load dose of CYS suggest that the cysteine sulfinic acid pathway plays a major role in the regulation of CYS degradation.

Abstract: — With the single rat brain cortical slice serving as an in vitro bio-assay system, the effects of neurotransmitter amino acids (1 mm) on brain swelling, water, sodium and potassium content, inulin space, and lactate production were studied. The putative dicarboxylic amino acid neurotransmitters, l-glutamic acid and l-aspartic acids, greatly increased intracellular brain swelling with increased intracellular Na+, water content and lactate production, and decreased inulin space and intracellular K+. Equimolar GABA, taurine, glycine, the putative inhibitory neurotransmitter amino acids, and equimolar α-amino-isobutyric acid had no effect. Brain swelling and intracellular Na+/K+ ratios were greatly increased by l-glutamate and l-aspartate at a concentration of 10 mm. However, l-aspartate at these concentrations greatly depleted the K+ content and lactate production as compared to l-glutamate. Further studies indicated that only the structural analogs and isomers of the dicarboxylic amino acids possessing two acidic groups and an α-amino group had a similar effect on the induction of brain swelling. Among the analogs of glutamic acid, dl-homocysteic acid and kainic acid had a greater effect on brain swelling, as observed from the total adenosine 5′-triphosphate (ATP) levels and the time-course and dose-response. A biphasic response in lactate production was induced by dl-homocysteic acid and kainic acid, suggesting that these analogs had a neurotoxic effect on cellular metabolism at higher concentrations.

Abstract: Amino acid levels have been determined in plasma and in four cerebral regions of rats one month after portocaval shunt. Many plasma amino acids are significantly lowered (asparagine, glutamine, theonine, serine, alanine, valine, leucine, isoleucine, cystine, lysine), while others remain unchanged (taurine, glycine, proline, tryptophan, ornithine, histidine, arginine). Asparagine and glutamine levels are significantly higher than in normal rats, and a net increase of tyrosine (100%), phenylalanine (50%) and citrulline (50%) is evident. In the shunted rat brain the most prominent feature is a very large rise (up to fivefold) of tyrosine, phenylalanine, histidine, citrulline, tryptophan, and glutamine uniformly in the tested regions. Other neutral amino acids are slightly increased. Lysine and arginine are decreased in cerebellum and pons-medulla; taurine, in forebrain and cerebellum. Cerebral permeability to L-amino acids was studied in vivo. Neutral amino acid permeability is greatly increased, whereas basic amino acids show a net decrease in their rate of passage from blood to the brain. No changes are observed for GABA and glutamic acid. These data suggest an altered permeability of the cerebral capillary membranes, which seems to be selective for the different amino acid transport classes. Competitive inhibition experiments demonstrated that the increased brain permeability to neutral amino acids after portocaval shunt is due to an enhancement of the saturable transport. The sharp rise in the brain of some essential neutral amino acids (phenylalanine, tyrosine, trytophan, histidine), largely exceeding their changes in plasma, and the slight cerebral increase of other neutral amino acids despite their lowered level and the rise of competing amino acids in the plasma, is consistent with our observation of enhanced transport for the neutral class. In hepatic encephalopathy, correction of the altered plasma amino acid levels has been reported to improve the clinical status. If this result is connected to the concomitant correction of the brain amino acid levels, carefully selected competitive inhibition among various plasma amino acids could be a useful therapeutic tool in this pathologic condition. However, the increased activity of the neutral amino acid transport system adds a new factor to the problem, since it probably implies that the competing amino acids will accumulate to unphysiological levels in the brain.

Abstract: The quantitative and qualitative distribution of bile salts in the duodenal juice of 13 patients with cystic fibrosis (CF) was studied after a test meal. The effects of triolein (TO), bovine serum albumin (BSA), and ricinoleic acid (RA) on the absorption of taurocholate (TCA) in the distal ileum of the rat in vivo was also studied. The mean (and ranges) of total bile salt concentrations, glycine: taurine conjugate ratios, and percentage of dihydroxy bile salts in the patients with CF and pancreatic insufficiency were 3.5 (1.3--6.6) mmol/l, 8.6 (greater than 10-3.1), and 37 (10--60) compared with control values of 7.4 (3.0--16.0) mmol/l, 3.0 (1.3--4.5), and 61 (52--70) respectively. The differences between the control and CF values were statistically significant (P less than 0.01--P less than 0.001). Three of the 13 CF patients had total bile salt concentrations less than 2 mmol/l, 8 had much higher glycine: taurine ratios, and 8 had a reduced percentage of dihydroxy bile salts. In 2 patients with normal pancreatic enzyme activities, duodenal bile salts were both quantitatively and qualitatively normal. TO (10 and 30 mmol/l), BSA (3%), and RA (5 mmol/l) had no inhibitory effect on the ileal absorption of TCA. These results show pronounced abnormalities of duodenal juice bile salts in CF with pancreatic insufficiency consistent with a broken enterohepatic circulation (EHC); such abnormalities may contribute to defective lipid absorption in CF. The data in the experimental animal do not support the suggestion that unhydrolysed dietary substrates play a role in the pathophysiology of the broken EHC.

Abstract: — Crude synaptosomal (P2) preparations were obtained from the cerebella of rats in which the granule cell population had been selectively reduced by X-irradiation treatment and from the cerebella of control animals. In the P2 fraction from control cerebella, the level of glutamate was greater than any other of the 5 amino acids measured and was 2-fold higher than taurine, which was present at the next highest level. The content of taurine was slightly higher than that found for aspartate and was 3-fold greater than that observed for GABA. Alanine and glycine were present in the lowest amounts. The levels of glutamate and aspartate were significantly (P < 0.05) lower by 25 and 15%, respectively, in the P2 fraction isolated from the X-irradiated cerebella in comparison to control values. The content of taurine, GABA, glycine, and alanine were not changed by the X-irradiation treatment. The uptake of 1.0 μm-l-[3H]glutamate and l-[3H]aspartate was reduced approx 20% by X-irradiation treatment, whereas the uptake of 1.0 μm-[3H]GABA and [3H]taurine was unchanged. A more detailed kinetic analysis of l-[3H]glutamate uptake revealed there was a 20% decrease in the Vmax value with X-irradiation treatment and no change in the apparent Km value. In a second study, the uptake of l-[3H]glutamate, l-[3H]aspartate and [3H]GABA was measured using P2 fractions obtained from the cerebella of rats in which the population of granule, stellate and basket cells had been reduced by X-irradiation treatment. The uptake of 1.0μm-l-[3H]glutamate, l-[3H]aspartate and [3H]GABA was significantly (P < 0.05) reduced to 57, 68, and 59%, respectively, of control values. A more detailed kinetic analysis of [3H]GABA uptake revealed no significant change in the apparent Km and a 35% decrease in the Vmax value. The data are discussed in terms of glutamate being the excitatory neurotransmitter released from granule cells and GABA being the inhibitory neurotransmitter released from basket cells.

Abstract: The metabolic profiles of urinary bile acids in pregnant women in the last trimester and patients with recurrent intrahepatic cholestasis of pregnancy (RCP) were studied. Following separation according to mode of conjugation, about thirty different bile acids were quantitatively analysed by gas chromatography-mass spectrometry. In all patients the sulphate fraction comprised 50--90% of the total bile acids. In RCP a shift from glycine to taurine conjugation was noted to together with a slight relative increase in sulphation. A ten- to hundred-fold increase in cholic and chenodeoxycholic acids was seen in RCP, the increase being mainly in the sulphate fraction. Tetrahydroxylated bile acids, tentatively regarded as 1- and 6-hydroxylated products of cholic acid, were quantitatively important in patients with RCP. The relative amounts of the secondary bile acids, deoxycholic and lithocholic acids, decreased with increasing cholestasis. Metabolites hydroxylated at C-6 were common, and the excretion of hydroxylated at C-6 were common, and the excretion of hyocholic acid was positively correlated to that of chenodeoxycholic acid. An increase in the excretion of 5 alpha-configurated bile acids in RCP was noted. A positive correlation between the excretion of 3 beta-hydroxy-5-cholenoic acid and 3 beta,12 alpha-dihydroxy-5-cholenoic acid indicates a metabolic relationship between the two compounds. Because of the relatively small amounts of lithocholic and 3 beta-hydroxy-5-cholenoic acids in patients with RCP, these compounds do not seem to be of pathogenetic importance in this type of cholestasis.

Abstract: A severe disorganization of the lattice arrangement of tapetal rods occurs in cats nutritionally deprived of taurine. This is the first reported example of a nutritionally related degeneration of tapetal cells. The tapetal cell degeneration, along with the previously noted photoreceptor degeneration in taurine-depleted cats, suggests that taurine plays a vital role in maintaining the structural integrity of some biological structures. The visual evoked potentials were severely reduced in taurine-depleted cats as were the electroretinogram responses, as previously noted, indicating that they are deficient in light processing. These results also emphasize the importance of dietary taurine for the maintenance of normal structure and function of the eye, at least in the cat.

Abstract: Net taurine transport across the frog retinal pigment epithelium-choroid was measured as a function of extracellular potassium concentration, [K+]o. The net rate of retina-to-choroid transport increased monotonically as [K+]o increased from 0.2 mM to 2 mM on the apical (neural retinal) side of the tissue. No further increase was observed when [k+]o was elevated to 5 mM. The [K+]o changes that modulate taurine transport approximate the light-induced [K+]o changes that occur in the extracellular space separating the photoreceptors and the apical membrane of the pigment epithelium. The taurine-potassium interaction was studied by using rubidium as a substitute for potassium and measuring active rubidium transport as a function of extracellular taurine concentration. An increase in apical taurine concentration, from 0.2 mM to 2 mM, produced a threefold increase in active rubidium transport, retina to choroid. Net taurine transport can also be altered by relatively large, 55 mM, changes in [Na+]o. Apical ouabain, 10(-4) M, inhibited active taurine, rubidium, and potassium transport; in the case of taurine, this inhibition is most likely due to a decrease in the sodium electrochemical gradient. In sum, these results suggest that the apical membrane contains a taurine, sodium co-transport mechanism whose rate is modulated, indirectly, through the sodium pump. This pump has previously been shown to be electrogenic and located on the apical membrane, and its rate is modulated, indirectly, by the taurine co-transport mechanism.

Abstract: 1. The influx of [3H]gamma-aminobutyric acid ([3H]GABA) into isolated rat superior cervical ganglia has been measured by radioassay, supplemented by autoradiography. Ganglia were incubated in oxygenated Krebs solution at 25 degrees C, containing 10 microM-amino-oxyacetic acid. Under these conditions more than 95% of accumulated tritium was unmetabolized [3H]GABA. 2. Ganglionic radioactivity increased linearly with incubation time, to yield an intracellular fluid/extracellular fluid concentration ratio (Ci/Co) of about 200 after 6 hr in 0.5 microM-external [3H]GABA. 3. Uptake showed saturation with an apparent transport constant (KT) of 6.8 microM and maximum influx velocity (Jmaxi) of 7 mumole 1. cell fluid-1- min-1. 4. The influx rate at Co = 0.5 microM was unaltered by raising intracellular GABA from 0.2 to 1 mM. 5. Influx velocity increased with temperature (5--35 degrees C) in a monotonic manner with an apparent activation energy of 14 kcal mole-1. 6. Concentrative uptake was depressed by reducing external [Na+] with ouabain, by raising [K+]o above 20 mM, or by removing external Cl-. Uptake was not particularly sensitive to Ca2+ or Mg2+ ions. 7. Utake of [3H]GABA (0.5 microM) was inhibited by beta-guanidinopropionic acid (apparent KI, 28 microM), beta-alanine (KI, 55 microM), gamma-amino-beta-hydroxybutyric acid (KI, 220 microM), beta-amino-n-butyric acid (KI, 708 microM), 3-aminopropanesulphonic acid (KI, 832 microM) and taurine (KI greater than 1 mM). Uptake was not depressed by 1 mM-glycine, alpha-alanine, leucine, serine, methionine or alpha-amino-iso-butyric acid. 8. Radioactively labelled methionine, leucine, glycine, serine, beta-alanine and taurine (concentrations less than or equal to 5 microM) were also taken up by ganglia. Of these, only uptake of beta-alanine and taurine were significantly depressed by 1 mM-GABA. 9. Autoradiographs confirmed that [3H]GABA and [3H] beta-alanine were taken up predominantly into extraneuronal sites (presumed to be neuroglial cells). Methionine, leucine, glycine and serine showed preferential accumulation in neurones. Neuronal uptake of leucine was not prevented by inhibiting protein synthesis. 10. Calculations of net fluxes from unidirectional tracer fluxes suggest that the sympathetic glial cells are capable of promoting net uptake of GABA at external concentrations above 1 microM.

Abstract: The uterine uptake of amino acids was studied in 10 pregnant sheep with gestational ages of 114-146 days. After recovery from surgery, arterial and uterine venous samples were drawn simultaneously via indwelling catheters and analysed for amino acid and oxygen content. In seven ewes, amino acid concentrations were measured by a chromatographic technique. In four ewes, glutamate and glutamine arterio-venous differences across the uterine and umbilical circulations were measured by an enzymatic method. The uptake of neutral and basic amino acids was 66 mumol/mmol O2 and 17.3 mumol/mmol O2, respectively. Comparison of uterine and umbilical uptake shows that the bulk of the neutral and basic amino acids taken up by the pregnant uterus are transferred to the fetus. there was no significant uptake of acidic amino acids (i.e. glutamate, aspartate and taurine). glutamate was delivered from the fetus to the placenta but excretion of glutamate into the uterine circulation was negligible. Glutamine and asparagine were delivered to the fetus in amount which were two to three times larger than the placental uptake of glutamate and aspartate. Therefore placental conversion of exogenous glutamate and aspartate to glutamine and asparagine cannot account entirely for the fetal uptake of these amino acids.

Abstract: The free amino acids in eccrine sweat collected from the forearms of 20 healthy trained and 20 healthy untrained men during controlled exercise were determined quantitatively using ion exchange column chromatography. Sweat was deproteinized by adding an equal volume of 5% sulphosalicylic acid. The amino acid concentrations showed a constant qualitative pattern in sweat and large individual differences. Essential amino acids, such as isoleucine, leucine, lysine, methionine, phenylalanine, and valine were excreted in relatively small amounts. As compared to the trained men, untrained men showed statistically significantly higher concentrations in sweat for the following amino acids: Alanine, arginine, glycine, histidine, isoleucine, leucine, lysine, ornithine, phenylalanine, serine, taurine, threonine, tyrosine, and valine. No significant differences were found for citrulline, cystine, ethanolamine, and methionine. The comparison of the amino acid excretions in sweat obtained under controlled exercise and in urine showed that the amounts of amino acids excreted in sweat under controlled exercise were comparable to the losses of amino acids in urine.

Abstract: Taurine resembles G ABA in its synaptic effects in the cockroach cereal nerve giantfiber synapse where it exerts a depressant action upon synaptic transmission. Both taurine and G A B A produce an increased conductance of pre- and postsynaptic membranes through changes in the permeability of chloride ions.

Abstract: Uptake of [35S]taurine was studied in parallel on glial and neuronal cells maintained in continuous culture, including transformed neuronal cells. Both glial and neuronal taurine uptake systems were concentrative, highly sodium-dependent and inhibited by closely related structural analogues such as hypotaurine, β-alanine and GABA. Strychnine was found to be a potent inhibitor of taurine uptake, especially in the glial cells, while parachloromercuriphenylsulphonate was more efficient on the neuronal clones. In contrast with uptake by neuroblastoma cells, the glial transport was dependent on the presence of calcium in the incubation medium.