Abstract: Excitatory amino acids have been implicated in the production of calcium mediated neuronal death following central nervous system ischemia. We have used microdialysis to investigate changes in the extracellular concentrations of amino acids in the spinal cord after aortic occlusion in the rabbit. Glutamate, aspartate, glutamine, asparagine, glycine, taurine, valine, and leucine were measured in the micordialysis perfusate by high pressure liquid chromatography. The concentrations of glutamate, glycine, and taurine were significantly higher during ischemia and reperfusion than controls. Delayed elevations in the concentrations of asparagine and valine were also detected. The elevation of glutamate is consistent with the hypothesis that excitotoxins may mediate neuronal damage in the ischemic spinal cord. Increased extracellular concentrations of asparagine and valine may reflect preferential use of amino acids for energy metabolism under ischemic conditions. The significance of increased concentrations of inhibitory amino acid neurotransmitters is unclear.

Abstract: Our results show that a lack of taurine in the diet of cats results in a significant leukopenia, a shift in the percentage of polymorphonuclear and mononuclear leukocytes, an increase in the absolute count of mononuclear leukocytes, and a change in the sedimentation characteristics of white cells. Functional studies of polymorphonuclear cells isolated from cats fed taurine-free diets show a significant decrease In the respiratory burst as measured by chemliuminescence as well as a decrease in phagocytosis of Staphylococcus epidermis compared to cats fed the same diet containing taurine. In additIon, serum gamma giobulin in cats fed taurine-free diets was significantly increased compared to taurine-suppiemented cats, indicating that other immune cells may be affected by taurine deficiency. Histological examination of lymph nodes and spleen revealed regression of folllcular centers with depletion of reticular cells, mature and Immature lymphocytes (B cell areas), as well as mild extravascular hemolysis. These results indicate that there are profound immunologic consequences in cats with prolonged taurlne deficiency.

Abstract: The aim of the present study was to investigate the effects of 5-aminosalicyclic acid (5-ASA) on the cell injury mediated by activated neutrophils. We used a system constituted of neutrophils, triggered with phorbol myristate acetate, and 51Cr-labelled Daudi cells as targets. The results show that 5-ASA is capable of efficiently preventing neutrophil-mediated lysis. 5-ASA was up to 10-fold more effective than taurine, which acts as an hypochlorous acid scavenger. Moreover, 5-ASA was found to compete with taurine for the neutrophil derived hypochlorous acid. The results are consistent with the conclusion that 5-ASA is capable of limiting the neutrophil mediated cell damage by scavenging the generated hypochlorous acid. This may represent a potential mechanism for the therapeutic action of 5-ASA in ulcerative colitis.

Abstract: 1 Caffeine (10 mM) induced a transient contracture in saponin-treated cardiac trabeculae as a result of Ca2+ release from the sarcoplasmic reticulum (SR) Regular cycles of uptake and release were repeated to stabilize responses The SR accumulated Ca2+ during the period prior to the addition of caffeine and this was reflected in the size of the caffeine contracture Increasing the time for Ca2+ loading between successive caffeine exposures resulted in an increase in the amplitude of the contracture Similarly, the size of the contracture was a function of the calcium ion concentration [( Ca2+]) in the preceding loading period 2 Taurine (microM-40-mM), when included in both loading and caffeine solutions, markedly potentiated the caffeine-induced contracture The effect occurs even if taurine was included only in the loading solutions The potentiating effect was ascribed to a direct action of taurine on the SR, since taurine did not significantly change the [Ca2+] in the loading solutions 3 The maximal effect of taurine was produced at approximately 5 mM; higher taurine concentrations caused a lesser potentiation of the caffeine contracture If the solutions were balanced with respect to osmolarity the effect of taurine did not decline at high concentrations 4 If the [Ca2+] in the loading solutions was increased to produce a caffeine-induced contracture that peaked close to maximal Ca2(+)-activated force, taurine caused a fall in the size of contracture and a more variable response This result could be explained by an increase in the spontaneous release of Ca2+ from the SR in the presence of taurine 5 In Triton-skinned trabeculae, taurine (1 mM-40 mM) increased the Ca2+ sensitivity of the contractile proteins in a dose-dependent manner but had little effect on maximum Ca2(+)-activated force The increase in Ca2+ sensitivity was small: in a typical experiment 30 mM-taurine reduced the [Ca2+] necessary for half-maximal activation from 302 to 256 microM, with no significant change in the shape of the relationship

Abstract: Effects of increased concentrations of potassium and of hyposmolar conditions on release of taurine were investigated in cerebellar granule neurons cultured from mice. It was found that increases in the external potassium concentration as well as decreases in osmolarity dose-dependently increased release of exogenously supplied [3H]-taurine and endogenous taurine from the neurons. The release of endogenous taurine elicited by a reduction of the osmolarity of the incubation media to 70% or 50% was much more pronounced than that of other amino acids, particularly glutamine, the release of which was not affected at all. The potassium-stimulated release of [3H]-taurine was strictly chloride dependent and it was inhibited by an increase of the osmolarity of the media as well as by 4,4'-diisothyocyanostilbene-2,2'-disulfonate (DIDS) (100 microM). Moreover, a similar increase in the potassium concentration led to an increase in intracellular volume (swelling), a process which was also chloride dependent. It is concluded that potassium-stimulated taurine release from cerebellar granule neurons is associated with cell volume changes and that taurine is likely to play a role as an osmotically active substance in these neurons.

Abstract: The placenta absorbs anionic amino acids from the maternal and fetal circulations but does not significantly transfer these amino acids from mother to fetus. Uptake of L-aspartate and L-glutamate by basal (fetal-facing) plasma membrane vesicles from placental syncytiotrophoblast was stimulated by an inward sodium and an outward potassium gradient. Measurable saturable uptake was entirely sodium dependent and electrogenic. Studies of concentration dependence resolved a high-affinity (microM) system that has characteristics of the X-AG system found in other tissues including the placental microvillous plasma membrane. Uptake of 0.2 microM L-glutamate was inhibited by 2 mM L-glutamate, L-aspartate, D-aspartate, L-cysteate, and L-cysteinesulfinic acid and was uninhibited by 2 mM D-glutamate, L-glutamine, L-alanine, L-serine, L-asparagine, and taurine or by 1 mM methylaminoisobutyric acid. The X-AG system in the two membranes of the placental syncytiotrophoblast may mediate the concentrative uptake of anionic amino acids from the maternal and fetal circulations into the placenta.

Abstract: The effects of [K+]o on taurine release from glial cells were studied with primary cultures of cerebellar astrocytes and with LRM55 cells, a continuous glial cell line. The characteristics of K(+)-stimulated taurine release were virtually identical in the 2 cell types. Both cerebellar astrocytes and LRM55 cells released taurine when stimulated with high-K+ medium prepared by isosmotically substituting KCl for NaCl, but neither cell type released taurine when stimulated with hyperosmotic high-K+ medium prepared by adding solid KCl to control medium. The membrane potential of LRM55 cells was measured by intracellular recording and was insensitive to changes in [K+]o below 20 mM. LRM55 cells released taurine when stimulated with nondepolarizing concentrations of K+ (13–22 mM) if the isosmotically prepared high-K+ medium was used, but the cells did not release taurine when treated with a depolarizing concentration of K+ (50 mM) if hyperosmotic high-K+ medium was used. The time course of K(+)- stimulated taurine release was quite slow, having a time to peak of 10– 15 min. Small changes (2.5–10%) in the osmolarity of the medium strongly affected taurine release by cerebellar astrocytes and LRM55 cells. K(+)-stimulated taurine release from both cell types was inhibited when the osmolarity was increased with sucrose or NaCl and was enhanced when the osmolarity was reduced. Similarly, baseline taurine release was suppressed by small elevations in osmolarity and increased by reduced osmolarity.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: We examined the cytoprotective action of individual amino acids in isolated perfused kidneys during perfusion with either 10 mM lactate or 5 mM glucose. In the absence of amino acids inulin clearance fell rapidly, whereas fractional excretion of phosphate, lactate, or glucose increased to more than 30%; lactate dehydrogenase was released into perfusate and alkaline phosphatase into the urine. Functional deterioration was less in kidneys from rats rendered chronically water diuretic by drinking 5% glucose. Adding 5 mM glycine, L-alanine, beta-alanine, or D-alanine to the perfusate also prevented functional deterioration and release of enzymes. Glycine perfusion increased total phospholipid per microgram DNA by 6%. Aspartate, glutamate, glutamine, taurine, isoleucine, leucine, and valine were not protective. Serine, proline, and alpha-aminoisobutyric acid had small protective effects. Micropuncture measurements of proximal tubular free- and stop-flow pressures showed no effect of L-alanine on glomerular hemodynamics. L-Alanine increased oxygen consumption by both glucose- and lactate-perfused kidneys and increased gluconeogenesis by lactate-perfused kidneys but did not alter renal ATP content or energy charge. L-Alanine was not consumed during 70 min of perfusion and its protective action was not inhibited by blocking transamination with 0.5 mM amino-oxyacetate. The protective action of glycine was not inhibited by blocking glycine metabolism with 0.1 mM cysteamine. Thus the beneficial effects of L-alanine and glycine do not require their metabolism. These observations suggest that small neutral amino acids prevent tubular disruption through their physicochemical effects, which can stabilize membrane protein tertiary structure.

Abstract: It has been suggested that sugar cataracts associated with diabetes mellitus result from the accumulation of excess sorbitol within lens fibrils. Swelling of lens fibrils occurs when water moves in to maintain osmotic balance; the excess water causes disruption of fibrils and cataract formation. Other studies have indicated that more than sorbitol-induced osmotic stress is involved. Our study used lenses collected from rats after 21 or 44 d of streptozotocin diabetes. Cataracts formed in untreated 44-d streptozotocin diabetic rats, but were not apparent in the 21-d untreated diabetic animals. Lens sorbitol increased in the diabetic animals both before and after cataract formation. Lens taurine varied inversely with the sorbitol content in a fashion that resulted in no net change in total lens osmoles. Lens water did not increase in the diabetic animals with or without cataracts. The aldose reductase inhibitor Sorbinil prevented the increase in lens sorbitol in both the 21- and 44-d streptozotocin diabetic rats; cataract formation was prevented in the 44-d diabetic animals. The lens water in untreated diabetic animals with cataracts did not differ from lens water in the Sorbinil-treated diabetic animals that did not develop cataracts. Sorbinil treatment of diabetic animals was associated with normalization of both lens sorbitol and taurine levels. Taurine has been shown to serve both as an osmoregulator and as an antioxidant. The apparent increase in lens osmolality attributed to sorbitol was counterbalanced by an equimolar reduction in taurine concentration. The reciprocal relationship between taurine and sorbitol reduces the likelihood of an osmotic mechanism for sugar cataractogenesis; the reduced lens taurine, however, may increase the risk of lens protein oxidation and subsequent cataract formation. Thus in vivo sugar cataract formation may be an oxidative process rather than an osmotic phenomenon.

Abstract: The urinary excretion of taurine by rats after dosing with various hepatotoxins has been investigated by1H NMR spectroscopy. After single hepatotoxic doses of hydrazine, carbon tetrachloride, 1-naphthylisothiocyanate, or thioacetamide there was biochemical and histopathological evidence of hepatic damage. Proton NMR spectroscopy of the urine collected for 24 h after dosing from these animals revealed a marked elevation in taurine (control 11.9 μmole/h/kg) after dosing with thioacetamide (42.2 μmole/h/kg), carbon tetrachloride (52.5 gmmole/ h/kg), 1-naphthyl-isothiocyanate (80.4 μmole/h/kg) and hydrazine (52.9 μmmole/h/kg). After allyl alcohol administration there was no increase in taurine excretion (7.5 μmol/h/kg). The excretion of taurine after hydrazine administration was dose related. High resolution proton NMR spectroscopic analysis of urine also revealed resonances from several metabolites of hydrazine, an N-acetylcysteine conjugate of allyl alcohol, and acetamide as a metabolite of thioacetamide after dosing with the respective compounds. Changes in endogenous substances that may be related to the pathological events were also detected, such as a decrease in the excretion of 2-oxoglutarate and citrate after both hydrazine and carbon tetrachloride administration. The results confirm that proton NMR spectroscopic analysis of urine is a powerful analytical tool for the evaluation and study of toxic substances. Furthermore, measurement of urinary taurine may provide a non-invasive indicator of acute hepatic damage with certain classes of hepatotoxins.

Abstract: Human placental uptake and maternal-to-fetal (M-to-F) net transfer of taurine were evaluated in purified microvillous membrane vesicles (MMV) and the isolated perfused cotyledon Taurine uptake by MMV was specifically stimulated by an inward Na+ gradient [maximum velocity (Vmax), 245 +/- 06 pmolmg-130 s-1; Michaelis constant (Km), 62 +/- 07 microM], with uptake stoichiometry showing approximately 2 Na+/taurine molecule In the presence or absence of Na+, Cl- did not stimulate uptake Na(+)-stimulated uptake was enhanced by an outward K+ gradient beta-Alanine and hypotaurine competitively inhibited uptake of taurine in MMV Two-way uptake inhibition studies showed no interaction between taurine and non-beta-amino acids When MMV were preloaded with taurine there was enhancement of Na(+)-stimulated uptake In the perfused placentas, saturable M-to-F net transfer of taurine was not observed, despite saturation of tissue uptake from the maternal circulation During 3 h of perfusion, no fetal-to-maternal (F-to-M) gradient formed for taurine; yet, a 2:1 gradient simultaneously occurred for histidine This study demonstrates that taurine uptake by the microvillous membrane of the human placenta is highly specific, of high affinity, and Na+ coupled

Abstract: It is well established that excitatory amino acid neurotransmitters are extensively liberated during ischemia and that they have neurotoxic properties contributing to neuronal injury. To study changes in the liberation of excitatory and other amino acids during cerebral ischemia, we measured their extracellular concentrations and related them to blood flow levels and electrophysiologic activity (electrocorticogram and auditory evoked potentials) before and for up to 2 hours after multiple cerebral vessel occlusion in 14 anesthetized cats. Blood flow levels between 0 and 43 ml/100 g/min were reached. Concentrations of the excitatory amino acid neurotransmitters increased most (aspartate 10-fold, glutamate 30-fold, and gamma-aminobutyric acid 300-fold compared with control values) below a blood flow threshold of 20 ml/100 g/min. The total power of the electrocorticogram and the amplitude of the auditory evoked potentials were affected below the same blood flow threshold. In contrast, concentrations of the nontransmitter amino acids taurine, alanine, asparagine, serine, and glutamine increased 1.5-5-fold as blood flow decreased, while concentrations of the essential amino acids phenylalanine, valine, leucine, and isoleucine did not change during cerebral ischemia. The great increases in concentrations of the excitatory amino acid neurotransmitters below a blood flow threshold close to that for functional disturbance is in accordance with the role of these amino acids in ischemic cell damage. Their release at blood flow levels compatible with cell survival and the increase in their concentrations with severity and duration of cerebral ischemia imply that excitotoxic antagonists may have potential as therapeutic agents.

Abstract: Both cysteinesulfinate-independent and cysteinesulfinate-dependent pathways are involved in the catabolism of cyst(e)ine by freshly isolated rat renal cortical tubules. Sulfate and thiosulfate were shown to be the major sulfur-containing products that accumulated in incubations of renal tubules with 1 mmol/L or 25 mmol/L [35S]cyst(e)ine. Thiosulfate is an intermediate in the oxidation of the sulfide produced by the cysteinesulfinate-independent catabolism of cyst(e)ine by desulfhydration pathway(s), whereas sulfate is formed both by further oxidation of thiosulfate and by oxidation of the sulfite formed by the cysteinesulfinate-transamination pathway. Incubation of renal tubules with propargylglycine inhibited gamma-cystathionase activity by 85%, and this resulted in a 46% decrease in sulfate production and a 68% decrease in thiosulfate production when the treated renal tubules were incubated with 1 mmol/L [35S]cyst(e)ine. Addition of 25 mmol/L unlabeled cysteinesulfinate to create a diluting/trapping pool for [35S]cysteinesulfinate formed from [35S]cysteine resulted in a 53% decrease in [35S]sulfate production in incubations with 1 mmol/L cysteine. Thus, some cyst(e)ine catabolism probably occurred by a cysteinesulfinate-dependent pathway. No production of taurine or hypotaurine was detected in incubations with cyst(e)ine. Thus, cysteinesulfinate formed from cysteine was further catabolized primarily to sulfate instead of to taurine and hypotaurine. Most cyst(e)ine catabolism by the epithelial cells of the renal tubule probably can be accounted for by two pathways: 1) the beta-cleavage of cystine catalyzed by lambda-cystathionase and 2) the formation and transamination of cysteinesulfinate catalyzed by cysteine dioxygenase and aspartate aminotransferase.

Abstract: The effects of the naturally occurring amino acid taurine (2-aminoethanesulphonic acid) on isometric force development were investigated using skinned muscle fibre preparations. In atrial and ventricular pig heart muscles, as well as in fibres of slow abdominal extensor muscle of crayfish, an increase of submaximal isometric force was observed in Ca2(+)-activated skinned fibre preparations at physiological concentrations of taurine. The maximal isometric force remained unaffected in all preparations. It is assumed that taurine increases the Ca2+ sensitivity of the force-generating myofilaments in mammalian hearts and crustacean slow skeletal muscle fibres.

Abstract: The uptake and metabolism of 35S-labelled sulphur amino acids were compared in periportal (PP) and perivenous (PV) rat hepatocytes, isolated by digitonin/collagenase perfusion, to identify the factors underlying the previously observed [Kera, Penttila & Lindros, Biochem. J. (1988) 254, 411-417] higher rate of GSH replenishment in PP cells. The buthionine sulphoximine-inhibitable synthesis of GSH was faster in PP than in PV hepatocytes with both cysteine (6.1 versus 5.0 mumol/h per g of cells) and methionine (4.5 versus 3.3 mumol/h per g) as well as with endogenous precursors and L-2-oxo-4-thiazolidinecarboxylate as substrates. However, the uptake of cysteine by PP cells was slower than by PV cells (8.6 versus 10.3 mumol/h per g of cells), whereas methionine was taken up at similar rates. The activity of gamma-glutamylcysteine synthetase (GCS) was slightly higher in digitonin lysates from the PP than from the PV zone. Production of sulphate, the major catabolite of [35S]cysteine sulphur, as well as incorporation of the label into protein occurred at similar rates in PP and PV cells. Taurine, on the other hand, was produced from [35S]cysteine much faster by PV than by PP cells (0.7 versus 0.1 mumol/h per g of cells). Accordingly, the taurine content of PV hepatocytes tended to be higher and to increase faster during incubation with methionine. These results imply that metabolism of taurine is highly zonated within the acinus. They also suggest that both the slightly lower GCS activity and the fast metabolism of cysteine to taurine limit the capacity of PV hepatocytes to synthesize GSH.

Abstract: Merino sheep were given abomasal infusions of various amino acids or mixtures of amino acids. Effects on wool growth were measured using autoradiography or a clipping procedure and changes in the concentration of amino acids in plasma were measured in some experiments.Mixtures of five (28 g/day) or ten (45 g/day) essential amino acids (both mixtures containing 3 g methionine) stimulated wool growth of sheep receiving a maintenance ration; on average, the volume of wool grown increased 48% and 86%, respectively. When cysteine completely replaced methionine in these mixtures, wool growth was markedly reduced, but two-thirds of the methionine could be replaced by cysteine without affecting wool growth. Homocysteine was partially effective in replacing methionine and, when supplemented with betaine, folic acid and vitamin B12, the mixture was still significantly inferior to that containing methionine. In contrast, abomasal supplements of methionine or homocysteine alone were equivalent as supplements for wool growth. The results indicated a specific role for methionine in the control of wool growth, other than the provision of cysteine. This role was postulated to be related to some function of S-adenosylmethionine.Infusion often essential amino acids caused appreciable increases in the concentrations of cystine, methionine, cystathionine and taurine in plasma; total essential amino acids increased threefold whereas nonessential amino acids decreased in concentration. The replacement of methionine in the infusion by cysteine or homocysteine significantly altered the concentration of cystine, methionine and cystathionine in plasma.Evidence was obtained that the adverse effects on wool growth of high abomasal doses of methionine (10g/day) could not be reduced or prevented by provision of additional glycine and were not related to the supposed toxic effects of 3-methylthiopropionic acid, a metabolite of the transamination pathway.

Abstract: The β-sulphonic amino acid taurine is synthe-sised in animals from dietary sulphur amino acids. Cats exclusively use taurine to conjugate cholic acid rather than being able to use the alternate glycine conjugation. Since total body synthesis of taurine in cats is limiting, metabolic deficiencies of taurine occur when the dietary intake of taurine is restricted. A deficiency of taurine in cats is expressed by aberrant functions of a wide range of organ systems. Pathological changes occur in the eye, feline central degeneration; reproductive abnormalities occur in the female, a high incidence of fetal resorptions and abortions, low birth weight and survival of live-born young; growth rate in the new born kitten is depressed; heart induction of dilated cardiomyopathy and compromised immune function. All these conditions are prevented or reversed with adequate dietary taurine. While all tissues contain taurine, the concentration varies with the tissue. Generally plasma has been used to assess taurine status in cats, but the concentration of taurine in plasma varies widely. Food deprivation of cats given high taurine diets causes a marked fall in the concentration of taurine in plasma. Major changes in whole blood concentration do not occur as rapidly as plasma and therefore appear to be a superior diagnostic test for taurine status. An inadequate number of samples have been analysed to define a marginal taurine level from whole blood concentration to prevent clinical signs. The minimal dietary concentration of taurine to prevent clinical signs of efficiency is dependent on the type of diet. For commercial expanded (dry) cat foods a concentration of 1200 mg taurine/kg dry matter appears adequate. Higher concentrations are required in canned diets, 2000 to 2500 mg taurine/kg dry matter to supply adequate taurine. The reasons for the higher concentration of taurine required in canned foods is not due to availability of taurine in the classical context. Rather it appears that heating during the canning process produces products which increase enterohepatic loss of taurine.