Cell stimulation

For the stimulation of the RAW264.7 cells, confluent monolayer cells in DMEM containing 10 fetal bovine serum in transportation buffer were stimulated with each reagents for 30 min in 6-well plates. Then the cells were treated with 1 MCi of 3H -taurine for another 30 min. The cells were washed thrice with cold buffer followed by the lysis buffer. The transported radioactive taurine in the lysate was measured by y-counter. For the analysis of nitric oxide, RAW cells were stimulated for 48 h and...

Gene expression in astrocyte primary cultures exposed to an hyperosmotic chemically defined medium

Primary astrocyte cultures were grown from the cerebral cortex of one-day Sprague-Dawley rats until confluence in DMEM medium containing 10 fetal calf serum for three weeks. The cultures were then adapted over three days to a defined medium without fetal calf serum to eliminate exogenous taurine provided by the serum and then submitted to an hyperosmotic medium. In rat cortical astrocyte primary cultures exposed to an hyperosmotic medium, 30 above the isosmotic value, a transient between 1.5 to...

Results And Discussion

Although the experimental model of large and sudden decreases in osmolality had rendered valuable information to elucidate some basic mechanisms of cell volume control, such changes probably never occur in brain under physiological conditions. This is also true during pathological situations such as chronic hyponatremia, water intoxication or the inappropriate secretion of vasopressin, where the osmolarity changes in the brain interstitial space occurs most likely in a gradual manner, as the...

Correlation Between Dietary Taurine Intake And Meat Intake

The relation between dietary meat intake and dietary taurine intake is shown in Figure 5. There was no significant correlation between meat intake and taurine intake. The dataof the subject with highest meat intake is also shown in Figure 5. This female had higher energy intake and protein intake than others. Figure 5. Correlation between dietary taurine intake and meat intake Figure 5. Correlation between dietary taurine intake and meat intake

Introduction

It has been proposed that in non-human, mammalian urinary bladder at least three types of excitatory neurotransmitter elicit detrusor muscle contraction, acetylcholine, ATP and a component which is insensitive to Taurine 4, edited by Della Corte et al. Kluwer Academic Plenum Publishers, New York, 2000. both atropine and suramin. It has been hypothesised that, in small mammals, purinergic transmission is responsible for initiation of voiding, whereas cholinergic transmission affects the...

Discussion

Ication of NMDA caused taurine release as has been shown NMDA receptor antagonists prevented taurine release34 suggesting a mechanism specifically related to activation of that receptor. Co-application of NMDA and the NO synthase inhibitor L-NAME also prevented the release of taurine ointing towards a role of the NO cascade in the taurine release process34. Thus the NMDA-induced taurine release is (at least to a large extent) mediated via the NO cascade. Consequently, the results as obtained in...

Mitochondrial swelling swelling induction and measurement

Following preparation in KSH, the mitochondria (0.5 mg rnl) were suspended in a spectrophotometer cuvette, 1 cm path-length at 25 C, in a medium (195 mM mannitol, 65 mM sucrose, 3 mM HEPES, pH 7.2) containing 10 mM succinate (K+ salt) plus 13 M rotenone. After the absobance at 540 nm stabilised, swelling was induced by Ca2+ (50, 25, or 10 M) plus potassium phosphate (Pi) (5 mM)7. After addition of Pi, the mitochondrial swelling was measured according the method of Broekemeir4, by monitoring the...

HPLC Analysis of DNA Adducts

The mobile phase consisted of (final concentrations) 50 mM sodium acetate, 1mM EDTA and 2 methanol, pH 5.520. The mobile phase was vacuum filtered using nylon filters (0.45 m) from MSI (Westboro, MA) and degassed for 20 minutes under helium. The HPLC apparatus consisted of a PM-11 pump, LC-4B amperometric detector, LC22A temperature controller (Bioanalytical Systems, West Lafayette, IN), and Rheodyne model 7125 injector with a 50 l fixed loop. The column used was a Microsorb-MV, C18, 5 m, 4.6mm...

Methods

PC12 cells were cultured in RPMI media supplemented with 10 horse serum, 5 fetal bovine serum, and penicillin streptomycin. PC12 cells have been used extensively as a model system to test for neuroprotection22. PC12 cells were obtained from ATCC and cultures were used between passage 12 through 27. The cells were cultured until confluent and then plated 24 hours before use in experiments into 96 well plates treated with rat tail collagen. Dilutions based on cell counts using a hemocytometer...

RBC Suspension

Rats were decapitated just before an experiment, and their blood samples collected into heparinized polyethylene test tubes immersed in ice. RBC suspensions were prepared according to the method of Sharma and Premachandra43. After centrifugation of the blood sample at 700 x g for 10 min, and removal of the plasma and buffy coat by aspiration, the pellet was suspended in ice-cold isotonic phosphate-buffered saline (PBSG, 10 mM phosphate buffer pH 7.4-1 50 mM NaCl) solution, supplemented with 5...

Preparation of tissue for electron microscopy

Sections were treated with 1 osmium tetroxide, dehydrated and infiltrated with Ducupan resin that was polymerized at 60 C. Cells and areas of interest were identified and photographed. Sections of 60 nm thickness were taken using the ultra-microtome (Reichert-Jung, Ultracut-E) and collected on Pioloform-coated grids, treated with lead citrate and then examined under the electron microscope. Figure 1. Appearance ofTH- immunoreactive MPP+_-resistant neurons in the striatum The photomicrographs A...

Preface

Expanding the circle A quarter century of research This volume is the selected, edited proceedings of the International Taurine Symposium held at the Certosa di Pontignano, near Siena, Italy in August 1999. The meeting was a satellite to the 17th Biennial Meeting of the International Society ofNeurochemistry which was held in Berlin, Germany, immediately afterwards. The subtitle of this meeting was taken from the fact that the first International Symposium was held in Tucson, Arizona, USA in...

Materials And Methods

Six groups, each of 6 rats were used. Details of the treatments and diets are given Fig. 1 and Table 1 of the accompanying paper8. After 7 weeks the pancreas of each rat was removed and tissue samples were fixed in Bouin solution for paraffin sections, and prefixed in 2.5 glutaraldehyde and postfixed in 1 osmium tetroxide. Following dehydration, the samples were treated with Epon 812. Thin sections were stained with uranyl acetate and lead citrate and examined with a Jeol-100CX electron...

Results Animal Study

The body weight at the end of the feeding period for rats fed the LP diet was significantly lower (p < 0.05) than that for the rats fed the other four diets (Table 4). The LP or basal diet contained (wt wt) 10 casein and -0.3 sulfur amino acids, which provides just below the National Research Council (NRC, 1978) recommended levels of protein and sulfur amino acid intakes for growing rats of 12 protein and 0.6 sulfur amino acids. De- CDO Activity (nmol (min.mg CDO 1*rotdn relative unils'mg CDO...

Methods Animals

Female common cats were fed a completely defined taurine-free synthetic diet (BioServe, Frenchtown, NJ) for at least 6 months prior to mating. The cats were severely taurine-depleted, with plasma taurine concentrations of less than 1 mole 100 ml (0.1 M) Other females (controls), fed the same diet supplemented with 0.05 taurine, maintained plasma taurine levels of approximately 25-50 M. Male cats were fed the taurine-supplemented diet except during mating with females receiving the taurine-free...

Inhibition of taurine transporter activity by rapamycin

RAW264.7 cell line was exposed to immunosuppressive agents, such as rapamycin and CsA, for 5 min and activity of the TAUT was then measured the. When we used 1, 10, 100 and 1,000 ng ml rapamycin in this system, a concentration of 1 ng ml reduced the TAUT activity to 11.9 , when compared to that of the control (Fig. 1). The blocking of the transporter could be induced within 5 min when the cell was cultured with rapamycin for 5, 30, 60 and 300 min. Figl. Effects of rapamycin on taurine...

Conclusion

The regulation of taurine biosynthesis can be summarized as following (i) When neurons are stimulated, the arrival of action potential will open the voltage-dependent Ca2+-channel, resulting in an increase of intracellular free Ca2+, Ca2+ i, (ii) Elevation of Ca2+ i will trigger release of taurine as well as activation of PKC, which in turn activates CSAD through protein phosphorylation (iii) The activated CSAD then synthesizes more taurine to replenish that lost due to stimulation-mediated...

Deletion Analysis of the Proximal 5Flanking Region of the Rat TauT Gene

To determine the basal promoter sequence of TauT, luciferase reporter plasmids were constructed that contained a nested set of deletions extending from nucleotide +48 within the first exon to-124, -269, -574, -963 and -1532 within the proximal 5'-flanking region. Plasmids were transfected into LLC-PK1 and MDCK cells, and luciferase activity was measured. As shown in Fig. 3, luciferase activity was high and quite similar in cells transfected with the constructs having deletions from -1532 to...

Local application of high potassium

Local application of high potassium allways induced a single spreading depression. In parallel, glutamate and taurine increased. The recovery of glutamate was very fast whereas the restoration of normal taurine levels required 30 to 40 min. Application of lubeluzole had no effect. Figure 2. Summary of the effects of Lubeluzole, LNAME, Ketamine or APV on the extracellular levels of taurine released by the application of either K' (128 mM, 3 min, open bars) or NMDA (10 mM, 30 s, black bars)....

Experimental design

Seven week-old mice were grouped into 4 (n 9 per group) Group 1 (control), in which the mice consumed a regular chow Group 2 (control plus taurine), in which the mice consumed a regular chow and 1 taurine Group 3 (high fat), in which the mice consumed a high-fat diet and Group 4 (highhat diet plus taurine), in which the mice consumed a high-fat diet and 1 taurine. At the end of experiment, blood samples were collected from femoral arteries and hearts and aortas were excised. The livers were...

Materialsand Methods Stereotaxic administration of MPP

Female Wistar rats (180 g) were anaesthetised with 2.7 ml kg of a mixture of Hypnoval and Hypnorm in distilled water (1 1 2). Animals received injections of MPP+ at doses of 0.5 nmol, 5.0 mol, 30 nmol and 90 nmol to the right striatum, and sham injections of 0.05 M PBS (phosphate-buffered saline) in the left striatum at the co-ordinates AP 9, L 2.6 and DV 5.2 with reference to Bregma, taken from the stereotaxic atlas of Paxinos and Watson3. The nose-bar was set at -3.3 mm. Doses were given in a...

Studies of chronic alcohol abusers during detoxification

Blood specimens were collected from alcohol abusers n 32 admitted to the National Alcohol Inpatient Unit at the Royal Bethlum Hospital, Beckenham Kent, UK. Ethical permission for these studies was given by the Bethlum & Maudesley NHS Health Trust Ethical Committee. All patients fulfilled ICD-10 DSM IV criteria for alcohol dependence. A blood specimen was taken at the time of the planned admission for the assay of alcohol levels, biochemical parameters as well as haematological indices....

Measurement of serum and liver cholesterol

Serum was prepared by centrifugation at 3,000 rpm for 10 min. Serum cholesterol was measured using an enzymatic method and commercial kits, Autosera CHO-2. HDL cholesterol was measured in the same manner after precipitating out the LDL and VLDL cholesterol with sulfate-Mg2+ 36. LDL plus VLDL cholesterol was calculated by subtracting HDL cholesterol from the total cholesterol. The cholesterol content of the liver was determined enzymaticallyafter extractionwith isopropanol20.

Expression of taurine transporter on raw cells

Murine macrophage cell line, RAW264.7, was used for the measurement of TAUT activity. Confluent cells on 6-well plate was pretreated with unlabelled a- or P-amino acids at a concentration of 1 mM for 10 min and followed by the addition of 1 Ci 3H -taurine for another 30 min. Pretreatment of the cell with unlabelled taurine for 30 min led to blocking of radioactive taurine transport, to 4.8 of the control. Pretreatment with P-alanine also blocked taurine transport, to 12.9 of the control....

Cell volume

No changes in the cell volume were observed in brain slices ofHE rats or slices of control rats treated with ammonia Treatment of slices from HE rats with ammonia led to a significant above control increase ofcell volume. Fig. 3. Effect of HE induced with thioacetamide and or treatment with 5 mM ammonium acetate (ammonia) on the fractional amino acid efflux from cerebral cortical slices. Results are mean SD for 5-6 experiments. *p< 0.05 vs control (one-way ANOVA followed by Dunnet's test)....

Polysome Profile

Figure 4 shows the distribution of CDO mRNA along the liver polysome profile. The distribution of CDO mRNA with polysomes of various sizes was similar in liver of rats fed the low or high protein diets. For cyclophilin mRNA, there was a clear increase in association of the mRNA with poly-somes of higher ribosome number, which paralleled the distribution observed for CDO. Ferritin mRNA, whose translation is regulated in response to cellular iron status, showed markedly less association with...

Amino acid analysis of cardiac tissue

Table 2 shows the results of amino acid analysis of rat cardiac ventricle before and after exposure to hyperosmolality. It can readily be seen that in hypernatraemic animals there is a significant increase in several amino acids, but that taurine is not only the most abundant amino acid identified, but also shows the largest increase. Table 2. The amino acid content mmol kg-1 dry weight of cardiac muscle of rats made chronically hyperosmolal. Modified from Bedford and Leader_ Table 2. The amino...

Initial amino acid content in the slices

HE increased the content of Asp (by 36 ), Glu (by 77 ), Gln (by 29 ) and Tau (by 67 ). Treatment with 5 mM ammonia in vitro increased the content of Glu, Gln and Tau in a degree comparable to the increase evoked by HE, but did not affect the Asp content. Ammonia treatment did not significantly potentiate the effects of HE on the amino acid content. Fig. 2. Effect of HE induced with thioacetamide and or treatment with 5 mM ammonium acetate (''ammonia'') on the amino acid content in the cerebral...

References

Lombardini, J.B., 1991, Taurine retinal function. Brain Res. Rev. 16 151-169. 2. Baylor, D., 1996, How photons start vision. Proc. Natl. Acad. Sci. USA 93 560-565. 3. Finn, J.T., Grunwald, M.E., Yau, K.-W., 1996, Cyclic nucleotide-gated ion channels an extended family with diverse functions. Annu. Rev. Physiol. 58 395-426. 4. Salceda, R., Lopez-Colome, A.M., Pasantes-Morales, H., 1977, Light-stimulated release of 35S taurine from frog retinal rod outer segments. Brain Res. 135 186-191. 5....

Coadministration of ethanol and MK801

When the NMDA inhibitor MK801 alone was administered i.p. to rats, 10mgkg, there was a gradual decrease in hippocampal taurine content which reached significance at 100 minutes and was sustained for the duration of the experiment, Fig. 3. However, when MK80 1 was administered 20 minutes before ethanol injection, 2 g kg, no alteration in the taurine release into the hippocampal microdialysate was assayed during the initial stages, taurine only declining significantly after 200 minutes, Figure 4....

Electronparamagnetic Epr Spectroscopy

EPR spectroscopy allows the assignment of structures to paramagnetic compounds and therefore copper(II) complexes (d9) which have a single unpaired electron give readily resolved spectra. Spectra of the copper(II) -Gly-Gly (1 1) system in the absence and presence of taurine (5-fold excess) at pH values of 5.0, 7.0 and 9.0 were recorded at liquid nitrogen temperature. The g and A parameters, obtained by computer simulation12, are shown in Table 3. As reported previously the copper(II) - Gly-Gly...

Correlation Between Dietary Taurine Intake And Fish And Shellfish Intake

Fish and shellfish are rich in taurine. Therefore, the relation between fish and shellfish intake and taurine intake is shown in this correlation diagram of Figure 4. There was a significant correlation between fish and shellfish intake and taurine intake. Person of highest fish and shellfish intake Taurine intake 1055 mg day fish and shellfish intake 465 g meat intake 0 g fish and shellfish intake 205 g Figure 4. Correlation between dietary taurine intake and fish and shellfish intake Below...

Taurine Fluxes In Insulin Dependent Diabetes Mellitus And Rehydration In Streptozotocin Treated Rats

Rose1, Manju Bushi2, Inderjeet Nagra2and W. Ewart Davies2 'Department of Paediatrics, Heartlands Hospital, Birmingham, England 2Department of Pharmacology, Medical School, University of Birmingham, Birmingham, Abstract The effect of streptozotocin induced diabetes mellitus and rehydration on brain taurine and brain water content was studied in 4 groups of rats. Two groups of rats with diabetes mellitus were used. In one group, taurine and brain water content were determined following...

Animal Feeding Study

Daily weight gain and final body weight of rats fed both the B and B+AA diets were significantly lower (p < 0.05) than that of rats fed the sulfur amino acid-supplemented diets (Table 5). These lower values were anticipated due to the protein sulfur amino acid content of the diets as reviewed above. The B diet did support a reasonable rate of weight gain, and food intake was similar (p > 0.05) to that of rats fed the sulfur amino acid-supplemented diets. The daily intake of rats fed the...

Local application of NMDA

In a first set of experiments, two short pulses of NMDA (10 mM, for 30 s, via perfusate) were given with an inter-stimulus interval (ISI) of 60 min (Fig. 1). After the first' and the second NMDA bolus, a single spreading depression always could be observed. Blood pressure, heart rate and blood gases did not change after application of NMDA. Figure 1. Representative experiment showing the effect of local NMDA application (1 0 mm, 30 s, indicated by arrow) on DC (line graph) and dialysate...

Rat retinal cells

Protein kinase C activator or phosphatase inhibitor effects. Adult retinal cells incubated in the presence of 1 M PDBu or 10 nM OKA for 10 to 120 min did not present any significant difference in 3H taurine transport with respect to corresponding control. The treatment of these cells, cultured for 24 h, for 1 h with different concentrations of PDBu did not produce significant modifications. However, OKA produced a dose-dependent decreased in the uptake of the amino acid into rat retinal cells...

Phmetric Studies

The complex Cu(Gly-GlyH_1) may react with taurine to give a ternary species in which taurine acts as a monodentate ligand (Fig 4) or in which the sulphonate group is also axially coordinated and taurine acts as a bidentate ligand. As was previously confirmed, in a study of the interactions of Cu(Gly-GlyH-1) and Cu(Gly-AspH-1) - with B-alanine, this axial coordination cannot occur in the latter case. The significantly larger stability constant for the formation of the ternary complex...

Altered Muscle Protein Function In The Taurinedepleted Heart

Very little information is available on the status of the muscle proteins in the taurine depleted heart. In one study, Lake25 reported that the GES Figure 3. Calcium transient profile of osmotically stressed cardiomyocytes. Cardiomyocytes were incubated for 3 days in serum-substituted medium containing either 5 mM glucose (Control Cell), 30 mM glucose (High Glucose Cell), 30 mM mannitol (Mannitol Cell) or 5 mM B-alanine (B-alanine Cell). The cells were loaded with 2.0 M Indo-1, a fluorescent...

Quantitative assessment of atherosclerotic lesions

Quantification of atherosclerotic lesions was carried out as previously described16'17. After exsanguination of ether anesthesed mice, the heart was perfused through the left ventricle, first with phosphate-buffered saline and then with 10 buffered formalin, the right atrium were cut to allow the perfusate to flow out. The heart and aorta were immersed in 10 buffered formalin. The formalin-fixed aorta linked to the heart was opened, the adventitia separated and all three aortic valves were...

Gene expression in the renal papilla of rats undergoing diuresis and antidiuresis

The osmolarity of excreted urine was experimentally modified by submitting the rats to diuresis followed by antidiuresis. The rats were given water containing 10 sucrose for 10 days. This treatment brought about chronic diuresis. The animals excreted large volumes of diluted urine. Half of the rats were maintained under a similar diuresis while the other half were submitted to water deprivation for 2 days. This treatment brought about antidiuresis. The animals excreted small volumes of...

New Hplc Evidence On Endogenous Tauret In Retina And Pigment Epithelium

Haroutounian, Thomas E. Gundersen1, Rune Blomhoff, Kjell Fugelli2 & Hilde Kanli2 Buniatian Inst. ofBiochemistryofNatl. Acad. Sci. of Armenia, Sevag St. 5 1, Yerevan 14, Armenia 1Inst. for Nutrition Res., University of Oslo P.O.Box 1046, 0316 Oslo, Norway 2 Div. of Gen. Physiology, University of Oslo P.O. Box 1051 Blindern, 0316 Oslo 3, Norway Abstract This investigation was improve the separation for tauret (retinylidene taurine) and to compare its content...

Surgery and HPLC methodology

Male Wistar rats, 200-250g, 6 in each group, were anaesthetised with chloral hydrate (400mg kg). A guide cannula was inserted into the hippocampus CA2 region using standard stereotaxic techniques (A P -4.3 mm M L 4.0 mm D V -3.0 mm) of Paxinos and Watson11. The dialysis experiments commenced 48h post operation. The probe was connected to a micro-infusion pump (Infusion syringe pump 22, Harvard apparatus) and continuously perfused at l min with Ringer solution containing 145 mM NaCl 4 mM KCl 1.3...

Regulation Of Taurine Biosynthesis

Contrary to taurine transporters, which have been cloned and characterized133661, little is known regarding the molecular mechanisms in the regulation of CSAD activity. Jerkins and Steele26 reported that the hepatic CSAD activity was decreased in female rats, adrenalectomized rats and rats fed L-methionine. The decrease of CSAD activity was due to reduction in CSAD protein. The same authors also reported that CSAD activity in liver of rats fed with sulfur amino acids was markedly reduced24,...

Release Of Taurine From Son Astrocytes

Application of hypotonic stimuli increases the basal release of both endogenous taurine12 and pre-loaded 3H -taurine from acutely isolated supraoptic nuclei from adult male rats (Fig 1A)7. Both uptake and release of 3H -taurine are inhibited by the selective glial metabolic blocker fluorocitrate, whereas they are unaffected by the elimination of all SON neurons by an excitotoxic treatment with NMDA7, in agreement with the specific astrocytic localization of endogenous taurine in this...

Results

Fig. 2 shows the effects of taurine on mitochondrial swelling induced by 5 mM Pi plus Ca2+ at different (50, 25 and 10 M ) concentrations in mitochondria energised by 10 mM succinate. The addition of Pi in the presence of Ca2+ was followed by a fast and extensive Ca2+-dependent decrease of absorbance, indicative of mitochondrial swelling. The absorbance change correlated with the concentration of Ca2+ and was completely reversed by 0.5 M cyclosporin A (CsA), but was not affected by 10 mM...

Results In Vitro Experiments

The experimental conditions used to carry out the various in vitro studies with PHZ were established on the basis of the release of K+ from RBCs as a function of the incubation time, concentration of added PHZ and the timing of the pretreatment with the doses of TAU listed in Figs. 1 to 3. From these studies, an incubation time of 60 min, a concentration of TAU (or a related sulfur compound) of 150 mM, and a concentration of PHZ of 75 mM in the RBC suspension were adopted. Because of assay...

Results Tissue Ultrastructure

After treatment normoxic rats survived well, hyperoxic rats suffered and one of the saline-treated group died. Hyperoxic treatment caused tissue damage in the untreated group, with diffused hematoma in liver and lungs and edema in the thorax. Hypotaurinetreated hyperoxic rats and, to a lesser extent, taurine-treated ones appeared normal from the anatomic point of view. It is well known that hyperoxic treatment causes severe damage to mitochondria and membrane derangement. In our study,...

Resultsand Discussion

The hydrolytic activity of MMP-9 activated by various amounts of LPS (lng ml-10000ng ml) in murine PEC is shown in Fig 1. EDTA inhibited the hydrolytic activity of MMP-9 by chelating Zn ++ and Ca ++ that is required for the reaction. 1,10-phenanthroline, which chelates divalent metals, also inhibited MMP-9 activity. However, PMSF, an inhibitor of serine protease such as trypsin and chymotrypsin, did not inhibit MMP-9 activity, consistent with the characteristics of this enzyme620. There was no...

Role of Osmotic Stress in Altered Calcium Transport

Another action of taurine that may contribute to altered calcium transport in the taurine depleted heart involves an osmotic linked mechanism. Car-diomyocytes exposed to medium containing 5 mM B-alanine lose 50 of their intracellular taurine pool, a large enough change to affect the osmotic balance across the cell membrane and trigger a volume regulatory mecha-nism44. This compensatory mechanism is important to the cell, not only because it restores cell volume and osmotic balance, but because...

Sarcoplasmic Reticulum Isolation and Calcium Uptake Measurements

Both EDL muscles of male Wistar rats aged 3 months were quickly removed and homogenized at 0 C in 1 ml of a buffer solution (200 mM sucrose, 10 mM sodium azide, 1 mM EDTA, 40 mM L-histidine, pH 7.8). The light fraction of sarcoplasmic reticulum (SR) was isolated according to the method of O'Brien13. The rate of Ca2+ uptake by the SR was determined with the Ca2+ fluorescent indicator fura-2510. The assay was initiated by injection of 15 g SR in 200 l assay medium which contained 10 M Ca2+, 5 M...

Serum and tissue analysis in chronic hyperosmolality

Serum osmotic pressure and ion concentrations of cardiac tissue of rats subjected to chronic hyperosmolality are shown in Table 1. Cellular shrinkage by elevation of serum NaCl or glucose over a 4-5 day period is accompanied by an accumulation of cellular osmoles and, in consequence, a partial mitigation of cellular shrinkage. In contrast, elevation of serum osmolality by heightened urea levels following bilateral nephrectomy does not result in either cellular shrinkage nor the accumulation of...

Skinned Fiber Preparation and Ca2 Sensitivity Determination

Bundles of rabbit psoas muscle were chemically skinned as described by Brenner2. Bundles of about 10 fibers were dissected and mounted on a force transducer. Sarcomere length was measured by laser diffraction1 and adjusted to 2.2 m. Fibers were stimulated by immersion in a solution containing 0.54 M buffered free Ca2+, 5 mM EGTA CaEGTA, 10 mM imidazole, 5 mM MgATP, 10 mM caffeine, 20 mM taurine or 20 mM GES, pH 7.2 and ionic strength x 2 adjusted at 138 mM. Fiber bundles were relaxed between...

Slice Preparation

Experiments were performed on 400-pm-thick transverse hippocampal slices obtained from adult female Sprague-Dawley rats (200-250 g), following standard methods. Briefly, after the rat was decapitated, its brain was rapidly removed and dropped into ice-cold standard medium (composition in mM NaCl 119, NaHCO3 26.2, KCl 2.5, KH2PO4 1, MgSO4 1.3, CaCl2 2.5 and glucose 11) pregassed with 95 O2 and 5 CO2. Once the hippocampi were dissected out, they were sliced by a manual tissue chopper and placed...

Spectrophotometry Analysis of Quinones

In order to address the mechanism of action of taurine and related analogs in the reduction of oxidative damage to DNA, an experiment was performed to measure quinone formation from the oxidation of L-dopa. Taurine and taurine analogs were all tested at a concentration of 10 mM. Reaction conditions were identical to those described for the DNA damage assays, except at the end of the 15 minute incubation period the samples were immediately read in a spectrophotometer at 490 nm to measure...

Statistical Analysis

All samples were run in duplicate and each value of N represents an independent replication. An N 5-6 was used in all experiments. In pilot studies, mean concentrations of the adducts (nmol adduct nmol guanine per sample) were calculated. The mean adduct concentrations from the damaged samples were compared to the mean adduct concentrations from the damaged with taurine. Statistical comparisons were made to determine the protective effects of taurine in reference to individual adduct...

Statistical Analysis of the Data

The experimental results were analyzed for statistical significance by unpaired Student's t-test, one-way analysis of variance (ANOVA), and Tukey's post-hoc test using a commercial software program (StatMost for Windows 2.01 from Datamost Corporation, Salt Lake City, UT). Differ-ences were considered as statistically significant at p 0.05. Values were reported as the means SEM for n 6. Table 1. Effect of sulk-containing compounds on PHZ-induced K+ leakage, MDA formation, GSH depletion and...

Subjects

Twenty one male patients, aged 75 to 95 years, sufferhg from coronary heart disease, multiple cerebral infarction, cancer, subdural hematoma or respiratory failure, were grouped according to the duration of tube feeding, as shown in Table 1. Table 1. Characteristics of old aged male patients undergoing long term enteral nutrition (EN) Table 1. Characteristics of old aged male patients undergoing long term enteral nutrition (EN) * Figures represent mean value SD ranges in brackets. * Figures...

Surgical procedure

Wistar male rats (220-280 g) were anaesthetised with urethane (1.80 g kg) and fixed in a stereotaxic frame. A burr hole of 3 mm diameter was drilled through the skull. A microdialysis (MD) probe (2 mm length, 0.5 mm outer diameter, Carnegie Medicin, Stockholm Sweden) wasimplanted in the parietal cortex. As perfusate artificial cerebrospinal fluid (aCSF composition NaCl 125 mM KCl 3 mM CaCh 1.1 mM MgCh, 0.8 mM N2HPO4, 0.5 mM NaHCO3 25 mM D-glucose 6 mM) was used at a flow rate of 2 l min....

Swellinginduced Leak Pathways

If mammalian cells are exposed to a hypotonic solution, they initially swell as almost perfect osmometers. They reach a maximal degree of cell swelling within the first minutes, and thereafter they regulate their volume back towards the initial value. This back regulation, termed regulatory volume decrease (RVD), involves net loss of KCl and net loss of organic osmo-lytes. Taurine is an important organic osmolyte in mammalian cells, and loss of taurine is often taken to indicate a reduction in...

Synthesis of chlorotaurine

Chlorotaurine was synthesized freshly each day by reaction of taurine and NaOCl in a 1 1 molar ratio. Chlorotaurine formation and hypochlorite disappearance were monitored by W absorbance, absorbance maxima at 252 nm ( 429 M-1.cm-1), and 291 nm ( 142 M-1. cm-1), respectively9. The first order decay of chlorotaurine to sulphoacetaldehyde could then be measured by HPLC after incubation of various concentrations of chlorotaurine (0 to 3.65 mM) in 0.1 M phosphate buffer for various times at 37 C,...

Synthesis Of Taurine Analogues From Alkenes

Preparation of 2-aminocycloalkanesulfonic acids Fabrizio Machetti, Martina Cacciarini, Fernando Catrambone, Franca M. Cordero, *Maria Frosini, Francesco De Sarlo Dipartimento di Chimica Organica U. Schff , and Centro di Studio sulla Chimica e la Struttura dei Composti Eterociclici e loro Applicazioni, C.N.R., Universit di Firenze, Via G. Capponi 9, 1-50121 Firenze, Italy. * Istituto di Scienze Farmacologiche, Universit di Siena, Italy Abstract ( )trans 2-Aminocyclohexanesulfonic acid and (...

Taurine and skeletal muscle potassium channels

K+ channels comprise a large family of genetically distinct entities with distinct biophysical properties and physiological roles. In heart taurine modulates the function of many types of K+ channels, such as delayed rectifier K+ channels, important for action potential duration, and ATP sensitive K+ channels (Katp) that in excitable tissues couple cell metabolism to membrane electrical activity2223. Katp channels are abundantly present on sarcolemma of striated muscle fibers, where they exert...

Taurine and skeletal muscle sodium channels

It is well known that voltage-gated Na+ channels play a pivotal role in excitable tissues for the generation and propagation of the action potential. Genetic defects of skeletal muscle Na+ channels result in changes of excitation pattern and muscle contractility being responsible for recessive hereditary disorders ranging from myotonias to paralyses. Figure 2. Effect of in vitro application of taurine on resting macroscopic chloride conductance of fast-twitch extensor digitorurn longus (EDL)...

Taurine Composition Tables

In preparing taurine composition tables of animal food and algae, data on 91 foods containing taurine were collected from reference papers. Also, 51 processed foods and others were analyzed for taurine content (mg l00g). In this analysis, food samples were first homogenized with 3 sulfosalicylic acid solution, then each homogenate was centrifuged to obtain the supernatant for an amino acid analysis. An automatic amino acid analyser determined the concentrations of the amino acid.

Taurine Intake And Excretion In Patients Undergoing Long Term Enteral Nutrition

Chen2 'Dept. Food Science andNutrition, Dankook University, Seoul 140-71 4, Korea, 2Institute of Sports Medicine, The Third Teaching School of Clinical Medicine, Beijing Medical University, Beijing 100083, China Abstract The purpose of this study was to investigate whether serum concentration and urinary excretion of taurine are influenced by marginal taurine intake. Twenty one male patients (75 to 95 years old), suffering from coronary heart disease, multiple...

Taurine uptake assay

The assay used in measuring taurine uptake was performed as previously described13. Briefly, the reaction was performed in KBR solution and for uptake kinetic experiments, unlabelled taurine was added in varying concentrations (10 M - 10 mM). For other experiments, unlabelled taurine was added to a final concentration of either 50 M or 1.5 mM. Equal amounts of 3H taurine were added to each test tube (1 Ci) and incubated in a 37 C water bath for 2 minutes. The reaction was initiated by the...

TG Polymorphism is a Strong Enhancer for the Promoter Function of the Rat Taurine Transporter Gene

To determine the role of the TG repeat (TG22) that is found upstream of the basal promoter DNA sequence (from -19 1 to -235), a p-177 reporter with TG repeat deletion was prepared by PCR. As shown in Fig. 5, transient transfection of the p-269 reporter (which contains the TG repeat) into MDCK and LLC-PK cells showed a 100- to 200-fold increase in luciferase reporter gene expression, as compared with control (pGL3-Basic vector). However, transient transfection of the p- 177 reporter, which does...

The Biological Sulfur Cycle

The biological sulfur cycle, showing the various oxidation states for reduction and oxidation (odd-numbered oxidation states require dimeric intermediates). Sample oxidation states are -2 sulfide 0 sulfur +2 sulfinate, thiosulfide +4 sulfonate, taurine +6 sulfate. Major entry points of inorganic sulfur into the cycle are indicated in gray boxes. Mammals are capable only of sulfur oxidation. Plants, bacteria and fungi can perform the complete cycle. Unlike the rest of the biosphere,...

The Mitochondrial Permeability Transition And Taurine

'Mitri Palmi, 'Gisele Tchuisseu Youmbi, 'Giampietro Sgaragli, 'Antonella Meini, 'Alberto Benocci, 'Fabio Fusi, 'Maria Frosini, 'Laura Della Corte, 3Gavin Davey and 3Keith Francis Tipton 'Istituto di Scienze Farmacologiche, Universit di Siena, Siena, Italy 2Dipartimento di Farmacolog a Preclinica e Cl nica, Universit di Firenze, Firenze, Italy 3Department of Biochemistry, Trinity College, Dublin, Ireland Abstract Perturbed cellular calcium homeostasis has been implicated in both apoptosis and...

The Possible Role Of Taurine And Gaba As Endogenous Cryogens In The Rabbit

'Maria Frosini, 'Casilde Sesti, 'Mitri Palmi, 'Massimo Valoti, 'Fabio Fusi, 'Piero Mantovani, 2Loria Bianchi, 2Laura Delia Corte and 'Giampietro Sgaragli 'Istituto di Scienze Farmacologiche, Via Piccolomini 170, Siena, Italy 2Dipartimento di Farmacolog a, Viale Pieraccini 9, Florence, Italy Abstract We investigated whether heat-stress induced hyperthermia could enhance release of both endogenous taurine and GABA from nerve cells into the extracellular compartment, thus acting like endogenous...

The relationship between intracellular taurine content and taurine cardioprotection against ischemic injury

The myocardial taurine content is elevated in response to the chronic stresses that trigger the development of heart failure1819. Conversely, in acute episodes of stress, myocardial taurine levels decline2021. There is no evidence that elevated levels of taurine are harmful to humans, As shown in Figure 5, exposure of the cells to 20 mhM taurine for 24 h elevated the intracellular taurine content from 240 pmol to 1600 pmol mg protein. (A) HPLC Analysis Profile of Taurine (B) Taurine...

Time min

Effect oftaurine on mitochondrial membrane depolarisation induced by Ca2+ and Pi. Mitochondria (lmg ml) were incubated in medium (195 mM mannitol, 65 mM sucrose, 3 mM HEPES, pH 7.2) containing 13.3 M rotenone, in the presence or absence of 10 mM taurine. AY was measured by using a TPMP+-ion selective electrode. After the calibration, 10 mM succinate (K+ salt) was added, followed by addition of50 M Ca2+. At steady-state Ca2+ accumulation, addition of 5 mM Pi was made. All traces are...

Transcription Factor Sp1 is Critical for the Basal Promoter Function of TauT

The TauT basal promoter sequence contains four potential Sp1 sites, located at -162 to -168, -155 to -150, -144 to -139, and -133 to -128 in relationship to the transcription start site. To test if Sp1 transcription factor is Figure 3. Deletion analysis ofthe proximal 5'-flanking region ofthe TauT gene. Recombinant plasmids containing nested deletions of the proximal 5'-flanking region of TauT (in pGL3-Basic) were generated by PCR. A (top) Constructs are numbered according to the position...

Treatment of DNA

DNA was damaged using a Fenton type reaction with FeCk, H2O2 and L-dopa, similar to the method described by Spencer et al.12. Reaction mixtures contained (final concentrations) PBS (phosphate buffered saline) (pH 7.4), calf thymus DNA (100 g tube), H2O2 (2.8mM), FeCl3 (60 M), and L-dopa (100 M). Taurine or taurine analogs were tested for their effects on oxidative damage to DNA. Control tubes contained only DNA and PBS. The final assay volume was 1 ml, and reaction mixtures were aliquoted into...

Uptake Experiments in Isolated and in Cultured Cells

3H Taurine (Amersham, 24.1 Ci mmol -200,000 dpm) was used with non-radioactive taurine in concentrations from 0.003 to 100 M. Time-dependent and inhibition experiments utilizing B-alanine, hypotaurine, or GABA, 0.1 nM to 3 mM, were performed in the presence of 40 nM 3H taurine. The cell preparation (250 300,000 per ml) was preincubated at 25 C for 2 min in Ringer solution. Incubation was started by the addition of the substrate. After 2 min the process was stopped by filtration through glass...

Uptake of 3Htaurine into oocytes

3 days after cRNA injection into Xenopus oocytes, single oocytes were used for determination of taurine uptake. Oocytes were incubated for 4 hours in standard saline containing 3H-taurine. For experiments with drugs and inhibitors, the bathing solution contained taurine at a concentration of 0.1 mmol, for periods up to 4 hours. At the end of each experiment the oocytes were washed rapidly three times in taurine-free solution, external fluid blotted free, and, after lysing in 10 sodium dodecyl...

Visible Spectra

The formation of a ternary copper(II) - Gly-Gly - taurine complex in aqueous solution was confirmed by visible spectroscopy. UV-visible spectra of the copper(II) - Gly-Gly and copper(II) - Gly-Gly - taurine (1 1 1 and 1 1 8) systems were recorded at various pH values. The values and molar extinction coefficients as a function ofpH are summarised in Table 2. The Xmax values for the copper(II) - Gly-Gly system do not show a pH dependence in the pH range studied as the two species present in this...

Western and Northern Blot Methods

The purified IgG fraction from rabbit anti-CDO serum and the cDNA for CDO were gifts from Dr. Yu Hosokawa (National Institute of Health and Nutrition, Tokyo, Japan)12. The cDNA fragment was cloned into pBluescript SK+. A partial CDO cDNA was prepared using complementary DNA probes corresponding to bp 459-717 of rat liver CDO13. DECAprobe template-actin-mouse and DECAprobe tem-plate-18S-mouse (Ambion, Inc., Austin, TX) were used as internal standards for Northern...

Taurine and calcium ion availability in skeletal muscle

As already stated taurine is able to control excitation-contraction (e-c) coupling in cardiomyocytes by modulating Ca2+ channels and therefore the influx of Ca2+ that is pivotal for cardiac contraction22. As a consequence all the other Ca2+-dependent mechanisms are affected26. The e-c coupling of skeletal muscle fiber is almost independent of external Ca2+ nonetheless, taurine modulates intracellular Ca2+ levels. As proposed by Huxtable, intracellular taurine can bind, with a low affinity, to...

Cloning of the 5Flanking Region of the Rat Taurine Transporter Gene

The rat P1 genomic DNA library was screened (Genome Systems, Inc., St. Louis, MO) by polymerase chain reaction (PCR) using three sets of Taurine 4, edited by Della Corte et al. Kluwer Academic Plenum Publishers, New York, 2000. oligonucleotide primers based on the DNA sequence of rat brain taurine transporter cDNA (rB16a)17. The primers used for generating the PCR probes were sense primer 5'-GCCAACGCCGCGATCGCCGCCAA-3 ' antisense primer 5'-CTCCTCGTTTTGCTTGAGAGGC-3 ' sense primer 5'-AT...

Materials And Methods Chemicals

All the amino acids and sulfur-containing compounds used in the study were obtained from local commercial sources TAU, hypotaurine (HYTAU), isethionic acid (ISA), N-acetylcysteine (NACY S), L-cysteic acid (CA) and aminomethanesulfonic acid (AMSA) were from Sigma Chemical Co., St. Louis, MO ethanesulfonic acid sodium salt (ESA) and 1,2-ethanedisulfonic acid (EDSA) were from Aldrich Chemical Co., Milwaukee, WI B-alanine (BALA) and N-(2-acetamido)-2-aminoethanesulfonic acid (ACES) were from TCI...

Effects Of Long Term Treatment With Taurine In Mice Fed A High Fat Diet

Improvement in cholesterol metabolism and vascular lipid accumulation by taurine Shigeru Murakami, Yukiko Kondo, and Takatoshi Nagate Medicinal Research Laboratories, Taisho Pharmaceutical Co.Ltd., Ohmiya, Japan Abstract Hypocholesterolemic effects of taurine in rats fed a high-fat and high-cholesterol diet are well established. However, there are few studies on long-term effects of taurine on cholesterol metabolism. In the present study, taurine was dissolved in drinking water and given to...

Appendix Some Taurine Analogues Referred To In This Book

(a) Some metabolites and natural analogues 2-aminoethane-sulfonic acid NH2-CH2-CH2-SO3H 2-aminoethane-sulfinic acid NH2-CH2-CH2-SO2H (hypotaurine) (CYSTEIC ACID) cysteinesulfinic acid 2-amino-3-hydroxy-1 -propanesulfonic acid HOOC-CH(NH2)-CH2-SO2H N-(1-carboxyethyl)-taurine cerilipin CO-NH-CH2-CH2-SO3H (b) Substitutions and simple modifications

Cell Culture and Taurine Transport

RAW 264.7 cells were cultured and maintained in 90 mm culture dishes in DMEM medium containing 10 heat-inactivated fetal bovine serum (FBS) and penicillin (100 U ml) streptomycin (100 Mg ml), and in 6-well plates for taurine uptake experiments. Cultures were maintained at 37 C in 5 CO2 incubator until they reached confluence. Taurine transportation study was performed according to the previous report21, i.e., when the cells were confluent on the dish, the medium was removed from the monolayer...

Participants

Fernanda Amicarelli Dip. di Biologia di Base Universita dell' Aquila Via Vetoio Dr. Junichi Azuma Dept. Clin. Evalut. Med. & Therapeutic Osaka University 1-6 Yamadaoka, Suita-shi Osaka 565-0871 Dr. Steven I. Baskin USA Medical Research Inst. of Chemistry 3 100 Ricketts Point Road Aberdeen Proving Grd-EA., MD Dr. Jennifer J. Bedford Dept. of Physiology University of Otago P.O. Box 913 Dunedin New Zealand Ms. Francesca Bellucci 1 st. Scienze Farmacologiche Universita di Siena Via ES...

The Functions Of Taurine Bile Salt Conjugation

One clear mammalian function of taurine is a metabolic one that of bile salt synthesis. In addition, taurine can conjugate with various xenobiotics. In mammals, xenobiotic conjugation seems to be a relict pathway, significant for certain compounds in certain species but in general quantitatively minor compared with GSH, sulfate, and glucuronide conjugation. Invertebrates contain surface tension-lowering substances in the alimentary canal. These are typically sulfate or taurine conjugates with...

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Sulfur Metabolism Humans

Mammalian transsulfuration. Figure 7. Cysteine metabolism in mammals. The Functions and Metabolism of Cysteine The functions of cysteine include protein synthesis and GSH synthesis for redox and conjugation functions. Cysteine is both neurotoxic and readily oxidized to cystine, a poorly soluble compound. GSH serves as a transportable form of extracellular cysteine as it is less susceptible to oxidation. Cell cysteine levels are lower than GSH levels. Cysteine acts at the NMDA...

Recovery of taurine transporter activity by ionomycin

AS the intracellular concentration of Ca2 is important for the activation of macrophages, RAW264.7 cells were co-treated with rapamycin and ionomycin IM simultaneously. The concentration of rapamycin was fixed at 1 ng ml and the concentration of IM was increased 10-fold from 20 to 2,000 ng ml. When the concentration of IM was increased in the rapamycin-treated cells, the recovery of the activity of TAUT was not altered by the addition of IM Table 11 . However in the case of co-treatment with...