Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “AMINOISOBUTYRIC ACID”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 649 records · Page 36Linked to original sources
PubMed1994

Differentiation of two classes of "A" system amino acid transporters.

Injection of mRNA from GF-14 cells (a mouse lymphocyte cell line) into Xenopus oocytes led to the expression of a transport activity characteristic of amino acid transport system A, i.e., Na+ dependent and recognizing aminoisobutyric acid and its methyl derivative (AIB and meAIB). Sucrose density gradient fractionation of GF-14 mRNA showed that the maximum level of expression of activity was associated with a 2.2-kb RNA fraction. Pretreatment of GF-14 cells with insulin caused a twofold increase in system A transport activity in the cells themselves and the mRNA from the insulin-treated cells induced a comparable increase in A system transport over control mRNA when expressed in oocytes. mRNA from cells treated with insulin in presence of actinomycin D did not show a response to insulin, suggesting that GF-14 cells synthesized new transporter in response to insulin. Similar experiments with mRNA from the Ehrlich ascites cells, showed that expression of system A type transport was associated chiefly with a 4.2-kb mRNA fraction. Although insulin treatment of Ehrlich cells also caused an increase in A system transport activity in the cells themselves (nearly twofold), the response to insulin was not blocked by actinomycin D or cycloheximide. Moreover, mRNA from insulin-treated and untreated Ehrlich cells gave the same level of expression in oocytes of A type amino acid transport. In contrast to GF-14 cells, Western blots of plasma membranes showed that insulin treatment of Ehrlich cells increased the amount therein of a 120- to 130-kDa peptide, previously associated with amino acid transport (Proc. Natl. Acad. Sci. USA 85, 7877, 1988), suggesting that in the Ehrlich cells, but not in GF-14, cells this polypeptide was translocated from intracellular sites in response to insulin. The data associate two distinctly different responses to insulin with A-type amino acid transporters and are consistent with the existence of more than a single A type amino acid transporter in mouse cells.

Amino Acids↗
PubMed1995

Volume-activated amino acid efflux from term human placental tissue: stimulation of efflux via a pathway sensitive to anion transport inhibitors.

The effect of a hyposmotic challenge and hence cell-swelling upon the efflux of a variety of solutes from isolated human placental tissue has been examined. A hyposmotic shock increased the fractional release of taurine, the most abundant free amino acid in placental tissue, via a pathway sensitive to niflumic acid, DIDS (4,4'-Diisothiocyanatostilbene-2',2'-disulphonic acid,) NPPB (5-Nitro-2(3-phenylpropylamino)benzoic acid) and DIOA (R(+)[2-n-butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1H-inden -5-y) oxy] acetic acid). In contrast, tamoxifen was without effect. The cell-swelling induced efflux of taurine was attenuated (40 per cent) by replacing external Cl- with NO3-. The efflux of glutamic acid was also markedly increased by a hyposmotic challenge. Niflumic acid inhibited both basal and volume-activated glutamic acid efflux. A hyposmotic shock also increased alpha-aminoisobutyric acid efflux but not that of 3-O-methylglucose and SO4(2)-. The results suggest that the human placenta can respond to cell-swelling by releasing organic osmolytes such as amino acids via a pathway which is sensitive to anion transport inhibitors. However, it appears that the volume-activated amino acid transport system is independent from the placental anion-exchange pathways. The efflux of these compounds may act with K+ and Cl- efflux to effect a regulatory volume decrease in placental tissue. In addition, volume-activated transport may play a role in transplacental amino acid transfer.

3-O-Methylglucose↗
PubMed1986

An autocrine/paracrine role for insulin-like growth factors in the regulation of human placental growth.

Tissue derived from preterm (9-19 weeks gestation) and term (38-41 weeks gestation) human placentae were examined for their ability to synthesize and secrete insulin-like growth factors (IGFs) in organ culture. IGF-I was measured by a specific RIA, and IGF-II by a rat placental membrane radioreceptor assay. First, explants of placental tissue were maintained in organ culture. These explants secreted immunoreactive IGF-I (IR-IGF-I). There were no differences in the IR-IGF-I content of media conditioned by term and preterm placentae under these conditions. The similarity of this material to authentic human IGF-I was supported by parallel displacement in a specific RIA and coelution during Sephadex G-50 gel filtration. Second, monolayer cultures of fibroblasts from normal human preterm placentae (15-19 weeks gestation) were established. Confluent monolayers of these fibroblasts secreted IR-IGF-I (3-10 pg/10(5) cells X 40 h). IR-IGF-I secretion was reversibly inhibited by 5.3 microM cycloheximide, suggesting that the IR-IGF-I was the result of de novo protein synthesis. IR-IGF-I secretion was stimulated 5-fold by platelet-derived growth factor (0.6 U/ml). The response of monolayers of placental fibroblasts to IGF-I also was tested. IGF-I stimulated alpha-[3H]aminoisobutyric acid transport in these fibroblasts, with half-maximal stimulation occurring at 2-3 ng/ml. Stimulation of alpha-[3H]aminoisobutyric acid uptake by IGF-I correlated with specific binding of [125I]iodo-IGF-I. Half-maximal inhibition of [125I]iodo-IGF-I binding occurred at 2-3 ng/ml IGF-I. Placental tissue also secreted IGF-II-like activity, as measured by radioreceptor assay. Media conditioned by placental explants contained 15-20 ng/mg protein X 48 h, and media conditioned by placental fibroblasts contained 3-7 ng/10(5) cells X 40 h IGF-II determined by radioreceptor assay. These data support the hypothesis that the human placenta produces IGFs (IGF-II and/or IGF-I) that act locally to regulate placental growth.

Aminoisobutyric Acids↗
PubMed1991

Blood-brain barrier disruption after cardiopulmonary resuscitation in immature swine.

We investigated blood-brain barrier permeability in 2-3-week-old anesthetized pigs during and after cardiopulmonary resuscitation. We assessed permeability by tissue uptake of radiolabeled aminoisobutyric acid, after correcting for plasma counts in tissue with radiolabeled inulin. Among 14 regions examined, the transfer coefficient of aminoisobutyric acid in nonischemic control animals ranged from 0.0018 +/- 0.0001 ml/g/min in diencephalon to 0.0049 +/- 0.0003 ml/g/min in cervical spinal cord. After 8 minutes of cardiac arrest followed by either 10 or 40 minutes of continuous sternal compression, there was no increase in the transfer coefficient. Likewise, during the immediate period after ventricular defibrillation, there was no increase in transfer coefficient despite the brief, transient hypertension. However, after 8 minutes of arrest, 6 minutes of cardiopulmonary resuscitation, and 4 hours of spontaneous circulation, the transfer coefficient was significantly increased by 59-107% in 10 of 11 regions rostral to the pons. Plasma volume in tissue measured by inulin was not elevated, suggesting that the increased transfer coefficient was not due to increased surface area. Thus, after an 8-minute period of complete ischemia, the blood-brain barrier remains intact during and immediately after resuscitation despite large vascular pressure fluctuations. However, in contrast to previous work on adult dogs, immature pigs are prone to a delayed increase in permeability, thereby allowing circulating substances greater access to the brain.

Aminoisobutyric Acids↗
PubMed1992

Blood-brain barrier permeability changes after experimental subarachnoid hemorrhage.

Basic mechanisms underlying cerebrovascular permeability responses to subarachnoid hemorrhage (SAH) are still to be defined in detail. Previous investigations examining the occurrence of blood-brain barrier (BBB) breakdown after SAH in the experimental setting have yielded conflicting results. In a rat model of SAH, we assessed BBB changes by means of the quantitative [14C]-alpha-aminoisobutyric acid technique. Experiments were carried out on the second day post-SAH. In blood-injected rats [14C]-alpha-aminoisobutyric acid transport across the BBB increased significantly in cerebral cortices and cerebellar gray matter, averaging 1.3 to 1.5 times control values. The present data indicate that SAH induces well-defined changes in BBB function, possibly involved in the pathogenesis of post-SAH cerebral dysfunction in humans. Results reported here have also potential clinical implications for the management of aneurysm patients.

Aminoisobutyric Acids↗
PubMed2001

Crystal-state conformation of Calpha,alpha-dialkylated peptides containing chiral beta-homo-residues.

Secondary structure formation and stability are essential features in the knowledge of complex folding topology of biomolecules. To better understand the relationships between preferred conformations and functional properties of beta-homo-amino acids, the synthesis and conformational characterization by X-ray diffraction analysis of peptides containing conformationally constrained Calpha,alpha-dialkylated amino acid residues, such as alpha-aminoisobutyric acid or 1-aminocyclohexane-1-carboxylic acid and a single beta-homoamino acid, differently displaced along the peptide sequence have been carried out. The peptides investigated are: Boc-betaHLeu-(Ac6c)2-OMe, Boc-Ac6c-betaHLeu-(Ac6c)2-OMe and Boc-betaHVal-(Aib)5-OtBu, together with the C-protected beta-homo-residue HCl.H-betaHVal-OMe. The results indicate that the insertion of a betaH-residue at position 1 or 2 of peptides containing strong helix-inducing, bulky Calpha,alpha-disubstituted amino acid residues does not induce any specific conformational preferences. In the crystal state, most of the NH groups of beta-homo residues of tri- and tetrapeptides are not involved in intramolecular hydrogen bonds, thus failing to achieve helical structures similar to those of peptides exclusively constituted of Calpha,alpha-disubstituted amino acid residues. However, by repeating the structural motifs observed in the molecules investigated, a beta-pleated sheet secondary structure, and a new helical structure, named (14/15)-helix, were generated, corresponding to calculated minimum-energy conformations. Our findings, as well as literature data, strongly indicate that conformations of betaH-residues, with the micro torsion angle equal to -60 degrees, are very unlikely.

Alkylation↗
PubMed2006

IGF regulation of neutral amino acid transport in the BeWo choriocarcinoma cell line (b30 clone): evidence for MAP kinase-dependent and MAP kinase-independent mechanisms.

OBJECTIVE: IGF-1 and IGF-1 receptors are major determinants of fetal growth and are expressed primarily on the maternal-facing surface of the syncytiotrophoblast cell membrane in the human placenta. IGF-1 regulates fetal growth, in part, by regulating amino acid transport across the placenta. The objective of these studies was to study the role of IGF-1 and its signaling pathway in regulating neutral amino acid transport in a human trophoblast cell culture model. DESIGN: The regulation of neutral amino acid transport by IGF-1 was studied in cultured BeWo(b30) choriocarcinoma cells using the non-metabolizing amino acid analog, [(3)H]-alpha-aminoisobutyric acid (AIB). Transport in the absence of Na was used to distinguish system L from total AIB transport. Similarly, Na-dependent transport in the presence of excess methyl-AIB (MeAIB) permitted discrimination of systems A (MeAIB-sensitive) and ASC (MeAIB-insensitive). Specific inhibitors of intracellular signaling pathways were then used to determine the signaling pathway utilized by IGFs to regulate each amino acid transport system. Specificity of inhibition was assessed using specific markers of p70 S6 kinase activity and MAP kinase activation. RESULTS: Maximal stimulating concentrations of IGF-I (100 ng/ml) stimulated AIB transport by 30-40% exclusively through system A. Wortmannin (100 nM), an inhibitor of PI-3-kinase activity, inhibited all IGF-I-stimulated transport. Rapamycin (100 ng/ml), an inhibitor of p70 S6 kinase, and bisindolylmaleimide, an inhibitor of protein kinase C (PKC), had no effect. PD-098059 (50 miccroM), an inhibitor of MAP kinase activation, inhibited 20-30% of basal AIB transport but did not inhibit IGF-I-stimulated transport under the conditions studied. IGF-1 did not increase steady state mRNA levels of the system A transporters, SNAT1 and SNAT2, suggesting IGF-1 stimulates transport via post-transcriptional mechanisms. CONCLUSIONS: These data demonstrate that IGF-I stimulates neutral amino acid transport system A by a PI3-kinase dependent, post-transcriptional pathway in the BeWo(b30) cell line. Additionally, system A activity appear to be sensitive to MAP kinase-dependent pathways not regulated by IGFs.

Amino Acid Transport System A↗
PubMed1996

rBAT is an amino acid exchanger with variable stoichiometry.

The rBAT protein, when expressed in Xenopus oocytes, was previously shown to reproduce the selectivity of the Na(+)-independent neutral and basic amino acid transport system called bo,+. More recently, the capacity of rBAT to generate a transmembrane current was demonstrated when addition of neutral amino acids stimulated the efflux of cations (presumably basic amino acids) in rBAT-injected oocytes. In the present paper, aminoisobutyric acid (AIB), a neutral amino acid analogue, was shown to induce outward currents (efflux of basic amino acids) through rBAT similar to those caused by alanine in terms of affinity, maximal currents and I-V curves. Despite generating similar currents, the AIB transport rate was more than 30 times lower than that of alanine, thus challenging the assumption that rBAT functions as a classical exchanger. Experiments using a cut-open oocyte voltage clamp demonstrated that AIB was capable of stimulating rBAT-mediated currents from either side of the membrane. AIB, like alanine, was able to stimulate the efflux of radiolabeled alanine and arginine while no rBAT-mediated efflux was measurable in the absence of external rBAT substrates. These results demonstrate that (i) the presence of amino acids is required on both sides of the membrane for rBAT to mediate amino acid flux and thus rBAT must be some type of exchanger but (ii) rBAT-mediated amino acid influx is not stoichiometrically related to the efflux. A model of a "double gated pore" is proposed to account for these properties of rBAT, which contravene standard models of exchangers and other transporters.

Alanine↗
PubMed1975

Adaptive regulation of amino acid transport across the cell membrane in avian and mammalian tissues.

The regulation of amino acid transport across the cell membrane by adaptive mechanisms has been studied in a variety of mesenchymal and epithelial cells and tissues of avian and mammalian origin. Changes in transport activity as a function of time under various in vitro conditions (amino acid dependence, active and inhibited protein synthesis) have been evaluated by measurements of initial entry rates with representative amino acids. Results and conclusions based on the adopted experimental approach include the following. (1) An adaptive control mechanism for the transport of neutral amino acids corresponding to the typical substrates of the A mediation is operative in (a) mesenchymal cells (fibroblasts, chondroblasts, osteoblasts and myoblasts) from embryonic tissues of avian (chick embryo) origin and (b) mesenchymal cells from immature rat uterus (fibroblasts and smooth muscle cells) and other mammalian tissues (cardiac cells from newborn mouse and rat heart). (2) Adaptive regulation is restricted to a discrete subgroup of amino acids (L-proline, glycine and the analogue alpha-aminoisobutyric acid) in rat peritoneal macrophages and thymic lymphocytes. (3) Adaptive regulation is absent in erythroid cells (human erythrocytes, rabbit erythrocytes and reticulocytes, avian erythrocytes) which lack the A mediation and are incapable of active gene transcription. (4) Adaptive regulation is absent in the epithelial kidney cortex tissue and possibly absent in the epithelial component of liver tissue from adult rats; it is fully operative in the chick embryo crystalline lens, i.e. an epithelial preparation of embryonic origin. (5) These observations indicate that adaptive control mechanisms of amino acid transport across the cell membrane are quite common among tissues and species and emphasize their broad biological significance in eukaryotes.

Amino Acids↗
PubMed1993

Plasma membrane activities retained after lethal heat shock.

Cultures of Chinese hamster ovary (CHO) cells were examined to determine if heat killing could be attributed to severe damage in the plasma membrane. Three independent transport activities of the plasma membrane were measured. Glucose transport into the cells (measured with the non-metabolizable analogue 3-O-methyl-D-glucose) was stimulated rather than inhibited by heat. Most of the stimulation was found after non-toxic heat doses. Although amino acid transport (measured with the non-metabolizable analogue 2-aminoisobutyric acid) was slightly inhibited by heat, heat-sterilized cells were able to accumulate high intracellular concentrations. Cellular uptake of the nucleoside uridine was unaffected for at least 4 h after heating. In contrast, its incorporation into RNA was immediately inhibited. To further study plasma membrane damage, cells were either heated or treated with drugs which localize to the plasma membrane, ionophore A23187 or amphotericin B. The mode of cell killing by heat was radically different from that of the two drugs: heat-sterilized cells retained a phase-bright morphology and excluded the viability dye trypan blue while drug-killed cells rapidly became phase-dark and absorbed the dye. These results add to a growing list of plasma membrane activities which are retained in heat-sterilized cells, and suggest that the initial thermal damage responsible for cell killing is at an alternate site(s).

Amino Acids↗
PubMed2006

Genetic analysis of the first 4 patients with beta-ureidopropionase deficiency.

beta-Ureidopropionase is the third enzyme of the pyrimidine degradation pathway and it catalyses the irreversible hydrolysis of N-carbamyl-ss-aminoisobutyric acid or N-carbamyl-ss-alanine to beta-aminoisobutyric acid or ss-alanine, ammonia, and CO2. Analysis of the beta-ureidopropionase gene (UPB1) of the first 4 patients presenting with a complete enzyme deficiency, revealed the presence of 2 splice-site mutations (IVS1-2A>G and IVS8-1G>A) and one missense mutation (A85E). RT-PCR analysis of the complete beta-ureidopropionase cDNA suggested that both splice-site mutations lead to a variety of alternative splice variants, with deletions of a single or several exons. The alanine at position 85 was not conserved in other eukaryotic beta-ureidopropionase protein sequences.

Amidohydrolases↗
PubMed1979

Choline uptake into renal phospholipids following renal ischemia in rats.

Renal phospholipid metabolism was studied after ischemia was induced by occlusion of the left renal artery in the rat. There was no change in the rate of cellular [14C]choline uptake after 25 or 60 minutes of ischemia. However, [14C]choline incorporation into phospholipid was two to three times greater in slices from the ischemic kidney than in slices from the contralateral control kidney. The increase occurred after 25 minutes of ischemia plus 15 minutes of reflow, and after 60 minutes of ischemia with or without reflow. When [14C]choline was injected into rats after a 60-minute period of renal ischemia, the rate of incorporation into phospholipid in the ischemic kidney was almost twice that of the control kidney. These results were similar to those of the in vitro experiments. Since virtually all of the cellular phospholipids of the kidney are present in cellular membranes, renal ischemia affects membrane metabolism. The mean distribution ratio of alpha-aminoisobutyric acid in slices of kidneys ischemic for 60 minutes was similar to that of control slices: 4.11 +/- 0.2 (SEM) vs. 4.30 +/- 0.30. The normal uptake of alpha-aminoisobutyric acid indicates that the increased incorporation of choline is associated with functional integrity of the membrane.

Aminoisobutyric Acids↗
PubMed1984

The nutrient supply of the chick visual system: responses to form and light deprivation.

Blood flow and uptake of two nutrient analogues have been studied in the neural retina, pigment epithelium and optic lobes of 5-7-day-old chicks after systemic administration of isotopically labeled compounds. The pigment epithelium accumulated the inert amino acid, alpha-aminoisobutyric acid, 6 times as effectively on a wet weight basis than did the neural retina or optic lobes. However, the neural retina was over 3 times as active in transport of deoxyglucose as was either the pigment epithelium or optic lobe. These differences could not be attributed to variations in blood flow since the penetrance of the freely diffusible antipyrine was low in the neural retina and roughly equal in the vascularized epithelium and optic lobes. Two days of unilateral eyelid suture significantly depressed blood flow and deoxyglucose uptake in the optic lobe innervated by the occluded eye. The neural retina of the eye receiving reduced light input also had a reduced level of deoxyglucose accumulation but the capacity to concentrate alpha-aminoisobutyrate was increased. The corresponding dark-adapted pigment epithelium had a major (48%) increase in the uptake of this inert amino acid. The data show that regional glucose utilization may not be directly proportional to blood flow or glucose diffusion rates. The non-vascularized neural retina possesses an unusually powerful glucose transport mechanism while the pigment epithelium has a high capacity for amino acid accumulation.

Amino Acids↗
PubMed1990

Expression of the mammalian system A neutral amino acid transporter in Xenopus oocytes.

In this report, we demonstrate the expression of the mammalian System A neutral amino acid transporter in Xenopus laevis oocytes following microinjection of mRNA from rat liver, Chinese hamster ovary (CHO) cells, and human placenta. Stage 6 oocytes were injected with poly(A+) mRNA from one of these three sources and incubated for 24 h prior to assaying Na(+)-dependent 2-aminoisobutyric acid transport to monitor the increase in System A activity. The endogenous 2-aminoisobutyric acid uptake rates in oocytes were sufficiently slow so as to provide a low background value that was subtracted to obtain transport rates for the mammalian carrier alone. The degree of expression of the mammalian System A activity in Xenopus oocytes corresponded to the known transport rates in the tissue from which the mRNA was prepared. For example, hepatic mRNA from glucagon-treated rats produced greater System A activity than mRNA from control animals, and the mRNA from the CHO transport mutant cell line alar4-H3.9, which overproduces System A, resulted in higher transport rates than mRNA from the parental cell line (CHO-K1). Fractionation of total mRNA poly(A+) by nondenaturing agarose gel electrophoresis revealed transport activity associated with a 2.0-2.5-kilobase mRNA fraction common to each of the three tissues tested.

Aminoisobutyric Acids↗
PubMed1988

Effect of chloroquine on membrane permeability in yeast--release of cellular coproporphyrin.

1. The influx and efflux of labelled substances with and without chloroquine was studied in yeast cells. 2. The uptake of delta-aminolevulinic acid by Saccharomyces cerevisiae is characterized by a KT of 3-4 mM and Jmax of 1.0-1.2 mumol min-1 g dry weight-1. 3. A method for loading yeast with labelled coproporphyrin is suggested. 4. The uptake of sorbitol and coproporphyrin was slightly stimulated, while the uptake of 6-deoxyglucose was slightly, that of 2-aminoisobutyric acid and leucine strongly inhibited by chloroquine. 5. The efflux of coproporphyrin, 2-aminoisobutyric acid and sorbitol was stimulated while that of leucine was not influenced by chloroquine. 6. The result showed that chloroquine influenced directly but nonspecifically the membrane permeability, apparently mainly that of the vacuolar membrane.

Aminoisobutyric Acids↗
PubMed1987

Glucose and amino acid uptake by exercising muscles in vivo: effect of insulin, fiber population, and denervation.

The first series of experiments was performed on control rats. The animals were injected iv with trace amounts of 2-deoxy-D-[1-14C]glucose [( 14C]DG) or alpha-[1-14C] aminoisobutyric acid with or without 0.1 U insulin/rat, and calf muscles of the right hindlimb were electrically stimulated to induce 1 contraction/sec. The exercise was discontinued 25 min after the injection, and cellular uptakes of DG or aminoisobutyric acid by soleus, plantaris, and gastrocnemius muscles in disintegrations per min/mg dry tissue weight were determined. The results of these experiments demonstrated that basal, insulin-induced, and exercise-induced uptakes of glucose and amino acids by muscles are dependent on muscle fiber population, and the stimulatory effects of exercise and insulin on soleus and plantaris muscles, but not gastrocnemius muscles, are synergistic rather than merely additive when both stimuli act together. The second series of experiments was performed in the same manner as the first series, except that the right hindlimb of each rat was denervated 3 days before the experiment. Cellular DG uptakes were determined in soleus, plantaris, and gastrocnemius muscles of the left (sham) hindlimb, which was always resting, and in corresponding muscles of the right (denervated) limb, which was either resting or exercising. In the resting state, the denervated soleus muscle had normal basal DG uptake, but, unlike sham soleus muscle, did not respond to insulin stimulation. Denervated plantaris and gastrocnemius muscles exhibited 264% and 150% elevations in basal glucose uptake, respectively, compared with corresponding sham muscles. The latter two denervated muscles did respond to insulin, but the hormone-induced increments in DG uptake were 68% and 45%, respectively, lower than in corresponding contralateral sham muscles. Under basal conditions, exercise had no effect on DG uptake by the denervated soleus muscle, but increased uptake by denervated plantaris and gastrocnemius muscles 207% and 778%, respectively. Stimulation of exercising denervated muscles with exogenous insulin did not increase DG uptake by any muscle above the level observed with exercise alone. The results show that manifestations of insulin resistance in denervated muscles depend on muscle fiber population; denervated muscles, which still retain some ability to respond to insulin, increase DG uptake during exercise, whereas such an increase is absent in denervated muscles completely unresponsive to insulin; and exercise does not improve or restore the ability of denervated muscles to respond to insulin.

Aminoisobutyric Acids↗
PubMed1995

Effects of benfluorex metabolites on membrane fluidity and insulin-related processes.

As little work has dealt with the antihyperglycemic property of benfluorex at the hepatocyte level, we studied the effects of its main metabolites, S422 and S1475, on membrane fluidity and on insulin binding, internalization and action in healthy rat hepatocytes. Both metabolites were effective fluidizing agents. Neither one affected insulin binding. Only S422 favored the bound insulin-receptor internalization process. The metabolites produced no change in basal alpha-aminoisobutyric acid uptake. Only S422 promoted the insulin-stimulated alpha-aminoisobutyric acid uptake in a dose-dependent way. Therefore, our study demonstrated that: (i) the effects of S422 on insulin-related processes in isolated hepatocytes were direct, specific and not due to any membrane fluidizing mechanism; (ii) S422 improved hepatocyte response to insulin at a post-binding level. These results in vitro give an additional explanation, at the cellular level, of the benefit of benfluorex treatment for non insulin-dependent diabetes patients.

Animals↗
PubMed1983

Transport of neutral amino acids by adult articular cartilage.

The transport of three neutral amino acids: alanine, serine and leucine and the non-metabolizable analogue aminoisobutyric acid (AIB) was characterized in bovine articular cartilage slices. Alanine and serine were strongly concentrated intracellularly by a factor of 4.5 and 5 respectively and were transported in a sodium dependent and pH sensitive manner. AIB was concentrated by a factor of 1.8 with respect to the extracellular medium and was also transported in a sodium dependent and pH sensitive manner. Leucine was concentrated weakly across the chondrocyte membrane (X1.1) and its transport was independent of sodium ion concentration and the lowering of the extracellular pH. The apparent Km of transport of alanine, serine, leucine and AIB was determined to be 0.46 mM, 0.69 mM, 0.93 mM and 0.66 mM respectively and the values of Vmax were 35.8, 52.1, 14.8 and 6.8 pmol/min per mg respectively. It was shown that both alanine and serine were transported predominantly by system ASC (63.8% and 67.4% respectively) with a small percentage of transport occurring via system A. AIB was transported mainly by system A (76.0%) and leucine was transported equally via systems L (48.4%) and ASC (45.0%).

Alanine↗