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PubMed1986

Development and characterization of pantothenic acid transport in brain.

In vitro, the transport of [3H]pantothenic acid into and from rabbit brain slices was studied. In newborn rabbits and throughout development, forebrain and cerebellar slices were able to accumulate and phosphorylate [3H]pantothenic acid comparably to slices from adults. The accumulation and phosphorylation of [3H]pantothenic acid by adult forebrain slices were not decreased by substitution of LiCl for NaCl in the artificial CSF or by addition of short-chain fuels (e.g., 5 mM pyruvate or acetoacetate) to the medium. However, probenecid and ouabain (both 1 mM) and medium-chain fatty acids (e.g., 0.1 mM octanoate, nonanoate, and decanoate) profoundly inhibited [3H]pantothenic acid accumulation by forebrain slices but not intracellular phosphorylation and conversion to [3H]CoA. There in vitro results suggest that brain slices accumulate pantothenic acid by a saturable system (probably facilitated diffusion) that is sensitive to inhibition by probenecid and medium-chain fatty acids.

Animals↗
PubMed1989

Sodium-dependent uptake of nucleosides by dissociated brain cells from the rat.

Sodium-dependent 3H-labeled nucleoside transport was studied using a mixed population of dissociated brain cells from adult rats. The accumulation of [3H]adenosine during brief (15-s) incubation periods was significantly greater in the presence of 110 mM Na+ than in its absence. This occurred at substrate concentrations that ranged from 0.25 to 100 microM. Similar findings were observed for the rapid accumulation of [3H]uridine. Kinetically, the rapid accumulation of [3H]adenosine in both the absence and the presence of Na+ was best described by a two-component system. In the presence of Na+, the KT and Vmax values for the high-affinity affinity component were 0.9 microM and 8.9 pmol/mg of protein/15 s, and those for the low-affinity component were 313 microM and 3,428 pmol/mg of protein/15 s, respectively. In the absence of Na+, the KT value for the high-affinity component was significantly higher (1.8 microM). [3H]Uridine accumulation was best described kinetically by a one-component system that in the presence of Na+ had KT and Vmax values of 1.0 mM and 2.6 nmol/mg of protein/15 s, respectively. As was found for [3H]adenosine, in the absence of Na+, the KT value was significantly higher (1.8 mM). The sodium-dependent transport of [3H]adenosine was inhibitable by ouabain and 2,4-dinitrophenol. Of the three nucleoside transport inhibitors tested, only nitrobenzylthioninosine demonstrated high affinity and selectivity in blocking the sodium component. Thus, high-affinity sodium-dependent nucleoside transport systems, in addition to facilitated diffusion systems, exist on brain cells from adult rats.

Adenosine↗
PubMed1992

Biotin transport in primary culture of astrocytes: effect of biotin deficiency.

The uptake of radioactive biotin has been studied in glial cell cultures from dissociated cerebral hemispheres of newborn rats. We describe saturable kinetics for the biotin uptake at biotin concentrations of less than 60 nM and linear at greater than 60 nM. The uptake appeared temperature sensitive, Na+ independent, nonsensitive to valinomycin, and not affected by metabolic inhibitors such as sodium fluoride or azide. Lipoic acid and biocytin were effective in inhibiting the biotin uptake. These findings are consistent with biotin uptake by the primary culture of astrocytes as a process of facilitated diffusion. Moreover, biotin uptake in astrocytes grown in biotin-restricted conditions was significantly higher compared with the control. This increase appeared mediated through a pronounced increase (10-fold) in the Vmax of the biotin uptake without any change in the apparent Km.

Animals↗
PubMed1985

The mechanism of adenosine release from hypoxic rat liver cells.

Uptake of [14C]-adenosine into freshly dispersed rat hepatocytes was inhibited 44% by dipyridamole (50 microM) and 60% by nitrobenzylthioinosine (NBTI, 20 microM). The results are consistent with the known ability of these drugs to inhibit adenosine transport in other cell types. The nucleotide analogue, alpha, beta-methylene adenosine diphosphate (AOPCP, 50 microM), inhibited by 84% the degradation of exogenous 5' AMP that occurred rapidly when this substrate alone was presented to isolated hepatocytes. This confirms the ecto-5'-nucleotidase inhibitory properties of this analogue in isolated hepatocytes. During hypoxic incubation, isolated hepatocytes released adenosine, which accumulated in the extracellular volume. Dipyridamole and NBTI each markedly attenuated this extracellular adenosine accumulation. In contrast, AOPCP had no inhibitory effect on net hypoxic adenosine release. It is concluded that hypoxic rat hepatocytes produce adenosine intracellularly and that this adenosine is released via facilitated diffusion to the extracellular space, based on the inhibition observed with the transport inhibitors. The plasma membrane enzyme ecto-5'-nucleotidase does not appear to participate in hypoxic adenosine release from these cells as indicated by the lack of effect of the nucleotidase inhibitor, AOPCP.

Adenosine↗
PubMed1988

Blockade of the inotropic effect of Bay K 8644 by cytochalasin-B and phloretin.

1. The positive inotropic effect in rabbit atria and papillary muscles of Bay K 8644 is blocked by cytochalasin-B (Cyto-B) and phloretin, two compounds known to block the facilitated diffusion of glucose. These compounds do not change the concentration-response curve of calcium. 2. Cyto-B is more potent in atria than in papillary muscles, 10(-7) M having a maximal effect in atria whereas 2 x 10(-5) M was required for a maximal effect in papillary muscles. Phloretin was fully effective at 10(-4) M, the only concentration tested. 3. The inotropic effect of Bay K 8644 was virtually abolished in atria bathed in a glucose-free medium or one containing 5 mM pyruvate. The contractile response to Bay K 8644 of papillary muscles was not changed significantly in glucose-free or in pyruvate-containing medium. 4. Cyto-B (2 x 10(-5) M) caused a slight but significant increase in the KD for the binding of nitrendipine to a crude sarcolemnal preparation from rabbit ventricles. The Bmax was unchanged. 5. These results may best be explained by the hypothesis that there is a metabolic requirement for the inotropic effect of Bay K 8644.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed2006

Flow distribution analysis by helical scanning in polysulfone hemodialyzers: effects of fiber structure and design on flow patterns and solute clearances.

The efficiency of a hemodialyzer is largely dependent on its ability to facilitate diffusion, as this is the main mechanism by which small solutes are removed. The diffusion process can be impaired if there is a mismatch between blood and dialysate flow distribution in the dialyzer. The objective of the paper was to study the impact of different fiber bundle configurations on blood and dialysate flow distribution and urea clearances. The Optiflux 200 NR hemodialyzer was studied and the standard F 80 A hemodialyzer was used as a control for the study. Six dialyzers of each type were studied in vitro in the radiology department utilizing a new generation of helical computed tomography (CT) scan following contrast medium injection into the blood and dialysate compartment. Dynamic sequential imaging of longitudinal sections of the dialyzer was undertaken to detect flow distribution, average and peak velocities, and calculate wall shear rates. Six patients were dialyzed with 2 different dialyzers in random consecutive sequence. In these patients, 2 consecutive dialyses were carried out with identical operational parameters (Qb = 300 mL/min, Qd = 500 mL/min). In each session, blood and dialysate side urea clearances were measured at 30 and 150 min of treatment. Macroscopic and densitometrical analysis revealed that flow distribution was most homogeneous in the dialyzer with a new bundle configuration. Significantly increased urea clearances (p < 0.001) were seen with the Optiflux dialyzer compared with the standard dialyzer. In conclusion, more homogeneous dialysate blood and dialysate flow distribution and improved small solute clearances can be achieved by modifying the configuration of the filter bundle. These effects are achieved probably as a result of reduced blood to dialysate mismatch with reduction of flow channeling. The used radiological technique allows detailed flow distribution analysis and has the potential for testing future modifications to dialyzer design.

Blood Flow Velocity↗
PubMed2003

High capacity xylose transport in Candida intermedia PYCC 4715.

Xylose-utilising yeasts were screened to identify strains with high xylose transport capacity. Among the fastest-growing strains in xylose medium, Candida intermedia PYCC 4715 showed the highest xylose transport capacity. Maximal specific growth rate was the same in glucose and xylose media (mu(max)=0.5 h-1, 30 degrees C). Xylose transport showed biphasic kinetics when cells were grown in either xylose- or glucose-limited culture. The high-affinity xylose/proton symport system (Km = 0.2 mM, Vmax = 7.5 mmol h-1 g-1) was more repressed by glucose than by xylose. The less specific low-affinity transport system (K = 50 mM, Vmax = 11 mmol h-1 g-1) appeared to operate through a facilitated-diffusion mechanism and was expressed constitutively. Inhibition experiments showed that glucose is a substrate of both xylose transport systems.

Biological Transport, Active↗
PubMed2000

Alternative pathways of glucose transport in Prevotella bryantii B(1)4(1).

Prevotella bryantii B(1)4 grew faster on glucose than mannose (0.70 versus 0.45 h(-1)), but these sugars were used simultaneously rather than diauxically. 2-deoxy-glucose (2DG) decreased the growth rate of cells that were provided with either glucose or mannose, but 2DG did not completely prevent growth. Cells grown on glucose or mannose transported both (14)C-glucose and (14)C-mannose, but cells grown on glucose had over three-fold higher rates of (14)C-glucose transport than cells grown on mannose. The (14)C-mannose transport rates of glucose- and mannose-grown cells were similar. Woolf-Augustinsson-Hofstee plots were not linear, and it appeared that the glucose/mannose/2DG carrier acted as a facilitated diffusion system at high substrate concentrations. When cultures were grown on nitrogen-deficient (excess sugar) medium, isolates had three-fold lower (14)C-glucose transport, but the (14)C-mannose transport did not change significantly. (14)C-glucose and (14)C-mannose transport rates could be inhibited by 2DG and either mannose or glucose, respectively. The (14)C-glucose transport of mannose-grown cells was inhibited more strongly by mannose and 2DG than those grown on glucose. Cells grown on glucose or mannose had similar ATP-dependent glucokinase activity, and 2DG was a competitive inhibitor (K(i)=0.75 mM). Thin layer chromatography indicated that cell extracts also had ATP-dependent mannose phosphorylation, but only a small amount of phosphorylated 2DG was detected. Glucose, mannose or 2DG were not phosphorylated in the presence of PEP. Based on these results, it appeared that P. bryantii B(1)4 had: (1) two mechanisms of glucose transport, a constitutive glucose/mannose/2DG carrier and an alternative glucose carrier that was regulated by glucose availability, (2) an ATP-dependent glucokinase that was competitively inhibited by 2DG but was unable to phosphorylate 2DG at a rapid rate, and (3) virtually no PEP-dependent glucose, mannose or 2DG phosphorylation activities.

Biological Transport↗
PubMed1981

Lead poisoning and the blood-brain barrier.

Lead exposure may produce varying degrees of neuropsychiatric manifestations from discrete phenomena, quite often seen in children and as an occupational disease, to the rate fulminant lead encephalopathy. It was determined whether or not damage of the blood-brain barrier permeability in adult rats, as has been demonstrated in neonatal animals exposed to lead, could also play a role. Massive lead exposure did not induce any change in the transfer (facilitated diffusion) of phenylalanine any tyrosine measured by means of the indicator dilution technique. Ultrastructural examination, after application of horseradish peroxidase, did not reveal any pathological changes in the permeability to the tracer. It is concluded that in adult rats, in contrast to neonatal animals, the observed pathological signs clearly seen in the chronically exposed animals must be ascribed to a noxious influence of lead on the extravascular side of the blood-brain barrier.

Animals↗
PubMed2006

Availability of fluoride from meals given to kindergarten children in Brazil.

OBJECTIVES: The aim of this study was to evaluate the amount of fluoride supplied daily in the meals given in 44 public kindergarten schools in Bauru, Brazil. In addition, the fluoride concentration of water supplies and its impact on the amount of fluoride found in the meal samples were also investigated. METHODS: Meal samples and water were collected during 2 weeks (10 working days) in public kindergarten schools. Samples of meals were homogenized with known volumes of deionized water. Fluoride present in meal samples was analyzed with the ion-specific electrode (Orion 9609), after hexamethyldisilazane-facilitated diffusion. Fluoride in water samples was analyzed with the same electrode, after buffering with TISAB II. All the analyses were made in duplicate. RESULTS: Fluoride analyzed (mean+/-SD) was 0.50+/-0.20 microg/ml, ranging from nondetectable to 1.42 microg/ml for water samples (n=424) and 0.067+/-0.059 mg, ranging from 0.007 to 0.580 mg for meal samples (n=431). A weak but significant correlation was observed between the amount of fluoride in meals and fluoride concentration in water supplies (r=0.139, P=0.0042). CONCLUSIONS: Despite the seemingly small role played by school meals in the total daily fluoride intake, they can contribute to the total fluoride intake of children on a chronic basis, when in association with other fluoride products. Additionally, the impact of fluoridated public water supply on the final fluoride concentration of the school meals analyzed may be regarded as low.

Animals↗
PubMed1977

Influence of diabetes on metabolism of vascular smooth muscle.

Glucose utilization and the conversion of glucose to lactate, CO2, glycogen and lipids are decreased in the aorta from diabetic rats and rabbits. In addition the incorporation of amino acid into protein is reduced in diabetic rat aorta. The metabolic changes produced by diabetes are counteracted by insulin treatment, but there is a time lag of about 2 days before the effect of insulin treatment appears. The membrane transport of glucose in smooth muscle is carried out by a specific transport system of the facilitated diffusion type. A rate limiting influence of membrane transport on glucose metabolism is found in bovine mesenteric arteries and rabbit colon smooth muscle. In these preparations the influence of glucose concentrations on glucose metabolism is most pronounced in the range 0-11.1 mmol exhibiting saturation at higher glucose concentrations. Insulin in a high concentration (0.1 U/ml) has acute (less than or equal to 3 h) metabolic effects in vitro on smooth muscle which are qualitatively similar to those in skeletal muscle, but are weaker and appear later. The threshold concentration for the acute metabolic effects of insulin on smooth muscle in vitro is 10-100 times above the physiological levels, indicating a low acute sensitivity to insulin.

Animals↗
PubMed1989

Lactate transport in isolated mouse muscles studied with a tracer technique--kinetics, stereospecificity, pH dependency and maximal capacity.

Lactate transport across the sarcolemma of isolated mouse muscles was studied with a 14C tracer technique. The cellular tracer uptake could be inhibited by unlabelled L-lactate (and pyruvate) and to a lesser extent by D-lactase. The stereospecific fraction had a Km of 3.5 mM, and made up 50% of the total transport. The tracer uptake was unaffected by 0.05 mM DIDS and 0.2 mM amiloride, but was inhibited by cinnamate (Ki = 8 mM) and PCMBS (Ki = 0.8 mM). With high concentrations of the latter inhibitor compounds or with high concentrations of unlabelled L-lactate, the tracer uptake was inhibited 80%, which indicates that the main part of the transport involves facilitated diffusion. The remaining fraction (20%) was non-saturable, reduced at high pH, and could not be inhibited; it is probably mediated by diffusion of undissociated lactic acid. Lactate transport was pH-dependent, which is consistent with a lactate-H+ symport. The maximal transport capacity, as calculated from the pH changes measured with pH-sensitive micro-electrodes while the lactate gradient was 30 mM, was 11.8 mmol kg-1 min-1 (pH 6.2).

4-Chloromercuribenzenesulfonate↗
PubMed1975

INtestinal sugar transport: studies with isolated plasma membranes.

The luminal and the contraluminal plasma membranes of small intestinal epithelial cells were isolated separately and in the form of membrane vesciles. In the luminal membrane distinct transport systems exist for D-fructose and for D-glucose and glucalogs as evidenced by stereospecificity for the substrate, counter- and cotransport phenomena, and inhibitor specificity. D-Glucose is taken up against a concentration gradient when energy is supplied in the form of an electrochemical Na+ gradient between medium and intravesicular membrane space. The contraluminal membrane also possesses a glucose transport system. However, it is sensitive to different inhibitors than the glucose transport in the luminal membrane. D-Glucose uptake by the contraluminal membrane is not enhanced by Na+ gradients and can be characterized as "facilitated diffusion." The studies with the isolated plasma membranes allow a description of transcellular D-glucose transport in terms of the translocation events across single membranes.

Animals↗
PubMed1981

Transport properties of intestinal basolateral membranes.

Techniques for the isolation and study of basolateral membrane vesicles from the intestinal epithelium have afforded new insights into the mechanisms of intestinal absorption. First, we have confirmed the hypothesis that the second stage of glucose transport involves facilitated diffusion. Second, we have shown that the major system for translocation of neutral amino acids across the basolateral membrane is the classical "L" system. Third, we have established that basolateral membranes contain sodium-dependent transport systems that may be useful in the supply of essential amino acids to the epithelium from the blood. And, finally, our studies of the basolateral (Na + K)-ATPase have clarified the role of this enzyme in sodium absorption.

4-Chloromercuribenzenesulfonate↗
PubMed1984

Chemical properties, distribution, and physiology of plant and algal carbonic anhydrases.

Plant carbonic anhydrases (CAs) have a range of molecular weights (MW). Among flowering plants, dicotyledons with C3 photosynthesis have two isoenzymes of 140-250K each with 6 subunits, while monocotyledons have two isoenzymes of 42-45K. Plant and animal CAs have a similar amino acid content, subunit size and zinc content, suggesting they are homologous proteins, although the higher plant CAs have no esterase activity and are not strongly inhibited by sulfonamides. Algal CAs vary widely in MW and some are highly sensitive to sulfonamides like the animal enzymes. The two plant isoenzymes, from the chloroplast and cytosol, can be separated by gradient polyacrylamide gel electrophoresis and subsequently visualized by enzymic H+ ion production. In plants, CAs probably facilitate diffusion of CO2 to the site of photosynthetic fixation; they may also have a role in pH regulation, in the use of bicarbonate by aquatic plants and in concentrating inorganic carbon within the chloroplast.

Carbonic Anhydrases↗
PubMed2000

Coupling between Na+, sugar, and water transport across the intestine.

Water is absorbed across the small intestine in the absence of external driving forces. However, it has been established that water transport is secondary to active sodium transport. In the upper intestine both sodium and water absorption are largely dependent on the presence of D-glucose. The link between active sodium transport and glucose is the coupled transport of sodium and glucose across the brush border membrane of enterocytes by the Na+/glucose cotransporter (SGLT1). Na+ that enters the cells with glucose is pumped out towards the blood by 3Na+/2K+ pumps on the basolateral membrane, and glucose passes out across the basolateral membrane by facilitated diffusion, the net result being that glucose and sodium are transported across the epithelium. The coupling between Na+, glucose, and water transport is less well understood. It is commonly thought that Na+ transport increases the local osmotic pressure in the lateral intercellular spaces, and that this in turn generates osmotic water flow across the epithelium. Recent work suggests a more direct link between Na+, glucose, and water transport; that is, water is cotransported along with Na+ and sugar through SGLT1. Here we review the evidence for Na+/glucose/water cotransport.

Animals↗
PubMed1977

On the molecular biology of the vitreous in the aphakic eye.

The vitreous of aphakic and phakic eyes was assayed for hyaluronic acid. Intracapsular cataract extraction was regularly followed by decreased hyaluronic acid concentration and studies on the distribution of hyaluronic acid within the vitreous cavity showed that removal of the lens facilitates diffusion of hyaluronic acid into the anterior chamber. Based on these studies certain postoperative features of the aphakic eye are discussed. The instability of the vitreous gel, as reflected by rupture of the anterior vitreous face and an increased incidence of vitreous detachment supports the hypothesis of hyaluronic acid as a stabilizer of the gel. The decreased adhesion of the neural retina in the aphakic eye might be caused by the loss of hyaluronic acid. Normally the high concentration of hyaluronic acid adjacent to the retina contributes to the resistance of bulk flow of fluid into the subretinal space making it possible for the active transport system of the pigment epithelium to maintain a pressure drop across the neural retina. It is anticipated that saccades create currents in the fluid vitreous close to the eye wall after vitreous detachment. Model experiments showed that these currents were greatly enhanced, when the concentration of hyaluronic acid was lowered.

Aged↗
PubMed1987

The effect of a metabolic inhibitor upon the properties of the cerebral vasculature during a whole-head saline perfusion of the rat.

The effect of the metabolic inhibitor 2,4-dinitrophenol (DNP) has been assessed during a simple in situ Ringer solution perfusion of the rat brain. The preparation was perfused, with or without the addition of DNP, for periods ranging up to 30 min. Following this pre-test perfusion, both the vascular permeability and cerebral perfusate flow were assessed. In the absence of DNP significant barrier disruption had taken place by 10 min and the flow rates showed greater fluctuations with time. In the presence of DNP, however, perfusate flow remained constant and the blood-brain barrier remained intact to [14C]mannitol for at least 10 min, but subsequently the flow rate dropped and the barrier began to show evidence of disruption. The unbound visual marker, Evans Blue, was apparently excluded from all regions other than those that are known to lack a blood-brain barrier. The water content of the brain showed no significant increase until 20 min. Patency of the capillaries was demonstrated by direct visualization of the cerebral vasculature with an Indian ink-gelatin mixture and in some animals there was evidence of incomplete filling following 30 min of perfusion. It is concluded that the use of DNP in the perfusate provides a useful preparation for the short-term study of passive properties of the blood-brain barrier, such as carrier-facilitated diffusion, as well as mechanisms of barrier opening.

2,4-Dinitrophenol↗