Lactate Ions Research Articles

PH compartmented w/o/w multiple emulsion: a diffusion study

In order to develop w/o/w emulsions characterized by two separate aqueous phases of different pH, a preliminary study was carried out to obtain a better insight into the possible diffusion processes taking place between an inner acidic aqueous phase and an external phase of higher pH (pH≈6). In fact, such systems could be of great interest for pharmaceutical use. For this purpose, a model emulsion was formulated. The study of pH and conductivity showed that acidic species transport take place between the two aqueous compartments. The three main release mechanisms that might be responsible for this passage across the oil phase were investigated: breakdown of oil globules, facilitated transport by surfactant micelles across the oil phase or by Fickian diffusion. It appears that this last mechanism was involved. In order to control this diffusion process, an alkaline species, octadecylamine was introduced in the oil phase. This compound could form an ion pair with the lactate ion at the interface of the external aqueous phase and the oil phase, thus, limiting the acidification of the external aqueous phase.

Mouse MCT3 gene is expressed preferentially in retinal pigment and choroid plexus epithelia.

Monocarboxylate transporters (MCTs) are a family of highly homologous membrane proteins that mediate the 1:1 transport of a proton and a lactate ion. In chicken, MCT3 is preferentially expressed in the retinal pigment epithelium (RPE). We have isolated the mouse MCT3 cDNA and gene and characterized the pattern of tissue expression. MCT3 is a single copy gene with a 1.8-kb transcript that encodes a protein with a predicted molecular mass of 51.5 kDa. Based on Northern hybridization analysis, MCT3 transcript was expressed in only two tissues: RPE and choroid plexus epithelium (CPE). The choroid plexus forms a barrier between the cerebrospinal fluid and fenestrated capillaries, similar to the organization of the RPE and choroidal vessels. Immunohistochemical staining demonstrated that MCT3 was restricted to the basolateral membranes of both epithelia but was more abundant in RPE than CPE. Differences in the level of protein expression were confirmed by Western blot analysis. The cloning of MCT3 identifies a specific transporter that could regulate lactate levels in fluid-bathing neuronal tissues.

Determination of transport kinetics of chick MCT3 monocarboxylate transporter from retinal pigment epithelium by expression in genetically modified yeast.

Monocarboxylate transporters (MCTs) comprise a group of highly homologous proteins that reside in the plasma membrane of almost all cells and which mediate the 1:1 electroneutral transport of a proton and a lactate ion. The isoform MCT3 is restricted to the basal membrane of the retinal pigment epithelium where it regulates lactate levels in the neural retina. Kinetic analysis of this transporter poses formidable difficulties due to the presence of multiple lactate transporters and their complex interaction with MCTs in adjacent cells. To circumvent these problems, we expressed both the MCT3 gene and a green fluorescent protein-tagged MCT3 construct in Saccharomyces cerevisiae. Since L-lactate metabolism in yeast depends on the CYB2 gene, we disrupted CYB2 to study the MCT3 transporter activity free from the complications of metabolism. Under these conditions L-lactate uptake varied inversely with pH, greater uptake being associated with lower pH. Whereas the V(max) was invariant, the K(m) increased severalfold as the pH rose from 6 to 8. In addition, MCT3 was highly resistant to a number of "classical" inhibitors of lactate transport. Last, studies with diethyl pyrocarbonate and p-chloromercuribenzenesulfonate set limitations on the locus of potential residues involved in the critical site of lactate translocation.

Free radical scavenging and antioxidant effects of lactate ion: an in vitro study.

Divergent literature data are found concerning the effect of lactate on free radical production during exercise. To clarify this point, we tested the pro- or antioxidant effect of lactate ion in vitro at different concentrations using three methods: 1) electron paramagnetic resonance (EPR) was used to study the scavenging ability of lactate toward the superoxide aion (O(2)(-).) and hydroxyl radical (.OH); 2) linoleic acid micelles were employed to investigate the lipid radical scavenging capacity of lactate; and 3) primary rat hepatocyte culture was used to study the inhibition of membrane lipid peroxidation by lactate. EPR experiments exhibited scavenging activities of lactate toward both O(2)(-). and.OH; lactate was also able to inhibit lipid peroxidation of hepatocyte culture. Both effects of lactate were concentration dependent. However, no inhibition of lipid peroxidation by lactate was observed in the micelle model. These results suggested that lactate ion may prevent lipid peroxidation by scavenging free radicals such as O(2)(-). and.OH but not lipid radicals. Thus lactate ion might be considered as a potential antioxidant agent.

Arterio-venous differences of blood acid-base status and plasma sodium caused by intense bicycling.

After intense exercise muscle may give off hydrogen ions independently of lactate, perhaps by a mechanism involving sodium ions. To examine this possibility further five healthy young men cycled for 2 min to exhaustion. Blood was drawn from catheters in the femoral artery and vein during exercise and at 1-h intervals after exercise. The blood samples were analysed for pH, blood gases, lactate, haemoglobin, and plasma proteins and electrolytes. Base deficit was calculated directly without using common approximations. The leg blood flow was also measured, thus allowing calculations of the leg's exchange of metabolites. The arterial blood lactate concentration rose to 14.2 +/- 1.0 mmol L-1, the plasma pH fell to 7. 18 +/- 0.02, and the base deficit rose 22% more than the blood lactate concentration did. The femoral-venous minus arterial differences peaked at 1.8 +/- 0.2 mmol L-1 (lactate), -0.24 +/- 0.01 (pH), and 4.5 +/- 0.4 mmol L-1 (base deficit), and -2.5 +/- 0.7 mmol L-1 (plasma sodium concentration corrected for volume changes). Thus, near the end of the exercise and for the first 10 min of the recovery period the leg gave off more hydrogen ions than lactate ions to the blood, and sodium left plasma in proportion to the extra hydrogen ions appearing. The leg's integrated excess release of hydrogen ions of 0.88 +/- 0.45 mmol kg-1 body mass was 67% of the integrated lactate release. Base deficit calculated by the traditional approximate equations underestimated the true value, but the error was less than 10%. We conclude that intense exercise and lactic acidosis may lead to a muscle release of hydrogen ions independent of lactate release, possibly by a Na+,H+ exchange. Hydrogen ions were largely buffered in the red blood cells.

M-wave changes after high- and low-frequency electrically induced fatigue in different muscles.

The mechanisms of fatigue-induced changes in evoked compound muscle action potential (M-waves) are not well documented. In rats, isolated fast-glycolytic (tibialis anterior, or TA) and slow-oxidative muscles (soleus, or SOL) were stimulated repetitively at a low (10-Hz) or high (80-Hz) rate. Decreased amplitude and prolonged duration of M-waves were only significant after high-frequency fatigue in TA and SOL muscles; that is, in the conditions in which an influx of calcium was measured. On the other hand, maximal force failure and maximal lactic acid increase in the bath medium occurred in TA muscle after high- and low-frequency fatigue trials. Postfatigue increase in potassium concentration occurred in all circumstances. Thus, M-wave alterations depend mainly on the stimulation paradigm and not on the muscle type, and cannot be used as a marker of changes in intracellular potassium or lactate ions. This amplifies understanding of the significance of M-wave changes in human exercise protocols.

Disparate effects of three types of extracellular acidosis on left ventricular function.

Effects of acidosis on muscle contractile function have been studied extensively. However, the relative effects of different types of extracellular acidosis on left ventricular (LV) contractile function, especially the temporal features of contraction, have not been investigated in a single model. We constituted perfusion buffers of identical ionic composition, including Ca2+ concentration ([Ca2+]), to mimic physiological control condition (pH 7.40) and three types of acidosis with pH of 7.03: inorganic (IA), respiratory (RA), and lactic (LA). Isolated rabbit hearts (n = 9) were perfused with acidotic buffers chosen at random, each preceded by the control buffer. Under steady-state conditions, instantaneous LV pressure (Pv) and volume (Vv) were recorded for a range of Vv. The results were as follows. 1) LV passive (end-diastolic) elastance increased with IA and RA. However, this increase may not be a direct effect of acidosis; it can be explained on the basis of myocardial turgor. 2) Although LV inotropic state (peak active Pv and elastance) was depressed by all three acidotic buffers, the magnitude of inotropic depression was significantly less for LA. 3) Temporal features of Pv were altered differently. Whereas IA and RA reduced time to peak Pv (tmax) and hastened isovolumic relaxation at a common level of LV wall stress, LA significantly increased tmax and retarded relaxation. These results and a model-based interpretation suggest that cooperative feedback (i.e., force-activation interaction) plays an important role in acidosis-induced changes in LV contractile function. Furthermore, it is proposed that LA-induced responses comprise two components, one due to intracellular acidosis and the other due to pH-independent effects of lactate ions.

In Situ Infrared Spectroscopic Studies of Adsorption of Lactic Acid and Related Compounds on the TiO2 and CdS Semiconductor Photocatalyst Surfaces from Aqueous Solutions

Abstract In situ attenuated total reflectance infrared (ATR-IR) spectroscopy has been used to determine the structure of adsorbed lactic acid (LA) compounds on the surfaces of titanium dioxide (TiO2) and cadmium sulfide (CdS) photocatalysts. LA was found to chelate onto the TiO2 in a b identate fashion, while in the CdS case, LA was adsorbed electrostatically as the lactate ion. These differing surface interactions are likely to determine the selectivity in the photo-products of LA.

Effect of arterial occlusion on responses of group III and IV afferents to dynamic exercise.

Our laboratory has shown previously that a low level of dynamic exercise induced by electrical stimulation of the mesencephalic locomotor region (MLR) stimulated group III and IV muscle afferents in decerebrate unanesthetized cats (C. M. Adreani, J. M. Hill, and M. P. Kaufman. J. Appl. Physiol. 83: 1811-1817, 1997). In the present study, we have extended these findings by examining the effect of occluding the arterial supply to the dynamically exercising muscles on the afferents' responses to MLR stimulation. In decerebrate cats, we found that arterial occlusion increased the responsiveness to a low level of dynamic exercise in 44% of the group III and 47% of the group IV afferents tested. Occlusion, compared with the freely perfused state, did not increase the concentrations of either hydrogen ion or lactate ion in the venous effluent from the exercising muscles. We conclude that arterial occlusion caused some unspecified substance to accumulate in the working muscles to increase the sensitivity of equal percentages of group III and IV afferents to dynamic exercise.

Metabolic flux analysis of lactic acid fermentation: effects of pH and lactate ion concentration

A detailed metabolic flux analysis for lactic acid production by Streptococcus lactis has been carried out. A metabolic reaction set was constructed for the metabolism of S. lactis. Fluxes through these reactions were estimated by using accumulation rates of biomass, product and consumption rates of the substrate, which were obtained through experiments. The changes in the flux movement are shown for different pHs and initial lactate concentrations of the medium. The analysis indicated that pH only affected the uptake rates of lactose, whereas lactate ion concentration influenced the movement of flux through the network.

Elucidation of the mechanism of lactic acid growth inhibition and production in batch cultures of Lactobacillus rhamnosus

Batch cultures of Lactobacillus rhamnosus were carried out at different pH values in order to study the limitation of growth and lactic acid production by the hydrogen ion, non-dissociated lactic acid and internal lactate concentrations. The effect of pH between 5 and 6.8 was studied at non-limiting concentrations of glucose; this is more significant for the lactic acid fermentation rate than for the maximum specific growth rate, as shown by the incomplete substrate consumption at lower values of medium pH and by the constant maximum cell mass obtained within the range of pH values studied. To check whether these results were a direct consequence of the different concentrations of the non-dissociated form of lactic acid at different external pH values, specific growth rates and lactic acid productions rates were calculated for each external pH value. The same specific growth rates were observed at the same non-dissociated lactic acid concentrations only at pH values of 5 and 5.5. For higher values of pH (pH > 6) the specific growth rate falls to zero as the non-dissociated lactic acid concentration decreases. This shows that generalisations made from studies performed within very narrow ranges of pH are not valid and that the non-dissociated form of lactic acid is not the only inhibiting species. The internal pH was measured experimentally for each external pH value in order to calculate the internal lactate ion concentration. This form is described to be the inhibitory one. The results obtained confirmed that the specific growth rate reached zero at approximately the same lactate concentration for all the pH values studied.

Stimulation of pulmonary C fibres by lactic acid in rats: contributions of H+ and lactate ions.

1. The contributions of H+ and lactate ions to the stimulation of single pulmonary C fibres by lactic acid were examined in anaesthetized and artificially ventilated rats. 2. Lactic acid injected into the right atrium caused a transient decrease in arterial blood pH (pHa) and a short but intense burst of afferent activities in pulmonary C fibres, whereas sodium lactate had no effect. The fibre activity usually reached a peak within 1-1.5 s, with an onset latency of < 1 s, and returned to the baseline in 5 s. 3. The injection of hydrochloric acid at the same pH as that of lactic acid did not significantly decrease pHa, nor did it stimulate any C fibres studied. 4. Formic acid has a pKa value (the negative logarithm of the dissociation constant) almost identical to that of lactic acid; thus, its injection decreased pHa to the same degree as did the injection of lactic acid. However, the response of C fibres to lactic acid was 134% stronger than that to formic acid. 5. We conclude that H+ is primarily responsible for the activation of pulmonary C fibres by lactic acid, probably through a direct effect of H+ on these afferent endings. The lactate ion, by itself, does not activate C fibres, but it seems to potentiate the stimulatory effect of H+ on these afferents.

Interaction of Alkaline Earth Metal Ions with Acetic and Lactic Acid in Aqueous Solutions Studied by 13C NMR Spectroscopy

Abstract Interaction of alkaline earth metal (magnesium, calcium, strontium and barium) ions with acetic and lactic acid in aqueous media was investigated by 13C NMR spectroscopy. In the acetate systems, signals whose chemical shifts were the averages of those of the free and bound acetate ions were observed. Downfield shifts of the carboxylate carbon signals with increasing metal ion concentration indicated that the acetate ion acted as a monodentate ligand coordinating to the metal ion using the carboxylate group. The metal ion concentration dependence of the peak positions of the methine and carboxylate carbon signals of the lactate ion in the lactate systems suggested that the lactate ion coordinated to a metal ion using the carboxylate and hydroxyl groups. Unique upfield shifts upon complexation in the magnesium lactate systems suggested that the lactate ion coordinated to the magnesium ion from outside the primary hydration sphere. The present results were consistent with the isotope effects of the alkaline earth metals observed in cation exchange chromatography.

Structured Model for Batch Culture Growth ofLactobacillus bulgaricus

Batch fermentation kinetics of Lactobacillus bulgaricus were examined in detail using the methodology of cybernetic modelling. The effect of pH and lactate ion on the activity of the enzyme β-galactosidase was simulated. Cybernetic modelling is mainly used for simulation of growth on multiple substrates. Here, it has been incorporated in a model which simulates pH effects on single-substrate batch growth. The expression of active enzyme is crucial for substrate and growth. It was seen that the effect of lactate ion on the activity of the enzyme was dependant on the system pH. A semi-empirical expression was obtained for the relative amount of active enzyme present in the organism and used to model the enzyme, biomass, substrate and lactic acid concentrations for the batch fermentation. The rate of biomass formation and product formation depended on the amount of enzyme synthesised, which in turn was dependant on the pH value of the system. The model developed simulates the effect of pH and lactate concentration on the expression and degradation of the enzyme. © 1997 SCI.

Diet composition and the performance of high-intensity exercise

The crucial role of muscle glycogen as a fuel during prolonged exercise is well established, and the effects of acute changes in dietary carbohydrate intake on muscle glycogen content and on endurance capacity are equally well known. More recently, it has been recognized that diet can also affect the performance of highintensity exercise of short (2-7 min) duration. If the muscle glycogen content is lowered by prolonged (1-1.5 h) exhausting cycle exercise, and is subsequently kept low for 3- 4 days by consumption of a diet deficient in carbohydrate (< 5% of total energy intake), there is a dramatic (∼10- 30%) reduction in exercise capacity during cycling sustainable for about 5 min. The same effect is observed if exercise is preceded by 3 - 4 days on a carbohydrate-restricted diet or by a 24 h total fast without prior depletion of the muscle glycogen. Consumption of a diet high in carbohydrate (70% of total energy intake from carbohydrate) for 3 - 4 days before exercise improves exercise capacity during high-intensity exercise, although this effect is less consistent. The blood lactate concentration is always lower at the point of fatigue after a diet low in carbohydrate and higher after a diet high in carbohydrate than after a normal diet. Even when the duration of the exercise task is kept constant, the blood lactate concentration is higher after exercise on a diet high in carbohydrate than on a diet low in carbohydrate. Consumption of a low-carbohydrate isoenergetic diet is achieved by an increased intake of protein and fat. A high-protein diet, particularly when combined with a low carbohydrate intake, results in metabolic acidosis, which ensues within 24 h and persists for at least 4 days. This appears to be the result of an increase in the circulating concentrations of strong organic acids, particularly free fatty acids and 3-hydroxybutyrate, together with an increase in the total plasma protein concentration. This acidosis, rather than any decrease in the muscle glycogen content, may be responsible for the reduced exercise capacity in highintensity exercise; this may be due to a reduced rate of efflux of lactate and hydrogen ions from the working muscles. Alternatively, the accumulation of acetyl groups in the carbohydrate-deprived state may reduce substrate flux through the pyruvate dehydrogenase complex, thus reducing aerobic energy supply and accelerating the onset of fatigue.

Fixed acid uptake by visceral organs during early endotoxemia.

Metabolic acidosis commonly occurs in patients with the sepsis syndrome (1). Acidosis correlates with increased mortality even in the absence of increased lactate levels and is a more reliable predictor of outcome (2). Although many studies have focused on the pathophysiology, clinical significance, and treatment of lactic acidosis (3–9), relatively few have investigated other sources of metabolic acidosis in sepsis. This is surprising because lactate ion accounts for less than 50% of the “unmeasured” anions present in patients with sepsis and metabolic acidosis (10). We have previously demonstrated that during normal physiologic conditions the liver takes up some of these other anions. However, following administration of endotoxin the liver switches to release of unexplained anions (11). This might account for the appearance of unexplained anions in the circulation of patients with sepsis (10) and liver disease (12) and may result in systemic acidosis if these anions accumulate in excess of strong cations. However, a non-anion gap metabolic acidosis occurs within several minutes of the administration of endotoxin to healthy animals (13). In addition, patients with sepsis frequently exhibit an acidosis which has a significant non-anion gap component (14). This acidosis cannot be explained by the release of anions by the liver. Accordingly, we reanalyzed the data from our original study (11) and included analysis of net acid flux in order to determine the sites of fixed acid production and clearance in acute endotoxemia.

Effect of lactic acid on postprandial plasma-glucose and -insulin responses in rats administered glucose solution

The effect of lactic acid or sodium lactate on the glycemic response was studied in rats. The lightly anesthetized, overnight-fasted rats were administered a glucose solution (200 g/L) with or without lactic acid or sodium lactate by a stomach tube to provide 250 mg of glucose per 100 g body weight. The addition of lactic acid (300 mmol/L) to a glucose solution reduced the initial rise in plasma glucose and insulin concentrations, slowed the rate of gastric emptying and brought a drop of pH in gastric contents. However, such effects were not recognized in the addition of sodium lactate (300 mmol/L as the lactate ion). Lactic acid stimulated cholecystokinin (CCK) and secretin release, but inhibited gastrin release. The study suggests that lactic acid is effective in modulating postprandial plasma glucose concentration, which is due to delayed gastric emptying accompanied by the increase of secretin and CCK release following the fall in intraduodenal pH.

Chloride binding in the stroma of cultured human corneas

In the ox cornea, more than half of the non-diffusible, matrix negative charge is derived from the binding of free chloride ions. Because the magnitude of the net matrix charge is the dominant factor which determines the degree of stromal swelling, we investigated whether this phenomenon, stromal chloride binding, also occurs in human corneal stroma. Intrastromal ion concentrations were measured with radio-isotopes when human (outdated Eye Bank) corneas or (fresh) bovine corneas, physically clamped to maintain a constant hydration, were incubated in buffered 154 mM NaCl. The intrastromal chloride ion concentration was compared to the normalized concentrations of trace quantities of radio-labelled acetate and lactate ions. For human corneas, the intrastromal chloride ion concentration was found to be significantly higher (P < 0.001, t-test) than the normalized concentrations of both acetate and lactate ([Cl]i = 142.5 +/- 0.9 mM, (n = 9); [acetate]i = 131.2 +/- 1.2 mM, (n = 8); [lactate]i = 131.9 +/- 1.5 mM, (n = 5); all values are mean +/- S.E.M.). The sodium ion concentration was elevated ([Na]i = 176.0 +/- 1.8 mM, (n = 9)). These results demonstrate that chloride binding occurs to a significant extent in cultured human corneal stroma and suggest that chloride binding may be evident in the native human cornea.

Increased [lactate] in working dog muscle reduces tension development independent of pH

The purpose of this work was to examine the effect of the lactate ion on the fatigue process in working muscle independent of muscle [H+]. L-(+)-lactate was infused, at a pH that did not change arterial pH, into the blood perfusing an isolated, in situ dog gastrocnemius (N = 5) working at a submaximal intensity (isometric contractions at 2 Hz) and compared with control (C) conditions without lactate infusion. Each muscle was stimulated to work for two 60-min periods (separated by 45 min rest), consisting of three 20-min time periods with either the high arterial lactate condition (high [La]) or C condition sequentially ordered within each 60-min work period. Blood flow and O2 delivery were held constant between the C and high [La] conditions. Arterial and venous blood measurements and muscle biopsies were taken (7 biopsies from each condition) during each condition. Lactate infusion significantly increased arterial [La] (C = 4.2 +/- 0.2 mM vs high [La] = 14.4 +/- 0.2; mean +/- SE) and muscle [La] (C = 8.1 +/- 0.8 mM w.w. vs high [La] = 12.0 +/- 1.4) while arterial and muscle pH were unchanged between conditions. Muscle tension development was significantly reduced (C = 94 +/- 2 N.100 g-1 vs high [La] = 80 +/- 3) during lactate infusion and muscle O2 uptake changed proportionally with tension. These findings support an effect of the lactate anion on tension development which is independent of pH.

Fixed Acid Uptake by Visceral Organs During Early Endotoxemia

Metabolic acidosis commonly occurs in patients with the sepsis syndrome (1). Acidosis correlates with increased mortality even in the absence of increased lactate levels and is a more reliable predictor of outcome (2). Although many studies have focused on the pathophysiology, clinical significance, and treatment of lactic acidosis (3–9), relatively few have investigated other sources of metabolic acidosis in sepsis. This is surprising because lactate ion accounts for less than 50% of the “unmeasured” anions present in patients with sepsis and metabolic acidosis (10). We have previously demonstrated that during normal physiologic conditions the liver takes up some of these other anions. However, following administration of endotoxin the liver switches to release of unexplained anions (11). This might account for the appearance of unexplained anions in the circulation of patients with sepsis (10) and liver disease (12) and may result in systemic acidosis if these anions accumulate in excess of strong cations. However, a non-anion gap metabolic acidosis occurs within several minutes of the administration of endotoxin to healthy animals (13). In addition, patients with sepsis frequently exhibit an acidosis which has a significant non-anion gap component (14). This acidosis cannot be explained by the release of anions by the liver. Accordingly, we reanalyzed the data from our original study (11) and included analysis of net acid flux in order to determine the sites of fixed acid production and clearance in acute endotoxemia.