Presence of lactate dehydrogenase and lactate racemase in Megasphaera elsdenii grown on glucose or lactate.

Activity of D-lactate dehydrogenase (D-LDH) was shown not only in cell extracts from Megasphaera elsdenii grown on DL-lactate, but also in cell extracts from glucose-grown cells, although glucose-grown cells contained approximately half as much D-LDH as DL-lactate-grown cells. This indicates that the D-LDH of M. elsdenii is a constitutive enzyme. However, lactate racemase (LR) activity was present in DL-lactate-grown cells, but was not detected in glucose-grown cells, suggesting that LR is induced by lactate. Acetate, propionate, and butyrate were produced similarly from both D- and L-lactate, indicating that LR can be induced by both D- and L-lactate. These results suggest that the primary reason for the inability of M. elsdenii to produce propionate from glucose is that cells fermenting glucose do not synthesize LR, which is induced by lactate.     

Characterization of rabbit lactate dehydrogenase-M and lactate dehydrogenase-H cDNAs. Control of lactate dehydrogenase expression in rabbit muscle

Two cDNA clones were isolated, one corresponding to the mRNA coding for lactate dehydrogenase-M (LDH-M), the other to the mRNA coding for lactate dehydrogenase-H (LDH-H). The cDNA inserts consist of the entire open reading frame for LDH-M and a partial sequence, from amino acid 117 to 332, for LDH-H. Using these two clones as probes we demonstrate that: (a) the abundance of mRNA is muscle-type dependent; (b) the ratio M/H subunit for protein and mRNA is well related in the muscles studied; and (c) the M + H mRNA level is not relative to the total LDH activity.     

Autophosphorylation of lactate dehydrogenase

Incubation of rabbit muscle lactate dehydrogenase in the presence of Mg[alpha-32p]ATP results in the incorporation of the label into the protein. The autophosphorylation reaction is strongly pH-dependent. The maximal phosphorylation is observed at pH 6.8 with 3-4 moles of phosphate bound per mole of tetrameric enzyme. The enzyme-phosphate complex is readily hydrolyzed by hydroxylamine.     

Transpulmonary lactate shuttle

The shuttling of intermediary metabolites such as lactate through the vasculature contributes to the dynamic energy and biosynthetic needs of tissues. Tracer kinetic studies offer a powerful tool to measure the metabolism of substrates like lactate that are simultaneously taken up from and released into the circulation by organs, but in each circulatory passage, the entire cardiac output traverses the pulmonary parenchyma. To determine whether transpulmonary lactate shuttling affects whole-body lactate kinetics in vivo, we examined the effects of a lactate load (via lactate clamp, LC) and epinephrine (Epi) stimulation on transpulmonary lactate kinetics in an anesthetized rat model using a primed-continuous infusion of [U-(13)C]lactate. Under all conditions studied, control 1.2 (SD 0.7) (Con), LC 1.9 (SD 2.5), and Epi 1.9 (SD 3.5) mg/min net transpulmonary lactate uptake occurred. Compared with Con, a lactate load via LC significantly increased mixed central venous ([v]) [1.9 mM (SD 0.5) vs. 4.7 (SD 0.4)] and arterial ([a]) [1.6 mM (SD 0.4) vs. 4.1 (SD 0.6)] lactate concentrations (P < 0.05). Transpulmonary lactate gradient ([v] - [a]) was highest during the lactate clamp condition [0.6 mM (SD 0.7)] and lowest during Epi [0.2 mM (SD 0.5)] stimulation (P < 0.05). Tracer measured lactate fractional extractions were similar for control, 16.6% (SD 15.3), and lactate clamp, 8.2% (SD 15.3) conditions, but negative during Epi stimulation, -25.3% (SD 45.5) when there occurred a transpulmonary production, the conversion of mixed central venous pyruvate to arterial lactate. Further, isotopic equilibration between L and P occurred following tracer lactate infusion, but depending on compartment (v or a) and physiological stimulus, [L]/[P] concentration and isotopic enrichment ratios ranged widely. We conclude that pulmonary arterial-vein concentration difference measurements across the lungs provide an incomplete, and perhaps misleading picture of parenchymal lactate metabolism, especially during epinephrine stimulation.     

Needle-type lactate biosensor

A needle-type lactate biosensor has been developed for continuous intravascular lactate monitoring. The sensor employs poly(1,3-phenylenediamine) as the inner layer on the platinum electrode in order to eliminate the interference from oxidizable physiological substances. Cross-linking with glutaraldehyde was used for enzyme immobilization. Dithiothreitol was used as the stabilizer of lactate oxidase. PVC (polyvinyl chloride) was chosen as the external diffusion control membrane. Sensor performance was evaluated in vitro and the sensor shows a sensitivity of 10-15 nA/mM, and a linear range from 1 mM to at least 15 mM lactate. Evaluation of the sensor response in blood plasma showed similar sensitivity and linear range as indicated by the calibration curves obtained in buffer solution. The sensor has a short response time of approximately 1 minute. The sensors were operated continuously for 7 days in phosphate buffer containing solution with a concentration at the physiological lactate level. No significant change in sensor sensitivity and its linear range has been observed. Sensors show a minimum change in its performance when stored in buffer at 4 degrees C for at least 9 months.     

Measurements of lactate concentration using lactate oxidase and an electrochemical oxygen sensor

Lactate oxidase was used in combination with an electrochemical dissolved oxygen sensor to measure L-lactate concentration in the physiological saline solution. The rate of oxygen consumption was found to have an excellent linear relationship with the lactate concentration on a log-log scale in the lactate concentration range of 0.3-25 mM. This detection does not require additional reagents and can be developed into a simple method of L-lactate determination. The effects of the temperature and pH of the solution on the reaction rate were investigated and discussed.     

Effects of a 30-km race upon salivary lactate correlation with blood lactate

Blood lactate has been used to determine the aerobic capacity and long distance performance. Recently, a new methodology has been suggested to supplant the invasive blood lactate techniques. Salivary lactate has received attention because it shows high correlation to blood lactate in progressive overload test. We evaluated the correlation between salivary and blood lactate during a long distance run and assessed possible changes in salivary lactate concentration. Fifteen expert marathon racers ran 30 km as fast as possible. Saliva and 25 muL of blood were collected at rest and at each 6 km for lactate determination. Blood lactate concentration increased in the 6th km and then remained constant until the end of the race. Salivary lactate increased after 18 km in relation to basal. We found high correlations between blood and saliva absolute lactate (r=0.772, p<0.05) and the blood lactate relative concentration corrected by protein (r=0.718, p<0.05). The highest correlation found between absolute and relative salivary lactate was r=0.994 (p<0.001). Our results show that it is possible to use salivary lactate with absolute values or relative protein concentration. In addition, salivary lactate showed a high correlation with blood lactate in endurance events.     

Metabolic regulation by lactate

For more than a century, the metabolic role of lactate has intrigued physiologists and biochemists. Yet, for the first half of the last century lactate had been designated as a waste product, and assigned no additional significance besides its controversial role in muscle fatigue. The decline of the lactate hypothesis for the onset of muscle fatigue and the defining of some modulatory properties attributed to lactate have increased the interest on this molecule. The present critical review aimed at evaluating some recent publications concerned with unveiling the regulatory actions of lactate in cellular function. Lactate has been described to modulate enzymes catalytic properties to affect hormonal release and responsiveness, and to control body homeostasis. Moreover, these properties are directly related to the genesis and the sustainability of pathological conditions, such as diabetes and cancer. In the end, we concluded that lactate should not be regarded as simply an anaerobic metabolite, but should be considered as a regulatory molecule that modulates the integration of metabolism.     

The movement of pyruvate, lactate and lactate dehydrogenase into rabbit oviductal fluid.

Pyruvate, lactate and lactate dehydrogenase appeared linearly in 2 ml 0.9% NaCl recirculated through the rabbit oviduct for 4 h in vivo. In oviducts from rabbits injected 3 days previously with 100 i.u. hCG, the rate of appearance of all three constituents was considerably reduced. It is considered unlikely that the lactate dehydrogenase secreted brings about the interconversion of pyruvate and lactate in the oviduct lumen.     

Protein fluorescence of lactate dehydrogenase

1. There is a non-linear decrease in the protein fluorescence (F) of lactate dehydrogenase with the increase in the fraction (alpha) of the coenzyme-binding sites occupied with NADH. 2. By a curve-fitting procedure it is shown that the fluorescence intensity can be represented by the equation F=[1-alpha(1-x)](n) where n is the number of identical and indistinguishable coenzyme-binding sites per protein molecule and x=F(s) (1/n) (F(s) is the protein fluorescence at alpha=1). This equation implies that the relative protein fluorescence of molecules bearing j ligands form the geometric series x(j). 3. Non-linear quenching of protein fluorescence for this enzyme is probably due to radiationless transfer of energy from the protein molecule to the bound NADH and should also be observed when other potential acceptors of protein fluorescence are bound at unique sites. 4. The intercept with F(s) of an initial tangent to a curve of protein fluorescence against alpha will be at a value of alpha equal to (K(d)+[E(0)]). (1-x(n))/n.(1-x) and not at a value equal to the sum of the dissociation constant (K(d)) and the concentration of identical ligand-binding sites ([E(0)]). 5. A use of non-linear protein fluorescence quenching to investigate the state of aggregation of a protein is discussed.     

Hepatic lactate uptake versus leg lactate output during exercise in humans.

The exponential rise in blood lactate with exercise intensity may be influenced by hepatic lactate uptake. We compared muscle-derived lactate to the hepatic elimination during 2 h prolonged cycling (62 +/- 4% of maximal O(2) uptake, (.)Vo(2max)) followed by incremental exercise in seven healthy men. Hepatic blood flow was assessed by indocyanine green dye elimination and leg blood flow by thermodilution. During prolonged exercise, the hepatic glucose output was lower than the leg glucose uptake (3.8 +/- 0.5 vs. 6.5 +/- 0.6 mmol/min; mean +/- SE) and at an arterial lactate of 2.0 +/- 0.2 mM, the leg lactate output of 3.0 +/- 1.8 mmol/min was about fourfold higher than the hepatic lactate uptake (0.7 +/- 0.3 mmol/min). During incremental exercise, the hepatic glucose output was about one-third of the leg glucose uptake (2.0 +/- 0.4 vs. 6.2 +/- 1.3 mmol/min) and the arterial lactate reached 6.0 +/- 1.1 mM because the leg lactate output of 8.9 +/- 2.7 mmol/min was markedly higher than the lactate taken up by the liver (1.1 +/- 0.6 mmol/min). Compared with prolonged exercise, the hepatic lactate uptake increased during incremental exercise, but the relative hepatic lactate uptake decreased to about one-tenth of the lactate released by the legs. This drop in relative hepatic lactate extraction may contribute to the increase in arterial lactate during intense exercise.     

Maximal lactate steady-state prediction through quadratic modeling of selected stages of the lactate minimum test

In this study, we compared the maximal lactate steady state (MLSS) with lactate minimal (LM) intensities determined visually and through a quadratic polynomial function of selected stages of LM test. Eleven male recreational cyclists (27.7 +/- 4.5 years, 175.7 +/- 5.6 cm, 69.5 +/- 10.8 kg, and 12.0 +/- 5.5% body fat) performed one LM test under previous induction of hyperlactaemia with an initial intensity of 75 W with 30-W increments every 3 minutes with blood lactate concentration (HLa) and rating of perceived exertion (RPE) measurements. The LM intensity was determined visually (VLM) and by modeling the lactate response through polynomial function by using: 1) all stages (LMP); 2) the first stage, the stage corresponding to RPE-13 and the last stage/exhaustion (LMP3max); 3) the three lowest lactate concentration stages (LMP3adj); and 4) the initial, RPE-13, and RPE-16 stages (LMP3sub). The MLSS was determined as the highest intensity at a variation not greater than 0.05 mmol.l.min of HLa during the last 20 minutes of a 30-minute exercise session. The MLSS (204.0 +/- 16.0 W), VLM (198.6 +/- 15.2 W), LMP3adj (190.4 +/- 12.9 W), and LMP3sub (192.1 +/- 27.2 W) were not different, well correlated, and in agreement to each other. In conclusion, the polynomial modeling of HLa response to three submaximal stages produced exercise intensities that did not differ from MLSS.     

Reliability of urine lactate as a novel biomarker of lactate production capacity in maximal swimming

Context: Postexercise urine lactate may be a novel biomarker of lactate production capacity during exercise.

Objective: To evaluate the reliability and utility of the urine lactate concentration after maximal swimming trials between different training protocols (6?×?50?m and 3?×?100?m) and training states (active and nonactive swimmers).

Materials and methods: Lactate and creatinine were determined by spectrophotometry in blood and urine.

Results: Blood and urine lactate concentrations were correlated in-between training protocols and in participants of different training states. The reliability of the urine lactate concentration was moderate for one of the training protocols and good or moderate for the two training states. Additionally, it was lower than that of the blood lactate concentration, and did not improve after normalizing to the urine creatinine concentration.

Discussion and conclusion: Although promising as a biomarker of lactate production capacity, urine lactate requires further research to improve its reliability.