Control of lactate oxidation in fish hearts by lactate oxidase activity

1985 ◽  
Vol 63 (3) ◽  
pp. 484-487 ◽  
Author(s):  
William R. Driedzic ◽  
John M. Stewart ◽  
Gwen McNairn

Isolated hearts of ocean pout (Macrozoarces americanus) and sea raven (Hemitripterus americanus) were perfused with media containing [14C]lactate or pyruvate and the rate of 14CO2 production was monitored. Increases in exogenous lactate concentration resulted in increases in the rate of lactate metabolism. Under comparable perfusion conditions the rate of decarboxylation of pyruvate was three- to four-fold higher than that of lactate. This finding suggests that lactate oxidation was being limited by lactate oxidase. LDH was purified and the Km values for lactate and pyruvate assessed under conditions of saturating cofactor concentration. Both hearts had a muscle type LDH on the basis of Km (pyruvate). Lactate oxidase from ocean pout and sea raven heart displayed Km values of 25 and 20 mM for lactate, respectively. The Km values were well above the presumptive intracellular level of lactate in the perfused hearts. Considered together, the perfusion and isolated enzyme studies show that the catabolism of exogenous lactate is limited by the reaction catalyzed by lactate oxidase.

1983 ◽  
Vol 61 (8) ◽  
pp. 1880-1883 ◽  
Author(s):  
William R. Driedzic ◽  
Donna L. Scott ◽  
Anthony P. Farrell

The relative contribution of aerobic and anaerobic metabolism to ATP production was assessed in sea raven (Hemitripterus americanus) hearts. The problem was approached by measuring the rates of oxygen consumption and lactate production by perfused isolated hearts performing mechanical work. In the experimental preparation aerobic metabolism could account for essentially all of the ATP synthesized; as such, the organization of metabolism in this fish heart appears similar to reptilian and mammalian hearts under conditions of adequate oxygen availability.


Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 208
Author(s):  
Hong Dinh Duong ◽  
Jong Il Rhee

In this study, ratiometric fluorescent glucose and lactate biosensors were developed using a ratiometric fluorescent oxygen-sensing membrane immobilized with glucose oxidase (GOD) or lactate oxidase (LOX). Herein, the ratiometric fluorescent oxygen-sensing membrane was fabricated with the ratio of two emission wavelengths of platinum meso-tetra (pentafluorophenyl) porphyrin (PtP) doped in polystyrene particles and coumarin 6 (C6) captured into silica particles. The operation mechanism of the sensing membranes was based on (i) the fluorescence quenching effect of the PtP dye by oxygen molecules, and (ii) the consumption of oxygen levels in the glucose or lactate oxidation reactions under the catalysis of GOD or LOX. The ratiometric fluorescent glucose-sensing membrane showed high sensitivity to glucose in the range of 0.1–2 mM, with a limit of detection (LOD) of 0.031 mM, whereas the ratiometric fluorescent lactate-sensing membrane showed the linear detection range of 0.1–0.8 mM, with an LOD of 0.06 mM. These sensing membranes also showed good selectivity, fast reversibility, and stability over long-term use. They were applied to detect glucose and lactate in artificial human serum, and they provided reliable measurement results.


2003 ◽  
Vol 284 (1) ◽  
pp. E193-E205 ◽  
Author(s):  
G. van Hall ◽  
M. Jensen-Urstad ◽  
H. Rosdahl ◽  
H.-C. Holmberg ◽  
B. Saltin ◽  
...  

To study the role of muscle mass and muscle activity on lactate and energy kinetics during exercise, whole body and limb lactate, glucose, and fatty acid fluxes were determined in six elite cross-country skiers during roller-skiing for 40 min with the diagonal stride (Continuous Arm + Leg) followed by 10 min of double poling and diagonal stride at 72–76% maximal O2 uptake. A high lactate appearance rate (Ra, 184 ± 17 μmol · kg−1 · min−1) but a low arterial lactate concentration (∼2.5 mmol/l) were observed during Continuous Arm + Leg despite a substantial net lactate release by the arm of ∼2.1 mmol/min, which was balanced by a similar net lactate uptake by the leg. Whole body and limb lactate oxidation during Continuous Arm + Leg was ∼45% at rest and ∼95% of disappearance rate and limb lactate uptake, respectively. Limb lactate kinetics changed multiple times when exercise mode was changed. Whole body glucose and glycerol turnover was unchanged during the different skiing modes; however, limb net glucose uptake changed severalfold. In conclusion, the arterial lactate concentration can be maintained at a relatively low level despite high lactate Ra during exercise with a large muscle mass because of the large capacity of active skeletal muscle to take up lactate, which is tightly correlated with lactate delivery. The limb lactate uptake during exercise is oxidized at rates far above resting oxygen consumption, implying that lactate uptake and subsequent oxidation are also dependent on an elevated metabolic rate. The relative contribution of whole body and limb lactate oxidation is between 20 and 30% of total carbohydrate oxidation at rest and during exercise under the various conditions. Skeletal muscle can change its limb net glucose uptake severalfold within minutes, causing a redistribution of the available glucose because whole body glucose turnover was unchanged.


1986 ◽  
Vol 60 (1) ◽  
pp. 232-241 ◽  
Author(s):  
R. S. Mazzeo ◽  
G. A. Brooks ◽  
D. A. Schoeller ◽  
T. F. Budinger

Lactate irreversible disposal (RiLa) and oxidation (RoxLa) rates were studied in six male subjects during rest (Re), easy exercise [EE, 140 min of cycling at 50% of maximum O2 consumption (VO2max)] and hard exercise (HE, 65 min at 75% VO2max). Twenty minutes into each condition, subjects received a Na+-L(+)-[1–13C]lactate intravenous bolus injection. Blood was sampled intermittently from the contralateral arm for metabolite levels, acid-base status, and enrichment of 13C in lactate. Expired air was monitored continuously for determination of respiratory parameters, and aliquots were collected for determination of 13C enrichment in CO2. Steady-rate values for O2 consumption (VO2) were 0.33 +/- 0.01, 2.11 +/- 0.03, and 3.10 +/- 0.03 l/min for Re, EE, and HE, respectively. Corresponding values of blood lactate levels were 0.84 +/- 0.01, 1.33 +/- 0.05, and 4.75 +/- 0.28 mM in the three conditions. Blood lactate disposal rates were significantly correlated to VO2 (r = 0.78), averaging 123.4 +/- 20.7, 245.5 +/- 40.3, and 316.2 +/- 53.7 mg X kg-1 X h-1 during Re, EE, and HE, respectively. Lactate oxidation rate was also linearly related to VO2 (r = 0.81), and the percentage of RiLa oxidized increased from 49.3% at rest to 87.0% during exercise. A curvilinear relationship was found between RiLa and blood lactate concentration. It was concluded that, in humans, 1) lactate disposal (turnover) rate is directly related to the metabolic rate, 2) oxidation is the major fate of lactate removal during exercise, and 3) blood lactate concentration is not an accurate indicator of lactate disposal and oxidation.


1976 ◽  
Vol 65 (1) ◽  
pp. 131-145 ◽  
Author(s):  
E. D. Stevens ◽  
A. M. Sutterlin

1. The ability of fish gills to transfer heat was measured by applying a heat pulse to blood in the ventral aorta and measuring it before and after passing through the gills of a teleost, Hemitripterus americanus. 2. 80–90% of heat contained in the blood is lost during passage through the gills. 3. The fraction of heat not lost during passage through the gills is due to direct transfer of heat between the afferent and efferent artery within the gill bar. 4. The major fraction of metabolic heat (70 - 90%) is lost through the body wall and fins of the sea raven in sea water at 5 degrees C; the remainder is lost through the gills.


1982 ◽  
Vol 242 (4) ◽  
pp. H485-H489 ◽  
Author(s):  
L. D. Segel ◽  
S. V. Rendig

Isolated working rat hearts were perfused with the perfluorochemical emulsion Fluosol-43 to determine whether it would support prolonged, stable cardiac function. The perfluorochemical emulsion provides a controlled perfusate composition, relatively high oxygen capacity, and a colloid osmotic pressure that is similar to that of plasma. Electrically paced (325 beats/min) hearts were perfused for 6 h at 35 degrees C in a recirculating system. Hemodynamic and mechanical function of seven Krebs-Henseleit-perfused hearts declined significantly more than that of seven Fluosol-43-perfused hearts over the 6-h period. The percent of initial function remaining at 6 h for Krebs-Henseleit-perfused vs. Fluosol-43-perfused hearts was 70.3 +/- 5.0 vs. 95.4 +/- 1.1% (P less than 0.001) for left ventricular peak systolic pressure; 55.6 +/- 7.7 vs. 97.5 +/- 2.4% (P less than 0.001) for dP/dtmax; 27.1 +/- 7.2 vs. 60.6 +/- 5.2% (P less than 0.005) for cardiac output; 11.0 +/- 6.3 vs. 67.2 +/- 3.4% (P less than 0.001) for external work efficiency; and 17.4 +/- 8.4 vs. 67.7 +/- 4.5% (P less than 0.001) for stroke rhythmic than Fluosol-43-perfused hearts during the last 3 h of perfusion. Perfusate lactate concentration was 4.5-fold higher in the Krebs-Henseleit perfusate than in the Fluosol-43 perfusate at 6 h, reflecting greater anaerobic metabolism in the Krebs-Henseleit-perfused hearts. Thus isolated rat hearts perfused with Fluosol-43 have greater maintenance of hemodynamic and mechanical function over a longer time period than hearts perfused with Krebs-Henseleit bicarbonate buffer.


Perfusion ◽  
2002 ◽  
Vol 17 (3) ◽  
pp. 167-173 ◽  
Author(s):  
Y M Ganushchak ◽  
J G Maessen ◽  
D S de Jong

Background: Patients undergoing cardiac surgery with the use of cardiopulmonary bypass (CPB) are often thought to have tissue hypoxia and intraoperative oxygen debt accumulation despite the lack of sufficient data to support this assumption. Methods and results: Oxygen uptake and related parameters, including the plasma lactate and pyruvate concentrations, were studied during the peri-operative period in a group of 15 consecutive patients who underwent coronary artery bypass graft surgery. The actual oxygen uptake (VO2) and delivery (DO2) were compared with the individual expected (computed) oxygen transport values. The mean values of DO2 and VO2were in the range of the expected values. Our results demonstrate a leading role for body temperature in perioperative changes of oxygen consumption rate ( r2 = 0.65, p< 0.001). Plasma lactate and pyruvate did not exceed the physiological range in any patient. However, with initiation of CPB, the lactate to pyruvate (LA/PVA) ratio increased (from 9.87± 2.43 at T1 to 12.08± 1.51 at T2, p< 0.05). The mean value of the LA/PVA ratio was elevated during surgery. Later, upon lowering of the plasma lactate concentration in the postoperative period, the LA/PVA ratio decreased to normal values. Without any other evidence of hypoxia, this increase in the LA/PVA ratio could be explained by washout of lactate from previously hypoperfused tissues and intraoperative decrease of lactate clearance. Conclusion: Systemic oxygenation was not impaired during CPB, or during 18 h after surgery in the studied group of patients.


1986 ◽  
Vol 251 (6) ◽  
pp. R1144-R1150 ◽  
Author(s):  
J. R. Bailey ◽  
W. R. Driedzic

Myoglobin, an intracellular O2-binding protein, plays a protective role in maintaining performance of isolated fish hearts under hypoxic conditions. This study was designed to test the hypothesis that the protein contributes to O2 consumption under conditions of increased O2 demand or hypoxia. Isolated myoglobin-rich sea raven (Hemitripterus americanus) hearts and myoglobin-poor ocean pout (Macrozoarces americanus) hearts were perfused under conditions of changing partial pressure of O2 (PO2) and afterload. Sea raven hearts maintained O2 consumption and cardiac performance at low PO2 and high afterload, whereas ocean pout hearts did not. In other cases sea raven and ocean pout hearts were treated with hydroxylamine, which renders myoglobin incapable of binding O2, and subjected to changing PO2 and afterload. Sea raven hearts could not maintain O2 consumption and cardiac performance, whereas hydroxylamine treatment had no effect on O2 consumption in ocean pout hearts under these conditions. These data provide the first evidence to support the concept that myoglobin plays a role in O2 consumption of hearts.


1975 ◽  
Vol 228 (6) ◽  
pp. 1656-1662 ◽  
Author(s):  
AC Beardsley ◽  
AM Lefer

Isolated hearts form chronically adrenalectomized cats were perfused with Krebs-Henseleit buffer plus either glucose (10mM) or palmitate (0.4 mM) under various conditions of constant pressure and constant flow. Glucose uptake in adrenalectomizedhearts was not diminished from control values under conditions of constant pressure, constant flow, anoxia, or insulin stimulation. Palmatic acid uptake and oxygen consumption were significantly reduced (P less than 0.02) in adrenalectomized hearts. This diminished fatty acid utilization was also reflected in a significantly lower CO'2 production and incorporation of the palmitate into myocardial triglycerides. The decreased fatty acid uptake by adrenalectomized cat hearts may represent aserious defect in myocardial metabolism since lipids are the major energy substrate forthe heart. Whether the defect occurs in fatty acid transport or activation cannot beelucidated by this study. However, it is unlikely that this defect has a major contributory effect on the dysfunction of adrenalectomized hearts since the myocardium iscabable of using other energy substrates readily.


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