Modeling cerebral arteriovenous lactate kinetics after intravenous lactate infusion in the rat.

Venous-arterial lactate differences across the brain during lactate infusion in rats were studied, and the fate of lactate was described with a mathematical model that includes both cerebral and extracerebral kinetics. Ultrafiltration was used to sample continuously and simultaneously arterial and venous blood. Subsequent application of flow injection analysis and biosensors allowed the measurement of glucose and lactate concentrations every minute. Because of the high temporal resolution, arteriovenous lactate kinetics could be modeled in individual experiments. The existence of both a cerebral lactate sink and a lactate exchangeable compartment, representing approximately 24% of brain volume, was thus modeled.

The potential of biosensor technology in clinical monitoring and experimental research.

Glucose or lactate biosensors are very useful for monitoring metabolism. Continuous monitoring of glucose is for example very important in diabetic patients. The measurement of lactate, a marker for oxygen deficiency, is used in the intensive care unit to monitor the patients' condition. In our laboratory we have developed two types of on-line biosensors to measure in vivo glucose and lactate: a sandwich-type biosensor and, very recently, a miniaturized flow-through biosensor. These biosensors are not placed in the body itself, but are connected to implanted microdialysis or ultrafiltration probes. Both types of biosensors are based on the oxidation of substrate using glucose oxidase or lactate oxidase and electrochemical detection. In the sandwich-type sensor, the enzymes are physically immobilized between two cellulose nitrate filters, and operate with ferrocene as a mediator. In the miniaturized biosensor, with an internal volume of 10-20 nanolitres, the enzymes are immobilized on the electrode via in situ encapsulation in poly(1,3-phenylenediamine). In this review we shall explain the working of these biosensors, and describe their application in clinical monitoring and experimental research.

Quantitative on-line monitoring of cellular glucose and lactate metabolism in vitro with slow perfusion.

An on-line in vitro perfusion technique is described that allows the continuous quantification of cellular glucose metabolism in vitro. Using biosensor technology, we measure glucose and lactate metabolism at a minute-to-minute time resolution for periods up to several days. The application of our perfusion-detection technique for in vitro monitoring is demonstrated in a wide variety of cells, including primary neuronal and astroglia cultures, yeast cells, and human lymphocytes. The method shows that variations in oxygen delivery or exposure to a noncompetitive pseudosubstrate (here 2-deoxyglucose) affects normal glucose metabolism. The innovative advantage of the present system is that, in contrast to other devices including a recently described system, metabolism per cell can be quantified. The potential of in vitro on-line monitoring is discussed for application in studying normal and abnormal metabolism, toxic and nontoxic drug effects, and human tissue biopsies.

Utilization of in vivo ultrafiltration in biomedical research and clinical applications.

Ultrafiltration (UF) is a filtrate selection method with a wide range of biomedical and clinical applications, including detoxification of blood in hemodialysis and peritoneal dialysis. New is, however, the use of UF as a convenient in vivo sampling method that, for example, has been used in diabetics. Ultrafiltration avoids complicated and time-consuming recovery calculations that are necessary when using in vivo microdialysis, as recoveries of low molecular weight molecules are near 100%. The subcutaneously or intravenously placed UF probes have been studied for off-line sample analysis and for continuous on-line monitoring, in a wide variety of species, including dogs, rats, pigs and humans. This review discusses the potential of in vivo UF as a continuous tissue sampling technique in clinical research areas, and in several major biomedical applications including glucose and lactate monitoring and drug kinetic studies.

Evidence for a lactate pool in the rat brain that is not used as an energy supply under normoglycemic conditions.

Lactate derived from glucose can serve as an energy source in the brain. However, it is not certain how much lactate, directly taken from the blood circulation, may replace glucose as an energy source. This study aimed to estimate the uptake, release, and utilization of lactate entering the brain from the blood circulation. The change in cerebral venous-arterial glucose and lactate differences after lactate infusions in the anesthetized rat were measured. Ultrafiltration probes were placed in the aorta and in the jugular vein, and connected to a flow injection analysis system with biosensors for glucose and lactate. Measurements were taken every minute. Lactate efflux was observed at baseline, whereas an influx of lactate was seen during lactate infusion. Immediately after the infusion there was a net efflux of lactate from the brain. The results suggest that the majority of lactate moving into the brain is not used as an energy substrate, and that lactate does not replace glucose as an energy source. Instead, the authors propose the concept of a lactate pool in the brain that can be filled and emptied in accordance with the blood lactate concentration, but which is not used as an energy supply for cerebral metabolism.

Quantitative on-line monitoring of hippocampus glucose and lactate metabolism in organotypic cultures using biosensor technology.

Quantitative glucose and lactate metabolism was assessed in continuously perfused organotypic hippocampal slices under control conditions and during exposure to glutamate and drugs that interfere with aerobic and anaerobic metabolism. On-line detection was possible with a system based on slow perfusion rates, a half-open (medium/air interface) tissue chamber and a flow injection analytic system equipped with biosensors for glucose and lactate. Under basal conditions about 50% of consumed glucose was converted to lactate in hippocampal slice cultures. Using medium containing lactate (5 mm) instead of glucose (5 mm) significant lactate uptake was observed, but this uptake was less than the net uptake of lactate equivalents in glucose-containing medium. Glucose deprivation experiments suggested lactate efflux from glycogen stores. The effects of drugs compromising or stimulating energy metabolism, i.e. 2-deoxyglucose, 3-nitropropionic acid, alpha-cyano-4-hydroxycinnamate, l-glutamate, d-asparate, ouabain and monensin, were tested in this flow system. The data show that maintaining Na+ and K+ gradients consumed much of the energy but do not support the hypothesis that l-glutamate stimulates glycolysis in hippocampal slice cultures.