The involvement of lactate and calcium as mediators of the electrical and mechanical responses of the myocardium to conditions of simulated ischaemia.

1. Rat isolated and superfused atria were exposed to a lactate-containing solution simulating the composition of extracellular fluid during myocardial ischaemia (SI). 2. Atria subjected to SI showed a decreased adenosine 5'-triphosphate (ATP) content, a rise in diastolic tension, a diminished conduction velocity of action potentials and shortened refractory periods. All these changes were less pronounced during lactate-free SI. 3. Atria preloaded with calcium displayed exaggerated responses measured electrically and mechanically during exposure to SI, whereas atria previously depleted of calcium displayed diminished electrical and mechanical responses to SI. Neither calcium loading nor calcium depletion modified the SI-induced depletion of the atrial stores of ATP. 4. Sulphinpyrazone protected atria against all aspects of the response to SI, but failed to protect the muscle under conditions of lactate-free SI. It is concluded that during SI, sulphinpyrazone protects against a lactate-mediated inhibition of the glycolytic synthesis of ATP. 5. Flufenamate exaggerated all responses of the atria to SI. These deleterious actions were still observed during lactate-free SI. It is concluded that flufenamate inhibits the synthesis of ATP in the mitochondria.

Possible mechanisms of the anaerobic threshold. A review.

The anaerobic threshold consists of a lactate threshold and a ventilatory threshold. In some conditions there may actually be 2 ventilatory thresholds. Much of the work detailing the lactate threshold is strongly based on blood lactate concentration. Since, in most cases, blood lactate concentration does not reflect production in active skeletal muscle, inferences about the metabolic state of contracting muscle will not be valid based only on blood lactate concentration measurements. Numerous possible mechanisms may be postulated as generating a lactate threshold. However, it is very difficult to design a study to influence only one variable. One may ask, does reducing F1O2 cause an earlier occurrence of a lactate threshold during progressive exercise by reducing oxygen availability at the mitochondria? By stimulating catecholamine production? By shifting more blood flow away from tissues which remove lactate from the blood? Or by some other mechanism? Processes considered essential to the generation of a lactate threshold include: (a) substrate utilisation in which the ability of contracting muscle cells to oxidise fats reaches maximal power at lactate threshold; and (b) catecholaminergic stimulation, for without the presence of catecholamines it appears a lactate threshold cannot be generated. Other mechanisms discussed which probably enhance the lactate threshold, but are not considered essential initiators are: (a) oxygen limitation; (b) motor unit recruitment order; (c) lactate removal; (d) muscle temperature receptors; (e) metabolic stimulation; and (f) a threshold of lactate efflux. Some mechanisms reviewed which may induce or contribute to a ventilatory threshold are the effects of: (a) the carotid bodies; (b) respiratory mechanics; (c) temperature; and (d) skeletal muscle receptors. It is not yet possible to determine the hierarchy of effects essential for generating a ventilatory threshold. This may indicate that the central nervous system integrates a broad range of input signals in order to generate a non-linear increase in ventilation. Evidence indicates that the occurrence of the lactate threshold and the ventilatory threshold may be dissociated; sometimes the occurrence of the lactate threshold significantly precedes the ventilatory threshold and at other times the ventilatory threshold significantly precedes the lactate threshold. It is concluded that the 2 thresholds are not subserved by the same mechanism.

Regulation of T-cell functions by L-lactate.

Lactate is a product of glycolytically active macrophages. After stimulation with concanavalin A accessory cell-depleted splenic T-cell populations were found to produce only minute amounts of T-cell growth factor (TCGF); but substantial amounts of TCGF were produced if the cultures were supplemented either with splenic adherent cells or with lactate but not with interleukin-1 (IL-1). IL-1 was capable, however, of supporting TCGF production by the thymoma subline EL4-6.1. TCGF production in cultures of accessory cell-depleted splenic T-cell populations was demonstrable with 10(-3) M L-lactate, and optimal responses (plateau level) were obtained with 4-6 X 10(-2) M L-lactate. Cultures of macrophages were found to accumulate up to 5 X 10(-2) M lactate. Our experiments indicate, therefore, that lactate serves as a regulatory signal by which macrophage-like accessory cells enhance helper-T-cell functions. Lactate is apparently not the only mediator of accessory cell function since plateau levels of TCGF production were markedly lower with lactate than with splenic accessory cells; but L-lactate was found also to determine the magnitude of T-cell-mediated immune responses in vivo and in cultures of unfractionated lymphocyte populations. The production of interferon in accessory cell-depleted and concanavalin A-treated T-cell cultures, however, was not significantly affected by lactate. Concanavalin A-stimulated splenic T-cell populations were found to consume glucose rapidly and to release lactate into the supernatant. This indicates that the cells contain more lactate and pyruvate than they can utilize by their respiratory metabolism. The administration of external lactate or pyruvate was found to inhibit the utilization of glucose by the mitogenically stimulated T cells.

Effects of dichloroacetate in spinal stroke in the rabbit.

High levels of brain lactate may contribute to cellular death and dysfunction in acute cerebral ischemia. Although sodium dichloroacetate (DCA) has been shown to lower brain lactate in incomplete cerebral ischemia, functional outcome has not been assessed with DCA. We examined the effects of DCA treatment on functional neurologic outcome using a previously developed model for "spinal stroke" in the rabbit. Thirty male New Zealand white rabbits weighing 1.3-2.8 kg were studied. After anesthesia with 15-40 mg/kg pentobarbital IV, a laparotomy was performed and the aorta exposed. A metal clamp was placed on the aorta just distal to the left renal artery for 20 minutes and then removed. The abdominal wound was closed in two layers. Animals then received either 2cc normal saline (n = 15) or 300 mg/kg DCA in 2cc normal saline (n = 15) over 10 minutes. The animals were returned to their cages when awake and were examined at 24 hours, 48 hours, and 72 hours for neurologic assessment. The exams were performed by a blinded examiner who was unaware of the treatment given. A three point ambulatory score (0 = can't walk, 1 = walk but not hop, 2 = hopping) and a two point activity score (0 = inactive, 1 = active) were used. At 24 hours, 67% of the DCA-treated animals were actively moving about compared to only 27% of the controls (P = 0.03; Fisher Exact Test). Ten of fifteen control animals were unable to walk, while only five of fifteen DCA-treated animals were unable to walk (P = 0.07). Sixty percent of the DCA animals were able to hop compared to 27% of controls (P = 0.06). These results suggest that DCA can reduce morbidity from spinal cord ischemia in the rabbit.

Can lactate be used as a fuel by wounded tissue?

The role of lactate in the metabolism of the healing wound is poorly understood. The purpose of the present studies was to determine if despite a net lactate production, wounded (Wx) tissue could metabolize lactate and use it as an oxidative fuel. The extensor digitorum longus muscles (EDL) of weanling, male, Fischer rats were injured with lambda-carrageenan or injured thermally, and 5 and 3 days later, respectively, were incubated in a standard incubate that contained varying amounts of lactate (0 to 6 mmol/L added). Lactate uptake and oxidation, occurred in lambda-carrageenan Wx EDL, thermally injured EDL and non-Wx EDL in a dose-dependent manner. At lactate concentrations of less than 3 mmol/L in the incubate, there was net lactate production, but at lactate concentrations of 6 mmol/L there was no net lactate production by both Wx and non-Wx EDL. The increase in lactate oxidation was not associated with an alteration in the tissue content of adenosine triphosphate or creatine phosphate. It was associated with a reduction in glucose oxidation in Wx and non-Wx EDL and by a decrease in glucose uptake by Wx EDL. These data suggest that lactate may be used as an oxidative fuel by wounded tissue and in this regard may substitute for glucose.

Contact lens-induced corneal endothelial polymegathism: functional significance and possible mechanisms.

The corneal endothelium is principally responsible for maintenance of corneal deturgescence. Therefore, compromise of corneal endothelial functional integrity can result in corneal swelling and opacification. Contact lenses constitute a potential insult to the cornea because their wear reduces the oxygen available to that tissue. It has been reported that contact lens wear induces transient as well as permanent morphologic changes in the corneal endothelium. One of the permanent changes reported is referred to as polymegathism, which is a variation in cell size within the endothelial monolayer. Several investigators have suggested that polymegathism reflects a compromised endothelial functional status. Mechanisms proposed to explain contact lens-induced polymegathism include lactate accumulation, changes in pH, and elevation in CO2 content. We discuss these possibilities as well as speculate that these polymegathous shape changes may be a result of decreased endothelial ATP (adenosine triphosphate) levels and disturbed calcium homeostasis due to corneal endothelial hypoxia.

Saccharomyces cerevisiae contains two functional citrate synthase genes.

The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.

[Psychopharmacology of panic attacks]. / Psychopharmacologie des attaques de panique.

A new type of anxiety state has recently been identified: the panic attack. This condition is qualitatively distinct from other anxiety disturbances and its treatment is different. Sodium lactate seems to play an essential role in the triggering of panic attacks through the intermediary of catabolic, physicochemical mechanisms or by alteration of adrenergic neurotransmission. Anxiety may be autonomised into two sub-groups, based on the effects of psychotropic drugs: phobic and anxiety states. The presence or absence of panic attacks is essential as it has diagnostic and therapeutic implications: M.A.D.I. or tricyclic antidepressors have a specific action on panic attacks but are ineffective on anticipatory, chronic, phobic or other forms of anxiety. Which are improved by benzodiazepines which; themselves, have very little effect on panic attacks.

Gluconeogenesis in tumor-influenced hepatocytes.

The growth of a tumor leads to alterations in host carbohydrate metabolism. In this study we examined gluconeogenic capacity and amino acid transport in tumor-influenced and control rat hepatocytes. Serum glucose level decreased with increasing tumor burden and a significant correlation (r = -0.80) was observed. Hepatic glycogen content was similar in both groups after an overnight fast. Endogenous glucose production was 27% higher in tumor-influenced hepatocytes. The presence of 10mM of alanine led to 72% stimulation of gluconeogenesis in tumor-influenced hepatocytes as compared to 48% stimulation in control hepatocytes. The same trends were present when lactate was used as a substrate. Alanine transport into the cells was increased in tumor-influenced hepatocytes by 55% +/- 5% at a physiologic level of substrate. In conclusion, gluconeogenesis from alanine and lactate is significantly increased in tumor-influenced hepatocytes despite decreased serum glucose levels. This is associated with increased gluconeogenic capacity and accelerated alanine transport.

The stimulation of hepatic gluconeogenesis by acetoacetate precursors. A role for the monocarboxylate translocator.

The regulation of the gluconeogenic pathway from the 3-carbon precursors pyruvate, lactate, and alanine was investigated in the isolated perfused rat liver. Using pyruvate (less than 1 mM), lactate, or alanine as the gluconeogenic precursor, infusion of the acetoacetate precursors oleate, acetate, or beta-hydroxybutyrate stimulated the rate of glucose production and, in the case of pyruvate (less than 1 mM), the rate of pyruvate decarboxylation. alpha-Cyanocinnamate, an inhibitor of the monocarboxylate transporter, prevented the stimulation of pyruvate decarboxylation and glucose production due to acetate infusion. With lactate as the gluconeogenic precursor, acetate infusion in the presence of L-carnitine stimulated the rate of gluconeogenesis (100%) and ketogenesis (60%) without altering the tissue acetyl-CoA level usually considered a requisite for the stimulation of gluconeogenesis by fatty acids. Hence, our studies suggest that gluconeogenesis from pyruvate or other substrates which are converted to pyruvate prior to glucose synthesis may be limited or controlled by the rate of entry of pyruvate into the mitochondrial compartment on the monocarboxylate translocator.

Relation between membrane potential and lactate in gastrocnemius and soleus muscle of the cat during tourniquet ischemia and postischemic reflow.

The relation between the lactate content and the membrane potential was investigated in the gastrocnemius and soleus muscle of the cat during and after a 4h period of ischemia. The skeletal muscle content of ATP and glucose was also measured. No change occurred in the ATP content of the gastrocnemius muscle during ischemia and increased above initial values after reflow in both muscles. The lactate content increased, and the membrane potential decreased linearly in both muscles during the ischemic period. The final lactate accumulation was higher and the decrease in membrane potential was less in the gastrocnemius than in the soleus. After release of the tourniquet both variables returned to normal or near normal values within 1.5 h in both muscles. A significant correlation was found between the lactate content and the membrane potential in both muscles during the entire experimental period. It is suggested that the depolarisation occurring in skeletal muscle during hypoxia is partly caused by changes in intracellular pH.

Hormonal and physiological responses to exercise in relation to the menstrual cycle.

The symposium presented in this issue is both an example and a result of the current interest and active research in the area of women and exercise. In the past few years, interest has focused on two overlapping areas: 1. the effect of exercise and training on menstrual function and exercise related amenorrhea 2. the effect of the menstrual cycle, with its varying basal concentrations of estradiol and progesterone, on the hormonal and physiological responses to exercise. This paper will address the latter area and will review our own work as well as that of others.

Physiology of diving in marine mammals.

Natural dives are usually short and aerobic, involving swimming effort that is not energetically costly. Present evidence indicates few, if any, major biochemical enzyme adaptations that would promote an extended anaerobic capacity in marine mammals. We have discussed the lack of evidence for unusual biochemical adaptations for anaerobiosis and the importance and characteristics of natural aerobic dives. What explanation can we offer for the known capability of some marine mammals, such as Weddell seals and sperm whales, to remain submerged for over an hour? Attributes that serve them well for shorter dives are important also for long dives, such as: (a) a large total oxygen store relative to body size, and (b) parsimonious use of blood oxygen due to the lowered energy requirements of various organs. During long dives, some organs deplete their oxygen stores, become predominately dependent on anaerobic metabolism, and build up large quantities of lactate. The result is a degree of fatigue that leaves the animal incapable of further intense diving activities for some time. Consequently, long anaerobic dives are quite rare in nature.

Studies on the formation of N6-hydroxylysine in cell-free extracts of Aerobacter aerogenes 62-1.

We have investigated conditions optimal for the conversion of L-lysine to its N6-hydroxy derivative by partially purified cell-free extracts of Aerobacter aerogenes 62-1. The enzyme system was highly specific to L-lysine: the D-isomer and, the N2- or N6-derivatives of lysine, and alpha-amino acids were not hydroxylated. Most of the latter compounds had little effect onthe hydroxylation of L-lysine. However, -l-glutamic acid and L-glutamine enhanced the hydroxylation, with half-maximal activation achieved at 100 micrometers concentration of the effector. The Km values for pyruvate and L-(+)-lactate (compounds known to stimulate N-hydroxylysine formation) were found to be approx. 100 micrometers. The data show that N-hydroxylation of the amino acid precedes acylation in the biosynthesis of hydroxamic acid in A. aerogenes 62-1.

[The effect of glucose substitutes on human uric acid metabolism (author's transl)]. / Wirkung von Glukoseaustauschstoffen auf den Harnsäurestoffwechsel des Menschen

Fructose, sorbitol and xylitol produce a rise in serum uric acid during intravenous injection. This is most pronounced with xylitol. The amount of increase in uric acid depends on the amount given. Irreversible decomposition of preformed adenine nucleotide in the liver is considered most important as a cause. Patients in whom there is already an impairment of the energy output on the part of the liver should not be given fructose or sorbitol and certainly not xylitol intravenously.