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Encephalopathy, oxygen consumption, visceral amino acid clearance, and mortality in cirrhotic surgical patients.

To assess the relationship of the high mortality of coma in cirrhotic surgical patients to defects in energy metabolism, reduced utilization of amino acids by the liver and other visceral tissues, oxygen consumption, central plasma clearance rate of amino acids (CPCR of amino acids), and the plasma concentrations of plasma inducing factors were measured in a series of 59 cirrhotic patients. They were classed as alert, encephalopathic, and comatose (Groups A, E, and C, respectively). The comatose group was set apart from the other two by a significantly higher mortality of 83 percent (p less than 0.005) combined with a lower whole body oxygen consumption of 103 +/- 6.8 ml/min per m2 compared with 135 +/- 10 ml/min per m2 in alert patients and 159 +/- 12 ml/min per m2 in the encephalopathic patients (p less than 0.01) and CPCR of amino acids of only 120 +/- 20 ml of plasma/min per m2 compared with 240 +/- 30 ml of plasma/min per m2 in the alert patients and 300 +/- 50 in the encephalopathic patients (p less than 0.01). An inverse correlation of tyrosine and phenylalanine concentrations existed with both whole body oxygen consumption (r = -0.56, p less than 0.01) and also with total amino acid clearance (r = -0.61, p less than 0.01). Tyrosine and phenylalanine concentrations also correlated directly with the octopamine concentration (r = 0.64, p less than 0.01). Thus, we conclude that coma is a symptom of hyperaminoacidemia, but that death is the result of impaired oxidative energy production and a deficiency of amino acid clearance for synthesis of proteins required for survival.

Adult↗

Determination of dopamine, norepinephrine, and related trace amines by prechromatographic derivatization with naphthalene-2,3-dicarboxaldehyde.

Dopamine, norepinephrine, octopamine, tyramine, and 3,4-dihydroxybenzylamine react readily with naphthalene-2,3-dicarboxaldehyde in the presence of cyanide ion under mild conditions to give highly fluorescent cyanobenz[f]isoindole (CBI) products. The CBI products exhibit good solution chemical stability. The high-fluorescence quantum efficiency of the CBI fluorophore and the ability to excite these adducts in the visible region (420-450 nm) enhance the sensitivity and selectivity of this derivatization detection technique. The CBI products of catecholamines and "trace" amines are readily separated by reverse-phase HPLC giving detection limits in the 20 to 60 fmol range (S/N = 3). A prechromatographic derivatization HPLC assay for the trace analysis of dopamine and norepinephrine in urine is described.

Biogenic Amines↗

The effect of dietary precursors on the excretion of amines and their metabolites in the rat.

The influence of diet on the excretion of catecholamines, some of their metabolites, and pHPG, an octopamine metabolite, was examined. Two groups of rats were fed either a cereal-containing standard laboratory Purina rat chow or a cereal-free casein diet. Use of the standard chow resulted in significant increases in the urinary values for total MHPG, pHPG, DHPE, MHPE, and free and total DOPAC by the seventh day in comparison to the casein diet. No changes were noted in the excretion of free and total NE, DA, and HVA. The data indicate that it is necessary to place the animals on the casein diet several days before determining the excretion of the aforementioned metabolites.

3,4-Dihydroxyphenylacetic Acid↗

The metabolism of L-m-tyrosine: the use of a putative precursor to investigate the increased production of m-hydroxymandelic acid in phenylketonuria.

Several urinary meta-substituted phenols appear to be of endogenous origin. However, the production of these compounds is reduced in phenylketonuria with the exception of m-hydroxymandelic acid whose excretion is approximately doubled. This phenomenon has been investigated in two patients with phenylketonuria using the putative precursor L-m-tyrosine labelled with deuterium. Metabolism of this compound in these patients was comparable to that in healthy adults although much less was converted to m-hydroxymandelic acid and the excretion pattern of this metabolite was different. This apparent anomaly is attributed to smaller metabolic compartments in phenylketonuria and a lower threshold for the metabolism of m-tyramine via beta-hydroxylation. Incorporation into the natural pathway was shown by depletion of endogenous m-hydroxymandelic acid. The results are further support for the ideas that the amine precursors of m-hydroxymandelic acid, m-tyramine and m-octopamine, have a functional role and may be important in the pathogenesis of phenylketonuria.

2-Hydroxyphenethylamine↗

Studies on the depletion of brain amines by m-tyrosine.

The biochemical basis of the well-known physiological and pharmacological actions of m-tyrosine was examined by a detailed study of its effect on the brain biogenic amines. m-Tyrosine was injected i.p. and rat brain monoamine levels were measured. Endogenous levels of dopamine, norepinephrine and serotonin all showed approximately 50% reductions 1 h after the administration of L-m-tyrosine at 150 mg/kg. These actions of L-m-tyrosine could be blocked by the inhibition of the central dopa decarboxylase. Depletion of brain monoamines was also observed with the D-isomer of m-tyrosine, although this effect was less pronounced than that of the L-isomer. In vitro experiments with rat brain homogenates showed that L-m-tyrosine, m-tyramine and m-octopamine enhanced in efflux of exogenous labeled monamines from brain particles, whereas D-m-tyrosine was completely ineffective. From these results it is concluded that the observed decreased in brain monamine levels by L-m-tyrosine may be due to a m-tyramine-enhanced release of the amines which are quickly metabolized in vivo.

Animals↗

Regulation of fat body mitochondrial respiration in Periplaneta americana by a novel factor from the corpus cardiacum.

Respiration of fat body (Periplaneta americana) mitochondria is increased by pretreatment of the tissue with corpus cardiacum (CC) extract. The magnitude of the increase depends on the type of substrate supplied for oxidation. With 5 mM pyruvate the respiration increased 22%, decreasing to 0 with 1 mM pyruvate. In contrast, 50 microM and 0.2 mM palmitic acid supported an increase in CC-stimulated respiration of 14 and 44%, respectively. Unlike crude CC extract, the synthetic hyperglycemic peptides CCI and CCII failed to alter the respiratory activity of fat body mitochondria. In common with the action of CC extract pretreatment of the fat body in vitro with 10(-5) M cyclic AMP, 10(-5) M 8-bromo-cyclic AMP, or 10(-5) M forskolin increased mitochondrial respiration approximately 30%. Octopamine (10(-4) M) elicited a response similar to that obtained with CC extract. Neither 10(-5) M cyclic AMP nor 10(-5) M 8-bromo-cyclic AMP stimulated respiration when applied directly to the mitochondria. These results suggest that the factor in CC extract manifests its effect intracellularly through the activation of a cyclic AMP-dependent protein kinase. This interpretation is also based on the finding that diamide, an inhibitor of protein kinase, inhibits CC-dependent and cyclic AMP-dependent mitochondrial respiration. The physiological role of the CC factor responsible is not known.

8-Bromo Cyclic Adenosine Monophosphate↗

Alterations of GABA-A and dopamine D-2 brain receptors in dogs with portal-systemic encephalopathy.

The binding characteristics of gamma-aminobutyric acid-A (GABA-A) receptors and the kinetic characteristics of the target enzyme of GABA synthesis in nerve terminals, glutamic acid decarboxylase (GAD), were studied in a dog model of portal-systemic encephalopathy obtained by porta-caval shunt performed in dimethylnitrosamine pretreated animals. Furthermore the properties of dopamine receptors and the levels of catecholamines of encephalopathic dogs were investigated. The mild stage of encephalopathy was characterized by an up-regulation of the inhibitory GABA-A receptors probably related to a decrese of GABA in nerve terminals since GAD was decreased and by a slight decrease of catecholamines and by an increased synthesis of octopamine associated with a decreased affinity of dopamine receptors. In the severe stage there was a selection of high affinity GABA-A receptors with an increased number of benzodiazepine recognition sites which were supersensitive to GABA stimulation, a decreased number of Dopamine D-2 receptors and a marked reduction of catecholamines. These data seem to suggest that the neurological disturbances of experimental portal-systemic encephalopathy might be the result of an imbalance between inhibitory and excitatory systems leading to a prevalence of the first one.

Animals↗

Two-way avoidance behavior of the roman rat strains following brain lesions.

Rats of the Roman strains, Roman High Avoidance (RHA), Roman Low Avoidance (RLA) and Roman Control Avoidance (RCA) were submitted to 5 conditioning sessions in a two-way shuttle box; the sessions were separated by a 24 hr-interval. Two indices were computed: a within-session progression index W and an inter-session progression index I. Index W was comparable in all three strains; index I was strongly negative in RLA and equal to zero or slightly positive in RHA rats. Lesion of the hippocampal commissure (HC) significantly facilitated CAR acquisition in RLA by making index I less negative: performances of RCA and RHA rats were unaffected. Lesions of the centro medianum parafascicularis nuclei of the thalamus (CM) significantly impaired acquisition of CAR in RCA and RHA by making index I more negative; performances of RLA rats were unaffected. Neither lesion altered index W. These results support the "freezing hypothesis" in the explanation of the Roman strain differences. They are in agreement with recent data on the emotional characteristics of the Roman strains and on the role of octopamine in avoidance behavior.

Animals↗

Amino acids and memory consolidation in the cricket. II: Effect of injected amino acids and opioids on memory.

The effect of injections of selected amino acids on memory, given before a maze-learning, was investigated. Thirsty crickets (Pteronemobius sp.) were trained to turn only to one side of a symmetrical Y-shaped maze using reinforcements with water. The insects retained the learned task 24 hr later. N2 anoxia applied immediately after training produced retrograde amnesia. Injections of Ala, Arg, Gln or morphine prior to training blocked the amnesic action of anoxia, whereas those of Cys, Met, Pro, Orn, octopamine or naloxone did not. Naloxone blocked long-term memory formation, but not learning, whereas Pro and Orn blocked both. The antiamnesic effect of morphine and Arg, but not that of Ala, was blocked by naloxone. A hypothesis assigning a neuromodulatory role to some amino acids is put forward.

Amino Acids↗

Antisera against catecholamines: specificity studies and physicochemical data for anti-dopamine and anti-p-tyramine antibodies.

Antibodies against dopamine and p-tyramine were raised in rabbits. The two catecholamines were conjugated to albumin by glutaraldehyde. The specificity of the antibodies was established by equilibrium dialysis competition experiments using an immunoreactive tritiated derivative synthesized by coupling dopamine or p-tyramine to N-alpha-acetyl-L-lysine N-methylamide with glutaraldehyde. Hence, these radiolabelled ligands mimicked the antigenic determinant of conjugated immunogens. A comparison of the data obtained showed the high specificity of each antiserum for its hapten coupled by glutaraldehyde. The anti-dopamine antibodies recognized dopamine-glutaraldehyde but not p-tyramine-glutaraldehyde. The opposite occurred for the anti-p-tyramine antibodies. A slight modification of the molecular structure provided the opportunity for a specific response against that molecule. But this difference was more important when related to the hapten region where the antibody affinity was maximal. The cross-reactivity was observed to be more important dopamine and p-tyramine than between dopamine and noradrenaline on the one hand and between p-tyramine and dopamine than p-tyramine and octopamine on the other hand.

Animals↗

Stimulatory effect of dopamine on acid secretion from the isolated rat stomach.

The effect of dopamine (DA) on acid secretion was studied using the everted preparation of isolated rat stomachs. DA concentrations, measured by HPLC-ECD in the rumen, corpus and antrum were 1.06 nmol/mg protein, 0.49 nmol/mg protein and 2.92 nmol/mg protein, respectively. DA stimulated acid secretion at a concentration of 10 nM and elicited the maximum response at 10 microM, which was at a level approximately 1.56-fold that of the spontaneous secretion but only about half that of secretion induced by histamine at a concentration of 0.3 mM. The concentration-dependent stimulation by DA was antagonized by octopamine and SCH 23390. Failure of proglumide and cimetidine to affect this stimulation ruled out the participation of histamine and/or gastrin. Scopolamine and tetrodotoxin completely inhibited the acid secretion induced by low concentrations of DA but inhibited only partially the response induced by high concentrations of DA. The results obtained indicate that DA induces acid secretion via activation of the dopamine D1 receptor, located on the cholinergic neurons and on some nonneuronal cells, in the rat stomach.

Animals↗

Neurotransmitter and neuropeptide modulation of high affinity choline uptake in Limulus brain.

The role of neurotransmitters in the modulation of the sodium-dependent high affinity choline uptake system (HAChUS) of the horseshoe crab, Limulus polyphemus has been investigated utilizing a tissue slice preparation. Choline uptake was significantly decreased by carbachol but unaffected by atropine and d-tubocurarine. The muscarinic agonist oxotremorine decreased choline uptake by 30.4% while the muscarinic antagonist, pirenzepine, increased uptake by 29.6%. Applied in combination, pirenzepine and oxotremorine abolished their individual effects resulting in control values for choline uptake. The non-cholinergic transmitters octopamine and serotonin significantly enhanced choline uptake. The neuropeptide proctolin elicited a 20% increase in choline transport whereas Phe-Met-Arg-Phe (FMRF) amide was without effect. This study demonstrates that neurotransmitters and neuropeptides modulate the HAChUS, possibly through specific receptor-mediated second messenger systems.

Animals↗

FMRF-NH2-like factor from neurohaemal organ modulates neuromuscular transmission in the locust.

YGGFMRFamide, FMRFamide and related peptides potentiate transmission at locust slow motor synapses. Since immunohistochemical evidence points to neurohaemal organs (NHO) as potential sources for endogenous RFamide-like peptides we have applied extracts from the metathoracic NHO, equivalent to 1/3 NHO, to the metathoracic extensor tibiae muscle. NHO-extract depolarizes the muscle fibre and increases its membrane resistance; enhances transmitter release; and increases the amplitude of contraction and the rate of relaxation. These effects are in good qualitative and quantitative agreement with those of YGGFMRFamide (5 X 10(-8) to 10(-7) M). Octopamine which partly acts like YGGFMRFamide cannot account for the NHO-effects.

Animals↗

Effects of octopaminergic substances on the labellar lobe spreading response in the blowfly Protophormia terraenovae.

The effectiveness of labellar taste input in evoking the feeding response of labellar lobe spreading in Protophormia was greatly increased by injecting flies with clonidine or DL-octopamine. This effect was suppressed when either drug was administered together with yohimbine; when injected alone, the latter determined a strong decrease in the feeding responses. Drug treatment did not determine variations in labellar taste input or evoke spontaneous activity in the denervated muscle. These results suggest that the injected substances lower the feeding-related reflex threshold at the CNS level.

Action Potentials↗

Effects of nialamide on responses of dog isolated arteries to tyramine and transmural electrical stimulation.

In dog mesenteric arteries, nialamide (10(-5) M), a monoamine oxidase inhibitor, potentiated contractile response to tyramine but not to noradrenaline or transmural electrical stimulation (TES). Bretylium, desipramine or prior reserpinization inhibited the potentiating action of nialamide on the tyramine-induced contraction. Contractions induced by octopamine were not reduced by prior reserpinization. In the coronary artery, relaxing responses to tyramine were potentiated but those to noradrenaline and TES were not potentiated by nialamide. In the cerebral artery, nialamide failed to potentiate tyramine-induced contraction. The functional role of intraneuronal monoamine oxidase in sympathomimetic effects is discussed.

Animals↗

Formamidine-mediated inhibition of rat platelet aggregation.

The formamidine pesticide chlordimeform (CDF) was a strong inhibitor of aggregation of rat platelets induced by collagen and arachidonic acid and was a weak inhibitor of that induced by ADP. With the exception of 4-chloro-o-formotoluidide which was an inhibitor of arachidonic acid-induced aggregation only, the CDF metabolites were without discernible effect on aggregation induced by these agents. Amitraz, another formamidine pesticide, inhibited arachidonic acid-induced aggregation but was without effect on that induced by collagen or ADP. Inhibition of collagen- and/or arachidonic acid-induced aggregation by formamidines was concentration-dependent. Although platelets underwent shape change, primary aggregation was markedly inhibited, and secondary aggregation was abolished in some cases. CDF, its two N-desmethyl metabolites, and octopamine, but not amitraz, caused significantly elevated levels of cyclic AMP in platelet rich plasma as compared to controls; however, this effect did not fully account for the action of formamidines on aggregation.

Adenosine Diphosphate↗

Actions of noradrenaline and some other biogenic amines on the body-wall muscles of an echiuroid, Urechis unicinctus.

1. Noradrenaline inhibited contractions of the body-wall strips of U. unicinctus in response to electrical pulse of stimulation and L-proline, while serotonin enhanced them. In addition, serotonin increased the rate of relaxation of twitch contraction. Octopamine enhanced the contractions but was less potent than serotonin. 2. Acetylcholine-contraction of the isolated inner circular body-wall muscle was also inhibited by noradrenaline and enhanced by serotonin. 3. Phentolamine, an alpha-adrenoreceptor blocker, potentiated tetanic contraction of the strip, though it did not alter twitch contraction. The inhibitory action of noradrenaline was blocked by phentolamine. 4. Noradrenaline hyperpolarized the fibre membrane of the inner circular muscle, while serotonin did not alter the membrane potential. The action of noradrenaline was blocked by phentolamine. 5. Bioassay of the body-wall extract and the ventral nerve-cord extract suggests that these extracts contain serotonin-like and noradrenaline-like substances, respectively. 6. These results suggest that noradrenaline and serotonin may be neurotransmitters or neurohormones modulating muscle contraction in the body wall of U. unicinctus.

Adrenergic alpha-Antagonists↗

Biogenic amines: their occurrence, biosynthesis and metabolism in the locust, Schistocerca gregaria, by gas chromatography-negative-ion chemical ionisation mass spectrometry.

Extraction-derivatisation techniques have been developed for the unambiguous identification of biogenic amines, and their putative amino acid precursors and metabolites (both major and minor), in single ventral thoracic nerve cords of the locust. Schistocerca gregaria, by the use of gas chromatography-negative-ion chemical ionisation mass spectrometry with selected ion monitoring. In addition the configuration of that enantiomer of p-octopamine present in the thoracid nervous system of the locust was established as R using the chiral derivatisation reagent, (-)-heptafluorobutyrylphenylalanyl chloride.

Acetylation↗