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The influence of ammonia, biogenic amines and gamma-aminobutyric acid on grass silage intake in sheep.

We investigated whether biogenic amines alone, or a combination of NH3, amines and gamma-aminobutyric acid (GABA) influenced grass-silage intake, intake behaviour and rumen liquid content in sheep. Three diets were studied: a grass silage preserved with formic acid (4 litres/tonne) (FAS), FAS with 4.9 g amines/kg DM added (FAS+A), and FAS supplemented with a combination of N-components at the following concentrations: 2.7 g amines, 3.0 g NH3 and 5.0 g GABA/kg DM (FAS+C). The diets were offered ad libitum, once daily to six rumen-cannulated Texel wethers in a crossover design. Daily DM intake (DMI; g DM/d) tended to be influenced by diet (P = 0.08). The DMI of FAS+A was similar to that of FAS alone, whereas that of FAS+C tended to be higher. The mean rate of ingestion (g DM/min) over all feeding bouts tended to be the lowest for FAS+A (P = 0.06). No differences were found among the diets concerning intake behaviour during the principal meal. Average intake rate of the small meals tended to be the lowest for FAS+A (P = 0.06). Although rumen NH3 concentration was higher (P < 0.05) after the principal meal, rumen pH, osmolality, rumen pool size and liquid content were not significantly altered by adding amines or the mixture of N-components to FAS. We conclude that biogenic amines or N-containing products of protein fermentation in concentrations normally found in poor-quality silages do not reduce the intake of well-preserved formic acid-treated silage. A direct effect on chemostatic regulation of intake was not observed, but a slight negative effect on silage palatability cannot be excluded.

Ammonia↗

Investigation of the presence of biogenic amines and ethyl carbamate in kenkey made with maize and maize-cowpea mixtures as influenced by process conditions.

Kenkey is a fermented and cooked maize dough from Ghana. The effect of manufacturing conditions, i.e. fermentation and cooking, and of protein-enrichment by cowpea addition (20% of total weight) on the occurrence of toxic microbial products, namely biogenic amines and ethyl carbamate, were investigated. The levels of biogenic amines in all-maize kenkey were very low (total amines < 60 ppm), but were significantly increased by addition of red cowpea (total amines < 200 ppm, mainly cadaverine and tyramine), and even more by white cowpea (total amines < 500 ppm, mainly putrescine and tyramine). Histamine was absent (< 5 ppm) in all samples. The effects of fermentation and cooking were less pronounced than the influence of cowpea addition. Prolonged cooking of kenkey resulted in lower levels of putrescine, but did not significantly reduce tyramine levels. Ethyl carbamate levels were negligible (< 11 ppb) in all treatments.

Biogenic Amines↗

Effect of the trichothecene deoxynivalenol on brain biogenic monoamines concentrations in rats and chickens.

Male Sprague-Dawley rats (180 g) and 28-day-old Single Comb White Leghorn Cockerels (300 g) were orally dosed with deoxynivalenol (DON) at 2.5 mg kg-1 body weight. In the first experiment, whole brains were collected at 2, 6, 12, 24 and 48 hours after the toxin treatment and analyzed for brain biogenic monoamines by high-performance liquid chromatography with electrochemical detection. Although several interesting trends were observed, DON did not influence whole brain concentrations of monoamine neurotransmitters or their metabolites in either species, at any time. In a second experiment, brains were collected 24 hours postdosing, dissected into 5 brain regions (pons and medulla oblongata, cerebellum, hypothalamus, hippocampus and cerebral cortex), and analyzed. DON treatment resulted in significantly elevated concentrations of serotonin (HT) and 5-hydroxyindole-3-acetic acid (HIAA) in all brain regions of the rat. However, this was not seen in poultry, where DON treatment resulted in a decrease in norepinephrine (NE) in the hypothalamus and hippocampus, and a decrease in dopamine (DA) in the pons and medulla oblongata region. These results suggest that DON influences brain biogenic amine metabolism, and that there may be intraspecies differences in the central effects of this mycotoxin.

Animals↗

Biogenic amines in cultured neuroblastoma and astrocytoma cells.

The presence of biogenic amines in cultured cells of mouse neuroblastoma C-1300 (clone NB-2a) was suggested by fluorescence-microscope histochemistry. Incubation in media containing L-[(14)C]tyrosine and L-[(14)C]tryptophan for 24 h, followed by high-voltage electrophoresis, radiochromatogram scanning, and scintillation counting, confirmed the presence of [(14)C]dopamine, [(14)C]norepinephrine, [(14)C]epinephrine, [(14)C]serotonin, [(14)C]tyramine, and [(14)C]octopamine. Dopamine, norepinephrine, epinephrine, and serotonin were demonstrated spectrophotofluorometrically in concentrations, expressed as micrograms amine per milligram protein, of 1.19, 0.027, 0.038, and 0.148, respectively, for cells in a stationary growth phase. Fluorescence-microscope histochemistry also suggested the presence of biogenic amines in cultured astrocytoma cells (cell line C6). Spectrophotofluorometric assay of cells in a stationary growth phase demonstrated intracellular dopamine, norepinephrine, epinephrine, and serotonin in concentrations significantly lower than those of neuroblastoma cells.

Animals↗

The effects of acetylcholine and dopamine on the caudate nucleus depleted of biogenic amines.

Because it has been proposed that the reduction of the striatal biogenic amines in Parkinson's disease leads to an imbalance between the actions of acetylcholine and dopamine, we have studied the effects of these substances, liberated from multibarrelled micropipettes, on the firing of single neurons in the feline caudate nucleus depleted of biogenic amines by long-standing nigrostriatal lesions. Compared with neurons in intact cats, those in cats with lesions were more easily excited by acetylcholine and less easily supressed by dopamine. These results suggest that the depletion of the striatal amines decreases the neuronal susceptibility to dopamine and increases that to acetylocholine, possibly by changing the sensitivity or the number of the neuronal receptors of these agents.

Acetylcholine↗

Biogenic amines in the taste organ.

The presence and content of biogenic amines in taste disk-bearing fungiform papillae of the frog, Rana esculenta, the only available model of an isolated taste organ, were verified by means of HPLC. Fungiform papillae were found to contain measurable amounts of serotonin, epinephrine and norepinephrine. The amounts of serotonin and epinephrine were significantly higher in fungiform papillae than in the general mucosa of the tongue. Moreover, the epinephrine content of fungiform papillae was found to differ across the tongue, in accordance with previous physiological studies showing an inhomogeneous response of different tongue regions to taste stimuli. Ultrastructural and histochemical investigations confirmed the presence of catecholamine and serotonin. The latter was found to be contained mainly in the basal cells of the frog taste disk. These results extend previous qualitative data on the presence of biogenic amines in taste chemoreceptors.

Animals↗

The non-enzymatic hydrolysis of oligoribonucleotides VI. The role of biogenic polyamines.

Single-stranded oligoribonucleotides containing UA and CA phosphodiester bonds can be hydrolyzed specifically under non-enzymatic conditions in the presence of spermidine, a biogenic amine found in a wide variety of organisms. In the present study, the rate of oligonucleotide and tRNA(i)(Met)hydrolysis was measured in the presence of spermidine and other biogenic amines. It was found that spermine [H(3)N(+)(CH(2))(3)(+)NH(2)(CH(2))(4)(+)NH(2)(CH(2))(3)(+)NH(3)] and putrescine [H(3)N(+)(CH(2))(4)(+)NH(3)] can replace spermidine [H(3)N(+)-(CH(2))(4)(+)NH(2)(CH(2))(3)(+)NH(3)] to induce the hydrolysis. For all three polyamines, a bell-shaped cleavage rate versus concentration relationship was observed. The maximum rate of hydrolysis was achieved at 0.1, 1.0 and 10 mM spermine, spermidine and putrescine, respectively. Moreover, we found that the hydrolysis requires at least two linked amino groups since two aminoalcohols, 2-aminoethanol and 3-aminopropanol, were not able to induce the cleavage of the phospho-diester bond. The optimal cleavage rate of the oligo-ribonucleotides was observed when amino groups were separated by tri- or tetramethylene linkers. The methylation of the amino groups reduced the ability of diamines to induce oligoribonucleotide hydrolysis. Non-enzymatic cleavage of tRNA(i)(Met)from Lupinus luteus and tRNA(i)(Met)from Escherichia coli demonstrate that both RNAs hydrolyze as expected from principles derived from oligoribonucleotide models.

Biogenic Polyamines↗

Cerebrospinal fluid biogenic amine metabolites, plasma-rich platelet serotonin and [3H]imipramine reuptake in the primary fibromyalgia syndrome.

BACKGROUND: Primary fibromyalgia syndrome (PFS) is a chronic disorder commonly seen in rheumatological practice. The pathophysiological disturbances of this syndrome, which was defined by the American College of Rheumatology in 1990, are poorly understood. This study evaluated, in 30 patients, the hypothesis that PFS is a pain modulation disorder induced by deregulation of serotonin metabolism. OBJECTIVES: To compare platelet [(3)H]imipramine binding sites and serotonin (5-HT) levels in plasma-rich platelets (PRP) of PFS patients with those of matched healthy controls and to compare the levels of biogenic amine metabolites in the cerebrospinal fluid (CSF) of PFS patients with those of matched controls. METHODS: Platelet [(3)H]imipramine binding sites were defined by two criteria, B(max) for their density and K(d) for their affinity. PRP 5-HT and CSF metabolites of 5-HT (5-hydroxyindoleacetic acid, 5-HIAA), norepinephrine (3-methoxy, 4-hydroxy phenylglycol, MHPG) and dopamine (homovanillic acid, HVA) were assayed by reversed-phase high-performance liquid chromatography with coulometric detection. RESULTS: [(3)H]Imipramine platelet binding was similar (P=0.43 for B(max) and P=0.30 for K(d)) in PFS patients (B(max)=901+/-83 fmol/mg protein, K(d)=0.682+/-0.046) and in matched controls (B(max)=1017+/-119 fmol/mg protein, K(d)=0.606+/-0.056). PRP 5-HT was significantly higher (P=0.0009) in PFS patients (955+/-101 ng/10(9) platelets) than in controls (633+/-50 ng/10(9) platelets). When adjusted for age, the levels of all CSF metabolites were lower in PFS patients. The CSF metabolite of norepinephrine (MHPG) was lower (P:=0.003) in PFS patients (8.33+/-0.33 ng/ml) than in matched controls (9.89+/-0.31 ng/ml) and 5-HIAA was lower (P=0.042) in PFS female patients (22.34+/-1.78 ng/ml) than in matched controls (25.75+/-1.75 ng/ml). For HVA in females, the difference between PFS patients (36.32+/-3.20 ng/ml) and matched controls (38.32+/-2.90 ng/ml) approached statistical significance (P=0.054). CONCLUSION: Changes in metabolites of CSF biogenic amines appear to be partially correlated to age but remained diagnosis-dependent. High levels of PRP 5-HT in PFS patients were associated with low CSF 5-HIAA levels in female patients but were not accompanied by any change in serotonergic uptake as assessed by platelet [(3)H]imipramine binding sites. These findings do not allow us to confirm that serotonin metabolism is deregulated in PFS patients.

Biogenic Amines↗

Biogenic amines and depression (introduction to symposium).

The involvement of biogenic amines--primarily noradrenaline (NA) and serotonin (5-HT)--in depressive disorders was suggested by the finding more than twenty years ago that imipramine possessed antidepressant properties and concomitantly altered the availability of NA in the brain. Since then, numerous antidepressants closely related chemically and pharmacologically to imipramine have been introduced. It has generally been assumed that these "tricyclics" alleviate depression by influencing the neurotransmitters (NA, 5-HT) at crucial receptor sites in the brain. Thus, the "biogenic amine hypothesis of depression" postulates that depression is due to a reduced functional activity of one or more brain amines. The tricyclics appear to be antidepressants due to their inhibition of the neuronal reuptake of NA in the brain. A decade ago, Carlsson, Coppen, Lapin, van Praag and other suggested that compounds selectively affecting the 5-HT uptake mechanism might be more effective antidepressants. This symposium will describe the clinical and preclinical results with zimelidine--the first 5-HT-selective uptake inhibitor.

Antidepressive Agents↗

Predominant cytosolic distribution of serotonin in rat pineal gland in contrast to biogenic monoamine localization in midbrain and adrenal gland.

The subcellular distribution of serotonin and norepinephrine in the rat pineal gland was studied by tissue fractionation and compared with that of biogenic monoamines in the adrenal gland and midbrain. Homogenized tissues were fractionated by ultracentrifugation or by filtration through cellulose ester membranes. Most of the epinephrine (70-80%) and norepinephrine (62-82%) present in the adrenal glands was detected in the particulate fraction. The same distribution was found for serotonin (68.5%) and norepinephrine (59%) in the midbrain and for norepinephrine (62.5%) in the pineal gland. However, most of the serotonin in the pineal was found in the soluble fraction (89.5-98%). This suggests that the great majority of serotonin in the rat pinealocytes is cytosolic and thus is not stored in subcellular vesicles, in contrast to the biogenic monoamines in the midbrain or adrenal gland.

Adrenal Glands↗

Tetrahydrobiopterin and biogenic amine metabolism in neuropsychiatry, immunology, and aging.

Tetrahydrobiopterin (BH4) is essential for biogenic amine synthesis, and alterations in its metabolism occur at birth (atypical PKU), in neuropsychiatric illnesses, and in aging. BH4 therapy has been attempted in atypical PKU and in neuropsychiatric illness with some success and may become more viable as more is learned about BH4 metabolism and ways are discovered to elevate brain BH4 levels. It is intriguing to consider that a genetic defect in BH4 biosynthesis occurring at birth might go unrecognized and contribute to altered biogenic amine metabolism that occurs in neuropsychiatric illness. Since there seems to be a sensitivity of BH4 metabolism to genetic alterations, it is possible that altered BH4 metabolism is involved in some of deleterious effects associated with the aging process. A link between genetic alterations in BH4 metabolism at birth and adult neuropsychiatric illness and aging remains to be established, although this seems plausible. The presence of BH4 and other pterins in cells of the immune system as well as the pineal gland and other neuroendocrine tissues suggests the potential for other functions of pterins. Hopefully, future research will uncover the full potential for the therapeutic use of BH4 in a variety of diseases as well as elucidating other potential roles for pterin molecules which are present in many different systems.

Aging↗

Biogenic amine synthesis defect in dihydropteridine reductase deficiency.

In the enzymatic hydroxylation of aromatic amino acids, tetrahydrobiopterin is the essential cofactor. Regeneration of tetrahydrobiopterin requires dihydropteridine reductase, without which there should be a deficiency of hydroxylated amino acids and their products, biogenic amines. Assay of biopsied brain cortex of a patient with a deficiency of dihydropteridine reductases showed low concentrations of serotonin and dopamine, and this was reflected in the concentrations of their major metabolites measured in cerebrospinal fluid from lumbar, ventricular, and subarachnoid spaces. The metabolite concentrations were restored to normal, or above normal, by treatment with specific amino acids which bypass the metabolic block at the hydroxylation step. It is postulated that the seizures and neurological deterioration of the patient were related to a deficiency in the synthesis of biogenic amine neurotransmitters.

5-Hydroxytryptophan↗

Effects of chlordiazepoxide administration on biogenic amines in cat brain.

Cats underwent treatment with chlordiazepoxide hydrochloride (0.4, 10.0, and 20.0 mg/kg per os), for 7 consecutive days, and were killed 18 h after the last administration. The endogenous levels of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), noradrenaline (NA), and dopamine (DA) were assayed in 12 brain areas. Few effects on 5-HT, 5-HIAA, and NA content and on the 5-HT:5-HIAA ratio were observed with a 0.4 mg/kg treatment. These changes were localized in the piriform lobe (amygdala), hippocampus, mesencephalon, and mesencephalon raphe nuclei. Moreover, the DA concentration was not affected. The changes produced by 10.0 and 20.0 mg/kg chlordiazepoxide treatments were extended to many more structures, including the limbic system, brainstem, diencephalon, and neostriatum with respect to 5-HT, 5-HIAA, and NA content and also to DA levels. The changes observed after the three doses generally included an increased 5-HT content, a decreased 5-HIAA level, a high 5-HT:5-HIAA ratio, and increased NA and DA concentrations. However, in some structures, a decreased NA content and an increased 5-HIAA level were found. The present results suggest that administration of chlordiazepoxide for 7 consecutive days in cats produces regional changes in the content of endogenous biogenic amines in the central nervous system (CNS) at low doses; much more extended effects are produced at high doses. These findings are in agreement with a reducing effect of benzodiazepines on the turnover and release of biogenic amines in the CNS, but also suggest that certain discrete areas are more involved in these changes, thus dissociating them from the rest of the brain.

Animals↗

Pulmonary vascular reactivity after repetitive exposure to selected biogenic amines.

The present study investigated the effects of repetitive exposure to a select group of biogenic amines (epinephrine, norepinephrine, histamine, and serotonin) on pulmonary vascular reactivity by constructing and analyzing a set of four sequential cumulative dose-response curves to one biogenic amine in the isolated blood-perfused left lower lobe of the cat lung in vivo. The dose-response curves were obtained under conditions of constant flow, insuring that the observed pressure changes in the lobe were pressor responses resulting from vasoconstriction rather than flow-related changes. Histamine and epinephrine demonstrated a progressive loss of initial vasoconstrictor activity, whereas the responses to serotonin remained unchanged after repetitive exposure. Norepinephrine demonstrated two different patterns of response, depending on the dose range employed; norepinephrine (0.068-2.27 nmol/ml) demonstrated a loss of the original vasoconstrictor activity, in a pattern similar to histamine and epinephrine, while higher doses of norepinephrine (0.34-9.1 nmol/ml) demonstrated no change in activity with a left shift in the concentration at which the maximal responses occurred, suggesting an increase in sensitivity as a result of repeated exposure. These results were obtained in the absence of significant alterations of arterial blood gases, changes in base-line tone in the experimental left lower lobe, or the development of severe pulmonary edema. These data suggest that only the agents that are capable of stimulating antagonistic vasoconstrictor and vasodilator receptors demonstrated a loss of pulmonary vasoconstrictor activity, which may result from a functional shift in the balance of antagonistic receptor activity with continued exposure.

Animals↗

Metamodulation of the biogenic amines: second-order modulation by steroid hormones and amine cocktails.

An evolutionarily conserved feature of neural systems is that they can be modified by neuromodulators. These modulatory chemical signals include the biogenic amines, octopamine (OA), serotonin (5-HT) and dopamine (DA). Such modulation effectively broadens the operational range in which specific neural circuits can function adaptively. This report discusses how these amines are themselves modulated; for example, by the steroid hormone 20-hydroxyecdysone (20-E) or by the addition of a second biogenic amine. Such second-order neuromodulation, termed metamodulation, is discussed in the context of two well-studied invertebrate systems: the tobacco hornworm moth Manduca sexta, a model of neurodevelopment and plasticity, and the medicinal leech Hirudo medicinalis, a long-favored preparation used to study neural circuits at the level of identified neurons. A portion of this article reviews our previous research of M. sexta that shows that the 'preadult' rise in 20-E is both necessary and sufficient for the increased levels of octopamine observed in the adult. Such elevated levels likely play an important role in the production and modulation of adult behaviors. The somatic growth of median octopaminergic neurons and the late expression of OA-immunoreactivity by novel lateral neurons are also demonstrated to be dependent on 20-E. New immunocytochemical results of stained dopaminergic neurons in the larval and adult moth brain are provided as well, and the potential influence of 20-E on the developmental expression of this neuromodulator is presented. Turning attention to the leech, data indicate that the actions of OA are dramatically altered when 5-HT is combined with OA in the bath surrounding the isolated nervous system. Although either OA or 5-HT alone induces fictive swimming behavior, a cocktail of these two amines strongly inhibits the generation of swimming. Subsequent removal of such a mixture induces nearly continuous swimming and constitutes the best swim-inducing stimulus encountered to date. To understand better how these nonadditive effects are achieved, new results are discussed that indicate that the leech brain is the target of metamodulation by the two amines. Both the arthropod and annelid systems presented here highlight the multiple levels of metamodulation that can exist in nervous systems, and the diverse ways that a modulator's actions can become altered over short or long time periods.

Animals↗

Studies of the influence of biogenic amines on central nervous system ischemia.

Several serotonin inhibitors have been shown to reduce neurologic deficits in experimental CNS ischemia. Using biochemical and histological methods we tested the effects of the serotonin inhibitors cyproheptadine and brom-LSD in a highly reproducible rabbit spinal cord ischemia model. Detailed mapping of regional spinal cord blood flow was used to guide sampling for the biochemical studies. We found that it is possible to study biochemical and morphological aspects of spinal cord ischemia in great detail using a combination of quite precise techniques. However, at this level of resolution there were no substantial changes in biogenic amine concentrations in severely ischemic or marginally perfused tissue after the durations of ischemia that cause the onset of irreversible tissue damage. Treatment with doses of serotonin inhibitors that produce preservation of neurological function did not cause significant alterations of tissue concentrations of biogenic amines or tissue morphology in treated versus untreated animals.

Animals↗

A method for counterstaining tissues in conjunction with the glyoxylic acid condensation reaction for detection of biogenic amines.

A method is described for counterstaining tissue for use with the glyoxylic acid reaction for visual detection of biogenic amines. Counterstaining is achieved by addition of the fluorescent dye malachite green to the sucrose-phosphate-glyoxylic acid (SPG) solution used for processing of cryostat sections of unfixed tissues. When bound to tissues, the dye provides red-orange fluorescence of background tissue, which contrasts well with the green to yellow fluorescence induced by the glyoxylic acid reaction product formed with biogenic amines. The counterstaining technique is demonstrated in a number of catecholamine-containing peripheral tissues and is compared to sections that were processed without counterstaining.

Adrenal Glands↗