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Cerebrospinal fluid monoamine metabolites in 114 healthy individuals 18-88 years of age.

Concentrations of the monoamine metabolites homovanillic acid (HVA), 5-hydroxy-indoleacetic acid (5-HIAA) and 4-hydroxy-3-methoxyphenylglycol (HMPG) were determined in lumbar cerebrospinal fluid (CSF) of 114 healthy individuals, 18-88 years of age, without histories, symptoms or signs of central nervous system dysfunction. The mean values (+/- SD) were 253 +/- 109 nmol/l for HVA, 125 +/- 54 nmol/l for 5-HIAA, 47 +/- 10 nmol/l for HMPG, and 2.10 +/- 0.52 for the HVA/5-HIAA ratio. Analyses of confounding factors revealed that all metabolites correlated negatively with body height, the values being lower in taller than in shorter individuals. This is probably attributable to a larger surface area for monoamine metabolite transport from the subarachnoid space in taller than in shorter individuals. These correlations make statistical adjustment for body height important in analyses of monoamine metabolite levels. Without considering body height, all monoamine metabolites showed a positive correlation with age, and higher levels of HVA and 5-HIAA were found in women than men. After statistical adjustment for the influence of body height, no differences in CSF monoamine metabolites levels were found between the sexes, and only 5-HIAA showed a positive correlation with age. There were no significant seasonal variations for any of the monoamine metabolites.

Adolescent↗

Cerebrospinal fluid kynurenic acid in male and female controls - correlation with monoamine metabolites and influences of confounding factors.

The concentrations of the tryptophan metabolite kynurenic acid (KYNA) and the monoamine metabolites homovanillic acid (HVA), 5-hydroxy-indoleacetic acid (5-HIAA) and 4-hydroxy-3-methoxyphenylglycol (HMPG) were determined in the cerebrospinal fluid (CSF) from 43 healthy volunteers (30 males and 13 females). Healthy female controls displayed higher CSF concentration of KYNA (1.91nM+/-0.20) compared to healthy male controls (1.06nM+/-0.07) and lower CSF levels of HMPG (39.2nM+/-2.0 and 43.4+/-1.2, respectively). CSF levels of HVA and 5-HIAA did not differ between females (181.3nM+/-21.9 and 93.7nM+/-11.4, respectively) and males (138.9nM+/-12.6 and 74.8nM+/-5.9, respectively). Positive intercorrelations were found between CSF KYNA, HVA and 5-HIAA while CSF content of HMPG did not correlate with KYNA or the other monoamine metabolites in CSF. A negative correlation was found between back length and CSF concentrations of KYNA, HVA and 5-HIAA and also between CSF KYNA levels and body height. The results of the present study suggest that concentrations of KYNA and the monoamine metabolites in CSF from healthy controls are dependent on gender and back length, which must be taken in consideration when analysing mixed groups of men and women. The higher KYNA concentration found in female controls compared to male might be attributed to a shorter back in women compared to men. Furthermore, these findings suggest that increased KYNA formation is associated with an increased dopamine and serotonin turnover.

Adult↗

Malignant gastric neuroendogrinomas. Ultrastructural and biochemical characterization of their secretory activity.

In two cases of malignant gastric tumors originally diagnosed as undifferentiated carcinomas electron microscopy revealed a neurosecretory type of granule. Subsequently tumor extracts were tested by biochemical methods and shown to have vanillylmandelic acid and 5-hydroxy-3-indoleacetic acid activity. Neither patient had signs or symptoms referable to the presence of these or related substances. These observations parallel those made in a variety of neuroendocrine tumors in which demonstration of neurosecretory granules or isolation of amine or peptide materials or their metabolites has not necessarily been reflected in clinical hormonal syndromes. Our findings indicate that regardless of clinically apparent hormonal activity or lack thereof, some undifferentiated gastric carcinomas may in fact derive from neuroendocrine APUD elements.

Aged↗

Effect of chronic protein restriction on motor co-ordination and brain neurotransmitters in albino rats.

In this study we evaluated the motor co-ordination in Wistar strain albino rats that were maintained on a protein-restricted diet for a period of 1 year immediately after the weaning period, by substituting 75% of the normal diet with a carbohydrate-rich diet deficient in protein, for a period of 1 year immediately after the weaning period. This type of chronic protein restriction caused disturbances in motor co-ordination. It also caused a significant reduction in the basal levels of dopamine, norepinephrine, epinephrine and serotonin along with their metabolites homovanillic acid (HVA), vanillyl mandelic acid (VMA) and 5-hydroxy indoleacetic acid (5HIAA) and precursor L-dopa in the corpus striatum and cerebellum. Changes in these neurotransmitters could have caused altered co-ordination in the protein-restricted animals.

Animals↗

Mathematical analysis of the chemosmotic polar diffusion of auxin through plant tissues.

Equations have been developed to describe the diffusional movement of a weak acid such as the auxin indoleacetic acid through a long file of vacuolated cells, where cellular accumulation is driven by the pH gradients across the cell membranes. If the permeability to the auxin anion is greater at one end of the cell than at the other, diffusional movement takes the form of polar transport, which exhibits: a nearly constant velocity either for the front or for a pulse of radioactive auxin, the capacity to move auxin against an external gradient of concentration, and a polar ratio that increases exponentially with the length of the section. The determinants of velocity include both diffusion through the vacuole and permeation steps at the cell membranes. Except for the permeabilities of the membranes to the anion, values are now available for all of the physical parameters in the equations. With reasonable estimates of permeability coefficients for the anion, the equations predict a velocity of transport of about 1 cm hr(-1), which agrees well with measured values. The analysis indicates, however, that the underlying cellular polarity may be greater than has been heretofore assumed. We thus demonstrate that the hypothesis of chemosmotic polar diffusion is capable of accounting quantitatively for the major features of auxin transport and provides a theoretical framework whose elements can be tested in future experiments.

Journal Article↗

Effect of age on cerebrospinal fluid levels of metabolites of biopterin and biogenic amines.

We identified an age effect on biopterin and biogenic amine metabolites in the cerebrospinal fluid from 56 neurologically normal patients, aged 1 mo to 80 y. The levels of total and reduced forms of biopterin, homovanillic acid, and 5-hydroxyindoleacetic acid were found to peak during the first year of life. The levels of these metabolites then gradually decreased, plateauing at the age of 20 y. We found significant correlations between biopterin, homovanillic acid, and 5-hydroxy-indoleacetic acid.

Adolescent↗

The Role of auxin, pH, and stress in the activation of embryogenic cell division in leaf protoplast-derived cells of alfalfa.

Culturing leaf protoplast-derived cells of the embryogenic alfalfa (Medicago sativa subsp. varia A2) genotype in the presence of low (1 microM) or high (10 microM) 2, 4-dichlorophenoxyacetic acid (2,4-D) concentrations results in different cell types. Cells exposed to high 2,4-D concentration remain small with dense cytoplasm and can develop into proembryogenic cell clusters, whereas protoplasts cultured at low auxin concentration elongate and subsequently die or form undifferentiated cell colonies. Fe stress applied at nonlethal concentrations (1 mM) in the presence of 1 microM 2,4-D also resulted in the development of the embryogenic cell type. Although cytoplasmic alkalinization was detected during cell activation of both types, embryogenic cells could be characterized by earlier cell division, a more alkalic vacuolar pH, and nonfunctional chloroplasts as compared with the elongated, nonembryogenic cells. Buffering of the 10 microM 2,4-D-containing culture medium by 10 mM 2-(N-morpholino)ethanesulfonic acid delayed cell division and resulted in nonembryogenic cell-type formation. The level of endogenous indoleacetic acid (IAA) increased transiently in all protoplast cultures during the first 4 to 5 d, but an earlier peak of IAA accumulation correlated with the earlier activation of the division cycle in embryogenic-type cells. However, this IAA peak could also be delayed by buffering of the medium pH by 2-(N-morpholino)ethanesulfonic acid. Based on the above data, we propose the involvement of stress responses, endogenous auxin synthesis, and the establishment of cellular pH gradients in the formation of the embryogenic cell type.

2,4-Dichlorophenoxyacetic Acid↗

Influence of Auxin and Gibberellin on in Vivo Protein Synthesis during Early Pea Fruit Growth.

Developing pea fruits (Pisum sativum L.) offer a unique opportunity to study growth and development in a tissue that is responsive to both gibberellins (GAs) and auxin (4-chloroindole-3-acetic acid[4-CI-IAA]). To begin a molecular analysis of the interaction of GAs and auxins in pea fruit development, in vivo labeling with [35S]methionine coupled with two-dimensional gel electrophoresis were used to characterize de novo synthesis of proteins during gibberellic acid (GA3)-, 4-CI-indoleacetic acid-, and seed-induced pea pericarp growth. The most significant and reproducible polypeptide changes were observed between molecular weights of 20 and 60. Comparing about 250 de novo synthesized proteins revealed that seed removal changed the pattern substantially. We identified one class of polypeptides that was uniquely seed induced and five classes that were affected by hormone treatment. The latter included 4-CI-IAA-induced, GA3-induced, GA3- and 4-CI-IAA-induced, 4-CI-IAA-repressed, and GA3- and 4-CI-IAA-repressed polypeptides. Similar patterns of protein expression were associated with both hormone treatments; however, changes unique to GA3 or 4-CI-IAA treatment also indicate that the effects of GA3 and 4-CI-IAA on this process are not equivalent. In general, application of 4-CI-IAA plus GA3 replaced the seed effects on pericarp protein synthesis, supporting our hypothesis that both hormones are involved in pea pericarp development.

Journal Article↗

Gas-liquid chromatography of trimethylsilyl derivatives of abscisic Acid and other plant hormones.

This paper describes a new method for qualitative and quantitative assay of abscisic acid and other acidic plant hormones, such as indoleacetic acid and the gibberellins, by the gas-liquid chromatography of their trimethylsilyl derivatives. Interfering substances in plant extracts were largely removed by preliminary column chromatography with carbon-celite and elution of the abscisic acid with 60% acetone, permitting direct determination of abscisic acid by gas-liquid chromatography using a flame ionization detector. (A level of 0.65 mg/kg fr wt was found.) This method enables measurement of amounts of abscisic acid as low as 0.025 mug. In impure samples collected by gas-liquid chromatography the abscisic acid recovered could be measured quantitatively by use of its ultraviolet absorption maximum at 260 mmu.

Journal Article↗

CSF sulfatide distinguishes between normal pressure hydrocephalus and subcortical arteriosclerotic encephalopathy.

OBJECTIVES: To examine the CSF concentrations of molecules reflecting demyelination, neuronal and axonal degeneration, gliosis, monoaminergic neuronal function, and aminergic and peptidergic neurotransmission in a large series of patients with normal pressure hydrocephalus (NPH) or subcortical arteriosclerotic encephalopathy (SAE), to elucidate pathogenic, diagnostic, and prognostic features. METHODS: CSF concentrations of glycosphingolipid (sulfatide), proteins (neurofilament triplet protein (NFL), glial fibrillary acidic protein (GFAP)), neuropeptides (vasoactive intestinal peptide (VIP), 4-aminobutyric acid (GABA)), and monoamines (homovanillic acid (HVA), 5-hydroxy-indoleacetic acid (5-HIAA), 4-hydroxy-3-methoxyphenylglycol (HMPG)) were analysed in 43 patients with NPH and 19 patients with SAE. The diagnoses of NPH and SAE were based on strict criteria and patients with NPH were subsequently operated on. Twelve clinical variables, psychometric tests measuring perceptual speed, accuracy, learning, and memory and a psychiatric evaluation were performed in all patients and before and after a shunt operation in patients with NPH. RESULTS: The CSF sulfatide concentration was markedly increased in patients with SAE (mean 766, range 300-3800 nmol/l) compared with patients with NPH (mean 206, range 50-400 nmol/l) (p<0.001). 5-HIAA, GABA, and VIP in CSF were higher in patients with SAE than in patients with NPH. The patients with NPH with cerebrovascular aetiology had higher sulfatide concentrations and a poorer outcome after shunt surgery than patients with NPH with other aetiologies. CONCLUSIONS: The pathogenesis of the white matter changes in NPH and SAE is different and ischaemic white matter changes can be a part of the NPH state. The markedly increased CSF sulfatide concentrations in patients with SAE indicate ongoing demyelination as an important pathophysiological feature of SAE. The CSF sulfatide concentration distinguished between patients with SAE and those with NPH with a sensitivity of 74% and a specificity of 94%, making it an important diagnostic marker.

Adult↗

Tricyclic imipramine modification of the circadian rhythms of hypothalamic serotonin, its precursors and acid catabolite in individually housed rats.

Circadian rhythms of serotonin (5HT), its precursors tryptophan (TP) and 5-hydroxy-tryptophan (5HTP) and its acid catabolite 5-hydroxy-indoleacetic acid (5HIAA), were determined in the hypothalamus of control rats and rats which had been treated continuously with subcutaneous imipramine (10 mg/kg/day) for 2 weeks. Rats were individually housed and entrained to LD 12:12. Controls showed the 5HT and TP peaks in the light and dark periods respectively, as reported in the literature, but no inverted correlation (antiphase) between 5HT and 5HIAA rhythms. Imipramine significantly modified circadian rhythm characteristics: the 5HT acrophase was advanced, that of TP and 5HIAA was delayed. Imipramine also significantly increased hypothalamic 5HT and TP concentrations.

5-Hydroxytryptophan↗

[Changes in monoamine metabolites in the nucleus accumbens and locomotor activity by reserpine: an in vivo microdialysis study].

The effect of ip administration of reserpine (2 mg/kg) on the monoamine metabolites 3,4-dihydroxyphenyl acid (DOPAC), homovanillic acid (HVA), 5-hydroxy-indoleacetic acid (5-HIAA) was estimated in the freely moving rat by intracerebral dialysis in the nucleus accumbens. Locomotor activity was measured simultaneously by Automex. Locomotor activity tended to increase during the first 2 hours. On the other hand, a significant reduction of locomotor activity was found during the 3rd-5th hour period of the experiment. HVA-total (conjugated + free) and 5-HIAA were elevated significantly (P less than 0.05) during the 3rd-5th and the 5th-6th hour periods, respectively. However, the difference of DOPAC between both groups did not reach statistical significance throughout the experiment. During the 3rd-5th hour of experiment, the change of behavior and HVA were inversely correlated. This might suggest that an HVA level in the dialysate is a good index of dopamine metabolism in the nucleus accumbens after reserpine treatment.

3,4-Dihydroxyphenylacetic Acid↗

Analysis of monoamines in the cerebrospinal fluid of Chinese patients with neurological diseases.

Levels of dopamine (DA), 3-methoxy-4-hydroxy-phenylacetic acid (HVA), 5-hydroxy-indoleacetic acid (HIAA), 5-hydroxytryptamine (HT), 5-hydroxytryptophan (HTP), and 3, 4-dihydroxyphenylalanine (DOPA) in cerebrospinal fluids (CSF) of twenty-four Chinese with or without neurological diseases were determined simultaneously. The level of HIAA in the CSF of myelopathy-spondylosis patients and the level of DA in CSF of the myelopathy-tumor patients were higher than that of the control. No difference in levels of monoamines and their metabolites was observed between the control and patients with dementia.

Adolescent↗

[Late chronic dyskinesia from neuroleptic drugs (CSF and pharmacological data) (author's transl)].

CSF values of dopamine and serotonine metabolites (homovanillic acid and 5-hydroxy-indoleacetic acid) have been studied in 4 patients suffering from late dyskinesia due to neuroleptics. Trazodone (which acts on all monoaminergic systems, namely noradrenergic, dopaminergic and serotoninergic) induced a marked clinical improvement associated with CSF HVA increase and 5-HIAA decrease. The Authors suggest that involuntary movements could be due to an impairment of 5-HT e DA link.

Adult↗

Purification and characterization of the flavoprotein tryptophan 2-monooxygenase expressed at high levels in Escherichia coli.

Tryptophan 2-monooxygenase from Pseudomonas savastanoi is a flavoprotein which catalyzes the formation of indoleacetamide from tryptophan. This is the first step in a two-step pathway for the formation of indoleacetic acid during infection of plants and subsequent gall formation by this and other bacteria. The enzyme has been expressed in Escherichia coli at high levels, and a purification procedure has been developed which generates micromolar amounts of protein. The purified enzyme contains tightly bound indoleacetamide; a method involving dialysis against 20% methanol has been developed for removing the indoleacetamide without significant loss of enzyme activity. Amino acids with large hydrophobic side chains are the best substrates; N-substituted phenylalanines will also act as substrates. N-ethylmaleimide, methyl methanethiol-sulfonate, and diethylpyrocarbonate act as active site-directed reagents, consistent with a histidine and a cysteine at or near the enzyme active site. Vinylglycine partially inactivates the enzyme, while propargylglycine has no effect.

Amino Acids↗

Mechanistic studies of the flavoenzyme tryptophan 2-monooxygenase: deuterium and 15N kinetic isotope effects on alanine oxidation by an L-amino acid oxidase.

Tryptophan 2-monooxygenase (TMO) from Pseudomonas savastanoi catalyzes the oxidative decarboxylation of l-tryptophan during the biosynthesis of indoleacetic acid. Structurally and mechanistically, the enzyme is a member of the family of l-amino acid oxidases. Deuterium and 15N kinetic isotope effects were used to probe the chemical mechanism of l-alanine oxidation by TMO. The primary deuterium kinetic isotope effect was pH independent over the pH range 6.5-10, with an average value of 6.0 +/- 0.5, consistent with this being the intrinsic value. The deuterium isotope effect on the rate constant for flavin reduction by alanine was 6.3 +/- 0.9; no intermediate flavin species were observed during flavin reduction. The kcat/Kala value was 1.0145 +/- 0.0007 at pH 8. NMR analyses gave an equilibrium 15N isotope effect for deprotonation of the alanine amino group of 1.0233 +/- 0.0004, allowing calculation of the 15N isotope effect on the CH bond cleavage step of 0.9917 +/- 0.0006. The results are consistent with TMO oxidation of alanine occurring through a hydride transfer mechanism.

Alanine↗

Identification of indole-3-acetic acid producing freshwater wetland rhizosphere bacteria associated with Juncus effusus L.

Production of indole-3-acetic acid (IAA), a key physiological feature of culturable, O2-tolerant bacteria associated with the freshwater macrophyte Juncus effusus L., was examined over a period of 2 years. Up to 74% of rhizobacteria identified and tested produced IAA. The number of indoleacetic acid producers decreased in winter. IAA was produced even when L-tryptophan, a precursor of IAA, was not added to the medium. Most of the IAA-producing strains were dominated by strains that were not identifiable to species level on the basis of API testing. Based on 16S rRNA gene sequencing and fatty acid analysis, it was found that IAA-producing rhizosphere bacteria associated with the freshwater wetland plant Juncus effusus L. are representatives of several families, including the Enterobacteriaceae, Pseudomonadaceae, Aeromonadaceae, Burkholderiaceae, and Bacillaceae. This study identifies numerous potentially important bacterial physiological groups of freshwater wetlands. Additionally, the study provides a baseline for monitoring and assessing the mutualistic relationships of wetland plants with rhizosphere bacteria in freshwater wetlands.

Aeromonadaceae↗

Altered growth response to exogenous auxin and gibberellic acid by gravistimulation in pulvini of Avena sativa.

Pulvini of excised segments from oats (Avena sativa L. cv Victory) were treated unilaterally with indoleacetic acid (IAA) or gibberellic acid (GA3) with or without gravistimulation to assess the effect of gravistimulation on hormone action. Optimum pulvinus elongation growth (millimeters) and segment curvature (degrees) over 24 hours were produced by 100 micromolar IAA in vertical segments. The curvature response to IAA at levels greater than 100 micromolar, applied to the lower sides of gravistimulated (90 degrees) pulvini, was significantly less than the response to identical levels in vertical segments. Furthermore, the bending response of pulvini to 100 micromolar IAA did not vary significantly over a range of presentation angles between 0 and 90 degrees. In contrast, the response to IAA at levels less than 10 micromolar, with gravistimulation, was approximately the sum of the responses to gravistimulation alone and to IAA without gravistimulation. This was observed over a range of presentation angles. Also, GA3 (0.3-30 micromolar) applied to the lower sides of horizontal segments significantly enhanced pulvinus growth and segment curvature, although exogenous GA3 over a range of concentrations had no effect on pulvinus elongation growth or segment curvature in vertical segments. The response to GA3 (10 micromolar) plus IAA (1.0 or 100 micromolar) was additive for either vertical or horizontal segments. These results indicate that gravistimulation produces changes in pulvinus responsiveness to both IAA and GA3 and that the changes are unique for each growth regulator. It is suggested that the changes in responsiveness may result from processes at the cellular level other than changes in hormonal sensitivity.

Avena↗