AIMTo establish a suitable condition for extraction of phenylalanine (Phe), 5-hydroxytryptophan (5-OH-Trp) and four diastereomeric salts, (1R,2S)-ephedrine- (2S,3S)-tartaric acid, (1R,2S)-ephedrine-(2R,3R)-tartaric acid, (1S,2S)-pseudoephedrine-(2S,3S)-tartaric acid, (1S,2S)-pseudoephedrine- (2R,3R)-tartaric acid in supercritical fluid extraction and to assess the solubilities of Phe, 5-OH-Trp and the four diastereomeric salts in CO2.

METHODSSingle-pass method and HPCE.

RESULTSThe solubilities of Phe, 5-OH-Trp and the four diastereomeric salts in CO2 were determined over temperature and pressure ranges of 25-50 degrees C and 6.32-34.03 MPa respectively. The experimental results showed that the solubilities of Phe, 5-OH-Trp and the four diastereomeric salts do not increase with density of CO2. There existed a maximum in the critical region of CO2.

CONCLUSIONThe dramatically high solubilities in the pressure of 6.32-7.78 MPa show a critical behavior, which can be explained by critical characteristic through thermodynamics analysis. The results suggest that the separation of Phe, 5-OH-Trp and the four diastereomeric salts is more efficient in critical region of CO2.

5-Hydroxytryptophan ; chemistry ; Ephedrine ; chemistry ; Phenylalanine ; chemistry ; Pressure ; Solubility ; Stereoisomerism ; Tartrates ; chemistry ; Temperature

In order to demonstrate autoradiographically the sites of serotonin metabolism in the brain, DL 5-HTP and DL 5-HTP-C14 were intraperitoneally administered to healthy adult mice. In order to distinguish histochemically serotonin-like substances which have staining characteristics similar to the enterochromaffin cells of the gastrointestinal tract, serotonin-releaser reserpine was administered intravenously to healthy adult rabbits. A stripping film technique for autoradiography, a ferric ferricyanide reduction test by Schmorl, and a Gomori-Burtner methenamine silver staining method for argentaffin cells were used in this study. In the brain tissues of mice treated with 5-HTP, it was observed that the cytoplasm of the nerve cells, of the cerebral cortex had blue positive staining substances by the ferric ferricyanide technique. In similar tissue sites in mice treated with 5-HTP-C14, a number of blackened-par-ticles reduced by beta rays were easily found. especially in the cytoplasm of nerve cells and neuroglia cells. It is suggested that the serotonin precursor, DL 5-hydrox-ytryptophan is metabolized the cerebral tissue, and serotonin is synthesized also in the nerve cells and the neuroglia cells.
5-Hydroxytryptophan/diagnostic use ; Animals ; Brain/*metabolism ; Injections, Intraperitoneal ; Injections, Intravenous ; Mice ; Reserpine/diagnostic use ; Serotonin/*metabolism ; Staining and Labeling

To investigate the distribution, ultrastructure and synapsis of serotoninergic cells and CGRP nerve fibers in mammalian taste buds, immunohistochemistry and electronmicroscopy were applied to mice vallate papillae. In normal mice, 1~2 serotonin immunoreactive cells were present in each taste bud section. After preloading 5-HTP, 3~6 cells showed strong immunoreactivity for serotonin. These cells were elongated, and their cytoplasm extended from the taste pore to the base of the taste bud. CGRP nerve fibers formed thick subgemmal nerve plexus under the basal lamina, and branched varicose perigemmal and intragemmal nerve fibers. Under the electron-microscope, three types of taste cells; dark cell, light cell and basal cells, were identified by their shape, location and electrical densities. Immuno-electronmicroscopy revealed that serotoninergic cells were dark cells. CGRP nerve fibers were located in and around taste buds, but the synaptic contacts with taste cells was not found. These findings indicate that mice taste cells are consisted of dark cell, light cell and basal cells, and dark cells contain serotonin. And, CGRP nerve fibers in taste buds may function as general sensory fibers.
5-Hydroxytryptophan ; Animals ; Basement Membrane ; Calcitonin Gene-Related Peptide* ; Calcitonin* ; Chromosome Pairing ; Cytoplasm ; Immunohistochemistry ; Mice* ; Nerve Fibers* ; Serotonin ; Taste Buds*

Tetrahydrobiopterin (BH4) deficiency is caused by mutations in genes encoding enzymes involved in the synthesis and regeneration of BH4. The condition is usually accompanied by hyperphenylalaninemia (HPA) and deficiency of neurotransmitter precursors L-dopa and 5-hydroxytryptophan. BH4 deficiency is much rarer than classical phenylketonuria. Dihydropteridine reductase (DHPR) deficiency, an autosomal recessive genetic disorder, is a cause of malignant hyperphenylalaninemia due to BH4 deficiency. When left untreated, DHPR deficiency leads to neurologic deterioration at the age of 4 or 5 months, including psychomotor retardation, tonicity disorders, drowsiness, irritability, abnormal movements, hyperthermia, hypersalivation, and difficulty swallowing. Treatment of DHPR deficiency should be initiated as early as possible with BH4 supplementation and replacement of the neurotransmitter precursors L-dopa and 5-hydroxytryptophan. We report the first case of DHPR deficiency in Korea, a child diagnosed at 9 years of age by genetic testing.
5-Hydroxytryptophan ; Biopterin ; Child ; Deglutition ; Dihydropteridine Reductase ; Dyskinesias ; Fever ; Genetic Testing ; Humans ; Korea ; Levodopa ; Neurotransmitter Agents ; Phenylketonurias ; Regeneration ; Sialorrhea ; Sleep Stages

This study was conducted to define the underlying mechanism of hypophagia induced by increased central serotonergic action. Rats received 3 daily injections of 5-hydroxy-L-tryptophan (5-HTP), a serotonin precursor, at a dose of 100 mg/kg/10 ml saline at 1 h before lights off. A significant suppression in food intake was observed shortly after the 5-HTP injection and persisted during 3 daily 5-HTP injections. Neuropeptide Y (NPY) expression in the arcuate nucleus increased after 3 days of 5-HTP treatment, as high as in the pair-fed group. Immunoreactivity of phosphorylated extracellular signal-regulated protein kinase (pERK1/2) in the hypothalamic paraventricular nucleus (PVN) was increased markedly by 3 days of 5-HTP treatment, but not by 3 days of pair-fed. mRNA expression levels of serotonin reuptake transporter (5-HTT) was increased in the dorsal raphe nucleus of the 5-HTP treated rats, but not in the pair-fed group. Results suggest that increased pERK1/2 in the PVN of 5-HTP injected rats may be a part of serotonergic anorectic signaling, perhaps blunting the orectic action of NPY; i.e., 5-HTP injected rats showed hypophagia despite of increased NPY expression in the arcuate nucleus.
5-Hydroxytryptophan ; Animals ; Arcuate Nucleus ; Eating ; Hypothalamus ; Light ; Neuropeptide Y ; Paraventricular Hypothalamic Nucleus ; Protein Kinases ; Raphe Nuclei ; Rats ; RNA, Messenger ; Serotonin

Hyperphenylalaninemia can result from defects in either the phenylalanine hydroxylase (PAH) enzyme or in the synthesis or recycling of the active pterin, tetrahydrobiopterin (BH4), which is an obligate co-factor for the PAH enzyme, as well as tyrosine hydroxylase and tryptophan hydroxylase. One of the most common causes of BH4 deficiency is a defect in the synthesis of 6-pyruvoyltetrahydropterin synthase (PTPS) enzyme. Patients present with progressive neurological disease such as mental retardation, convulsions and disturbance of tone and posture despite strict adherence to diet and good metabolic control. The authors report the first two cases of PTPS deficiency in the Philippines. Both are females with initial phenylalanine levels of more than 1300 umol/L who continued to develop neurologic deterioration despite good metabolic control and strict adherence to diet. Further investigation showed that they both had PTPS deficiency. Treatment was started with BH4, L-dopa/carbidopa, and 5-hydroxytryptophan (5HT) with concomitant significant improvements in their neurologic and developmental outcomes.

Human ; Female ; Child Preschool ; Infant ; Phenylalanine Hydroxylase ; Carbidopa ; Tyrosine 3-monooxygenase ; 5-hydroxytryptophan ; Tryptophan Hydroxylase ; Levodopa ; Sapropterin ; Intellectual Disability ; Philippines ; Phenylketonurias ; Pterins ; Seizures ; Diet ; Posture

OBJECTIVETo emphasize early differential diagnosis from patients with hyperphenylalaninemia (HPA) and to evaluate the treatment and long-term outcome of patients with tetrahydrobiopterin synthase (BH4) deficiency in Northern Chinese population.

METHODSFrom 1992 to 2005, a total of 618 patients with HPA were diagnosed and/or cared for in our outpatient clinic. Urinary pterin analysis, detection of dihydropteridine reductase (DHPR) activity in blood, and then BH4 loading tests were carried out to differentiate BH4 deficiency in these patients from classical phenylketonuria. BH4 deficient patients were treated with BH4, levodopa and 5-hydroxytryptophane (5-HTP) immediately while the diagnosis was done to disease. Patientso blood phenylalanine levels, psychomotor and intelligence development were followed up.

RESULTSA total of 38 cases were diagnosed as BH4 deficiency, all of them were revealed as 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency from the extremely decreased urine biopterin, normal DHPR activities and drop down of blood phenylalanine level to normal range within 4 to 8 hours after BH4 loading. The most common manifestations were progressively psychomotor and mental retardation to patients even after taking early dietary treatment. The patients were diagnosed and treated with drugs at the ages of 2.1 months to 13 years. With 4 patients died of pneumonia, 7 patients refused to treatment, only 27 patients were under treatment and followed up. The average full scale development or intelligence quotient (DQ/IQ) of patients who were treated within and after 6 months were 86+/- 10 or 66+/- 7 respectively. Development was not even in different aspects. A significant negative correlation was observed between the level of the DQ and the age of treatment commenced (r was -0.714, P< 0.01). Eleven patients experienced the extrapyramidal movement disorders, 3 of them combined with epilepsy. The extrapyramidal disorders were controlled by administration of levodopa.

CONCLUSIONThe differential diagnosis for BH4 deficiency should be carried out in all patients with HPA. PTPS deficiency is the most common form of BH4 deficiency in Northern Chinese population. The long-term outcome of these patients benefits from diagnosis and treatment with BH4, levodopa and 5-HTP as early as possible.

5-Hydroxytryptophan ; therapeutic use ; Asian Continental Ancestry Group ; genetics ; Biopterin ; analogs & derivatives ; deficiency ; therapeutic use ; Child, Preschool ; China ; Dihydropteridine Reductase ; blood ; Humans ; Infant ; Levodopa ; therapeutic use ; Phenylalanine ; blood ; Phenylketonurias ; drug therapy ; genetics ; metabolism ; Phosphorus-Oxygen Lyases ; deficiency ; genetics

BACKGROUND/OBJECTIVES: The aim of this study was to evaluate the biological and sleep-promoting effects of combined γ-aminobutyric acid (GABA) and 5-hydroxytryptophan (5-HTP) using caffeine-induced sleepless fruit flies, ICR mice, and Sprague-Dawley rats. MATERIALS/METHODS: Video-tracking analysis was applied to investigate behavioral changes of Drosophila melanogaster. Pentobarbital-induced sleep test and electroencephalogram (EEG) patterns were used for analysis of sleep latency, duration, and quantity and quality of sleep in vertebrate models. RESULTS: Administration of combined GABA/5-HTP could significantly reverse the caffeine induced total distance of flies (P < 0.001). Also, individually administered and combined GABA/5-HTP significantly increased the total sleeping time in the caffeine-induced sleepless ICR mice (P < 0.001). In the caffeine-induced sleepless SD-rats, combined GABA/5-HTP showed significant differences in sleep quality between individual amino acid administrations (P < 0.05). CONCLUSIONS: Taken together, we identified inhibitory effects of combined GABA/5-HTP in locomotor activity, sleep quantity and quality in caffeine-induced sleepless models, indicating that combined GABA/5-HTP may be effective in patients with insomnia by providing sufficient sleep.
5-Hydroxytryptophan ; Amino Acids* ; Animals ; Caffeine ; Diptera ; Drosophila melanogaster ; Electroencephalography ; Fruit ; gamma-Aminobutyric Acid ; Humans ; Mice ; Mice, Inbred ICR ; Motor Activity ; Rats, Sprague-Dawley ; Sleep Initiation and Maintenance Disorders ; Vertebrates