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PubMed2004

Evaluation of two nonisotopic immunoassays for determination of glutamic acid decarboxylase and tyrosine phosphatase autoantibodies in serum.

BACKGROUND: Autoantibodies for the 65-kDa form of glutamic acid decarboxylase (GAD65) and protein tyrosine phosphatase-like protein (IA-2) are measured for risk prediction and diagnosis of autoimmune diabetes mellitus. There is a lack of adequate nonisotopic alternatives to the most widely used method for both autoantibodies, which is a radiobinding assay (RBA). METHODS: We compared two commercially available immunoassays, an ELISA and a time-resolved immunofluorometric assay (TR-IFMA), with RBA. RESULTS: We found excellent agreement between the RBA and ELISA for measurement of GAD65 autoantibodies (GADAs); they showed comparable analytical precision in the cutoff range and achieved similar diagnostic specificity. The ELISA identified more GADA-positive individuals among patients with new-onset type 1 diabetes than did the RBA [89% (95% confidence interval, 78-95%), vs 71% (58-82%); P <0.03]. For IA-2 autoantibodies (IA-2As), only the TR-IFMA achieved analytical performance and diagnostic accuracy comparable to that of the RBA. These results with the GADA ELISA and IA-2A TR-IFMA were consistent with those obtained blindly in the Diabetes Antibody Standardization Program 2003. The performance of the GADA TR-IFMA and IA-2A ELISA was unsatisfactory, and these tests were not subjected to clinical evaluation. CONCLUSIONS: The GADA ELISA and IA-2A TR-IFMA behave comparably with RBA and are thus suitable for use in the clinical laboratory.

Adolescent↗
PubMed2003

Localization and functional significance of striatal neurons immunoreactive to aromatic L-amino acid decarboxylase or tyrosine hydroxylase in rat Parkinsonian models.

Striatal neurons which are immunoreactive (ir) to aromatic L-amino-acid decarboxylase (AADC) or tyrosine hydrodroxylase (TH) may play a role in the decarboxylation of L-DOPA to dopamine (DA) in advanced stages of Parkinson's disease (PD). However, the functional significance of these neurons and the mechanisms responsible for their induction remain to be clarified. In this study, rats were subjected to different types of dopaminergic or serotonergic denervation and L-DOPA injection to study the effects on these neurons. AADC-ir neurons were found in both normal and DA-denervated striata, and no significant differences in their number and distribution were induced following different types of denervation or L-DOPA administration. TH-ir neurons were only found in DA-denervated striata. However, TH-ir neurons did not appear in those areas with maximal DA depletion, but rather were observed near spared or partially lesioned DA terminals. The population of AADC-ir neurons may make a significant contribution to the effects of exogenous L-DOPA in advanced stages of PD. In addition, TH-ir neurons may contribute to these effects, since we have detected AADC-ir in TH-ir neurons using confocal laser scanning microscopy. Finally, neither L-DOPA therapy nor serotonergic denervation induces significant changes in the number or distribution of these neurons.

Animals↗
PubMed1991

Differential activation of ornithine decarboxylase and tyrosine kinase in the rectal mucosa of patients with hyperplastic and adenomatous polyps.

Hyperplastic polyps are considered to be benign colonic lesions with almost no potential for malignant transformation. Recent reports have shown an increased association of hyperplastic polyps with adenomatous polyps and have advocated a full colonoscopy in patients who harbor hyperplastic polyps. Hyperproliferative mucosa is known to be associated with adenomatous polyps, but its relationship to hyperplastic polyps is unknown. In the present pilot study, it is determined whether a change in mucosal proliferative patterns is observed in patients who harbor only hyperplastic polyps or a history of hyperplastic polyps relative to those who harbor both hyperplastic polyps and adenomatous polyps by measuring ornithine decarboxylase and tyrosine kinase activity in macroscopically normal rectal mucosa. Fifteen patients had either adenomatous polyps proximally or harbored adenomatous polyps and hyperplastic polyps. Seven patients had hyperplastic polyps and 15 patients had a prior history of hyperplastic polyps with no polyps found during the current examination. The ornithine decarboxylase activity of the rectal mucosa with proximal adenomatous polyps or both polyp types was significantly higher than that of hyperplastic polyps, the history of hyperplastic polyps, or controls, and values for hyperplastic polyps and the history of hyperplastic polyps were similar to controls. On the other hand, tyrosine kinase activity in the rectal mucosa of patients with both or either polyp type was elevated without any significant difference between hyperplastic and adenomatous polyps. Thus, it is concluded that although increased ornithine decarboxylase activity in rectal mucosa suggests the presence of adenomatous polyps or a combination of adenomatous with hyperplastic polyps, increased tyrosine kinase activity suggests the presence of any type of polyp.

Colonic Polyps↗
PubMed1995

Striatal dihydroxyphenylalanine decarboxylase and tyrosine hydroxylase protein in idiopathic Parkinson's disease and dominantly inherited olivopontocerebellar atrophy.

We measured the levels of dopamine, tyrosine hydroxylase (TH) protein, and dihydroxyphenylalanine (DOPA) decarboxylase (DDC) protein in the striatum of 10 patients with idiopathic Parkinson's disease (PD) and 23 patients with dominantly inherited olivopontocerebellar atrophy (OPCA). The levels of dopamine were markedly reduced (2% of control) in the striatum of the patients with PD, whereas striatal dopamine in the patients with OPCA ranged from normal (> 60% of control) to moderately reduced (20-60% of control) to severely depleted (< 20% of control). Both TH and DDC protein levels were significantly lower than those of the controls in the striatum of all of the patients with PD and in the subgroup of patients with OPCA having severely depleted dopamine. In contradistinction, TH but not DDC protein levels were reduced in those patients with OPCA having moderately reduced dopamine levels. This suggests that in the early stage of nigrostriatal dopamine neurone degeneration, DDC levels may be less susceptible to neurodegenerative influences than is TH synthesis or, alternatively, DDC synthesis may be more aggressively upregulated. Unexpectedly, from the blot immunolabeling analysis an additional DDC-immunoreactive band of slightly lower apparent molecular mass was detected in two of the patients with PD and in 12 of the patients with OPCA. This additional DDC band, which was not present in any of the control subjects, may reflect posttranslational modification(s) of DDC related to the neurodegenerative process.

Aged↗
PubMed1995

Ornithine decarboxylase and tyrosine kinase activity in juvenile polyps of childhood.

Juvenile polyps (JP) are the most common colonic tumor in children. Although considered benign, malignant transformation has been reported in JP. Ornithine decarboxylase (ODC) and tyrosine kinase (TyK) enzymes are markers for a rapid cell proliferation index. DNA aneuploidy score and p53 gene expression are late malignant changes seen in patients with colon cancer. In this study, we investigated ODC and TyK activities as well as DNA aneuploidy score and p53 expression in juvenile polyps compared with the adjacent normal colonic mucosa. Results showed that ODC was significantly increased in JP compared with the adjacent normal colonic mucosa. TyK activity was increased in 3/5 polyps and decreased in 2/5 polyps compared with the mucosa. Mean TyK activity was higher in JP compared with normal mucosa but did not reach significance (707 and 632 pmol/mg pmol, respectively). Moreover, changes in phosphorylization of TyK proteins was also observed in JP but not in normal mucosa. JP had a normal DNA aneuploidy score and showed no expression of p53 gene. We conclude that JP do not express p53 gene and aneuploidy but had higher activity of ODC and TyK enzymes, suggesting a higher stage of cell proliferation.

Adolescent↗
PubMed1999

Expression of cholecystokinin, somatostatin, thyrotropin-releasing hormone, glutamic acid decarboxylase and tyrosine hydroxylase genes in the central nervous motor systems of the genetically dystonic hamster.

In the dt(sz) mutant hamster with idiopathic generalized dystonia, functional abnormalities of several neurotransmitters have been suggested to play a role in the development of symptoms. In the present study, we have used histochemistry with (35)S-ATP labeled oligonucleotides to determine whether these abnormalities are associated with modulation in the expression of neurotransmitter genes in motor regions. We examined the expression of genes encoding cholecystokinin (CCK), somatostatin (SRIF), thyrotropin-releasing hormone (TRH), glutamic acid decarboxylase (GAD), tyrosine hydroxylase (TH) and growth-associated protein 43 (GAP43) in the cortex and basal ganglia of dystonic hamsters and of non-dystonic control hamsters of a related inbred line and of a non-related outbred line. The distribution of these mRNAs in normal hamster brain was similar to that in normal rat brain. In all cortical regions studied (frontal, parietal and piriformis), the expression of CCK was similar in dystonic and inbred controls but was significantly greater than in outbred controls. In the anterior thalamus, CCK expression was lower in dystonic hamsters than in both control groups. SRIF expression was significantly decreased in the cortex and striatum of dystonic animals than in inbred and outbred control hamsters. GAD expression was lower in the striatum and substantia nigra, pars reticulata of dystonic than in outbred hamsters, but similar values were found in all groups in the other regions studied. TH was lower in the substantia nigra of dystonic than in inbred controls. No changes were found in GAP43 expression. This study demonstrates that changes in modulation of the expression of some peptides and neurotransmitter enzymes can be found in the dystonic hamster, which is in contrast to other animal models such as the dystonic rat, where no such changes have been found. The present data are consistent with previous findings in dt(sz) hamsters that suggest a dysfunction within the basal ganglia-thalamocortical circuits.

Animals↗
PubMed1993

Changes in gastric mucosal ornithine decarboxylase and tyrosine phosphorylation of proteins in postgastrectomy patients.

There is evidence to suggest that partial gastrectomy is associated with an increased risk of developing gastric carcinoma in humans. Since ornithine decarboxylase (ODC) and tyrosine phosphorylation of proteins are involved in the regulation of cell proliferation, the present study was undertaken to examine the time-dependent changes of these variables in the postgastrectomy stomach. Thirty-seven postgastrectomy patients (Billroth I (BI), n = 7, and Billroth II (BII), n = 30) underwent gastroscopy. For comparison, five patients with intact stomachs (three healthy and two postvagotomy and pyloroplasty) were also studied. Gastric mucosal biopsy specimens were obtained within 5 cm of the anastomosis and analyzed for ODC activity. In addition, tyrosine phosphorylation of membrane proteins was also determined in representative samples of BII patients. Gastric mucosal ODC activity was significantly higher in BII patients in whom gastrectomy had been performed > 15 years earlier compared with those in whom it had been performed < 15 years earlier (p < 0.001) or controls (p = 0.004). Although the mean ODC activity was higher in BII than in BI patients, the difference was not significant (p = 0.103). Isolated patients with high ODC activity demonstrated increased phosphorylation of tyrosine membrane proteins with M(r) of 55-60.

Adult↗
PubMed1985

Coexistence of immunoreactivities for glutamate decarboxylase and tyrosine hydroxylase in some neurons in the periglomerular region of the rat main olfactory bulb: possible coexistence of gamma-aminobutyric acid (GABA) and dopamine.

The coexistence of immunoreactivities for glutamate decarboxylase (GAD) and tyrosine hydroxylase (TH) was revealed in some neurons in the periglomerular region and in the superficial part of the external plexiform layer of the rat main olfactory bulb. In neurons showing the immunoreactivity for either GAD or TH, about 10-55% showed both immunoreactivities.

Animals↗
PubMed2006

The mechanism of the tyrosine transporter TyrP supports a proton motive tyrosine decarboxylation pathway in Lactobacillus brevis.

The tyrosine decarboxylase operon of Lactobacillus brevis IOEB9809 contains, adjacent to the tyrosine decarboxylase gene, a gene for TyrP, a putative tyrosine transporter. The two genes potentially form a proton motive tyrosine decarboxylation pathway. The putative tyrosine transporter gene of L. brevis was expressed in Lactococcus lactis and functionally characterized using right-side-out membranes. The transporter very efficiently catalyzes homologous tyrosine-tyrosine exchange and heterologous exchange between tyrosine and its decarboxylation product tyramine. Tyrosine-tyramine exchange was shown to be electrogenic. In addition to the exchange mode, the transporter catalyzes tyrosine uniport but at a much lower rate. Analysis of the substrate specificity of the transporter by use of a set of 19 different tyrosine substrate analogues showed that the main interactions between the protein and the substrates involve the amino group and the phenyl ring with the para hydroxyl group. The carboxylate group that is removed in the decarboxylation reaction does not seem to contribute to the affinity of the protein for the substrates significantly. The properties of the TyrP protein are those typical for precursor-product exchangers that operate in proton motive decarboxylation pathways. It is proposed that tyrosine decarboxylation in L. brevis results in proton motive force generation by an indirect proton pumping mechanism.

Amino Acid Transport Systems↗
PubMed1999

The trace amine tyramine is essential for sensitization to cocaine in Drosophila.

BACKGROUND: Sensitization to psychostimulant drugs of abuse is thought to be an important aspect of human addiction, yet how it develops is still unclear. The development of sensitization to cocaine in the fruit fly Drosophila melanogaster is strikingly similar to that observed in vertebrates. By taking advantage of the powerful genetic approaches that are possible in Drosophila, we are able to identify and characterize mutants that fail to develop sensitization. RESULTS: We found that the Drosophila mutant inactive (iav) failed to become sensitized to cocaine. Mutant flies had reduced amounts of the trace amine tyramine in the brain because of reduced activity of the enzyme tyrosine decarboxylase (TDC), which converts tyrosine to tyramine. Furthermore, cocaine exposure induced TDC enzyme activity in a time-dependent manner that paralleled the development of behavioral sensitization. The sensitization failure of iav flies could be rescued by feeding the flies with tyramine; other biogenic amines or amine precursors did not have the same effect. CONCLUSIONS: These results indicate an essential role for tyramine in cocaine sensitization in Drosophila.

Animals↗
PubMed2005

Ionization state of pyridoxal 5'-phosphate in D-serine dehydratase, dialkylglycine decarboxylase and tyrosine phenol-lyase and the influence of monovalent cations as inferred by 31P NMR spectroscopy.

The 31P NMR spectroscopy of three pyridoxal 5'-phosphate-dependent enzymes, monomeric D-serine dehydratase, tetrameric dialkylglycine decarboxylase and tetrameric tyrosine phenol-lyase, whose enzymatic activities are dependent on alkali metal ions, was studied. 31P NMR spectra of the latter two enzymes have never been reported, their 3D-structures, however, are available. The cofactor phosphate chemical shift of all three enzymes changes by approximately 3 ppm as a function of pH, indicating that the phosphate group changes from being monoanionic at low pH to dianionic at high pH. The 31P NMR signal of the phosphate group of pyridoxal 5'-phosphate provides a measure of the active site changes that occur when various alkali metal ions are bound. Structural information is used to assist in the interpretation of the chemical shift changes observed. For D-serine dehydratase, no structural data are available but nevertheless the metal ion arrangement in the PLP binding site can be predicted from 31P NMR data.

Carboxy-Lyases↗
PubMed1978

[Aromatic amino acid metabolism in Enterobacteriaceae microorganisms].

The authors revealed phenylalanine deaminase (PAD) in the majority of the Citrobacter strains investigated; the activity of PAD varied within a rather considerable range (0.3--4.58 micrometer of phenylpiruvate in 1 hr per 1 mg of bacterial protein). The presence of this enzyme thus served as an auxiliary biochemical test characterising this group of conditionally pathogenic microbes of the Enterobacteriacea family. Tyrosine decarboxylase was absent in 26 of 50 strains of Citrobacter examined. In the rest of the strains the activity of this enzyme was low. Consequently, tyrosine decarboxylase could not be used for identification of microorganisms of the Citrobacter genus.

Amino Acid Oxidoreductases↗
PubMed1999

Requirement of circadian genes for cocaine sensitization in Drosophila.

The circadian clock consists of a feedback loop in which clock genes are rhythmically expressed, giving rise to cycling levels of RNA and proteins. Four of the five circadian genes identified to date influence responsiveness to freebase cocaine in the fruit fly, Drosophila melanogaster. Sensitization to repeated cocaine exposures, a phenomenon also seen in humans and animal models and associated with enhanced drug craving, is eliminated in flies mutant for period, clock, cycle, and doubletime, but not in flies lacking the gene timeless. Flies that do not sensitize owing to lack of these genes do not show the induction of tyrosine decarboxylase normally seen after cocaine exposure. These findings indicate unexpected roles for these genes in regulating cocaine sensitization and indicate that they function as regulators of tyrosine decarboxylase.

ARNTL Transcription Factors↗
PubMed1985

Application of avidin-ferritin and peroxidase as contrasting electron-dense markers for simultaneous electron microscopic immunocytochemical labelling of glutamic acid decarboxylase and tyrosine hydroxylase in the rat arcuate nucleus.

A pre-embedding immunostaining procedure was developed using ferritin and peroxidase to enable simultaneous electron microscopic localization of two antigens in the same tissue section. This method was used to study the anatomic relationship between glutamic acid decarboxylase (GAD) immunoreactive axons and tyrosine hydroxylase (TH) - containing neurons of the rat arcuate nucleus. The findings provide ultrastructural evidence that GAD-immunoreactive terminals establish symmetric (Gray II) synapses on TH-reactive neurons.

Animals↗
PubMed1989

Attenuation of azoxymethane-induced colonic mucosal ornithine decarboxylase and tyrosine kinase activity by calcium in rats.

Two in vivo and one in vitro studies were performed to evaluate the chemoprotective role of calcium during the early period of azoxymethane (AOM) induction. In the first set of experiments, groups of male Fischer 344 rats were s.c. injected with either AOM (20 mg/kg) or water (controls) and sacrificed immediately (0 time), and 1, 3, 5, and 7 days postinjection. In the second set of experiments, animals were injected with the same dose of AOM and subsequently pair-fed with rat chow containing either calcium carbonate or diet devoid of added calcium. The amount of calcium consumed was calculated to be 250 mg/kg b.w. In both experiments, colonic mucosa was assayed for ornithine decarboxylase (ODC). In addition, tyrosine kinase (Tyr-k) activity as well as tyrosine specific phosphorylation of membrane proteins were determined. Results revealed that maximal stimulation by AOM of ODC and Tyr-k activity occurred 5 days postinjection. This stimulation was significantly suppressed by calcium. AOM also produced an increase in the rate of tyrosine specific phosphorylation of two distinct colonic mucosal membrane proteins with Mr of 57,000 and 59,000. Again, dietary calcium suppressed the stimulation. In the third set of experiments, organ culture was utilized. Methylazoxymethanol, the active metabolite of AOM, was used instead of AOM in this part of the study. Four hour exposure of mucosal explants to methylazoxymethanol (1 microgram/ml) resulted in a significant (20-30%) increase in ODC and Tyr-k activity when compared to controls. Addition of either CaCl2 (2 mumol/ml) or difluoromethylornithine (2 nmol/ml) the irreversible inhibitor of ODC, significantly suppressed the methylazoxymethanol-induced activity of both ODC and Tyr-k. We conclude that calcium may have a chemoprotective role and tyrosine kinases may have a regulatory role in the early stages of AOM induction of colon cancer.

Animals↗
PubMed2004

Complex evolution of orthologous and paralogous decarboxylase genes.

The decarboxylases are involved in neurotransmitter synthesis in animals, and in pathways of secondary metabolism in plants. Different decarboxylase proteins are characterized for their different substrate specificities, but are encoded by homologous genes. We study, within a maximum-likelihood framework, the evolutionary relationships among dopa decarboxylase (Ddc), histidine decarboxylase (Hdc) and alpha-methyldopa hypersensitive (amd) in animals, and tryptophan decarboxylase (Wdc) and tyrosine decarboxylase (Ydc) in plants. The evolutionary rates are heterogeneous. There are differences between paralogous genes in the same lineages: 4.13 x 10(-10) nucleotide substitutions per site per year in mammalian Ddc vs. 1.95 in Hdc; between orthologous genes in different lineages, 7.62 in dipteran Ddc vs. 4.13 in mammalian Ddc; and very large temporal variations in some lineages, from 3.7 up to 54.9 in the Drosophila Ddc lineage. Our results are inconsistent with the molecular clock hypothesis.

Amino Acid Sequence↗
PubMed1992

Gamma-aminobutyric acid, glutamic acid decarboxylase and tyrosine hydroxylase in rat striatum demonstrated by single and dual immunocytochemistry.

By means of dual ultrastructural immunostaining the followings patterns are visualized: gamma-aminobutyric acid (GABA) immunoreactive neurons, dendrites, axons and axon terminals and tyrosine hydroxylase (TH) immunopositive fibers, varicosities and boutons in rat striatum. Additionally single glutamic acid decarboxylase (GAD) immunolabeling is carried out. Four subgroups of GABA and GAD immunoreactive striatal neurons are revealed. These neuronal types are identified on the basis of sectional diameters, nuclear form and nuclear envelope invaginations, quantity of cytoplasm and cell organelles. Plasmalemmal appositions between GABAergic and between GABAergic and immunonegative neurons are observed. All subgroups of striatal GABAergic neurons contact with GABA and GAD immunoreactive, TH immunoreactive and immunonegative boutons. In the striatal neuropil numerous GABAergic, dopaminergic and immunonegative axonal endings synapsed with dendrites and spines are found out. Massive dopaminergic striatal structures using dual immunostaining is evident. Some GABA and GAD immunoreactive dendrites are revealed in direct contact with capillary walls.

Animals↗