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Biomedical subjects

R F Butterworth

Publications and source records attributed to R F Butterworth.

At least 91 records · Page 5Linked to original sources
PubMed1996

Neuronal cell death in Wernicke's encephalopathy: pathophysiologic mechanisms and implications for PET imaging.

Thiamine deficiency in humans is associated with Wernicke's encephalopathy (WE) which is characterized neuropathologically by neuronal loss in selective brain regions. Pyrithiamine-induced thiamine-deficiency in the rat results in lesions which are similar in nature and distribution to those seen in human WE. Several mechanisms have been implicated in the pathogenesis of neuronal loss in thiamine deficiency including, (i) impaired cerebral energy metabolism, (ii) focal lactic acidosis, (iii) NMDA-receptor mediated excitotoxicity and (iv) blood-brain barrier breakdown. WE is difficult to diagnose during life and a large number of cases are missed by routine clinical neurological evaluation. Recently, non-invasive diagnostic procedures such as CT and MRI have been used for the evaluation of acute and chronic WE. Autoradiographic studies reveal that increased densities of binding sites for the astrocytic ligand 3H-PK11195 closely parallel the topographic distribution of reactive gliosis and neuronal loss in selective brain regions of pyrithiamine-induced thiamine-deficient rats. In contrast, binding sites for the neuronal ligand 3H-Ro15-1788 show poor regional correlation with neuronal loss in thiamine deficiency. Both of these ligands are available, and have been used in PET assessment of various disorders in humans. The results of autoradiographic studies suggest that 11C-PK11195 may offer a useful PET ligand for the assessment of brain damage in WE in humans.

Animals↗
PubMed1996

Alterations of thiamine phosphorylation and of thiamine-dependent enzymes in Alzheimer's disease.

There is a growing body of evidence to suggest that thiamine neurochemistry is disrupted in Alzheimer's Disease (AD). Studies in autopsied brain tissue from neuropathologically proven AD patients reveal significantly reduced activities of the thiamine phosphate dephosphorylating enzymes thiamine diphosphatase (TDPase) and thiamine monophosphatase (TMPase) as well as the thiamine diphosphate-dependent enzymes, pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase (alpha KGDH) and transketolase. Reductions in enzyme activities are present both in affected areas of AD brain as well as in relatively well conserved tissue. Decreased TDP concentrations and concomitantly increased TMP in autopsied brain tissue from AD patients and in CSF from patients with Dementia of the Alzheimer Type suggests that CNS thiamine phosphorylation-dephosphorylation mechanisms are disrupted in AD. alpha KGDH is a rate-limiting enzyme for cerebral glucose utilization and decreases in its activity are associated with lactic acidosis, cerebral energy failure and neuronal cell loss. Deficiencies of TDP-related metabolic processes could therefore participate in neuronal cell death mechanisms in AD.

Alzheimer Disease↗
PubMed1996

Regional distribution of binding sites for the nitric oxide synthase inhibitor L-[3H]nitroarginine in rat brain.

The regional distribution of NG-nitro-L-[(3)H]arginine (L-[(3)H]NOARG) binding to different regions of rat brain was studied by quantitative autoradiography. These studies revealed highest density of binding sites in cerebellum, anterior olfactory nucleus, islands of Calleja and substantia nigra with appreciable binding site densities in inferior colliculus, superior colliculus, olfactory tubercle and dorsal tegmental nucleus. The regional distribution of L-[(3)H]NOARG binding, is in good agreement with the distribution of nitric oxide synthase studied previously by NADPH-diaphorase staining and immunohistochemistry using antibodies against neuronal nitric oxide synthase. The kinetics of L-[(3)H]NOARG binding to the cytosolic preparations of cerebral cortex, cerebellum, hippocampus and striatum was studied using an in vitro binding technique. Specific L-[(3)H]NOARG binding was of nanomolar affinity, saturable, and best fit to a single-site model in all four brain regions. These studies support the potential use of L-[(3)H]NOARG binding as a tool for further elucidation of the regional distribution and functional properties of NOS in the central nervous system.

Animals↗
PubMed1996

Brain extracellular quinolinic acid in chronic experimental hepatic encephalopathy as assessed by in vivo microdialysis: acute effects of L-tryptophan.

Increased brain quinolinic acid (QUIN) levels have been suggested to play a role in hepatic encephalopathy (HE). Previous brain tissue studies have been unable to confirm this hypothesis. Because QUIn is a potent NMDA-receptor agonist, it also is relevant to determine brain extracellular QUIN levels in HE. For this purpose, we assessed frontal neocortical extracellular QUIN levels by in vivo microdialysis in rats subjected to a portacaval shunt (PCS). We also evaluated the acute effects of altered L-tryptophan (L-TRP) availability on brain extracellular QUIN levels. The basal extracellular L-TRP levels were significantly (p < .001) higher in the PCS rats than in the sham-operated controls. However, the QUIN level (p < .05) and the QUIN to L-TRP ratio (p < .01) were significantly lower in the PCS rats. Elevated L-TRP availability increased the QUIN levels to a similar degree in both sham and PCS rats. This study, in conjunction with our previous results, does thereby not support a major involvement of QUIN in the pathogenesis of HE.

Animals↗
PubMed1996

Ammonium acetate challenge in experimental chronic hepatic encephalopathy induces a transient increase of brain 5-HT release in vivo.

Ammonia has been shown to cause release of neurotransmitters such as serotonin (5-hydroxytryptamine; 5-HT) from synaptosomal preparations in vitro. In the present study, frontal neocortical extracellular levels of 5-HT and its major metabolite, 5-hydroxyindole-3-acetic acid (5-HIAA), were determined in vivo by the use of microdialysis in portacaval shunted (PCS) rats, an experimental model of chronic hepatic encephalopathy (HE), prior to and after an acute coma-inducing administration of ammonium acetate (NH4Ac; 5.2 mmol/kg, i.p.). PCS rats displayed elevated (P < 0.01) 5-HIAA but unaltered 5-HT extracellular levels compared with controls, supporting the contention of an increased neocortical 5-HT metabolism but unaltered neuronal 5-HT output in chronic HE. However, a transient elevation of extracellular 5-HT levels was observed when PCS-NH4Ac rats were in coma. Increased brain ammonia may thus augment neuronal 5-HT release in chronic HE, which in turn could be a causative for precipitation of more severe stages of HE.

Acetates↗
PubMed1996

Kinetics, pharmacology, and autoradiographic distribution of L-[3H]nitroarginine binding sites in rat cerebellum.

The kinetics and pharmacology of NG-nitro-L-[2,3,4,5-3H]arginine (L-[3H]NOARG) binding to rat cerebellum were investigated using in vitro radioligand binding. Specific L-[3H]NOARG binding in cerebellum was of nanomolar affinity, reversible, saturable, and best fit to a single-site model. Specific binding was Ca2+ dependent and sensitive to pH (with an optimum of 5.5-7.0). Added calmodulin (1.5-40 micrograms/ml) had no influence on specific L-[3H]NOARG binding. However, the calmodulin antagonists W-5, W-13, and calmidazolium inhibited L-[3H]-NOARG binding with IC50 values in the micromolar range, and calmodulin (10 micrograms/ml) competitively reversed this inhibition. Nitric oxide synthase (NOS) inhibitors (NG-nitro-L-arginine methyl ester and NG-monomethyl-L-arginine acetate) and L-arginine displaced L-[3H]NOARG binding with IC50 values in the nanomolar range, whereas D-arginine and basic amino acids (L-lysine and L-histidine) displaced L-[3H]NOARG binding with IC50 values in the millimolar range. A comparison of the NOS functional assay with L-[3H]NOARG binding in rat cerebellum showed similar profiles of Ca2+ dependency and inhibitory kinetics. Quantitative autoradiographic distribution of L-[3H]NOARG binding sites was significantly higher in the molecular layer than in the granular layer of cerebellum. These studies confirm the potential use of L-[3H]NOARG binding to study the regional distribution and functional properties of NOS.

Animals↗
PubMed1996

Alterations in serotonin parameters in brain of thiamine-deficient rats are evident prior to the appearance of neurological symptoms.

Biochemical alterations of serotoninergic parameters have been demonstrated in experimental thiamine deficiency. In addition, hypophagia and hypothermia, two physiological processes associated with changes in the serotonin [5-hydroxytryptamine (5-HT)] system, are manifest early during the progression of thiamine deficiency. The binding of selected 5-HT radioligands was therefore investigated in discrete brain regions of pyrithiamine-induced thiamine-deficient rats. Using quantitative receptor autoradiography, the binding of 8-hydroxy-2-(di-n-[3H]propylamino) tetralin, a ligand used to label the somatodendritic 5-HT1A autoreceptor of the dorsal raphe nucleus, was found to be unaffected in this region, suggesting that the structural integrity of the 5-HT cell bodies is maintained throughout the course of pyrithiamine treatment. Increased binding of [3H]-ketanserin was observed in regions considered vulnerable as well as in some considered to be nonvulnerable during the course of thiamine deficiency. These binding changes, which appear to represent changes in the density of the postsynaptic 5-HT2A receptor population rather than the "tetrabenazine-sensitive" vesicular monoamine transporter, are evident before the appearance of histopathologic lesions and coincide with altered tissue concentrations of 5-HT. These data suggest that 5-HT neurons, although structurally intact, are functionally affected early during the progression of thiamine deficiency. These alterations, which are likely a part of adaptive neuronal change consequent to thiamine dysfunction, may be important in the physiological manifestations and the learning deficits commonly encountered in experimental thiamine deficiency.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1996

L-[3H]nitroarginine and L-[3H]arginine uptake into rat cerebellar synaptosomes: kinetics and pharmacology.

Characteristics of the transport of the nitric oxide synthase substrate L-arginine and its inhibitor, NG-nitro-L-arginine (L-NOARG), into rat cerebellar synaptosomes were studied. Uptake of both L-arginine and L-NOARG was linear with increasing amount of protein (up to 40 micrograms) and time of incubation (up to 5 min) at 37 degrees C. Uptake of both compounds reached a steady state by 20 min. Maximal uptake of L-NOARG (650 pmol/mg of protein) was three to four times higher than that of L-arginine (170 pmol/mg of protein). L-NOARG uptake showed biphasic kinetics (Km1 = 0.72 mM, Vmax 1 = 0.98 nmol/min/mg of protein; Km2 = 2.57 mM, Vmax2 = 16.25 nmol/min/mg of protein). L-Arginine uptake was monophasic with a Km of 106 microM and a Vmax of 0.33 nmol/min/mg of protein. L-NOARG uptake was selectively inhibited by L-NOARG, NG-nitro-L-arginine methyl ester, and branched-chain and aromatic amino acids. L-Alanine and L-serine also inhibited L-NOARG uptake but with less potency. Uptake of L-arginine was selectively inhibited by NG-monomethyl-L-arginine acetate and basic amino acids. These studies suggest that in rat cerebellar synaptosomes, L-NOARG is transported by the neutral amino acid carrier systems T and L with high affinity, whereas L-arginine is transported by the basic amino acid carrier system y+ with high affinity. These data indicate that the concentration of competing amino acids is an important factor in determining the rates of uptake of L-NOARG and L-arginine into synaptosomes and, in this way, may control the activity of nitric oxide synthase.

Amino Acids↗
PubMed1996

Portacaval anastomosis results in altered neuron--astrocytic metabolic trafficking of amino acids: evidence from 13C-NMR studies.

13C-NMR spectroscopy was used to evaluate the dynamic consequences of portacaval anastomosis on neuronal and astrocytic metabolism and metabolic trafficking between neurons and astrocytes. Glutamate is predominantly labeled from [1-13C]glucose, whereas [2-13C]acetate is more efficient in labeling glutamine, in accordance with its primary metabolism in astrocytes. Alanine and succinate labeling was only observed with [1-13C]glucose as precursor. Brain [1-13C]glucose metabolism in portacaval-shunted rats was similar to that in sham-operated controls with the exception of labeled glutamine and succinate formation, which was increased in shunted rats. The 13C enrichment was, however, decreased owing to an increase in total glutamine and succinate. Using [2-13C]acetate, on the other hand, flux of astrocytic label to neurons was severely decreased because label incorporation into glutamate, aspartate, and GABA was decreased following portacaval shunting. The latter amino acids are predominantly localized in neurons. These findings demonstrate that metabolic trafficking of amino acids from astrocytes to neurons is impaired in portacaval-shunted rats.

Acetates↗
PubMed1996

Quantitative autoradiography using selective radioligands for central and peripheral-type benzodiazepine receptors in experimental Wernicke's encephalopathy: implications for positron emission tomography imaging.

Wernicke's encephalopathy (WE) is difficult to diagnose during life, with up to 80% of cases being missed by routine neurological evaluation in both alcoholics and AIDS patients. Therefore, there is a need for noninvasive diagnostic procedures. Using the pyrithiamine-treated rat, an animal model of WE, we have studied, using radioligands for central (neuronal) and "peripheral-type" (glial) benzodiazepine receptors, the regional distribution of changes in the densities of these receptors in relation to the degree of reactive gliosis accompanying neuronal loss. Histological studies revealed neuronal loss in selective regions, including the thalamus, inferior colliculus, inferior olivary nucleus, and mammillary body. Autoradiographic studies demonstrated increases in densities of [3H]PK11195 binding sites that closely paralleled the topographic distribution of neuronal cell loss and reactive gliosis. In contrast, [3H]Ro15-178 showed poor spatial correlation, with the neuronal loss seen in pyrithiamine-induced thiamine deficiency. The positron emission tomography ligand [11C]PK11195 may be useful for the assessment of thiamine deficiency-induced brain damage in human alcoholics.

Animals↗
PubMed1996

Central muscarinic cholinergic M1 and M2 receptor changes in congenital ornithine transcarbamylase deficiency.

Congenital ornithine transcarbamylase (OTC) deficiency results in neuropathologic damage to the cerebral cortex, basal ganglia, and thalamus. However, the precise nature of the cell loss, as well as the pathophysiologic mechanisms responsible for it, have not been fully elucidated. In the present study, densities of the M1 and M2 subclasses of muscarinic cholinergic binding sites were assessed using quantitative receptor autoradiography in the brains of sparse-fur (spf) mice with congenital OTC deficiency and in age-matched CD-1 controls. Densities of binding sites for the muscarinic M1 subtype ligand [3H]pirenzepine were reduced by 24-54% (p < 0.01) in frontal cortex, caudate/ putamen, and hippocampal CA1 and CA2 areas. Since muscarinic M1 sites are localized presynaptically, their selective loss, together with a previous report of reduced activities of the presynaptic cholinergic enzyme choline acetyltransferase, confirms that loss of cholinergic neurons is an important feature of congenital OTC deficiency. Densities of binding sites for the predominantly postsynaptic muscarinic M2 subtype ligand 3H-AFDX 384 were increased by up to 60% (p < 0.01) in cerebral cortex, hippocampus, globus pallidus, as well as thalamic and hypothalamic structures of OTC-deficient mice. Increased M2 sites in the cerebral cortex, hippocampus, and thalamus are most likely the result of up-regulation of these sites after the loss of the presynaptic neuron. These findings support the presence of a central muscarinic cholinergic lesion in congenital OTC deficiency.

Animals↗
PubMed1995

Na+,K(+)-ATPase activities are increased in brain in both congenital and acquired hyperammonemic syndromes.

Activities of Na+,K(+)-ATPase were measured in brain regions of experimental animals with either congenital or acquired hyperammonemia. In the sparse-fur (spf) mutant mouse, with a genetic X-linked deficiency of ornithine transcarbamylase, an animal model of congenital hyperammonemia, Na+,K(+)-ATPase was increased in frontal cortex (by 57%, P < 0.001), cerebellum (by 61%, P < 0.001), brainstem (by 71%, P < 0.001) and striatum (by 48%, P < 0.01). Four weeks following portacaval anastomosis in the rat, Na+,K(+)-ATPase activities were increased in cerebellum and striatum (by 19%, P < 0.01) and in brainstem (by 28%, P < 0.01). Stimulation of Na+,K(+)-ATPase and the subsequent alteration of neuronal excitability could contribute to the CNS dysfunction characteristic of chronic hyperammonemic syndromes.

Ammonia↗
PubMed1995

A quantitative autoradiographic study of muscarinic cholinergic receptor subtypes in the brains of pyrithiamine-treated rats.

Previous studies describe decreased acetylcholine synthesis in brain as well as neurobehavioral evidence for a central muscarinic cholinergic deficit in pyrithiamine-induced thiamine-deficient rats. In order to further evaluate this possibility, quantitative autoradiographic procedures using [3H]quinuclidinyl benzilate (for total muscarinic binding sites), [3H]pirenzepine (for muscarinic M1 sites) and [3H]AF-DX 384 (for muscarinic M2 sites) were performed at early (presymptomatic) and late (symptomatic) stages of thiamine deficiency induced in rats by administration of the central thiamine antagonist, pyrithiamine. No significant alterations in densities of M1, M2 or total muscarinic binding sites were observed in any brain structure evaluated at either early or late stages of thiamine deficiency. These findings do not support a major role for modifications of muscarinic cholinergic function in the pathogenesis of the neurological symptoms of thiamine deficiency.

Animals↗
PubMed1995

Effect of probenecid on 5-hydroxyindoleacetic acid in cisternal cerebrospinal fluid of rats with portacaval anastomosis.

Portal-systemic encephalopathy (PSE) is characterized by a neuropsychiatric disorder progressing through personality changes, to stupor and coma. Previous studies have revealed alterations of serotonin and of its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in brain tissue and CSF in experimental (rat) and human PSE. Increased brain 5-HIAA concentrations could result from its decreased removal rather than to increased serotonin metabolism. In order to evaluate this possibility, CSF 5-HIAA concentrations were measured using an indwelling cisterna magna catheter technique at various times following end-to-side portacaval anastomosis in rats (the most widely used animal model of PSE) treated with probenecid, a competitive inhibitor that blocks the active transport of acid metabolites out of the brain and CSF. Following portacaval anastomosis and probenecid treatment, CSF concentrations of 5-HIAA were increased to a greater extent than in sham-operated controls. When data were expressed as per-cent baseline values, the relative increase of CSF 5-HIAA in portacaval shunted rats following probenecid treatment was not significantly different from sham-operated controls. These findings confirm that increased 5-HIAA in the CNS in experimental PSE results from increased 5HT metabolism or turnover and that the probenecidsensitive acid metabolite carrier is intact in PSE.

Animals↗
PubMed1995

Regional alterations of dopamine and its metabolites in rat brain following portacaval anastomosis.

Hyperammonemia and changes in brain monoamine metabolism have been proposed to contribute to the pathogenesis of the neuropsychiatric symptoms characteristic of human portal-systemic encephalopathy (PSE) resulting from chronic liver disease. Portacaval anastomosis (PCA) in the rat leads to sustained hyperammonemia and mild encephalopathy. In order to evaluate the role of dopamine (DA) metabolism in PSE, levels of DA and its metabolites were measured by HPLC with electrochemical detection in brain regions of rats with PCA at various stages of encephalopathy precipitated by ammonium acetate administration. Following ammonium acetate administration, rats with PCA rapidly develop severe neurological signs of encephalopathy progressing through loss of righting reflex to coma; sham-operated control animals administered ammonium acetate showed no such neurological deterioration. Concentrations of the DA metabolites DOPAC and HVA as well as [DA metabolites]/[DA] ratios, an indirect measure of DA turnover in brain, were increased in caudate-putamen, in cingulate and pyriform entorhinal cortices as well as in raphe nucleus and locus coeruleus. Increased DA metabolites, however, did not worsen at coma stages of PSE. Increased DA turnover thus appears to relate to early neuropsychiatric and extrapyramidal symptoms of PSE.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

Regional alterations of thiamine phosphate esters and of thiamine diphosphate-dependent enzymes in relation to function in experimental Wernicke's encephalopathy.

Thiamine phosphate esters (thiamine monophosphate-TMP; thiamine diphosphate-TDP and thiamine triphosphate-TTP) were measured as their thiochrome derivatives by High Performance Liquid Chromatography in the brains of pyrithiamine-treated rats at various stages during the development of thiamine deficiency encephalopathy. Severe encephalopathy was accompanied by significant reductions of all three thiamine phosphate esters in brain. Neurological symptoms of thiamine deficiency appeared when brain levels of TMP and TDP fell below 15% of normal values. Activities of the TDP-dependent enzyme alpha-ketoglutarate dehydrogenase were more severely reduced in thalamus compared to cerebral cortex, a less vulnerable brain structure. On the other hand, reductions of TTP, the non-cofactor form of thiamine, occurred to a greater extent in cerebral cortex than thalamus. Early reductions of TDP-dependent enzymes and the ensuing metabolic pertubations such as lactic acidosis impaired brain energy metabolism, and NMDA-receptor mediated excitotoxicity offer rational explanations for the selective vulnerability of brain structures such as thalamus to the deleterious effects of thiamine deficiency.

Animals↗