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Brian Dean

Publications and source records attributed to Brian Dean.

36 records · Page 2Linked to original sources
PubMed2004

Glucuronidation, oxidative metabolism, and bioactivation of enterolactone in rhesus monkeys.

Enterolactone (ENL) and enterodiol (END) are mammalian lignans derived from the plant lignans matairesionol (MAT), secoisolariciresinol (SECO), and other dietary precursors. ENL was found to undergo extensive glucuronidation with rhesus liver microsomes to form O-glucuronides at both phenolic hydroxy groups. In addition to glucuronidation, ENL was found to be a good substrate for oxidative metabolism. The major products had a m/z of 313 or 295 by LC-MS analysis in negative ion mode and were determined to be products of monohydroxylation of ENL. The m/z 295 products were the result of a dehydration of the m/z 313 in the MS source. Addition of N-acetylcysteine (NAC) to the NADPH incubations resulted in a decrease of at least 2 major monohydroxylated products and the formation of a major and several minor new products with a m/z of 474. The major adduct was isolated, purified for NMR, and confirmed to be the NAC adduct of the ENL catechol. Incubations of ENL with liver microsomes containing UDPGA, NADPH, and NAC resulted in the formation of ENL-glucuronides; no NAC adducts were detected by LC-MS. Incubations of ENL with human and rhesus hepatocytes resulted in several metabolites. The major metabolites in hepatocytes were the glucuronic acid conjugates; minor amounts of the sulfate conjugate(s) and monohydroxylated products were also detected by LC-MS. Glutathione or other thiol adducts were not detected in hepatocytes. Conclusion. The high efficiency and specificity for the glucuronidation of ENL decrease its potential toxicity via CYP450 bioactivation.

4-Butyrolactone↗
PubMed2004

Expression of truncated presenilin 2 splice variant in Alzheimer's disease, bipolar disorder, and schizophrenia brain cortex.

We have analysed the expression of a truncated variant presenilin 2 protein (PS2V) in frontal cortex from subjects with Alzheimer's disease (AD) and age-matched controls, and compared these results with cortex from bipolar disorder (BP), schizophrenia (SZ) and controls in a second brain bank collection. PS2V protein was detected as a 14 kDa species with antibodies directed to the PS2 N-terminal region and to the new C-terminus created by alternative transcription. PS2V protein levels were significantly increased by two-fold in AD cortex, as compared to age-matched controls. In tissue from the second collection, levels of PS2V were markedly elevated in some BP and SZ cases, but there was no overall difference between diagnostic groups. Our findings support previous evidence for increased expression of this variant PS2 isoform in sporadic AD and suggest this isoform may contribute to neurodegeneration.

Adult↗
PubMed2004

Identification of UGT2B9*2 and UGT2B33 isolated from female rhesus monkey liver.

Two UDP-glucuronosyltransferases (UGT2B9(*)2 and UGT2B33) have been isolated from female rhesus monkey liver. Microsomal preparations of the cell lines expressing the UGTs catalyzed the glucuronidation of the general substrate 7-hydroxy-4-(trifluoromethyl)coumarin in addition to selected estrogens (beta-estradiol and estriol) and opioids (morphine, naloxone, and naltrexone). UGT2B9(*)2 displayed highest efficiency for beta-estradiol-17-glucuronide production and did not catalyze the glucuronidation of naltrexone. UGT2B33 displayed highest efficiency for estriol and did not catalyze the glucuronidation of beta-estradiol. UGT2B9(*)2 was found also to catalyze the glucuronidation of 4-hydroxyestrone, 16-epiestriol, and hyodeoxycholic acid, while UGT2B33 was capable of conjugating 4-hydroxyestrone, androsterone, diclofenac, and hyodeoxycholic acid. Three glucocorticoids (cortisone, cortisol, and corticosterone) were not substrates for glucuronidation by liver or kidney microsomes or any expressed UGTs. Our current data suggest the use of beta-estradiol-3-glucuronidation, beta-estradiol-17-glucuronidation, and estriol-17-glucuronidation to assay UGT1A01, UGT2B9(*)2, and UGT2B33 activity in rhesus liver microsomes, respectively.

Amino Acid Sequence↗
PubMed2004

The neurobiology of bipolar disorder: findings using human postmortem central nervous system tissue.

OBJECTIVE: Postmortem brain studies have been undertaken to understand changes in the molecular architecture of the central nervous system (CNS) of subjects with bipolar disorder. These studies, along with a limited number of functional neuroimaging studies, have been reviewed to provide information on the neurobiology underlying the disorder. METHOD: Findings from the study of postmortem brain tissue and neuroimaging were reviewed if their focus was on the molecular architecture of the human CNS to identify future lines of research required to understand the underlying pathology of bipolar disorder. RESULTS: There is considerable evidence to implicate the serotonergic system of the CNS and abnormalities in signal transduction pathways in the pathology of bipolar disorder. In addition, preliminary findings suggest that changes in the benzodiazepine binding site on the gamma aminobutyric acidA receptor may be affected in bipolar disorder. CONCLUSIONS: Further systematic studies on the serotonergic systems of the CNS, as well as the interaction between neurotransmitter receptors, G-proteins and signal transduction pathways are required to better understand the pathology of bipolar disorder. In addition, findings on the serotonin transporter indicate that changes in presynaptic function may be a critical component of the pathology of bipolar disorder.

Binding Sites↗
PubMed2003

Increased levels of apolipoprotein E in the frontal cortex of subjects with schizophrenia.

BACKGROUND: It is unclear whether altered expression of a specific isoform of apolipoprotein E (apoE) is associated with the pathology of schizophrenia. METHODS: To address whether apoE may be involved in the pathology of schizophrenia, we measured the genotypic and allelic frequency of polymorphisms in its gene and transcriptional regulatory region in DNA from Brodmann's area (BA) 9 obtained postmortem from schizophrenic and control subjects as well as its levels in the same tissue using Western blot analysis. RESULTS: The genotypic or allelic frequencies of any polymorphism studied did not vary between diagnostic cohorts. There was a significant increase in the levels of apoE protein in BA 9 from the schizophrenic subjects (Mean +/- SEM: 270 +/- 8.3 vs. 238 +/- 7.1 ng apoE/mg protein, p =.008) and a decrease in tissue from an analogous cortical region from rats treated with haloperidol compared with vehicle-treated animals (50 +/- 6.4 vs. 116 +/- 9.2 ng apoE/mg protein; p =.0002). CONCLUSIONS: These data support the hypothesis that increased levels of apoE may be associated with the pathology of schizophrenia and that antipsychotic drugs decrease apoE levels as part of their therapeutic actions.

Adult↗
PubMed2003

Apolipoprotein D levels are elevated in prefrontal cortex of subjects with Alzheimer's disease: no relation to apolipoprotein E expression or genotype.

BACKGROUND: Apolipoprotein E (apoE) has been implicated in the pathology of AD ever since inheritance of the epsilon4 allele was shown to be an important risk factor for the development of AD. Apolipoprotein D (apoD) is elevated in association with several central nervous system disorders, including Alzheimer's disease (AD), and has been proposed to be an especially robust marker for brain regions specifically affected by particular neuropathologies. Progressive cognitive decline is the core clinical feature of AD and is associated with disturbances in the prefrontal cortex. METHODS: We measured apoD levels in prefrontal cortex samples obtained postmortem from 20 autopsy-confirmed AD subjects and 40 control subjects. RESULTS: Enzyme-linked immunosorbent assay analysis revealed a significant increase in apoD expression in AD subjects compared with control subjects (.218+/-.029 microg/mg protein vs.117+/-.011 microg/mg protein; p=0003). There was no significant difference in apoD expression between early-onset and late-onset Alzheimer's subjects. Apolipoprotein D expression levels were not correlated with apoE levels, nor were they correlated with inheritance of the APOE epsilon4 allele. CONCLUSIONS: These findings suggest that apoD may be related to the cognitive decline observed in AD patients and that apoD and apoE likely play different roles in the pathogenesis of AD.

Aged↗
PubMed2003

[3H]RX 821002 in human dorsolateral prefrontal cortex: no changes in postmortem tissue from subjects with schizophrenia.

One of the major differences between the atypical antipsychotic drugs clozapine and olanzapine is that clozapine has a two-fold higher affinity for the alpha(2)-adrenoreceptors. As clozapine can have therapeutic benefits in individuals that do not respond to other antipsychotic drugs, this raises the possibility that changes in the alpha(2)-adrenoreceptors could be a marker for a predisposition to treatment resistance. A methodology has been optimised to measure the binding of [3H]RX 821002 to alpha(2)-adrenoreceptors in human postmortem CNS and has shown that these receptors are not altered in Brodmann's area 9 from subjects with schizophrenia. These data add to those of one other study that showed the alpha(2)-adrenoreceptors were not altered in Brodmann's area 10 and the hippocampus from subjects with schizophrenia, and do not support the hypothesis that changes in alpha(2)-adrenoreceptors are a marker for treatment resistance in schizophrenia.

Adrenergic alpha-2 Receptor Antagonists↗
PubMed2003

Cannabis-sensitive dopaminergic markers in postmortem central nervous system: changes in schizophrenia.

BACKGROUND: This study investigated if changes in pre-synaptic markers on dopaminergic neurons (dopamine transporter [DAT], tyrosine hydroxylase [TH]) were present in the caudate from subjects with schizophrenia who had Delta(9)(-)tetrahydrocannabinol (THC) in their blood at autopsy. These changes were posited because animal studies show that treatment with THC decreases dopamine uptake and TH in the striatum. METHODS: Studies utilized caudate, obtained postmortem, from 14 schizophrenic and 14 control subjects. [(3)H]mazindol binding to caudate, measured using autoradiography, was taken as a measure of DAT; TH levels were estimated using an antihuman TH antibody and Western blotting. RESULTS: There was decreased [(3)H]mazindol binding to DAT in the caudate from the schizophrenic subjects with no detectable blood THC levels (THC(-)) compared with THC(-) control subjects (mean +/- SEM: 240 +/- 19 vs. 296 +/- 14 fmol/mg estimated tissue equivalents, p =.01). There were no significant differences between levels of DAT in the caudate from schizophrenic and control subjects that had THC in their blood. Tyrosine hydroxylase was not different in any diagnostic cohort. CONCLUSIONS: Our data suggests that DAT is decreased in the caudate from THC(-) subjects with schizophrenia, a change that may be reversed by ingesting THC from cannabis.

Adult↗
PubMed2003

Studies on serotonergic markers in the human hippocampus: changes in subjects with bipolar disorder.

BACKGROUND: Various studies suggest the hippocampus and serotonergic systems are important in the pathology of bipolar disorder (BD). We therefore measured hippocampal serotonergic markers in post-mortem tissue from BD and control subjects. METHODS: The density and affinity of [3H]citalopram binding to the serotonin transporter (SERT), as well as the density of the 5HT(2A), 5HT(1A), 5HT(1D) and 5HT(1F) receptors were measured. RESULTS: The density of SERT and 5HT receptors was no different in BD. There was a significant decrease in the affinity of [3H]citalopram binding to SERT in the stratum lacunosum-moleculare (S(lac)) in BD (K(d) mean+/-S.E.M.=4.3+/-0.8 vs. 1.9+/-0.3 nM). LIMITATIONS: This study was completed using relatively small cohorts. CONCLUSIONS: There are no generalised changes in hippocampal serotonergic markers in the hippocampus from subjects with BD. There is a decreased affinity of radioligand binding to S(lac) SERT in subjects with BD.

Affinity Labels↗
PubMed2003

The cortical serotonin2A receptor and the pathology of schizophrenia: a likely accomplice.

A large body of evidence shows that there is a change in the density of cortical serotonin2A receptors (5HT2AR) in post-mortem CNS from subjects with schizophrenia. Furthermore, some antipsychotic drugs have also been shown to cause a decrease in the density of 5HT2AR in the rat CNS. Thus, it appeared possible that changes in this receptor in human post-mortem CNS simply reflected an antipsychotic drug effect. However, a great deal of research on the 5HT2AR and schizophrenia now suggests that the changes in this receptor are complex and may be involved in both the pathology of the disorder and the effects of some antipsychotic drugs. Moreover, recent advances in basic research on the role of the 5HT2AR in the CNS add further support to the hypothesis that the receptor could be involved in the pathology of the illness. In particular, an argument will be developed that the changes in the 5HT2AR in schizophrenia are reflective of a real or perceived change in serotonergic tone and that this forms an important part of the pathology of the illness.

Animals↗
PubMed2002

The heterogeneity of central benzodiazepine receptor subtypes in the human hippocampal formation, frontal cortex and cerebellum using [3H]flumazenil and zolpidem.

The ability of clonazepam and zolpidem to displace [3H]flumazenil binding was measured in the human hippocampal formation, frontal cortex (BA9) and the cerebellum using in situ radioligand binding and autoradiography. The use of high resolution phosphorimaging in all regions indicated the displacement of [3H]flumazenil by clonazepam was monophasic with K(i) values ranging from 2.73+/-0.17 to 6.49+/-0.21 nM. [3H]flumazenil binding that was not displaced by clonazepam ranged from 3.39+/-0.86 to 7.15+/-1.11%. The ability of zolpidem to displace [3H]flumazenil was also monophasic in the frontal cortex and cerebellum with K(i) values of 37.53+/-1.79 and 31.80+/-1.68 nM, respectively. In contrast, within all hippocampal regions, zolpidem displacement of [3H]flumazenil was biphasic, with K(i) values for the high affinity site ranging from 0.13+/-0.04 to 0.54+/-0.03 nM, whereas the low affinity site was between 84.98+/-1.58 and 98.84+/-1.89 nM. In addition, zolpidem insensitive [3H]flumazenil binding was observed to vary markedly between brain regions, ranging between 37.85+/-1.60 and 6.13+/-0.83%. In conclusion, the present results indicate that in situ radioligand binding and high-resolution phosphorimaging techniques can be utilized to measure the differential displacement of [3H]flumazenil by zolpidem and clonazepam. Moreover, our data suggests that the differential distribution of the zolpidem insensitive component of [3H]flumazenil binding is an indicator of GABA/BZ receptors assembled by different subunits within the human brain.

Cerebellum↗
PubMed2002

Isolation and characterization of a UDP-glucuronosyltransferase (UGT1A01) cloned from female rhesus monkey.

An isoform (rhesus UGT1A01) orthologus to the human UGT1A1 was cloned and sequenced from female rhesus monkey liver cDNA using primers designed from the human nucleotide sequences. Open reading frame analysis of the PCR-generated product encodes a 533-amino acid protein with a proposed 27-residue signal peptide. Nucleotide sequence comparison of rhesus UGT1A01 to other rhesus UGT1A isoforms detected a single-transition mutation at nucleotide 1520 (T-->C), resulting in a neutral F to S substitution at position 507. Rhesus UGT1A01 was greater than 99 and 95% identical to cynomolgus UGT1A01 and human UGT1A1, respectively. The rhesus UGT1A01 was expressed in HK-293 cells for functional analysis. Catalytic activity of UGT1A01 was determined with 7-hydroxy-4-(trifluoromethyl)-coumarin and more specific human UGT1A1 substrates (1-naphthol, beta-estradiol, 17 alpha-ethinylestradiol, and bilirubin). Expression of UGT1A01 protein was also detected by a Western blot utilizing a polyclonal antibody developed against the human UGT1A family.

Amino Acid Sequence↗
PubMed2002

High-resolution phosphor imaging: validation for use with human brain tissue sections to determine the affinity and density of radioligand binding.

This study investigated the suitability of high-resolution storage phosphor imaging for the quantitative analysis of radioligand binding to human brain tissue. Hence, the binding of [(3)H]mazindol to the dopamine transporter in caudate-putamen tissue homogenates or frozen tissue sections apposed to either autoradiographic film or phosphor imaging plates was measured. Estimates of binding affinity were similar for homogenate studies and phosphor imaging plates (Kd=6.44+/-0.14 and 6.91+/-0.47 nM, respectively), but higher values were obtained with film autoradiography (Kd=11.31+/-0.82 nM). The density of binding was similar for both autoradiographic techniques (Bmax=371.9+/-30.8 fmol/mg estimated tissue equivalent, ETE (imaging plate) and 425+/-13.77 fmol/mg ETE (film)), although lower values were obtained from tissue homogenates (Bmax=64.27+/-6.74 fmol/mg wet weight). These results suggest that high resolution phosphor imaging can be used to analyse radioligand binding parameters in human brain tissue. Moreover, the reduced exposure time of phosphor imaging plates (e.g. 7 days vs 5 weeks) allows results to be obtained more rapidly than with conventional film autoradiography.

Adult↗
PubMed2002

Decreased density of [3H]TCP binding following antipsychotic drug withdrawal in rats.

Antipsychotic drugs have been reported to increase the expression of subunits of the NMDA receptor at the level of mRNA but it is not clear whether such effects are apparent at the level of the radioligand binding or receptor protein. Therefore, we examined the effect of treatment of, and withdrawal from, haloperidol, chlorpromazine, olanzapine or clozapine on the binding of [3H]N-[1-(2-thienyl)cyclohexyl]piperidine ([3H]TCP ) to the open ion channel of the NMDA receptor in rat caudate-putamen, hippocampus and frontal cortex. [3H]TCP binding was not significantly different in the caudate-putamen, hippocampus and cortex after three months of treatment with any antipsychotic drug. There were significant decreases in [3H]TCP binding in rat caudate-putamen and cortex, but not hippocampus, one month after ceasing treatment. Decreases in the caudate-putamen were detected in rats previously treated with chlorpromazine (0.1 mg/kg/day) and clozapine (0.1 and 1.0 mg/kg/day). In the cortex, decreases in [3H]TCP binding were also detected in rats previously treated with olanzapine (0.1 mg/kg/day) for three months. These data suggest that changes in the NMDA receptor associated ion channels occur following antipsychotic drug withdrawal.

Animals↗
PubMed2002

[(3)H]Flumazenil binding in the human hippocampal formation, frontal cortex and cerebellum detected by high-resolution phosphorimaging.

The pharmacological characterisation of the benzodiazepine binding site associated with the gamma-aminobutyric acid (GABA(A)) receptor in human brain has been demonstrated using in situ radioligand binding and autoradiography. The use of high-resolution phosphorimaging has allowed both the affinity (K(d)) and density (B(max)) of [(3)H]flumazenil binding to be measured within regions of the hippocampal formation as well as the cerebellum and frontal cortex. The Scatchard plots of data from all brain regions were linear with Hill coefficients close to unity consistent with the presence of a single binding site for [(3)H]flumazenil. The affinities of [(3)H]flumazenil binding within all the brain regions were similar (K(d) 1.57+/-0.20-3.08+/-0.01 nM), while the density of [(3)H]flumazenil binding varied significantly between the brain regions analysed (B(max) 182.7+/-7.3-596.7+/-34.0 fmol/mg ETE; P<0.0001). In conclusion, the present results indicate that in situ radioligand binding and high-resolution phosphorimaging techniques can be utilized to measure the distribution, density and affinity of [(3)H]flumazenil to the GABA(A) receptor within the human frontal cortex, cerebellum and hippocampal formation.

Animals↗
PubMed2002

Changes in the molecular structure of the brain in bipolar disorder: findings using human postmortem brain tissue.

Human postmortem brain studies are beginning to suggest mechanisms that underlie the pathology of bipolar disorder. Evidence that implicates abnormalities in G-protein, neurotransmitter turnover, neurotransmitter receptors and neurotransmitter receptor signal amplification will be reviewed. In addition, individual studies that suggest roles for pre-synaptic proteins, neuronal guidance proteins, neuropeptides, intra-neuronal ion homeostasis and environmental factor reactive proteins in the pathology of the disorder are also presented. Together these studies confirm that the pathology of bipolar disorder is complex but is yielding to intense study of the molecular architecture of postmortem brain tissue from subjects with the illness.

Bipolar Disorder↗