Benzodiazepine Receptor Binding Research Articles

Neonatal Clonazepam Administration Induced Long-Lasting Changes in GABAA and GABAB Receptors.

Benzodiazepines (BZDs) are widely used in patients of all ages. Unlike adults, neonatal animals treated with BZDs exhibit a variety of behavioral deficits later in life; however, the mechanisms underlying these deficits are poorly understood. This study aims to examine whether administration of clonazepam (CZP; 1 mg/kg/day) in 7–11-day-old rats affects Gama aminobutyric acid (GABA)ergic receptors in both the short and long terms. Using RT-PCR and quantitative autoradiography, we examined the expression of the selected GABAA receptor subunits (α1, α2, α4, γ2, and δ) and the GABAB B2 subunit, and GABAA, benzodiazepine, and GABAB receptor binding 48 h, 1 week, and 2 months after treatment discontinuation. Within one week after CZP cessation, the expression of the α2 subunit was upregulated, whereas that of the δ subunit was downregulated in both the hippocampus and cortex. In the hippocampus, the α4 subunit was downregulated after the 2-month interval. Changes in receptor binding were highly dependent on the receptor type, the interval after treatment cessation, and the brain structure. GABAA receptor binding was increased in almost all of the brain structures after the 48-h interval. BZD-binding was decreased in many brain structures involved in the neuronal networks associated with emotional behavior, anxiety, and cognitive functions after the 2-month interval. Binding of the GABAB receptors changed depending on the interval and brain structure. Overall, the described changes may affect both synaptic development and functioning and may potentially cause behavioral impairment.

Protective Effects of Artemisia persica Essential Oil Against Pentylenetetrazol-Induced Seizure in Male Mice with Emphasizing its Mechanism of Action

Background: A new approach in the treatment of epilepsy is to use new drugs with neuroprotective, antioxidant, and anti-inflammatory effects. Objectives: The study aimed to investigate the protective effects of Artemisia persica essential oil (EO) against pentylenetetrazol (PTZ)-induced seizure in mice. Methods: This experimental study was conducted at the Izeh Islamic Azad University, Iran. 70 male BALB/c mice were divided into seven groups of 10 using simple random allocation, including control (normal saline), PTZ (35 mg/kg i.p. with 48 hours intervals and then 60 mg/kg on the 10th day), interventions (PTZ plus daily i.p. injection of EO at doses of 50, 75, and 100 mg/kg), diazepam (PTZ plus EO at a dose of 100 mg/kg + diazepam), and flumazenil (PTZ plus EO at a dose of 100 mg/kg + flumazenil) groups. Results: The treatment of PTZ-kindled mice with 50 mg/kg of EO significantly reduced the seizure onset latency (P < 0.05). EO at a dose of 100 mg/kg significantly decreased tonic seizures, head tics, and repeated spinning and jumping (P < 0.05). Diazepam improved, and flumazenil weakened the anticonvulsant effects of the EO. The treatment of PTZ-kindled mice with EO (100 mg/kg) significantly decreased nitric oxide and malondialdehyde, and increased the total antioxidant capacity in both serum and brain (P < 0.05). EO at a dose of 100 mg/kg could significantly decrease IL-1β and TNF-α expression in the brain of epileptic mice (P < 0.05). Conclusions: A. persica EO shows anticonvulsant effects through benzodiazepine receptor binding activity and modulation of oxidative stress and the inflammatory process.

123I-Iomazenil Single-Photon Emission Computed Tomography Imaging in a Patient with Mild Traumatic Subdural Hematoma Accompanied by Delayed Transient Aphasia

Early and late images of 123I-iomazenil(IMZ)single-photon emission computed tomography(SPECT)reflect distributions of cerebral blood flow and those of cortical benzodiazepine receptor binding potential, respectively. Crossed cerebellar diaschisis reflects left-to-right asymmetry of metabolism in the cerebral hemispheres. We present a case of a 67-year-old woman who developed transient aphasia 3 days after the onset of a mild acute subdural hematoma. Computed tomography scan and magnetic resonance imaging during aphasia did not show enlargement of the hematoma or any new lesions. Electroencephalography did not show any abnormalities. Early images of 123I-IMZ SPECT 3 days after the onset of aphasia revealed a decrease in radioactivity in the right cerebellar hemisphere relative to that in the left cerebellar hemisphere. Late images of the same 123I-IMZ SPECT displayed a decrease in radioactivity in the left cerebral hemisphere relative to that in the right cerebral hemisphere. Twenty-four days later, the aphasia disappeared and the left-to-right asymmetries of radioactivity in the cerebellar and cerebral hemispheres on the early and late 123I-IMZ SPECT images also resolved.

Altered cerebral benzodiazepine receptor binding in post-traumatic stress disorder

Agonists of the γ-aminobutyric acid (GABA) type A benzodiazepine (BZD) receptor exert anxiolytic effects in anxiety disorders, raising the possibility that altered GABA-ergic function may play a role in the pathophysiology of anxiety disorders, such as post-traumatic stress disorder (PTSD). However, few neuroimaging studies have assessed the function or binding potential of the central GABAA BZD receptor system in PTSD. Therefore, our aim was to compare the BZD receptor binding potential between PTSD patients and healthy controls. Twelve medication-free participants with a current diagnosis of PTSD and 15 matched healthy controls underwent positron emission tomography (PET) imaging using [11C] flumazenil. Structural magnetic resonance imaging (MRI) scans were obtained and co-registered to the PET images to permit co-location of neuroanatomical structures in the lower resolution PET image data. Compared to healthy controls, PTSD patients exhibited increased BZD binding in the caudal anterior cingulate cortex and precuneus (p’s < 0.05). Severity of PTSD symptoms positively correlated with BZD binding in the left mid- and anterior insular cortices. This study extends previous findings by suggesting that central BZD receptor system involvement in PTSD includes portions of the default mode and salience networks, along with insular regions that support interoception and autonomic arousal.

Maturational Changes of Gamma-Aminobutyric Acid A Receptors Measured With Benzodiazepine Binding of Iodine 123 Iomazenil Single-Photon Emission Computed Tomography

BackgroundIodine 123 (I-123) iomazenil is a specific ligand of the central benzodiazepine receptor, which is a part of the postsynaptic gamma-aminobutyric acid A receptor complex. We performed statistical image processing of I-123 iomazenil single-photon emission computed tomography to elucidate maturational changes in the GABAergic system. MethodsThirty patients (18 boys and 12 girls, aged 17 days to 14 years) with cryptogenic focal epilepsy were enrolled and underwent I-123 iomazenil single-photon emission computed tomography. We used a semiquantitative analytical method consisting of brain surface extraction, anatomic normalization, and a three-parameter exponential model. We then assessed developmental changes in benzodiazepine receptor binding activity in 18 regions of interest in both hemispheres. ResultsThe highest benzodiazepine receptor binding activity was observed during early infancy in all regions of interest. Benzodiazepine receptor binding activity then decreased exponentially across development. Benzodiazepine receptor binding in the primary sensorimotor cortex, primary visual cortex, cerebellar vermis, and striatum declined more rapidly than that in the cerebellar hemispheres and the frontal cortex. The pons and the thalamus had the lowest benzodiazepine receptor binding activities during the neonatal period, and benzodiazepine receptor binding in these areas declined gradually after infancy toward adolescence. There were no differences in adjusted benzodiazepine receptor binding activity according to laterality or sex. ConclusionsBenzodiazepine receptor binding activity decreased exponentially during infancy in all regions of interest. Binding activity in the primary somatosensory and motor cortices (M1 and S1), the primary and association visual areas, the cerebellar vermis, and the striatum (caudate nucleus and putamen) tended to decline more rapidly than that in the cerebellar hemisphere and the frontal association cortex.

Increased Binding Potential of Brain Adenosine A1 Receptor in Chronic Stages of Patients with Diffuse Axonal Injury Measured with [1-methyl-11C] 8-dicyclopropylmethyl-1-methyl-3-propylxanthine Positron Emission Tomography Imaging.

The positron emission tomography (PET) radioligand for adenosine A1 receptor (A1R) [1-methyl-11C] 8-dicyclopropylmethyl-1-methyl-3-propylxanthine (MPDX) has recently been developed for human brain imaging. In the present study, we evaluated the alteration of the A1R in patients with diffuse axonal injury (DAI) in chronic stage in vivo. Ten patients with DAI (7 men and 3 women) were included in this study. Three PET examinations were sequentially performed to measure A1R binding with 11C-MPDX, glucose metabolism with 18F-fluorodeoxyglucose (FDG), and central benzodiazepine receptor binding with 11C-flumazenil (FMZ), and decreases of 11C-FMZ uptake indicate neuronal loss. 11C- MPDX did not depict any lesion with significantly decreased nondisplaceable binding potential (BPND) in comparison to healthy controls (14 men) in region of interest (ROI) analysis. Instead, it showed a significant increase of BPND in the lower frontal and posterior cingulate cortexes and rolandic area (p < 0.05) in ROI analysis. In 18F-FDG PET, the standardized uptake values (SUVs) ratio to the whole brain were decreased in anterior and posterior cingulate gyrus compared to controls (14 men and 9 women; p < 0.01). In 11C-FMZ PET, the SUV ratio to the cerebellum was decreased in anterior cingulate gyrus in ROI analysis (controls, 9 men and 6 women; p < 0.01). The area with significantly increased 11C-MPDX binding, lower frontal cortex, rolandic area, and posterior cingulate gyrus, did not overlap with the areas of neuronal loss detected by decreased 11C-FMZ binding and did not completely overlap with area of reduced18F-FDG uptake. We obtained the first 11C-MPDX PET images reflecting the A1R BPND in human DAI brain in vivo. 11C-MPDX depicted increased A1R BPND in the areas surrounding the injured brain, whereas 18F-FDG demonstrated reduction throughout the brain. The results suggested that A1R might continuously confer neuroprotective or neuromodulatory effects in DAI even in the chronic stage.

Изменение специфического связывания [N-метил-&lt;sup&gt;3&lt;/sup&gt;H]флунитразепама при воздействии анксиогенами

Изучено влияние лекарственных препаратов и фармакологических соединений, обладающих анксиогенными свойствами: йохимбин, кофеин, производное β-карболинов — соединение β-ССВ на специфическое связывание полного агониста бензодиазепинового участка ГАМКА рецептора [N-метил-3H]флунитразепама. Кофеин в дозе 100 мг/кг, n-бутил-β-карболин-3 карбоксилат (β-ССВ) в дозе 3 мг/кг и йохимбин в дозе 5 мг/кг вызывали анксиогенный эффект у мышей линии Balb/c и C57Bl/6. Анксиогенная активность кофеина, β-ССВ и йохимбина сопряжена с падением уровня бензодиазепиновой рецепции.

Altered SPECT (123)I-iomazenil Binding in the Cingulate Cortex of Children with Anorexia Nervosa.

Several lines of evidence suggest that anxiety plays a key role in the development and maintenance of anorexia nervosa (AN) in children. The purpose of this study was to examine cortical GABA(A)-benzodiazepine receptor binding before and after treatment in children beginning intensive AN treatment. Brain single-photon emission computed tomography (SPECT) measurements using 123I-iomazenil, which binds to GABA(A)-benzodiazepine receptors, was performed in 26 participants with AN who were enrolled in a multimodal treatment program. Sixteen of the 26 participants underwent a repeat SPECT scan immediately before discharge at conclusion of the intensive treatment program. Eating behavior and mood disturbances were assessed using Eating Attitudes Test with 26 items (EAT-26) and the short form of the Profile of Mood States (POMS). Clinical outcome scores were evaluated after a 1-year period. We examined association between relative iomazenil-binding activity in cortical regions of interest and psychometric profiles and determined which psychometric profiles show interaction effects with brain regions. Further, we determined if binding activity could predict clinical outcome and treatment changes. Higher EAT-26 scores were significantly associated with lower iomazenil-binding activity in the anterior and posterior cingulate cortex. Higher POMS subscale scores were significantly associated with lower iomazenil-binding activity in the left frontal, parietal cortex, and posterior cingulate cortex (PCC). “Depression–Dejection” and “Confusion” POMS subscale scores, and total POMS score showed interaction effects with brain regions in iomazenil-binding activity. Decreased binding in the anterior cingulate cortex and left parietal cortex was associated with poor clinical outcomes. Relative binding increases throughout the PCC and occipital gyrus were observed after weight gain in children with AN. These findings suggest that cortical GABAergic receptor binding is altered in children with AN. This may be a state-related change, which could be used to monitor and guide the treatment of eating disorders.

Differential effects of delayed aging on phenotype and striatal pathology in a murine model of Huntington disease.

The common neurodegenerative syndromes exhibit age-related incidence, and many Mendelian neurodegenerative diseases exhibit age-related penetrance. Mutations slowing aging retard age related pathologies. To assess whether delayed aging retards the effects of a mutant allele causing a Huntington's disease (HD)-like syndrome, we generated compound mutant mice, placing a dominant HD knock-in polyglutamine allele onto the slow-aging Snell dwarf genotype. The Snell genotype did not affect mutant huntingtin protein expression. Bigenic and control mice were evaluated prospectively from 10 to 100 weeks of age. Adult HD knock-in allele mice lost weight progressively with weight loss blunted significantly in male bigenic HD knock-in/Snell dwarf mice. Impaired balance beam performance developed significantly more slowly in bigenic HD knock-in/Snell dwarf mice. Striatal dopamine receptor expression was diminished significantly and similarly in all HD-like mice, regardless of the Snell genotype. Striatal neuronal intranuclear inclusion burden was similar between HD knock-in mice with and without the Snell genotype, whereas nigral neuropil aggregates were diminished in bigenic HD knock-in/Snell dwarf mice. Compared with control mice, Snell dwarf mice exhibited differences in regional benzodiazepine and cannabinoid receptor binding site expression. These results indicate that delaying aging delayed behavioral decline with little effect on the development of striatal pathology in this model of HD but may have altered synaptic pathology. These results indicate that mutations prolonging lifespan in mice delay onset of significant phenotypic features of this model and also demonstrate dissociation between striatal pathology and a commonly used behavioral measure of disease burden in HD models.

Findings of 123I-Iomazenil SPECT During and After Stroke-Like Episodes in a Patient With MELAS

Reduced cortical benzodiazepine receptor binding potential in late images of I-iomazenil SPECT indicates neuronal damage in the cortex. We present the case of a 31-year-old woman with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) who had focal seizures in the right hand. FLAIR imaging in the ictal state revealed a high-intensity lesion in the left post-central gyrus, while I-iomazenil SPECT showed decreased tracer uptake in this lesion. The lesion completely disappeared on FLAIR imaging performed 1 month after the focal seizures; in contrast, I-iomazenil SPECT still revealed a significant decrease in tracer uptake in this lesion.

Increased binding of peripheral benzodiazepine receptor in mild cognitive impairment–dementia converters measured by positron emission tomography with [11C]DAA1106

Subjects with mild cognitive impairment (MCI) have “prodromal or incipient” dementia with neuropathological changes. Peripheral benzodiazepine receptor (PBR) binding was shown to reflect activated microglia, one of the predictive biomarkers of conversion to dementia. We sought to evaluate PBR binding in MCI subjects using positron emission tomography (PET). PET scans with [11C]DAA1106, a potent and selective ligand for PBR, were performed on seven MCI subjects, 10 patients with Alzheimer's disease (AD) and 10 age-matched control subjects. PBR binding in the regions of interest was quantified by binding potential (BP). Five MCI subjects were clinically followed for 5years after their initial PET scans. [11C]DAA1106 binding to PBR was significantly increased in widespread areas in MCI subjects when compared to healthy controls. We found no significant difference in BP between MCI and AD patients. MCI subjects with [11C]DAA1106 binding values higher than the control mean +0.5 standard deviation (S.D.) developed dementia within 5years. Our finding of higher DAA binding in MCI subjects indicated that microglial activation may occur before the onset of dementia. In vivo detection of microglial activation may provide useful prognostic information with respect to stratifying MCI subjects at increased risk of dementia.

Synthesis, benzodiazepine receptor binding and molecular modelling of isochromeno[4,3-c]pyrazol-5(1H)-one derivatives

A series of isochromeno[4,3-c]pyrazole-5(1H)-one derivatives 7b–h were prepared and tested at 10μM for their ability to displace specific [3H]flunitrazepam from bovine brain membranes. The substitution pattern of the above derivatives was shown to influence the receptor affinity. The most active compound of the series was 7e, showing a 54% inhibition of [3H]flunitrazepam binding. Compounds 7a–d,i were compared with the known isomers chromeno[4,3-c]pyrazole-4(1H)-ones 14a–d,i, showing that the isochromene/chromene isomerism influences the activity.

Accuracy of Central Benzodiazepine Receptor Binding Potential/Cerebral Blood Flow SPECT Imaging for Detecting Misery Perfusion in Patients With Unilateral Major Cerebral Artery Occlusive Diseases

The aim of the present study was to determine whether central benzodiazepine receptor binding potential (BRBP)/cerebral blood flow (CBF) or a combination of CBF and cerebrovascular reactivity (CVR) to acetazolamide on single-photon emission computed tomography (SPECT) more accurately detects misery perfusion, indicating elevation of absolute value of oxygen extraction fraction (OEF) on positron emission tomography (PET), in patients with unilateral major cerebral artery occlusive diseases. In 84 patients, OEF, CBF, CVR to acetazolamide, and BRBP were assessed using ¹⁵O-PET and N-isopropyl-p-[¹²³I]-iodoamphetamine and [¹²³I]-iomazenil SPECT, respectively. A region of interest was automatically placed in the middle cerebral artery territory using a 3-dimensional stereotactic region of interest template. Sensitivity, specificity, and positive and negative predictive values for the affected side-to-contralateral side asymmetry on SPECT-BRBP/CBF to detect the abnormally elevated PET-OEF in the affected hemisphere were 100%, 86.4%, 66.7%, and 100%, respectively. Area under the receiver operating characteristic curve in detecting the abnormally elevated PET-OEF in the affected hemisphere did not differ between analysis of the combination of SPECT-CBF and SPECT-CVR in the affected hemisphere (0.89; 95% confidence interval, 0.80-0.94) and that of the affected side-to-contralateral side asymmetry on SPECT-BRBP/CBF (0.93; 95% confidence interval, 0.86-0.97). The combination of the 3 detected abnormally elevated PET-OEF with 97.0% specificity and 90.0% positive predictive value. The accuracy of central BRBP/CBF asymmetry on SPECT is equivalent to that of the combination of CBF and CVR to acetazolamide on SPECT for detecting misery perfusion in patients with unilateral major cerebral artery occlusive disease.

Combination of Preoperative Cerebral Blood Flow and 123I-Iomazenil SPECT Imaging Predicts Postoperative Cognitive Improvement in Patients Undergoing Uncomplicated Endarterectomy for Unilateral Carotid Stenosis

The purpose of the present study was to determine whether preoperative cerebral blood flow (CBF) or cortical central benzodiazepine receptor binding potential as measured by brain perfusion or (123)I-iomazenil (IMZ) single-photon emission computed tomography (SPECT) imaging, respectively, can predict cognitive improvement after uncomplicated carotid endarterectomy (CEA). Neuropsychological testing was performed preoperatively and after 1 month postoperatively in 140 patients who underwent CEA for unilateral internal carotid artery (ICA) stenosis (≥ 70%). Brain perfusion SPECT using N-isopropyl-p-[(123)I]-iodoamphetamine and IMZ SPECT was also performed before surgery. SPECT data were analyzed using a 3-dimensional stereotaxic region-of-interest template. Fourteen patients (10%) showed improved cognitive function postoperatively. Logistic regression analysis demonstrated that, among the variables tested, low relative CBF in the cerebral hemisphere ipsilateral to surgery was significantly associated with postoperative cognitive improvement (95% confidence interval: 0.623-0.868; P = 0.0003). The combination of reduced relative CBF and moderately reduced relative uptake of IMZ identified development of postoperative cognitive improvement with 100.0% sensitivity, 84.9% specificity, a positive predictive value of 42.4%, and a negative predictive value of 100%. Further, in a subgroup of patients with reduced relative CBF and moderately reduced relative uptake of IMZ, the difference between these 2 values was significantly greater in patients with postoperative cognitive improvement than in those without (P < 0.0001). The combination of preoperative CBF and cortical central benzodiazepine receptor binding potential as determined by SPECT can predict cognitive improvement after CEA.

CENTRAL NERVOUS SYSTEM DEPRESSANT ACTIVITY OF AQUEOUS EXTRACT OF CISSUS QUADRANGULARIS

The research work deals with the screening of aqueous extracts of Cissus quadrangularis (CQ) stem for central nervous system (CNS) depressant activity. The CQ leaves and stems contain resveratrol, piceatannol, pallidol perthenocissin derivatives as chemical components and are reported to have anticonvulant activity. The extract of plant showed GABA A - benzodiazepine receptor binding activity we made an attempt to study its CNS depressant effect. The different activities studied were test for locomotor activity, effect on muscle co-ordination, antiaggressive and antianxiety activities. The result of the study reflected that aqueous extract of the stem (150 mg/kg, p.o) decreased locomotor activity, produced muscle relaxation and showed antianxiety and antiaggressive activity

Preoperative Central Benzodiazepine Receptor Binding Potential and Cerebral Blood Flow Images on SPECT Predict Development of New Cerebral Ischemic Events and Cerebral Hyperperfusion After Carotid Endarterectomy

Risk factors for cerebrovascular complications developing during or after carotid endarterectomy (CEA) include preoperative impairments in cerebral hemodynamics, as detected by the demonstration of decreased cerebrovascular reactivity (CVR) to acetazolamide on brain perfusion SPECT. Central benzodiazepine receptor binding potential (CBRBP) and cerebral blood flow (CBF) images on SPECT provide high sensitivity and high specificity for detecting misery perfusion in patients with chronic unilateral major cerebral artery occlusive disease. The aim of the present study was to determine whether preoperative CBRBP/CBF images on SPECT could identify patients at risk for new cerebral ischemic events, including neurologic deficits and cerebral ischemic lesions on diffusion-weighted MRI, or cerebral hyperperfusion after CEA and to compare the predictive accuracy of CBRBP/CBF with that of CVR to acetazolamide on SPECT. CBF, CVR, and CBRBP were assessed using N-isopropyl-p-(123)I-iodoamphetamine ((123)I-IMP) and (123)I-iomazenil SPECT before CEA in 112 patients with unilateral internal carotid artery stenosis (≥ 70%). CBF measurement using (123)I-IMP SPECT was also performed immediately and 3 d after CEA. A region of interest was automatically placed in the middle cerebral artery territory in both cerebral hemispheres using a 3-dimensional stereotactic region-of-interest template. Diffusion-weighted MRI was performed within 3 d before and 24 h after surgery. A preoperative increase in the affected side-to-contralateral side asymmetry on CBRBP/CBF value was the only significant independent predictor of postoperative new cerebral ischemic events (95% confidence intervals [CI], 1.145-1.608; P = 0.0004) or post-CEA hyperperfusion (95% CI, 1.244-2.252; P = 0.0007). There was no difference in the ability to predict post-CEA hyperperfusion when comparing the area under the receiver-operating-characteristic curve of the affected side-to-contralateral side asymmetry on CBRBP/CBF and that of the CVR in the affected hemisphere. However, the former value (0.924; 95% CI, 0.854-0.972) was significantly greater than the latter value (0.782; 95% CI, 0.697-0.852) for the prediction of new postoperative cerebral ischemic events (P < 0.05). Preoperative CBRBP/CBF images on SPECT can more accurately identify patients at risk for cerebrovascular complications occurring during or after CEA when compared with preoperative CVR to acetazolamide.

Normal Phase HPLC as a Model System of Specific Benzodiazepine-receptor Binding

The hypothesis that benzodiazepines interact with their receptors in the CNS via hydrogen bonding mediated by the carbonyl and imine groups has been studied by correlating a physicochemical parameter affected by hydrogen bonds potentially accepted or donated by benzodiazepines with their binding constants to synaptosomal rat brain membranes. The reference physicochemical system chosen was normal phase high-performance liquid chromatography on mu-Bondapack NH2 (propylamine groups chemically bonded to porous silica) eluted with mixtures of n-hexane and 2-propanol. A strong correlation has been found for binding of the implicated groups to the column and to the receptor.

Central Benzodiazepine Receptor Binding Potential and CBF Images on SPECT Correlate with Oxygen Extraction Fraction Images on PET in the Cerebral Cortex with Unilateral Major Cerebral Artery Occlusive Disease

Oxygen extraction fraction (OEF) is a key predictor of stroke recurrence in patients with symptomatic major cerebral arterial occlusive disease. The purpose of the present study was to compare central benzodiazepine receptor binding potential (BRBP) and cerebral blood flow (CBF) images on SPECT with OEF images on PET in patients with chronic unilateral middle cerebral artery (MCA) or internal carotid artery (ICA) occlusive disease. OEF, CBF, and BRBP were assessed using (15)O PET and N-isopropyl-p-(123)I-iodoamphetamine and (123)I-iomazenil SPECT, respectively, in 20 healthy subjects and in 34 patients with unilateral MCA or ICA occlusive disease. All images were transformed into the standard brain size and shape by linear and nonlinear transformation using statistical parametric mapping for anatomic standardization. A region of interest (ROI) was automatically placed according to the arterial supply using a 3-dimensional stereotactic ROI template, and the ratio of the value in the affected side to that in the contralateral side was calculated in each image. Among patients with occlusive disease, a significant positive correlation was observed between PET OEF and SPECT BRBP/CBF ratios in 3 cerebral cortical regions (r = 0.851, P < 0.0001, for anterior cerebral artery [ACA] ROI; r = 0.807, P < 0.0001, for MCA ROI; and r = 0.774, P < 0.0001, for posterior cerebral artery [PCA] ROI), but there were no correlations between these 2 parameters in the basal ganglia or the cerebellum. When an abnormally elevated PET OEF ratio was defined as a value greater than the mean + 2 SDs obtained in healthy subjects, sensitivity and specificity were, respectively, 100% and 96% for the ACA ROI, 100% and 89% for the MCA ROI, and 100% and 93% for the PCA ROI for the SPECT BRBP/CBF ratio for detecting an abnormally elevated PET OEF ratio. BRBP/CBF images on SPECT correlate with OEF images on PET in a specific clinical setting-that is, in the cerebral cortex of patients with chronic unilateral MCA or ICA occlusive disease.

Modulation of benzodiazepine receptor, adrenoceptor and muscarinic receptor by diazepam in rat parotid gland

This study investigated the influence of diazepam on the binding characteristics of adrenoceptor, muscarinic and benzodiazepine receptors in rat parotid gland membrane using a radioligand binding assay. At a concentration of > 10 − 6 M, diazepam competed with [ 3H]dihydroalprenolol for β-adrenoceptor, but not [ 3H]prazosin for α-adrenoceptor or [ 3H]quinuclidinyl benzilate for muscarinic receptor. Continuous administration of diazepam at doses of 0.4 mg/kg/day, i.p. for 7 days in rat significantly decreased pilocarpine (4.0 mg/kg, i.p.)-induced parotid salivary flow. Diazepam also produced a significant increase in the dissociation constant (Kd) value for [ 3H]dihydroalprenolol binding, but no change in the maximal binding capacity (Bmax) value, and a decrease in the Kd value for [ 3H]diazepam binding to benzodiazepine receptors, but no change in the Kd or Bmax values for [ 3H]prazosin or [ 3H]quinuclidinyl benzilate binding. These results suggest that continuous administration of diazepam modifies affinity for β-adrenoceptor and benzodiazepine receptor binding sites in parotid gland membrane and that changes in these binding sites may be closely related to diazepam-induced suppression of salivary secretion.

Evaluation of the pharmacokinetic interaction of midazolam with ursodeoxycholic acid, ketoconazole and dexamethasone by brain benzodiazepine receptor occupancy

To clarify whether alterations in midazolam pharmacokinetics resulting from changes in cytochrome P450 3A (CYP3A) activity lead to changes in its pharmacodynamic effects, benzodiazepine receptor occupancy was measured in the brain of rats after oral administration of midazolam. Receptor occupancy was measured by radioligand binding assay in rats pretreated with ursodeoxycholic acid (UDCA), ketoconazole and dexamethasone, and the plasma concentration of midazolam was simultaneously determined. There was a significant increase in the apparent dissociation constant and decrease in the maximum number of binding sites for specific [(3) H]flunitrazepam binding after oral administration of midazolam at pharmacologically relevant doses, suggesting that midazolam binds significantly to brain benzodiazepine receptors. Pretreatment with UDCA significantly enhanced the binding. This correlated well with significant enhancement by UDCA of the plasma midazolam concentration. The brain benzodiazepine receptor binding of oral midazolam was significantly enhanced by pretreatment with ketoconazole, a potent inhibitor of CYP3A, whereas it was significantly reduced by treatment with dexamethasone, an inducer of this enzyme. These effects paralleled changes in the plasma concentration of midazolam. The results indicate that pharmacokinetic changes such as altered CYP3A activity significantly influence the pharmacodynamic effect of midazolam by affecting occupancy of benzodiazepine receptors in the brain. They also suggest in-vivo or ex-vivo time-dependent measurements of receptor occupancy by radioligand binding assay to be a tool for elucidating the pharmacokinetic interaction of benzodiazepines with other agents in pre-clinical and clinical evaluations.