Changes in cerebral metabolic activity in men undergoing androgen deprivation therapy for non-metastatic prostate cancer.

OBJECTIVE: Androgen deprivation therapy (ADT) is a common treatment option for men with biochemical relapse from prostate cancer. ADT is associated with changes in mood, cognition, and quality of life, and most recently with increased risk for Alzheimer's disease (AD). This study examined changes in brain metabolism using positron emission tomography (PET) in men undergoing intermittent ADT. METHODS: Nine men with prostate cancer and a rising PSA (biochemical recurrence) without evidence of metastases were treated with intermittent ADT consisting of 9 months of complete androgen blockade achieved with combined leuprolide acetate and flutamide. Patients underwent resting [Fuorine-18] fluorodeoxyglucose PET (18F-FDGPET) at baseline (before treatment) and again after 9 months of ADT. RESULTS: Whole-brain mapping analysis after 9 months of androgen deprivation compared to pretreatment baseline revealed decreased regional cerebral glucose metabolism in the cerebellum, posterior cingulate, and medial thalamus bilaterally. Associations of brain metabolism with measurements of cognition and mood while on androgen deprivation revealed positive correlations between the posterior cingulate, left inferior parietal lobule (BA40), and left mid temporal gyrus (BA39) and spatial reasoning and a negative correlation between left inferior parietal lobule and verbal memory. Several mood indices were negatively correlated with hypothalamus and brainstem. CONCLUSION: These findings suggest that complete androgen deprivation may result in changes in regional brain metabolism associated with variation in mood, verbal memory, and spatial performance. Brain regions that were impacted from ADT are similar and overlap with brain regions with metabolic decline found in early AD and diabetes, suggesting possible common mechanisms.

Morphological and parametric estimation of fetal neural stem cell migratory capacity in the rat brain.

Magnetic resonance imaging (MRI) can non-invasively monitor the migratory behavior of magnetically labeled stem cells after transplantation. Signal changes associated with the clearance of the contrast agent due to cell death and leaked tracer in the interstitial space must be better understood in order to accurately interpret imaging results. In this study, fetal neural stem cells were labeled with superparamagnetic iron oxide (SPIO) particles and transplanted into the corpus callosum of the adult rat. MRI was performed on the day of transplantation and at one week. Control subjects received injections of either non-viable, labeled cells or loose SPIO particles. Two quantitative image analysis algorithms were developed to evaluate imaging results: 1) signal intensity drop-out areas were segmented and compared on a pixel-wise basis between initial and one week images; and 2) signal intensity profiles of transplanted materials at one week were parametrically modeled to estimate migration speed. Segmentation results showed that the number of pixels segmented at one week was significantly greater than the initial number of segmented pixels for subjects receiving injections of viable cells as compared to controls (p<0.05). The average speed of migration of viable cells along the corpus callosum was 69.2+/-41.1 microm/d and was significantly higher than controls (p<0.05). This study demonstrates an in vivo assay to quantitatively evaluate stem cell migration that can be used in different experimental paradigms.

In vivo manganese MR imaging of calcium influx in spontaneous rat pituitary adenoma.

BACKGROUND AND PURPOSE: Rapid uptake of the calcium analog manganese (Mn2+) into spontaneous pituitary adenoma during MR imaging of aged rats generated the hypothesis that neuroendocrine tumors may have a corresponding increase in calcium influx required to trigger hormonal release. A goal of this study was to investigate the potential for clinical evaluation of pituitary adenoma by MR imaging combined with administration of Mn2+ (Mn-MR imaging). MATERIALS AND METHODS: Mn-MR imaging was used to characterize the dynamic calcium influx in normal aged rat pituitary gland as well as spontaneous pituitary adenoma. To confirm the validity of Mn2+ as a calcium analog, we inhibited Mn2+ uptake into the olfactory bulb and pituitary gland of normal rats by using the calcium channel blocker verapamil. Rats with adenomas received fluorodeoxyglucose-positron-emission tomography (FDG-PET) scanning for characterization of tumor metabolism. Mn2+ influx was characterized in cultured pituitary adenoma cells. RESULTS: Volume of interest analysis of the normal aged pituitary gland versus adenoma indicated faster and increased calcium influx in adenoma at 1, 3, 11, and 48 hours. Mn2+ uptake into the olfactory bulb and pituitary gland of normal rats was inhibited by calcium channel blockers and showed dose-dependent inhibition on dynamic MR imaging. FDG-PET indicated correlation between tumor energy metabolism and Mn2+ influx as well as tumor size. CONCLUSION: These results indicate that adenomas have increased activity-dependent calcium influx compared with normal aged pituitary glands, suggesting a potential for exploitation in the clinical work-up of pituitary and other neuroendocrine tumors by developing Mn-MR imaging for humans.

Magneto-optical labeling of fetal neural stem cells for in vivo MRI tracking.

Neural stem cell therapy for neurological pathologies, such as Alzheimer's and Parkinson's disease, may delay the onset of symptoms, replace damaged neurons and/or support the survival of endogenous cells. Magnetic resonance imaging (MRI) can be used to track magnetically labeled cells in vivo to observe migration. Prior to transplantation, labeled cells must be characterized to show that they retain their intrinsic properties, such as cell proliferation into neurospheres in a supplemented environment. In vivo images must also be correlated to sensitive, histological markers. In this study, we show that fetus-derived neural stem cells can be co-labeled with superparamagnetic iron oxide and PKH26, a fluorescent dye. Labeled cells retain the ability to proliferate into neurospheres in culture, but labeling prevents neurospheres from merging in a non-adherent culture environment. After labeled NSCs were transplantation into the rat brain, their location and subsequent migration along the corpus callosum was detected using MRI. This study demonstrates an imaging paradigm with which to develop an in vivo assay for quantitatively evaluating fetal neural stem cell migration.

A viral envelope as a vehicle for tracer, drug, and gene delivery: initial biodistribution study using PET imaging.

Viral envelopes can be used as an effective vehicle to deliver imaging tracers as well as therapeutic drugs and genes. However, the current methods for in vivo tracking of viral envelopes are limited. This purpose of this study is to investigate dynamically the in vivo biodistribution of viral envelopes using positron emission tomography (PET) imaging. The hemagglutinating virus of Japan envelope (HVJ-E) was labeled with radioactive fluorine (F-18) for tracking with PET imaging. Due to the low molecular weight of F-18, the encapsulation process by HVJ-E was optimized using the cationic agent poly-L-lysine (PLL, MW 66.7 kDa) and Feridex, a magnetic resonance imaging tracer. After labeling, HVJ-Es were injected intravenously into the normal rat and followed for 2 h using high resolution PET imaging. Region of interest analysis showed a significant increase in average liver accumulation based on radioactivity as compared to all control subjects. Average brain uptake showed a significant increase in radioactivity as compared to control subjects receiving F-18-PLL complexes or F-18 alone. Control subjects showed F-18 uptake primarily in the bones. These results demonstrate a molecular imaging technique that can be used to monitor drug and gene delivery and evaluate potential targeting mechanisms.

A Viral Envelope as a Vehicle for Tracer, Drug, and Gene Delivery: Initial Biodistribution Study Using PET Imaging.

Viral envelopes can be used as an effective vehicle to deliver imaging tracers as well as therapeutic drugs and genes. However, the current methods for in vivo tracking of viral envelopes are limited. This purpose of this study is to investigate dynamically the in vivo biodistribution of viral envelopes using positron emission tomography (PET) imaging. The hemagglutinating virus of Japan envelope (HVJ-E) was labeled with radioactive fluorine (F-18) for tracking with PET imaging. Due to the low molecular weight of F-18, the encapsulation process by HVJ-E was optimized using the cationic agent poly-L-lysine (PLL, MW 66.7 kDa) and Feridex, a magnetic resonance imaging tracer. After labeling, HVJ-Es were injected intravenously into the normal rat and followed for 2 h using high resolution PET imaging. Region of interest analysis showed a significant increase in average liver accumulation based on radioactivity as compared to all control subjects. Average brain uptake showed a significant increase in radioactivity as compared to control subjects receiving F-18-PLL complexes or F-18 alone. Control subjects showed F-18 uptake primarily in the bones. These results demonstrate a molecular imaging technique that can be used to monitor drug and gene delivery and evaluate potential targeting mechanisms.

Sprouting and synaptic reorganization in the subiculum and CA1 region of the hippocampus in acute and chronic models of partial-onset epilepsy.

Repeated seizures induce permanent alterations in the hippocampal circuitry in experimental models and patients with intractable temporal lobe epilepsy (TLE). Most studies have concentrated their attention on seizure-induced reorganization of the mossy fiber pathway. The present study examined the projection pathway of the CA1 pyramidal neurons to the subiculum, which is the output of the hippocampal formation in five models of TLE. We examined the laminar pattern of Timm's histochemistry in the stratum lacunosum-moleculare of CA1 in three acute and two chronic models of TLE: intraventricular kainic acid (KA), systemic KA, systemic pilocarpine, chronic electric kindling and chronic i.p. pentylenetetrazol. The laminar pattern of Timm histochemistry in the stratum moleculare of CA1 was permanently remodeled in epileptic models suggesting sprouting of Timm containing terminals from the adjacent stratum lacunosum. Ultrastructural examination confirmed that Timm granules were localized in synaptic terminals. As the source of Timm-labeled terminals in this region was not known, sodium selenite, a selective retrograde tracer for zinc-containing terminals, was iontophoretically injected in vivo in rats exposed to systemic pilocarpine, systemic KA or chronic pentylenetetrazol. The normal projection of CA1 pyramidal neurons to the subiculum is topographically organized in a lamellar fashion. In normal rats, the extent of the injection site (terminals) and the retrogradely labeled pyramidal neurons (cell soma) corresponded to the same number of lamellas. In epileptic rats, the retrograde labeling extended 42-67% farther than the normal dorso-ventral extent including lamellas above and below the expected. This is direct evidence for sprouting of CA1 pyramidal axons into the subiculum and stratum lacunosum-moleculare of the CA1 region confirming the alterations of the laminar pattern of Timm's histochemistry. Sprouting of the CA1 projection to subiculum across hippocampal lamellas might lead to translamellar hyperexcitability, and to amplification and synchronization of epileptic discharges in the output gate of the hippocampal formation.

Efficiency of transfection and localization of superparamagnetic iron oxide particles in neural progenitor cells using two methods.

Stem cells represent a potentially revolutionary therapy for neurological pathologies but for which a thorough investigation of cell behavior in the living nervous system has yet to be performed. Contrast-enhanced cell tracking with magnetic resonance imaging can enable this investigation by introducing superparagmagnetic iron oxide (SPIO) particles within the cell membrane. Before magnetically labeled cells can be observed in vivo, it is essential to maximize SPIO transfer into the cell and to fully understand the localization of the contrast agent in mature neural cells. For practical applications, a quantitative evaluation of labeled cells before implantation will allow in vivo assertions. In this study, we present a comparison between two methods for magnetic transfection of neural progenitor cells: the hemmaglutinating virus of Japan envelope (HVJ-E) as a viral vector and a liposomal reagent. We show that HVJ-E is a more efficient vehicle of cell transfection using quantitative evaluation and that the iron content per cell can be predicted using a simple, automated image analysis of stained, labeled cells. Image analysis is also used in this study to show that the contrast agent is distributed in the axon after differentiation, an important aspect of understanding cell tracking in vivo.

Statistical brain mapping of 18F-FDG PET in Alzheimer's disease: validation of anatomic standardization for atrophied brains.

UNLABELLED: Despite the increased use of statistical mapping to detect brain functional changes in Alzheimer's disease (AD), potential artifacts introduced by stereotactic anatomic standardization of atrophied brains have not been examined carefully. We investigated the effects of anatomic standardization by Statistical Parametric Mapping (SPM) and NEUROSTAT. METHODS: First, 10 AD patients and 10 age-matched healthy volunteers underwent 18F-FDG brain PET imaging. Each image set was standardized to a stereotactic brain template using SPM or NEUROSTAT, followed by pixel normalization to the global or cerebellar activity. Within-group comparisons of standardized image sets by each method and a between-group comparison of healthy volunteers and AD patients were performed using the statistical analysis routines of SPM. Second, simulated PET image sets were generated from segmented MR image sets of 5 healthy volunteers and 5 AD patients. Using the anatomic standardization parameters estimated on the simulated image sets, original gray matter MR image sets were transformed to the stereotactic coordinate system. Between-group subtraction analyses of the transformed gray matter image sets between healthy volunteers and AD groups were performed to examine the accuracy of cortical gray matter matching. RESULTS: Between-group comparison by SPM or NEUROSTAT showed generally similar areas of hypometabolism in bilateral temporoparietal, posterior cingulate, and left frontal cortices. Both methods showed possible deformation artifacts in the anterior part of the corpus callosum. The localization of the peak hypometabolism varied considerably between the two methods when global normalization was applied. The use of a common brain template for standardization resulted in asymmetric differences in cortical margins, indicating systematic differences in the deformation algorithms. The realistic simulation study revealed gray matter mismatches to be 20% greater with SPM than with NEUROSTAT. CONCLUSION: Although different statistical mapping methods may yield grossly similar patterns of hypometabolism in AD, the extent, severity, and peak location of metabolic changes can be inconsistent. Deformation accuracy appears to be more prone to atrophy. These limitations need to be considered carefully in the application and interpretation of brain mapping analysis in atrophied brains.

Effects of nicotine on regional cerebral glucose metabolism in awake resting tobacco smokers.

Eleven healthy tobacco smoking adult male volunteers of mixed race were tobacco abstinent overnight for this study. In each subject, positron emission tomographic images of regional cerebral metabolism of glucose with [18F]fluorodeoxyglucose were obtained in two conditions in the morning on different days: about 3min after approximately 1-2mg of nasal nicotine spray and after an equivalent volume of an active placebo spray of oleoresin of pepper in a random counterbalanced design. A Siemens/CTI 931/08-12 scanner with the capability of 15 horizontal brain slices was used. The images were further converted into a standard uniform brain format in which the mean data of all 11 subjects were obtained. Images were analysed in stereotactic coordinates using pixel-wise t statistics and a smoothed Gaussian model. Peak plasma nicotine levels varied three-fold and the areas under the curve(0-30min) varied seven-fold among the individual subjects. Nicotine caused a small overall reduction in global cerebral metabolism of glucose but, when the data were normalized, several brain regions showed relative increases in activity. Cerebral structures specifically activated by nicotine (nicotine minus pepper, Z score >4.0) included: left inferior frontal gyrus, left posterior cingulate gyrus and right thalamus. The visual cortex, including the right and left cuneus and left lateral occipito-temporal gyrus fusiformis, also showed an increase in regional cerebral metabolism of glucose with Z scores >3. 6. Structures with a decrease in regional cerebral metabolism of glucose (pepper minus nicotine) were the left insula and right inferior occipital gyrus, with Z scores >3.5. Especially important is the fact that the thalamus is activated by nicotine. This is consistent with the high density of nicotinic cholinoceptors in that brain region. However, not all brain regions affected by nicotine are known to have many nicotinic cholinoceptors. The results are discussed in relation to the cognitive effects of nicotine.

Three-dimensional stereotactic surface projection analysis of macaque brain PET: development and initial applications.

UNLABELLED: To characterize better the local brain functions of conscious rhesus macaques, we developed automated image analysis techniques for monkey PET images, examined the cerebral glucose metabolism of monkeys, and compared it with that of humans. METHODS: Glucose metabolic PET images from 11 monkeys were obtained using a high-resolution animal PET scanner after intravenous administration of FDG. T1-weighted MR images were obtained from 6 of the monkeys. Referencing a bicommissural stereotactic macaque brain atlas, we created a PET brain template using coregistered MR images. Each individual PET image set was transformed to the PET template through an automated affine transformation, followed by nonlinear warping along the directions of the major neuronal fiber bundles in the brain. For minimization of residual anatomic variability, metabolic activities were extracted using 3-dimensional stereotactic surface projections. The effects of anatomic standardization were evaluated using MR images. Patterns of cerebral glucose metabolism of young versus aged monkeys were examined. The metabolic activities of aged monkeys were compared with those of elderly healthy human volunteers that had been analyzed similarly. RESULTS: Anatomic standardization reduced individuals' anatomic variability as evidenced by a reduction in the number of MR pixels with higher SDs calculated across monkeys. Coefficient-of-variation maps of conscious monkeys revealed that the greatest metabolic variances were near the central sulci and occipital cortices. Age-associated glucose metabolic reductions were most pronounced in the occipital lobe, caudate nucleus, and temporal lobe. Compared with human brains, the monkey frontal lobe and posterior cingulate gyrus had significantly less metabolic activity and the supramarginal gyrus and vermis had significantly more metabolic activity. CONCLUSION: The proposed method permits pixel-by-pixel characterization of the metabolic activities of rhesus macaque brains in the stereotactic coordinate system. Greater metabolic variances in the central sulcus region and occipital lobe suggest potential difficulties in controlling sensory input and motor output or planning in conscious monkey experiments. The analyses revealed age-related metabolic reductions in monkeys and marked differences in metabolic patterns between aged monkey brains and aged human brains. The proposed brain-mapping technique enables reproducible and observer-independent analyses and will serve as an important investigative tool for primate brain imaging research.

Beta(2)-adrenoceptor polymorphism determines vascular reactivity in humans.

Altered beta-adrenergic regulation has been reported in individuals with hypertension. The variability in vascular responsiveness to beta-agonists, such as isoproterenol, observed in humans may be explained partially by beta(2)-adrenoceptor polymorphism. Individuals with the Gln27 form of the receptor may show reduced vascular reactivity because of downregulation expression of the receptor in the vasculature. We screened 127 normotensive white subjects, 37 of whom were homozygous for these alleles. Thirty-two subjects (17 Gln27 and 15 Glu27) agreed to receive brachial artery infusions of isoproterenol at doses of 1 to 300 ng. min(-1); forearm blood flow was measured by using venous occlusion plethysmography. Of these subjects, 25 (12 Glu27 and 13 Gln27) received local doses of isoproterenol (0.3 to 30.0 ng. min(-1)) via a dorsal hand vein preconstricted with norepinephrine. Compared with subjects homozygous for the Glu27 allele, subjects with the Gln27 substitution had lower baseline blood flow and, in response to isoproterenol, had a significantly attenuated increase in forearm blood flow. This pattern was more marked in veins. We also studied the relationship between the position 16 polymorphism and vascular reactivity. Homozygotes for Arg16 had significantly lower basal blood flow and attenuated increases in forearm blood flow compared with the Gly16 homozygotes. This was significant in veins but not in arteries. Thus, beta(2)-adrenoceptor genotype determines vascular responses to isoproterenol in forearm resistance vessels and in capacitance vessels. Further studies are necessary to establish whether beta(2)-adrenoceptor polymorphisms are important in the genesis of hypertension.

Discordance between traditional pathologic and energy metabolic changes in very early Alzheimer's disease. Pathophysiological implications.

These results suggest that neither the loss of entorhinal efferents nor cholinergic deficit explains all the metabolic features seen in very early AD. Given recent immunohistological evidence of massive glutamatergic synaptic alteration in early AD cortex and insights into neuronal and glial mechanisms of glucose metabolism, very early metabolic changes in AD probably reflect a significant impairment of glycolytic activities in the cortico-cortical glutamatergic systems in a preclinical stage of the disease. However, the exact mechanisms of such impairment in these neurons are yet to be determined.

The role of lignin as a defence against the spruce bark beetle Dendroctonus micans: effect on larvae and adults.

The role of lignin as a physical defence against Dendroctonus micans was investigated in laboratory feeding experiments. The effect of lignin is dose-dependent, reducing larval survival, growth rate, and weight, as well as affecting gallery construction. Adults lay fewer eggs in lignified bark and also tend to construct abnormal galleries. The distribution of lignin in trees suggests a role in defence against bark beetles that feed in the thicker bark on the lower bole.