Autoradiographic Techniques Research Articles

Correlation Between Severity of Carotid Stenosis and Vascular Reserve Measured by Acetazolamide Brain Perfusion Single Photon Emission Computed Tomography

Few studies have investigated the relationship between the degree of stenosis of the internal carotid artery (ICA) and cerebrovascular reserve (CVR). This study examined that relationship. A total of 56 ICAs in 43 patients were included. Computed tomography scan or magnetic resonance imaging showed no evidence of infarction in any of these patients. Both iodine-123-N-isopropyl-p-iodoamphetamine ((123)IMP)-single photon emission computed tomography (SPECT) in the resting state and (123)IMP-SPECT with acetazolamide (ACZ) enhancement were performed. Quantitated cerebral blood flow (CBF) images were acquired with the (123)IMP autoradiography technique. The mean CBF without ACZ administration (resting CBF) and CVR in the middle cerebral artery territory were calculated using stereotactic extraction estimation (SEE) analysis software. The degree of stenosis in the origin of the ICA was calculated from intra-arterial digital subtraction angiography. Resting CBF was not correlated with the degree of ICA stenosis; however, nonlinear regression analysis (second-order equation) showed a moderate correlation between CVR and the degree of ICA stenosis. In 72% of the cases with a CVR <30%, ICA stenosis was >74%. Using the SEE method, CVR was moderately correlated with the degree of ICA stenosis. Our findings indicate that evaluating CVR by ACZ-enhanced (123)IMP-SPECT in patients with ICA stenosis is of clinical value.

15 years of in situ cement–argillite interaction from Tournemire URL: Characterisation of the multi-scale spatial heterogeneities of pore space evolution

The solution selected by some countries to isolate radioactive wastes from the biosphere for up to one million years in deep geological repositories includes a multi-barrier disposal design, with steel canister, bentonite and cement materials. The geochemical contrast between such materials and the host rock formation creates perturbations potentially altering the confinement properties of the formation. In this context, the French Institute for the Radiological protection and Nuclear Safety (IRSN) have developed an in situ experimental programme based on the study of cement/argillaceous formation interfaces in their Underground Research Laboratory at Tournemire (Aveyron, France). An in situ engineered analogue of a cement/clay-rock interface which has undergone 15 years of interaction has been characterised. Such important interaction time for an in situ engineered analogue provides a bridge between laboratory-derived data and the long time scale of safety assessment modelling. As the mineralogical and petrological investigations have already been published, this work presents for the first time a quantitative characterisation of the spatial distribution of the porosity in the cement and the clay-rock in terms of time scale and design. Interfaces have been characterised using an autoradiography technique in addition to petrophysical measurements. This technique enables visualisation and quantification of the spatial distribution of the porosity using 2D mapping of decimetric-scale specimens. Thus autoradiographs allow highlighting the relationship between the field heterogeneities and the pore space evolution in each material in contact. Moreover, the porosity measurements show a clogging of the porosity in the clay-rock while the porosity increases in the cement. The extension of the porosity evolution extends to a centimetre on both sides of the interface but is heterogeneously distributed in space as a function of the fissure network and interface geometries. The connected fissure network visualised using autoradiography in the clogged area could permit solute (e.g. radionuclide) transport and may also be interpreted as an evolution of the mechanical properties of the clay-rock formation upon alkaline perturbation. This set of data, with the spatial quantification of the porosity in both cement and clay materials will be useful to constrain reactive transport modelling and thus to predict long term evolution of an engineered barrier.

Statistical parametric maps of 18F-FDG PET and 3-D autoradiography in the rat brain: a cross-validation study

Although specific positron emission tomography (PET) scanners have been developed for small animals, spatial resolution remains one of the most critical technical limitations, particularly in the evaluation of the rodent brain. The purpose of the present study was to examine the reliability of voxel-based statistical analysis (Statistical Parametric Mapping, SPM) applied to (18)F-fluorodeoxyglucose (FDG) PET images of the rat brain, acquired on a small animal PET not specifically designed for rodents. The gold standard for the validation of the PET results was the autoradiography of the same animals acquired under the same physiological conditions, reconstructed as a 3-D volume and analysed using SPM. Eleven rats were studied under two different conditions: conscious or under inhalatory anaesthesia during (18)F-FDG uptake. All animals were studied in vivo under both conditions in a dedicated small animal Philips MOSAIC PET scanner and magnetic resonance images were obtained for subsequent spatial processing. Then, rats were randomly assigned to a conscious or anaesthetized group for postmortem autoradiography, and slices from each animal were aligned and stacked to create a 3-D autoradiographic volume. Finally, differences in (18)F-FDG uptake between conscious and anaesthetized states were assessed from PET and autoradiography data by SPM analysis and results were compared. SPM results of PET and 3-D autoradiography are in good agreement and led to the detection of consistent cortical differences between the conscious and anaesthetized groups, particularly in the bilateral somatosensory cortices. However, SPM analysis of 3-D autoradiography also highlighted differences in the thalamus that were not detected with PET. This study demonstrates that any difference detected with SPM analysis of MOSAIC PET images of rat brain is detected also by the gold standard autoradiographic technique, confirming that this methodology provides reliable results, although partial volume effects might make it difficult to detect slight differences in small regions.

Transfer of 14C-photosynthate to the sporocarp of an ectomycorrhizal fungus Laccaria amethystina

Sporocarps of ectomycorrhizal fungi are strong carbon sinks for the source in host trees, but the details of carbon transfer from the host to the sporocarp are unknown. In this study, single seedlings of Japanese red pine (Pinus densiflora) colonised by Laccaria amethystina were grown on floral foam plates fitted in rhizoboxes, resulting in fruiting on the substrate. The seedlings were photosynthetically labelled with (14)CO(2); (14)C-labelled photosynthate transfer from leaves to sporocarps was then chased using a time-course autoradiography technique. (14)C was transferred to healthy, fresh sporocarps in a purple colour ranging from primordial to elongate sporocarps, but hardly to senesced ones that had faded to white or grey, or browned. This suggested that C is transferred only to physiologically active sporocarps. Two seedlings associated with a growing sporocarp were labelled again 7 and 16 days after the first labelling, respectively. (14)C accumulation in the sporocarps rose in a stepwise manner after the second labelling, indicating that sporocarps mainly used recently rather than previously photosynthesised C.

Examination of the translocation of sulfonylurea herbicides in sunflower plants by matrix‐assisted laser desorption/ionisation mass spectrometry imaging

Pesticides are widely used in agriculture to control weeds, pests and diseases. Successful control is dependent on the compound reaching the target site within the organism after spray or soil application. Conventional methods for determining uptake and movement of herbicides and pesticides include autoradiography, liquid scintillation and chromatographic techniques such as high-performance liquid chromatography (HPLC). Autoradiography using radiolabelled compounds provides the best indication of a compound's movement within the plant system. Autoradiography is an established technique but it relies on the synthesis of radiolabelled compounds. The distribution of four sulfonylurea herbicides in sunflower plants has been studied 24 h after foliar application. The use of matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) images of protonated molecules and fragment ions (resulting from fragmentation at the urea bond within the sulfonylurea herbicides) has provided evidence for translocation above and below the application point. The translocation of nicosulfuron and azoxystrobin within the same plant system has also been demonstrated following their application to the plant stem. This study provides evidence that MALDI-MSI has great potential as an analytical technique to detect and assess the foliar, root and stem uptake of agrochemicals, and to reveal their distribution through the plant once absorbed and translocated.

The α-Camera: A Quantitative Digital Autoradiography Technique Using a Charge-Coupled Device for Ex Vivo High-Resolution Bioimaging of α-Particles

Bioconjugates used in internal radiotherapy exhibit heterogeneous distributions in organs and tumors, implying a risk of nonuniform dose distribution in therapeutic applications using α-particle emitters. Tools are required that provide data on the activity distribution for estimation of absorbed dose on a suborgan level. The α-camera is a quantitative imaging technique developed to detect α-particles in tissues ex vivo. The aim of this study was to evaluate the characteristics of this imaging system and to exemplify its potential use in the development of α-radioimmunotherapy. The α-camera combines autoradiography with a scintillating technique and optical registration by a charge-coupled device (CCD). The imaging system characteristics were evaluated by measurements of linearity, uniformity, and spatial resolution. The technique was applied for quantitative imaging of (211)At activity distribution in cryosections of tumors, kidney, and whole body. Intratumoral activity distributions of tumor-specific (211)At-MX35-F(ab')(2) were studied at various times after injection. The postinjection activity distributions in the renal cortex and whole kidneys were compared for (211)At-F(ab')(2) and (211)At-IgG trastuzumab. Quantitative analysis of α-camera images demonstrated that the pixel intensity increased linearly with activity in the imaged specimen. The spatial resolution was 35 ± 11 μm (mean ± SD) and the uniformity better than 2%. Kidney cryosections revealed a higher cortex-to-whole kidney ratio for (211)At-F(ab')(2) than for (211)At-IgG (1.38 ± 0.03 and 0.77 ± 0.04, respectively) at 2 h after injection. Nonuniform intratumoral activity distributions were found for tumor-specific (211)At-MX35-F(ab')(2) at 10 min and 7 h after injection; after 21 h, the distribution was more uniform. The characteristics of the α-camera are promising, suggesting that this bioimaging system can assist the development, evaluation, and refinement of future targeted radiotherapy approaches using α-emitters. The α-camera provides quantitative data on the activity distribution in tissues on a near-cellular scale and can therefore be used for small-scale dosimetry, improving the prediction of biologic outcomes with α-particles with short path length and high linear energy transfer.

P.1.d.009 Longterm memory formation in non-association learning task in sham-operated and dentate gyrus lesioned rats

Previous studies have indicated that antidepressant drugs displaying different pharmacological profiles, administered repeatedly, increase the locomotor hyperactivity induced by various dopaminomimetics, among others by quinpirole. As this drug, according to a recent study, shows high affinity not only for dopamine D2 but also for dopamine D3 receptors, the question arises if dopamine D3 receptors are involved in the increase in quinpirole-elicited locomotor hyperactivity induced by repeated treatment with antidepressant drugs. In the present study we administered imipramine, amitriptyline, citalopram and mianserin (in a dose of 10 mg/kg p.o., twice a day, 14 days) to male Wistar rats and then (±)-7-OH-DPAT (7-hydroxy-dipropylaminotetralin), a dopamine D3 receptor agonist, was given (3 mg/kg s.c.). Hyperlocomotion induced by (±)-7-OH-DPAT was significantly increased by repeated administration of antidepressant drugs. The receptor autoradiography technique with [3H]7-OH-DPAT as a radioligand was applied to measure the effects of antidepressant drugs treatment on the dopamine D3 receptors in the islands of Calleja and in the shell of the nucleus accumbens septi, which are brain regions with highly selective expression of dopamine D3 receptors. The biochemical studies indicated that in both examined brain regions there was an increase in the binding of [3H]7-OH-DPAT following the repeated administration of antidepressant drugs. In some cases this increase was also observed after the acute administration of antidepressants. The results obtained in the present study indicate that antidepressant drugs administered repeatedly enhance the responsiveness of dopamine D3 receptors, probably via an increase in the density of these receptors. This mechanism is probably similar to that observed already in the case of dopamine D2 receptors. Therefore it is hypothesized that dopamine D3 receptors are also involved in the increased responsiveness to dopamine D3 receptor agonists observed after antidepressants.

Conversation with Michael Kuhar

Conversation with Michael Kuhar

Analytical Methods in Diffusion Studies

Research Article| January 01, 2010 Analytical Methods in Diffusion Studies Daniele J. Cherniak; Daniele J. Cherniak Department of Earth and Environmental Sciences Rensselaer Polytechnic Institute, chernd@rpi.edu Search for other works by this author on: GSW Google Scholar Richard Hervig; Richard Hervig School of Earth and Space Exploration, Arizona State University Search for other works by this author on: GSW Google Scholar Jürgen Koepke; Jürgen Koepke Institut für Mineralogie, Leibniz Universitaet Hannover Search for other works by this author on: GSW Google Scholar Youxue Zhang; Youxue Zhang Department of Geological Sciences, University of Michigan Search for other works by this author on: GSW Google Scholar Donggao Zhao Donggao Zhao Department of Geological Sciences, University of Texas at Austin Search for other works by this author on: GSW Google Scholar Author and Article Information Daniele J. Cherniak Department of Earth and Environmental Sciences Rensselaer Polytechnic Institute, chernd@rpi.edu Richard Hervig School of Earth and Space Exploration, Arizona State University Jürgen Koepke Institut für Mineralogie, Leibniz Universitaet Hannover Youxue Zhang Department of Geological Sciences, University of Michigan Donggao Zhao Department of Geological Sciences, University of Texas at Austin Publisher: Mineralogical Society of America First Online: 09 Mar 2017 © The Mineralogical Society Of America Reviews in Mineralogy and Geochemistry (2010) 72 (1): 107–170. https://doi.org/10.2138/rmg.2010.72.4 Article history First Online: 09 Mar 2017 Cite View This Citation Add to Citation Manager Share Icon Share Facebook Twitter LinkedIn MailTo Tools Icon Tools Get Permissions Search Site Citation Daniele J. Cherniak, Richard Hervig, Jürgen Koepke, Youxue Zhang, Donggao Zhao; Analytical Methods in Diffusion Studies. Reviews in Mineralogy and Geochemistry 2010;; 72 (1): 107–170. doi: https://doi.org/10.2138/rmg.2010.72.4 Download citation file: Ris (Zotero) Refmanager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentBy SocietyReviews in Mineralogy and Geochemistry Search Advanced Search In this chapter, we provide an overview of analytical methods used in diffusion studies. These methods fall into two broad categories – direct profiling and bulk release/exchange. Direct profiling methods, where the concentration of diffusant vs. depth is determined, can employ a variety of techniques, and can access diffusivities ranging from ~10−9 to 10−24 m2/s. Among these methods are the “classical” techniques of serial sectioning and autoradiography, which, despite a long history of application in diffusion studies, are now rarely used, having been superseded by other analytical methods with better depth resolution and greater flexibility. Direct... You do not have access to this content, please speak to your institutional administrator if you feel you should have access.

Subarachnoid hemorrhage induces enhanced expression of thromboxane A 2 receptors in rat cerebral arteries

Cerebral ischemia remains the key cause of morbidity and mortality after subarachnoid hemorrhage (SAH) with a pathogenesis that is still poorly understood. The aim of the present study was to examine the involvement of thromboxane A 2 receptors (TP) in the pathophysiology of cerebral ischemia after SAH in cerebral arteries. SAH was induced in rats by injecting 250 μl of blood into the prechiasmatic cistern. Two days after the SAH, cerebral arteries were harvested and contractile responses to the TP receptor agonist U46619 were investigated with myographs. In addition, the contractile responses were examined after pretreatment with selective TP receptor antagonist GR3219b. The TP receptor RNA and protein levels were analyzed by quantitative real-time PCR and immunohistochemistry, respectively. The global and regional cerebral blood flows (CBFs) were quantified with an autoradiographic technique. SAH resulted in enhanced contractile responses to U46619 as compared to sham. The TP receptor antagonist GR3219b abolished the enhanced contractile responses to U46619 observed after SAH. The TP receptor mRNA level was elevated after SAH as compared to sham. The level of TP receptor protein on the smooth muscle cells (SMCs) was increased in SAH compared to sham. Global and regional CBFs were reduced in SAH as compared to sham. The results demonstrate that SAH results in CBF reduction and this is associated with the enhanced expression of TP receptors in the SMC of cerebral arteries and microvessels.

Acute tryptophan depletion potentiates 3,4‐methylenedioxymethamphetamine‐induced cerebrovascular hyperperfusion in adult male wistar rats

The serotonergic (5-hydroxytryptamine; 5-HT) dysfunction found in depression may affect not only brain function (mood) but also cerebrovascular control. Similar, but possibly occult, disturbances may also be induced by 3,4-methylenedioxymethamphetamine-induced neurotoxicity (MDMA, or "ecstasy"). Acute tryptophan depletion (ATD) is widely used to identify vulnerability to depression, and we hypothesized that repeated MDMA administration would increase the sensitivity of rats to this acute serotonergic challenge. In this study, male Wistar rats were injected with MDMA (20 mg kg(-1), twice daily for 4 days) and challenged 3 weeks later with ATD, induced by intragastric administration of a nutritional mixture with tryptophan (TRP) removed. Cerebral metabolism (CMRG) and blood flow (CBF) were measured in parallel groups of animals following ATD by using quantitative [(14)C]2-deoxyglucose and [(14)C]iodoantipyrine autoradiographic techniques, respectively. A significant reduction in paroxetine binding to 5-HT transporter sites in MDMA-treated rats indicated 5HT terminal depletion, whereas the plasma TRP/sum large neutral amino acids ratio was reduced by 40% following ATD. Under all experimental conditions, the normal close correlation between CBF and metabolic demand was maintained. However, a global analysis of all brain regions revealed a significant decrease in the overall ratio of CBF to CMRG after ATD in control animals, whereas a higher ratio was observed after ATD in the MDMA-treated group. This increase in blood flow relative to cerebral metabolism suggests an ATD-induced loss of cerebrovascular tone in MDMA-treated animals that could have pathophysiological consequences and might conceivably contribute to the behavioral dysfunction of depression.

Performance assessment of the BEBIG MultiSource® high dose rate brachytherapy treatment unit

A comprehensive system characterisation was performed of the Eckert & Ziegler BEBIG GmbH MultiSource® High Dose Rate (HDR) brachytherapy treatment unit with an 192Ir source. The unit is relatively new to the UK market, with the first installation in the country having been made in the summer of 2009. A detailed commissioning programme was devised and is reported including checks of the fundamental parameters of source positioning, dwell timing, transit doses and absolute dosimetry of the source. Well chamber measurements, autoradiography and video camera analysis techniques were all employed. The absolute dosimetry was verified by the National Physical Laboratory, UK, and compared to a measurement based on a calibration from PTB, Germany, and the supplied source certificate, as well as an independent assessment by a visiting UK centre. The use of the ‘Krieger’ dosimetry phantom has also been evaluated. Users of the BEBIG HDR system should take care to avoid any significant bend in the transfer tube, as this will lead to positioning errors of the source, of up to 1.0 mm for slight bends, 2.0 mm for moderate bends and 5.0 mm for extreme curvature (depending on applicators and transfer tube used) for the situations reported in this study. The reason for these errors and the potential clinical impact are discussed. Users should also note the methodology employed by the system for correction of transit doses, and that no correction is made for the initial and final transit doses. The results of this investigation found that the uncorrected transit doses lead to small errors in the delivered dose at the first dwell position, of up to 2.5 cGy at 2 cm (5.6 cGy at 1 cm) from a 10 Ci source, but the transit dose correction for other dwells was accurate within 0.2 cGy. The unit has been mechanically reliable, and source positioning accuracy and dwell timing have been reproducible, with overall performance similar to other existing HDR equipment. The unit is capable of high quality brachytherapy treatment delivery, taking the above factors into account.

Equal contribution of increased intracranial pressure and subarachnoid blood to cerebral blood flow reduction and receptor upregulation after subarachnoid hemorrhage

Cerebral ischemia remains the key cause of disability and death in the late phase after subarachnoid hemorrhage (SAH), and its pathogenesis is still poorly understood. The purpose of this study was to examine whether the change in intracranial pressure or the extravasated blood causes the late cerebral ischemia and the upregulation of receptors or the cerebral vasoconstriction observed following SAH. Rats were allocated to 1 of 3 experimental conditions: 1) cisternal injection of 250 microl blood (SAH Group), 2) cisternal injection of 250 microl NaCl (Saline Group), or 3) the same procedure but without fluid injection (Sham Group). Two days after the procedure, the basilar and middle cerebral arteries were harvested, and contractile responses to endothelin (ET)-1 and 5-carboxamidotryptamine (5-CT) were investigated by means of myography. In addition, real-time polymerase chain reaction was used to determine the mRNA levels for ET(A), ET(B), and 5-HT(1) receptors. Regional and global cerebral blood flow (CBF) were quantified by means of an autoradiographic technique. Compared with the sham condition, both SAH and saline injection resulted in significantly enhanced contraction of cerebral arteries in response to ET-1 and 5-CT. Regional and global CBF were reduced both in the Saline and SAH groups compared with the Sham Group. The mRNA levels for ET(B) and 5-HT(1B) receptors were upregulated after SAH and saline injection compared with the sham procedure. The effects in all parameters were more pronounced for SAH than for saline injection. This study revealed that both the elevation of intracranial pressure and subarachnoid blood per se contribute approximately equally to the late CBF reductions and receptor upregulation following SAH.

Autoradiographic study on the distribution of 241Am in the shell of the freshwater zebra mussel Dreissena polymorpha

Autoradiography was used to identify α-track distributions in a series of shell sections from live mussels Dreissena polymorpha Pallas and dissected shells of dead mussels obtained from laboratory experiments using relatively high concentrations of 241Am in the exposure media, a required condition for successful use of this autoradiographic technique. A comparable distribution of α-tracks was recorded on autoradiographs from both live and dead shells suggesting that metabolism does not lead to any sizable changes in the process of 241Am adsorption (present in the extrapallial fluid) onto the inner surface of shell. Autoradiographs showed a preferential accumulation of 241Am in the organic periostracum, whereas the outer and inner shell layers were characterized by a relatively low α-tracks density. No α-tracks were observed in the central part of the shell in any of the samples. These observations will be useful for the development of a general model to explain bioaccumulation and biosorption processes of radionuclides into mollusk shells.

Wechselwirkungen zwischen pflanzensaugenden Insekten und den von ihnen besogenen Pflanzengeweben

Summary This work was undertaken to find out, using plant physiological methods, the reaction of plant tissue to the penetration by and feeding of plant lice; it showed that spontaneous primary reactions and long term secondary reactions must be distinguished. Shock like disturbances in the transpiration and water intake of the host plant and interference with photosynthesis (by lowering of the assimilation rate) and respiration were established as primary reactions. In the vicinity of the puncture, a raising of the permeability, alteration in viscosity and the raising of the rate of flow of protoplasm could be proved cytophysiologically. The importance of these cytophysiological effects on the feeding of the plant lice is discussed. Besides the already known secondary reactions such as inhibition of growth, gall formation, shoot deformation etc., restriction of flowering and shooting and alterations in osmotic value (alterations in enzyme activity) are described, as well as long term respiration increases, e. g. attack by Quadraspidiotus perniciosus Comst. raises the oxygen consumption to three times its normal value. As it was thought that, besides the mechanical effects, the saliva of plant lice was mainly responsible for phytopathological symptoms, analyses of pure saliva were carried out to determine the enzymes and free amino acids. The free amino acids found had an effect on the respiration, photosynthesis and water balance of the plant tissues and must therefore be largely concerned in the phytopathological effects. The mechanism of saliva secretion was found by the use of a tracer (P32). In phloem feeding aphids (Myzus ascalonicus Donc.), saliva is secreted only with insertion (6–8 min. long) and retraction of the stylets and not in the intervening time no matter how long the insect may feed. In contrast to this, parenchyma feeders secrete more or less continuously and the fundamental differences between the two types of feeding are fully discussed. The mutual effects on plant and aphid, with a parenchyma feeding species, are well illustrated by histochemical and physiological methods, using as an example the bark of Abies pectinata attacked by Adelges piceae. The spread of saliva in the plant is described in detail, using autoradiographic and X-ray techniques. The mechanism of feeding in Myzus ascalonicus is calculated in relation to the relevant pressure conditions and rates of flow.

Translocation and distribution of 32P labelled potassium phosphonate in black pepper ( Piper nigrum L)

The aim of this project was to study the translocation of potassium phosphonate within black pepper ( Piper nigrum L.) and to elucidate the amount of chemical distributed through out the plant based on a tracer technique. Potassium phosphonate is a potential fungicide used against the pathogen Phytophthora capsici, which causes Phytophthora foot rot (Quick wilt) in the plant. Reports indicate that translocation of phosphonate is ambimobile, but there is no visual evidence for this. Potassium phosphonate labelled with radioactive phosphorus, 32P, was applied to the black pepper vine and bush pepper and its translocation to different parts of the pepper plant was studied using an autoradiography technique. The chemical which migrated to different parts of the plant was quantitatively estimated by measuring 32P on a liquid scintillation spectrometer following Cerenkov counting. Only traces of the chemical were lost to the soil through root systems. These results indicate that foliar sprays of potassium phosphonate to pepper could be a viable method of application for soil and foliar pathogens.

Temporal changes in mouse brain fatty acid amide hydrolase activity

Fatty acid amide hydrolase (FAAH) activity is known to mediate the tone of endogenous fatty acid amides including the endocannabinoid anandamide. FAAH is a potential therapeutic target because genetic or pharmacological ablation of FAAH promotes analgesia and anxiolytic effects without disrupting motor coordination. Little is known about the endogenous temporal fluctuations of brain FAAH activity. This is the first comprehensive study examining temporal fluctuations in mouse brain FAAH activity. Regional mouse brain homogenates were generated at the midpoint of the light (“noon”) and dark (“midnight”) cycles. While immunoblots revealed no significant changes ( P>0.05) in regional activity between these two time points, in vitro activity assays detected a subtle 10% reduction ( P<0.05) in cerebellar FAAH activity at midnight. A novel ex vivo autoradiography technique permitted the study of 11 different brain regions, many of which cannot be studied using traditional in vitro methods. The cerebellum and the periaqueductal gray both exhibited significant ( P<0.05) reductions in regional FAAH activity in “midnight” brains. These data confirm the need to account for temporal changes in FAAH activity when therapeutically targeting FAAH.

Comparative chromosome studies in Pinnipedia.

Cells in short-term culture from 12 species of pinnipeds were submitted to chromosome analysis, including, in 11 of them, chromosome measurements and establishment of idiograms. Analysis of variance was carried out on the measurements of 9 phocids. Comparisons between the different karyotypes were made on the basis of conventional chromosome methods as well as autoradiography, G- and C-band techniques. The Pinnipedia are characterized by a pronounced karyotype uniformity. Within the Otarioidea, all otariids so far studied have 2n = 36 and closely agreeing karyotypes, the walrus has 2n = 32. In the Phocidae 2 chromosome numbers occur, 2n = 34 and 2n = 32. The 2n = 32 karyotype has evolved from the primordial 2n = 34 karyotype through fusion of two pairs. Within each of these two groups all karyotypes are virtually identical. The morphological and banding homologies between the otariid and 34 chromosome phocid karyotypes confirm the theory of monophyletic origin of the Pinnipedia.

Application of neutron capture autoradiography to Boron Delivery seeking techniques for selective accumulation of boron compounds to tumor with intra-arterial administration of boron entrapped water-in-oil-in-water emulsion

It is necessary to accumulate the 10B atoms selectively to the tumor cells for effective Boron Neutron Capture Therapy (BNCT). In order to achieve an accurate measurement of 10B accumulations in the biological samples, we employed a technique of neutron capture autoradiography (NCAR) of sliced samples of tumor tissues using CR-39 plastic track detectors. The CR-39 track detectors attached with the biological samples were exposed to thermal neutrons in the thermal column of the JRR3 of Japan Atomic Energy Agency (JAEA). We obtained quantitative NCAR images of the samples for VX-2 tumor in rabbit liver after injection of 10BSH entrapped water-in-oil-in-water (WOW) emulsion by intra-arterial injection via proper hepatic artery. The 10B accumulations and distributions in VX-2 tumor and normal liver of rabbit were investigated by means of alpha-track density measurements. In this study, we showed the selective accumulation of 10B atoms in the VX-2 tumor by intra-arterial injection of 10B entrapped WOW emulsion until 3 days after injection by using digitized NCAR images (i.e. alpha-track mapping).

Measurement of glucose metabolism in rat spinal cord slices with dynamic positron autoradiography

We attempted to measure the regional metabolic rate of glucose (MRglc) in sliced spinal cords in vitro. The thoracic spinal cord of a mature Wister rat was cut into 400-mum slices in oxygenated and cooled (1-4 degrees C) Krebs-Ringer solution. After at least 60 min of preincubation, the spinal cord slices were transferred into double polystyrene chambers and incubated in Krebs-Ringer solution at 36 degrees C, bubbled with 5% O(2)/5% CO(2) gas. To measure MRglc, we used the dynamic positron autoradiography technique (dPAT) with F-18-2-fluoro-2-deoxy-d-glucose ([(18)F]FDG) and the net influx constant of [(18)F]FDG as an index. Uptake curves of [(18)F]FDG were well fitted by straight lines for more than 7 h after the slicing of the spinal cord (linear regression coefficient, r=0.99), indicating a constant uptake of glucose by the spinal cord tissue. The slope (K), which denotes MRglc, is affected by tetrodotoxin, and high K(+) (50 mM) or Ca(2+)-free, high Mg(2+) solution. After 10 min of hypoxia, the K value following reoxygenation was similar to the unloaded control value, but after 45 min of hypoxia, the K value was markedly lower than the unloaded control value, and after >90 min of reoxygenation it was nearly 0. Our results indicate that the living spinal cord slices used retained an activity-dependent metabolism to some extent. This technique may provide a new approach for measuring MRglc in sliced living spinal cord tissue in vitro and for quantifying the dynamic changes in MRglc in response to various interventions such as hypoxia.