Dopaminergic manipulations and its effects on neurogenesis and motor function in a transgenic mouse model of Huntington's disease.

Huntington's disease (HD) is an inherited neurodegenerative disorder that is classically defined by a triad of movement and cognitive and psychiatric abnormalities with a well-established pathology that affects the dopaminergic systems of the brain. This has classically been described in terms of an early loss of dopamine D2 receptors (D2R), although interestingly the treatments most effectively used to treat patients with HD block these same receptors. We therefore sought to examine the dopaminergic system in HD not only in terms of striatal function but also at extrastriatal sites especially the hippocampus, given that transgenic (Tg) mice also exhibit deficits in hippocampal-dependent cognitive tests and a reduction in adult hippocampal neurogenesis. We showed that there was an early reduction of D2R in both the striatum and dentate gyrus (DG) of the hippocampus in the R6/1 transgenic HD mouse ahead of any overt motor signs and before striatal neuronal loss. Despite downregulation of D2Rs in these sites, further reduction of the dopaminergic input to these sites by either medial forebrain bundle lesions or receptor blockade using sulpiride was able to improve both deficits in motor performance and adult hippocampal neurogenesis. In contrast, a reduction in dopaminergic innervation of the neurogenic niches resulted in impaired neurogenesis in healthy WT mice. This study therefore provides evidence that D2R blockade improves hippocampal and striatal deficits in HD mice although the underlying mechanism for this is unclear, and suggests that agents working within this network may have greater effects than previously thought.

Unsaturated macrocyclic dihydroxamic acid siderophores produced by Shewanella putrefaciens using precursor-directed biosynthesis.

To acquire iron essential for growth, the bacterium Shewanella putrefaciens produces the macrocyclic dihydroxamic acid putrebactin (pbH2; [M + H(+)](+), m/zcalc 373.2) as its native siderophore. The assembly of pbH2 requires endogenous 1,4-diaminobutane (DB), which is produced from the ornithine decarboxylase (ODC)-catalyzed decarboxylation of l-ornithine. In this work, levels of endogenous DB were attenuated in S. putrefaciens cultures by augmenting the medium with the ODC inhibitor 1,4-diamino-2-butanone (DBO). The presence in the medium of DBO together with alternative exogenous non-native diamine substrates, (15)N2-1,4-diaminobutane ((15)N2-DB) or 1,4-diamino-2(E)-butene (E-DBE), resulted in the respective biosynthesis of (15)N-labeled pbH2 ((15)N4-pbH2; [M + H(+)](+), m/zcalc 377.2, m/zobs 377.2) or the unsaturated pbH2 variant, named here: E,E-putrebactene (E,E-pbeH2; [M + H(+)](+), m/zcalc 369.2, m/zobs 369.2). In the latter system, remaining endogenous DB resulted in the parallel biosynthesis of the monounsaturated DB-E-DBE hybrid, E-putrebactene (E-pbxH2; [M + H(+)](+), m/zcalc 371.2, m/zobs 371.2). These are the first identified unsaturated macrocyclic dihydroxamic acid siderophores. LC-MS measurements showed 1:1 complexes formed between Fe(III) and pbH2 ([Fe(pb)](+); [M](+), m/zcalc 426.1, m/zobs 426.2), (15)N4-pbH2 ([Fe((15)N4-pb)](+); [M](+), m/zcalc 430.1, m/zobs 430.1), E,E-pbeH2 ([Fe(E,E-pbe)](+); [M](+), m/zcalc 422.1, m/zobs 422.0), or E-pbxH2 ([Fe(E-pbx)](+); [M](+), m/zcalc 424.1, m/zobs 424.2). The order of the gain in siderophore-mediated Fe(III) solubility, as defined by the difference in retention time between the free ligand and the Fe(III)-loaded complex, was pbH2 (ΔtR = 8.77 min) > E-pbxH2 (ΔtR = 6.95 min) > E,E-pbeH2 (ΔtR = 6.16 min), which suggests one possible reason why nature has selected for saturated rather than unsaturated siderophores as Fe(III) solubilization agents. The potential to conduct multiple types of ex situ chemical conversions across the double bond(s) of the unsaturated macrocycles provides a new route to increased molecular diversity in this class of siderophore.

Directing the biosynthesis of putrebactin or desferrioxamine B in Shewanella putrefaciens through the upstream inhibition of ornithine decarboxylase.

To manage iron acquisition in an oxic environment, Shewanella putrefaciens produces the macrocyclic dihydroxamic acid putrebactin (PB) as its native siderophore. In this work, we have established the siderophore profile of S. putrefaciens in cultures augmented with the native PB precursor putrescine and in putrescine-depleted cultures. Compared to base medium, PB increased by two-fold in cultures of S. putrefaciens with 10 mM NaCl and 20 mM exogenous putrescine. In cultures augmented with 1,4-diaminobutan-2-one (DAB), PB decreased with only 0.02-fold PB detectable at 10 mM DAB. As an ornithine decarboxylase (ODC) inhibitor, DAB depleted levels of endogenous putrescine which attenuated downstream PB assembly. Under putrescine-depleted conditions, S. putrefaciens produced as its replacement siderophore the cadaverine-based desferrioxamine B (DFO-B), as characterised by ESI-MS of the Fe(III)-loaded form (m/z(obs) 614.13; m/z(calc) 614.27). A third siderophore, independent of DAB, was observed in low levels. LC/MS Analysis of the Fe(III)-loaded extract gave m/z(obs) 440.93, which, formulated as a 1:1 Fe(III) complex with a macrocyclic dihydroxamic acid, comprising one putrescine- and one cadaverine-based precursor (m/z(calc) 440.14). These results show that the production of native PB or non-native DFO-B by S. putrefaciens can be directed though upstream inhibition of ODC. This approach could be used to increase the molecular diversity of siderophores produced by S. putrefaciens and to map alternative diamine-dependent metabolites.

Ependymal cyst in the cerebrum of an African green monkey (Chlorocebus aethiops).

A focal lesion was detected by magnetic resonance imaging in the right caudal occipital lobe of the cerebrum in an African green monkey (Chlorocebus aethiops). Neurological signs were not observed in this animal. At necropsy examination, an 8mm wedge-shaped intracranial cavity was found, which apparently did not communicate with the ventricles. Microscopically, the inner surface of the cavity was lined by ciliated cuboidal epithelium with positive immunoreactivity for S100 protein, glial fibrillary acidic protein and cytokeratin. Based on the gross, microscopical and immunohistochemical findings the lesion was classified as an ependymal cyst. To the best of our knowledge, this is the first report of an ependymal cyst in an African green monkey.

Studies of iron-uptake mechanisms in two bacterial species of the shewanella genus adapted to middle-range (Shewanella putrefaciens) or antarctic (Shewanella gelidimarina) temperatures.

Iron(III)-uptake mechanisms in bacteria indigenous to the Antarctic, which is the most Fe-deficient continent on Earth, have not been extensively studied. The cold-adapted, Antarctic bacterium, Shewanella gelidimarina, does not produce detectable levels of the siderophore, putrebactin, in the supernatant of Fe(III)-deprived cultures. This is distinct from the putrebactin-producing bacterium from the same genus, Shewanella putrefaciens, which is adapted to middle-range temperatures. The production of putrebactin by S. putrefaciens is optimal, when the pH value of the medium is 7.0. According to the strong positive response from whole cells in the Chrome Azurol S (CAS) agar diffusion assay, Shewanella gelidimarina appears to produce cell-associated siderophores. In the RP-HPLC trace of an Fe(III)-loaded extract from the cell-associated components of S. gelidimarina cultured in media with [Fe(III)] ca. 0 microM, a peak appears at [MeCN] ca. 77%, which decreases in intensity in a parallel experiment in which [Fe(III)] ca. 5 microM, and is barely detectable in Fe(III)-replete media ([Fe(III)] ca. 20 microM). The Fe(III)-dependence of this peak suggests that the attendant species, which is significantly more hydrophobic than putrebactin (RP-HPLC elution: [MeCN] ca. 14%), is associated with Fe(III)-management in S. gelidimarina. This study highlights the diversity in Fe(III)-uptake mechanisms in Shewanella species adapted to different environmental and thermal niches.

Ultra fast symmetry and SIMD-based projection-backprojection (SSP) algorithm for 3-D PET image reconstruction.

Remarkable progress in positron emission tomography (PET) development has occurred in recent years, in hardware, software, and computer implementation of image reconstruction. Recent development in PET scanners such as the high-resolution research tomograph (HRRT) developed by CTI (now Siemens) represents such a case and is capable of greatly enhanced resolution as well as sensitivity. In these PET scanners, the amount of coincidence line data collected contains more than 4.5 x 10(9) coincidence lines of response generated by as many nuclear detectors as 120 000. This formidable amount of data and the reconstruction of this data set pose a real problem in HRRT and have also been of the major bottle neck in further developments of high resolution PET scanners as well as their applications. In these classes of PET scanners, therefore, obtaining one set of reconstructed images often requires many hours of image reconstruction. For example, in HRRT with full data collection in a normal brain scan (using SPAN 3), the image reconstruction time is close to 80 min, making it practically impossible to attempt any list-mode-based dynamic imaging since the image reconstruction time would take many days (as much as 43 h or more for 32-frame dynamic image reconstruction). To remedy this data-handling problem in image reconstruction, we developed a new algorithm based on the symmetry properties of the projection and backprojection processes, especially in the 3-D OSEM algorithm where multiples of projection and back-projection are required. In addition, the single-instruction multiple-data (SIMD) technique also allowed us to successfully incorporate the symmetry properties mentioned above, thereby effectively reducing the total image reconstruction time to a few minutes. We refer to this technique as the symmetry and SIMD-based projection-backprojection (SSP) technique or algorithm and the details of the technique will be discussed and an example of the application of the technique to the HRRT's OSEM algorithm will be presented as a demonstration.

Neural substrates, experimental evidences and functional hypothesis of acupuncture mechanisms.

OBJECTIVES: Although acupuncture therapy has demonstrated itself to be effective in several clinical areas, the underlying mechanisms of acupuncture in general and the analgesic effect in particular are, however, still not clearly delineated. We, therefore, have studied acupuncture analgesic effect through fMRI and proposed a hypothesis, based on the obtained result, which will enlighten the central role of the brain in acupuncture therapy. METHODS: The proposed model, termed as a broad sense hypothalamus-pituitary-adrenal (BS-HPA) axis, was based on our observed neuroimaging results. The model incorporates the stress-induced HPA axis model together with neuro-immune interaction including the cholinergic anti-inflammatory model. RESULTS: The obtained results coupled with accumulating evidence suggest that the central nervous system is essential for the processing of these effects via its modulation of the autonomic nervous system, neuroimmune system and hormonal regulation. CONCLUSIONS: Based on our fMRI study, it appears that understanding the effects of acupuncture within a neuroscience-based framework is vital. Further, we have proposed the broad sense-HPA axis hypothesis which incorporates the experimental results.

The effect of Ting point (tendinomuscular meridians) electroacupuncture on thermal pain: a model for studying the neuronal mechanism of acupuncture analgesia.

OBJECTIVE: The aim of this study was to characterize the role of Ting points (TP) in acute pain management and its potential use in functional imaging studies by quantitatively assessing: (1) the change in peripheral thermal thresholds before and after the electroacupuncture (EA); and (2) the corresponding behavioral feedback of thermal pain stimulation and the de qi sensation of EA. DESIGN: The study design was prospective. SETTINGS/LOCATION: Healthy subjects were recruited for the study at the University of California, San Diego Medical Center. SUBJECTS/INTERVENTIONS: Thirteen (13) healthy subjects were studied. Baseline thermal thresholds (cold and warm sensations and cold and hot pain) were measured at premarked testing sites along the medial aspects of bilateral lower extremities. Five (5) seconds of hot pain (HP) was delivered to the testing sites and the corresponding pain visual analog scale (VAS) scores were recorded. Thirty (30) seconds of EA was delivered via the SP1 and LR1 on the left lower extremities at 5 Hz via a 6-V square-wave stimulator. OUTCOME MEASURES: The VAS scores of the HP and de qi sensation (tingling) during the EA were recorded. The thermal thresholds and VAS scores for the HP and de qi were obtained immediately and both 30 and 60 minutes after the EA. Adaptation testing was also carried out to assess the change in thermal thresholds and the VAS scores of HP without EA. RESULTS: The warm thresholds of bilateral medial calves significantly increased (p < 0.01) after 30 seconds of EA stimulation. The HP VAS score reduced significantly at the ipsilateral calf during EA in comparison to preacupuncture and postacupuncture (p < 0.01) measurements. No significant change in thermal thresholds was noted in the adaptation paradigm. CONCLUSIONS: EA at the TP has an inhibitory effect on the C-fiber afferents. The analgesic benefit observed is most likely A-delta afferent mediated. Further correlation studies in functional imaging may provide defining data for the observed analgesic mechanism.

Total activation change of visual and motor area due to various disturbances.

Brain activation is affected by gradient acoustic noise and various disturbances as well as by other primary tasks. Therefore, we have studied the effects of various disturbances of two different levels of difficulty, that is, weak and strong difficulty levels for primary visual and motor tasks. In the case of visual task with motor and mental disturbances, we found it decreased as motor and mental disturbance difficulty-level increased, compared with the case of without motor and mental disturbances. To the contrary, in the case of motor activity, the total activation of motor cortex with weak and with strong mental disturbance increased as mental disturbance difficulty-levels increased. Therefore, one can conclude that when mental disturbance is added, the visual cortex and motor cortex have an opposite result and when the difficulty-level of the disturbance is increased, the primary tasks are affected more significantly. Although the current observation is preliminary and requires more careful experimental study, it appears that various disturbance effects on brain functions (such as motor and visual cortical responses) produce significant differences in data observations.

New findings of the correlation between acupoints and corresponding brain cortices using functional MRI.

A preliminary study of the correlation between acupuncture points (acupoints) for the treatment of eye disorders suggested by ancient Oriental literature and the corresponding brain localization for vision described by Western medicine was performed by using functional MRI (fMRI). The vision-related acupoint (VA1) is located in the lateral aspect of the foot, and when acupuncture stimulation is performed there, activation of occipital lobes is seen by fMRI. Stimulation of the eye by directly using light results in similar activation in the occipital lobes by fMRI. The experiment was conducted by using conventional checkerboard 8-Hz light-flash stimulation of the eye and observation of the time-course data. This was followed by stimulation of the VA1 by using the same time-course paradigm as visual light stimulation. Results obtained with 12 volunteers yielded very clean data and very close correlations between visual and acupuncture stimulation. We have also stimulated nonacupoints 2 to 5 cm away from the vision-related acupoints on the foot as a control, and activation in the occipital lobes was not observed. The results obtained demonstrate the correlation between activation of specific areas of brain cortices and corresponding acupoint stimulation predicted by ancient acupuncture literature.

A new silent magnetic resonance imaging using a rotating DC gradient.

A new approach to silent MR imaging using a rotating DC gradient has been explored and experimentally studied. Acoustic or sound noise has been one of the problems in examining patients, mainly due to the fast gradient pulsings in interaction with the main magnetic field. The sound noise has been noted to be proportionately louder as the magnetic field strength becomes larger. In this report, we describe a new imaging technique using a mechanically rotating DC gradient coil. The rotating DC gradient coil can effectively replace both phase encoding as well as readout gradient pulsings, and data obtained in this manner can provide a set of projection data that later can be used for projection reconstruction. With some interpolation techniques one can also perform conventional two-dimensional fast Fourier transform image reconstruction. The sound noise intensity compared with the conventional imaging technique, such as the spin-echo sequence, has been reduced down to about -20.7 dB or 117.5 times with this new technique. The experimental pulse sequence and its principle are described and images obtained by the new silent MR imaging technique are reported.

Effects of the acoustic noise of the gradient systems on fMRI: a study on auditory, motor, and visual cortices.

MR acoustic, or sound, noise due to gradient pulsing has been one of the problems in MRI, both in patient scanning as well as in many areas of psychiatric and neuroscience research, such as brain fMRI. Especially in brain fMRI, sound noise is one of the serious noise sources that obscures the small signals obtainable from the subtle changes occurring in oxygenation status in the cortex and blood capillaries. Therefore, we have studied the effects of acoustic, or sound, noise arising in fMR imaging of the auditory, motor, and visual cortices. The results show that the effects of acoustic noise on motor and visual responses are opposite. That is, for motor activity, there is an increased total motor activation, whereas for visual stimulation, the corresponding (visual) cortical activity is diminished substantially when the subject is exposed to a loud acoustic sound. Although the current observations are preliminary and require more experimental confirmation, it seems that the observed acoustic-noise effects on brain functions, such as in the motor and visual cortices, are new observations and could have significant consequences in data observation and interpretation in future fMRI studies.

Analysis of acoustic noise in MRI.

Acoustic or sound noise due to gradient pulsing has been one of the problems in magnetic resonance imaging (MRI), both in patient scanning as well as in many areas of psychiatric and neuroscience research such as functional MRI. Our recent observations in functional MRI for the visual and motor cortex show very different results with sound noise in comparison with the results obtained without sound noise. Although a number of ideas have been suggested in the literature about the possible elimination or reduction of sound noise, progress has been slow due to the basic role of gradient pulsing in MR imaging. Before we tackle the sound-noise-reduction problem, we believe that a systematic study of sound or acoustic noise behavior will provide important information for future endeavors in this area of research in MRI systems, in both commercial and research systems. Therefore, we report on some typical behavior of sound noise observed from MRI scanners and the analyses of their characteristics. Data are obtained both from a commercial MRI scanner (GE Signa 1.5-T EPI system) as well as a research-type MRI scanner (KAIS 2.0-T) developed at a university laboratory setting.

Some new observations on pulse sequence dependent diffusion related edge enhancement in MR microscopy.

Self-diffusion of nuclear spins has been suggested to cause edge enhancement in images especially on a microscopic scale. According to previously published work, theory suggests that edge enhancement is caused by motional narrowing due to the boundaries and spin self-diffusion during the data acquisition period. More careful examination reveals that edge enhancement due to motional narrowing develops only under a few specific conditions. This lack of generality of motional narrowing theory, as well as experimental observations, indicate that edge enhancement due to effects other than motional narrowing alone can exist. It is found that edge enhancement depends greatly on the data acquisition mode; therefore, the images obtained are different depending on the pulse sequence employed. For example, excessive attenuation of DC components due to diffusion can result in edge enhancement in the spin echo signal. However, in the case of FID-like signals, DC components are preserved while positive high frequency parts are attenuated, thereby degrading resolution. The new phenomenon observed has been termed selective spectral suppression since the observed edge enhancement results from the selective attenuation of certain frequency components in the nuclear signals due to diffusion-dependent signal attenuation for a given pulse sequence.

NMR functional imaging using a tailored RF gradient echo sequence: a true susceptibility measurement technique.

The tailored radio frequency gradient echo (TRFGE) technique that has been used in venography (Cho et al., Magn. Reson. Med. 28, 25-38, 1992; Ro, Cho, Magn. Reson. Med. 28, 237, 1992) is applied to functional imaging. The TRFGE technique has the advantage that it is sensitive to the field gradient created by the susceptibility effect, thereby enhancing only the signals from the regions with local field gradient. In addition, the method is insensitive to the in-flow effect, especially from the arterial blood. The latter further simplifies the functional MRI data analysis. The TRFGE sequence is, therefore, suitable for functional MR imaging for which true susceptibility effect measurements are of prime importance. To examine the TRFGE functional imaging systematically, experiments with various imaging parameters such as flip angle alpha, repetition time, and echo time were performed and the results were compared with the data obtained from the conventional gradient echo functional MR imaging. The experimental results shown were all obtained from human volunteers with a 2.0T whole body MRI system.

Applications of NMR imaging to the time-dependent swelling effect in polymers.

The swelling process is often observed in many polymers of interest and is an important phenomenon for the understanding of many biopolymers. This important process, however, is known as poorly understood in the area in polymer science. One of the reasons is that the conventional method of examining the swelling process is often inconsistent. Since it has many important properties including non-invasiveness, Nuclear Magnetic Resonance (NMR) Micro-imaging has become a significant method of analyzing biomaterials as well as biological specimens [1,2]. In the present study, a time-dependent swelling process was observed non-invasively to investigate the polymeric swelling effect using NMR micro-imaging. The present study provides a noble and non-invasive method of measuring the degree of swelling as well as volumetric changes occurring in polymers immersed in liquid. The information obtained relates to both a water ingress process and volumetric changes of polymer specimens. This proposed method also will provide more reliable techniques to ascertain the time-dependent swelling process than the other conventional methods. One of the important aspects of the present study is that the proposed method is a non-invasive technique and is also capable of ascertaining the time-dependent process of the swelling. This may be a new method of measuring the degree of swelling as well as the time-dependent water ingress process in a polymer.

Breakthrough of single-quantum coherence and its elimination in double-quantum filtering.

Breakthrough of single-quantum coherence is shown to occur after application of a double-quantum filter with the conventional four-step phase-cycling scheme. This single-quantum breakthrough is due to the intersequence stimulated echo which has been generated by the radiofrequency pulses in the preceding pulse sequence and appears at the same time as the double-quantum coherence signal in the current pulse sequence. Moreover, the phase of the intersequence stimulated echo is the same as the phase of the double-quantum coherence signal; i.e., the phase of the intersequence stimulated echo is twice the phase change of the radiofrequency pulses in the creation period when their phase is rotated in accordance with the conventional four-step phase-cycling scheme. Consequently, the intersequence stimulated echo passes through the double-quantum filtration in the conventional four-step phase-cycling scheme and gradient pulses. A new phase-cycling scheme which can filter out the single-quantum breakthrough signal is proposed here and its effectiveness is verified experimentally and by computer simulations.

Susceptibility magnetic resonance imaging using spectral decomposition.

Susceptibility differences of materials in magnetic resonance imaging (MRI) usually lead to the intravoxel spin phase variations. Subsequently, the phase variation in the voxel results in a reduction of the signal intensity. This signal intensity reduction is known as the susceptibility effect in MRI and has been studied extensively. In this paper, a new spectral decomposition technique is proposed with which the signal change due to the susceptibility effect can be analyzed. Further, an NMR pulse sequence for the spectral decomposition of the susceptibility was developed and applied to susceptibility imaging of venous blood possessing paramagnetic properties. The computer simulations of the spectral decomposition method and their corresponding experimental results obtained using both a phantom and human volunteers are reported.