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Brain GABA editing by localized in vivo (1)H magnetic resonance spectroscopy.

Editing of GABA by (1)H MRS in a specific brain area is a unique tool for in vivo non-invasive investigation of neurotransmission disorders. Selective GABA detection is achieved using sequences based on double quantum coherence (DQC). Our pulse sequence makes accurate measurements without artefacts due to spatial localization. The sequence was tested on a phantom solution. The effect of vigabatrin, a specific inhibitor of GABA transaminase, was measured in rat brain and GABA detection was performed in vivo in monkey brain using this procedure. Rats were split into two groups. In the control group, the rats had access to water and, in the other group (vigabatrin, VGB, rats), animals were allowed free access to drinking water containing vigabatrin. After 3 weeks of treatment, rats were anesthetized for in vivo NMR spectroscopy investigation. At the end of the experiment, brains were quickly removed, freeze-clamped and extracted with 4% perchloric acid. One part of the acid extract was used for GABA concentrations assessment by ion exchange chromatography with ninhydrin detection. The second was used for high-resolution NMR analysis. By chromatography measurements, the GABA concentration was 1.23+/-0.06 micromol/g for controls, while for vigabatrin-treated rats the GABA concentration was 4.89+/-1.60 micromol/g. The NMR in vivo results were closely correlated with the NMR ex vivo (r=0.99, p<0.01) and chromatography results (r=0.98, p<0.01). The correlation between ex vivo results and chromatography results was also high (r=0.99, p<0.001). This pulse sequence performed GABA editing from a 376 microl voxel located on the right basal ganglia area in a non-human primate brain. This in vivo GABA editing scheme can thus be proposed for accurate measurement of brain GABA concentrations.

Administration, Oral↗

Spatial analysis of volatile organic compounds from a community-based air toxics monitoring network in Deer Park, Texas, USA.

In the summer of 2003, ambient air concentrations of volatile organic compounds (VOCs) were measured at 12 sites within a 3-km radius in Deer Park, Texas near Houston. The purpose of the study was to assess local spatial influence of traffic and other urban sources and was part of a larger investigation of VOC spatial and temporal heterogeneity influences in selected areas of Houston. Seventy 2-h samples were collected using passive organic vapor monitors. Most measurements of 13 VOC species were greater than the method detection limits. Samplers were located at 10 residential sites, a regulatory air monitoring station, and a site located at the centroid of the census tract in which the regulatory station was located. For residential sites, sampler placement locations (e. g., covered porch vs. house eaves) had no effect on concentration with the exception of methyl tertiary-butyl ether (MTBE). Relatively high correlations (Pearson r > 0.8) were found between toluene, ethylbenzene, and o,m,p-xylenes suggesting petroleum-related influence. Chloroform was not correlated with these species or benzene (Pearson r < 0.35) suggesting a different source influence, possibly from process-related activities. As shown in other spatial studies, wind direction relative to source location had an effect on VOC concentrations.

Organic Chemicals↗

Isolation of actin-associated proteins from Caenorhabditis elegans oocytes and their localization in the early embryo.

The actin cytoskeleton plays an important, but poorly understood, role in the development of multicellular organisms. To help illuminate this role, we used actin filament affinity chromatography to isolate actin binding proteins from large quantities of Caenorhabditis elegans oocytes. To examine how these proteins might be involved in early development, we prepared antibodies against some of them and determined their distribution in fixed embryos. Three of these proteins co-localize with different subsets of the embryonic actin cytoskeleton. One co-localizes with actin to all cell cortices. The second oscillates between the nucleus and cortex in a cell-cycle-dependent manner. The third is asymmetrically enriched at the anterior cortex of one-cell embryos, showing a temporal and spatial localization suggestive of a function in generating developmental asymmetry. We conclude that biochemistry is a feasible and useful approach in the study of early C. elegans development, and that the embryonic actin cytoskeleton is regulated in a complex fashion in order to carry out multiple, simultaneous functions.

Actins↗

A genetic oscillator and the regulation of cell cycle progression in Caulobacter crescentus.

Analyses of cell polarity, division, and differentiation in prokaryotes have identified several regulatory proteins that exhibit dramatic changes in expression and spatial localization over the course of a cell cycle. The dynamic behavior of these proteins is often intrinsically linked to their function as polarity determinants.(1-3) In the alpha-proteobacterium, Caulobacter crescentus, the CtrA global transcriptional regulator exhibits a spatially and temporally dynamic expression pattern across the cell cycle. CtrA plays key roles in asymmetric cell division and in the timing of chromosome replication.(3,4) An additional global regulator, GcrA, has recently been discovered that both regulates and is regulated by CtrA.(5) Together, these regulatory proteins create a genetic circuit in which the cellular concentrations of CtrA and GcrA oscillate spatially and temporally to control daughter cell differentiation and cell cycle progression.

Bacterial Proteins↗

Anatomical mapping of white matter hyperintensities (WMH): exploring the relationships between periventricular WMH, deep WMH, and total WMH burden.

BACKGROUND AND PURPOSE: MRI segmentation and mapping techniques were used to assess evidence in support of categorical distinctions between periventricular white matter hyperintensities (PVWMH) and deep WMH (DWMH). Qualitative MRI studies generally identify 2 categories of WMH on the basis of anatomical localization. Separate pathophysiologies and behavioral consequences are often attributed to these 2 classes of WMH. However, evidence to support these empirical distinctions has not been rigorously sought. METHODS: MRI analysis of 55 subjects included quantification of WMH volume, mapping onto a common anatomical image, and spatial localization of each WMH voxel. WMH locations were then divided into PVWMH and DWMH on the basis of distance from the lateral ventricles and correlations, with total WMH volume determined. Periventricular distance histograms of WMH voxels were also calculated. RESULTS: PVWMH and DWMH were highly correlated with total WMH (R2>0.95) and with each other (R2>0.87). Mapping of all WMH revealed smooth expansion from around central cerebrospinal fluid spaces into more distal cerebral white matter with increasing WMH volume. CONCLUSIONS: PVWMH, DWMH, and total WMH are highly correlated with each other. Moreover, spatial analysis failed to identify distinct subpopulations for PVWMH and DWMH. These results suggest that categorical distinctions between PVWMH and DWMH may be arbitrary, and conclusions regarding individual relationships between causal factors or behavior for PVWMH and DWMH may more accurately reflect total WMH volume relationships.

Aged↗

Extreme spatial variability in marine picoplankton and its consequences for interpreting Eulerian time-series.

A high-resolution mesoscale spatial survey of picoplankton in the Celtic Sea, using flow cytometry, reveals cell concentrations of Synechococcus spp. cyanobacteria and heterotrophic bacteria that vary up to 50-fold over distances as short as 12 km. Furthermore, the range of abundances is comparable to that typically found on seasonal scales at a single location. Advection of such spatial variability through a time-series site would therefore constitute a major source of 'error'. Consequently, attempts to model and to investigate the ecology of these globally important organisms in situ must take into account and quantify the hitherto ignored local spatial variability as a matter of necessity.

Animals↗

Affinity regulates spatial range of EGF receptor autocrine ligand binding.

Proper spatial localization of EGFR signaling activated by autocrine ligands represents a critical factor in embryonic development as well as tissue organization and function, and ligand/receptor binding affinity is among the molecular and cellular properties suggested to play a role in governing this localization. We employ a computational model to predict how receptor-binding affinity affects local capture of autocrine ligand vis-a-vis escape to distal regions, and provide experimental test by constructing cell lines expressing EGFR along with either wild-type EGF or a low-affinity mutant, EGF(L47M). The model predicts local capture of a lower affinity autocrine ligand to be less efficient when the ligand production rate is small relative to receptor appearance rate. Our experimental data confirm this prediction, demonstrating that cells can use ligand/receptor binding affinity to regulate ligand spatial distribution when autocrine ligand production is limiting for receptor signaling.

Animals↗

Localized double-quantum filter and correlation spectroscopy experiments.

Problems with in vivo proton spectroscopy include strong water and lipid signals, and the very complicated structures of the spectra. Multiple-quantum experiments can be designed to overcome these difficulties. Using a volume selective refocusing technique, one can demonstrate how multiple-quantum experiments can be performed in a spatially localized manner. Rather than trying to integrate a localization procedure into a particular spectroscopy experiment itself, the approach here is to leave the basic experiment intact and to add a volume selective "block" to the end of the sequence. This "block" must, of course, preserve the integrity of the particular experiment. The volume selective refocusing method, presented here, does just this for a wide variety of proton NMR experiments. The advantage of this approach is that the analysis of the particular experiment is not changed. This is very important considering the complexity involved. Using this procedure, two very elementary multiple-quantum experiments have been performed on a 1.5-T whole-body scanner. These experiments are a localized double-quantum filter experiment (for water suppression) and a localized two-dimensional correlation spectroscopy experiment.

Magnetic Resonance Spectroscopy↗

Epicardial and endocardial localized 31P magnetic resonance spectroscopy: evidence for metabolic heterogeneity during regional ischemia.

Previous studies have noted that myocardial blood flow and high energy phosphates are heterogeneous across the myocardial wall during ischemia. In order to determine whether differences in metabolites between the subendocardium and subepicardium could be detected using 31P magnetic resonance spectroscopy, the Fourier series window (FSW) experiment was implemented on a porcine model of graded regional ischemia. FSW experiments using a planar phantom showed a 46% improvement in localization to the subendocardium compared to a one-pulse experiment. Animal studies of graded ischemia demonstrated a gradient in the phosphocreatine to inorganic phosphate ratio in the myocardium that paralleled the gradient in blood flow. These studies demonstrate the ability of spatially localized 31P magnetic resonance spectroscopy to detect regional changes in myocardial high energy phosphates localized to the subepicardium and subendocardium.

Animals↗

Visuo-manual aiming movements in 6- to 10-year-Old children: evidence for an asymmetric and asynchronous development of information processes.

Sixty children from 6 to 10 years old participated in an open-loop visuo-manual aiming task (Experiment 1). They were asked to point as fast and accurately as possible toward lateralized visual targets. Responses were wrist flexion-extension movements. Results showed non-monotonic changes with age of constant error, reaction time, and movement time. Constant error for targets presented in the right visual field increased between 6 and 8 years and decreased afterward. Reaction time and movement time decreased with age except at 8 years where they tended to increase. The same subjects participated in two control tasks. One task was designed to test the spatial localization of the lateralized visual targets (Experiment 2). Results showed that subjects localized very accurately the targets at all ages. The second control task was designed to test simple reaction time to the same visual stimuli used in the previous tasks (Experiment 3). Results indicate that reaction time decreased linearly with age when no spatial processing is required for the production of the response. The results of the three experiments showed different developmental functions according to the processes involved in each task. Moreover, they suggest that the conversion from visual to motor coordinates undergo a qualitative change at 8 years of age, and that the prevailing process of this conversion is located in the left cerebral hemisphere.

Age Factors↗

Quality control of radiation therapy in multi-institutional randomized clinical trial for localized prostate cancer.

The National Prostatic Cancer Project (NPCP) from 1978 through 1985 compared definitive radiation therapy for Stages B2, C, D1 lesions in those who received only radiation treatment to those who received two years of additional cyclophosphamide (Cytoxan) or estramustine phosphate (Emcyt) chemotherapy. Two hundred fifty-four patients were entered and 229 evaluated for compliance of the spatial localization of the prostate through review of the simulation and port films. In 78 per cent this was satisfactory, whereas in 12 per cent it was unsatisfactory, and another 10 per cent were not evaluable. The principle cause of an unsatisfactory rating was failure to adequately cover the prostatic target volume, especially the apex which was found to be variable in location. Routine use of retrograde urethrocystography is urged as part of the localization method in patients to receive definitive external beam radiation therapy for prostate cancer. The role and impact of quality assurance programs for radiotherapy in cooperative clinical study groups is reviewed and discussed.

Clinical Trials as Topic↗

Electrophysiological evidence for an early(pre-attentive) information processing deficit in patients with right hemisphere damage and unilateral neglect.

Patients with right hemisphere damage and contralesional neglect are often unaware of visual, auditory or tactile stimuli occurring on their left side. In an effort to understand the contribution of pre-attentive processes to this phenomenon, we examined the processing of the pitch, duration and spatial location of auditory stimuli using an electrophysiological probe, the mismatch negativity (MMN). This event-related brain potential indexes the integrity of cerebral processes that respond automatically to deviations from regularity in the acoustic environment. We compared the MMN elicited by right- and left-sided deviant stimuli in 10 patients with left unilateral neglect and 10 age-matched healthy volunteers, exploring an anticipated dissociation between the processing of spatial localization of sounds and the processing of the other auditory dimensions. Across dimensions, the MMN elicited by deviance occurring to the left of the patients was reduced relative to that elicited by deviance occurring to the right. This effect was robust for spatial location, and less so for pitch, whereas the processing of stimulus duration was not significantly affected by the side of stimulation. In healthy subjects, deviance in either side elicited similar MMN. We suggest that an early deficit in detecting changes in the environment hampers the involuntary triggering of attention in those patients and discuss the specific role of encoding spatial location in the establishment of conscious awareness.

Adult↗

Local sequence-structure motifs in RNA.

Ribonuclic acid (RNA) enjoys increasing interest in molecular biology; despite this interest fundamental algorithms are lacking, e.g. for identifying local motifs. As proteins, RNA molecules have a distinctive structure. Therefore, in addition to sequence information, structure plays an important part in assessing the similarity of RNAs. Furthermore, common sequence-structure features in two or several RNA molecules are often only spatially local, where possibly large parts of the molecules are dissimilar. Consequently, we address the problem of comparing RNA molecules by computing an optimal local alignment with respect to sequence and structure information. While local alignment is superior to global alignment for identifying local similarities, no general local sequence-structure alignment algorithms are currently known. We suggest a new general definition of locality for sequence-structure alignments that is biologically motivated and efficiently tractable. To show the former, we discuss locality of RNA and prove that the defined locality means connectivity by atomic and non-atomic bonds. To show the latter, we present an efficient algorithm for the newly defined pairwise local sequence-structure alignment (lssa) problem for RNA. For molecules of lengthes n and m, the algorithm has worst-case time complexity of O(n2 x m2 x max(n,m)) and a space complexity of only O(n x m). An implementation of our algorithm is available at http://www.bio.inf.uni-jena.de. Its runtime is competitive with global sequence-structure alignment.

Algorithms↗

[Functional asymmetry of the neocortex electrical activity during food conditioning in dogs].

Four dogs were trained to perform a conditioned alimentary response to a sound stimulus. The EEG was recorded from six pairs of chronically implanted neocortical electrodes. The EEG spectra and coherence functions between the neighboring derivations of each of the hemispheres were analyzed in the theta, alpha, beta 1 and beta 2 frequency ranges. At the first stages of conditioning, the percent of cases increased when the highest mean values of EEG frequency were localized in the left hemisphere. Later on the percent of cases, when the mean coherence values in the left hemisphere were higher than in the right hemisphere, also increased. At the stage of conditioned response stabilization, this asymmetry either disappeared or the right hemisphere became more active than the left one. The spatial localization of the maximal values of the EEG frequency was different for different frequency ranges. The highest values in the beta 1 range were more frequently registered in the posterior cortical regions and in the beta 2 range they were revealed, predominantly, in the anterior areas. The maximal values of coherence dominated in the anterior regions and their spatial distribution was similar for different frequencies. Thus, the initial stages of conditioning are accompanied by activation of the left hemisphere.

Animals↗

Two-dimensional imaging with a single-sided NMR probe.

A new low field unilateral NMR sensor equipped with a two-dimensional gradient coil system was built. A new NMR-MOUSE concept using a simple bar magnet instead of the classical U-shaped geometry was used to produce magnetic field profiles comparatively homogeneous in extended lateral planes defining a suitable field of view for 2D spatial localization. Slice selection along the depth direction is obtained by means of the highly constant static magnetic field gradient produced by this magnet geometry. Implementing a two-dimensional phase-encoding imaging method 2D cross sections of objects were obtained with high spatial resolution. By retuning the probe it was possible to change the depth of the selected slice obtaining a 3D imaging method. The details of the construction of the new device are presented together with imaging tests to show the quality of space encoding.

Equipment Design↗

Epicardial but not endocardial premature stimulation initiates ventricular tachyarrhythmia in canine in vitro model of long QT syndrome.

OBJECTIVES: To explore the mechanism, we tested the hypothesis that premature epicardial stimulation transiently increased the dispersion of repolarization leading to VT. BACKGROUND: Premature stimulation initiated ventricular tachycardia (VT) when applied to the epicardium but not to the endocardium in a canine model of long QT syndrome (LOTS). METHODS: We optically mapped action potentials (APs) on the cut-exposed transmural surfaces of isolated wedges of canine ventricular walls perfused with anemone toxin II (ATX-II), which produced type 3 LQTS with an asymmetrical transmural profile of repolarization that was earliest in the epicardium and latest in deep subendocardium. RESULTS: Earliest excitable epicardial stimulation triggered VT in 5 of 18 wedges receiving > or =5 nmol/L ATX-II by direct activation of epicardium, which delayed repolarization in the still refractory midmyocardium and further enhanced the dispersion of repolarization. These VTs were initiated 197 +/- 72 ms (n = 10) after the premature stimulation, from focal regions of earliest repolarization downstream to the steepest local spatial gradients of repolarization, and maintained by new focal activation and reentry. Transmural differences in the cycle lengths of activations altered conduction pathways and resulted in torsades de pointes-like polymorphic VT. In contrast, VTs were not initiated by endocardial stimulation at the same premature intervals or when ATX-II was < or =2.5 nmol/L. Failed VT initiation was associated with significantly lower maximum local gradient of repolarization. CONCLUSIONS: Heterogeneic repolarization in LQTS provides a transmural asymmetrical substrate for the earliest excitable epicardial, but not endocardial, stimulation to further delay midmyocardial repolarization and produce a steep spatial gradient of repolarization potential initiating torsades de pointes-like polymorphic VT.

Action Potentials↗

Melatonin receptor mRNA localization and rhythmicity in the retina of the domestic chick, Gallus domesticus.

The indoleamine hormone melatonin is synthesized and released by photoreceptors during the night within the chick retina, and confers timing information to modulate retinal physiology. Three subtypes of melatonin receptor with nearly identical pharmacological profiles have been described in chickens and are present in the retina. In this study, the spatial localization and temporal pattern of the mRNA for each of these receptors within the retina are described. The localization and rhythmicity of receptor mRNA were analyzed using in situ hybridization and RNase protection assay, respectively, with probes against specific nucleotide sequences encoding these receptors. Mel1A and Mel1C receptor mRNA have similar patterns of expression, primarily in the inner segments of photoreceptors, vitread portion of the inner nuclear layer, and in the retinal ganglion cell layer. Mel1B receptor mRNA is expressed at higher levels in the retina, with expression in photoreceptors, throughout the inner nuclear layer, and in the ganglion cell layer. Mel1A receptor mRNA is rhythmic in both light:dark (LD) cycles and in constant darkness (DD); Mel1A peaks during midday and mid-subjective day, respectively. Mel1C receptor mRNA is also rhythmically expressed in LD, but with a lower amplitude, such that transcript is high during the day and low during the night. In DD, Mel1C rhythms become 180 deg out of phase with a slight increase at night. Mel1B mRNA expression was highly variable and arrhythmic.

Animals↗

From local to global spatiotemporal chaos in a cardiac tissue model.

Two kinds of chaos can occur in cardiac tissue, chaotic meander of a single intact spiral wave and chaotic spiral wave breakup. We studied these behaviors in a model of two-dimensional cardiac tissue based on the Luo-Rudy I action potential model. In the chaotic meander regime, chaos is spatially localized to the core of the spiral wave. When persistent spiral wave breakup occurs, there is a transition from local to global spatiotemporal chaos.

Action Potentials↗