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PubMed2003

Impaired sequence learning in carriers of the DYT1 dystonia mutation.

Previous positron emission tomography (PET) studies have shown that nonmanifesting carriers of the DYT1 dystonia mutation express an abnormal pattern of resting glucose metabolism. To determine whether motor behavior is impaired in these subjects, we compared movement and sequence learning in 12 clinically unaffected DYT1 carriers with 12 age-matched controls. Regional differences in brain function during task performance were assessed with simultaneous H(2) (15)O/PET. We found that motor performance was similar in the DYT1 and control groups, with no significant differences in movement time and spatial accuracy measured during each of the tasks. In contrast, sequence learning was reduced in gene carriers relative to controls (p < 0.01). PET imaging during motor execution showed increased activation in gene carriers (p < 0.001, uncorrected) in the left premotor cortex and right supplementary motor area, with concomitant reduction in the posterior medial cerebellum. During sequence learning, activation responses in DYT1 carriers were increased in the left ventral prefrontal cortex, and lateral cerebellum. These findings suggest that abnormalities in motor behavior and brain function exist in clinically nonmanifesting DYT1 carriers. Although localized increases in neural activity may enable normal movement execution in these subjects, this mechanism may not compensate for their defect in sequence learning.

Carrier Proteins↗

PubMed2004

"I have often walked down this street before": fMRI studies on the hippocampus and other structures during mental navigation of an old environment.

The role of the hippocampus in recent spatial memory has been well documented in patients with damage to this structure, but there is now evidence that the hippocampus may not be needed for the storage and recovery of a spatial layout that was experienced long before injury. Such preservation may rely, instead, on a network of dissociable, extra-hippocampal regions implicated in topographical orientation. Using functional magnetic resonance imaging (fMRI), we investigated this hypothesis in healthy individuals with extensive experience navigating in a large-scale urban environment (downtown Toronto). Participants were scanned as they performed mental navigation tasks that emphasized different types of spatial representations. Tasks included proximity judgments, distance judgments, landmark sequencing, and blocked-route problem-solving. The following regions were engaged to varying degrees depending on the processing demands of each task: retrosplenial cortex, believed to be involved in assigning directional significance to locales within a relatively allocentric framework; medial and posterior parietal cortex, concerned with processing space within egocentric coordinates during imagined movement; and regions of prefrontal cortex, present in tasks heavily dependent on working memory. In a second, event-related experiment, a distinct area of inferotemporal cortex was revealed during identification of familiar landmarks relative to unknown buildings in addition to activation of many of those regions identified in the navigation tasks. This result suggests that familiar landmarks are strongly integrated with the spatial context in which they were experienced. Importantly, right medial temporal lobe activity was observed, its magnitude equivalent across all tasks, though the core of the activated region was in the parahippocampal gyrus, barely touching the hippocampus proper.

Adult↗

PubMed1987

[Further development of the diagnostic possibilities of MR tomography. Combination of 3D sequences with a rapid image processing system].

Using 3-D Fourier analysis, it is possible during MR tomography to provide isotropic data of large volumes, ie. uniform spatial resolution with a voxel size of 1 mm3. If one uses a gradient echo sequence with a picture matrix of 256 X 256 X 128 voxels, then examination time is less than 15 minutes. The information contained in this enormous amount of data can be evaluated by means of rapid image analysis. The advantages of this method have been demonstrated by three examples from the head and the spine.

Adenocarcinoma↗

PubMed1994

Spatial control of neuronal cell attachment and differentiation on covalently patterned laminin oligopeptide substrates.

The spatial control of neuronal cell attachment and differentiation via specific receptor mediated interactions, may provide an effective means for the in vitro reconstruction of neuronal cell architecture. In this study, receptor-specific oligopeptide sequences derived from the extracellular matrix (ECM) molecule laminin, a potent neural cell attachment and differentiation promoter were covalently bound on fluorinated ethylene propylene (FEP) films. The degree of receptor-specific cell attachment and the ability to spatially control neurite outgrowth by covalently patterning the oligopeptide sequences on the FEP film surface were assessed. FEP films were first chemically activated with a Radio Frequency Glow Discharge (RFGD) process that covalently replaces the surface fluorine atoms with reactive hydroxyl groups. Oligopeptides containing the YIGSR sequence from the B1 chain of laminin and the water soluble oligopeptide containing the IKVAV sequence (CSRARKQAASIKVAVSADR) from the A chain were covalently bound to the hydroxylated FEP films. Electron Spectroscopy for Chemical Analysis (ESCA) verified the covalent attachment of the oligopeptides to the material surface. The degree of receptor mediated NG108-15 cell attachment on immobilized CDPGYIGSR films was determined using competitive binding media. A 78% reduction in cell attachment was observed on films containing CDPGYIGSR in the cell plating medium. Only a 23% reduction in cell attachment was noted on films plated in medium containing a mock CDPGYIGSK sequence. FEP films immobilized with the IKVAV oligopeptide sequence were shown to mediate PC12 cell attachment and a competitive binding medium also significantly attenuated cell attachment on the immobilized films. The spatial patterning of these oligopeptide sequences to the FEP surface was shown to localize cell attachment and neurite extension on the patterned pathways. The surrounding unmodified FEP surface was inhibitory in serum containing medium and prevented cellular interactions outside the oligopeptide modifications. The spatial immobilization of laminin oligopeptides on FEP films provides a means to organize the attachment and differentiation of neuronal cells in a receptor-specific manner.

Animals↗

PubMed2000

Protection against diverse highly pathogenic H5 avian influenza viruses in chickens immunized with a recombinant fowlpox vaccine containing an H5 avian influenza hemagglutinin gene insert.

A recombinant fowlpox vaccine with an H5 hemagglutinin gene insert protected chickens against clinical signs and death following challenge by nine different highly pathogenic H5 avian influenza viruses. The challenge viruses had 87.3 to 100% deduced hemagglutinin amino acid sequence similarity with the recombinant vaccine, and represented diversely geographic and spatial backgrounds; i.e. isolated from four different continents over a 38 year period. The recombinant vaccine reduced detectable infection rates and shedding titers by some challenge viruses. There was a significant positive correlation in hemagglutinin sequence similarity between challenge viruses and vaccine, and the ability to reduce titers of challenge virus isolated from the oropharynx (r(s)=0.783, P=0.009), but there was no similar correlation for reducing cloacal virus titers (r(s)=-0.100, P=0.78). This recombinant fowlpox-H5 avian influenza hemagglutinin vaccine can provide protection against a variety of different highly pathogenic H5 avian influenza viruses and frequent optimizing of the hemagglutinin insert to overcome genetic drift in the vaccine may not be necessary to provide adequate field protection.

Animals↗

PubMed1999

Cloning, characterisation and expression of the alpha-tubulin genes of the leech, Hirudo medicinalis.

We have isolated two alpha-tubulin cDNAs from the leech, Hirudo medicinalis. Both encode putative proteins of 451 amino-acids which differ from each other at only two positions. Southern blotting suggests that there are only two alpha-tubulin genes in the leech. The genes contain two introns and, because of the extremely high homology of the nucleotide sequence from the second intron to the end of the genes, we have inferred that a gene conversion event about 9.5 million years ago has homogenised the Hirudo alpha-tubulin sequences. Using in situ hybridisation to tissue sections, we have shown that the two genes are probably expressed in all neurons of the leech ganglia and that their spatial distribution remains unchanged during neuronal regeneration. The deduced amino-acid sequences of the leech alpha-tubulins show that they have greatest similarity to those from a platyhelminth, echiuran and mollusc with rather less to arthropod alpha-tubulins. The protein sequences of the leech alpha-tubulins have been compared with representatives of those from across all phyla to determine if any specific feature labels certain isotypes of tubulin for neuronal expression.

Amino Acid Sequence↗

PubMed2004

The performance of young deaf children in spatial and temporal number tasks.

Deaf children tend to fall behind in mathematics at school. This problem may be a direct result of particular experiences in the classroom; for example, deaf children may find it hard to follow teachers' presentations of basic, but nevertheless quite abstract, mathematical ideas. Another possibility is that the problem starts before school: They may either be worse than hearing children at early, nonlinguistic number representations, they may be behind in learning the culturally transmitted number string, or both. This may result in deaf children failing to develop informal problem-solving strategies, which prepare most children for the more formal learning of number and arithmetic that they will have to do at school. We compared 3- and 4-year-old deaf and hearing children's ability to remember and to reproduce the number of items in a set of objects. In one condition, we presented all the items together in a spatial array; in another, we presented them one at a time in a temporal sequence. Deaf children performed as well as the hearing children in the temporal tasks, but outperformed their hearing counterparts in the spatial task. These results suggest that preschool deaf children's number representation is at least as advanced as that of hearing children, and that they are actually better than hearing children at representing the number of objects in spatial arrays. We conclude that deaf children's difficulties with mathematical learning are not a consequence of a delay in number representation. We also conclude that deaf children should benefit from mathematical instruction that emphasizes spatial representation.

Analysis of Variance↗

PubMed2004

[Studies of artifacts with TFE factor increase in heart delayed enhancement MRI sequence IR-T1TFE].

The characteristic of delayed enhancement MRI is high spatial resolution, which makes it possible to evaluate the degree of damage to the myocardium from the inner to the outer membrane of patients with ischemic heart disease. Therefore, this MRI technique is unique in its ability to detect myocardial condition and is necessary to obtain high-quality images. We experienced artifacts induced by TFE factor increase with delayed enhancement of IR-T1TFE. The purpose of this study was to determine the cause of such artifacts. IR-T1TFE changed signal intensity with phase direction as the TFE factor increased. Streak artifacts occurred because signal intensity caused changes with phase direction. Increases in TFE factor prolonged data collection time, such that marked artifacts were created because of changes in signal intensity. Ghost artifacts occurred because signal intensity changed between shots. When the TFE factor was increased, the difference in signal intensity was diminished between shots. The interval of acquired noise decreased in the raw data. Therefore, the interval of ghost artifacts became wider on images.

Artifacts↗

PubMed2005

A hierarchy of associations in hippocampo-cortical systems: cognitive maps and navigation strategies.

In this letter we describe a hippocampo-cortical model of spatial processing and navigation based on a cascade of increasingly complex associative processes that are also relevant for other hippocampal functions such as episodic memory. Associative learning of different types and the related pattern encoding-recognition take place at three successive levels: (1) an object location level, which computes the landmarks from merged multimodal sensory inputs in the parahippocampal cortices; (2) a subject location level, which computes place fields by combination of local views and movement-related information in the entorhinal cortex; and (3) a spatiotemporal level, which computes place transitions from contiguous place fields in the CA3-CA1 region, which form building blocks for learning temporospatial sequences. At the cell population level, superficial entorhinal place cells encode spatial, context-independent maps as landscapes of activity; populations of transition cells in the CA3-CA1 region encode context-dependent maps as sequences of transitions, which form graphs in prefrontal-parietal cortices. The model was tested on a robot moving in a real environment; these tests produced results that could help to interpret biological data. Two different goal-oriented navigation strategies were displayed depending on the type of map used by the system. Thanks to its multilevel, multimodal integration and behavioral implementation, the model suggests functional interpretations for largely unaccounted structural differences between hippocampo-cortical systems. Further, spatiotemporal information, a common denominator shared by several brain structures, could serve as a cognitive processing frame and a functional link, for example, during spatial navigation and episodic memory, as suggested by the applications of the model to other domains, temporal sequence learning and imitation in particular.

Action Potentials↗

PubMed1989

Of mice and men: anatomy of the amelogenin gene.

Mammalian enamel matrix is composed of two principal proteins, the enamelins and amelogenin. Recombinant complementary DNA (cDNA) molecules for the predominant mouse amelogenin have been identified, characterized by direct determination of the DNA sequence, and used as a specific hybridization probe. The spatial- and temporal-restricted pattern for amelogenin gene expression within developing mouse molars has been traced at the level of a single cell using in situ hybridization. The mouse genome has been shown to contain only one copy of the amelogenin (AMEL) gene which is not amplified or rearranged during ameloblast determination. In contrast, the human genome contains two copies of the AMEL gene, one residing on the X chromosome and one upon the Y chromosome. These observations, the availability of specific enamel gene probes coupled with the application of new techniques in molecular biology now afford unique opportunities for the analysis of the molecular basis of inherited defects of human enamel such as amelogenesis imperfecta. Recent advances towards obtaining a physical map and the complete nucleotide sequence for the human genome, as well as the documented developmental biology, defined genetics and transgenic capability of the mouse, suggest that mouse and man are the most relevant and potentially informative models for analysis of normal and abnormal enamel biomineralization.

Amelogenesis Imperfecta↗

PubMed1989

Single amino acid exchanges in the finger domain impair the function of the Drosophila gene Krüppel (Kr).

Mutations in the Drosophila gene Krüppel (Kr) cause deletions of segments in the embryo and also affect Malpighian tubule development. In the hypomorphic Kr alleles that were analyzed, the defects in the segment pattern and the Malpighian tubules are parallel in strength. We have sequenced the DNA of four Kr alleles that show normal spatial and temporal patterns of Kr-encoded protein expression. Three of the four alleles have single base pair mutations that result in a single amino acid change. The exchanges occur in the putative DNA-binding domain of the Kr protein, which is characterized by four repeats of the zinc finger motif. Sequence comparison of the finger motifs helped to define the structural requirements for the folding of the finger domain to some extent. Our data on the Kr mutants support the view that has emerged from the evolutionary analysis.

Alleles↗

PubMed1994

Inaccuracy and instability of sequential movements in Parkinson's disease.

Animal studies suggest that the basal ganglia (BG) provide internal cues to trigger submovements in a movement sequence, with Parkinson's disease (PD) involving a deficiency in this cueing mechanism. However, it is not clear why defective internal cues can produce slow movements, or the extent to which slow movements are indeed the basic movement abnormality or are perhaps a compensatory mechanism for some other primary deficit. In this study we examined a number of the kinematic indices of matched fast movements between PD patients and age-matched controls, performed with and without reductions in visual cues for guidance, in order to delineate the relationship between the internal cue and the kinematic characteristics of these movements. Fourteen patients with PD, and their matched controls, used an electronic pen, which sampled pen-tip position at 200 Hz, and performed a sequence of drawing movements to nine targets upon a WACOM SD 420 graphics tablet. Subjects were trained to perform the movement sequence at a fast speed and were then required to perform the same movement at the same speed with reduced visual cues. Kinematic analysis indicated that, when visual cues were reduced, movements of PD patients became spatially and temporally unstable as they were progressively performed down the sequence. The instability was associated with an abnormal force profile increase in peak movement velocity and target overshoot, which became additive as the submovements progressed. We suggest that defective cue production is the basic deficit in parkinsonian hypokinesia. The defective cue leads to problems synchronising preparatory activity, which then results in abnormalities in movement forces which are characterised by unpredictable and inaccurate movement endpoints. When movements are strung together in a sequence the inaccuracy is additive leading to motor instability.

Aged↗

PubMed1992

Structural requirements for viroid processing by RNase T1.

Viroids are replicated via a rolling circle-like mechanism in which (+) strand oligomeric intermediates have to be cleaved enzymatically to unit-length molecules followed by ligation to mature circles. A transcript of potato spindle tuber viroid, which is still infectious, consists of a monomeric molecule with only 22 additional nucleotides, thus doubling part of the central conserved region of viroids. It was shown that this transcript can be cleaved and ligated in vitro to circles by RNase T1. To elucidate the site and mechanism of processing, 16 different site-specific mutants of this longer-than-unit-length transcript were constructed and analyzed by in vitro processing with RNase T1, infectivity studies, temperature-gradient gel electrophoresis, and structure calculations. The wild-type sequence and several mutated transcripts are able to adopt a particular secondary structure which is the prerequisite for enzymatic cleavage and ligation by RNase T1. This "processing structure" exposes both potential cleavage sites in the nearest spatial neighborhood, thus favoring the subsequent ligation to circles. Those mutated sequences for which the formation of the processing structure is impossible or thermodynamically highly unfavored are not processed. The results demonstrate that the particular structural features of viroids enable them to be cleaved and ligated by one and the same enzyme, RNase T1. The in vitro mechanism may serve as a mechanistic model for cellular processing of viroids.

Base Sequence↗

PubMedDate not supplied

Learning of sensory sequences in cerebellar patients.

A possible role of the cerebellum in detecting and recognizing event sequences has been proposed. The present study sought to determine whether patients with cerebellar lesions are impaired in the acquisition and discrimination of sequences of sensory stimuli of different modalities. A group of 26 cerebellar patients and 26 controls matched for age, sex, handedness, musicality, and level of education were tested. Auditory and visual sensory sequences were presented out of different sensory pattern categories (tones with different acoustic frequencies and durations, visual stimuli with different spatial locations and colors, sequential vision of irregular shapes) and different ranges of inter-cue time intervals (fast and slow). Motor requirements were small, with vocal responses and no time restrictions. Perception of visual and acoustic stimuli was generally preserved in patients and controls. The number of errors was significantly higher in the faster tempo of sequence presentation in learning of sequences of tones of different frequencies and in learning of sequences of visual stimuli of different spatial locations and different colors. No difference in tempo between the groups was shown. The total number of errors between the two groups was identical in the sequence conditions. No major disturbances in acquisition or discrimination of various sensory sequences were observed in the group of cerebellar patients. Sequence learning may be impaired only in tasks with significant motor demands.

Acoustic Stimulation↗

PubMed2003

Use of different proteases working in acidic conditions to improve sequence coverage and resolution in hydrogen/deuterium exchange of large proteins.

The combination of hydrogen exchange and mass spectrometry has been widely used in structural biology, providing views on protein structure and protein dynamics. One of the constraints is to use proteases working at low pH and low temperature to limit back-exchange during proteolysis. Although pepsin works in these conditions and is currently used in such experiments, sequence coverage is not always complete especially for large proteins, and the spatial resolution of the exchange rate is limited by the size of the resulting peptides. In this study we tried two other proteases, protease type XIII from Aspergillus saitoi and protease type XVIII from Rhizhopus species. The penicillin-binding protein X (PBP-2X*), a 77-kDa protein, was selected as a model. Like pepsin, neither of these proteases is really specific, but we found very good reproducibility in the digestion pattern. Compared with using pepsin alone, combining the results of the three independent proteolyses increased the coverage for the peptide mapping, thus avoiding missing some potentially interesting regions of the protein. Furthermore, we obtained a better spatial resolution for deuterium incorporation data, specifying accurately the deuterated regions.

Amino Acid Sequence↗

PubMed2004

Basic assessment of paced activation sequence mapping: implications for practical use.

Some experiences support the use of atrial paced activation sequence mapping, but there is no systematic study assessing its spatial resolution, reproducibility, and influence of pacing parameters. The aim of this study was to evaluate these issues by using a 24-pole catheter positioned at the atrial aspect of the tricuspid and mitral annuli in 15 patients. Bipolar pacing was performed at two sites (right and left atria), 2 cycle lengths (300 and 500 ms) and two outputs (twice and tenfold the late diastolic threshold voltage for 2-ms pulses). The elapsed time between the atrial activation at the two dipoles adjacent to the pacing dipole (activation time [AT]) was measured during each pacing sequence. Changes in cycle length did not modify the AT. The increase in voltage slightly modified the AT (maximum -2 ms at the RA; 95% CI -3 to -1 ms) due to a greater shortening of the conduction time to the dipole located next to the anode. The 95% limits of the intraobserver and interobserver agreements in the AT measurement were -2 to 3 ms and -3 to 3 ms, respectively. The spatial resolution was studied in ten patients by measuring the AT during pacing from each dipole of a 20-pole catheter with a 1-3-1 mm interelectrode distance. The mean AT change was 10 +/- 4 ms per 6 mm of pacing site displacement (95% CI 8-11 ms, range 2.5-20 ms). In conclusion, paced atrial activation sequence analysis is reproducible, accurate, and relatively independent of pacing parameters.

Cardiac Pacing, Artificial↗

PubMed1984

Testing a theory of brain function by computer methods. IV. Comparing spatial and temporal order in the two cerebral hemispheres.

Continuing with a plan for overall testing of the timing and sequencing of brain processes, further testing techniques are introduced for probing a normal adult's short-term central processes, especially those which are employed in reading a short digit span. The tests are administered by means of a personal computer and can provide information on the individual subject's characteristic sequencing and timing of particular central processes. In a subject whose first language is read from left to right, the presented tests reported in this paper can reveal an interesting asymmetry. A temporal display of digits in the right visual field is correctly perceived more readily than in the left. Conversely, a brief, spatial display of digits in the left visual field is correctly reported more often than in the right. It is suggested, therefore, that there are individuals whose left hemisphere is superior in handling a temporal sequence of digits while the right hemisphere is superior in handling a spatial display of digits.

Adult↗

PubMedDate not supplied

Firing characteristics of deep layer neurons in prefrontal cortex in rats performing spatial working memory tasks.

Single cells were recorded with 'tetrodes' in regions of the rat medial prefrontal cortex, including those which are targets of hippocampal afferents, while rats were performing three different behavioral tasks: (i) an eight-arm radial maze, spatial working memory task, (ii) a figure-eight track, delayed spatial alternation task, and (iii) a random food search task in a square chamber. Among 187 recorded units, very few exhibited any evidence of place-specific firing on any of the behavioral tasks, except to the extent that different spatial locations were related to distinct phases of the task. Furthermore, no prefrontal unit showed unambiguous spatially dependent delay activity that might mediate working memory for spatial locations. Rather, the cells exhibited diverse correlates that were generally associated with the behavioral requirements of performing the task. This included firing related to intertrial intervals, onset or end of trials, selection of specific arms on the eight-arm radial maze, delay periods, approach to or departure from goals, and selection of paths on the figure-eight track. Although a small number of cells showed similar behavioral correlates across tasks, the majority of cells showed no consistent correlate when recorded across multiple tasks. Furthermore, some units did not exhibit altered firing patterns in any of the three tasks, while others showed changes in firing that were not consistently related to specific behaviors or task components. These results are in agreement with previous lesion and behavioral studies in rats that suggest a prefrontal cortical role in encoding 'rules' (i.e. structural features) or behavioral sequences within a task but not in encoding allocentric spatial information. Given that the hippocampal projection to this cortical region is capable of undergoing LTP, our data lead to the hypothesis that the role of this projection is not to impose spatial representations upon prefrontal activity, but to provide a mechanism for learning the spatial context in which particular behaviors are appropriate.

Action Potentials↗