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Mapping the naked neck (NA) and polydactyly (PO) mutants of the chicken with microsatellite molecular markers.

The bulked segregant analysis methodology has been used to map, with microsatellite markers, two morphological mutations in the chicken: polydactyly (PO) and naked neck (NA). These autosomal mutations show partial dominance for NA, and dominance with incomplete penetrance for PO. They were mapped previously to different linkage groups of the classical map, PO to the linkage group IV and NA being linked to the erythrocyte antigen CPPP. An informative family of 70 offspring was produced by mating a sire, heterozygous for each of the mutations, to 7 dams homozygous recessive for each locus. Three DNA pools were prepared, pool PO included 20 chicks exhibiting at least one extra-toe, pool NA included 20 non-polydactyly chicks showing the typical phenotype associated with heterozygosity for the naked neck mutation, and pool NP included 20 chicks exhibiting neither of the mutant phenotypes. Typings were done on an ABI-373 automatic sequencer with 147 microsatellite markers covering most of the genome. An unbalanced distribution of sire marker alleles were detected between pool PO, and pools NA and NP, for two markers of chromosome 2p, MCW0082 and MCW0247. A linkage analysis taking into account the incomplete penetrance of polydactyly (80% ) was performed with additional markers of this region and showed that the closest marker to the PO locus was MCW0071 (5 cM, lod score = 9). MCW0071 lies within the engrailed gene EN2 in the chicken. In the mouse, the homologous gene maps on chromosome 5, close to the hemimelic extra-toes mutation Hx. In the case of the NA locus, markers of chromosome 3 were selected because CPPP was mapped on this chromosome. Analysis of individual typings showed a linkage of 5.7 cM (lod score = 13) between the NA locus and ADL0237 in the distal region of chromosome 3q. These results contribute to connecting the former classical map to the molecular genetic map of the chicken, and open the way to the identification of the molecular nature of two developmental mutations of the chicken that are known to occur in many breeds of chickens.

Journal Article↗

Mapping corpus callosum deficits in autism: an index of aberrant cortical connectivity.

BACKGROUND: Volumetric studies have reported reductions in the size of the corpus callosum (CC) in autism, but the callosal regions contributing to this deficit have differed among studies. In this study, a computational method was used to detect and map the spatial pattern of CC abnormalities in male patients with autism. METHODS: Twenty-four boys with autism (aged 10.0 +/- 3.3 years) and 26 control boys (aged 11.0 +/- 2.5 years) underwent a magnetic resonance imaging (MRI) scan at 3 Tesla. Total and regional areas of the CC were determined using traditional morphometric methods. Three-dimensional (3D) surface models of the CC were also created from the MRI scans. Statistical maps were created to visualize morphologic variability of the CC and to localize regions of callosal thinning in autism. RESULTS: Traditional morphometric methods detected a significant reduction in the total callosal area and in the anterior third of the CC in patients with autism; however, 3D maps revealed significant reductions in both the splenium and genu of the CC in patients. CONCLUSIONS: Statistical maps of the CC revealed callosal deficits in autism with greater precision than traditional morphometric methods. These abnormalities suggest aberrant connections between cortical regions, which is consistent with the hypothesis of abnormal cortical connectivity in autism.

Adolescent↗

Effect of focal and nonfocal cerebral lesions on functional connectivity studied with MR imaging.

BACKGROUND AND PURPOSE: Functional connectivity MR (fcMR) imaging is used to map regions of brain with synchronous, regional, slow fluctuations in cerebral blood flow. We tested the hypothesis that focal cerebral lesions do not eradicate expected functional connectivity. METHODS: Functional MR (fMR) and fcMR maps were acquired for 12 patients with focal cerebral tumors, cysts, arteriovenous malformations, or in one case, agenesis of the corpus callosum. Task activation secondary to text listening, finger tapping, and word generation was mapped by use of fMR imaging. Functional connectivity was measured by selecting "seed" voxels in brain regions showing activation (based on the fMR data) and cross correlating with every other voxel (based on data acquired while the subject performed no task). Concurrence of the fMR and fcMR maps was measured by comparing the location and number of voxels selected by both methods. RESULTS: Technically adequate fMR and fcMR maps were obtained for all patients. In patients with focal lesions, the fMR and fcMR maps correlated closely. The fcMR map generated for the patient with agenesis of the corpus callosum failed to reveal functional connectivity between blood flow in the left and right sensorimotor cortices and in the frontal lobe language regions. Nonetheless, synchrony between blood flow in the auditory cortices was preserved. On average, there was 40% concurrence between all fMR and fcMR maps. CONCLUSION: Patterns of functional connectivity remain intact in patients with focal cerebral lesions. Disruption of major neuronal networks, such as agenesis of the corpus callosum, may diminish the normal functional connectivity patterns. Therefore, functional connectivity in such patients cannot be fully demonstrated with fcMR imaging.

Agenesis of Corpus Callosum↗

Low-frequency signal changes reflect differences in functional connectivity between good readers and dyslexics during continuous phoneme mapping.

The current fMRI study investigated correlations of low-frequency signal changes in the left inferior frontal gyrus, right inferior frontal gyrus and cerebellum in 13 adult dyslexic and 10 normal readers to examine functional networks associated with these regions. The extent of these networks to regions associated with phonological processing (frontal gyrus, occipital gyrus, angular gyrus, inferior temporal gyrus, fusiform gyrus, supramarginal gyrus and cerebellum) was compared between good and dyslexic readers. Analysis of correlations in low-frequency range showed that regions known to activate during an "on-off" phoneme-mapping task exhibit synchronous signal changes when the task is administered continuously (without any "off" periods). Results showed that three functional networks, which were defined on the basis of documented structural deficits in dyslexics and included regions associated with phonological processing, differed significantly in spatial extent between good readers and dyslexics. The methodological, theoretical and clinical significance of the findings for advancing fMRI research and knowledge of dyslexia are discussed.

Adult↗

Mapping of the amino acids in the cytoplasmic loop connecting helices C and D in rhodopsin. Chemical reactivity in the dark state following single cysteine replacements.

The cytoplasmic loop connecting helices C and D in rhodopsin is a part of the region involved in protein-protein interactions during signal transduction. To probe the structure of the CD loop, we have replaced, one at a time, the amino acids 136-150 by cysteine residues. The cysteine substitution mutants contained only the introduced single reactive cysteines and were prepared from a base opsin mutant that retained only the three intradiscal cysteines. All of the cysteine substitution mutants formed the characteristic rhodopsin chromophore (lambda max, 500 nm) with 11-cis-retinal. They showed normal photobleaching characteristics and activated transducin in a light-dependent manner, albeit at lower levels than the wild-type pigment. The newly introduced cysteines in the substitution mutants all underwent alkylation in the dark with the membrane-permeant sulfhydryl reagent N-ethylmaleimide, but with varying rates. The cysteine substitution mutants also showed prominent differences in alkylation with membrane-impermeant N-polymethylenecarboxylmaleimides of various alkyl chain lengths. Notably, derivatization of the cysteines in the mutants was not observed with the polar sulfhydryl reagents iodoacetic acid or iodoacetamide. These findings highlight intrinsic differences in both the reactivity and accessibility of the different cysteine residues in the CD loop and support the important role for a structure in the second cytoplasmic region of rhodopsin.

Amino Acid Sequence↗

Topology representing network enables highly accurate classification of protein images taken by cryo electron-microscope without masking.

In single-particle analysis, a three-dimensional (3-D) structure of a protein is constructed using electron microscopy (EM). As these images are very noisy in general, the primary process of this 3-D reconstruction is the classification of images according to their Euler angles, the images in each classified group then being averaged to reduce the noise level. In our newly developed strategy of classification, we introduce a topology representing network (TRN) method. It is a modified method of a growing neural gas network (GNG). In this system, a network structure is automatically determined in response to the images input through a growing process. After learning without a masking procedure, the GNG creates clear averages of the inputs as unit coordinates in multi-dimensional space, which are then utilized for classification. In the process, connections are automatically created between highly related units and their positions are shifted where the inputs are distributed in multi-dimensional space. Consequently, several separated groups of connected units are formed. Although the interrelationship of units in this space are not easily understood, we succeeded in solving this problem by converting the unit positions into two-dimensional (2-D) space, and by further optimizing the unit positions with the simulated annealing (SA) method. In the optimized 2-D map, visualization of the connections of units provided rich information about clustering. As demonstrated here, this method is clearly superior to both the multi-variate statistical analysis (MSA) and the self-organizing map (SOM) as a classification method and provides a first reliable classification method which can be used without masking for very noisy images.

Algorithms↗

Use of novel nonfluoroscopic three-dimensional electroanatomic mapping system to monitor and analyze heart surgery in animal models.

BACKGROUND: The new method of three-dimensional (3D) electroanatomic mapping was presented as an important tool for cardiac imaging and intervention. We present herein the first use of this technology for the monitoring, analysis, and development of cardiac surgery at the preclinical stage. METHODS: The method is based on utilizing a locatable catheter connected to an endocardial mapping and navigating system, to accurately establish the location and orientation of the tip of the mapping catheter and simultaneously record its local electrogram. The 3D geometry of the beating cardiac chamber is reconstructed in real time. The system was tested on six goats that underwent dynamic cardiomyoplasty. Two maps of each animal were performed: preoperative and postoperative during the stimulation protocol of the skeletal muscle. RESULTS: The electroanatomic mapping system provided detailed maps of the left ventricle during the stimulation protocol, which demonstrated a striking geometric difference between the assisted and the unassisted beats. These geometric changes are best described by referring to left ventricular long-axis movements (22.3 +/- 3.8 degrees vs 3.4 +/- 1.6 degrees, p < 0.001), center-of-mass movements (10.4 +/- 3.0 mm vs 3.9 +/- 1.6 mm, p < 0.005), and the changes in upward movement viewed along the base (7.9 +/- 1.9 mm vs 3.6 +/- 1.7 mm, p < 0.01), middle (13.8 +/- 4.0 mm vs 7.3 +/- 1.8 mm, p < 0.005), and the apex of the heart (28.1 +/- 4.5 vs 5.3 +/- 2.3 mm, p < 0.001) [mean +/- SD]. CONCLUSIONS: The 3D electroanatomic mapping system allows detailed reconstruction of the left ventricular geometry and a clear view of the difference between the assisted and the unassisted beats. This novel monitoring system may serve as an important tool for the analysis and development of new techniques in cardiac surgery.

Animals↗

Identification of candidate sequences that determine virulence in Coxsackievirus B4.

We have previously shown that a major determinant of virulence for coxsackievirus B4 mapped to the 5' end of the viral genome. Comparison of the corresponding cDNA sequences of a virulent and a non-virulent virus has allowed the identification of candidate determinants of virulence in the 5' untranslated region and the capsid proteins VP1, VP2 and VP4. Thirteen nucleotide substitutions were observed in a region spanning 3298 nucleotides. Four mutations were detected in the non-coding region. Of the remaining nine mutations, four were silent while five resulted in amino acid substitutions in VP1, VP2 and VP4. The amino acid substitutions in the virulent virus were analyzed in relation to the three-dimensional structures of the capsid proteins of poliovirus. Two substitutions mapped to the amino termini of VP1 and VP4. Of the two substitutions observed in VP2, one mapped to the large loop that connects beta strand E with the radial helix on the back surface of the eight-stranded antiparallel beta barrel while the other mapped to beta strand G. One amino acid substitution in VP1 mapped to the loop connecting beta strands D and E at a site close to a major determinant of attenuation in poliovirus type 2.

Amino Acid Sequence↗

Application of Brodmann's area templates for ROI selection in white matter tractography studies.

Brodmann's areas are part of the common vernacular used by neuroscientists to indicate specific location of brain activity in functional brain imaging studies. Here, we have employed a template based on the Brodmann's areas as a means of compartmentalizing underlying white matter pathways. White matter tractography was performed on the diffusion tensor data of sixteen subjects using a streamline tracking technique with Runge-Kutta integration. After co-registration, the Brodmann template was utilized for ROI selection. Tracts were segmented based on their termination in a particular area of the template. Binary masks were generated based on the tractography segmentation for a given Brodmann's area in each individual subject. Following registration to a normalized coordinate space, the binary masks were averaged, generating a map that estimates the probability of tractography connectivity for particular white matter pathways to a specific Brodmann's area. The probability maps were color-coded and overlaid on anatomical images to provide perspective. In this study, particular attention was given to the areas of the frontal cortex. A composite map of these areas was generated by assigning each voxel to the Brodmann's area with the highest probability of connectivity, based on the average results. The average maps generated with this method reveal consistent patterns of connectivity across subjects. The use of a normalized template for ROI selection automates the process of segmenting tractography data, making it particularly useful for multi-subject studies. In the future, this method could be used to help elucidate relationships between function and anatomical structure.

Adult↗

Synaptic connectivity in hippocampal neuronal networks cultured on micropatterned surfaces.

Embryonic rat hippocampal neurons were grown on patterned silane surface in order to organize synapse formations in a controlled manner. The surface patterns were composed of trimethoxysilylpropyl-diethylenetriamine (DETA) lines separated by tridecafluoro-1,1,2,2-tetrahydrooctyl-1-dimethylchlorosilane (13F) spaces. Pre- and post-synaptic specializations were identified by immunostaining for synapsin I and microtubule-associated protein-2 (MAP-2). Functional synaptic connections were examined by recording simultaneously from pairs of neurons using the whole-cell configuration of the patch-clamp technique. Spontaneous and evoked synaptic currents were recorded in neurons cultured for 2-14 days. The formation of functional connections was accompanied by the appearance of spontaneous synaptic currents (SSCs), which could be detected after approximately 3 days in culture in the absence of evoked synaptic currents (ESCs). ESCs were detected only after approximately 7 days in culture, mostly in the form of unidirectional synaptic connections. Other forms of synaptic connectivity, such as bidirectional and autaptic connections, were also identified. Both transient GABAergic and glutamatergic signals mediated the transmissions between communicating cells. These results demonstrate the combination of various types of synaptic connections forming simple and complex networks in neurons cultured on line (DETA)-space (13F) patterns. Finally, precisely synchronized SSCs were recorded in neuron pairs cultured on pattern indicating the existence of a fast-acting feedback mechanism mediated by pre-synaptic GABA(A) receptors.

Alkanes↗

Cognitive interference is associated with neuronal marker N-acetyl aspartate in the anterior cingulate cortex: an in vivo (1)H-MRS study of the Stroop Color-Word task.

The neurobiology of cognitive interference is unknown. Previous brain imaging studies using the Stroop Color-Word (SCW) task indicate involvement of the cingulate cortex cognitive division. The present study examines interrelationships between regional brain N-Acetyl aspartate (NAA) levels (as identified by in vivo proton magnetic resonance spectroscopy in the right and left anterior cingulate cortex (ACC), dorsolateral prefrontal cortex, orbitofrontal cortex and thalamus) and cognitive interference (as measured by the SCW task) in 15 normal subjects. The results show that brain chemistry depends on cognitive interference levels (high vs low). Reduction of NAA levels was demonstrated in the right ACC (ie, cognitive midsupracallosal division) of high interference subjects, as compared to the low interference group (P < 0.01, two-tailed t-test). Chemical-cognitive relationships were analyzed by calculating correlations between regional NAA levels and the SCW task scores. Cognitive interference was highly correlated with the right anterior cingulate NAA (r = 0.76, P < 0.001), and was unrelated to other studied regional NAA, including the left ACC (P < 0.025; comparing the difference between r values in the right and left ACC). The interrelationships between NAA across brain regions were examined using correlation analysis (square matrix correlation maps), which detected different connectivity patterns between the two groups. These findings provide evidence of ACC involvement in cognitive interference suggesting a possibility of neuronal reorganization in the physiological mechanism of interference (most likely due to genetically predetermined control of the number of neurons, dendrites and receptors, and their function). We conclude that spectroscopic brain mapping of NAA, the marker of neuronal density and function, to the SCW task measures differentiates between high and low interference in normal subjects. This neuroimaging/cognitive tool may be useful for documentation of interference in studying cognitive control mechanisms, and in diagnosis of neuropsychiatric disorders where dysfunction of cingulate cortex is expected.

Adult↗

Molecular mapping of periodontal tissues using infrared microspectroscopy.

BACKGROUND: Chronic periodontitis is an inflammatory disease of the supporting structures of the teeth. Infrared microspectroscopy has the potential to simultaneously monitor multiple disease markers, including cellular infiltration and collagen catabolism, and hence differentiate diseased and healthy tissues. Therefore, our aim was to establish an infrared microspectroscopy methodology with which to analyze and interpret molecular maps defining pathogenic processes in periodontal tissues. METHODS: Specific key cellular and connective tissue components were identified by infrared microspectroscopy and using a chemical imaging method. RESULTS: Higher densities of DNA, total protein and lipid were revealed in epithelial tissue, compared to the lower percentage of these components in connective tissue. Collagen-specific tissue mapping by infrared microspectroscopy revealed much higher levels of collagen deposition in the connective tissues compared to that in the epithelium, as would be expected. Thus inflammatory events such as cellular infiltration and collagen deposition and catabolism can be identified by infrared microspectroscopy. CONCLUSION: These results suggest that infrared microspectroscopy may represent a simple, reagent-free, multi-dimensional tool with which to examine periodontal disease etiology using entirely unprocessed tissue sections.

Journal Article↗

Molecular similarity indices in a comparative analysis (CoMSIA) of drug molecules to correlate and predict their biological activity.

An alternative approach is reported to compute property fields based on similarity indices of drug molecules that have been brought into a common alignment. The fields of different physicochemical properties use a Gaussian-type distance dependence, and no singularities occur at the atomic positions. Accordingly, no arbitrary definitions of cutoff limits and deficiencies due to different slopes of the fields are encountered. The fields are evaluated by a PLS analysis similar to the CoMFA formalism. Two data sets of steroids binding to the corticosteroid-binding-globulin and thermolysin inhibitors were analyzed in terms of the conventional CoMFA method (Lennard-Jones and Coulomb potential fields) and the new comparative molecular similarity indices analysis (CoMSIA). Models of comparative statistical significance were obtained. Field contribution maps were produced for the different models. Due to cutoff settings in the CoMFA fields and the steepness of the potentials close to the molecular surface, the CoMFA maps are often rather fragmentary and not contiguously connected. This makes their interpretation difficult. The maps obtained by the new CoMSIA approach are superior and easier to interpret. Whereas the CoMFA maps denote regions apart from the molecules where interactions with a putative environment are to be expected, the CoMSIA maps highlight those regions within the area occupied by the ligand skeletons that require a particular physicochemical property important for activity. This is a more significant guide to trace the features that really matter especially with respect to the design of novel compounds.

Models, Molecular↗

Three-dimensional electromechanical mapping: imaging in the operating room of the future.

BACKGROUND: Three-dimensional electromechanical mapping has previously been shown to be a clinically important tool for cardiac imaging and intervention. We hypothesized that this technology may be beneficial as an intraoperative modality for assessing cardiac hemodynamics and viability during cardiac surgery. We report here the use of this technology as an imaging modality for intraoperative cardiac surgery. METHODS: The tip of a locatable catheter connected to an endocardial mapping and navigating system is accurately localized while simultaneously recording local electrical and mechanical functions. Thus the three-dimensional geometry of the beating cardiac chamber is reconstructed in real time. The system was tested on 6 goats that underwent acute dynamic cardiomyoplasty and on 5 dogs that underwent left anterior descending (LAD) coronary artery ligation. RESULTS: The electromechanical mapping system provided an accurate three-dimensional reconstruction of the beating left ventricle during cardiomyoplasty. After the wrapping procedure, significant end-diastolic area reduction was noted in the base and mid parts of the heart (948 +/- 194 mm2 vs 1245 +/- 33 mm2, p = 0.021; and 779 +/- 200 mm2 vs 1011 +/- 80 mm2, p = 0.016). The area of the cross-section of the apex did not change during the operation. Acute infarcted tissue was characterized 3 days after LAD ligation by concomitant deterioration in both electrical and mechanical function. CONCLUSIONS: By providing both a clear view of the anatomical changes that occur during cardiac surgery, and an accurate assessment of tissue viability, electroanatomic mapping may serve as an important adjunct tool for imaging and analysis of the heart during cardiac surgery

Animals↗

N-methyl-D-aspartate receptor antagonists disrupt the formation of a mammalian neural map.

The topographic ordering of retinal connections in the rat superior colliculus emerges during early postnatal life from an initially diffuse projection. Disruption of N-methyl-D-aspartate (NMDA) receptor activity in the superior colliculus during this period interferes with map remodeling. In rats chronically treated with NMDA receptor antagonists during the first two postnatal weeks, aberrant axons remain and arborize at topographically incorrect sites. These results indicate that, at a stage preceding visually evoked activity, normal NMDA receptor function is important for the development of an ordered neural map in the mammalian brain.

2-Amino-5-phosphonovalerate↗

A novel myosin-like protein (myocilin) expressed in the connecting cilium of the photoreceptor: molecular cloning, tissue expression, and chromosomal mapping.

We have isolated a human cDNA clone encoding a novel acidic protein of MW 55,000 that we designated "myocilin" since it has homology to myosin and is localized preferentially in the ciliary rootlet and basal body of the connecting cilium of photoreceptor cells. The deduced amino acid sequence of human myocilin showed significant homologies with nonmuscle myosin of Dictyostelium discoideum in the N-terminal region and also with olfactomedin of bullfrog in the C-terminal region. Myocilin contained a leucine zipper-like motif similar to that seen in kinectin and other cytoskeletal proteins. These findings suggest that myocilin is a novel cytoskeletal protein involved in the morphogenesis of ciliated neuroepithelium such as photoreceptor cells. The myocilin gene (MYOC) was mapped to human chromosome 1q23-q24 by fluorescence in situ hybridization.

Amino Acid Sequence↗

A simple method for serving Web hypermaps with dynamic database drill-down.

BACKGROUND: HealthCyberMap http://healthcybermap.semanticweb.org aims at mapping parts of health information cyberspace in novel ways to deliver a semantically superior user experience. This is achieved through "intelligent" categorisation and interactive hypermedia visualisation of health resources using metadata, clinical codes and GIS. HealthCyberMap is an ArcView 3.1 project. WebView, the Internet extension to ArcView, publishes HealthCyberMap ArcView Views as Web client-side imagemaps. The basic WebView set-up does not support any GIS database connection, and published Web maps become disconnected from the original project. A dedicated Internet map server would be the best way to serve HealthCyberMap database-driven interactive Web maps, but is an expensive and complex solution to acquire, run and maintain. This paper describes HealthCyberMap simple, low-cost method for "patching" WebView to serve hypermaps with dynamic database drill-down functionality on the Web. RESULTS: The proposed solution is currently used for publishing HealthCyberMap GIS-generated navigational information maps on the Web while maintaining their links with the underlying resource metadata base. CONCLUSION: The authors believe their map serving approach as adopted in HealthCyberMap has been very successful, especially in cases when only map attribute data change without a corresponding effect on map appearance. It should be also possible to use the same solution to publish other interactive GIS-driven maps on the Web, e.g., maps of real world health problems.

Journal Article↗

Multimodal characterisation of cortical areas by multivariate analyses of receptor binding and connectivity data.

Cortical areas are regarded as fundamental structural and functional units within the information processing networks of the brain. Their properties have been described extensively by cyto-, myelo- and chemoarchitectonics, cortical and extracortical connectivity patterns, receptive field mapping, activation properties, lesion effects, and other structural and functional characteristics. Systematic integrative approaches aiming at multimodal characterisations of cortical areas or at the delineation of global features of the cortical network, however, are still scarce and usually limited to a single data modality, such as cytoarchitectonical or tract tracing data. Here we describe a methodological framework for the systematic evaluation, comparison and integration of different data modalities from the brain and demonstrate its practical application and significance in the analysis of receptor binding and connectivity data within the motor and visual cortices of macaque monkeys. The framework builds on algorithmic methods to convert data between different cortical parcellation schemes, as well as on statistical techniques for the exploration of multivariate data sets comprising data of different types and scales. Thereby, we establish a relationship between intrinsic area properties as expressed by quantitative receptor binding, and extrinsic inter-area communication, which relies on anatomical connectivity. Our analyses provide preliminary evidence for a good correspondence of these two data types in the motor cortex, and their partial discrepancy in the visual cortex, raising hypotheses about the different organisational aspects highlighted by receptors and connectivity. The methodological framework presented here is flexible enough to accommodate a wide range of further data modalities, and is specific enough to permit novel insights and predictions concerning brain organisation. Thus, this approach promises to be very useful in the endeavour to characterise multimodal structure-function relationships in the brain.

Animals↗