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Structure of poly d(AI).poly d(CT) in two different packing arrangements.

X-ray diffraction analysis of poly d(AI).poly d(CT) in oriented and polycrystalline fibers has revealed the DNA structure to be a 10-fold, right-handed, antiparallel, Watson-Crick base paired double helix in two distinct packing arrangements corresponding to one and two helices, respectively, in the unit cell. The helix pitch is 32.1 A and 32.4 A in the two cases, almost 1.5 A shorter than in classical B-DNA. The resulting B'-DNA geometry, described in terms of a tetranucleotide repeat which is conformationally similar to B-DNA, has its minor groove closely shut and major groove correspondingly widened, thus striking a sharp morphological contrast to B-DNA. According to difference electron density maps, a spine of hydration along the minor groove connects both strands and provides structural stability; ordered sodium ions and water molecules are actively involved in bridging the phosphate groups of neighboring helices. The crystallographic R-values for these two allomorphs are 0.26 and 0.20, respectively, for data up to 3.0 A resolution.

Crystallography, X-Ray↗

Purification and characterization of a 190-kD microtubule-associated protein from bovine adrenal cortex.

A heat-stable microtubule-associated protein (MAP) with molecular weight of 190,000, termed 190-kD MAP, was purified from bovine adrenal cortex. This MAP showed the same level of ability to promote tubulin polymerization as did MAP2 and tau from mammalian brains. Relatively high amounts of 190-kD MAP could bind to microtubules reconstituted in the presence of taxol. At maximum 1 mol of 190-kD MAP could bind to 2.3 mol of tubulin. 190-kD MAP was phosphorylated by a cAMP-dependent protein kinase prepared from sea urchin spermatozoa and by protein kinase(s) present in the microtubule protein fraction prepared from mammalian brains. The maximal numbers of incorporated phosphate were approximately 0.2 and approximately 0.4 mol per mole of 190-kD MAP, respectively. These values were lower than that of MAP2, which could be heavily phosphorylated by the endogenous protein kinase(s) up to 5 mol per mole of MAP2 under the same assay condition. 190-kD MAP had no effects on the low-shear viscosity of actin and did not induce an increase in turbidity of the actin solution. It was also revealed that 190-kD MAP does not cosediment with actin filaments. These data clearly show that, distinct from MAP2 and tau, this MAP does not interact with actin. Electron microscopic observation of the rotary-shadowed images of 190-kD MAP showed the molecular shape to be a long, thin, flexible rod with a contour length of approximately 100 nm. Quick-freeze, deep-etch replicas of the microtubules reconstituted from 190-kD MAP and brain tubulin revealed many cross-bridges connecting microtubules with each other.

Adrenal Cortex↗

The UK Human Genome Mapping Project online computing service.

This paper presents an overview of computing and networking facilities developed by the Medical Research Council to provide online computing support to the Human Genome Mapping Project (HGMP) in the UK. The facility is connected to a number of other computing facilities in various centres of genetics and molecular biology research excellence, either directly via high-speed links or through national and international wide-area networks. The paper describes the design and implementation of the current system, a 'client/server' network of Sun, IBM, DEC and Apple servers, gateways and workstations. A short outline of online computing services currently delivered by this system to the UK human genetics research community is also provided. More information about the services and their availability could be obtained by a direct approach to the UK HGMP-RC.

Computer Communication Networks↗

The columnar organization of the neocortex.

The modular organization of nervous systems is a widely documented principle of design for both vertebrate and invertebrate brains of which the columnar organization of the neocortex is an example. The classical cytoarchitectural areas of the neocortex are composed of smaller units, local neural circuits repeated iteratively within each area. Modules may vary in cell type and number, in internal and external connectivity, and in mode of neuronal processing between different large entities; within any single large entity they have a basic similarity of internal design and operation. Modules are most commonly grouped into entities by sets of dominating external connections. This unifying factor is most obvious for the heterotypical sensory and motor areas of the neocortex. Columnar defining factors in homotypical areas are generated, in part, within the cortex itself. The set of all modules composing such an entity may be fractionated into different modular subsets by different extrinsic connections. Linkages between them and subsets in other large entities form distributed systems. The neighborhood relations between connected subsets of modules in different entities result in nested distributed systems that serve distributed functions. A cortical area defined in classical cytoarchitectural terms may belong to more than one and sometimes to several distributed systems. Columns in cytoarchitectural areas located at some distance from one another, but with some common properties, may be linked by long-range, intracortical connections.

Animals↗

Rewiring the olfactory bulb: changes in odor maps following recovery from nerve transection.

Recent studies have shown that axons from olfactory receptor subtypes converge onto glomeruli in fixed positions within the olfactory bulb. Different receptor subtypes project to different glomeruli, forming a spatial distribution of odor information or 'odor maps'. Olfactory receptor neurons are continuously replaced throughout the life span of an animal, yet they preserve this highly localized mapping of receptor subtypes. In this study we used a transgenic mouse (P2-IRES-tau-lacZ) to map axons from a single receptor subtype (P2 receptors) in order to determine if regenerating axons were able to re-establish the P2 receptor map following nerve transection. Results confirm that P2 receptor axons retain their capacity to grow back to the olfactory bulb and converge onto glomeruli following nerve transection. However, the location and number of convergence sites was significantly altered compared to the control map. This change in the spatial distribution of axons alters the topography of odor mapping and has important implications for the processing of olfactory information. Findings from this study may explain why animals recovering from nerve injury require odor training before odor discrimination is restored. Future studies of olfactory receptor mapping could prove helpful in planning strategies to rewire connections in the brain and to restore function following injury or neurological disease.

Animals↗

Synthesis, restriction analysis, and molecular cloning of near full length DNA complementary to bovine parathyroid hormone mRNA.

DNA complementary (cDNA) to a partially purified preparation of bovine parathyroid hormone mRNA was synthesized using avian myeloblastosis viral reverse transcriptase. The PTH cDNA contained about 750 bases and was greater than 95% sensitive to digestion by S1 nuclease. Analysis of the mRNA preparation by excess RNA hybridization to the PTH cDNA revealed one rapidly hybridizing component consisting of 50% of the PTH cDNA. Sequential incubation of the PTH mRNA with reverse transcriptase and E. coli DNA polymerase I produced near full length double-stranded PTH cDNA. Of the 22 restriction endonucleases tested, double-stranded PTH cDNA could be cleaved with Alu I, Mbo II, Sau 3A, Sst I, and Taq I. The restriction fragments corresponding to the 5' terminus of the sense strand were identified for the last three enzymes by comparing the size of fragments obtained from PTH cDNA before and after cleavage of the hairpin loop connecting the two strands by S1 nuclease. The restriction map of the cDNA was used to detect clones of bacteria containing recombinant plasmids with near full length PTH cDNA inserts.

Animals↗

ASmodeler: gene modeling of alternative splicing from genomic alignment of mRNA, EST and protein sequences.

Alternative splicing is in important mechanism of modulating gene function and expression which greatly expands transcriptome diversity. ASmodeler is a novel web-based utility that finds gene models including alternative splicing events from genomic alignment of mRNA, EST and protein sequences. User-supplied sequences are aligned against the genome map using the BLAT and SIM4 programs. Resulting exon connectivity is analyzed by applying graph-theoretic methods to build all possible gene models including splice variants. The algorithm essentially combines the genome-based sequence clustering and transcript assembly procedures in a coherent fashion. In addition to the user-supplied sequences, UniGene clusters and many well-known gene predictions such as Genscan, Ensembl and Acembly may be included in gene modeling. The current implementation supports human, mouse and rat genomes. ASmodeler is available at http://genome.ewha.ac.kr/ECgene/ASmodeler/.

Algorithms↗

Core tracing: depicting connections between features in electron density.

Core tracing is a threshold-independent method of determining connectivity (long chains of high-density values) in electron-density maps. It gives visually sparse pictures of large volumes which are useful for initial fitting and for molecular-boundary determination. New methods for visual presentation of the traces are suggested by the way that the connectivity is parameterized in terms of local connections between maxima and the saddle (lowest) points along the connecting paths. The algorithm also partitions the density into small compact volumes containing the maxima. These volumes are useful for localization and statistical analysis.

Journal Article↗

Interaction network among Escherichia coli membrane proteins involved in cell division as revealed by bacterial two-hybrid analysis.

Formation of the Escherichia coli division septum is catalyzed by a number of essential proteins (named Fts) that assemble into a ring-like structure at the future division site. Several of these Fts proteins are intrinsic transmembrane proteins whose functions are largely unknown. Although these proteins appear to be recruited to the division site in a hierarchical order, the molecular interactions underlying the assembly of the cell division machinery remain mostly unspecified. In the present study, we used a bacterial two-hybrid system based on interaction-mediated reconstitution of a cyclic AMP (cAMP) signaling cascade to unravel the molecular basis of septum assembly by analyzing the protein interaction network among E. coli cell division proteins. Our results indicate that the Fts proteins are connected to one another through multiple interactions. A deletion mapping analysis carried out with two of these proteins, FtsQ and FtsI, revealed that different regions of the polypeptides are involved in their associations with their partners. Furthermore, we showed that the association between two Fts hybrid proteins could be modulated by the coexpression of a third Fts partner. Altogether, these data suggest that the cell division machinery assembly is driven by the cooperative association among the different Fts proteins to form a dynamic multiprotein structure at the septum site. In addition, our study shows that the cAMP-based two-hybrid system is particularly appropriate for analyzing molecular interactions between membrane proteins.

Adenylyl Cyclases↗

Plasticity and primary motor cortex.

One fundamental function of primary motor cortex (MI) is to control voluntary movements. Recent evidence suggests that this role emerges from distributed networks rather than discrete representations and that in adult mammals these networks are capable of modification. Neuronal recordings and activation patterns revealed with neuroimaging methods have shown considerable plasticity of MI representations and cell properties following pathological or traumatic changes and in relation to everyday experience, including motor-skill learning and cognitive motor actions. The intrinsic horizontal neuronal connections in MI are a strong candidate substrate for map reorganization: They interconnect large regions of MI, they show activity-dependent plasticity, and they modify in association with skill learning. These findings suggest that MI cortex is not simply a static motor control structure. It also contains a dynamic substrate that participates in motor learning and possibly in cognitive events as well.

Animals↗

NOGA-guided analysis of regional myocardial perfusion abnormalities treated with intramyocardial injections of plasmid encoding vascular endothelial growth factor A-165 in patients with chronic myocardial ischemia: subanalysis of the EUROINJECT-ONE multicenter double-blind randomized study.

BACKGROUND: The aim of this substudy of the EUROINJECT-ONE double-blind randomized trial was to analyze changes in myocardial perfusion in NOGA-defined regions with intramyocardial injections of plasmid encoding plasmid human (ph)VEGF-A(165) using an elaborated transformation algorithm. METHODS AND RESULTS: After randomization, 80 no-option patients received either active, phVEGF-A165 (n=40), or placebo plasmid (n=40) percutaneously via NOGA-Myostar injections. The injected area (region of interest, ROI) was delineated as a best polygon by connecting of the injection points marked on NOGA polar maps. The ROI was projected onto the baseline and follow-up rest and stress polar maps of the 99m-Tc-sestamibi/tetrofosmin single-photon emission computed tomography scintigraphy calculating the extent and severity (expressed as the mean normalized tracer uptake) of the ROI automatically. The extents of the ROI were similar in the VEGF and placebo groups (19.4+/-4.2% versus 21.5+/-5.4% of entire myocardium). No differences were found between VEGF and placebo groups at baseline with regard to the perfusion defect severity (rest: 69+/-11.7% versus 68.7+/-13.3%; stress: 63+/-13.3% versus 62.6+/-13.6%; and reversibility: 6.0+/-7.7% versus 6.7+/-9.0%). At follow-up, a trend toward improvement in perfusion defect severity at stress was observed in VEGF group as compared with placebo (68.5+/-11.9% versus 62.5+/-13.5%, P=0.072) without reaching normal values. The reversibility of the ROI decreased significantly at follow-up in VEGF group as compared with the placebo group (1.2+/-9.0% versus 7.1+/-9.0%, P=0.016). Twenty-one patients in VEGF and 8 patients in placebo group (P<0.01) exhibited an improvement in tracer uptake during stress, defined as a >or =5% increase in the normalized tracer uptake of the ROI. CONCLUSIONS: Projection of the NOGA-guided injection area onto the single-photon emission computed tomography polar maps permits quantitative evaluation of myocardial perfusion in regions treated with angiogenic substances. Injections of phVEGF A165 plasmid improve, but do not normalize, the stress-induced perfusion abnormalities.

Algorithms↗

Analysis on the dispersal pattern of newly introduced Latrodectus hasseltii (Araneae: Theridiadae) in Japan by spider diagram.

The dispersal pattern of the widow spider Latrodectus hasseltii (Araneae: Theridiadae), in Osaka Prefecture, Japan, was analyzed from accumulated survey data of spider distribution. All confirmed infested points from 1996 to 2002 were plotted on a map, and infested points in each survey year were connected with the nearest previously confirmed points. Expansion patterns of the confirmed habitats were analyzed by year and dispersion distances were classified by using spider diagram, one of method of geographical information system spatial analysis. L. hasseltii moves by walking step by step on the ground, mainly in coastal areas during early stages of infestation, and also can be conveyed over distances by various vehicles, mainly in inland areas. These analyses show L. hasseltii does not move by ballooning.

Animals↗

Topographic analysis with monoclonal anti-idiotopes: probing the functional anatomy of immunoglobulin variable domains.

Attempts to correlate immunoglobulin variable domain functional properties with variable domain primary structure have been valuable, but these efforts have suggested that to more fully account for variable domain function in terms of structure will require knowledge of molecular relationships in three dimensions. In this review we describe generally applicable methods, using monoclonal anti-idiotopes, for the determination of spatial relationships of idiotopes relative to one another and relative to two orienting structural markers of variable domains: 1) the hapten-binding site and 2) the junction of the variable and constant domains. Using these methods it has been possible to construct an oriented idiotope map which spans the variable domain along an axis connecting the paratope and the variable domain-constant domain junction. In addition, it has been possible to correlate idiotope position with other properties of idiotope expression. This approach may contribute to the development of predictive principles of idiotope expression.

Animals↗

[Theory and practical approaches in catheter ablation of atrial flutter].

Atrial flutter can be understood as atrial tachycardia due to a single intraatrial macroreentrant circuit that is determined by fixed or functional boundaries. In various types of atrial flutter, radiofrequency ablation became an established curative therapy. During the course of an ablation procedure, initially, the reentrant circuit has to be determined, e.g. by activation and entrainment mapping. Subsequently, the boundaries have to be identified. By connecting two appropriate boundaries with a linear lesion, the intraatrial reentrant circuit can be inhibited. Finally, it should be proven that the linear lesion results in a complete line of conduction block. Doing so, the acute and long-term results of atrial flutter ablation are comparable to those of other supraventricular tachycardias.

Atrial Flutter↗

[Temporal organization of interneuronal relations in the cerebral cortex of the cat in relation to the level of alimentary motivation].

By means of records of multicellular activity, interneuronal relations and their modifications in two cortical zones (Visual and motor) were studied in cats at different levels of alimentary motivation. For quantitative evaluation of interneuronal relations the statistic method of cross-correlation analysis of impulse trains was used in determining the probability of the appearance of the discharge of one neurone after the impulse of the other one. For groups of neurones in both investigated cortical areas, three-neurones microsystems were singled out and their activity was analyzed by temporal parameters of interaction between neurones at the interval of 120 ms, both within one microarea (intraanalyzer connections) and between microareas of two cortical zones. The correlation of temporal parameters of interneuronal connections (temporal delays in the activity of neuronal pairs) changed depending on spatial localization of neurones and functional condition of the animals. The existence is suggested of "informational" (1-30 ms) and "motivational" (90-120 ms) values of interneuronal relations for interanalyser connections.

Animals↗

[Functional interactions between the rabbit hypothalamus and neocortex].

In acute experiments on rabbits under chloralose-nembutal anaesthesia, studies have been made on the distribution and the pattern of the evoked potentials (EP) in neocortical structures of the brain cortex during electrical stimulation of different hypothalamic nuclei. It was found that during stimulation of the supramammilar area of the hypothalamus, the focus of maximum activity of cortical responses of the negative-positive configuration is localised in the median parts of precentral agranular region. During stimulation of the posterior hypothalamic area, the EP were of the positive-negative-positive type. Registration of the EP in different cortical layers during stimulation of the posterior hypothlamic nuclei showed that from 100 to 100 micrograms the amplitude of EP components decreases, whereas at the level of 600 mu the inversion of the EP takes place. At the depth of 1.800 mu the EP elicited by stimulation of the posterior hypothalamus had a form of a positive oscillation with a long latent period. It is concluded that the posterior hypothalamic nuclei have a vast representation in rabbit neocortex, this representation being realized by both polysynaptic and olygosynaptic systems of connections.

Animals↗

Detection and epitope analysis of autoantigen-reactive T cells to the U1-small nuclear ribonucleoprotein A protein in autoimmune disease patients.

T cells that recognize autoantigens might play an important role in autoimmune diseases. To analyze "autoantigen-reactive T cell" in patients with an autoimmune disease, a human lymphocyte proliferation assay using soluble recombinant autoantigens has been conducted. PBMC from mixed connective tissue disease and SLE patients who possessed anti-U1-small nuclear ribonucleoprotein (snRNP) A autoantibodies responded to a recombinant U1-snRNP A protein. In contrast, PBMC from healthy subjects or autoimmune disease patients not possessing anti-U1-snRNP A autoantibodies did not show such responses. In addition, this proliferative response was inhibited by anti-CD4 mAb. Limiting dilution analyses revealed that the autoantigen-reactive T cells exist in relatively high frequency (1 of 4,065 to 1 of 23,256) in the mixed connective tissue disease patients. Moreover, by T cell epitope mapping, the T cell epitope area was found to be located in the C-terminus of the U1-snRNP A protein, overlapping the B cell epitope area that has been reported. These findings suggest the presence of autoantigen-reactive T cells in such patients, and when these T cells are activated, they may play a central role in the autoantibody generation.

Amino Acid Sequence↗

Influence of the tectal zone on the distribution of synaptic boutons in the brainstem of goldfish.

This study investigated whether the topographic differences in the functional properties of the tectal motor map of goldfish are related to particular patterns of connections with downstream structures. With this aim, the distribution of synaptic boutons in the mesencephalic and rhombencephalic structures was studied after discrete injections of the tracer biotinylated dextran amine were placed at separate sites along the tectal anteroposterior axis. Irrespective of the location of the injection site, the boutons were more abundant in the mesencephalon than in the rhombencephalon, and they were located chiefly ipsilaterally all throughout the brainstem. In the mesencephalon, the boutons were found in its ventrolateral reticular formation and, to a lesser extent, in the nucleus of the medial longitudinal fasciculus, the oculomotor and isthmi nuclei, and the torus semicircularis. In the mesencephalic reticular formation, the bouton location was distributed topographically with respect to the injection site. Terminals were also observed in the nucleus of the medial longitudinal fasciculus after injections into anteromedial or middle tectal zones. In the oculomotor nucleus, boutons were present exclusively in the case of the anteromedial injection. In the rhombencephalon, most boutons were found in the superior reticular formation, and their number decreased in the medial and inferior reticular formations. A topographic distribution could be observed within the superior reticular formation, although its density was attenuated compared with that observed in the mesencephalic reticular formation. The domains of synaptic endings on the ipsilateral side were different from those on the contralateral side: The ipsilateral synaptic endings were located more medially. Finally, a few boutons were also found in the vestibulocerebellar area on either the ipsilateral or the contralateral side, depending on the injection site. From these data, the authors conclude that, in goldfish, irrespective of the tectal injection site, the endings are in similar nuclei in the brainstem; however, the distribution of synaptic boutons within such nuclei can be related to the functional properties of each tectal zone.

Anatomy, Artistic↗