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Metal-induced tautomerization of p- to o-quinone compounds: experimental evidence from CuI and ReI complexes of azophenine and DFT studies.

Azophenine (7,8-diphenyl-2,5-bis(phenylamino)-p-quinonediimine, L(p)) reacts with [Cu(PPh3)4](BF4) or [Re(CO)(5)Cl] to yield the (Ph3P)(2)Cu(+) or [(OC)(3)ClRe] complex of the tautomeric form 7,8-diphenyl-4,5-bis(phenylamino)-o-quinonediimine, L(o), as evident from structure determinations and from very intense metal-to-ligand charge transfer (MLCT) transitions in the visible region. Time-dependent DFT (TD-DFT) calculations on model complexes [(N intersection N)Re(CO)(3)Cl] confirm the spectroscopic results, showing considerably higher oscillator strengths of the MLCT transition for the o-quinonediimine complexes in comparison to compounds with N intersection N=1,4-dialkyl-1,4-diazabutadiene. The complexes are additionally stabilized through hydrogen bonding between two now ortho-positioned NHPh substituents and one fluoride of the BF(4) (-) anion (Cu complex) or the chloride ligand (Re complex). DFT Calculations on the model ligands p-quinonediimine or 2,5-diamino-p-quinonediimine and their ortho-quinonoid forms with and without Li(+) or Cu(+) are presented to discuss the relevance for metal-dependent quinoproteins.

Journal Article↗

Anion control over interpenetration and framework topology in coordination networks based on homoleptic six-connected scandium nodes.

Reaction of ScX3 (X=NO3-, CF3SO3-, ClO4-) with 4,4'-bipyridine-N,N'-dioxide (L) affords topologically distinct six-connected three-dimensional coordination frameworks, {[Sc(L)3](NO3)3}(infinity) (1), {[Sc(L)3](CF3)SO3)3(CH3OH)2.7(H2O)3}(infinity) (2), {[Sc(L)3](ClO4)3}(infinity) (3) and {[Sc(L)4(H2O)2](ClO4)3}(infinity) (4). Compounds 1, 2 and 3 are networks based on octahedrally co-ordinated ScO6 centres bound through six oxygen atoms from six separate N-oxide ligands L. Compounds 1 and 3 are doubly interpenetrated and have alpha-polonium-type structures of 4(12)6(3) topology based upon three intersecting (4,4) nets. The structure of 2 is unusual and shows parallel, co-planar layers of (4,4) nets connected in a criss-crossed fashion to afford a new 4(8)6(6)8 topology. In 4 only four ligands L bind to each Sc(III) centre with two additional water molecules bridging metal nodes. Significantly, the bridges formed by L do not sit in a plane and if connections through L are considered alone the resultant structure is a diamondoid array typically based upon a tetrahedral connecting node at Sc. Five interpenetrating diamondoid networks are observed that are cross-bridged by water molecules to form a single three-dimensional array of 4(8)6(7) topology. Compound 4 can also be viewed as incorporating two intersecting (4,4) grids based upon two ligands L and two bridging waters. Thus, variation of anion, solvent and conditions critically affects the structures of products formed, and the series of polymers reported herein illustrates how tectons based upon (4,4) grids can be combined and distorted to form non-NaCl topologies and even cross-bridged, multiply interpenetrated diamondoid materials. Both compounds 2 and 4 represent unusual examples of self-penetrated coordination frameworks.

Journal Article↗

Monoamine-accumulating ganglion cell type of the cat's retina.

A monoamine-accumulating ganglion cell type has been identified in an in vitro preparation of the cat's retina by a catecholamine-like fluorescence that appears following intravitreal injections of dopamine and the indoleaminergic transmitter analog, 5,7-dihydroxytryptamine (5,7-DHT). A subpopulation of large, weakly fluorescing neurons were identified as composing a single, morphologically distinct ganglion cell type by intracellular injections of horseradish peroxidase (HRP). In a sample of 374 HRP-filled cells soma diameter ranged from 13-21 microns (mean +/- SD = 16.6 +/- 1.3). Dendritic field size increased with increasing retinal eccentricity from 150-200 microns diameter at 0.5 mm from the area centralis to 600-800 microns diameter in the far retinal periphery. Dendrites are thin (approximately 1 micron diameter), show a characteristic branching pattern, and are narrowly stratified at the outer border of the inner plexiform layer. The monoamine-accumulating ganglion cell and the outer (OFF-center) alpha cell occupy distinct strata within sublamina a of the inner plexiform layer separated by a gap of about 5 microns. The total number of monoamine-accumulating (MA) ganglion cells was estimated at 5,400, about 3.5% of the total ganglion cell population. Spatial density of the MA ganglion cells, calculated from cell counts made in vitro, ranges from 60 cells/mm2 near the area centralis to 5 cells/mm2 in the far retinal periphery. A coverage factor (density x dendritic field area) of 2.2 was maintained from central to peripheral retina. The nature of the dendritic overlap was observed directly by making HRP injections into several neighboring ganglion cells. Five to seven neighboring dendritic trees extensively overlapped a given cell's dendritic field. However the dendritic processes did not intersect randomly but tended to interdigitate such that a uniform interdendritic spacing and density of dendritic processes was constructed locally within the dendritic plexus. Rotation of individual dendritic trees from their normal orientation produced a dramatic 4-5-fold increase in the number of dendritic intersections, suggesting that an active, local mechanism operates in the precise placement of individual dendrites within the plexus. The monoamine-accumulating ganglion cell appears morphologically equivalent to the delta ganglion cell (Boycott and Wässle; J. Physiol. (Lond.) 240:397-419, '74; Kolb et al.; Vision Res. 21:1081-1114, '81) and to the recently recognized indoleamine-accumulating ganglion cell (Wässle et al: J. Neurosci. 7:1574-1585, '87).(ABSTRACT TRUNCATED AT 400 WORDS)

5,7-Dihydroxytryptamine↗

High resolution optics combined with high spatial reproducibility in flow.

Accurate sizing in flow using optical methods generally requires high resolution optics and specially designed flow systems. Flow systems developed by this group have following features: (a) double sheath configuration for optical index match, (b) no curved optical surface in the sensing area, (c) gradual hydrodynamic focusing over a long distance to minimize mechanical shearing, (d) precision spatial positioning of cells by reducing suspension fluid diameter to a cell diameter or less, (e) total thickness between outer surfaces of the flow chamber at the viewing area of 1.5 mm or less. Cells intersect a laser light beam focussed go circular as well as elliptical cross-sections or 1 micron or less in diameter. Cellular extinction is monitored during transit through the beam. Cell length is derived from the time for flight measurement and corrected for absolute values by continuous velocity reference using a second laser beam intersecting the cell stream at a predetermined distance. This second spot may be circular or elliptical, of a different polarization and/or frequency. Simultaneous fluorescence intensity and diameter measurements were performed on test particles using different optical geometries. The influence of the particle structure on fluorescence measurements is demonstrated where high resolution sizing is required at the same time.

Cytological Techniques↗

Selective control of neuronal cluster size at the forebrain/midbrain boundary by signaling from the prechordal plate.

Within the vertebrate embryonic neural plate, the first neuronal clusters often differentiate at the border of patterning identities. Whether the information inherent in the intersection of patterning identities alone controls all aspects of neuronal cluster development (location, identity, and size) is unknown. Here, we focus on the cluster of the medial longitudinal fascicle (nMLF) and posterior commissure (nPC), located at the forebrain/midbrain (fore/mid) boundary, to address this issue. We first identify expression of the transcription factor Six3 as a common and distinct molecular signature of nMLF and nPC neurons in zebrafish, and we use this marker to monitor mechanisms controlling the location and number of nMLF/nPC neurons. We demonstrate that six3 expression is induced at the fore/mid boundary in pax2.1/no-isthmus and smoothened/slow muscle omitted mutants, where identities adjacent to the six3 cluster are altered; however, in these mutants, the subpopulation of six3-positive cells located within the mispatterned territory is reduced. These results show that induction of the six3 cluster is triggered by the information derived from the intersection in patterning identities alone, whereas correct cluster size depends, in a modular manner, on the identities themselves. The size of the six3 cluster is also controlled independently of neural tube patterning: we demonstrate that the prechordal plate (PCP) is impaired in mixer/bonnie and clyde mutants and that this phenotype secondarily results in an increased production of six3-positive cells at the fore/mid boundary, without correlatively affecting patterning in this area. Thus, a signaling process originating from the PCP distinguishes between neural patterning and the control of six3 cluster size at the fore/mid junction in vivo. Together, our results suggest that a combination of patterning-related and -unrelated mechanisms specifically controls the size of individual early neuronal clusters within the anterior neural plate.

Animals↗

Ferguson plots based on absolute mobilities in polyacrylamide gel electrophoresis: dependence of linearity of polymerization conditions and application to the determination of free mobility.

In contrast to Ferguson plots based on relative mobilities, Ferguson plots of proteins in polyacrylamide gel electrophoresis based on their absolute mobilities were found to be linear under unusual polymerization conditions which yield relatively wide gel fibers and a low total fiber length per unit weight, but not under previously and commonly used conditions. These linear Ferguson plots in gels of 1, 3 and 5% crosslinking intersect at a single gel concentration between 1 and 2% T (M-point). It is postulated that the measure of free mobility of the proteins is the M-point, and not the intercept of their Ferguson plots with the mobility axis as assumed previously. This postulate abolishes the well-known paradoxical interpretation of the increase with %C of the linearly extrapolated intercept of the Ferguson plot with the log(mobility) axis (designated Yo) in terms of free mobility. The postulate is also compatible with the interpretation of the points of intersection of the Ferguson plots of oligomeric series of proteins at finite gel concentrations (designated mu-points) as their common free mobilities.

Buffers↗

Information on DNA conformation derived from transverse pore gradient gel electrophoresis in conjunction with an advanced data analysis applied to capillary electrophoresis in polymer media.

Abnormally slow migration of DNA is conventionally viewed as being due to an abnormal conformation relative to "linear" standards. The evidence for this rests on a few instances where nonlinear DNA structures have been established by independent methods and yield low mobilities relative to standards. Transverse pore gradient gel electrophoresis of authentically bent kinetoplast DNA and of an upstream activator sequence (UAS) of an E. coli operon promoter shows in addition that curves of migration distance vs. gel concentration ("Ferguson curves") of such abnormally conformed DNA differ from those of "linear" standards. Since Ferguson curves are interpretable with regard to molecular size in concordance with a mathematical model (Ogston model), transverse pore gradient gel electrophoresis provides a simple means of correlating abnormally slow migration of DNA with molecular size. In addition, transverse pore gradient gel electrophoresis is able to distinguish between DNA banding which exhibits a steeper dependence on gel concentration than "linear" standards from one which shows the same dependence. The former appears characteristic of circularly bent DNA and gives rise to a substantial retardation, the latter of bending across a knot or kink in the DNA chain associated with a relatively minor retardation relative to standards. Circularly bent restriction fragments formed from kinetoplast DNA retain the characteristic intersecting Ferguson curves on the transverse pore gradient gel. Another authentically "abnormal" DNA structure recognizable on transverse pore gradient gels is supercoiled DNA derived from the reaction of topoisomerase with a plasmid. Different lengths of supercoiled sequences give rise to parallel Ferguson curves clearly intersecting with those of linear standards.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Focused electrophoretic motion and selected electrokinetic dispensing of particles and cells in cross-microchannels.

The electrokinetic focusing and the resultant accelerated electrophoretic motion of polystyrene particles and red blood cells were visualized in microfluidic cross-channels. The experimentally measured width of the focused stream and the measured velocity increase of particles and cells at different voltage ratios follow the proposed analytical formula within the experimental error. The attained velocity increase is insensitive to the particle size, particle property (i.e., particle or cell), and particle trajectory. By solving the electrical potential field in the cross-channel at the experimental conditions, we demonstrate that the squeezed electrical field lines in the channel intersection determine the shape of the focused stream, and the nonuniform distribution of axial electrical field strength underlies the variation of particle/cell electrophoretic velocity through the focusing region. However, the dielectrophoretic force resulting from the nonuniform electrical field in the intersection seems to push the acceleration region of particles and cells slightly in the downstream direction. We have also achieved the single particle/cell dispensing by instantly triggering an electrical pulse perpendicular to the focused particulate flow in a double-cross microchannel. The electrokinetic manipulation of particle/cell in microchannels demonstrated in this work can be used for developing integrated lab-on-a-chip devices for studies of cells.

Animals↗

Numerical analysis of an electrokinetic double-focusing injection technique for microchip CE.

The injection techniques in electrophoresis microchips play an important role in the sample-handling process, whose characteristics determine the separation performance achieved, and the shape of a sample plug delivered into the separation channel has a great impact on the high-quality separation performance as well. This paper describes a numerical investigation of different electrokinetic injection techniques to deliver a sample plug within electrophoresis microchips. A novel double-focusing injection system is designed and fabricated, which involves four accessory arm channels in which symmetrical focusing potentials are loaded to form a unique parallel electric field distribution in the intersection of injection channel and separation channel. The parallel electric field effectuates virtual walls to confine the spreading of a sample plug at the intersection and prevents sample leakage into separation channel during the dispensing step. The key features of this technique over other injection techniques are the abilities to generate regular and nondistorted shape of sample plugs and deliver the variable-volume sample plugs by electrokinetic focusing. The detection peak in the proposed injection system is uniform regardless of the position of the detection probe in the separation channel, and the peak resolution is greatly enhanced. Finally, the double-focusing injection technique shows the flexibility in detection position and ensures improved signal sensitivity with good peak resolution due to the delivered high-quality sample plug.

Computer Simulation↗

Visual search and urban driving under the influence of marijuana and alcohol.

The purpose of the present study was to assess the effects of low doses of marijuana and alcohol, and their combination, on visual search at intersections and on general driving proficiency in the City Driving Test. Sixteen recreational users of alcohol and marijuana (eight males and eight females) were treated with these substances or placebo according to a balanced, 4-way, cross-over, observer- and subject-blind design. On separate evenings, subjects received weight-calibrated doses of THC, alcohol or placebo in each of the following treatment conditions: alcohol placebo + THC placebo, alcohol + THC placebo, THC 100 &mgr;g/kg + alcohol placebo, THC 100 &mgr;g/kg + alcohol. Alcohol doses administered were sufficient for achieving a blood alcohol concentration (BAC) of about 0.05 g/dl. Initial drinking preceded smoking by one hour. The City Driving Test commenced 15 minutes after smoking and lasted 45 minutes. The test was conducted over a fixed route within the city limits of Maastricht. An eye movement recording system was mounted on each subject's head for providing relative frequency measures of appropriate visual search at intersections. General driving quality was rated by a licensed driving instructor on a shortened version of the Royal Dutch Tourist Association's Driving Proficiency Test. After placebo treatment subjects searched for traffic approaching from side streets on the right in 84% of all cases. Visual search frequency in these subjects did not change when they were treated with alcohol or marijuana alone. However, when treated with the combination of alcohol and marijuana, the frequency of visual search dropped by 3%. Performance as rated on the Driving Proficiency Scale did not differ between treatments. It was concluded that the effects of low doses of THC (100 &mgr;g/kg) and alcohol (BAC < 0.05 g/dl) on higher-level driving skills as measured in the present study are minimal. Copyright 2001 John Wiley & Sons, Ltd.

Journal Article↗

Deregulation of cyclin E meets dysfunction in p53: closing the escape hatch on breast cancer.

In this review, we focus on pathways intersecting through p53 and cyclin E, highlighting how oncogenic effects of cyclin E deregulation, especially overexpression of shortened or low molecular weight (LMW) forms of cyclin E protein, are amplified by loss of regulatory control through p53 to promote tumor development. Expression of cyclin E protein promotes progression into S-phase, an activity opposed by p53-regulated activation of checkpoint controls or apoptosis. Loss of p53 function is an escape hatch by which tumor cells, initiated by a number of means including cyclin E deregulation, can avoid cell cycle arrest or cell death and progress through further stages of unchecked deregulation and growth. To determine how this escape hatch is opened and, ultimately, how to close it, we must understand the networks of normal signaling and processing in a cell and where they intersect.

Animals↗

Identification of fibers at risk for degeneration by diffusion tractography in patients at high risk for MS after a clinically isolated syndrome.

PURPOSE: Focal inflammatory/demyelinating lesions are thought to be the source of Wallerian degeneration or other injury to local, transiting fiber tracts in the brain or spinal cord in multiple sclerosis (MS). A methodology is established to isolate connections between focal demyelinating lesions and intersecting fibers to permit explicit analyses of the pathology of secondary fiber injury distant from the focal lesion. MATERIALS AND METHODS: A strategy is described and feasibility demonstrated in three patients with a clinically isolated syndrome and positive MRI (at high risk for MS). The strategy utilizes streamtube diffusion tractography to identify neuronal fibers that intersect a focal lesion and pass through a region of interest, in this case the corpus callosum, where distal (to focal lesion) interrogation can be accomplished. RESULTS: A sizeable fraction of the normal appearing white matter (NAWM) in the early stages of disease can be defined in the corpus callosum, which is distinctive in that this tissue connects to distant demyelinating lesions. CONCLUSION: The new class of tissue called fibers-at-risk for degeneration (FAR) can be identified and interrogated by a variety of quantitative MRI methodologies to better understand neuronal degeneration in MS.

Adult↗

Compliance of hippocampal neurons to patterned substrate networks.

Neuronal growth can be controlled in vitro by plating cells at low density and by differential adhesion between the cell and substrate. Primary cultures of rat hippocampal neurons were grown in serum-free culture on polylysine-coated glass coverslips patterned by selective laser ablation so as to leave grids of polylysine with varying linewidths (3, 5, and 10 microns), intersection distance (80, 120, and 160 microns), and nodal (intersection) diameter (5, 10, and 20 microns). Not only did somae strongly prefer the unablated polylysine areas, but they also migrated to loci where the local area of unablated polylysine was higher. These loci were the nodes, as opposed to the narrow connecting paths, and larger nodes, as compared with smaller nodes. Maximum migration to nodes of 88% occurred for a combination of 5-microns path width, 20-microns node diameter, and 80-microns path length. Daily observations indicated active migration to larger adhesive areas, which explains the differential compliance.

Animals↗

Three-dimensional anatomy of the cancellous structures within the proximal femur from computed tomography data.

Quantitative computed tomography was used to investigate the three-dimensional architecture and the density distribution of the cancellous structures of the proximal femur. We examined 10 femora from the cadavera of 10 individuals, 47-83 years old at the time of death. Three anatomic elements could be distinguished: the epiphysis, the epiphyseal scar, and the metaphysis. Although these elements constitute a functional unit, their individual cancellous patterns revealed significant structural differences. The epiphyseal segment had a more reticulate cancellous appearance, whereas the metaphysis demonstrated a more longitudinally oriented trabecular distribution. The three-dimensional reconstruction of the densest epiphyseal trabecular groups showed two different patterns: a dumbbell type (found in eight bones) and a hemispheric type (found in two bones). The epiphyseal scar was a clearly recognizable high-density structure found in all 10 bones. The epiphyseal scar-diaphysis angle was 13-26 degrees higher than the center column-diaphysis angle. The scar appeared as a tray supporting the epiphyseal cancellous structures, supported itself by the metaphyseal intersection of the main compressive and the arcuate trabecular systems. This intersection always occurred in a relatively small band-shaped zone under the central third of the epiphyseal scar. These three discrete anatomical segments within the proximal femur may reflect developmental and functional adaptations determined by joint incongruity or eccentric muscular activity. The present data will serve as a reference for future studies in which the cancellous patterns are used to help with the early diagnosis of states of disease.

Aged↗

Density compensation functions for spiral MRI.

In interleaved spiral MRI, an object's Fourier transform is sampled along a set of curved trajectories in the spatial frequency domain (k-space). An image of the object is then reconstructed, usually by interpolating the sampled Fourier data onto a Cartesian grid and applying the fast Fourier transform (FFT) algorithm. To obtain accurate results, it is necessary to account for the nonuniform density with which k-space is sampled. An analytic density compensation function (DCF) for spiral MRI, based on the Jacobian determinant for the transformation between Cartesian coordinates and the spiral sampling parameters of time and interleaf rotation angle, is derived in this paper, and the reconstruction accuracy achieved using this function is compared with that obtained using several previously published expressions. Various nonideal conditions, including intersecting trajectories, are considered. The new DCF eliminated intensity cupping that was encountered in images reconstructed with other functions, and significantly reduced the level of artifact observed when unevenly spaced sampling trajectories, such as those achieved with trapezoidal gradient waveforms, were employed. Modified forms of this function were found to provide similar improvements when intersecting trajectories made the spiral-Cartesian transformation noninvertible, and when the shape of the spiral trajectory varied between interleaves.

Humans↗

Mapping complex tissue architecture with diffusion spectrum magnetic resonance imaging.

Methods are presented to map complex fiber architectures in tissues by imaging the 3D spectra of tissue water diffusion with MR. First, theoretical considerations show why and under what conditions diffusion contrast is positive. Using this result, spin displacement spectra that are conventionally phase-encoded can be accurately reconstructed by a Fourier transform of the measured signal's modulus. Second, studies of in vitro and in vivo samples demonstrate correspondence between the orientational maxima of the diffusion spectrum and those of the fiber orientation density at each location. In specimens with complex muscular tissue, such as the tongue, diffusion spectrum images show characteristic local heterogeneities of fiber architectures, including angular dispersion and intersection. Cerebral diffusion spectra acquired in normal human subjects resolve known white matter tracts and tract intersections. Finally, the relation between the presented model-free imaging technique and other available diffusion MRI schemes is discussed.

Algorithms↗

Podophyllum hexandrum prevents radiation-induced neuronal damage in postnatal rats exposed in utero.

Podophyllum hexandrum has been shown to mitigate radiation injuries and especially the haemopoietic syndrome in adult mice. To monitor the radiation-induced changes in the nervous system, the neurons of postnatal young mice and their modification by P. hexandrum, were studied histologically for differences in the apical and basal dendritic branching and intersections in the CA1 neurons of the hippocampal region of rats which were delivered a 2 Gy gamma dose while in utero (day 17 of gestation). Irradiation significantly reduced the dendritic branching and intersections but pre-irradiation administration of the extract of P. hexandrum (i.p. 200 mg/kg/b.w., 2 h) reduced the damage in postnatal young mice. These studies indicate that P. hexandrum provides protection to neurons against radiation-induced damage and the mechanism of neuronal damage and its repair need to be investigated further.

Animals↗

Inhibition Effects in the Hydrolysis Reactions of Esters and Peptides Catalyzed by Carboxypeptidase A: An Example of Cooperative Binding Effects with a Monomeric Enzyme.

N-benzoyl-L-phenylalanyl-L-phenylalanine is an excellent peptide substrate for carboxy-peptidase A; at 30 degrees C and pH 7.5, K(m) is 2.6 x 10(-5) M while k(cat) is 177 s(-1) (k(cat)/K(m) = 6.8 x 10(6) M(-1) s(-1)). Indole-3-acetic acid is a noncompetitive or mixed inhibitor towards the peptide and toward hippuryl-L-phenylalanine; plots of E/V vs [Inhibitor] are linear. N-Benzoyl-L-phenylalanine is a competitive inhibitor of peptide hydrolysis, and plots of E/V vs [Inhibitor] are again linear. One molecule of inhibitor binds per active site, and these inhibitors bind in different sites. At constant peptide substrate concentration and a series of constant concentrations of indole-3-acetic acid, plots of E/V vs the concentration of N-benzoyl-L-phenylalanine are linear and intersect behind the E/V axis and above the [Inhibitor] axis. This shows that both inhibitors can bind simultaneously and that binding of one facilitates the binding of the other (beta = 0.18). Employing the ester substrate hippuryl-DL,beta-phenyllactate, the same type of behavior is observed in the reverse sense; N-benzoyl-L-phenylalanine is a linear noncompetitive inhibitor and indole-3-acetic acid is a linear competitive inhibitor. Again the two inhibitor plot is linear and intersects above the [Inhibitor] axis (beta = 0.12). Previous X-ray crystallographic studies have indicated that indole-3-acetic acid binds in the hydrophobic pocket of the S'(1) site, while N-benzoyl-L-phenylalanine binds in the S(1)-S(2) site. The product complex for hydrolysis of N-benzoyl-L-phenylalanyl-L-phenylalanine (phenylalanine + N-benzoyl-L-phenylalanine) occupies both of these sites. However, the present work shows that the peptide substrate does not bind to the enzyme at pH 7.5 so as to be competitive with indole-3-acetic acid. The binding sites may be formed via conformational changes induced or stabilized by substrate and product binding. Copyright 2000 Academic Press.

Journal Article↗