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Dendritic morphology and development in the ferret medial superior olivary nucleus.

Dendritic morphology and development in the medial superior olivary nucleus of the ferret were studied using the Golgi method. In the adult ferret most medial superior olivary neurons had disk-shaped dendritic fields. These dendritic fields were oriented such that cells in a coronal plane of section appeared bipolar with major lateral and medial dendritic axes. In the horizontal plane the dendrites radiated about the soma. Dendrites of principal cells branched distally into tufts of numerous, tertiary processes that were beaded and thin. Peripheral cells in the fiber mantle encircling the nucleus were generally spindle-shaped or tripolar and lacked the tufted dendrites of principal cells. The dendrites of these peripheral cells coursed parallel to the nucleus both dorsoventrally and rostrocaudally. Horizontally oriented dendrites were observed even at birth for some cells in the medial superior olivary nucleus and bipolar dendritic fields were typical of most cells by the end of the second postnatal week. Dendrites of immature cells varied in caliber and radiated in all directions from the soma. Around postnatal days 8-10 transient appendages appeared on the soma and dendrites, first proximally and then more distally. These appendages persisted until the first postnatal month. Cell size and dendritic radius increased markedly during this same period. Postnatal days 28-30 were marked by the first appearance of tufts of tertiary dendritic branches. The tendril-like processes continued to increase in length until about the end of the second postnatal month.

Aging↗

[Vector propagation algorithm based ECG simulation of bundle branch block].

The simulation of excitation propagation's process in human heart is one of the main aspects of ECG forward problem. The simulation results not only are the criterion of the simulation model's precision and reliability, but also have great value in researches and diagnoses. We performed the simulation of QRST waves of complete left bundle branch block (LBBB) and right bundle branch block (RBBB) in virtue of a vector propagation algorithm (VPA), which is accurate, efficient and applicable to anisotropic computer heart models. The simulation results accord with the actual QRST wave in clinical practice.

Algorithms↗

Isotope effects in the photofragmentation of symmetric molecules: the branching ratio of OD/OH in water.

With HOD initially in its vibrational ground state, we present a new detailed interpretation of the OD/OH branching ratio (approximately 3) in the photoinduced process D+OH<--HOD-->H+OD, in the first absorption band. Using semiclassical arguments, we show that the branching ratio has little to do with the initial distribution of configurations, but the initial momentum distribution plays a key role in determination of the branching ratio. The formation of D+OH arises from initial situations where OD is stretching, and it stretches faster than OH, whereas all other motions lead to H+OD. This picture is confirmed by quantum wave-packet calculations.

Journal Article↗

Dynamics of target recognition by interstitial axon branching along developing cortical axons.

Corticospinal axons innervate their midbrain, hindbrain, and spinal targets by extending collateral branches interstitially along their length. To establish that the axon shaft rather than the axonal growth cone is responsible for target recognition in this system, and to characterize the dynamics of interstitial branch formation, we have studied this process in an in vivo-like setting using slice cultures from neonatal mice containing the entire pathway of corticospinal axons. Corticospinal axons labeled with the dye 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (or Dil) were imaged using time-lapse video microscopy of their pathway overlying the basilar pons, their major hindbrain target. The axon shaft millimeters behind the growth cone exhibits several dynamic behaviors, including the de novo formation of varicosities and filopodia-like extensions, and a behavior that we term "pulsation," which is characterized by a variable thickening and thining of short segments of the axon. An individual axon can have multiple sites of branching activity, with many of the branches being transient. These dynamic behaviors occur along the portion of the axon shaft overlying the basilar pons, but not just caudal to it. Once the collaterals extend into the pontine neuropil, they branch further in the neuropil, while the parent axon becomes quiescent. Thus, the branching activity is spatially restricted to specific portions of the axon, as well as temporally restricted to a relatively brief time window. These findings provide definitive evidence that collateral branches form de novo along corticospinal axons and establish that the process of target recognition in this system is a property of the axon shaft rather than the leading growth cone.

Animals↗

Odontoblast processes in dentin revealed by fluorescent Di-I.

There has been controversy about the length and structure of the odontoblast process within dentin since the earliest histologic studies of teeth. Our objective was to use the fluorescent carbocyanine dye Di-I combined with a new gelatin embedment procedure and confocal microscopy to determine the structure and extent of odontoblast processes in developing and mature rat teeth, injured rat molars, reparative dentin, and adult monkey teeth. We found that odontoblast processes do not extend into outer dentin or to the dentin-enamel junction except during early stages of development. Those in innervated regions of crown are long and straight, whereas those in roots are extensively branched and shorter. Cavity injury to crown dentin caused odontoblast fragments to be aspirated into outer dentin. In reparative dentin the odontoblast processes were branched and similar to those in roots. We used photoconversion and electron microscopy to show that Di-I fills the entire odontoblast after gelatin embedment, including the cytoplasm. This is a different type of carbocyanine staining from any previously reported, and it also stains other cells in adjacent hard tissues such as bone and cementum. The Di-I-gelatin method is a new way to use carbocyanine dyes. It has enabled us to solve a long-standing controversy about the histology of teeth, and it should be useful for many other studies of cell structure.

Animals↗

Developmental changes of tau protein and mRNA in cultured rat brain oligodendrocytes.

Oligodendrocytes elaborate an extensive network of multibranched processes and flat membranous sheets. Microtubules (MT) participate in the elaboration and stabilization of myelin-forming processes and are essential for cellular sorting processes. Microtubule-associated proteins (MAPs) are involved in the regulation and stabilization of the dynamic MT network. It has been shown previously that oligodendrocytes express the MAP tau, a phosphoprotein most abundant in neurons of the CNS. In this article, we demonstrate for the first time that oligodendrocytes contain all six tau isoforms, and that tau mRNA and protein expression is developmentally regulated. Immunoblot analysis reveals that tau protein is more abundant, and mature isoforms are more prominent at later stages of development. During the first week of culture maturation, a marked decrease in phosphorylation is observable. Using an RT-PCR approach, we can show that oligodendrocytes express small amounts of exon 3 containing isoforms and that during culture maturation, tau mRNA splice products with 3 MT-binding domains (3R) decrease and mRNA with 4 MT-binding domains (4R) increase. In situ hybridization study demonstrates that tau mRNA is present in precursor cells and in mature oligodendrocytes. Tau mRNA is actively transported into the cellular processes, is specifically present in the primary and some of the secondary processes, enriched at the turning and branching points and the growing tips, and often appears as small patches. Hence, localized tau translation at specific sites in the cellular extensions might contribute to the regulation of MT stability during process formation, early axonal contact establishment, and myelination.

Aging↗

Synthesis of an anionically chargeable, high-molar-mass, second-generation dendronized polymer and the observation of branching by scanning force microscopy.

An efficient synthesis of a methacrylate-based, second-generation (G2) dendronized macromonomer and its free radical polymerization to the corresponding high-molar-mass G2 dendronized polymer are described. The molar mass is determined by gel permeation chromatography (GPC), light-scattering, and analytical ultracentrifugation and compared with values estimated from a scanning force microscopy (SFM) contour lengths analysis of individualized polymer strands on mica. The polymer carries terminal tert-butyl-protected carboxyl groups, the degree of deprotection of which with trifluoroacetic acid is quantified by NMR spectroscopy using the highest molar mass sample. SFM imaging of both protected (noncharged) and unprotected (charged) dendronized polymers on solid substrates reveals mostly linear chains but also some with main-chain branches. The nature of these branches is investigated and the degree roughly estimated to which they are formed. Finally, a synthetic model experiment is described which sheds some light on the aspect of whether chain transfer, a process that could lead to covalent branching, is of importance in the synthesis of the present dendronized polymers.

Journal Article↗

Speciation along environmental gradients.

Traditional discussions of speciation are based on geographical patterns of species ranges. In allopatric speciation, long-term geographical isolation generates reproductively isolated and spatially segregated descendant species. In the absence of geographical barriers, diversification is hindered by gene flow. Yet a growing body of phylogenetic and experimental data suggests that closely related species often occur in sympatry or have adjacent ranges in regions over which environmental changes are gradual and do not prevent gene flow. Theory has identified a variety of evolutionary processes that can result in speciation under sympatric conditions, with some recent advances concentrating on the phenomenon of evolutionary branching. Here we establish a link between geographical patterns and ecological processes of speciation by studying evolutionary branching in spatially structured populations. We show that along an environmental gradient, evolutionary branching can occur much more easily than in non-spatial models. This facilitation is most pronounced for gradients of intermediate slope. Moreover, spatial evolutionary branching readily generates patterns of spatial segregation and abutment between the emerging species. Our results highlight the importance of local processes of adaptive divergence for geographical patterns of speciation, and caution against pitfalls of inferring past speciation processes from present biogeographical patterns.

Animals↗

Computer reconstruction of all the neurons in the optic ganglion of Daphnia magna.

The cellular architecture of the Daphnia compound eye visual system was studied by using computer-aided techniques. All the neurons in one half of the bilaterally symmetric optic ganglion (OG) were reconstructed in three dimensions from serial electron micrographs. The techniques employed were those developed by Levinthal and collaborators (Macagno, Levinthal, and Sobel, Ann. Rev. Biophys. Bioeng. 8:323-351, 1979). The approximately 200 neurons reconstructed were classified according to where they branch in the OG (the lamina and/or the medulla) and whether they send processes to the supraesophageal ganglion and/or across the midplane. Within each class, neurons were further characterized according to cell body location and size and location of their branching fields. Centrifugal processes from neurons with cell bodies not in the OG were also identified. These results provide the bases for a detailed examination of the synaptic connectivity of the identified neurons and for hypotheses concerning their functional roles in visually evoked behaviors.

Animals↗

Bone canaliculus endings in the area of the osteocyte lacuna. Electron-microscopic studies.

Light- and electron-microscopic studies were carried out on the bone canaliculi and their endings in the wall fo the osteocyte lacuna. Two types of canalicular endings were distinguished in the lacuna wall. Those of the first type, fairly numerous, are the endings of small diameter, which do not branch off in the immediate proximity of the lacuna. The other type, occurring in twos or threes, are large in diameter and branch off in the close vicinity of the lacuna, invariably possessing two or three processes which run up to the point of branching of the canaliculus. The images obtained made it possible to demonstrate that the collagen fibrils in both types of endings in the lacuna wall are, in principle, arranged in a similar manner. Nonethless, the type of end formation found where the fibrils run along the axis of the lacuna differs from that for the fibrils in a plexus.

Animals↗

Evaluation of various parts of the paw paw tree, Asimina triloba (Annonaceae), as commercial sources of the pesticidal annonaceous acetogenins.

Various plant parts of the paw paw tree (Asimina triloba Dunal, Annonaceae) were extracted and partitioned to concentrate the mixture of acetogenins into a standardized pesticidal extract (F005). A bioassay with brine shrimp larvae (Artemia salina Leach) was used to determine the relative potencies of the various extracts. The small twigs (0-0.5 cm diameter) yielded the most potent extract (LC50 = 0.04 ppm); the stem wood (LC50 = 4.9 ppm) and leaves (LC50 = 53.7 ppm) yielded the poorest activities. The unripe fruits, seeds, root wood, root bark, and stem bark were notably potent and, generally, yielded > 2% of their dry weight as F005. The smaller diameter stems were more potent than the larger stems. We conclude that, by pollarding the trees, the entire twigs and small branches of paw paw could be processed to produce a potent acetogenin mixture; this biomass could be made available in quantities needed for commercialization of the pesticidal product and could be renewable through regrowth from the parent trunk and larger branches.

Animals↗

Regressive modifications of climbing fibres following Purkinje cell degeneration in the cerebellar cortex of the adult rat.

The role of postsynaptic neurons in the maintenance of adult terminal axon arbours was investigated in the rat olivocerebellar system. The degeneration of Purkinje cells, the main target of olivary axons in the cerebellar cortex, was obtained by intraparenchymal application of kainate. The structural features of target-deprived climbing fibres, visualized by Phaseolus vulgaris leucoagglutinin tracing, were examined from two days to six months after the lesion. Following the degeneration of its Purkinje cell, the climbing fibre underwent remarkable regressive modifications involving the disappearance of most of the terminal arborization. Never the less, atrophic arbours still spanned through the molecular layer six months after the lesion. Morphometric evaluations showed that, one week after kainate application, total arbour length was already reduced to 52% of control, whereas the number of branches and of varicosities had both dropped around 40%. This retraction process progressed in the following stages to reach its maximum at about one month after the lesion, when total length was 30% of control and only 10% of branches and varicosities were still present. Only a slight tendency to a further decrease of the values could be detected at longer survival times. Branching pattern analysis revealed that such regressive phenomena mainly involved the distal compartment of the climbing fibres, the one made of fine varicose branchlets, while sparing the proximal thick branches. In addition, the whole process appeared to follow some rather strict guiding principles leading to an ordered branch retraction, from the periphery of the arbour inwards. Finally, in order to rule out the possibility that the observed changes could be due to a direct action of kainate on climbing fibres, we designed an alternative method of killing Purkinje cells by intraparenchymal injection of propidium iodide. The structural features of climbing fibres deprived of their target by such a procedure were very similar to those shown by arbours from time-matched kainate-lesioned animals at both qualitative and quantitative levels. Our results show that target deprivation induces remarkable structural modifications in the climbing fibre, leading to the retraction of most of the arbour. Never the less, the integrity of the Purkinje cell is not necessary for the maintenance of the whole arborization since its proximal compartment is maintained in the molecular layer for several months after target degeneration. It is proposed that the Purkinje cell, most likely by acting through a contact factor, directly controls the formation and the maintenance of the distal climbing fibre branches with their varicosities, which represent the presynaptic compartment of the axonal arbour.

Animals↗

A Golgi-study of oculomotor neuroblasts migrating across the midline in chick embryos.

The Golgi-Stensaas impregnation technique was employed at appropriate stages of development to study the morphology of the oculomotor neuroblasts as these migrate across the midline. Data reported in previous publications were confirmed, such as the timing of the migration (occurring between the 4th and the 9th days of inoculation), the fact that the migrating cells carry their axons across the midline as trailing processes, and the absence of pre-existing fibrillar structures able to provide contact guidance for this migration. The most striking new fact discovered is that the leading processes of the oculomotor neuroblasts are often branched, and that all these branches are uniformly oriented towards the midline. This seems to indicate the existence of a non-random growth process. It is argued in the discussed that this can be explained through the presence of an orienting neurotropic influence. This one could have its source in certain non-oculomotor neuroblasts which were detected within the midline ventricular zone. On morphological grounds, this group of cells may be tentatively identified as the avian counterpart of the midventral mesencephalic proliferation described in several mammals. This proliferative zone is known to contain dopamine at early stages of development. A hypothetic casual mechanism of the oculomotor migration is therefore advanced, wherein dopamine, diffusing out of the non-oculomotor midline neuroblasts, induces at short range oriented outgrowth of oculomotor leading processes across the midline.

Animals↗

Zero-range model of traffic flow.

A multicluster model of traffic flow is studied, in which the motion of cars is described by a stochastic master equation. Assuming that the escape rate from a cluster depends only on the cluster size, the dynamics of the model is directly mapped to the mathematically well-studied zero-range process. Knowledge of the asymptotic behavior of the transition rates for large clusters allows us to apply an established criterion for phase separation in one-dimensional driven systems. The distribution over cluster sizes in our zero-range model is given by a one-step master equation in one dimension. It provides an approximate mean-field dynamics, which, however, leads to the exact stationary state. Based on this equation, we have calculated the critical density at which phase separation takes place. We have shown that within a certain range of densities above the critical value a metastable homogeneous state exists before coarsening sets in. Within this approach we have estimated the critical cluster size and the mean nucleation time for a condensate in a large system. The metastablity in the zero-range process is reflected in a metastable branch of the fundamental flux-density diagram of traffic flow. Our work thus provides a possible analytical description of traffic jam formation as well as important insight into condensation in the zero-range process.

Journal Article↗

[Virtual bronchoscope system].

In this article, we describe the features of Virtual Bronchoscope System(VBS) and its practical use. VBS is constructed based on 3-D chest CT images. The bronchus region is automatically extracted from 3-D chest CT images by a three-dimensional region growing method. The surface rendering is employed for construction of virtualized tracheo-bronchial tree. It gives us an environment where we can observe inside the bronchi from an arbitrary viewpoint and a view direction. By mouse operation, the user can control the viewpoint and the view direction to fly through inside the airway in real time. VBS is applicable for a variety of purposes such as diagnosis, surgical planning, informed consent, education and training. One of extension of this system is a teaching tool for medical students. In the module for educational use, we have developed four functions for using the system as a teaching tool as follows: (a) automated display of bronchial anatomical names, (b) presenting questions about the currently observed branch in the endoscopic view, (c) display of the path which the user should follow, and (d) display of a question about the location of the artificially created tumor in the bronchus. These functions use the processed results of automated anatomical labeling. The method proposed here combines the knowledge based processing technique 'automated labeling of bronchial branch' and the novel visualization technique 'virtual bronchoscope'. This is one of new teaching tools of medical images. We conclude that this virtual bronchoscope system might have an important role in the medial students' education.

Bronchi↗

Interactions between segmental homologs and between isoneuronal branches guide the formation of sensory terminal fields.

Process outgrowth and peripheral field innervation by an identified mechanosensory neuron were examined in the intact embryonic leech. The dorsal pressure-sensitive (PD) neurons of the leech CNS are found as bilateral pairs in every segmental ganglion, and are amenable to study at early ages in intact embryos. Each PD has one major axonal projection that putatively pioneers the nerve to the dorsal body wall and branches extensively in its own segment, and two minor projections that innervate, via neighboring ganglia, smaller areas in adjacent segments. We found that adjacent embryonic PD cells form overlapping terminal fields in the body wall, but that the extent of overlap was governed by inhibitory interactions among these fields. When one PD neuron was ablated, the adjacent PD cell changed its peripheral arborization by (1) its major axon producing more filopodia and extending longer side branches toward the ablated cell and (2) its minor axon producing a large arbor in the operated segment. Interestingly, although growth was biased toward the side of the ablated neuron, reduced outgrowth of the PD cell was found on the side away from the ablation, while the total extent of arborization of the PD cell kept relatively constant. Further, we found that axotomy of the major PD projection resulted in extensive outgrowth of its minor projections. These results suggest that a single PD neuron has a limited capacity for growth, each of its branches growing at the expense of the others, and that inhibitory interactions between neighboring PD neurons influence the extent and direction of that growth.

Animals↗

Preference for a solution of branched-chain amino acids plus glutamine and arginine correlates with free running activity in rats: involvement of serotonergic-dependent processes of lateral hypothalamus.

Rats were given free access to a running wheel, food, water, and a solution composed of branched-chain amino acids plus glutamine and arginine (the "BCAA-based" solution). A positive relationship between dark-period running distance and preference for the BCAA-based solution was observed. Serotonin release in the lateral hypothalamus, the central nucleus of amygdala and the medial nucleus raphe in overnight fluid-deprived rats during their first subsequent free drinking was also measured. A lowered serotonin release in the lateral hypothalamus characterized the rats that consumed the BCAA-based solution. No drink-related changes were observed in the amygdala. A separate group of rats was trained on a treadmill. Following the training period, plasma amino acids and brain serotonin release were measured during running. The BCAA-based solution infused before running elevated the branched chain amino acids/tryptophan plasma ratio at the end of, and after, running. Additionally, a decreased lateral hypothalamus serotonin release was seen 80 min after running, when compared with water-infused rats. No fluid-related changes in the amygdala were observed. The exercise-related shift in the fluid preference towards a BCAA-based solution suggests an ergogenic benefit. The forced-running study shows the lateral hypothalamus as a critical region in the effects of a BCAA-based solution.

Amino Acids, Branched-Chain↗

[Postreplication DNA repair in Escherichia coli cells. I. Defect of inducible repair in the thermosensitive mutant AX727 dnaZts at a nonpermissive temperature].

After UV-irradiation at the non-permissive temperature 43 degrees C the survival of the thermosensitive mutant Escherichia coli AX727 dnaZts (the dnaZ gene controls the synthesis of gamma-subunit of DNA polymerase III holoenzyme) appeared somewhat lower and postreplication perair of its DNA less effective by 5-15% than at the permissive temperature 30 degrees. The survival of chloramphenicol treated cells decreased both at 43 and 30 degrees C. Under the influence of chloramphenicol, the postreplication repair of DNA in AX727 dnaZts cells at 30 degrees lower to the level of this process seen at 43 degrees C. The same process in thermoresistant revertant AX729 dnaZtr at 30 degrees and 43 degrees C decreased to the level of the process in AX727 cells at 43 degrees C. The data indicate that dnaZ gene product is necessary for the inducible branch of postreplication DNA repair. By this process (presumably together with prereplication excision repair), the cell eliminates 20% of UV-inducible lethal damages. Besides, in AX727 dnaZts cells there functions protein synthesis dependent repair not associated with the postreplication repair of DNA; this latter process eliminates about 60% lethal damages of the cell.

DNA Repair↗